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Sample records for kinetin

  1. Kinetin Reversal of NaCl Effects

    Katz, Adriana; Dehan, Klara; Itai, Chanan

    1978-01-01

    Leaf discs of Nicotiana rustica L. were floated on NaCl in the presence of kinetin or abscisic acid. On the 5th day 14CO2 fixation, [3H]leucine incorporation, stomatal conductance, and chlorophyll content were determined. Kinetin either partially or completely reversed the inhibitory effects of NaCl while ABA had no effect. PMID:16660618

  2. Kinetin (N -furfuryladenine): Cytotoxicity against MCF-7 breast ...

    Jane

    2011-07-06

    Jul 6, 2011 ... The cytotoxicity effect of kinetin on MCF-7 breast cancer cell lines was ... Medium (DMEM) containing 10% FBS, 2 mM glutamine, 100 units/ml ..... apoptosis of human myeloid leukemia cells by cytokinins and cytokinin ...

  3. Aplication Of PGR IAA And Kinetin On Rubber Clones To Accelarate Mature Tapping

    Try Koryati

    2017-09-01

    Full Text Available This study aims to determine the most appropriate concentration of IAA hormones Kinetin on rubber clones to accelerate the mature tapping. Experiments have been carried out in Karang Inong Plantation PTP N I Langsa East Aceh. This study is arranged in two factors of Nested Design. Clones factor is consisted of 5 level and IAA hormon Kinetin factor has 7 levels. The results showed that the concentration of IAA hormones Kinetin is different to each clone to accelerate mature tapping. Application of IAA hormone Kinetin significantly affect the parameters of girth bark thickness number of latex vessels latex vessel diameter leaf area and the amount of chlorophyll. Application of IAA 600ppm Kinetin 60 ppm H6 show the largest increase on the girth. Clone treatment is also has significant effect on all parameters but the largest girth found in clones PB 330 and IRR 5. Combination of Clones with IAA hormone Kinetin significantly affects the bark thickness the number of latex vessels and latex vessel diameter. Application of IAA 400 ppm Kinetin 60 ppm has no significant effect on girth but able to accelerate the mature tapping particularly for clones PB 330 and IRR5 K2H4 and K5H4 as indicated by girth size namely 48.15 cm and 48.20 cm respectively at planting age 46 months.

  4. The effect of kinetin on wheat seedlings exposed to boron.

    Eser, Ahmet; Aydemir, Tülin

    2016-11-01

    The objective of this study was to examine relationship between boron (B) induced oxidative stress and antioxidant system in boron sensitive and tolerant wheat cultivars Bezostaya and Kutluk, and also to investigate whether Kinetin (KN) enhances the level of antioxidant system, relative growth, concentration of hydrogen peroxide (H 2 O 2 ), malondialdehyde (MDA) and proline and chlorophyll content in both cultivars exposed to B stress. B treatments diminished growth and chlorophyll content whereas, it enhanced accumulation of H 2 O 2 , MDA and proline, and various antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and lipoxygenase (LOX) in the shoot and root of both cultivars. However, the follow-up application of KN to the B stressed plants improved growth and chlorophyll content and further enhanced the mentioned antioxidant enzymes and level of H 2 O 2 , MDA and proline. This study thus suggests that KN improves B tolerance of the studied cultivars grown under B toxicity. Copyright © 2016. Published by Elsevier Masson SAS.

  5. Kinetin improves IKBKAP mRNA splicing in patients with familial dysautonomia

    Axelrod, Felicia B.; Liebes, Leonard; Gold-von Simson, Gabrielle; Mendoza, Sandra; Mull, James; Leyne, Maire; Norcliffe-Kaufmann, Lucy; Kaufmann, Horacio; Slaugenhaupt, Susan A.

    2011-01-01

    Familial dysautonomia (FD) is caused by an intronic splice mutation in the IKBKAP gene that leads to partial skipping of exon 20 and tissue-specific reduction in I-κ-B kinase complex associated protein/ elongation protein 1 (IKAP/ELP-1) expression. Kinetin (6-furfurylaminopurine) has been shown to improve splicing and increase wild-type IKBKAP mRNA and IKAP protein expression in FD cell lines and carriers. To determine if oral kinetin treatment could alter mRNA splicing in FD subjects and was tolerable, we administered kinetin to eight FD individuals homozygous for the splice mutation. Subjects received 23.5 mg/Kg/day for 28 days. An increase in wild-type IKBKAP mRNA expression in leukocytes was noted after eight days in six of eight individuals; after 28 days the mean increase as compared to baseline was significant (p=0.002). We have demonstrated that kinetin is tolerable in this medically fragile population. Not only did kinetin produce the desired effect on splicing in FD patients, but also that effect appears to improve with time despite lack of dose change. This is the first report of a drug that produces in vivo mRNA splicing changes in individuals with FD and supports future long-term trials to determine if kinetin will prove therapeutic in FD patients. PMID:21775922

  6. The influence of gibberellic acid and kinetin on the growth of Scenedesmus quadricauda (Turp. Breb.

    J. Buczek

    2015-01-01

    Full Text Available The influence of gibberellic acid (GA3 and of kinetin (6-furfurylamino purine on the increment of cell number increase in dry weight and upon protein level in Scenedesmus quadricauda (Turp. Bréb. was studied. It was found that 10-7 M GA3 stimulates at the same time cell growth and dry weight increase of the algae. No influence of GA3 upon the protein content was observed. Kinetin of 10- M concentration stimulates in the initial growth phase cell multiplication and increases the protein level. This substance promotes the increment in dry weight however in the later phase of growth. Furthermore kinetin prolongs the viability of algae, extending the growth phase.

  7. Combined effect of kinetin and radiation treatment on the cap opening of Agaricus bisporus

    Kovacs, E.

    1982-01-01

    Cultivated mushrooms (Agaricus bisporus) with a cap diameter of 3-5 cm were incubated at 20 deg C during the experimental period (30-72 h). The degree of cap opening was determined and the data were evaluated. It was found that kinetin solutions in the 0 to 32 mg kg -1 range stimulated the opening of the mushroom cap. The shorter the time elapsed between picking and treating the mushrooms, the greater was the effect. A kinetin solution with a concentration of 100 mg kg -1 inhibited the opening of the cap. Cap opening in irradiated mushrooms cannot be induced even with kinetin concentrations that stimulate cap opening. Treatment with 0, 2.5 and 10 kGy doses of radiation the stimulating effect on cap opening decreased as a function of the rising dose. The radiation effects in kinetin solutions and various natural cytokinins (zeatin, 2iP and 2iPA) were studied and it was found that cytokinin solutions lost 50-60% of their activity after a radiation dose of only 1 kGy. (author)

  8. Mechanistic investigation on the oxidation of kinetin by Ag(III ...

    Administrator

    aging effects in human skin cells and other systems. At present, kinetin is one of ... The stability of Ag(OH). –. 4 is very ... pore water was used throughout the work. A solution .... The reaction orders were determined from the slope of log kobs vs ...

  9. Mechanistic investigation on the oxidation of kinetin by Ag(III)

    Home; Journals; Journal of Chemical Sciences; Volume 122; Issue 6. Mechanistic investigation on the oxidation of kinetin by Ag(III) periodate complex in aqueous alkaline media: A kinetic approach. S D Lamani A M Tatagar S T Nandibewoor. Full Papers Volume 122 Issue 6 November 2010 pp 891-900 ...

  10. Effects of putrescine, kinetin and IAA on protein synthesis in 'Phaseolus vulgaris' coleoptiles

    Crocomo, O J; Lee, T S.G. [Centro de Energia Nuclear na Agricultura, Piracicaba (Brazil)

    1975-01-01

    Incubation of etiolated 'Phaseolus vulgaris' coleoptiles shows a converse flux between soluble protein and reducing sugar. The rate of incorporation of radioactive arginine into protein was higher than that of radioactive leucine. Radioactive arginine incorporation into protein was linear up to 120 min and then started to decline. The rate of incorporation of radioactive leucine was increased by preincubation of the tissue in the incubation medium. Roots were found to contain more soluble protein and much less reducing sugar than the coleoptile. The optimum pH value for protein synthesis in coleoptile sections was found to be 6 for control tissues and 4 for those treated with 10-/sup 3/M IAA. This high concentration of IAA was also found to inhibit soluble protein synthesis, the incorporation rate of radioactive arginine and leucine into protein fraction, the secretion of hydrogen ion into the incubation medium and elongation of the bean segment. Kinetin at 2x10/sup -4/M and putrescine at 5mM both decreased the rate of /sup 14/C-arginine incorporation into soluble protein, but for /sup 14/C-leucine, this rate of incorporation was found to be increased after 90 min incubation with a preincubation of 30 min. In general, the change pattern of the soluble protein content, the reducing sugar level and the incorporation rate of radioactive arginine and leucine into protein in the kinetin and putrescine treated tissues were about the same although tissues that incubated with kinetin always contain more soluble protein and less reducing sugar than that of incubated with putrescine.

  11. Physiological response of marigold (calendula officinalis L.) plants to gamma radiation, gibberellic acid and kinetin

    Noby, M.F.A.

    2010-01-01

    This study was carried out during the two successive seasons of 2005/2006 and 2006/2007 at the Experimental Field of Plant Research Department, Nuclear Research Center, Atomic Energy Authority at Inshas in a newly reclaimed sandy loam soil. The aim of this work was to study the effect of gamma radiation, gibberellic acid or kinetin and their interaction on the growth, flowering and the productivity of pot-marigold (Calendula officinalis L.) plants. The experimental trials included two factorial experiments; the first one was to study the effect of gamma radiation and gibberellic acid and the interaction between them on pot-marigold plants. Another experiment was conducted to study the effect of gamma radiation and kinetin and the interaction between them on pot-marigold plants. Pot-marigold seeds were irradiated before sowing with gamma rays at 0, 50, 100, 150 or 200 Gray (Gy) of gamma rays. After planting, plants were sprayed with either gibberellic acid (at the concentrations of 0, 50, 100, 150 or 200 ppm) or kinetin (at the concentrations of 0, 10, 20, 30 or 40 ppm). Generally, gamma rays treatments (50, 100 and 150 Gy) increased plant height, branch number/plant, leaf area, vegetative growth and roots fresh and dry weights of pot-marigold plants. Also, the same gamma doses accelerated flowering and decreased the period from sowing until flowering while increased flower head diameter, flower number/plant and flowers fresh and dry weights per plant and per feddan. In addition, gamma rays (50 - 150 Gy) increased volatile oil yield in flowers, leaf chlorophyll content, carotenoids and beta carotene in flowers, total soluble sugars and NPK %. The best values were obtained by 50 Gy dose of gamma rays, whereas the dose of 200 Gy gave the lowest values.

  12. Water and Salt Stresses, Kinetin and Protein Synthesis in Tobacco Leaves 1

    Ben-Zioni, Aliza; Itai, C.; Vaadia, Y.

    1967-01-01

    The capacity of tobacco (Nicotiana rustica) leaf discs to incorporate l-leucine 14C into proteins was measured. Leaf discs were obtained from plants which experienced soil water depletion, or which were exposed to a saline or osmotic stress in the root medium. The stresses were brief of relatively short duration and water potential did not decrease below 4 bars in the root media. Leaf discs were sampled 2 hours after stress removal, achieved by reirrigation, or replacement of saline and osmotic solutions with normal nutrient solution. Plants were always turgid when leaves were sampled. All stressed tissues showed reduced capacity to incorporate l-leucine 14C into protein. The reduction was about 50% and could not be attributed either to reduced uptake into the discs, or to possible isotopic dilution. Incorporation decreased progressively with leaf age in control discs as well as in stressed leaf discs. At all ages tested, incorporation in stressed discs was lower than that of the control. Full recovery of incorporation capacity in stressed discs was obtained when discs were sampled 72 hours after stress removal but not earlier. Kinetin pretreatment prior to incubation with labelled leucine partially restored incorporation in stressed discs. The differences in response to kinetin of stressed and control discs suggest a lower endogenous level of cytokinins in the stressed discs. The results were qualitatively similar regardless of the kind of stress given to the plants during pretreatment. This supports the hypothesis that the normal supply of root cytokinins is important in shoot metabolism. PMID:16656515

  13. Physiological and Fluorescence Reaction of Four Rice Genotypes to Exogenous Application of IAA and Kinetin under Drought Stress

    Mostafa SALEHIFAR

    2017-09-01

    Full Text Available To assess the effects of IAA and Kinetin plant growth regulators in order to improve the drought tolerance in rice seedlings (Oryza sativa L., a factorial experiment was carried out based on complete randomized design with three replications. The experimental factors included different rice genotypes [‘Gharib’, ‘Khazar’, ‘Sepidrood’ and ‘IR83750 -131-1’ (‘IR83750’ ], drought stress from 1 to 4 code of the Vergara coding system and control (normal irrigation and growth regulators in three levels (IAA and Kinetin through foliar spraying and non-application as control. The results indicated, under normal irrigation condition together with IAA application, ‘IR83750’ rice had the highest number of tillers and leaf greenness, with mean of 18.27 and 49.46, respectively. The highest amount of leaf relative water content 95.11 percent was related to ‘Sepidrood’. Under drought stress condition, the highest electrolyte leakage (36.59 percent was observed in ‘Gharib’. In drought condition, the highest leaf drying score was related to ‘Gharib’ in both years, but the highest score of leaf rolling index (9 was observed in ‘Gharib’ and ‘Khazar’. The present findings showed that drought stress had harmful effects in all examined genotypes and the impact in susceptible genotypes (‘Gharib’ and ‘Khazar’ was more than ‘IR83750’ and ‘Sepidrood’. Application of growth regulators (IAA and Kin improved conditions for the growth of all genotypes. Therefore, using the tolerant genotypes along with growth regulators can improve the rice growth traits.

  14. Effect of gibberellin, auxin and kinetin treatments combined with foliar applied NPK on the yield of Capsicum annuum L. fruits and their capsaicin content

    Tomasz J. Nowak

    2013-12-01

    Full Text Available The Wrocław version of hydroponic culture was applied. Under optimal conditions of root fertilization the plants were sprayed with growth regulators sueh as gibberellins, auxins and kinetins, and .their mixtures. Each growth regulator treatment was applied with or without NPK added. The influence of these treatments on the fresh and dry weight of the fruit, percentage of ripe fruits and content and yield of capsaicin was studied. The highest yield of fruits and capsaicin. was obtained from plants sprayed with gibberellic acid and kinetin (in concentrations of 10 and 5 mg/l, respectively together with NPK foliar application. No influence of ,growth regulators and foliar-applied NPK was noted on capsaicin content and dry weight of fruits.

  15. Vitamin C and total phenols quantification in calli of native passion fruit induced by combinations of Picloram and Kinetin

    Fabiane Aparecida Artioli-Coelho

    2015-08-01

    Full Text Available Brazil is one of the center of origin of passion fruit and has an important natural variability of the genus Passiflora. Several wild species of this genus are resistant to some pests and diseases and many are considered as medicinal. The aim of this research was to induce callus from in vitro Passiflora gibertii leaf explants for quantification of vitamin C and total phenols. Once the appropriate auxin/cytokine balance promotes callus formation and may optimize the production of secondary compounds and vitamins, calli were induced using a half-strength MS medium with a combination of the auxin Picloran (PIC and the cytokine Kinetin (KIN. The vitamin C and total phenols were quantified by colorimetric methods from calli after different culture periods. The calli induction was strongly dependent of the combination PIC/KIN. It was observed high vitamin C content (94.8mg 100g-1 during the calli induction period in MS+4.14µM PIC+ 0.207µM KIN. Higher PIC/KIN concentrations promoted an increase in the vitamin C content after three subcultures. The higher PIC (8.28µM/KIN (0.828µM concentration was the higher was the total phenols production (66mg tannic acid 100g-1 of fresh callus during the calli induction period.

  16. Kinetin Enhanced 1-Aminocyclopropane-1-Carboxylic Acid Utilization during Alleviation of High Temperatures Stress in Lettuce Seeds.

    Khan, A A; Prusinski, J

    1989-10-01

    The thermoinhibition at 35 and 32 degrees C of pregermination ethylene production and germination in lettuce (Lactuca sativa L. cv Mesa 659) seeds was synergistically or additively alleviated by 0.05 millimolar kinetin (KIN) and 10 millimolar 1-aminocyclopropane-1-carboxylic acid (ACC). The synergistic effect of KIN + ACC on ethylene production and germination at 35 degrees C was inhibited by Co(2+) (44-46%) but not by aminoethoxyvinyl glycine (AVG). The uptake of ACC by the seed was not influenced by KIN. Upon slitting of the seed coats (composed of pericarp, testa and endosperm), following the uptake of chemicals, ACC was readily converted into ethylene at all temperatures, and the synergistic effects of KIN + ACC at 35 degrees C were lost. At 35 degrees C, KIN acted synergistically with ACC or ethephon (ETH) in alleviating the osmotic restraint. At 25 degrees C, ETH was more active than KIN or KIN + ACC in overcoming the osmotic restraint. Thus, the integrity of the seed coats, the KIN-enhanced ACC utilization, and an interaction of KIN with the ethylene produced may be the basis for the synergistic or additive effects of KIN + ACC at high temperature.

  17. Callus induction of leaf explant Piper betle L. Var Nigra with combination of plant growth regulators indole-3-acetic acid (IAA), benzyl amino purin (BAP) and kinetin

    Junairiah, Zuraidassanaaz, Nabilah Istighfari; Izdihar, Fairuz Nabil; Manuhara, Yosephine Sri Wulan

    2017-09-01

    The purpose of this research was to determine the combination of plant growth regulators IAA, BAP and kinetin towards callus induction and growth of leaf explants Piper betle L. VarNigra. Explants from leaf of Piper betle L. VarNigra was cultured on MS medium with 24 treatment combinations of plant growth regulators IAA and BAP and 24 treatment combinations of plant growth regulators IAA and kinetin with 0.0;0.5;1.0;1.5;2.0 mg/L concentration respectively, the observed variable were the length of time the formation of callus, callus morphology, fresh and dry weight of callus. The results of this research showed that the combination of growth regulators IAA with BAP and kinetin had effects on leaf growth of Piper betle L. VarNigra. During 8 weeks observation, it indicated that the combination of concentration IAA 0.5 mg/L and BAP 2.0 mg/L showed fastest callus formation at 8.5 days. Combination of concentration IAA 1.0 mg/L and BAP 1.5 mg/L showed the highest of fresh weight at 0.6596 grams, and the highest dry weight was obtained from the combination of concentration IAA 0.5 mg/L and BAP 0.5 mg/L at 0.0727 grams. Combination of concentration IAA 1.0 mg/L and kinetin 1.5 mg/L had the highest of fresh weight at 0.2972 grams and the highest dry weight at 0.1660 grams. Callus of Piper betle L. VarNigra had two textures, that were compact and friable, and also showed various kind of colors, like white, greenish white, yellowish white, tanned white, brown and black. Based on this research, that concentration IAA 1.0 mg/L and 1.5 mg/L kinetin was the best combination for induction of callus from leaf of Piper betle L. Var Nigra.

  18. Comparative Study for the Effect of Gibberellic acid, Kinetin and Indole-3-acetic acid on Seed Germination performance of Dianthus caryophyllus

    Rajib Roychowdhury

    2012-07-01

    Full Text Available Seed germination is the major limiting factor for large-scale production and cultivation of crop species. Such attribute also positively as well as negatively affected by some potent plant growth regulators and other chemical compounds. For this, present experiment was undertaken with an objective to investigate the comparison of the effect of various concentrations of plant growth regulators like Gibberellic Acid (GA3, Kinetin and Indole-3-acetic acid (IAA on seed germination of Dianthus caryophyllus or Carnation. Dianthus seeds were soaked in different concentrations (0 ppm or control, 10, 20, 30 and 40 ppm of each of GA3, Kinetin and IAA for 24 h at room temperature (25 ± 2°C. Three replicates of each treatment with ten seeds per replicate were arranged for precise physiological analysis. Significant variation was found in all aspects after analysis of variance (ANOVA of each mean value. After two weeks of seed soaking, it was noted that germination percentages were significantly accelerated by lower concentrations (10 and 20 ppm of used hormones. Amongst the three potential growth regulators, 20 ppm was found most effective because it showed highest germination percentage for GA3 (87.46%, Kinetin (78.92% and IAA (75.35%. A great deal of information relating to seed germination practices shows that these plant growth regulators were efficient to overcoming dormancy leading to rapid seed germination. GA3 was selected as best hormone, in this study, which showed highest seed germination. These results could useful to large-scale cultivation of Dianthus caryophyllus plants for improving its floricultural impact worldwide.

  19. Morphological aspects of starch and cell wall material mobilization in developing lupine cotyledons and the effect of kinetin on these processes

    Fortunat Młodzianowski

    2015-01-01

    Full Text Available In the cotyledons of dry lupine seeds the presence of starch was not demonstrated. Its formation during seed imbibition in darkness is accompanied by a reduction in the thickness of cell walls containing hemicelluloses. It is believed that the products of hemicellulose hydrolysis, particullarily in isolated cotyledons, arę the main source of materials for the synthesis of starch, In the process of cell wall decomposition the invaginations of plasmalemma appear to be involved. Kinetin enhance the hydrolysis of cell walls and the mobilization of starch in isolated cotyledons.

  20. Translocation of labelled assimilates, ion uptake and nucleic acids contents in zea mays plants as influenced by application of the herbicide dual and the bioregulaators GA3 and kinetin

    Hassanein, R.AA.; Khodary, S.E.A.; Abdel-Aziz, S.M.

    2001-01-01

    Maize seedlings, grown hydroponic for one month, were undertaken o investigate the effect of dual (metolachlor), bio regulators (GA 3 and kinetin) and their interaction with dual on translocation rate of assimilates, nucleic acids content. ion uptake and the activities of protease and nitrate reductase enzymes. Dual at all concentrations decreased the rate of assimilates translocation and nucleic acids levels. Also reduction in the ability of the treated plants to absorb ions from the growth medium as well as the activities of nitrate reductase and protease enzymes were retarded upon dual application. The results also revealed that treatment with either GA 3 or kinetin in combination with dual, reversed the adverse action of the herbicide on zea mays plants

  1. FORMULASI AUKSIN (INDOLE ACETIC ACID DAN SITOKININ (KINETIN, ZEATIN UNTUK MORFOGENESIS SERTA PENGARUHNYA TERHADAP PERTUMBUHAN, SINTASAN DAN LAJU REGENERASI KALUS RUMPUT LAUT, Kappaphycus alvarezii

    Sri Redjeki Hesti Mulyaningrum

    2013-03-01

    Full Text Available Interaksi auksin dan sitokinin dianggap penting untuk mengatur pertumbuhan dan perkembangan dalam kultur jaringan tanaman. Penelitian ini bertujuan untuk menentukan komposisi auksin dan sitokinin yang optimum untuk morfogenesis kalus rumput laut K. alvarezii, dan mengevaluasi pengaruhnya terhadap pertumbuhan, sintasan, dan laju regenerasi kalus. Kultur kalus dilakukan pada media cair dengan formulasi zat pengatur tumbuh (ZPT indole acetic acid (IAA : kinetin : zeatin, dengan komposisi konsentrasi sebagai berikut: A 0,4:0:1 mg/L; B 0,4:0,25:0,75 mg/L; C 0,4:0,5:0,5 mg/L; D 0,4:0,75: 0,25 mg/L; E 0,4:1:0 mg/L; kontrol (tanpa ZPT. Desain penelitian adalah rancangan acak lengkap dengan pengulangan tiga kali untuk masing-masing perlakuan. Parameter yang diamati adalah laju pertumbuhan harian, sintasan, laju regenerasi, panjang tunas, dan morfologi tunas. Analisis data dilakukan dengan uji keragaman (ANOVA dan hasil yang diperoleh disajikan dalam bentuk grafik. Hasil penelitian menunjukkan bahwa formula optimum untuk morfogenesis rumput laut K. alvarezii adalah formula A dengan komposisi IAA : zeatin = 0,4:1 mg/L. Penggunaan formula zat pengatur tumbuh yang berbeda berpengaruh nyata (P0,05 terhadap sintasan kalus. Tunas rumput laut K. alvarezii mulai terbentuk pada hari ke-15 masa kultur.

  2. Improvement in shelf life of minimally processed cilantro leaves through integration of kinetin pretreatment and packaging interventions: Studies on microbial population dynamics, biochemical characteristics and flavour retention.

    Ranjitha, K; Shivashankara, K S; Sudhakar Rao, D V; Oberoi, Harinder Singh; Roy, T K; Bharathamma, H

    2017-04-15

    Effect of integrating optimized combination of pretreatment with packaging on shelf life of minimally processed cilantro leaves (MPCL) was appraised through analysis of their sensory attributes, biochemical characteristics, microbial population and flavour profile during storage. Minimally pretreated cilantro leaves pretreated with 50ppm kinetin and packed in 25μ polypropylene bags showed a shelf life of 21days. Optimized combination helped in efficiently maintaining sensory parameters, flavour profile, and retention of antioxidants in MPCL until 21days. Studies conducted on the effect of optimized combination on microbial population and flavour profile revealed that among different microorganisms, pectinolysers had a significant effect on spoilage of MPCL and their population of ⩽3.59logcfu/g was found to be acceptable. Principal component analysis of headspace volatiles revealed that (E)-2-undecenal, (E)-2-hexadecenal, (E)-2-tetradecenal & (E)-2-tetradecen-1-ol in stored samples clustered with fresh samples and therefore, could be considered as freshness indicators for MPCL. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Fate of nuclear material during subsequent steps of the kinetin-induced PCD in apical parts of Vicia faba ssp. minor seedling roots.

    Kaźmierczak, Andrzej; Soboska, Kamila

    2018-07-01

    In animals during apoptosis, the best examined type of programmed cell death (PCD), three main phases are distinguished: (i) specification (signaling), (ii) killing and (iii) execution one. It has bean postulated that plant PCD also involves three subsequent phases: (i) transmission of death signals to cells (signaling), (ii) initiation of killing processes and (iii) destruction of cells. One of the most important hallmarks of animal and plant PCD are those regarding nucleus, not thoroughly studied in plants so far. To study kinetin-induced PCD (Kin-PCD) in the context of nuclear material faith, 2-cm apical parts of Vicia faba ssp. minor seedling roots were used. Applied assays involving spectrophotometry, transmission electron microscopy, fluorescence and white light microscopy allowed to examine metabolic and cytomorphologic hallmarks such as changes in DNA content, ssDNA formation and activity of acidic and basic nucleases (DNases and RNases) as well as malformations and fragmentation of nucleoli and nuclei. The obtained results concerning the PCD hallmarks and influence of ZnSO 4 on Kin-PCD allowed us to confirmed presence of specification/signaling, killing and execution/degradation phases of the process and broaden the knowledge about processes affecting nuclei during PCD. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. Synergistic enhancement of ethylene production and germination with kinetin and 1-aminocyclopropane-1-carboxylic Acid in lettuce seeds exposed to salinity stress.

    Khan, A A; Huang, X L

    1988-08-01

    Relief of salt (0.1 molar NaCl) stress on germination of lettuce (Lactuca sativa L., cv Mesa 659) seeds occurred with applications of 0.05 millimolar kinetin (KIN) and 1 to 10 millimolar 1-aminocyclopropane 1-carboxylic acid (ACC). Treatment with KIN enhanced the pregermination ethylene production under saline condition. A synergistic or an additive enhancement of pregermination ethylene production and germination occurred under saline condition in the presence of KIN and a saturating dose (10 millimolar) of ACC. No KIN-ACC synergism was noted in ethylene production or germination under nonsaline condition. Addition of 1 millimolar aminoethoxyvinylglycine (AVG) inhibited the KIN-enhanced pregermination ethylene production (85 to 89%) and germination (58%) under saline condition but not the synergistic effect of KIN + ACC on ethylene production. Under nonsaline condition, AVG had no effect on germination even though ethylene production was strongly inhibited. Alleviation of salt stress by KIN was inhibited in a competitive manner by 2,5-norbornadiene (NBD) (0.02-0.2 milliliter per liter), and the addition of ACC and/or ethylene reduced this inhibition. An increase in the pregermination ethylene production and germination occurred also by cotylenin E (CN) under saline condition. However, neither AVG (1 millimolar) nor NBD (0.02 to 0.2 milliliter per liter) prevented the relief of salt stress by CN. Thus, KIN may alleviate salt stress on germination by promoting both ACC production and its conversion to ethylene. Rapid utilization of ACC may be the basis for the synergistic or the additive effect of KIN plus ACC. The need for ethylene production and action for the relief of salt stress is circumvented by a treatment with CN.

  5. Requirement for ethylene synthesis and action during relief of thermoinhibition of lettuce seed germination by combinations of gibberellic acid, kinetin, and carbon dioxide

    Saini, H.S.; Consolacion, E.D.; Bassi, P.K.; Spencer, M.S.

    1986-01-01

    Application of exogenous ethylene in combination with gibberellic acid (GA 3 ), kinetin (KIN), and/or CO 2 has been reported to induce germination of lettuce seeds at supraoptimal temperatures. However, it is not clear whether endogenous ethylene also plays a mediatory role when germination under these conditions is induced by treatment regimes that do not include ethylene. Therefore, possible involvement of endogenous ethylene during the relief of thermoinhibition of lettuce (Lactuca sativa L. cv Grand Rapids) seed germination at 32°C was investigated. Combinations of GA 3 (0.5 millimolar), KIN (0.05 millimolar), and CO 2 (10%) were used to induce germination. Little germination occurred in controls or upon treatment with ethylene, KIN, or CO 2 . Neither KIN nor CO 2 affected the rate of ethylene production by seeds. Both germination and ethylene production were slightly promoted by GA 3 . Treatments with GA 3 + CO 2 , GA 3 + KIN, or GA 3 + CO 2 + KIN resulted in approximately 10- to 40-fold increases in ethylene production and 50 to 100% promotion of germination as compared to controls. Initial ethylene evolution from the treated seeds was greater than from the controls and a major surge in ethylene evolution occurred at the time of visible germination. Application of 1 millimolar 2-aminoethoxyvinyl glycine (AVG), an inhibitor of ethylene synthesis, in combination with any of above three treatments inhibited the ethylene production to below control levels. This was accompanied by a marked decline in germination percentage. Germination was also inhibited by 2,5-norbornadiene (0.25-2 milliliters per liter), a competitive inhibitor of ethylene action. Application of exogenous ethylene (1-100 microliters per liter) overcame the inhibitory effects of AVG and 2,5-norbornadiene on germination. The results demonstrate that endogenous ethylene synthesis and action are essential for the alleviation of thermoinhibition of lettuce seeds by combinations of GA 3 , KIN, and CO

  6. Requirement for Ethylene Synthesis and Action during Relief of Thermoinhibition of Lettuce Seed Germination by Combinations of Gibberellic Acid, Kinetin, and Carbon Dioxide.

    Saini, H S; Consolacion, E D; Bassi, P K; Spencer, M S

    1986-08-01

    Application of exogenous ethylene in combination with gibberellic acid (GA(3)), kinetin (KIN), and/or CO(2) has been reported to induce germination of lettuce seeds at supraoptimal temperatures. However, it is not clear whether endogenous ethylene also plays a mediatory role when germination under these conditions is induced by treatment regimes that do not include ethylene. Therefore, possible involvement of endogenous ethylene during the relief of thermoinhibition of lettuce (Lactuca sativa L. cv Grand Rapids) seed germination at 32 degrees C was investigated. Combinations of GA(3) (0.5 millimolar), KIN (0.05 millimolar), and CO(2) (10%) were used to induce germination. Little germination occurred in controls or upon treatment with ethylene, KIN, or CO(2). Neither KIN nor CO(2) affected the rate of ethylene production by seeds. Both germination and ethylene production were slightly promoted by GA(3). Treatments with GA(3)+CO(2), GA(3)+KIN, or GA(3)+CO(2)+KIN resulted in approximately 10-to 40-fold increases in ethylene production and 50 to 100% promotion of germination as compared to controls. Initial ethylene evolution from the treated seeds was greater than from the controls and a major surge in ethylene evolution occurred at the time of visible germination. Application of 1 millimolar 2-aminoethoxyvinyl glycine (AVG), an inhibitor of ethylene synthesis, in combination with any of above three treatments inhibited the ethylene production to below control levels. This was accompanied by a marked decline in germination percentage. Germination was also inhibited by 2,5-norbornadiene (0.25-2 milliliters per liter), a competitive inhibitor of ethylene action. Application of exogenous ethylene (1-100 microliters per liter) overcame the inhibitory effects of AVG and 2,5-norbornadiene on germination. The results demonstrate that endogenous ethylene synthesis and action are essential for the alleviation of thermoinhibition of lettuce seeds by combinations of GA(3), KIN

  7. Multiplicação in vitro DE Ficus carica L.: efeito da cinetina e do ácido giberélico In vitro multiplication of Ficus carica L.: kinetin and giberelic acid effects

    Chrystiane Borges Fráguas

    2004-02-01

    Full Text Available A cultura da figueira é afetada pelo vírus-do-mosaico e a cultura de tecidos é uma alternativa para se proceder à limpeza clonal. Neste trabalho, objetivou-se estudar o efeito da cinetina e GA3 na multiplicação in vitro da figueira. Segmentos nodais foram inoculados em meio de cultura WPM contendo as seguintes combinações de cinetina (0; 0,5; 1; 2 e 4 mg.L-1 e GA3 (0, 2, 4, 6 e 8 mg.L-1. Avaliaram-se número e comprimento dos brotos, peso da matéria fresca e seca da parte aérea, número de raízes, peso da matéria fresca e seca do sistema radicular e de calos. A utilização de 0,5 mg.L-1 de cinetina promoveu melhor taxa de multiplicação in vitro de Ficus carica. O GA3 reduziu a formação e multiplicação dos brotos e induziu ao estiolamento, à hiperidricidade, clorose e necrose apical das plântulas.The fig culture is affected by mosaic virus and the tissue culture is an alternative in the clonal cleaning. The kinetin and GA3 effects on in vitro fig multiplication was studied. Nodal segments were inoculated in WPM culture medium containing the following combination of kinetin (0, 0.5, 1, 2 and 4 mg.L-1 and GA3 (0, 2, 4, 6 and 8 mg.L-1. The number and length, fresh and dry weigh matter of aerial part, number of roots, fresh and dry weight matter of root system and fresh and dry weight matter of callus were evaluated. The use of kinetin 0.5 mg.L-1 promoted higher rates of in vitro Ficus carica multiplication. The GA3 reduced the formation and shoot multiplication, and induced etiolation, hyperhydricity, clorosis and apical necrosis at the plantlets.

  8. Embriogênese somática direta em explantes foliares de Coffea arabica L. cv. acaiá cerrado: efeito de cinetina e ácido giberélico Direct somatic embryogenesis in Coffea arabica L. cv. Acaiá Cerrado: kinetin and giberelic acid effects

    Alba Regina Pereira

    2007-04-01

    Full Text Available Objetivou-se estudar o efeito da cinetina, GA3 e ANA na indução in vitro de embriões somáticos de cafeeiro pela via direta. Segmentos foliares retirados de plântulas cultivadas in vitro foram inoculados em meio de cultura 'MS' com 50% dos sais contendo as seguintes combinações de cinetina (0; 1; 2; 4 e 8 mg L-1 e GA3 (0; 2,5; 5; 10 e 20 mg L-1. Os meios de cultura utilizados tiveram pH ajustado para 5,8 ± 1 antes de serem autoclavados. O experimento foi mantido em sala de crescimento a 25 ± 1ºC. Avaliou-se número total de embriões somáticos, o número de embriões cotiledonares, o número de embriões torpedo e a média dos comprimentos dos embriões. A ação combinada entre cinetina, GA3 e ANA estimulou a indução de embriões somáticos pela via direta. O maior comprimento de embriões foi observado quando se utilizou 8 mg L-1 de cinetina e 8,0 mg L-1 de ANA ou 17 mg L-1 de GA3 e 8,0 mg L-1 de ANA isoladamente.It was aimed to study kinetin, GA3 and ANA effects in the in vitro induction of direct somatics embryos. Leaf segments withdrawn from plantlets in vitro were inoculated in 'MS'50% containing the following combination of kinetin (0; 1; 2; 4 and 8 mg L-1 and GA3 (0; 2,5; 5; 10 and 20 mg L-1. The culture media utilized had their pH adjusted to 5,8 ± 1 before being autoclaved. The experiment was carried out growth room at 25 ± 1ºC. Total number of embryos, number of embryos cotiledonar, number of embryos of torped and length of embryo were evaluated. The combination between kinetin and GA3 promoved induction of embryos. The use of kinetin 8 mg L-1 and GA3 17 mg L-1 not associate in medium with 8,0mg L-1 of ANA, promoter higher rates in vitro.

  9. Cytotoxicity against MCF-7 breast cancer cell line and interaction ...

    N6-furfuryladenine (kinetin) is a cytokinin growth factor with several biological effects observed in human cells and fruit flies. Kinetin exists naturally in the DNA of almost all organisms tested so far, including human cells and various plants. The cytotoxicity effect of kinetin on MCF-7 breast cancer cell lines was measured by ...

  10. The role of growth regulators, embryo age and genotypes on ...

    Administrator

    2011-06-06

    Jun 6, 2011 ... 0.1 mg/l kinetin, MS + 0.1 mg/l IAA and MS + 0.1 mg/l kinetin + 0.1 mg/l IAA were used as growth regulators. ... factor for a high success in zygotic embryo culture is the ... regulators components have proved to influence the.

  11. Journal of Genetics | Indian Academy of Sciences

    Availability during embryo development of exogenously supplied cytokinin kinetin suppressed the mutant phenotype. These observations suggest that, in C. roseus,. insufficiency in endogenous kinetin may lead to monocotyledonous embryo patterning and; dicotyledonous embryo formation requires a critical amount of ...

  12. Effects of ionizing-radiation and post-radiation action of some plant growth regulators on the seed germination and seedling growth of Scotch pine

    Leszek Michalski

    2015-01-01

    Full Text Available The effects of small doses of gamma irradiation on the seed germination and seedling growth of Scotch pine and post-radiation action of water solutions of IAA, GA3 and kinetin have been investigated. Changes in the destructive action of ionizing-radiation toy gibberellic acid and its intensifying by IAA and kinetin has been found.

  13. Natural plant hormones cytokinins increase stress resistance and longevity of Caenorhabditis elegans

    Kadlecová, Alena; Jirsa, Tomáš; Novák, Ondřej; Kammenga, Jan; Strnad, Miroslav; Voller, Jiří

    2018-01-01

    Cytokinins are phytohormones that are involved in many processes in plants, including growth, differentiation and leaf senescence. However, they also have various activities in animals. For example, kinetin and trans-zeatin can reduce levels of several aging markers in human fibroblasts. Kinetin

  14. The recovery of cytokinins during extraction and purification of clubroot tissue

    Dekhuijzen, H.M.; Gevers, E.C.T.

    1975-01-01

    Losses of one naturally occurring cytokinin (zeatin) and one synthetic cytoknin (kinetin) were determined during purification of turnips (Brassica compestris) infected by Plasmodiophora brassicae (clubroot). A known amount of zeatin and 8‐14C‐kinetin was added after homogenization of plant material

  15. In vitro propagation of miracle berry (Synsepalum dulcificum Daniel ...

    SERVER

    2008-02-05

    Feb 5, 2008 ... sour lime, lemon, grape fruits and even vinegar to taste sweet (Rehm and Espig, 1991). Various studies ... 10% coconut water. T. 30% coconut water. U. 0.5 NAA + 0.2 BAP. V. 0.1 NAA + 0.3 BAP. W. 0.5 2,4–D + 0.4 Kinetin. X. 0.2 2,4–D + 0.1 Kinetin. Y. 0.02 2,4–D + 0.01 Kinetin. Z. 0 supplement (control).

  16. Germination and in vitro multiplication of Helianthemum kahiricum, a ...

    aghomotsegin

    2015-03-25

    Mar 25, 2015 ... Seeds of Helianthemum kahiricum have an excellent germination rate ... of H. kahiricum on Murashige and Skoog medium (MS) free growth regulators ... The kinetin had a positive effect on the multiplication and growth, but a.

  17. IN-VITRO PROPAGATION OF Picralima nitida (Stapf) - A threatened ...

    Dr. Gbadamosi

    plant tissue culture and micropropagation techniques play ... MATERIALS AND METHODS ... mg/L) plus 0.01 mg of α-naphthalene acetic acid (NAA); kinetin .... Summary of analysis of variance for growth variables of embryo culture sprout of P.

  18. (Catharanthus roseus) tissue culture

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... Abbreviations: NAA, Naphthalen acetic acid; Kin, Kinetin; MS,. Murashige and .... Acidic phase was isolated by sulphuric acid (5%) and ... reaction with Ceric Ammonium Sulfate (CAS) and Ultra violet (UV) detection (λ=254 ...

  19. Induced mutation to monocotyledony in periwinkle, Catharanthus ...

    Unknown

    (i) insufficiency in endogenous kinetin may lead to monocotyledonous embryo patterning and (ii) dicotyledonous em- ... Induced-mutagenesis experiments in plants have so far .... C. roseus is a small perennial herb of family Apocyna-.

  20. The effect of cytokinins on flax seed germination at low temperature

    Irena Niedźwiedź-Siegień

    2011-01-01

    Full Text Available Germination of flax seeds (Linum usitatissimum L., cv. Szafir at 5oC was enhanced by continuous white light, gibberellin A3 (GA3, kinetin and benzylaminopurine. GA3 and kinetin at physiological concentrations (10-8-10-6 M improved significantly germination in darkness. Stimulatory effect of benzylaminopurine was visible only in the light. Almost no effect of zeatin and isopentenyladenine (2iP on germination was observed. Possible causes of this differences were suggested.

  1. Role of plant growth regulators on oil yield and biodiesel production of linseed (linum usitatissimum l)

    Faizanullah, A.; Bano, A.; Nosheen, A.

    2010-01-01

    A field experiment was conducted to compare the effect of plant growth regulators (PGRs) viz. kinetin (K), chlorocholine chloride (CCC) and salicylic acid (SA) on seed yield, oil content and oil quality of Linseed (Linum usitatissimum L) cv. Chandni with a new perspective to biodiesel production. The growth regulators (10-6M) were applied as seed soaking for 10 h prior to cultivation. Kinetin significantly increased the number of capsules/plant, seed number/capsule, 1000 seed weight and total seed yield (kg/h). The growth regulators increased the seed oil content maximum being in kinetin and CCC treatments. Kinetin and CCC significantly decreased the oil acid value, free fatty acid content (% oleic acid) and increased the pH of oil. Nevertheless, SA significantly decreased the oil specific gravity and did not alter the pH. Only kinetin significantly increased the oil iodine value. The oil extracted from seeds of kinetin and CCC treated plants showed maximum conversion (% w/w) to methyl esters/biodiesel after transesterification. It can be inferred that PGRs can be utilized successfully for improving the biodiesel yield of linseed. (author)

  2. Growth of vegetative explant Moringa oleifera on different composition of auxin and cytokinin and its synthetic seed germination

    Muslihatin, Wirdhatul; Jadid, Nurul; Puspitasari, Ika D.; Safitri, Chusnul E.

    2017-06-01

    The spread of Moringa oleifera is also rare for seed germination and viability or survival are low, and the lack of vegetative propagation method. The purpose of this study are to determine the effect of auxin and cytokinin on growth vegetative explants Moringa oleifera and its synthetic seed germination. The explants grown on MS medium with sucrose content of 30% and a range of additional hormone. Addition concentration and different types of hormone made in order to know the sensitivity and response explant growth on a variety of media to get a good callus and embryosomatic. The composition of the hormone given is MS + 2.4 D 3 ppm; MS + 2,4D 2 ppm + BAP 2 ppm; MS + NAA + 0.5 ppm kinetin 1 ppm; MS + NAA 1 ppm + kinetin 1 ppm; MS + NAA 1 ppm + 0.5 ppm kinetin. The explants were incubated at a temperature of 18-20 ° C with a photoperiod 16/8. Explants and MS medium is incubated to form embryonic callus. Seeds synthetic made from embryonic callus growing on medium 1 ppm kinetin + NAA 1 ppm with encapsulation method with sodium alginate 2%. Seed synthetic germinated in some kind of medium that medium ms0 solid (M1), ms0 liquid (M2), MS0 semi-solid (M3), MS solid NAA 1ppm + Kinetin 1 ppm (M4), MS liquid NAA 1 ppm + kinetin (M5), and semi-solid MS + NAA 1 ppm kinetin 1 ppm (M6). Synthetic seed viability was observed with the parameters of the fresh weight of synthetic seed, germination percentage and seedling. Chlorophyll content was measured by spectrophotometric method with solvent asseton. Best callus generated in this study are embryonic callus that grew on media NAA 1 ppm + kinetin 1 ppm. Embryonic callus on M6 + NAA 1 ppm kinetin 1 ppm capable of germination with an average weight of callus and sprouts of 40.38 mg. Of the entire amount of a synthetic seed on M6, just 5 seed germinate, so the percentage of germination of seeds is equal to 41.67%. with an average length of sprouts 1 cm with an average total chlorophyll content of 8.66 mg / g.

  3. Micropropagation of seedless lemon (Citrus limon L. cv. Kaghzi Kalan) and assessment of genetic fidelity of micropropagated plants using RAPD markers.

    Goswami, Komal; Sharma, R; Singh, P K; Singh, Govind

    2013-01-01

    A micropropagation protocol was developed for multiplication of seedless lemon (Citrus limon L. cv. Kaghzi Kalan) using nodal explants. The maximum shoot regeneration was observed on low level of BAP (0.1 mg l(-1)) or kinetin (0.5 mg l(-1)). BA was recorded to be better than kinetin in terms of number of days taken to bud break. The maximum number of shoots per explant was observed on 0.1 mg l(-1) BA and 0.5 mg l(-1) kinetin. Shoot proliferation decreased with increasing concentration of BA alone, but in case of a combination of BA and NAA (0.1 mg l(-1) each), it increased with increasing concentration of BA up to 10.0 mg l(-1). None of the treatments including BA or kinetin alone or BA in combination with NAA produced significantly more shoots for commercial exploitation. In the case of a combination of BA + kinetin + IBA, the maximum (5.5 shoots per explants) proliferation was observed on MS medium containing 1.0 mg l(-1) BA + 0.5 mg l(-1) kinetin + 0.5 mg l(-1) IBA or 0.25 mg l(-1) BA + 1.0 mg l(-1) kinetin + 1.0 mg l(-1) IBA. Regenerated shoots showed root induction on MS basal medium or on MS medium containing 1.0 mg l(-1) IBA. It is concluded that a five-fold increase (1.0 mg l(-1) BA + 0.5 mg l(-1) kinetin + 0.5 mg l(-1) IBA) in axillary shoot proliferation, while seven-fold increase (0.25 mg/l mg l(-1) BA + 1.0 mg l(-1) kinetin + 1.0 mg l(-1) IBA) during the second cycle of multiplication could be obtained using the two plant growth regulator combinations. PCR amplification with 14 different random primers confirmed no somaclonal variant up to two cycles of shoot multiplication.

  4. The Growth and Cultivation of Meristem From Shallot (Allium ascalonicum L. cv. Katumi by In Vitro

    Lamro Purba

    2017-12-01

    Full Text Available This study aimed for knowing and obtaining the best concentration of kinetin and NAA interaction effects in influencing the shoot induction, knowing how the plant growth regulators in induction mediastill affect the shoot additionin the MS0media and also knowing the largest number of roots in rooting media for shallot by in vitro. The experiment was conducted at Laboratory of Tissue Culture Seed Technology, Faculty of Agriculture, Padjadjaran University, during January 2011 until May 2011. This experiment divided in 3 stages, namely shoot induction stage, shoot subculture to MS0 media stage and shoot subculture to rooting media stage. Experimental method used in the shoot induction stage was factorial Completely Randomized Design with three replications. The first factor was the kinetin with four levels,0, 1, 2, and 3 mg L-1. The second factor was the NAA with three levels, as 0, 0.01, and 0.1 mg L-1. Basic media used for each treatment was MS. The experiment result showed there was an interaction between kinetin and NAA on shoot induction stagewith the plantlet height, leaf number, and shoot addition. The best result for leaf number was gained from interaction with 2 mg L-1 kinetin without NAA,while the treatment of 2 mg L-1 kinetin with 0.01 mg L-1 NAA gave a better interaction for theshoot addition variable.

  5. Adventive plants from ovules and nucelli in Citrus.

    Kochba, J; Spiegel-Roy, P; Safran, H

    1972-09-01

    1- to 8-week-old ovules and nucelli from three Citrus cultivars-Shamouti and Valencia (Citrus sinensis) oranges and Marsh Seedless (C. paradisi) grapefruit-were cultured in vitro. No embryo differentiation was observed in the explants prior to culture. The Shamouti ovules had degenerated and were apparently unfertilized. Embryoids formed on Murashige and Tucker nutrient medium supplemented with 500 mg/l malt extract. Whole plants developed on the same basal medium supplemented with kinetin and indole-3-acetic acid (IAA), coconut milk or gibberellic acid (GA3). A higher kinetin/IAA ratio or the addition of coconut milk favoured stem elongation more than root formation while a lower kinetin/IAA ratio favoured root formation and inhibited stem elongation. The addition of GA3 to the basal medium stimulated rooting and stem elongation. These results can be of aid in mutation research, allowing irradiation at stages prior to embryonic development.

  6. Inhibition of ethylene production by cobaltous ion

    Lau, O.L; Yang, S.F.

    1976-01-01

    The effect of Co 2+ on ethylene production by mung bean (Phaseolus aureus Roxb.) and by apple tissues was studied. Co 2+ , depending on concentrations applied, effectively inhibited ethylene production by both tissues. It also strongly inhibited the ethylene production induced by IAA, kinetin, IAA plus kinetin, Ca 2+ , kinetin plus Ca 2+ , or Cu 2+ treatments in mung bean hypocotyl segments. While Co 2+ greatly inhibited ethylene production, it had little effect on the respiration of apple tissue, indicating that Co 2+ does not exert its inhibitory effect as a general metabolic inhibitor. Ni 2+ , which belongs to the same group as Co 2+ in the periodic table, also markedly curtailed both the basal and the induced ethylene production by apple and mung bean hypocotyl tissues. In a system in which kinetin and Ca 2+ were applied together, kinetin greatly enhanced Ca 2+ uptake, thus enhancing ethylene production. Co 2+ , however, slightly inhibited the uptake of Ca 2+ but appreciably inhibited ethylene production, either in the presence or in the absence of kinetin. Tracer experiments using apple tissue indicated that Co 2+ strongly inhibited the in vivo conversion of L-[U-- 14 C]methionine to 14 C-ethylene. These data suggested that Co 2+ inhibited ethylene production by inhibiting the conversion of methionine to ethylene, a common step which is required for ethylene formation by higher plants. Co 2+ is known to promote elongation, leaf expansion, and hook opening in excised plant parts in response to applied auxins or cytokinins.Since ethylene is known to inhibit those growth phenomena, it is suggested that Co 2+ exerts its promotive effect, at least in part, by inhibiting ethylene formation

  7. Clonal propagation of Stevia rebaudiana Bertoni by stem-tip culture.

    Tamura, Y; Nakamura, S; Fukui, H; Tabata, M

    1984-10-01

    Clonal propagation of Stevia rebaudiana has been established by culturing stem-tips with a few leaf primordia on an agar medium supplemented with a high concentration (10 mg/l) of kinetin. Anatomical examination has suggested that these multiple shoots originate from a number of adventitious buds formed on the margin of the leaf. Innumerable shoots can be obtained by repeating the cycle of multiple-shoot formation from a single stem-tip of Stevia. These shoots produce roots when transferred to a medium containing NAA (0.1 mg/l) without kinetin. The regenerated plantlets can be transplanted to soil.

  8. Hormonal effect on polyphenol accumulation in Cassia tissues cultured in vitro

    Shah, R R; Subbaiah, K V; Mehta, A R

    1976-06-01

    Effects of auxin and kinetin on growth and production of phenolic compounds in cultured Cassia fistula L. tissues were examined. Initiation of polyphenols was largely determined by the auxin concentration in the medium. Growth of the cells in relation to accumulation of polyphenols was studied at different auxin and kinetin concentrations. The accumulation of phenolic materials was essentially restricted to the most rapid phase of the growth cycle. Progressive changes in the pattern of peroxidase activity were followed and their relationship with polyphenol synthesis is examined.

  9. Hybridization of several Aerides species and in vitro germination of ...

    The wild species of Aerides namely Aerides odorata, A. odorata var.'Yellow', Aerides flabellata and Aerides quinquevulnera var. calayana are fragrant and distributed in Malaysia. Crosses among them attempted to produce primary hybrids which were used to investigate the effects of kinetin and BAP concentrations on seed ...

  10. Effect of gamma-radiation on callus initiation and oraganogenesis in the tissue culture of Nicotiana tabaccum L

    Shin, S. H.; Kim, J. G.; Song, H. S.

    2004-01-01

    It is generally agreed that ionizing radiations stimulate cell division, growth and development in various organisms including animals and plants. Differentiating tissues are the most sensitive to radiation. The present experiment was carried out to investigate the effects of ionizing radiation on callus initiation and organogenesis from the stem in the culture of Nicotiana tabaccum L. cv. When the stem segments were cultured on a Murashige and Skoog (MS) medium with 2 mg/L kinetin, with 1 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), with 2 mg/L kinetin and 1 mg/L 2,4-D, the shoots and callus were differentiated 14 days after cultivation. Callus was especially formed on the MS medium with 2,4-D and/or kinetin and the formation was promoted by 1 Gy and 5 Gy of gamma radiation. The formation of the shoot clusters on the MS medium with 2 mg/L kinetin were prominent in the 5 Gy-irradiated groups. It is concluded that that gamma radiation enhanced the callus initiation and organogenesis in the tissue culture of Nicotiana tabaccum L

  11. Abscisic Acid-Cytokinin Antagonism Modulates Resistance Against Pseudomonas syringae in Tobacco.

    Großkinsky, Dominik K; van der Graaff, Eric; Roitsch, Thomas

    2014-12-01

    Phytohormones are known as essential regulators of plant defenses, with ethylene, jasmonic acid, and salicylic acid as the central immunity backbone, while other phytohormones have been demonstrated to interact with this. Only recently, a function of the classic phytohormone cytokinin in plant immunity has been described in Arabidopsis, rice, and tobacco. Although interactions of cytokinins with salicylic acid and auxin have been indicated, the complete network of cytokinin interactions with other immunity-relevant phytohormones is not yet understood. Therefore, we studied the interaction of kinetin and abscisic acid as a negative regulator of plant immunity to modulate resistance in tobacco against Pseudomonas syringae. By analyzing infection symptoms, pathogen proliferation, and accumulation of the phytoalexin scopoletin as a key mediator of kinetin-induced resistance in tobacco, antagonistic interaction of these phytohormones in plant immunity was identified. Kinetin reduced abscisic acid levels in tobacco, while increased abscisic acid levels by exogenous application or inhibition of abscisic acid catabolism by diniconazole neutralized kinetin-induced resistance. Based on these results, we conclude that reduction of abscisic acid levels by enhanced abscisic acid catabolism strongly contributes to cytokinin-mediated resistance effects. Thus, the identified cytokinin-abscisic acid antagonism is a novel regulatory mechanism in plant immunity.

  12. Page 1 Rev. Sci. Technol., Synthese 26: 57 - 64 (2013) D. Ali ...

    Une augmentation de la teneur en proline est observee dans les plantes traitees ou non par !'hormone la kinetine. Mots cles: ... hormone in plants under stress induced a proportional increase in chlorophyll (a) and (b) with the concentration of used salt. ..... grains and vegetables, Agronomy Journal, Vol. 66,. 412-421.

  13. Effect of cytokinins on in vitro multiplication, volatiles composition and rosmarinic acid content of Thymus leucotrichus Hal. shoots.

    Bekircan, Tuba; Yaşar, Ahmet; Yıldırım, Sercan; Sökmen, Münevver; Sökmen, Atalay

    2018-03-01

    An efficient in vitro multiplication protocol was designed to Thymus leucotrichus , a subshrub and perennial herb growing naturally in the Northwest of Turkey. Of all basal media studied, Murashige and Skoog medium was found to be superior to the others, providing higher shoot formation and the maximum shoot length. Varying concentrations of cytokinins, i.e., 6-benzyladenine, thidiazuron, 2-isopentenyladenine and kinetin were supplemented in the nutrient media to observe their effects on shoot development and biomass. Rosmarinic acid content and volatile compositions of both naturally growing plants and in vitro multiplied plantlets were also evaluated. 6-benzyladenine (1.0 mg/L) and kinetin (0.5 mg/L) were found to be optimum for shoot number and shoot elongation, respectively. Thidiazuron (1.0 mg/L) was superior for biomass production. Rosmarinic acid content of in vitro multiplied plants was found to be higher than that of wild plants, reaching a maximum with 0.5 mg/L 2-isopentenyladenine, which yielded 10.15 mg/g dry weight. The highest thymol content was obtained with 1.0 mg/L kinetin (55.82%), while thidiazuron (0.1 mg/L) increased carvacrol production (12.53%). Overall, Murashige and Skoog medium supplemented with 1.0 mg/L kinetin was determined to be the most favorable medium studied.

  14. Cytokinin treatment and flower quality in Phalaenopsis orchids ...

    We previously documented an N-6-benzyladenine (BA) protocol to increase spike and flower number in Phalaenopsis orchids. To increase options for growers, we tested two additional cytokinins, kinetin (Kin) and 2-iso-pentenyl adenine (2-iP), comparing them with BA. Two key commercial cultivars were used ...

  15. Minimizing the hyperhydricity associated with in vitro growth and ...

    Hyperhydricity or glassiness is considered as a frequent problem associated with the in vitro growth and development of watermelon (Citrulus lanatus, cv. Giza 1). Explants were cultured on MS (Murashige and Skoog, 1962) medium containing 6-bensyladenine (BA), kinetin (Kin) or thidiazuron (N-phenyl N 1,2 ...

  16. Comparison of seed priming techniques with regards to germination ...

    Seeds of watermelon [Citrullus lanatus (Thunb.) Matsum and Nakai, cv. Crimson Sweet] were used to investigate the effects of different priming techniques on seed germination and early seedling growth. The seeds were soaked in solutions of 0.2% gibberellin (50 mg L–1 gibberellic acid), 0.2% cytokinin (90 mg L–1 kinetin) ...

  17. Journal of Genetics | Indian Academy of Sciences

    Home; Journals; Journal of Genetics. Sushil Kumar. Articles written in Journal of Genetics. Volume 79 Issue 3 2000 pp 97-104. Induced mutation to monocotyledony in periwinkle, Catharanthus roseus, and suppression of mutant phenotype by kinetin · Shashi Pandey Rai Sushil Kumar · More Details Abstract Fulltext PDF.

  18. Comparative analysis of hyoscine in wild-type and in vitro- grown ...

    regeneration was obtained with 2 mg/L BAP and 1 mg/L kinetin. ... Wild root, stem and leaves exhibited higher amounts (approx. ... Despite several medical benefits offered by .... The initial screening of hyoscine from all samples ... Mobile phase of 0.02 mol/L ..... compounds in commercial herbal drugs and spices from.

  19. Effect of Some Phytohormones on Growth Characteristics of ...

    IBA), gibelleric acid (GA3), and kinetin] on cell dry weight, cell number, cell size, protein and chlorophyll contents of Chlorella sorokiniana IAM-C212 were investigated under photoautotrophic conditions. Treatment with IAA (15 mg/l) and IBA (15 ...

  20. THE EFFECTS OF SEED SOAKING WITH PLANT GROWTH REGULATORS ON SEEDLING VIGOR OF WHEAT UNDER SALINITY STRESS

    Afzal Irfan

    2005-08-01

    Full Text Available Effects of seed soaking with plant growth regulators (IAA, GA3, kinetin or prostart on wheat (Triticum aestivum cv. Auqab-2000 emergence and seedling growth under normal (4 dS/cm and saline (15 dS/cm conditions were studied to determine their usefulness in increasing relative salt-tolerance. During emergence test, emergence percentage and mean emergence time (MET were significantly affected by most of priming treatments, however, root and shoot length, fresh and dry weight of seedlings were significantly increased by 25 ppm kinetin followed by 1% prostart for 2 h treatments under both normal and saline conditions. All pre-sowing seed treatments decreased the electrolyte leakage of steep water as compared to that of non-primed seeds even after 12 h of soaking. Seed soaking with 25 ppm kinetin induced maximum decrease in electrolyte leakage while an increase in electrolyte leakage was observed by 25, 50 or 100 ppm IAA treatments. It is concluded that priming has reduced the severity of the effect of salinity but the amelioration was better due to 25 ppm kinetin and 1% prostart (2 h treatments as these showed best results on seedling growth, fresh and dry weights under non-saline and saline conditions whereas seed soaking with IAA and GA3 were not effective in inducing salt tolerance under present experimental material and conditions.

  1. Radiorestoration properties of several cytokinin-like substances on the normal tissues of Jerusalem artichoke treated by Co60 gamma-rays

    Jonard, Robert; Bayonove, Jacqueline

    1976-01-01

    6 cytokinin-like substances, particularly zeatin and isopentenyladenin (IPA) contained in yeast extract and their RNA prove to be effective radiorestorative agents on Jerusalem artichoke tissues cultivated in vitro; and their restorative activity is greater than that of kinetin [fr

  2. 40 CFR 180.1157 - Cytokinins; exemption from the requirement of a tolerance.

    2010-07-01

    ... extract of seaweed meal and kinetin) in or on all food commodities when used as plant regulators on plants, seeds, or cuttings and on all food commodities after harvest in accordance with good agricultural... AGENCY (CONTINUED) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD...

  3. New cytokinin derivatives possess UVA and UVB photoprotective effect on human skin cells and prevent oxidative stress

    Hönig, Martin; Plíhalová, Lucie; Spíchal, L.; Grúz, Jiří; Kadlecová, Alena; Voller, Jiří; Rajnochová Svobodová, A.; Vostálová, J.; Ulrichová, J.; Doležal, Karel; Strnad, Miroslav

    2018-01-01

    Roč. 150, APR 25 (2018), s. 946-957 ISSN 0223-5234 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : Aromatic cytokinins * Caenorhabditis elegans * Kinetin derivatives * Oxidative stress * UVA/UVB photoprotectivity Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology Impact factor: 4.519, year: 2016

  4. Preparation, characterization and biological activity of C8-substituted cytokinins

    Zahajská, Lenka; Nisler, Jaroslav; Voller, Jiří; Gucký, Tomáš; Pospíšil, Tomáš; Spíchal, Lukáš; Strnad, Miroslav

    2017-01-01

    Roč. 135, MAR (2017), s. 115-127 ISSN 0031-9422 Institutional support: RVO:61389030 Keywords : potential purine antagonists * arabidopsis-thaliana * nucleosides * derivatives * thidiazuron * specificity * receptors * kinetin * Organic synthesis * Cytokinin bioassay * AHK3 and CRE1/AHK4 bacterial receptor assay * C8-substituted cytokinin Subject RIV: CE - Biochemistry OBOR OECD: Organic chemistry Impact factor: 3.205, year: 2016

  5. Natural plant hormones cytokinins increase stress resistance and longevity of Caenorhabditis elegans

    Kadlecová, Alena; Jirsa, Tomáš; Novák, Ondřej; Kammenga, J.; Strnad, Miroslav; Voller, J.

    2018-01-01

    Roč. 19, č. 2 (2018), s. 109-120 ISSN 1389-5729 R&D Projects: GA MŠk(CZ) LO1204; GA MŠk(CZ) LO1304 Institutional support: RVO:61389030 Keywords : Aging * Caenorhabditis elegans * Cytokinin * Kinetin * Phytohormones * Topolin * Zeatin Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 3.231, year: 2016

  6. 1896-IJBCS-Article-Abdoulaye Sereme-15 05 14

    hp

    proliferation (Chengalrayan and Gallo-. Meagher, 2001). Another suggestion would be to supply combination of cytokinins. In a study concerning Bauhinia vahlii, a leguminous plant, the combination of TDZ and kinetin recorded significant increase in shoot numbers lasting for as 4 subculture periods (Bhatt and Dhar, 2000).

  7. Microproagation of Fluted pumkin by Enhanced Axilary shoot ...

    In a bid to develop protocols for micropropagation of fluted pumkin (Telfairia occidentalis), single node cuttings of two genotypes, Telfairia occidentalis white and Telfairia occidentalis orange (ToW-1 and ToR-1 respectively) were cultured in Murashige and Skoog medium but with four different combinations of kinetin and ...

  8. Nodulation of leguminous plants as affected by root secretions and red light

    Lie, T.A.

    1964-01-01

    Nodulation of bean plants, Phaseolus vulgaris L., in water culture was poor during hot sunny weather in the greenhouse. It did not improve when indoleacetic acid, kinetin, gibberellic acid, purines and pyrimidines, yeast and soil extract were added. Nodulation was enhanced by adding used

  9. Growth Response of Explants of Irvingia Gabonensis (O'rorke, Baill ...

    Growth response of explants of Irvingia gabonensis to in vitro treatment was investigated using full, half and one quarter strength mineral components based on Murashige and Skoog medium. Plant growth regulator (kinetin-Kin) with concentration levels of 0, 1, 2, 3, 4 and 5mg/l were used for shoots initiation, while axillary ...

  10. Plant regeneration via somatic embryogenesis from root explants of ...

    A system for induction of callus and plant regeneration via somatic embryogenesis from root explants of Hevea brasiliensis Muell. Arg. clone Reyan 87-6-62 was evaluated. The influence of plant growth regulators (PGRs) including 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (6-BA) and kinetin (KT) on ...

  11. Adventitious shoot regeneration from leaf explants of the valuable ...

    The objective of this study was to develop an efficient protocol for adventitious shoot regeneration for Plectranthus barbatus Andrews using leaf explants. The explants were cultured on MS (Murashige and Skoog, 1962) medium containing various concentration of kinetin (KN), 6-benzylaminopurine (BAP) and thidiazuron ...

  12. Increasing Hermaphrodite Flowers using Plant Growth Regulators in Andromonoecious Jatropha curcas

    DASUMIATI

    2014-09-01

    Full Text Available Jatropha curcas (JC is a crop with potential for use in biodiesel. Production of biodiesel requires plant seed as raw material, so the viability of JC for use in biodiesel will dependent greatly on the plant's production of flowers. Generally, this plant is monoecious, meaning it has both male and female flowers. However, very rarely JC plants may be andromonoecious. Andromonoecious specimens of JC produce hermaphrodite and male flowers in the same plant. The number of hermaphrodite flowers per inflorescence is generally low compared to the number of male flowers. The aim of this study was to increase the proportion of hermaphrodite flowers by using plant growth regulators (PGRs in andromonoecious JC. Our experiment was conducted in Randomized Block Design (RBD with 9 treatments, namely kinetin, GA3, and IAA with concentrations of 0 ppm as a control, 50 and 100 ppm of each PGRs. The treatments were applied to stem cuttings from each plant and repeated 4 times. PGRs were applied by spraying the leaves within the buds of each plant. Applications took place weekly beginning when the plants entered flower initiating phase, until inflorescence produced. Observations were conducted during the treatment period (10 weeks. Results showed that plants treated with IAA, GA3, and kinetin at 50 and 100 ppm produced increased inflorescence per plant. The increases measured were 155.4 and 92.9% of (IAA, 120.4 and 151% (GA3, 96.6 and 51.7% (kinetin respectively. In addition, we found that application and GA3 at concentrations of 50 and 100 ppm, and kinetin at 50 ppm, increased the number of hermaphrodite flowers per inflorescence by 50%, and increased the number of hermaphrodite flowers per plant by 275.6 and 183.1% (IAA, 219.5 and 254.1% (GA3, 162.9 and 103.1% (kinetin respectively. As would be expected, the number of fruit per plant increased in those specimens treated with IAA, GA3, and kinetin at 50 and 100 ppm. The increases measured were 301.7 and 167

  13. Micropropagation and acclimatization of Stevia rebaudiana Bertoni.

    Chotikadachanarong, Kittisak; Dheeranupattana, Srisuluk

    2013-09-01

    Multiple shoot induction of Stevia rebaudiana Bertoni was studied by node explants that were cultured on solidified MS media and supplemented with 0, 1, 2, 3 and 4 mg L-1 kinetin for 4 weeks. The results showed the maximum amount of multiple shoot induction (9.31+/-4.17 shoots/explant) when cultured on MS media supplemented with 3 mg L-1 kinetin. In vitro shoots were rooted on solidified MS media supplemented with 0, 0.1, 0.5 and 2 mg L-1 Naphthaleneacetic Acid (NAA) for 4 weeks. The highest number of roots (11.18+/-1.34 roots/shoot) was detected on a concentration of 0.1 mg L-1 NAA while the high survival rate (80%) was obtained when the rooted plantlets were transferred to greenhouse conditions.

  14. The influence of auxins on the biosynthesis of isoprene derivatives in callus cultures of Vaccinium corymbosum var. bluecrop.

    Migas, Piotr; Luczkiewicz, Maria; Cisowski, Wojciech

    2006-01-01

    Callus cultures of Vaccinium corymbosum var. bluecrop were optimized for their isoprene derivatives production by supplementing Schenk-Hildebrandt (SH) medium with constant concentration of kinetin (2.32 microM) and two different amounts of selected auxins. Every auxin, except for IBA, used in 10-time higher concentration (2,4D, NAA, IAA, NOA) stimulated biosynthesis of beta-sitosterol and inhibited triterpene synthesis. Quantitative analysis of isoprene derivatives in callus biomass collected on the 25th day of the experiment proved that the analyzed callus of Vaccinium corymbosum var. bluecrop synthesized the highest amount of isoprene derivatives after subculturing on SH medium modified with 22.6 microM of 2,4D and 2.32 microM of kinetin.

  15. Abscisic acid-cytokinin antagonism modulates resistance against pseudomonas syringae in Tobacco

    Grosskinsky, Dominik Kilian; van der Graaff, Eric; Roitsch, Thomas Georg

    2014-01-01

    Phytohormones are known as essential regulators of plant defenses, with ethylene, jasmonic acid, and salicylic acid as the central immunity backbone, while other phytohormones have been demonstrated to interact with this. Only recently, a function of the classic phytohormone cytokinin in plant...... immunity has been described in Arabidopsis, rice, and tobacco. Although interactions of cytokinins with salicylic acid and auxin have been indicated, the complete network of cytokinin interactions with other immunity-relevant phytohormones is not yet understood. Therefore, we studied the interaction...... of kinetin and abscisic acid as a negative regulator of plant immunity to modulate resistance in tobacco against Pseudomonas syringae. By analyzing infection symptoms, pathogen proliferation, and accumulation of the phytoalexin scopoletin as a key mediator of kinetin-induced resistance in tobacco...

  16. Hibiscus cannabinus

    morufat

    2013-05-08

    May 8, 2013 ... myo-inositol, 0.1 mg 2, 4-dichlorophenoxy acetic acid (2, 4- D), 0.5 mg kinetin and 8 g agar per litre of ... acetic acid (NAA), 0.3 mg 6-benzyl amino-purine (BAP) and 8 g agar set at pH 5.7 and incubated at 12 h ... recorded in V400, suggesting its compatibility with equa- torial climates for optimum fibre yields.

  17. Cytokinin Activity in Water-stressed Shoots 1

    Itai, Chanan; Vaadia, Yoash

    1971-01-01

    Water stress applied to the plant shoot through enhanced evaporative demands reduced cytokinin activity in extracts of xylem exudate and leaves. This reduction resembled the changes in cytokinin activity caused by water stress applied to the root. Cytokinin activity in detached wilting leaves decreased rapidly. Recovery took place after several hours in a humid chamber. Experiments with 14C-kinetin indicated that the mechanism of the inactivation and its reversal involve a chemical transformation of the cytokinin molecule. PMID:16657585

  18. Gamma-irradiation activates biochemical systems: induction of nitrate reductase activity in plant callus.

    Pandey, K N; Sabharwal, P S

    1982-01-01

    Gamma-irradiation induced high levels of nitrate reductase activity (NADH:nitrate oxidoreductase, EC 1.6.6.1) in callus of Haworthia mirabilis Haworth. Subcultures of gamma-irradiated tissues showed autonomous growth on minimal medium. We were able to mimic the effects of gamma-irradiation by inducing nitrate reductase activity in unirradiated callus with exogenous auxin and kinetin. These results revealed that induction of nitrate reductase activity by gamma-irradiation is mediated through i...

  19. Effect of precursors on flavonoid production by Hydrocotyle ...

    Callus tissue of Hydrocotyle bonariensis was initiated from the leaf of H. bonariensis treated with 2 mg/l of 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg/l kinetin. The culture was kept at 25°C, under light (cool white fluorescent tubes, 1200 lux). To optimize the precursors to increase the production of flavonoid, different ...

  20. In vitro regeneration of some Iranian alfalfa (Medicago sativa L. genotypes via somatic embryogenesis

    Majid Shokrpour

    2014-12-01

    Full Text Available An effective in vitro regeneration system is one of the prerequisites for genetic manipulation of alfalfa (Medicago sativa L. varieties and genotypes. In this research, somatic embryogenesis of four alfalfa genotypes, 6-18 (synthetic, 4-14 (Kara Yonje- Karakozlu, 3-27 (Kara Yonje Maraghe and y-6 (Regen-SY, were investigated using leaf and petiole explants. Formation of callus and somatic embryogenesis was significantly influenced by the explant type and interaction of genotype and culture medium. Petiole explants of genotype 4-14 produced the highest yield of callus (0.406 gr fresh weight of callus. Percentage of somatic embryogenesis and the number of embryos per callus in petiole explants of genotype 4-14 was higher than those of other genotypes and explants. In genotype 6-18, the highest percentage of somatic embryogenesis was achieved on MS medium containing 5 mg/L 2,4-D and 2 mg/L kinetin. There was no significant differences between genotypes and explants in terms of embryo conversion to plantlet, and on average, 58% of somatic embryos converted to plantlet on MS medium. The petiole explants of genotype 6-18 did not exhibit somatic embryogenesis response in medium containing low ratio of 2,4-D:Kinetin (5 mg/L 2,4-D and 2 mg/L kinetin. While, these explants showed somatic embryogenesis in higher ratio of 2,4-D:Kinetin (5:1. The plantlet conversion efficiency of somatic embryos produced through this study was relatively higher and therefore, the method presented in this study could be used in alfalfa genetic manipulation and molecular studies.

  1. Effects of spermidine, proline and carbohydrate sources on somatic ...

    The in vitro main roots transverse thin cell layers of Vietnamese ginseng (Panax vietnamensis Ha et. Grushv.) were cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg.l-1 2,4- dichlorophenoxyacetic acid (2,4-D), 0.2 mg.l-1 kinetin and 0.5 mg.l-1 naphthaleneacetic acid (NAA), 30 g.l-1 sucrose and 8.5 ...

  2. The effect of growth regulators on the uptake and distribution of calcium in Golden Delicious apples

    Steenkamp, J.; De Villiers, O.T.

    1979-01-01

    45 Ca, applied to the roots of Golden Delicious apple seedlings, was readily absorbed and transported to the leaves. Application of NAAm to the leaves of seedlings significantly increased the uptake of 45 Ca, whereas the growth regulators GA 3 , kinetin, SADH, CEPA and 2,4,5-TP had no such effect. Application of NAAm to intact fruits and fruit discs also significantly increased the uptake of 45 Ca [af

  3. Development of an efficient regeneration protocol for three genotypes of brassica juncea

    Bano, R.; Khan, M.H.; Rashid, H.

    2010-01-01

    Two phytohormones, auxins (Naphthalene acetic acid and Indole acetic acid) and cytokinins (Benzyl aminopurine and Kinetin) with concentrations were used to develop an efficient regeneration protocol for 3 genotypes of Brassica juncea (UCD-635, RL-18 and NIFA RAYE). The explants were cultured on MS-medium supplemented with BAP 1.0 mgL/sup -1//NAA 0.1 mgL-1, BAP 2.0 mg L/sup -1//NAA 0.2 mg L/sup -1/, BAP 3.0 mgL/sup -1/ NAA 0.3 mg L-1 and Kinetin 1.0 mg L/sup -1/ IAA 0.1 mg L/sup -1/, Kinetin 2.0 mg L/sup -1//IAA 0.2 mg L/sup -1/, Kinetin 3.0mg L-1/IAA 0.3 mg L/sup -1/. Maximum callus production (65.55) was observed on MS medium containing with BAP 2.0 mgL-1/NAA 0.2 mg L/sup -1/. Maximum shooting (22.31) was observed BAP 3.0 mg L/sup -1//NAA 0.3 mg L/sup -1/ and KIN 3.0 mg L-1/IAA 0.3 mg L/sup -1/. Regeneration efficiency was found maximum (7.13) with BAP 3.0 mg L/sup -1//NAA 0.3 mg L/sup -1/. The three genotypes were found significantly different at p greater or equal to 0.05 in shoots production and regeneration efficiency. (author)

  4. Mikropropagasi Dendrobium “Emma Pink” (Orchidaceae pada Media Kultur In Vitro

    ASTRI NUGROHO

    2006-05-01

    Full Text Available An experiment of micropropagation of the orchid Dendrobium “Emma Pink” on the media Vacin and Went using tissue culture technique has been done. The explant was obtained from the young stem of Dendrobium “Emma Pink”. Induction stage was started by culturing the first, second, third, fourth axilar buds counted from the base of the stem and apical bud. The most potential bud to be used as a source for an explant was identified from the serial culturing of explant on the liquid media with various combinations of growth regulator kinetin and IAA. The media were placed on a shaker at 90 rpm. The second and the third axilar buds were the most potential sources of explant at 10-5 M kinetin and 5.10-7 M IAA, with the degree of success for the formation of protocorm like bodies (protocorm- like bodies were 56.25% and 43.75% respectively. The formation and multiplication of buds were obtained from protocorm-like bodies which were cultured in the solid media in the addition of 10-4 M kinetin and 5.10-6M IAA. The multiplication rate of the buds has continuously increased during the subcultures with an average multiplication rate of 3.25 buds/4 weeks, 4.59 buds/4 weeks, 6.32 buds/4 weeks and 8.80 buds/4 weeks respectively. The roots were formed when the buds were cultured in the solid media by adding 5.10-6 M kinetin and M IAA. The degree of acclimatization success of Dendrobium “Emma Pink” was 93.33% for the rooted bud and 53.33% for the unrooted bud.

  5. Propagation of Drosera rotundifolia and Drosera capensis in an in vitro Culture System

    Ileana MICLEA

    2017-11-01

    Full Text Available Drosera rotundifolia and Drosera capensis (Droseraceae are carnivorous plants grown as ornamentals and sources for homeopathic medicine. The aim of this study was to optimize nutrient and growth regulator concentrations for the in vitro propagation of these species. Half strength MS medium (1/2MS was supplemented with kinetin (0.5, 2, 5 mg/l or 6-benzyladenine (3, 5 mg/l and plantlets were transferred to 1/2MS with or without cytokinins. After 8 weeks rosette diameter, plant height, number of roots, root length were recorded and plants were cultured in full strength MS, 1/2MS or 1/2MS with 0.5 mg/l α-naphthaleneacetic acid for the same period of time. Afterwards, plant characteristics (number of roots, root length, number of shoots, number of flower stalks were assessed. For D. rotundifolia, shoot development and rosette diameter increased significantly in the medium with 0.5 mg/l kinetin and 3 mg/l 6-benzyladenine, while root development decreased. Plant growth regulator free medium was more suitable for root development than medium with α-naphthaleneacetic acid and thus supported the formation of significantly more flower stalks. For D. capensis, kinetin was detrimental for shoot development, the optimum medium for both shoot and root formation being MS without plant growth regulators.

  6. Growth, Morphology and Growth Related Hormone Level in Kappaphycus alvarezii Produced by Mass Selection in Gorontalo Waters, Indonesia

    Siti Fadilah

    2016-01-01

    Full Text Available The use of high quality seed can support the success of the seaweed cultivation. This study was conducted to evaluate the growth performance, morphology and growth related hormone level of brown strain seaweed Kappaphycus alvarezii seed produced by mass selection. Selection was performed in the Tomini Gulf, Gorontalo, based on mass selection of seaweed seed protocol with a slight modification in cut-off 10% of the highest daily growth rate. Selection was carried out for four generations. The selected 4th generation of seed was then used in cultivation performance test in the Celebes Sea, North Gorontalo, for three production cycles. The results showed that the selected K. alvarezii has higher clump weight and daily growth rate, longer thallus, more number of branches, and shorter internodes compared to the unselected control and seaweed from the farmer as external control. Furthermore, total sugar content, levels of kinetin hormone and kinetin:indole-3-acetic acid ratio were higher in selected seaweeds than that of unselected control and external control. Thus, mass selection method could be used to produce high growth of seed, and kinetin and indole-3-acetic acid play an important role in growth of K. alvarezii.

  7. In Vitro Regeneration of Foxtail Millet (Setaria italica (L. Beauv. cv. Buru Hotong

    Iriawati

    2017-09-01

    Full Text Available In vitro regeneration of foxtail millet (Setaria italica (L. Beauv. was done using basal shoot explants of 10-day old seedlings. Explants were cultured in MS basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, kinetin, 6-benzylaminopurine (BAP and 1.5 ppm NiSO4. Shoot multiplication and root induction were done in Murashige and Skoog (MS basal media. Plantlets were then acclimatized in rice husk charcoal, cocopeat, or mixed media. Results showed that MS basal medium containing 0.5 ppm kinetin, 2 ppm BAP, and 0.1 ppm 2,4-D was the optimal medium for shoot induction, in which 60% of explants developed direct shoot organogenesis. In addition, callus was optimally formed in MS medium supplemented by 1 ppm kinetin, 1 ppm BAP, and 0.5 ppm 2,4-D. Regenerated shoots spontaneously developed roots after being transferred into MS basal media without growth regulator. During acclimatization, the highest survival rate of plantlets (47% was obtained in rice husk charcoal. The developed method could be useful towards improvement of this species using in vitro tissue culture techniques.

  8. Biocompatible metal decontamination from soil using Ageratum conyzoides.

    Sharma, Virbala; Pant, Deepak

    2018-05-28

    Metal pollution in soil is a serious problem among waste landfill sites and associated environment all over the globe. Amelioration of contaminated soil by plant bioaccumulation is an important strategy to protect the soil environment. Ageratum conyzoides is a common weed species that can grow easily in any contaminating site and bioaccumulate heavy metals present in the e-waste dumping/recycling sites as a natural scavenger. Soil selected for the study was contaminated with waste cathode ray tube (CRT) and printed circuit board (PCB) powder in the concentration range of 1-10 g/kg. Soil decontamination was achieved by using weed plants with ethylene diamine tetraacetic acid (EDTA, 0.1 g/kg) and kinetin (100 μM) combination in pot experiments. Fe, Mn, Zn, and Cu accumulation was found to be highest in leaves (6.51-38.58; 0.14-73.12; 5.24-269.07; 9.38-116.59%); Pb and Cr in stem (22.83-113.41; 21.05-500%), respectively, as compared with blank. Ion chromatography was used as a tool for the measurement of essential ions present in plant under different conditions. Plants showed better growth in terms of shoot, root length, biomass weight, and chlorophyll content with the proposed combination. EDTA allows the metals available for the accumulation through possible complexation. Also, the compatibility of kinetin to manage stress in plant is found to be enhanced in the presence of EDTA due to possible π-π interaction. Metal stress condition causes the deficiency of essential ions in the plants thereby disturbing its biochemistry and results in its eventual death. EDTA-kinetin hybrid treatment was found to be compatible for metal decontamination from soil, its detoxification in plants by changing its environment and restoring the essential ions for the survival of plant.

  9. Propagation of Drosera rotundifolia and Drosera capensis in an in vitro Culture System

    Ileana MICLEA; Marius ZĂHAN

    2017-01-01

    Drosera rotundifolia and Drosera capensis (Droseraceae) are carnivorous plants grown as ornamentals and sources for homeopathic medicine. The aim of this study was to optimize nutrient and growth regulator concentrations for the in vitro propagation of these species. Half strength MS medium (1/2MS) was supplemented with kinetin (0.5, 2, 5 mg/l) or 6-benzyladenine (3, 5 mg/l) and plantlets were transferred to 1/2MS with or without cytokinins. After 8 weeks rosette diameter, plant height, numbe...

  10. The natural cytokinin 2OH3MeOBAR induces cell death by a mechanism that is different from that of the „classical“ cytokinin ribosides

    Voller, Jiří; Béres, T.; Zatloukal, M.; Kaminski, P. A.; Niemann, P.; Doležal, K.; Džubák, P.; Hajdúch, M.; Strnad, Miroslav

    2017-01-01

    Roč. 136, APR (2017), s. 156-164 ISSN 0031-9422 R&D Projects: GA MŠk(CZ) LO1204; GA MŠk(CZ) LO1304; GA MŠk(CZ) LM2015064; GA MŠk LM2015063; GA ČR GA14-19590S Institutional support: RVO:61389030 Keywords : myeloid-leukemia cells * kinetin-riboside * hl-60 cells * cancer-cells * in-vitro * apoptosis * gene * rcl * identification * lines * Phytohormone * Cytokinin * Leukemia * Cancer * Apoptosis * NCI60 panel Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 3.205, year: 2016

  11. Cytokinins and coconut water promoted abnormalities in zygotic embryo culture of oil palm

    Suhaimine Chaemalee

    2011-12-01

    Full Text Available Induction of adventitious shoot formation from mature zygotic embryo of oil palm was carried out in liquid MS mediumsupplemented with various types of cytokinins. Kinetin (KN alone at concentration of 0.5 mg/l gave the highest adventitiousshoot formation at 13.4%. However, abnormal shoots in form of inflorescence-like structure (ILS were obtained in 5mg/l KN containing medium. For coconut water a big ILSs were formed at 10.6%. Histological studied revealed that thoseinflorescences had no clear floral organs.

  12. Fatty acids, phenols content, and antioxidant activity in Ibervillea sonorae callus cultures

    Estrada-Zúñiga, M.E.; Arano-Varela, H.; Buendía-González, L.; Orozco-Villafuerte, J.

    2012-01-01

    Ibervillea sonorae callus cultures were established in order to produce fatty acids (lauric, myristic, pentadecanoic, palmitic and stearic acids) and phenolic compounds. Highest callus induction (100%) was obtained in treatments containing 2.32 or 4.65 μM Kinetin (KIN) with 2.26 or 6.80 μM 2,4-Dichlorophenoxyacetic acid (2,4-D). Highest fatty acids (FA) production (48.57 mg g-1), highest total phenol content (TPC; 57.1 mg gallic acid equivalents [GAE] g-1) and highest antioxidant activity (EC...

  13. Response of pine hypocotyl sections to growth regulators and related substances

    J. Zakrzewski

    2015-01-01

    Full Text Available Growth response of Pinus silvestris hypocotyl sections to some synthetic growth regulators and related substances was studied. Elongation of hypocotyl sections was stimulated by naphtaleneacetic acid, indole-3-acetic acid, in-dole-3-propionic acid, indole-3-butyric acid, 2,4-dichlorophenoxyacetic acid, indoleaoetic amide, indoleacetic nitrile and coumarin. Indole-3-acetic acid and naphtaleneacetic acid extended period of growth up to 16 and 24 hours, respectively. Growth was inhibited by kinetin, trans-cinnamic acid and 2,3,5-tri-iodobenzoic acid. No effect of gibberellic acid, tryptophan and biotin was observed.

  14. Responses of wild husk tomato, Physalis angulata L. to growth regulators and gamma rays on chlorophyll content and fruit yield

    Raghava, R.P.; Raghava, Nisha

    1994-01-01

    Effect of different growth regulators and gamma rays on the total chlorophyll content and fruit yield were studied in wild species of husk tomato and concluded that indole-3-acetic acid (IAA) 200 and kinetin (KIN) 10 ppm showed remarkable enhancement in both total chlorophyll content and fruit yield, while maleic hydrazide (MH) 100, 200 ppm and coumarin (COU) in all the treatments enhanced total chlorophyll content. Gamma ray treatments significantly enhanced both the parameters. Amongst all the treatments maximum fruit yield was in 20 kR of gamma rays. (author). 31 refs., 1 tab

  15. Pembentukan mother plant Bacopa australis secara In-vitro dan aklimatisasi dalam aquascape air tawar

    Media Fitri Isma Nugraha

    2017-12-01

    Full Text Available Tanaman air adalah bagian penting dari ekosistem air tawar. Salah satu spesies yang terkenal adalah Bacopa australis. Hobiis aquascape saat ini memiliki ketertarikan tinggi terhadap tanaman air dengan kualitas yang bagus dari setiap spesiesnya. Metode perbanyakan tanaman air tanpa tanah, lahan pertanian dan air perlu dilakukan untuk memenuhi keinginan tersebut. Tujuan dari penelitian ini adalah untuk mendapatkan formula media kultur jaringan dan zat pengatur tumbuh yang tepat untuk multiplikasi dalam perakitan mother plant (tanaman induk Bacopa australis, serta mendapatkan media terbaik untuk aklimatisasi. Media yang digunakan adalah media Murashige dan Skoog (MS A padat dengan perbedaan konsentrasi zat pengatur tumbuh. Perlakuan uji dalam kombinasi zat pengatur tumbuh (a 0,50 mg/L BAP + 0,50 mg/L kinetin; (b 0,50 mg/L BAP; dan (c 0,50 mg/L 2,4-D. Aklimatisasi tanaman induk dilakukan pada berbagai media antara lain 1 pasir silika + pupuk aqua soil amazonia, 2. pasir malang + pupuk aqua soil amazonia, 3 pasir silika + pupuk cair; 4 pasir malang + pupuk. Hasil yang diperoleh, yaitu formula media kultur terbaik untuk multiplikasi tunas tanaman B. australis secara in-vitro adalah media MS (A yang diperkaya dengan 0,5 mg/L BAP + 0,5 mg/L kinetin, sedangkan aklimatisasi terbaik pada media pasir malang + pupuk aqua soil amazonia. Water plant is an important part of freshwater ecosystems. One of the famous species is Bacopa australis. Today, many aquascape hobbyists have a high interest in aquatic plant species that have good aesthetic appearances. To answer this challenge, a new method in-vitro propagation of aquatic plants, planted without soil, agricultural land and water was conducted. The aim of this research was to find the best growth regulator hormon formula and aclimatisation medium, in creating the mother plant Bacopa australis. The medium used was MS (Murashige and Skoog, 1974 with different growth regulator hormon, i.e: (a 0.50 mg L-1 BAP

  16. Callus induction and plant regeneration of Ulex europaeus

    Ramírez,Ingrid; Dorta,Fernando; Cuadros-Inostroza,Álvaro; Peña-Cortés,Hugo

    2012-01-01

    A callus induction and plant regeneration protocol was developed from leaf and thorn explants for the plant Ulex europaeus. Explants were incubated on 2% sucrose half-strength Murashige and Skoog Medium (MS) with various combinations of plant growth regulators and antioxidants. The best frequency of callus and shoot formation was obtained with 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg/l x kinetin (Kin) 0.2 mg/l (DK Medium; callus induction) and zeatin (Z) 1 mg/l (DK medium; shoot induction)...

  17. Propagation and molecular characterisation of rubber tree (Hevea brasiliensis Muell. Arg) in Ghana

    Antwi-wiredu, A.

    2015-07-01

    The study was aimed at the propagation and molecular characterisation of some introduced clones of Hevea brasiliensis in Ghana. Propagation of H. brasiliensis by stem cuttings and in vitro techniques was used to study alternative procedures for mass production of rubber planting materials. Brown and green stem cuttings of Clone I and Clone II were soaked for 6 hours in 0.0-22.5g/L NAA followed by propagation in a nursery bag filled with nutrient-rich soil. Only the brown stem cuttings of H. brasiliensis survived. The % survival, length of shoots, number of roots as well as length of roots of Clone II was significantly (P<0.05) higher than Clone I. Stem cuttings treated with 15.0g/L NAA significantly (P<0.05) developed higher shoots (83.33%), number of roots (6.167), length of shoots (15.38cm) and length of roots (6.00cm) than the remaining treatments. There was significant (P<0.05) effects of NAA and Clone II in sprouting and rooting growth of the brown stem cuttings. The addition of 5.0mg/L kinetin in the MS culture medium significantly (P<0.05) enhanced higher shoot development (84.00%), number of shoots (3.60) and leaves (23.40) of the shoot-tip explants compared to other treatments. In nodal explants cultured on a medium without kinetin developed higher shoots (94.00%), height of shoot (4.80cm), number of leaves (19.20), number of shoots (6.00) and number of roots (7.00) than those with kinetin treatments. However, 7.5mg/L kinetin of the nodal culture also performed significantly after the controls. A dendrogram derived from the UPGMA distinguished the rubber clones from four areas of Ghana into two clusters. The five SSR markers showed high degree of relatedness among the rubber clones which suggested high genetic similarity (IRCA317-5, IRCA41-2, IRCA331-6, IRCA230-4, IRCA109-3, B8-23) and some degree of diversity/variation (K2-18,IRCA840-7, PB217-8, PB217-10, IRCA317-16) among the clones. Thus, clones of interest could be selected for future breeding and

  18. Micropropagation of Plantago asiatica L. through culture of shoot-tips

    Joanna Makowczyńska

    2011-01-01

    Full Text Available Shoot-tip multiplication of the medicinal species - Plantago asiatica was carried on MS medium with IAA and BAP or kinetin. Best results in micropropagation were achieved by adding 0.1 mg/dm3 IAA and 1 mg/dm3 BAP. After 6 weeks shoots were transferred to MS medium for rooting. The resulting plantlets were transferred after 8 weeks into pots and after a period of adaptation into the ground (field culture. The species Plantago asiatica was propagated in vitro by shoot-tip multiplication for the first time.

  19. Shoot regeneration of callus culture from irradiated sheed of piper nigrum L by gamma rays

    Ishak; Hutabarat, D.

    1988-01-01

    Shoot regeneration was obtained from callus that induced by irradiated seed with 25 and 50 Gy of gamma-rays and then on M.S. medium containing NAA 1 ppm and 2-ip 0.5 ppm. Irradiated seed with a dose of 25 Gy produced normal root and failed to produce shoot, but rice callus. Irradiated seed with a dose of 50 Gy pruduce callus only. Shoot differentiation occured after the callus were cultured on M.S., medium containing 2-ip 1 ppm and Kinetin 2.5 ppm. (authors). 9 refs, 3 figs

  20. In vitro propagation of lnula royleana DC

    Anna Stojakowska

    2011-01-01

    Full Text Available A micropropagation method, through axillary shoot proliferation, was elaborated for Inula royleana DC. (Asteraceae, a medicinal plant native of Himalaya. Primary explants (cotyledonary node explants and secondary explants (node explants of in vitro regenerated shoots of the plant, inoculated on MS medium supplemented with 0.1 μM NAA and 5.0 μM kinetin, regenerated 3.4 ± 1.2 and 5.1 ± 1.9 axillary shoots per explant, respectively. The regenerated shoots were easily rooting and adapting to growth in soil.

  1. Gravity-induced buds formation from protonemata apical cells in the mosses

    Kyyak, Natalia; Khorkavtsiv, Yaroslava

    The acceleration of moss protonemata development after the exit it to light from darkness is important gravidependent morphogenetic manifestation of the moss protonemata. The accelerated development of mosses shows in transformation of apical protonemata cells into the gametophores buds (Ripetskyj et al., 1999). In order to establish, that such reaction on gravitation is general property of gravisensity species, or its typical only for single moss species, experiments with the following moss species - Bryum intermedium (Ludw.) Brig., Bryum caespiticium Hedw., Bryum argenteum Hedw., Dicranodontium denudatum (Brid.) Britt. were carried out. All these species in response to influence of gravitation were capable to form rich bunches of gravitropical protonemata in darkness, that testified to their gravisensity. After the transference of Petri dishes with gravitropical protonemata from darkness on light was revealed, that in 3 of the investigated species the gametophores buds were absent. Only B. argenteum has reacted to action of gravitation by buds formation from apical cells of the gravitropical protonemata. With the purpose of strengthening of buds formation process, the experiments with action of exogenous kinetin (in concentration of 10 (-6) M) were carried out. Kinetin essentially stimulated apical buds formation of B. argenteum. The quantity of apical buds has increased almost in three times in comparison with the control. Besides, on separate stolons a few (3-4) buds from one apical cell were formed. Experimentally was established, that the gametophores buds formation in mosses is controlled by phytohormones (Bopp, 1985; Demkiv et al., 1991). In conditions of gravity influence its essentially accelerated. Probably, gravity essentially strengthened acropetal transport of phytohormones and formation of attractive center in the protonemata apical cell. Our investigations have allowed to make the conclusion, that gravi-dependent formation of the apical buds is

  2. Somatic embryogenesis and in-vitro regeneration of rice (Oryza sativa L.) cultivars under one-step and multiple-step salinity stresses

    Khattak, Mohammad S. K.; Abiri, Rambod; Valdiani, Alireza

    2017-01-01

    The present study aimed to examine the effect of one-step and multiple-step salinity stress on the somatic embryogenesis of rice cultivars within the solid and liquid (cell suspension) culture media conditions. Five rice cultivars, including Puteh Perak, Mahsuri, Basmati-370, Nona Bokra and Khari......, and significant morphological changes were observed. In contrast, the multiple-step NaCl treatment of the calli and cell suspensions led to higher growth of the cultures in the presence of NaCl compared to the controls. The solid MS media, containing 3 μM IAA and 40 μM Kinetin performed as the best media...

  3. European orchid cultivation – from seed to mature plant

    Jan Ponert

    2012-02-01

    Full Text Available We describe a method for growing orchids of the genera Dactylorhiza and Ophrys, two European members of the subfamily Orchidoideae, from seeds to mature plants using asymbiotic in vitro cultures and glasshouse pot cultures. Four media were used: two new media 1/4–2 and Mo2 and two modifications of Michl medium (Michl 1988. We also describe a highly efficient technique for seed disinfection using a syringe. We tested the effects of ethanol treatment on Anacmaptis morio (L R. M. Bateman, Pridgeon & M. W. seeds, sugar media composition on Dactylorhiza majalis (Rchb. P. F. Hunt & Summerh., Oeceoclades decaryana (H. Perrier ex Guillaumin & Manguin Garay & Taylor and Ophrys lojaconoi P. Delforge and the effect of kinetin on Dactylorhiza majalis protocorm growth. Sucrose was the best carbon source, while hexose resulted in the inhibition of protocorm development at early stages. The addition of kinetin at 10 mg/l resulted in the formation of the largest protocorms. Ethanol can have positive effect on seed germination when applied for a short time (2 min, while long-time ethanol exposure (60 min can kill the seeds.

  4. PECTIMORF and BIOBRAS-16 utilization in the potato somatic embryogenesis

    Jaime R. Hidrobo Luna

    2002-01-01

    Full Text Available With the application of PECTIMORF and BIOBRAS-16, somatic embryos were obtained in potato (Solanum tuberosum, L c.v. Desirèe, of 40 days old callus obtained from stem micropropagated plants. These were used as possible substitutes for crop regulators used in culture media for the induction of somatic embryos. The culture media was composed for 10ml.l-1 of Murashige and Skoog salt, 0.1mg.l-1 ANA, 0.1mg.l-1 kinetin, 0.5mg.l-1 thiamine, 2.5mg.l-1 cistein, 100mg.l-1 mioinositol, 20g.l-1 sucrose and 2.0g.l-1 agar. Four culture medias were tested in distinct combinations that contained different concentration of PECTIMOR and BIOBRAS-16 as substitute of auxins and cytokinins. After 90 days, the results obtained showed the possibility of substituting the auxins (0.5mg.l-1 ANA and the cytokinins (0.5mg.l-1 kinetin in the culture media, because the application of PECTIMORF at 3.2mg.l-1 and BIOBRAS-16 at 1.0mg.l-1, gave friable callus, high fresh weight (more than 1.4g and a brownish color at the end of the process, moment in which the somatic embryos of different phases, appeared at the surface of the callus. Keywords: brasinoesteroids, callus, oligopectate, somatic embryo

  5. Culture medium and growth regulators on in vitro multiplication of Psidium guajava L.

    Jorge Vilchez

    2014-01-01

    Full Text Available Guava (Psidium guajava L. cultivar `Dwarf Cuban Red 18-40 EEA' has high yields. For large-scale propagation, micropropagation is a possible solution. The aim of this study was to determine the effect of two culture media, two cytokinins and an analog brasinoesteoides (DI-31 in the in vitro multiplication of this cultivar. Two culture media (MS and WPM, three concentrations of benzylaminopurine (BAP (0.5, 1.0, 1.5 mg l-1, three of kinetin (0.5, 1.0, 1.5 mg l-1 and two DI-31 (0.01 and 0.02 mg l-1 were evaluated. The variables evaluated were: number of shoots, number of leaves, shoot length and multiplication coefficient. It was found that the type of culture medium influenced the shoot multiplication of guava. The number of shoots, shoot length and multiplication coefficient were determined by the type and concentration of cytokinin added to the culture medium. With the use of WPM culture medium with 1 mg l-1 BAP It was obtained the highest values of the variables evaluated. The use of DI-31 promoted the shoot growth without affecting the multiplication coefficient. Key words: benzylaminopurine, DI-31, kinetin, guava, micropropagation, multiplication phase

  6. The influence of fast neutron irradiation through micropropagation, calli induction and cell aggregate suspension culture of tapak dara cultivate vinca rosea linn

    Wahid, Rosmiarty A.

    2000-01-01

    Study on the influence of fast neutron irradiation toward tissue induction of apical shoot, calli of leaf and corolla as well as development of bud micropropagation using variety of MS and Gamborg (B5) which were supplemented with growth hormone 2,4-D NAA, BAP and kinetin has been carried out. Cell aggregates were obtained from modified liquid media by mixing MS macro element and Gamborg vitamin. Influence of the iow level irradiation (0,5-10 Gy) was investigated for auxiliary bud micropropagation , middle (5 - 20 Gy) for calli induction, while for call aggregates higher doses (until 30 Gy) were used. Optimum growth of bud micropropagation was stimulate at dose range between 0,5-1Gy and grown on MS supplemented whit BAP and NAAN, while for leaf and corolla calli was at 5 Gy, on MS media which was supplemented whit 1 mg/L kinetin, 10mg/L BAP and 0,5 mg/L NAA. However, neutron dose of 10 Gy decreased the induction of leaf and corolla calli. The highest radioresistance was shown by cell aggregates of leaf calli that grew prosperously up to 20Gy. Key words : fast neutron, micropropagation, tissue culture, cell culture, vinca rosea L

  7. ESTUDO DOS EFEITOS FISIOLÓGICOS IN VITRO DE DIFERENTES BASES DE MEIO DE CULTURA E REGULADORES VEGETAIS EM EUCALIPTO

    Débora de Araújo Mattos

    2017-05-01

    Full Text Available O cultivo in vitro de Eucalipto surgiu com a necessidade de clonar híbridos que possuam elevado valor econômico na indústria. Nesse contexto, objetivou-se avaliar a influência de diferentes meios de cultura (JADS e MS e citocininas (BAP, meta-Topolin e Kinetin em três concentrações (0,1; 0,2; 0,4 mg.L-1 no processo de multiplicação in vitro de dois híbridos de Eucalyptus grandis x Eucalyptus urophylla, nomeados como Clone 1 e Clone 2. Os resultados indicaram que os tratamentos em meio MS foram superiores ao meio JADS, e todos os tratamentos suplementados com Kinetin foram iguais ou semelhantes aos tratamentos sem regulador vegetal. Assim, o melhor tratamento para o Clone 1 foi em base de meio MS suplementado com 0,4 mg.L-1 de BAP, enquanto que para o Clone 2 foram os tratamentos em meio MS suplementados tanto com 0,2 mg.L-1 quanto 0,4 mg.L-1 de BAP.

  8. Radiation effect in another culture rice (Oryza Sativa L.) variety Krispo-38

    Montepeque, R.; Molina, L.G.; Lopez, J.J.; Pazos, W.; Ramirez, J.

    1993-01-01

    Seeds of the rice variety Krispo-38 were irradiated with 0, 100, 200, 300 and 400 Gray (Gy) and sown in the greenhouse. From each treatment, others containing uninucleate pollen grains were collected and cultured on a N6 agar medium with 5% sucrose concentration and supplemented with mg/l naphthalene acetic acid (NAA), 1 mg/l kinetin and 1 mg/l 2,4 dichlorophenoxyacetic acid (2,4-D). The pollen grains were induced to develop callus. The percentage of others that produced calli varied from 0.8 for the 400-Gy treatment to 3.3% for the control. The calli were transferred to N6 medium with 3% sucrose concentration, supplemented with 0.5 mg/l NAA and 1 mg/l kinetin. The percentage of calli that produced green plants varied from 1.9 for the 300-Gy treatment to 10.5 for the 200 Gy treatment. Plants developed in 2-5 weeks after callus transplant. A total of 101 green plants was obtained

  9. The combination effect of auxin and cytokinin on in vitro callus formation of Physalis angulata L. - A medicinal plant

    Mastuti, Retno; Munawarti, Aminatun; Firdiana, Elok Rifqi

    2017-11-01

    Physalis angulata L. (Ciplukan) is one member of Solanaceae that has a potential as herbal medicine. This plant grows wild in the crop fields, forest edges, etc. However, ciplukan is increasingly difficult to find recently. In vitro callus is an alternative source to produce secondary metabolite production as well as to regenerate plants through indirect organogenesis. This study aims to identify the response of hypocotyl explants on in vitro callus formation induced by a combination of auxin and cytokinins. Two types of cytokinins, Kinetin and BAP (0.5 ppm) were combined with three types of auxin, i.e. 2.4-D, IBA and IAA, at three concentrations 0.5, 1.0 and 1.5 ppm. In all combinations of cytokinin and auxin, 50-100% of hypocotyl explants derived from in vitro seedling were able to produce callus either in a compact or watery friable texture. In MS medium supplemented with 2.4-D, callus FW (fresh weight) began to decline in the fourth week after culture. Callus FW that increased until 5 weeks of culture was obtained in medium IAA 0.5 + Kin 0.5, IBA 1.0 + Kin 0.5 and IBA 1 + BA 0.5. Almost all calli induced on a medium + Kinetin also produced roots. While medium + BAP was able to induce shoots regeneration.

  10. In vitro propagation of two Iranian commercial pomegranates (Punica granatum L.) cvs. 'Malas Saveh' and 'Yusef Khani'.

    Valizadehkaji, Babak; Ershadi, Ahmad; Tohidfar, Masoud

    2013-10-01

    An efficient in vitro propagation is described for Punica granatum L. using shoot tip and nodal explants. The influence of two basal medium, WPM and MS, and different plant growth regulators was investigated on micropropagation of the Iranian pomegranate cultivars, 'Malas Saveh' and 'Yousef Khani'. For proliferation stage, media supplemented with different concentrations (2.3, 4.7, 9.2 and 18.4 μM) of kinetin along with 0.54 μM NAA was used. WPM proved to be more efficient medium compared to MS. The best concentrations of kinetin were 4.7 μM for 'Malas Saveh' and 9.2 μM for 'Yousef Khani', resulting in the highest number of shoots per explants, shoot length and leaf number. For both cultivars, half-strength WPM medium supplemented with 5.4 μM NAA was most effective for rooting of shoots. Rooted plantlets were successfully acclimatized and transferred into soil. The micropropagated plants were morphologically uniform and exhibited similar growth characteristics and vegetative morphology to the mother plants.

  11. The exploit of cereal embryo structure for productive reasons by in vitro techniques

    Savaskan, C.

    2017-07-01

    There are two main sides of our works exploiting embryo structure in durum wheat and some other cereals. First is haploid (or doubled haploid) embryo production using anther or microspore culture or intergeneric crosses, to ameliorate desirable characters genetically homozygote. Secondly, to develope convenient embryo culture technique in order to be stored and cultivated longer time of genotypes without being alien pollination etc. in field conditions. For that reason, two different auxin and also their combination with kinetin were used for mature embryos of wheat genotypes (hexaploid and tetraploid), to understand efficient dose for calli production and plant regeneration in plant tissue culture. Modified MS media were used adding a single dose of arabinogalactan protein (AGP) and without adding for regeneration. In further step of this study, most efficient auxin+kinetin combination which is determined previous research, it was used in the same modified MS medium to produce calli production and plant regeneration in three different genotypes (hexaploid and tetraploid wheat and diploid barley). Data were calculated in five different developmental stages of treatments. All statistical analysis of data were performed and means were compared with Duncan's test. Genetics and morphological effects of AGP on genotypes were discussed with the results of variance analysis. Simple correlation coefficient (r) was calculated base on the main values of replications.

  12. Wheat seed enhancement by vitamin and hormonal priming

    Khan, M.B.; Gurchani, M.A.; Hussain, M.

    2011-01-01

    Seed priming has proven beneficial in many important agricultural crops. The present study was conducted to explore the role (if any) of hormonal and vitamin seed priming to improve the germination, seedling emergence, early seedling establishment, electrolyte leakage and nutrients uptake in wheat seedlings. The wheat seeds were soaked for 48 hours in aerated solution of salicylic acid, ascorbic acid, kinetin and GA3 with 20 ppm concentration of each solution, whereas untreated seeds were taken as control. Seed priming with ascorbic acid resulted in maximum final germination and emergence percentage (FGP and FEP), radical and plumule length, root and shoot length, number of secondary roots, root shoot ratio, root dry weight, shoot dry weight and seedling dry weight compared to control (untreated seeds). Minimum mean germination and emergence time (MGT and MET) was recorded in seeds primed with kinetin and GA3. While in case of biochemical attributes, seedling potassium contents were decreased by hormonal seed priming while total soluble sugars were increased by salicylic acid and ascorbic acid seed priming. Hormonal seed priming had non-significant effect on phosphorus seedling contents. Untreated (Control) seeds showed maximum electrical conductivity at 0.5, 1, 2, 6, 12 and 24 h after inbibition than primed seeds against the minimum electrical conductivity that was recorded in seeds primed with salicylic acid and ascorbic acid. In conclusion, the wheat seeds primed with 20 ppm solution of ascorbic acid may be used for wheat seed invigoration. (author)

  13. A 9-vinyladenine-based molecularly imprinted polymeric membrane for the efficient recognition of plant hormone 1H-indole-3-acetic acid

    Chen Changbao; Chen Yanjun; Zhou Jie; Wu Chunhui

    2006-01-01

    9-Vinyladenine was synthesized as a novel functional monomer for molecular imprinting techniques and its structure was established with elemental analysis and 1 H NMR spectroscopy. The binding mechanism between this functional monomer 9-vinyladenine and the plant hormone 1 H-indole-3-acetic acid in acetonitrile was studied with UV-vis spectrophotometry. Based on this study, using 1 H-indole-3-acetic acid as a template molecule, a specific 9-vinyladenine-based molecularly imprinted polymeric membrane was prepared. Then, the resultant polymeric membrane morphologies were visualized with scanning electron microscopy, and the membrane permselectivity for 1 H-indole-3-acetic acid, 1 H-indole-3-butyric acid and kinetin was tested with separate experiments and competitive diffusion experiments. These results showed that the imprinted polymeric membrane prepared with 9-vinyladenine exhibited higher transport selectivity for the template molecule 1 H-indole-3-acetic acid than 1 H-indole-3-butyric acid or kinetin. The membrane prepared with 9-vinyladenine also took on higher permselectivity for 1 H-indole-3-acetic acid in comparison with the imprinted membrane made with methacrylic acid. It is predicted that the 9-vinyladenine-based molecularly imprinted membrane may be applicable to the assay of 1 H-indole-3-acetic acid or for the preparation of a molecularly imprinted polymer sensor for the analysis of 1 H-indole-3-acetic acid in plant samples

  14. Robust regeneration protocol for the Agrobacterium tumefaciens mediated transformation of Solanum tuberosum

    Abbasi, A.; Bilal, M.; Hussain, J.; Shah, M. M.; Hassan, A.

    2016-01-01

    Plant genetic transformation requires robust regeneration system. Plant growth regulators (PGRs) such as cytokinins (CKs) play a pivotal role in organogenesis; however, CKs are the most expensive PGRs. In the current study, an efficient yet economical protocol for regeneration of potato plant was developed. Stem inter-nodal and leaf explants were cultured on different regeneration media supplemented with varying concentration of different CKs such as kinetin and zeatin. Murashige- Skoog media added with zeatin (1, 1.5 mg/L) was designated as RZ1, RZ1.5, respectively or kinetin (1.5, 2 mg/L) was designated as RK1.5 and RK2, respectively, however, concentrations of other hormones such as NAA (1-Naphthaleneacetic acid) and GA3 (Gibberellic acid A3) were kept same. RZ1 and RZ1.5 gave significantly better Results as compared to RK-type media in all aspects studied such as callus initiation, days to first shoot emergence, number of shoots per explants. RZ1 medium was then selected as regeneration media for Agrobacterium-mediated transformation of potato plants with cyanobacterial phosphoenol pyruvate carboxylase gene, which provided multiple putative transformants on selection media. The transformants were further confirmed through PCR. The current protocol is found to be cost effective and efficient for the regeneration of Solanum tuberosum and can be successfully implied for the Agrobacterium-mediated transformation. (author)

  15. EFFECT OF AUXIN AND CYTOKININ ON VINCRISTINE PRODUCTION BY CALLUS CULTURES OF CATHARANTHUS ROSEUS L. (APOCYNACEAE

    Chinnamadasamy Kalidass

    2009-11-01

    Full Text Available Callus cultures of Catharanthus roseus L. were established to verify whether they produce vincristine as the intact plant. Different growth regulator combinations were applied to Murashige and Skoog (MS medium to influence the level of production of vincristine. The effects of various combinations (0.5 µM to 3.0 µM of auxin and cytokinin on the growth and accumulation of vincristine were investigated. MS medium supplemented with 2,4-Dichlorophenoxy acetic acid (2,4-D 1.0 µM and 6-furfurylaminopurine (Kinetin 1.0 µM was used to support the growth of callus cultures and the maximum amount of dry biomass (598.04 mg was produced after seven weeks of culture. High performance liquid chromatographic (HPLC analysis of methanol extracts from callus cultures of C. roseus revealed that the cultures produced vincristine. The concentrations of the growth regulators alpha-naphthalene acetic acid (NAA and kinetin played a critical role in the production of vincristine.

  16. Direct somatic embryogenesis in Swietenia macrophylla King

    Raúl Collado

    2006-04-01

    Full Text Available Swietenia macrophylla King is difficult to be propagated by tissue culture and there is not an efficient system via organogenesis, due to problems of microbial contamination, phenolic oxidation and death of tissue in the phase of in vitro establishment of explants. In order to establish a protocol for obtaining somatic embryos, zygotic embryos were used as initial plant material. Three combinations of 2,4-D with kinetin were studied, to obtain the formation of somatic embryos. After six weeks of culture, the number of explants with high and low somatic embryogenesis frequency were determined. So that the somatic embryos in globular stage reach the final stages of torpedo and cotyledonal, these were placed in three treatments with 6-BAP (0.2, 0.4 y 0.6 mg.l-1. The number of somatic embryos that reached the torpedo and cotyledonal stages were evaluated after 30 days of culture. Results demonstrated that direct somatic embryogenesis from immature zygotic embryos is obtained in the culture medium composed by MS salts with 4.0 mg.l-1 of 2,4-D and 1.0 mg.l-1 of kinetin. Higher percentage of somatic embryos in cotiledonal stage (91.7 %, was obtained with 0.4 mg.l-1 of 6-BAP. Key word: forestry, growth regulator, mahogany, somatic embryo, tissue culture

  17. KAJIAN INDUKSI KALUS RUMPUT LAUT Kappaphycus alvarezii UNTUK PRODUKSI EMBRIO SOMATIK

    Muh. Alias L. Rajamuddin

    2016-11-01

    Full Text Available Untuk mendukung program transgenesis pada rumput laut, embrio somatik dapat digunakan sebagai material untuk transfer gen baik secara individu sel ataupun kluster sel embriogenik, sehingga mempercepat keberhasilan dengan peluang transformasi yang lebih tinggi. Penelitian ini bertujuan untuk mengkaji induksi kalus rumput laut K. alvarezii untuk produksi sel embrio somatik (e.s. dengan beberapa rasio zat pengatur tumbuh (ZPT dan konsentrasi agar media induksi, sampai sel menjadi filamen. Penelitian terdiri atas dua tahap: Tahap (1 induksi kalus, dengan rasio ZPT asam indol asetat (IAA:kinetin = 0,5:0,0 mg/L; 1,0:1,0 mg/L; dan 2,0:0,2 mg/L dengan konsentrasi agar media induksi = 0,6%; 0,8%; 1,0%; dan 1,5%. Tahap (2 regenerasi massa sel e.s., dengan rasio IAA:kinetin = 0,1:1,0 mg/L; 0,0:0,1 mg/L dan tanpa ZPT dengan konsentrasi agar media = 0,4%; 0,6%; dan 0,8%. Untuk perkembangan sel-sel e.s. lebih lanjut dipelihara pada kultur cair. Hasil penelitian menunjukkan pada tahap induksi kalus, rasio IAA: kinetin = 1:1 mg/L dengan konsentrasi agar media 0,8% dan 1,0% menghasilkan persentase induksi kalus tertinggi (90%. Pada tahap regenerasi massa sel e.s., ZPT tidak berpengaruh terhadap perkembangan massa sel e.s., di mana tanpa ZPT dengan konsentrasi agar 0,6% memperlihatkan perkembangan tertinggi (rata-rata diameter massa sel 5 mm. Pada media cair, perkembangan sel e.s. dari single cell ukuran 3-4 mm menjadi filamen-filamen ukuran rata-rata 0,5 mm dapat dicapai dalam satu bulan kultur. Keberhasilan produksi sel e.s. K. alvarezii, selain sebagai material untuk transfer gen juga dapat dijadikan acuan dalam produksi benih rumput laut kultur jaringan. To support the program of seaweed transgenesis, somatic embryo can be used as a materials for gene transfer purpose either by individual or cluster of cells in accelerating the higher rate of transformation. This research aims to study the callus induction of seaweed K. alvarezii for production of somatic

  18. In vitro and genetic diversity studies of twelve accessions of aerial yams (D. bulbifera and D. alata) in Ghana

    Katsekpor, C.

    2014-07-01

    Aerial yams (D. bulbifera and D. alata) are usually among the dioscoreaceae, in that they are cultivated not only for their tubers but also for the bulbils that develop at the leaf axils of the vine. Twelve accessions collected from five geographic regions were evaluated for morphological characteristics. The results indicate that the 12 accessions exhibited similarity with respect to 11 traits but were variable with respect to 5. Bulbils from the twelve accessions of aerial yams collected from the experimental field, were analysed for proximate composition including moisture, crude protein, crude ash, crude fat, crude fibre and carbohydrate content using the Association of Official Analytical Chemist (AOAC) method. Elemental composition of the bulbils was also determined through instrumental neutron activation analysis (INAA). Accession S recorded the highest crude protein (6.77 %), crude fibre (3.38 %) and carbohydrate (25.39 %) contents. Bulbils from D, G and T recorded the highest crude fat (0.56 %), moisture (72.97 %) and crude ash (7.96 %) content respectively. Accession V recorded the highest amount of magnesium (143.43 mg/100g), potassium (2644.4 mg/100g), chlorine (3272.2 mg/100g) and copper (1.116 mg/100g). Bulbils from accession F also recorded the highest amount of aluminum (5.26 mg/100g) and manganese (0.43 mg/100g), while accession B, C, E and R also recorded highest percentages of vanadium (0.037 mg/100g), sodium (26.63 mg/100g), calcium (333.5 mg/100g) and zinc (4.53 mg/100g) respectively. Nodal explants of 12 accessions of the aerial yam were cultured in vitro on Murashige and Skoog (MS) medium supplemented with varying concentrations of kinetin with or without activated charcoal, as assessed for ability to regenerate plantlets with distinct shoots, leaves and roots. The highest regeneration of plantlets was achieved at kinetin concentration of 2.0 mg/l with activated charcoal. Kinetin concentration beyond 2.0 mg/l proved to be phytotoxic and

  19. Gravi-photomorphogenesis of the moss Pottia intermedia protonemata

    Demkiv, O. T.; Kyjak, N. Y.; Khorkavtsiv, Y. D.; Kit, N. A.

    The protonemata development proceeds in the process of gradual differentiation of growing apical cells and intercalar cells the shortened lateral branches of the latters being transformed into three-dimensional gametophore buds (Demkiv et al., 1991). Normal course of plant development needs favourable external conditions. Sometimes, however, external environment agents can accelerate the development of organism. So, apical protonema cells of darkgrown gravitropic P. intermedia differentiate gametophore-buds in light of low intensity (Ripetskyj, 1999). We investigate the influence of gravistimulation on bud formation in haploid and diploid P. intermedia protonema. Diploid protonema was found to react on light weaker than haploid one. Under the influence of light the darkgrown apical cells and lateral branches of haploid protonema were directly transformed into buds, while in diploid protonema at first the formation of bundles of rhizoid type filaments takes place on the tips of caulonema and buds appeared in center of such bundles. The participation of gravity in gametophore bud formation was assessed by clinorotating protonema in darkness. Being illuminated such protonema also developed buds quickly the latters being formed along all stolon. It can be suggested that at 1g the growth zone of apical cells actively attract inductors of bud formation. During clinorotation the inductors probably are transferred much more slower than under stationary state and that is why the buds arised not only at the tips of stolons but along all their length. It is known that gametophore bud formation can be stimulated by exogenous phytohormones. As M. Bopp (1980) has shown, that kinetin selectively promotes bud formation on moss protonema. Our observations have shown 0,5 -- 50 μ M of kinetin stimulate the bud formation on diploid aposporic protonema much more effectively that on haploid one. It can be concluded that the amount of endogenous cytokinins in haploid protonemal cells is

  20. OPTIMIZATION OF HORMONE COMPOSITION OF NUTRIENT MEDIUM FOR IN VITRO EFFICIENT REGENERATION OF BREAD WHEAT

    E. D. Nikitina

    2016-08-01

    Full Text Available Optimal values of phytohormones in the differential nutrient medium providing the efficient realization of morphogenetic potencies of four spring bread wheat varieties (Skala, Spectr, Zarnitsa and Zhnitsa from immature embryo cultures have been determined. For callus induction explants 1.5 – 1.7 mmin size were used, which were subsequently passed to the medium by Linsmaier&Skoog possessing 0.8 % of agar, 3 % of sucrose and 2.0 mg l-1 dichlorophenoxyacetic acid (2,4-D. Cell cultures were incubated in darkness at the temperature 26±1 °С. 30 – 35 days after in accordance with the scheme of complete factorial experiment of 32 type calli were passed to differential medium supplemented with 2,4-D at levels 0.5; 2.5; 4.0 mg l-1 and with kinetin (6-furfurylaminopurine at levels 0.5; 2.25 and 4.0 mg l-1. Number of replications for each of 9 variants was four. As a result, 20 mathematic models (4 varieties × 5 stages of regeneration designed as polynomial quadric equation were obtained. On the ground of the analysis of models it was established that optimal values for factors are not equal both for cultures of genotypes analyzed and for different regeneration stages. For callus tissues of Skala and Spectr an optimal value of kinetin for all regeneration stages was 0.5 mg l-1 except for the frequency of morphogenesis. Optimal values of 2,4-D for the same varieties were within 2.3 – 3.2 mg l-1. For cell cultures of Zarnitsa and Zhnitsa recommended concentration intervals made up 1.3 – 2.2 mg l-1 on kinetin except for the frequency of rhizogenesis, and 1.9 – 2.7 on 2,4-D. The level of exogenous phytohormones necessary for stem differentiation was lower than the one for root formation. The dependence of morphogenesis results on the hormonal status of the explant has been discussed.

  1. The study of genotype, cold pretreatment, low-dose gamma irradiation and 2,4-D concentration effect on wheat doubled haploid production

    Naserian, B.; Vedadi, C.; Karbalaii, S.

    2005-01-01

    Full text: In this study, response of a cultivar (Atrak) and two lines (F3 2005 and F3 2104) of wheat , and effect of cold pretreatment, low dose gamma irradiation and 2,4-D (2,4 mg l -1 ) were investigated to anther culture response. Donar plants were grown under field conditions in early spring. Anthers were plated on modified CHB medium containing 2, 4-D (2, 4 mg l -1 ), 0.5 mg l -1 Kinetin and 90 g l -1 Sucrose. Percent of Callus formation in 100 anther and percent of plantlet in 100 calli were measured. Results indicated that genotype, cold pretreatment and 2,4-D concentrations had significant effect on anther culture response. F3 2005 showed highest callus induction and plantlet production and F3 2104 had lowest response. This experiment indicated that androgenic traits are controlled by genotype and environmental factors. Further more these traits are controlled independently. (author)

  2. Micropropagation and cytogenetic assessment of Zingiber species of Northeast India.

    Das, Archana; Kesari, Vigya; Rangan, Latha

    2013-12-01

    An improved micropropagation protocol was developed for Zingiber moran and Z. zerumbet, two wild species of the genus Zingiber, found in Northeast India. The effects of growth regulators, sugar concentrations, and nutrients were tested on the rate of shoot initiation and multiplication. An increase in proliferation and multiplication occurred in modified Murashige and Skoog (MS) medium supplemented with benzyladenine and kinetin. About 2 % sucrose and 0.7 % agar were found to be the optimum for shoot multiplication and regeneration. Naphthalene acetic acid at 0.5 mg/L produced the best rooting response for both the species. Regenerated plantlets were acclimatized successfully and cytogenetic stability was confirmed by RAPD profiling and ploidy checks.

  3. Micropropagation of Dalbergia sissoo Roxb. through tissue culture technique.

    Sahu, Jyoti; Khan, Shagufta; Sahu, Ram Kumar; Roy, Amit

    2014-04-01

    Multiple shoots of Dalbergia sissoo Roxb. (Sissoo) were incited from seeds through indirect somatic embryogenesis method. Seeds were inoculated in Murashige and Skoog's medium without any growth hormone. Than cotyledonary leaves were struck and used for callus induction on MS medium amplified with 2, 4-dichlorophenoxyacetic acid (0.5 to 4 mg mL(-1)). After 3 to 4 weeks the embryogenic callus clumps was transferred to medium supplemented with cytokinin (BAP 1 to 5 mg L(-1), kinetin 1-5.0 mg L(-1)) for embryo maturation and germination. The high-frequency shoot proliferation (82%) and maximum number of shoots per explants were recorded in MS medium containing NAA (0.5)+BAP (0.5). The findings of recent investigations have shown that, it is possible to induce indirect somatic embryogenesis in Dalbergia sissoo and plant regeneration from callus cultures derived from cotyledonary leaves as explants.

  4. In vitro technique for selection of radiation induced mutants of garlic

    Zhen, H.R.

    2001-01-01

    In vitro culture and radiation techniques were used for obtaining mutants tolerant to high temperature stress in garlic. Callus cultures and globular bodies were initiated from young leaves on MS medium containing 500 mg/l casein hydrolysate, 1000 mg/l yeast extract and 2mg/l 2,4-D. Globular bodies, resembling compact nodular calli, were irradiated with 5 to 10 Gy gamma rays. Plants and roots were obtained from globular bodies cultured on MS medium containing 2mg/l kinetin and 0.5 mg/l IAA. The plants produced bulblets (small bulbs) when cultured on MS medium containing IBA 2 mg/l and maintained at 32 deg. C. (author)

  5. Micropropagation of PLUCHEA LANCEOLATA (Oliver

    M. Kher Mafatlal

    2014-09-01

    Full Text Available Pluchea lanceolata is an important medicinal plant of Asteraceae family known for its anti-arthritic and anti-inflammatory activity. A protocol was established for micropropagation of P. lanceolata using nodal explants. Nodal explants were inoculated onto Murashige and Skoog (1962 - MS medium supple–mented with 6-benzylaminopurine (BAP, kinetin (Kin, thidiazuron (TDZ and 2iP (2-isopentenyladenine at various concentrations (0.0, 0.5, 1.0, 1.5 and 2.0 mg·dm-3. The highest multiplication rate was obtained for nodal explants cultured on MS medium, supplemented with 0.5 mg·dm-3 thidiazuron (TDZ. In vitro raised shoots were successfully rooted on ½ mineral salt concentration of MS medium supplemented with 1.0 mg dm-3 IBA.

  6. Effect of plant growth regulators and genotype on the micropropagation of adult trees of Arbutus unedo L. (strawberry tree).

    Gomes, Filomena; Simões, Mafalda; Lopes, Maria L; Canhoto, Jorge M

    2010-12-31

    Arbutus unedo grows spontaneously around the Mediterranean basin. The species is tolerant to drought and has a strong regeneration capacity following fires making it interesting for Mediterranean forestation programs. Considering the sparse information about the potential of this fruit tree to be propagated in vitro, a project to clone selected trees based on their fruit production was initiated a few years ago. The role of several factors on A. unedo propagation was evaluated. The results showed that 8.9 μm kinetin gave the best results although not significantly different from those obtained with benzyladenine or zeatin. The inclusion of thidiazuron or 1-naphthaleneacetic acid promoted callus growth and had deleterious effects on the multiplication rate. The genotype of the donor plants is also a factor interfering with the multiplication. The results also indicated that the conditions used for multiplication influenced the behavior of shoots during the rooting phase. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Distribution of Trans-Anethole and Estragole in Fennel (Foeniculum vulgare Mill of Callus Induced from Different Seedling Parts and Fruits

    Abd El-Moneim Mohamed Radwan AFIFY

    2011-03-01

    Full Text Available In the present study, seeds from local cultivar of fennel were germinated on Murashige and Skoog medium (MS without plant growth regulators. Different types of explants from the growing seedling such as cotyledonal leaves, hypocotyls, epicotyls and roots were cultured on MS medium, contained different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D either alone or with kinetin. Differential responses in the essential oil constituents were observed in the induction and development of callus. The major components of essential oils includes estragole, trans-anethole, limonene and fenchone were studied under different conditions to find out the best methods which could be used to reduce the amount of estragole (not favorite for human consumption and increase the amount of trans-anethole.

  8. Micro propagation of Stevia rebaudiana Bertoni through temporary immersion bioreactor system

    Norazlina Noordin; Rusli Ibrahim; Nur Hidayah Sajahan; Siti Maryam Mohd Nahar; Siti Hajar Mohd Nahar

    2012-01-01

    Stevia rebaudiana Bertoni is a perennial herb that belongs to the family of Asteraceae. It is a natural sweetener plant known as sweet leaf, which is estimated to be 300 times sweeter than cane sugar. In this study, micro propagation of this natural herb via temporary immersion bioreactor system was successfully conducted. Shoot tips and nodal segment were used as explants to induce multiply shoots. It was found that shoot tips on MS medium supplemented with 1 mg/l Kinetin showed the highest shoot multiplication after 3 weeks of culture. Shoot elongation and rooting was successfully optimized in MS basal medium 2 weeks later. Mass propagation of stevia shoots were carried out in temporary immersion bioreactor and this system showed promising potential as an alternative approach for rapid and continuous production of in vitro stevia plantlets. (author)

  9. Chemical regulation of sex expression in certain olive cultivars

    E. S. Hegazi

    2013-12-01

    Full Text Available The modifying effect of growth regulators on bud burst, flower bud formation and sex expression in olives varies greatly according to cultivar, concentration and time of application. Cycocel 200, 500 mg/l, ethephon 200 mg/l and SADH 2000 mg/l stimulated bud burst and flower bud formation in the 'Blanquetta' cv. A noticeable increase in the percentage of perfect flowers was obtained by 100 mg/l of kinetin, and 200 mg/l of Cycocel in the cv. 'Picual', Ethephon 200/1, Cycocel 200 mg/l and SADH 2000 mg/l considerably increased the percentage of perfect flowers in the cvs 'Blanquetta', 'Serrana' and 'Picual'. Treatments at green cluster stage were not effective.

  10. Complex, non-monotonic dose-response curves with multiple maxima: Do we (ever) sample densely enough?

    Cvrčková, Fatima; Luštinec, Jiří; Žárský, Viktor

    2015-01-01

    We usually expect the dose-response curves of biological responses to quantifiable stimuli to be simple, either monotonic or exhibiting a single maximum or minimum. Deviations are often viewed as experimental noise. However, detailed measurements in plant primary tissue cultures (stem pith explants of kale and tobacco) exposed to varying doses of sucrose, cytokinins (BA or kinetin) or auxins (IAA or NAA) revealed that growth and several biochemical parameters exhibit multiple reproducible, statistically significant maxima over a wide range of exogenous substance concentrations. This results in complex, non-monotonic dose-response curves, reminiscent of previous reports of analogous observations in both metazoan and plant systems responding to diverse pharmacological treatments. These findings suggest the existence of a hitherto neglected class of biological phenomena resulting in dose-response curves exhibiting periodic patterns of maxima and minima, whose causes remain so far uncharacterized, partly due to insufficient sampling frequency used in many studies.

  11. The effect of some growth regulators on enzyme systems in irradiated barley grain using disinfestation doses

    Bachman, S.

    1973-01-01

    Disinfestation doses of 20 to 100 krad may cause changes in the biological systems of barley grain and, therefore, may influence undesirably the technological quality of malted grain. The effect of some growth regulators on irradiated grain has been investigated. The experiments have been carried out on brewery barley var. Visa Breuns. Following growth-regulators were used: gibberellic acid (Polish preparation ''Gibrescol''), kinetin (6-furfurylo-aminopurin), CCC (2-chloroethyl trimethyl ammonium chloride), and betaine hydrochloride. By treating the irradiated barley with solutions of growth regulators it was possible to diminish the loss of enzyme activity. A ''regenerating'' effect of growth substances, mainly gibberellic acid and betain hydrochloride in 10 -4 M solutions, was observed. Amylolytic activity decreased immediately after irradiation but in samples treated with growth regulators it was higher than in those without regulators. The results may have a practical importance since gibberellic acid has just been introduced into the brewery industry. (F.J.)

  12. Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants.

    Chen, M H; Wang, P J; Maeda, E

    1987-10-01

    The regeneration potential of shoot tip, stem, leaf, cotyledon and root explants of two papaya cultivars (Carica papaya cv. 'Solo' and cv. 'Sunrise') were studed. Callus induction of these two cultivars of papaya showed that the shoot tips and stems are most suitable for forming callus, while leaves, cotyledons and roots are comparatively difficult to induce callus. Callus induction also varied with the varities. Somatic embryogenesis was obtained from 3-month-old root cultures. A medium containing half strength of MS inorganic salts, 160 mg/l adenine sulfate, 1.0 mg/1 NAA, 0.5 mg/1 kinetin and 1.0 mg/1 GA3 was optimal for embryogenesis. The callus maintained high regenerative capacity after two years of culture on this medium. Plants derived from somatic embryos were obtained under green-house conditions.

  13. Effect of ethanol and plant growth regulators on termination of potato microtuber dormancy

    Wróbel Sławomir

    2015-12-01

    Full Text Available The duration of dormancy varies significantly among cultivars, but even short dormancy can limit usage of potato microtubers for seed production. The aim of the research was to test efficacy of dormancy breaking by treatment with an aqueous solution of ethyl alcohol, saccharose, gibberellic acid and kinetin (ethanol treatment in comparison to treatment with aqueous solution of thiourea, daminozide and gibberellic acid (standard treatment. Prolonging the period of microtuber production at the in vitro stage significantly favored the short-ening of the dormancy and facilitated its breaking. While the standard treatment had the strongest effect, the ethanol treatment was slightly less efficient. The statistically significant differences were only observed during the first 13 days after the microtuber treatment. After that time, efficacy of ethanol and standard treatments was similar to control treatment with water. The investigated treatments had no effect on the natural decrease of ABA level in microtubers.

  14. In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia

    Ummi Nur Ain Abdul Razak

    2014-02-01

    Full Text Available Stevia rebaudiana Bertoni is a medicinal plants and commercially use as non-caloric sweetener for diabetic patient. In the present study, a protocol was developed for in vitro micropropagation using 6-benzylamino purine (BAP and Kinetin (Kn for the formation of multiple shoot proliferation and Indole-3-acetic acid (IAA, Indole-3-butyric acid (IBA and 1-Naphthaleneacetic acid (NAA for the induction of roots. Maximum shoot formation (7.82 ± 0.7 shoots per explants was observed on a Murashige and Skoog (MS medium supplemented with 0.5 mg L-1 BAP and 0.25 mg L-1 Kn. The maximum number of roots (30.12 ± 2.1 roots per explants was obtained on a MS medium containing 1.0 mg L-1 IBA. The well rooted plantlets were successfully weaned and acclimatized in plant soil with survival rate of 83.3 %.

  15. Effect of phytohormones on the growth of Scenedesmus quadricauda (Turp. Bréh

    Ewa Tatkowska

    2014-01-01

    Full Text Available Studies were made an the effect of indole-3-acetic acid (IAA, gibberellic acid (GA, kinetin (Kin, and various combinations of these substances upon dry matter increments, soluble protein content, and chlorophyll levels in the cultures of Scenedesmus quadricauda (Turp. Bréb. It was found that all these phytohormones, added separately to the medium, stimulated dry matter increment and protein content, although their effect depended on the concentration and the duration of culture. Addition of various combinations of the substances under study into the medium resulted in a less pronounced increment of dry matter, whereas the content of soluble protein significantly increased. The results reflect positive reaction of Scenedesmus quadricauda to the addition of phytohormones into the medium. They also suggest some differences in the action of particular hormones.

  16. In vitro technique for selection of radiation induced mutants of sweet potato

    Zhen, H.R.

    2001-01-01

    In vitro culture and irradiation techniques were used for obtaining mutants in sweet potato, lpomoea batatas. Callus was initiated from stem explants cultured on MS medium, supplemented with 1 mg NAA, 2 mg IAA, and 0.01 mg BA per litre. Calli were irradiated with 5 Gy gamma rays, and cultured on half-strength MS medium containing 2 mg IAA and 2 mg kinetin per litre. Variants for morphological changes in leaf and tuber skin and flesh color were found among the M 1 V 2 plants, derived from irradiated calli. A promising clone, 91-C3-15, with improved tuber shape and deep red skin was selected from the 776 M 1 V 2 population. (author)

  17. Cytokinins and urea derivatives stimulate seed germination in Lotus corniculatus L.

    Nikolić Radomirka

    2007-01-01

    Full Text Available We studied the effects of various cytokinins and urea derivatives on germination of aged seeds of in Lotus corniculatus L. The following substances were applied: N6-isoprenoid cytokinins (isopentenyl adenine and zeatin, adenine sulfate, N6-aromatic cytokinins (kinetin, benzyladenine and their N9-ribosides, N-benzyl-9-(2- tetrahydropyranyladenine, and urea derivatives (diphenylurea, thidiazuron, and chloro-pyridyl phenylurea. With the exception of adenine sulfate, all cytokinins increased the percentage of seed germination up to twofold, depending on their kind and concentration. It is concluded that cytokinins may be among the missing factors in aged seeds of L. corniculatus contributing to the implementation of their full germination potential. They could be used to improve germination of both freshly harvested and aged seed samples, if necessary. .

  18. Effects of growth regulators and activated charcoal for the micropropagation improvement of Dendrobium friedericksianum Rchb.f.

    Prasertsongskun, S. and

    2004-09-01

    Full Text Available The effects of different concentrations of growth regulators and activated charcoal on micropropagation were studied. Seedlings of orchid, Dendrobium friedericksianum Rchb.f, cultured on Vacin and Went medium containing 1.0 mg/l α-naphthaleneacetic acid (NAA and 1.0 mg/l kinetin (KN significantly yielded per cultured plant, the highest average number of leaves (4.40, roots (8.30, shoot length (1.67 centimeters and fresh weight (0.23 grams. Particularly when activated charcoal was added (0.3% w/v the average number of leaves, roots, shoot length and fresh weight per cultured plant increased. Percentage survival of the plantlet after 21 days of transplantation was 85%.

  19. Somatic embryogenesis and plant regeneration of Capsicum baccatum L.

    Peddaboina Venkataiah

    2016-06-01

    Full Text Available A plant regeneration protocol via somatic embryogenesis was achieved in cotyledon and leaf explants of Capsicum baccatum, when cultured on MS medium supplemented with various concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D, 0.5–5.0 mg l−1 in combination with Kinetin (Kn, 0.5 mg l−1 and 3% sucrose. Various stages were observed during the development of somatic embryos, including globular, heart, and torpedo-stages. Torpedo stage embryos were separated from the explants and subcultured on medium supplemented with various concentrations of different plant growth regulators for maturation. Maximum percentage (55% of somatic embryo germination and plantlet formation was found at 1.0 mg l−1 BA. Finally, about 68% of plantlets were successfully established under field conditions. The regenerated plants were morphologically normal, fertile and able to set viable seeds.

  20. Expression of the 1-SST and 1-FFT genes and consequent fructan accumulation in Agave tequilana and A. inaequidens is differentially induced by diverse (a)biotic-stress related elicitors.

    Suárez-González, Edgar Martín; López, Mercedes G; Délano-Frier, John P; Gómez-Leyva, Juan Florencio

    2014-02-15

    The expression of genes coding for sucrose:sucrose 1-fructosyltransferase (1-SST; EC 2.4.1.99) and fructan:fructan 1-fructosyltransferase (1-FFT; EC 2.4.1.100), both fructan biosynthesizing enzymes, characterization by TLC and HPAEC-PAD, as well as the quantification of the fructo-oligosaccharides (FOS) accumulating in response to the exogenous application of sucrose, kinetin (cytokinin) or other plant hormones associated with (a)biotic stress responses were determined in two Agave species grown in vitro, domesticated Agave tequilana var. azul and wild A. inaequidens. It was found that elicitors such as salicylic acid (SA), and jasmonic acid methyl ester (MeJA) had the strongest effect on fructo-oligosaccharide (FOS) accumulation. The exogenous application of 1mM SA induced a 36-fold accumulation of FOS of various degrees of polymerization (DP) in stems of A. tequilana. Other treatments, such as 50mM abscisic acid (ABA), 8% Sucrose (Suc), and 1.0 mg L(-1) kinetin (KIN) also led to a significant accumulation of low and high DP FOS in this species. Conversely, treatment with 200 μM MeJA, which was toxic to A. tequilana, induced an 85-fold accumulation of FOS in the stems of A. inaequidens. Significant FOS accumulation in this species also occurred in response to treatments with 1mM SA, 8% Suc, and 10% polyethylene glycol (PEG). Maximum yields of 13.6 and 8.9 mg FOS per g FW were obtained in stems of A. tequilana and A. inaequidens, respectively. FOS accumulation in the above treatments was tightly associated with increased expression levels of either the 1-FFT or the 1-SST gene in tissues of both Agave species. Copyright © 2013 Elsevier GmbH. All rights reserved.

  1. Effect of genotype, gelling agent, and auxin on the induction of somatic embryogenesis in sweet potato (Ipomoea batatas Lam.).

    El Abidine Triqui, Zine; Guédira, Abdelkarim; Chlyah, Averil; Chlyah, Hassane; Souvannavong, Vongthip; Haïcour, Robert; Sihachakr, Darasinh

    2008-03-01

    Lateral buds of six cultivars of sweet potato were induced to form embryogenic callus in a culture medium solidified with two types of gelling agents, Agar or Gelrite, and supplemented with various concentrations of auxins, 2,4-D, 2,4,5-T and Picloram. Of the six cultivars screened, only three gave an embryogenic response. Best results with an average of 3.53% embryogenic response were obtained with the medium solidified with Agar, while in Gelrite only 0.45% of lateral buds gave rise to embryogenic callus. The interaction between the genotype and auxins was highly significant; particularly the optimal response was obtained with cv. Zho and 865 yielding 10.7 and 14.7% somatic embryogenesis, respectively, in the medium containing 2,4,5-T or Picloram. The plant conversion was dramatically improved by subculture of the embryogenic callus on the medium with the combination of 1 microM 2,4-D and 1 microM Kinetin or 5 microM ABA alone before transfer of mature embryos onto hormone-free medium. The embryogenic callus of sweet potato and its sustained ability to further regenerate plants have regularly been maintained for several years by frequent subculture in 5 microM 2,4,5-T or the combination of 10 microM 2,4-D and 1 microM BAP or kinetin. The embryo-derived plants seemed apparently genetically stable and similar to the hexaploid parental plants, based on morphological analysis and their ploidy level determined by using flow cytometry.

  2. The Effect of Plant Growth Regulators and Different Explants on the Response of Tissue Culture and Cell Suspension Cultures of German Chamomile (Matricaria chamomilla L.

    L. Koohi,

    2014-07-01

    Full Text Available German chamomile (Matricaria chamomilla L. is one of the most important medicinal plants that its essential oils used in different medicinal industries. In this study which was carried out in 2013 growing season at the Faculty of Agricultural Sciences of the University of Mohaghegh Ardabili, the in vitro response of leaf and hypocotyl explants of German Chamomile in B5 medium supplemented with different levels of plant growth regulators including 2,4-D, naphthalene acetic acid (NAA, kinetin and 6-benzylaminopurine (BAP were investigated in a factorial experiment based on completely randomized design (CRD.In addition, cell suspension cultures were established and characterized. Hypocotyl and leaf explants exhibited cell proliferation and produced callus within 1-2 weeks. The highest fresh weight of the callus (264.1 mg was produced by leaf explants in the medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l BAP. However, the leaf explants cultured on medium containing 1.5 mg/l 2,4-D showed the lowest cell proliferation and callus yield (40.42 mg. The highest percentage of root induction from leaf explants (58.73% was observed on the medium containing 4 mg/l 2,4-D and 1 mg/l Kin, and from hypocotyl explants (48.61% was observed on medium supplemented with 1.5 mg/l NAA. The 42.22% of calli derived from hypocotyl explants on B5 medium supplemented with 4 mg/l NAA and 3 mg/l BAP, were friable. Cell suspension cultures of German chamomile were established by transferring of hypocotyl-derived friable calli into the MS medium supplemented with 1.5 mg/l 2,4-D and 1 mg/l kinetin. The growth curve of cell proliferations started 4 days after culture and continued to grow until day 13th, where the cells entered stationary phase.

  3. In vitro propagation and withaferin A production in Withania ashwagandha, a rare medicinal plant of India.

    Mir, Bilal Ahmad; Mir, Shabir Ahmad; Koul, Sushma

    2014-07-01

    Withania ashwagandha, belonging to the family Solanaceae, is an important medicinal herb of India with restricted geographic distribution. It is a rich source of withaferin A (WA) and other bioactive withanolides. In the present study a rapid in vitro mass propagation protocol of W. ashwagandha was developed from nodal explants. Nodal explants were cultured on MS medium supplemented with various concentrations and combinations of plant growth regulators (PGRs). The highest number of regenerated shoots per ex-plant (33 ± 2.7) and highest WA (13.4 ± 1.15 mg/g of DW) production was obtained on MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). In vitro raised shoots were further rooted on half-strength MS medium containing 2.0 μM Indole-3-butyric acid (IBA) and analyzed for WA production. The rooted plantlets when transferred to poly bags in the greenhouse showed 90 % survival frequency. Levels of WA were higher in the in vitro and ex vitro derived shoot and root tissues as compared to field grown mother plants. In an attempt to further maximize WA production, shoot cultures were further grown in liquid MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). Root cultures were grown on half strength MS liquid medium fortified with 2.0 μM of IBA. WA production in the liquid cultures was significantly higher compared to the static composition of the same media. This protocol, first of its kind in this plant, can be successfully employed for conservation, proliferation and large-scale production of WA. The regenerated plants can also be used in traditional medicine as an alternative to naturally collected plants.

  4. In vitro studies on callus induction in gamma-irradiated velvet bean seeds (Mucuna pruriens L.)

    Gupta, Ankit; Misra, Pragati; Shukla, Pradeep K.

    2014-01-01

    Gamma rays are often used for developing plants varieties that are agriculturally and economically important and have high productivity potential with the minimum input. Ionizing radiations are currently a very important way to create genetic variability that is not exists in nature or that is not available to the breeder. Irradiation treatments performed at in vitro culture has been also employed to increase genetic variability and mutants as a potential source of new commercial cultivars. Large number of research reports suggests also that mutagenesis in combination with tissue culture has high potential in plant breeding programs. Mucuna pruriens L., also known as velvet bean, contains L-DOPA, a precursor to the neurotransmitter dopamine and formulation of the seed power has been studied for management of Parkinson's disease. Seeds were exposed to different doses of gamma radiation (10 kGy, 20 kGy and 30 kGy) using 60 Co as source, at National Botanical Research Institute (NBRI) Lucknow. Gamma treated and untreated seeds (control) were inoculated in MS media supplemented with different phytohormone concentrations and combinations. The best callus induction was observed in control seeds on MS media supplemented with 0.5 mg/l kinetin, 2 mg/l NAA and 10 mg/l adenine sulphate, whereas gamma treated seeds showed poor callus induction in the same phytohormone concentrations. The callus induction was poor in control seeds on MS media supplemented with 1.0 mg/l kinetin, 2.0 mg/l NAA and 10 mg/l adenine sulphate, whereas gamma treated seeds showed even poor callus induction under the same phytohormone concentrations. The callus induction frequency was in declined gradually with the increasing dose of gamma radiation. Gamma treated seeds developed greenish and fragile callus and also showed decreased weight as compare to control which was white greenish, compact and heavier. (author)

  5. Tratamentos pré-germinativos em sementes de Didymopanax morototoni (Aubl. Dcne. et Planch

    Elci Terezinha Franco Henz

    2002-01-01

    Full Text Available Didymopanax morototoni (Aubl. Dcne. et Planch, (caixeta, is a native forest tree with economical importance in Brazil. It has a large natural distribution, with rapid growth and is a pioneer species. But the seeds of that species have a low rate of germination. The aim of this work was to evaluate the effect of the pre-treatments on the germination capacity. The following treatments were tested: immersion of seed in water for 8 h; chemical scarification with HCl and H2SO4 a 70%; washing with water and alcohol + water (1:1 at different times (15, 30 and 45 min. and mechanical scarification. In vitro germination tests were also carried out. Seeds without test were inoculated in the WPM medium solidified with agar (6 g/l in presence of kinetin and gibberelic acid on the concentration of 0; 0,1; 0,5; 1,0 mg/l, plus hydrolysed casein on the concentration of 250 mg/l. The incubation was at de 25 ±1°C, in germination chambers under photoperiod of 16 hours light under14, 3 μE.m-². s-¹ or in the darkness. The germination was improved by the pre-treatments rinsed in water or water plus alcohol in 45 minutes. The addition of kinetin and gibberelic acid allowed an improvement in the germination of 40 to 70%. The results confirmed that there was a partial dormancy due to teguments and point out to possible presence of inhibitory substances in them which can be removed by washing the seeds.

  6. Direct regeneration and efficient in vitro root development studies in lentil (lens culinaris medik)

    Sultana, T.; Majeed, N.; Naqvi, S.

    2016-01-01

    Lentil is a self-pollinating annual crop with increasing demand all over the world due to its high protein content and easy digestibility. However, like many other crops lentil too needs improvement for which conventional as well as biotechnological tools are to be employed. This study was aimed at development of tissue culture protocol especially targeting improved root development to ensure their establishment in soil in order to use their potential towards genetic manipulation. Two Pakistani lentil cultivars, Masoor-2002 and Manshera-89 were used to obtain cotyledonary nodes, epicotyl and hypocotyl explants. The explants were cultured on shoot regeneration medium containing different concentration of kinetin, BAP and tyrosine with the addition of GA3, with or without charcoal for shoot development. Masoor-2002, showed the highest frequency of shoot development on MS medium containing 5.5 mg/L tyrosine, 0.25 mg/L kinetin, 1.0 mg/L BAP, 0.1 mg/L GA3, using cotyledonary node as explant. The addition of 2 g/L of charcoal in shoot medium resulted in healthier plants, but the number of shoots were reduced. Regarding the effect of age of explants on regeneration frequency, cotyledonary nodes of age 4-6 days had higher regeneration potential. Well-developed shoots were shifted to rooting medium containing different concentration of auxin with or without charcoal. Healthier and more roots were observed on medium containing 4 mg/L IAA with addition of 2 g/L charcoal. Plants were better established (70% survival) in a soil mix containing perlite, vermiculite and peat moss in 1:1:1 ratio. (author)

  7. Influence of growth regulators on distribution of trichomes and the production of volatiles in micropropagated plants of Plectranthus ornatus

    Helna C. Passinho-Soares

    Full Text Available ABSTRACT The profile of volatile organic compounds, the glandular and non-glandular trichomes of Plectranthus ornatus, obtained by in vitro cultivation, was evaluated in plants grown in Murashide and Skoog medium supplemented with benylaminopurine at 4.5, 9.0, and 18.0 µM + naphthaleneacetic acid at 5.37 µM, kinetin at 4.7, 9.3 and 18.5 µM + naphthaleneacetic acid (5.37 µM or Murashide and Skoog 0 medium (as a control. Scanning Electron Microscopy was performed on samples of the third leaf node of the 90 days old plants obtained from treatment with 4.5 or 9.0 µM benylaminopurine, and 4.7 or 9.3 µM kinetin. Headspace Solid Phase Micro-Extraction of the 30, 60 and 90 days old in vitro plants permitted to determinate by GC/MS the composition comprised of 62 compounds. The data were analyzed using Principal Component Analysis and Hierarchical Clustering Analysis and, the major constituents of these oils after treatment and aging were monoterpenes and sesquiterpenes. Morphoanatomical analysis of trichomes, by Scanning Electron Microscopy, enabled the identification of non-glandular trichomes and four types of glandular trichomes, which comprised capitate and peltate glandular trichomes that were distributed on both sides of the leaf. We observed that the regulators influenced qualitative and quantitative profiles of the volatile organic compounds and the number and distribution of hairs on the leaf surface.

  8. REGENERASIRUMPUT LAUT Kappaphycus alvarezii (Doty MELALUI INDUKSI KALUS DAN EMBRIO DENGAN PENAMBAHAN HORMON PERANGSANG TUMBUH SECARA IN VITRO

    Emma Suryati

    2009-04-01

    Full Text Available Regenerasi rumput laut Kappaphycus alvarezii dilakukan dalam rangka penyediaan benih yang bermutu dan mempunyai keunggulan melalui induksi kalus dan embrio dengan penambahan hormon pertumbuhan yang diintroduksi ke dalam media kultur yang dapat memacu induksi kalus dan penebalan pigmen rumput laut. Media kultur yang digunakan adalah media Conwy padat dengan penambahan agar 0,8%-1,6%. Hormon perangsang tumbuh yang digunakan untuk memacu pertumbuhan kalus dan filamen embrio yaitu IAA (Indol acetic acid, kinetin, dan auxilin dengan konsentrasi berkisar 0,4-1 mg/L. Embrio yang dihasilkan merupakan anakan yang mempunyai sifat yang sama dengan induknya. Sintasan dan perkembangan embrio yang paling baik yaitu dengan penambahan IAA dengan konsentrasi 0,4 mg/L pada media padat. Pembentukan anakan dilakukan dengan mengiris embrio dan menumbuhkan pada media cair yang diperkaya dengan hormon yang sama. Pemeliharaan anakan pada media kultur dilakukan hingga mencapai ukuran 2-3 cm. Regeneration of seaweed Kappaphycus alvarezii was done to provide high quality seed through callus and embryo induction using plant growth regulator which was introducted to the culture medium. This growth regulator can stimulate the callus induction procces and thickening the seaweed pigment. Applied medium culture was agar medium with 0.8%-1.6% concentration enriched with Conwy and the applied growth regulators were IAA (Indol acetic acid, kinetin dan auxilin with 0.4-1 mg/L concentration range. Resulted embryo has the same characteristics with the stock. The best survival rate and embryo growth was IAA treatment with 0.4 mg/L concentration. Formation of embryo was done by transferring them from solid medium to the liquid one with the same growth regulator treatment. The nursery of the seed in culture medium was carried out until it has reached 2-3 cm in size.

  9. Indução de calos embriogênicos em explantes de cupuaçuzeiro Induction of embryogenics calli in cupuassu explants

    Maria das Graças Rodrigues Ferreira

    2004-08-01

    Full Text Available Objetivou-se a indução de calos embriogênicos em cupuaçuzeiro, em função do tipo de explante e meio de cultura. Foram testados como explantes, segmentos cotiledonares e eixos embrionários divididos em três partes: região da plúmula, radícula e hipocótilo. Os explantes foram cultivados em 2 diferentes meios de cultura: 1 MS suplementado com 2,4-D (1 mg L-1 e Cinetina (0,25 mg L-1; 2 MS acrescido de ANA (5 mg L-1 e Cinetina (0,25 mg L-1. Constatou-se que a região do hipocótilo foi a parte mais responsiva do eixo embrionário, formando calos com aspecto branco e friável. As auxinas testadas nos meios não estimularam o processo embriogênico em calos de cupuaçuzeiro.It was studied the induction of embryogenics calli in cupuassu, in function of kind of explant and culture medium. Cotyledons segments and embryonic axes were tested and divided in three parts: region of plumule, radicule and hypocotile. The explants were cultivated in two different culture medium: 1 MS supplemented with 2,4-D (1 mg L-1 and Kinetin (0,25 mg L-1; 2 MS supplemented with NAA (5 mg L-1 and Kinetin (0,25 mg L-1. The hypocotile region demonstrated to be more responsive segment of the embryonic axe, forming callus with white and friable aspect. No somatic embryogenesis was evidenced in callus of cupuassu with auxines testeds in the medium.

  10. Seed germination of medicinal plant, fennel (Foeniculum vulgare Mill), as affected by different priming techniques.

    Tahaei, Amirreza; Soleymani, Ali; Shams, Majid

    2016-09-01

    Reduced seed germination is among the most important factors adversely affecting crop stand and subsequent plant growth. Fennel (Foeniculum vulgare Mill) is an important medicinal plant with poor seed germination rate, occasionally. It is accordingly pertinent to find methods which can enhance fennel seed germination and remove the barriers of dormancy breaking. The present experiments studied the effects of two different priming (cold moist stratification and osmopriming) and 14 dormancy breaking techniques (hormonal, osmopriming, biopriming, chemical priming, and hydropriming) on the seed germination and seedling growth of two different fennel genotypes under growth chamber conditions. In the first and second experiment, the priming techniques including the time lengths of cold moist stratification (0, 15, 30, and 45 days) and the concentrations of polyethylene glycol 6000 (PEG6000, osmopriming at -0.99, -1.35, and -2.33 MPa) were used as the main plots. However, in both experiments, the dormancy breaking techniques and fennel genotypes were factorially combined and used as the subplots. Different seed- and seedling-related parameters including germination (%), plumule, radicle and seedling length, average germination time, rate and homogeneity of germination, and seed vigor index were determined. Both priming techniques were efficient on the enhancement of seed germination and seedling growth. Among the dormancy breaking techniques, Aminol Forte (biopriming), kadostim (biopriming), benzyl adenine + kinetin (biopriming), distilled water (hydropriming), gibberellin + kinetin (hormonal priming), and benzyl adenine + kinetin + gibberellin (biopriming) were the most effective ones. The related concentrations were equal to 100 mg/l, 10(-5) M, and 0.4 %. The fennel genotypes reacted significantly different under priming conditions. It is possible to enhance seed germination and seedling growth of fennel using priming and dormancy breaking

  11. Regeneration of three sweet potato (Ipomea batatas (L.)) accessions via meristem, Nodal and callus induction

    Addae-Frimpomaah, F.

    2012-11-01

    In vitro regeneration of three sweet potato accessions UE007, UK-BNARI and SA-BNARI using meristem, nodal cuttings or callus induction was studied. Meristematic explants cultured on Murashige and Skoog (1962) basal medium supplemented with low concentration of benzylaminopurine (BAP) or kinetin resulted in callus with or without shoot development which delayed shoot emergence. The degree of callus development increased as the concentration of the cytokinin in the culture medium increased. Although, callus development was comparatively lower on kinetin amended medium than BAP amended medium, Murashige and Skoog medium supplemented with 0.25mg/1BAP had the highest shoot induction (80%). For further differentiation of callus or shoots into distinct stem and leaves, the culture were transferred into fresh MS medium supplemented with 0.25mg/1 BAP, 0.1 mg/1 NAA and 0.1 mg/1 Gibberellic acid (GA 3 . To overcome the delay in shoot initiation using meristem culture, nodal cuttings of sweet potato were used as explants and cultured on MS medium amended with 0.3 - 0.9mg/1 BAP. All explants cultured on 0.3 or 0.6mg/1 BAP developed shoots. Furthermore, liquid MS medium amended with 0.25mg/1 BAP, 0.1mg/I NAA, and 0.1mg/1 GA 3 also enhanced early shoot development from nodal cutting explants compared to solid culture. Post flask acclimatisation of meristem or nodal cutting-derived plantlets showed that meristem derived plantlets were better acclimatised than nodal cutting plants due to vigorous root development leading to higher percentage survival in pots and subsequent tuber production. Callusogenesis was achieved when leaf lobe explants were cultured on CLC/ Ipomoea medium supplemented with 1.0 - 4.0mg/1 2,4-D with 4.0mg/1 2,4-D being the optimal concentration. However, the calli were non-embryogenic and therefore could not produce embryos when transferred to 0.1mg/1 BAP amended medium but rather produced either single or multiple shoots. The highest percentage shoot (83

  12. In-vitro regeneration of castor bean (ricinus communis, L) and physic nut (jatropha curcas, L.)

    Afful, N. T.

    2008-06-01

    Investigations were conducted on in vitro regeneration of physic nut (Jatropha curcas, 1.) and castor bean (Ricinus communis, 1.) using zygotic embryo culture, shoot tip and meristem culture as well as somatic embryogenesis. Zygotic embryos from different stages of fruit maturity cultured on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), kinetin and 2-isopentyl (2iP) produced vigorous seedlings with roots. However the optimal concentration of cytokinin for seedling development depended on the type cytokinin as well as the maturity stage of the fruit. Embryos from yellow fruits of Jatropha had the highest germination (100%) on an MS medium supplemented with 0.5 mg/L kinetin or 1 mg/L BAP while in Ricinus embryos from black fruits had the highest percentage germination (97%) on an MS medium modified with I mg/L 2iP or 0.5 or I mg/L BAP. Decrease in embryo viability especially in Jatropha as the seed matured may be due to high oil content in matured (black) fruit. Oil content in both Jatropha and Ricinus increased as the fruit matured from green to black and this may have effect on germination of zygotic embryos. In vitro culture response of shoot tip or meristem was significantly influenced by the concentration of growth regulators in the culture medium as well as the plant species. In both plant species, 2iP resulted in highest percentage shoot regeneration. The optimal concentration of 2iP was 0.5 mg/L for Ricinus shoot tip and 1.5 or 2.0 mg/L for Jatropha, indicating genotypic difference between the species. However excessive callus formation and browning in Ricinus led to the loss of all the regenerants. Shoot regeneration from meristem explants was also influenced by growth regulators with 2iP again being the best. Cultures of cotyledon and leaf lobe explants on MS medium supplemented with 2, 4-D and picloram resulted in callus induction with the exception of cotyledons obtained from black fruits. However transfer of calli onto fresh

  13. EFFECTS OF SOME PLANT GROWTH REGULATORS ON JASMONIC ACID INDUCED INHIBITION OF SEED GERMINATION AND SEEDLING GROWTH OF BARLEY

    Kürşat ÇAVUŞOĞLU

    2009-02-01

    Full Text Available Abstract: The effects of gibberellic acid, kinetin, benzyladenine, ethylene, 24-epibrassinolide and polyamines (spermine, spermidine, putrescine, cadaverine on jasmonic acid inhibition of seed germination and seedling growth of barley were studied. All of the plant growth regulators studied were determined to have a succesful performance in reversing of the inhibitory effects of jasmonic acid on the seed germination and seedling growth. Moreover, the above mentioned growth regulators overcame the inhibitory effect of JA on the percentages of germination and coleoptile emergence in the same ratio, while GA3 was the most successful hormone on the fresh weight and radicle and coleoptile elongation in comparison with the other growth regulators. Key words: Barley, jasmonic acid, plant growth regulator, seed germination, seedling growth ARPANIN TOHUM ÇİMLENMESİ VE FİDE BÜYÜMESİNİN JASMONİK ASİT TEŞVİKLİ İNHİBİSYONU ÜZERİNE BAZI BİTKİ BÜYÜME DÜZENLEYİCİLERİNİN ETKİLERİ Özet: Arpanın tohum çimlenmesi ve fide büyümesinin jasmonik asit inhibisyonu üzerine gibberellik asit, kinetin, benziladenin, etilen, 24-epibrassinolit ve poliaminlerin (spermin, spermidin, putressin, kadaverin etkileri araştırılmıştır. Çalışılan bitki büyüme düzenleyicilerinin tümünün tohum çimlenmesi ve fide büyümesi üzerinde jasmonik asitin engelleyici etkisini tersine çevirmede başarılı bir performansa sahip oldukları belirlenmiştir. Dahası, yukarıda sözü edilen büyüme düzenleyicileri çimlenme ve koleoptil çıkış yüzdeleri üzerinde aynı oranda etkili olurken, taze ağırlık ve radikula ve koleoptil uzaması üzerinde diğer büyüme düzenleyicileri ile karşılaştırıldığında en başarılı hormon GA3 olmuştur. Anahtar kelimeler: Arpa, jasmonik asit, bitki büyüme düzenleyicisi, tohum çimlenmesi, fide büyümesi

  14. Protocol for in vitro somatic embryogenesis and regeneration of rice (Oryza sativa L.).

    Verma, Dipti; Joshi, Rohit; Shukla, Alok; Kumar, Pramod

    2011-12-01

    Development of highly efficient and reproducible plant regeneration system has tremendous potential to provide improved technology to assist in genetic transformation of indica rice cultivars for their further exploitation in selection. For the development of a highly reproducible regeneration system through somatic embryogenesis, mature embryos of highly popular rice cultivars i.e., Govind (for rainfed areas), Pusa Basmati-1 (aromatic basmati) and Jaya (for irrigated areas) were used. Optimum callus formation (%) to MS medium supplemented with 2, 4-D was obtained at 12.0 microM in Govind, 14.0 microM in Jaya and 15.0 microM in Pusa Basmati-1. All the cultivars showed good proliferation on MS medium without hormone. In Govind, highest embryogenic response was observed in MS medium supplemented with 2, 4-D (0.4 microM) + kinetin (0.4 microM), while in Pusa Basmati-1 with 2, 4-D (0.4 microM) + kinetin (2.0 microM) and in Jaya on hormone-free MS medium. Excellent embryo regeneration in Govind was observed on MS medium supplemented with low concentrations (1.1 microM) of BAP or hormone-free MS medium, while in Pusa Basmati-1 and Jaya embryogenesis was observed on MS medium supplemented with higher concentration of BAP (2.2 microM). Similarly, maximum plantlets with proliferated roots were observed in Govind on hormone-free MS medium, while in Pusa Basmati-1 and Jaya on MS medium supplemented with high concentration of NAA (4.0 microM). Developed plantlets were further successfully acclimatized and grown under pot culture up to maturity. Further the yield potential of in vitro developed plants was accessed at par to the direct seeded one under pot culture. Present, protocol standardizes somatic embryogenesis and efficient regeneration of agronomically important, high yielding and diverse indica rice cultivars which can be utilized as an efficient tool for molecular studies and genetic transformation in future.

  15. ISOLASI DAN KULTUR PROTOPLAS RUMPUT LAUT Kappaphycus alvarezii DI LABORATORIUM

    Emma Suryati

    2007-12-01

    Full Text Available Isolasi protoplas rumput laut K. alvarezii, telah dilakukan dalam rangka penyiapan protoplas untuk penyilangan melalui fusi protoplas. Metode yang digunakan antara lain melalui cara kimia dengan melisis tallus rumput laut dengan campuran enzim komersial, kemudian enzim yang berasal dari viscera keong mas baik yang segar maupun yang beku, dengan media kultur yang digunakan pada pemeliharaan makro algae antara lain Conwy, PES, dan air laut steril. Tallus rumput laut yang digunakan berasal dari bagian pangkal, tengah dan ujung. Protoplas yang hidup diuji menggunakan evans blue 0,1%, hormon perangsang tumbuh yang digunakan pada media pertumbuhan antara lain auxin, IAA, dan Kinetin. Pengamatan dilakukan terhadap jumlah protoplas hidup, pertumbuhan, dan sintasan.  Hasil percobaan memperlihatkan bahwa enzim yang paling baik digunakan adalah campuran enzim komersial dengan media kultur Conwy dengan jumlah protoplas mencapai 19,8 x 106 sel/mL, bagian tallus yang paling baik adalah bagian pangkal berkisar antara 8,1x106 hingga 18,8 x 106 sel/ mL.  Perangsang tumbuh yang paling baik adalah auxin. Filamen terbentuk setelah 5 hari dengan fotoperiod L:D=12:12. Isolation of seaweed’s protoplast Kappaphycus alvarezii had been done to provide protoplast for crossbreeding purpose by protoplast fusion. The method was chemically done by lyses of tallus used commercial enzyme mixture, enzyme from viscera of snail both fresh and frozen, culture media were Conwy (CW, PES, and sterile sea water (SSW which were used to maintain the macro algae. Part of used tallus were upper, middle and tip of tallus. The viable protoplast was examined by using 0.1% evans blue and the growth-stimulating hormone were auxin, IAA, and Kinetin. Observation was concerned to the amount of viable protoplast, the growth, and the long live. Result showed that the best enzyme was commercial enzyme mixture with Conwy as the best culture media, provided protoplast until 19.8 x 106 cell/mL. The

  16. Picroside I and Picroside II from Tissue Cultures of Picrorhiza kurroa

    Ganeshkumar, Yamjala; Ramarao, Ajmera; Veeresham, Ciddi

    2017-01-01

    Background: Picrorhiza kurroa (PK) belongs to Scrophulariaceae family and is a representative endemic, medicinal herb, widely distributed throughout the higher altitudes of alpine Himalayas from west to east, between 3000 and 4500 m above mean sea level. Objective: The objective of the present study is to assess the production of picroside I and picroside II from tissue cultures of PK. Materials and Methods: Auxiliary shoot tips of PK were incubated in Murashige and Skoog medium supplemented with indole-3-butyric acid and kinetin phytohormones. The callus produced was collected at different time intervals and was processed for extraction of picroside I and picroside II followed by thin layer chromatography and high-performance liquid chromatography HPLC analysis. Results: The maximum growth index was found to be 5.109 ± 0.159 at 16-week-old callus culture. The estimation of picroside-I and picroside-II was carried out by (HPLC) analysis; quantity of secondary metabolite found to be 16.37 ± 0.0007 mg/g for PK-I and 6.34 ± 0.0012 mg/g for PK-II. Conclusion: This is the first attempt to produce the Picroside-I and II in large amount by the tissue culture technique. It can be observed that the method of callus culture can be used in production of secondary metabolites Picroside-I and II from PK SUMMARY Picrorhiza kurroa is a high value medicinal herb due to rich source of hepatoprotective metabolites, Picroside-I and Picroside-II. The medicinal importance of P. kurroa is due to its pharmacological properties like hepatoprotective, antioxidant (particularly in liver), antiallergic and antiasthamatic, anticancer activity particularly in liver and immunomodulatory. Shoot apices which were produced a good response was inoculated on selected medium i.e., on MS medium containing 2, 4 D (mg/l) + KN (1mg/l) for induction of callus. The initiation of callus was observed after 4weeks and it was light green and fragile Maximum growth was observed with 3% w/v of sucrose

  17. The landscape of cytokinin binding by a plant nodulin

    Ruszkowski, M.; Szpotkowski, K.; Sikorski, M. [Polish Academy of Sciences, Poznan (Poland); Jaskolski, M., E-mail: mariuszj@amu.edu.pl [Polish Academy of Sciences, Poznan (Poland); A. Mickiewicz University, Poznan (Poland)

    2013-12-01

    The crystal structures of complexes of M. truncatula nodulin 13 with four cytokinins, trans-zeatin, N{sup 6}-isopentenyladenine, kinetin and N{sup 6}-benzyladenine, show an unusual mode of dimerization of this PR-10-fold plant protein. The cytokinin-binding mode in the internal cavity of the protein is the same in each complex and resembles the pattern found in the cytokinin receptor protein. Nodulation is an extraordinary symbiotic interaction between leguminous plants and nitrogen-fixing bacteria (rhizobia) that assimilate atmospheric nitrogen (in root nodules) and convert it into compounds suitable for the plant host. A class of plant hormones called cytokinins are involved in the nodulation process. In the model legume Medicago truncatula, nodulin 13 (MtN13), which belongs to the pathogenesis-related proteins of class 10 (PR-10), is expressed in the outer cortex of the nodules. In general, PR-10 proteins are small and monomeric and have a characteristic fold with an internal hydrophobic cavity formed between a seven-stranded antiparallel β-sheet and a C-terminal α-helix. Previously, some PR-10 proteins not related to nodulation were found to bind cytokinins such as trans-zeatin. Here, four crystal structures of the MtN13 protein are reported in complexes with several cytokinins, namely trans-zeatin, N{sup 6}-isopentenyladenine, kinetin and N{sup 6}-benzyladenine. All four phytohormones are bound in the hydrophobic cavity in the same manner and have excellent definition in the electron-density maps. The binding of the cytokinins appears to be strong and specific and is reinforced by several hydrogen bonds. Although the binding stoichiometry is 1:1, the complex is actually dimeric, with a cytokinin molecule bound in each subunit. The ligand-binding site in each cavity is formed with the participation of a loop element from the other subunit, which plugs the only entrance to the cavity. Interestingly, a homodimer of MtN13 is also formed in solution, as confirmed

  18. High Frequency Multiple Shoot Induction of Curculigo orchioides Gaertn.: Shoot Tip V/S Rhizome Disc

    K. S. Nagesh

    2008-09-01

    Full Text Available Curculigo orchioides Gaertn. is an endangered medicinal plant with anticancer properties. The rhizome and tuberous roots of the plant have been used extensively in India in indigenous medicine. Due to its multiple uses, the demand for Curculigo orchioides is constantly on the rise; however, the supply is rather erratic and inadequate. Destructive harvesting, combined with habitat destruction in the form of deforestation has aggravated the problem. The plant is now considered ‘endangered’ in its natural habitat. Therefore, the need for conservation of this plant is crucial. Here, we describe a successful protocol for multiple shoot induction of C. orchioides using shoot tip and rhizome disc. We find that proximal rhizome discs are optimal for high frequency shoot bud formation than shoot tip and distal rhizome disc. We observed a synergistic effect between 6-benzylaminopurine (BAP and kinetin (KN (each at 1 mg/L on the regeneration of shoot buds from proximal rhizome disc than shoot tip explant. Optimum root induction was achieved on half-strength MS liquid medium supplemented with 1 mg/L of indole-3-butyric acid (IBA. The in vitro raised plantlets were acclimatized in green house and successfully transplanted to natural condition with 90% survival.

  19. fektivitas Penambahan Elisitor Asam Jasmonik dalam Peningkatan Sintesis Senyawa Bioaktif Andrografolid pada Kultur Suspensi Sel Sambiloto (Effectiveness of Jasmonic Acid Elicitor Addition for Andrographolide Synthesis Induction of Sambiloto Culture

    Noor Aini Habibah

    2009-03-01

    Full Text Available In this research, we have studied synthesis of improvement of andrographolid bioactive compound on cell culture of sambiloto by addition of jasmonic acid. The essential problems in this research are firstly, the effects of addition of jasmonic acid either can induce or not andrographolide synthesis improvement of cell culture of sambiloto and secondly, to observe the largest content of andrographolide in jasmonic acid concentrations. Meanwhile, the purpose of this research are to observe the functions of jasmonic acid elicitor for induction of andrographolide synthesis improvement of cell culture of sambiloto and to optimize jasmonic acid concentrations which can produce the largest andrographolide content. The independent variable is concentration of addition of jasmonic acid on cell culture and the dependent variable are the growth of cell suspension culture and andrographolide bioactive content. Experiment result show that the optimum medium of sambiloto cell consist of Murashige & Skoog (1962 medium supplemented by 0,5 ppm kinetin and 2,4-D 5 ppm. The cell growth phases are the followings : lag phase at age of 0-5 days, exponential phase of 5-15 days, and stationary phase at age of longer than 15 days. The highest andrographolide was 4,66 x 10-2 reached in cell culture was supplemented with 10 µM jasmonic acid. Keywords : andrographolide, sambiloto cell suspension culture, jasmonic acid elicitor.

  20. In vitro regeneration of Drosera burmannii Vahl.: a carnivorous plant of north-east India.

    Yanthan, J Sureni; Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

    2017-06-01

    An efficient in vitro regeneration protocol has been developed from shoot tips of Drosera burmannii Vahl., a carnivorous plant of north-east India. Various plant growth regulators were used to study their efficacy in the induction of multiple shoots and roots. Of the various treatments, the maximum number of shoots (28.8 ± 1.5) and roots (9.7 ± 0.6) was observed in one-fourth strength standard medium (MS with 50 mg/l citric acid and 10 mg/l ascorbic acid) supplemented with 4 mg/l 6-benzylaminopurine (BAP) and 4 mg/l α-naphthalene acetic acid (NAA) followed by 26.8 ± 1.4 shoots in one-fourth strength SM fortified with 4 mg/l kinetin (KN) and 4 mg/l NAA. The well-developed plantlets with shoots and roots were potted in small plastic glasses filled with a mixture of sand and farmyard manure (3:1); these plantlets when transferred to a glasshouse for hardening and acclimatization showed 90% survival.

  1. Effect of the genotype and gamma irradiation on the anther cultures of a 10x10 diallel cross of wheat

    Molina, L.; Aldana, F.

    2001-01-01

    Anther culture responsiveness, irradiation effect and reciprocal effect were evaluated on ten genotypes (V1-V10) and a 101x0 diallel cross. Gamma irradiation dose of 100 Gy was applied to seeds of parents and F 1 cross from which the donor plants were grown. Non-irradiated donor plants were also used for comparison. Anthers were plated on potato-2 callus induction medium and calli formed were transferred to MS medium supplemented with sucrose (3%), indolacetic acid (1.0 mg/L), kinetin (1.0 mg/L), inositol (100 mg/L) and solidified with agar (0.7%). Genotypes showed big differences for callus induction, plant regeneration and anther culturability rate. The most responsive materials were V2, V10 and V5 with 76.0, 27.4 and 10.8 green plants per 100 anthers respectively. No irradiation effect was found for the parents nor the F 1 crosses on the pooled data. Mean anther culture response of specific genotypes showed that irradiation significantly increased anther culturability rate of V3 from 0.1 to 27.6 green plants per 100 anthers. No reciprocal effect was observed. (author)

  2. MICROPROPAGATION OF ADULT TREE OF PTEROCARPUS MARSUPIUM ROXB. USING NODAL EXPLANTS

    Shipra JAISWAL

    2015-12-01

    Full Text Available Attempts were made for in vitro propagation of Pterocarpus marsupium Roxb., belonging to family Fabaceae, an economically important multipurpose tree. The tree is scared with noval antidiabetic properties. The tree shows poor seed germination capacity (30% due to hard seed coat and conventional vegetative regeneration methods are a complete failure. Therefore, the propagation of this tree by tissue culture techniques is an urgent need and well justified. Nodal segments containing axillary bud from 10 years old tree of P. marsupium were evaluated for axillary shoot proliferation on Murashige and Skoog’s (MS basal medium fortified with BAP (6–benzylaminopurine and kinetin (Kn singly or in combinations with auxins at different concentrations. The best shoot proliferation was obtained with 13.95 µM Kn + additives (568 µM Ascorbic acid, 260 µM Citric acid, 605 µM Ammonium sulphate and 217 µM Adenine sulphate in MS medium where 64.44% of the axillary buds responded with development of (2.51±0.10 shoots. Multiplication of in vitro shoots were achieved on MS Medium supplemented with Kn (9.30 µM + NAA (0.54 µM and additives. Half strength MS medium supplemented with 4.92 µM IBA induced in vitro rooting of in vitro shoots. In vitro regenerated plantlets with well developed roots were successfully hardened in a greenhouse.

  3. Detection of irradiated mushrooms by GC/MS analysis of lipid-derived hydrocarbons

    Delincee, H.; Koller, W.D.

    1993-01-01

    A number of methods has been developed for the detection of irradiated foods in recent years, and in the case of mushrooms several methods have been proposed, of which the thermoluminescence (TL) measurements seem to be the most valuable. However, in several cases mineral contamination of fresh mushrooms is so extremely low that not enough minerals can be isolated for TL analysis. In that case an alternative method is needed to detect the radiation treatment of mushrooms. Several methods including TTC (2,3,5-triphenyl-tetrazolium-chloride) staining, kinetin treatment, dropping out of spores and mirco-gel electrophoresis of spores, were tested, but the most promising method was the GC/MS analysis of radiation-induced lipid-derived hydrocarbons in spite of the low fat content - around 0.2-0.3% - of mushrooms. Successful results were achieved by GC/MS analysis of the radiolytic hydrocarbons. Although mushrooms have a low fat content, by extracting a large quantity, in this case 500 g of mushrooms, about 1.2-1.5 g of fat could be obtained. The main fatty acids of mushroom fat and some of their expected cleavage products on irradiation - the c n-1 hydrocarbon which has one C atom less than the parent fatty acid and the C n-2:1 hydrocarbon, which has two C atoms less and an additional double bond in position 1 - are given. (orig./Vhe)

  4. Differentiation of shoot elements from the rachis of Secale cereale L.

    Jan J. Rybczyński

    2014-01-01

    Full Text Available In vitro culture of young Secale cereale spikes is described with special attention given to changes in development of the rachis. 7 mm explants were cultivated on a modified M u r a s h i g e and S k o o g (1962 medium (MS supplemented with 2,4-D (dichlorophenoxyacetic acid, NAA (α-naph-thaleneacetic acid, IAA (β-indoleacetic acid, 2,4,5-T (trichlorophenoxyacetic acid, and ZEA (zeatin, KIN (kinetin, BAP (6-benzylaminopurine, IPA (izopentenyladenine or 2 iP in numerous combinations and concentrations. Rachises differentiated branches with node construction in the presence of synthetic auxin. Rhizogenesis of explants was stimulated by 2,4-D and 2,4,5-T. 2,4,5-T + IPA appeared to be the best combination for callus regeneration. Many meristematic centres were found previously and combinations of NAA + KIN stimulated direct, whereas 2,4-D + ZEA indirect, differentiation of shoot elements. Development was observed from plantlets to flowering plants.

  5. Embriogênese somática do caquizeiro Somatic embryogenesis of japanese persimmon

    Dayse Cristina de Carvalho

    2004-08-01

    through somatic embryogenesis. Zygotic embryos excised from fruits collected from adult plants were tested at several developmental phases. They were collected during 22 weeks after 4 weeks blossoming. The basic medium tested was MS½NO3. Initial induction medium was supplemented with 20 µM 2,4-D + 2 µM kinetin. Dark calluses obtained were transferred to induction medium, with concentrations of 10 and 20 µM 2,4-D + 2 µM kinetin. The calli with pro-embryogenic masses were transferred to a maintenance and multiplication medium, with 2 µM kinetin and 2,4-D at the concentrations of 2.5, 5.0 and 10 µM. Embryogenic masses formed were transferred to a maturation medium supplemented with 0.5 µM IBA and 5, 10 and 20 µM 2iP. Embryos formed were isolated in two conversion media, one with 5 µM 2-iP + 5 µM GA3 and 0.5 µM IBA, and another medium with 0.5 µM GA3 and BAP at 0, 0.25, 0.5 and 1.0 µM. Indirect somatic embryogenesis were obtained from mature zygotic embryos collected after 22 weeks, when cultivated in culture medium with 10 µM 2,4D combined with 2 µM kinetin. Embryos maintenance and multiplication were more efficient with 5 µM 2,4-D, in which the pro-embryos advanced to the globular embryos phase. At the maturation phase the concentrations of 2-iP tested promoted globular embryos to more advanced stages of ontogeny, such as cordiform, torpedo and cotiledonary. Supplementation of the culture medium with 1 µM BAP generated better developed plants, with highest number of leaves and size.

  6. Organogênese direta e aclimatização de plantas de patchouli Direct organogenesis and acclimatization of patchouli

    Maria de Fátima Arrigoni-Blank

    2011-06-01

    aromatic species whose essential oil is largely employed by perfume industry. This conventional propagation is carried out using cuttings. Micropropagation is an alternative for clonal propagation of pathogens free individuals in large scale. We analyzed the influence of different combinations of auxins and kinetin in patchouli organogenesis and different kinds of substrate mixtures for the acclimatization of micropropagated plantlets. Two tests of organogenesis induction were carried out. In the first, five kinetin concentrations (0.0; 1.0; 2.0; 4.0 and 6.0 mg L‑1 and four IAA concentrations (0.0; 0.5; 1.0 and 2.0 mg L-1 were tested, and in the second, five kinetin concentrations (0.0; 0.5; 1.0; 2.0 and 4.0 mg L-1 and three NAA concentrations (0.0; 0.1 and 0.5 mg L-1. For acclimatization the following substrates were analyzed: coconut dust + formulated fertilizer (NPK 3-12-6 + Ca, S, Zn, B, Cu, Fe, Mn and B at 3.0; 2.5; 0.1; 0.025; 0.01; 0.075; 0.05 and 0.0015%, respectively at the concentration of 12 g L-1 + limestone (1 g L-1 [PBC]; coconut dust + vermiculite (2:1 + formulated fertilizer at the concentration of 12 g L-1 + limestone (1 g L-1 [PBCV (2:1]; coconut dust + vermiculite (1:1 v/v + formulated fertilizer at the concentration of 12 g L-1 + limestone (1 g L-1 [PBCV (1:1]; coconut dust + limestone (1 g L-1 + MS salts [PCMS]; vermiculite added with MS salts, using 15 mL of the salt mixture per plant [VMS]; vermiculite + formulated fertilizer at the concentration of 12 g L-1 [VMB]. Direct organogenesis can be promoted by MS medium supplemented with 2.47 mg L-1 of kinetin and 0.1 mg L-1 of NAA. The shoot elongation, individualization and shoot rooting steps, however, were promoted in MS medium without growth regulators. For acclimatization one of the best results was obtained using PBCV (1:1.

  7. Micropropagation of bioencapsulation and ultrastructural features of sainfoin (Onobrychis viciifolia) grown in vivo and in vitro.

    Mohajer, Sadegh; Mat Taha, Rosna; Mohajer, Minoo; Khorasani Esmaeili, Arash

    2014-01-01

    To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0-2.0 mg L(-1)) and Indole-3-acetic acid (0-2.0 mg L(-1)) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1:1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) in in vivo and 0.121 (abaxial), 0.201 (adaxial) in in vitro leaves.

  8. Micropropagation and assessment of genetic fidelity of Henckelia incana: an endemic and medicinal Gesneriad of South India.

    Prameela, J; Ramakrishnaiah, H; Krishna, V; Deepalakshmi, A P; Naveen Kumar, N; Radhika, R N

    2015-07-01

    Henckelia incana is an endemic medicinal plant used for the treatment of fever and skin allergy. In the present study shoot regeneration was evaluated on Murashige and Skoog's (MS) medium supplemented with auxins, Indole-3-acetic acid (IAA), Indole-3- butyric acid (IBA), 1-Naphthaleneacetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and cytokinins, 6-Benzylaminopurine (BAP) and Kinetin (Kn) at concentrations of 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 mgl(-1). MS medium with IBA (18.08), NAA (17.83) and IAA (17.58) at 0.5 mgl(-1) concentrations showed efficient regeneration. Regenerated shoots were rooted on half-strength MS medium with and without 0.5 mgl(-1) IBA or NAA. The plantlets were successfully hardened in rooting trays (peat, vermiculite and sand) and transferred to field mileu. The genetic fidelity of in vitro raised plants was assessed by using three different single primer amplification reaction (SPAR) markers namely random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and direct amplification of mini-satellite DNA region (DAMD). The results consistently demonstrated true-to-true type propagation. This is the first report of in vitro propagation and establishment of true-to-true type genetic fidelity in H. incana.

  9. Micropropagation of nucellar embryos and their histological comparative study for regeneration ability with other explants of kinnow mandarin (Citrus reticulata Blanco)

    Kazmi, S.K.; Khan, S.; Kabir, N.

    2018-01-01

    Polyembryony is the most beneficial and distinct character in citrus seeds. This characteristic can be beneficial in citrus improvement programs. Nucellar embryos developed from nucellar wall in citrus seeds along with zygotic embryo are found to have high plants regeneration ability in comparison to zygotic embryos. Under In vitro culture conditions, nucellar embryos were detected and multiplied on MT media with 0.5ml of coconut water along with 50g/L sucrose. These nucellar embryonic tissues were also regenerated on MT media upplemented with 0.5 mg/L BAP and 0.5 mg/L kinetin. Many shoots were regenerated from nucellar embryonic tissue; these shoots were rooted on MT media supplemented with IBA 0.5mg/L and NAA 0.1 mg/L in addition to activated charcoal 0.5gm/L. The resulting plantlets were acclimatized in the green house. After micropropagation, histological studies of nucellar embryonic tissues were carried out under fluorescence microscope to examine their high regeneration ability in comparison with usual plant parts, like seeds, shoots as well as with tissue culture stages including embryogenic and non-embryogenic callus. It was found that nucellar embryos have more regeneration ability as compared to usual plant parts and other tissue culture stages. (author)

  10. An effective protocol for micropropagation of edible bamboo species (Bambusa tulda and Melocanna baccifera) through nodal culture.

    Waikhom, Sayanika Devi; Louis, Bengyella

    2014-01-01

    High demand for edible bamboo shoots of Bambusa tulda and Melocanna baccifera in many Asian ethnic groups has led to the need for developing intensive bamboo farming. To achieve this, in vitro regeneration of bamboo plantlets is needed due to the long and irregular bamboo flowering cycle and scarcity of bamboo seeds. An effective protocol for plantlets regeneration in B. tulda and M. baccifera from nodal explants following validation of the species using the sequence of trnL-F intergenic spacer region is described. Effective axillary bud breaking was achieved at 3 mg/L of 6-benzylaminopurine (BAP) in MS medium. Importantly, combining 2 mg/L of kinetin (Kn) with 3 mg/L of BAP produced a synergistic effect for shoot multiplication in B. tulda and M. baccifera. Under optimized conditions in half-strength MS medium supplemented with 3 mg/L of indole-3-butyric acid (IBA), 10 mg/L of coumarin, and 3% sucrose, profuse production of dark-brown rhizome in B. tulda and abundant rooting (81.67%, P < 0.05, F = 15.46) for M. baccifera within 30 days were achieved. The established protocol and the validation of the reported species at the molecular level will be of help to stakeholders in edible bamboo trade to conserve gene-pool and increase productivity.

  11. An Improved Micropropagation Protocol by Ex Vitro Rooting of Passiflora edulis Sims. f. flavicarpa Deg. through Nodal Segment Culture.

    Shekhawat, Mahipal S; Manokari, M; Ravindran, C P

    2015-01-01

    A procedure for rapid clonal propagation of Passiflora edulis Sims. f. flavicarpa Deg. (Passifloraceae) has been developed in this study. Nodal explants were sterilized with 0.1% HgCl2 and inoculated on Murashige and Skoog (MS) basal medium. The addition of 2.0 mgL(-1) 6-benzylaminopurine (BAP) to MS medium caused an extensive proliferation of multiple shoots (8.21 ± 1.13) primordial from the nodal meristems. Subculturing of these multiple shoots on the MS medium augmented with 1.0 mgL(-1) of each BAP and Kinetin (Kin) was successful for the multiplication of the shoots in vitro with maximum numbers of shoots (25.73 ± 0.06) within four weeks of incubation. Shoots were rooted best (7.13 ± 0.56 roots/shoots) on half strength MS medium supplemented with 2.0 mgL(-1) indole-3 butyric acid (IBA). All in vitro regenerated shoots were rooted by ex vitro method, and this has achieved 6-7 roots per shoot by pulsing of cut ends of the shoots using 200 as well as 300 mgL(-1) IBA. The plantlets were hardened in the greenhouse for 4-5 weeks. The hardened plantlets were shifted to manure containing nursery polybags after five weeks and then transferred to a sand bed for another four weeks for acclimatization before field planting with 88% survival rate.

  12. Micropropagation of Asparagus by in vitro shoot culture.

    Stajner, Nataša

    2013-01-01

    Asparagus officinalis is most extensively studied species within the genus Asparagus, which is well known as garden asparagus. This species is dioecious with unisexual flowers, which means that generative propagation gives roughly equal number of male and female plants. Male plants are high yielders and preferred commercially over female plants. Tissue culture techniques could efficiently promote vegetative propagation of male plants and pave the way for efficient plant breeding.This chapter describes an efficient micropropagation protocol for developing rapid growing in vitro Asparagus shoot cultures. The source of explants, inoculation, and shoot proliferation, followed by shoot propagation, rooting, and acclimatization is described. The optimal medium for Asparagus micropropagation described in this chapter is composed of MS macro- and microelements and a combination of auxins and cytokinins. Plant growth regulators NAA, kinetin, and BA were used in various concentrations. Three different media representing the whole micropropagation protocol of Asparagus are described; medium for shoot initiation, medium for shoot multiplication, and medium for root formation. By in vitro propagation of Asparagus, root initiation is difficult, but can be promoted by adding growth retardant ancymidol which also greatly promotes shoot development and suppresses callus formation.

  13. Factors affecting in vitro plant regeneration of the critically endangered Mediterranean knapweed ( Centaurea tchihatcheffii Fisch et. Mey)

    Ozel, Cigdem Alev; Khawar, Khalid Mahmood; Mirici, Semra; Ozcan, Sebahattin; Arslan, Orhan

    2006-10-01

    Habitat destruction has resulted in the extinction of many plant species from the earth, and many more face extinction. Likely, the annual endemic Mediterranean knapweed ( Centaurea tchihatcheffii) growing in the Golbasi district of Ankara, Turkey is facing extinction and needs urgent conservation. Plant tissue culture, a potentially useful technique for ex situ multiplication, was used for the restoration of this ill-fated plant through seed germination, micropropagation from stem nodes, and adventitious shoot regeneration from immature zygotic embryos. The seeds were highly dormant and very difficult to germinate. No results were obtained from the micropropagation of stem nodes. However, immature zygotic embryos showed the highest adventitious shoot regeneration on Murashige and Skoog (MS) medium, containing 1 mg l-1 kinetin and 0.25 mg l-1 NAA. Regenerated shoots were best rooted on MS medium containing 1 mg l-1 IBA and transferred to the greenhouse for flowering and seed set. As such, the present work is the first record of in vitro propagation of critically endangered C. tchihatcheffii, using immature zygotic embryos, and is a step forward towards conservation of this indigenous species.

  14. Micropropagation of Gerbera (Gerbera jamesonii Bolus).

    Minerva, Ghani; Kumar, Surinder

    2013-01-01

    Gerbera (Gerbera jamesonii Bolus) is one of the most popular ornamental flowers worldwide and used both as cut flower and potted plant. Some of them show excellent agronomic characters such as color, floral diameter, stem length, and vigor, which make this plant of commercial importance. Conventionally, multiplication is done through seeds or rhizome cuttings. Rapid multiplication of elite cultivars of Gerbera, with improved agronomic traits, has been achieved by using both direct and indirect tissue culture methods. Direct shoot regeneration was accomplished from stem apices on MS medium supplemented with 1 mg/L 6-benzyladenine (BA) and 1 mg/L kinetin. Indirect shoot induction succeeded from callus differentiation has been achieved on MS medium containing 2 mg/L 2,4-dichlorophenoxyacetic acid, 0.5 mg/L indole-3-acetic acid, and 2 mg/L BA. The in vitro shoots, 4-5 cm long, were rooted by quick dipping the shoot bases for 3-5 s in 2,000 mg/L indole-3-butyric acid solution followed by transfer to the pots containing farmyard manure, soil, and sand (1:1:1 by volume). Initially, in vitro plantlets were covered with glass jars to maintain a high relative humidity (85-90%). As soon as new shoot growth begins, relative humidity is decreased by exposing them to the open environmental conditions prior transferring to the glasshouse. Indirect shoot regeneration increased the frequency of somaclonal variations. The selected somaclones were used in developing new and novel cultivars.

  15. Micropropagation and hairy root culture of Ophiorrhiza alata Craib for camptothecin production.

    Ya-ut, Pornwilai; Chareonsap, Piyarat; Sukrong, Suchada

    2011-12-01

    An efficient system was developed for the in vitro micropropagation and hairy root culture of Ophiorrhiza alata Craib for camptothecin (CPT) production. Shoot multiplication on leaf and node explants from germinated seeds of O. alata was successful on half-strength Murashige and Skoog medium supplemented with varying amounts of kinetin and α-naphthaleneacetic acid. Node explants grown in vitro were successfully infected by Agrobacterium rhizogenes TISTR 1450 for the establishment of hairy root culture. The amount of CPT in various parts of O. alata was analyzed by HPLC. The accumulation of CPT in transformed hairy roots was twice that in soil-grown plants (785 ± 52 and 388 ± 32 μg/g dry wt, respectively). In the presence of a polystyrene resin (Diaion HP-20) that absorbed CPT, the CPT content in the culture media increased sevenfold compared with controls (1,036 and 151 μg per 250 ml medium, respectively). These results enable the feasible production of CPT of O. alata by means of a cell culture strategy. These measures can help safeguard the plant from extinction. © Springer Science+Business Media B.V. 2011

  16. Preliminary results on in vitro mutagenesis studies on Lansium domesticum Corr

    Mohd Nazir Basiran; Shuhaimi Shamsuddin; Sakinah Ariffin

    2000-01-01

    Dokong (Lansium domesticum Corr.) which belongs to the family Meliaceae is an important fruit trees for the Malaysian fruit industry. Despite the various types that have been identified, the genetic variability is still too narrow for meaningful breeding efforts. Dokong also has long juvenility period and the fruits are pathenocarpically developed. The fruits are often not uniform in size, the tree is prone to bark borers, and the tree architecture needs a lot of pruning for better fruit formation and facilitate easier harvesting. A lot of breeding efforts is needed to improve the genetic characteristics of this species before it can really have an industrial impact. Induced mutation and in vitro culture are two approaches which may be more efficient for genetic improvement. Results of radiosensitivity studies showed that irradiation doses between 50 and 70 Gy is effective enough to induced mutations in seeds. Initial attempts to develop in vitro cultures of Lansium showed that shoot-tips and axillary buds can be cultured to produce plantlets in a medium containing 3 mg/L kinetin and 4 mg/L indoleacetic acid. The procedures can be optimised to develop an efficient micropropagation system. However, attempts to initiate callus cultures have not been successful

  17. Micropropagation of Clerodendrum phlomidis L.F.

    Kher Mafatlal M.

    2016-06-01

    Full Text Available Clerodendrum phlomidis L. f. is an important medicinal plant of the Lamiaceae family, particularly its roots, which are used for various therapeutic purposes in a pulverized form. The objective of this study was to develop a standard protocol for axillary shoot proliferation and rooting of C. phlomidis for its propagation and conservation. Nodal explants were inoculated on Murashige and Skoog (MS medium that was supplemented with one of six cytokinins: 6-benzyladenine, kinetin, thidiazuron, N6-(2-isopentenyl adenine (2iP, trans-zeatin (Zea and meta-topolin. Callus induction, which was prolific at all concentrations, formed at the base of nodal explants and hindered shoot multiplication and elongation. To avoid or reduce callus formation with the objective of increasing shoot formation, the same six cytokinins were combined with 4 μM 2,3,5-tri-iodobenzoic acid (TIBA alone or in combination with 270 μM adenine sulphate (AdS. Nodal explants that were cultured on the medium supplemented with 9.12 μM Zea, 4 μM TIBA and 270 μM AdS produced significantly more and longer shoots than on medium without TIBA and AdS. Half-strength MS medium supplemented with 8.05 μM α-naphthaleneacetic acid was the best medium for root formation. Most (75% in vitro rooted plantlets were successfully acclimatized under natural conditions.

  18. In vitro micropropagation of solanum villosum-a potential alternative food plant

    Iftikhar, A.; Qureshi, R.; Munir, M.

    2015-01-01

    Solanum villosum Miller is annual to biennial herb which is used as potherb as well as fodder/forage that limits its distribution in Pakistan. The aim of this study was to develop a suitable protocol for S. villosum through direct organogenesis. Leaf, stem node and shoot tip explants from the tested plant were inoculated in three different hormonal combinations of BAP (6-benzyl amino purine) alone along NAA (alpha-naphthalene acetic acid) and Kin (Kinetin). Maximum shoot induction was recorded for stem node and leaf (91% each) in MS medium comprising of BAP (1.9 mg/l) and NAA (0.1 mg/l), while shoot tip showed somewhat moderate (81%) response. The highest mean number of shoot (9.1 ± 2 was also obtained for the same medium using leaf explants. Plantlets were successfully rooted in auxin free medium and shifted to green house for multiplication after acclimatizing them. This study may contribute in providing quick and disease free propagation of this neutraceutically and economically potential plant. (author)

  19. Stimulation effect of synthetic cytokinins on the uptake and incorporation of nitrogen-15-labelled ammonium nitrate and urea in wheat leaves

    Iglewski, S.M.; Szarvas, T.; Pozsar, B.I.

    1977-01-01

    The turnover of different labelled nitrogen sources in wheat leaves has been investigated using the isotopic tracer technique. The 15 N at.% was determined in free ammonium ion, in the nitrate and the nitrite levels, and also in the non-disintegrated urea. The accumulation and the incorporation of stable nitrogen was measured in the TCA insoluble protein fraction. According to the experimental data the intensity of incorporation of urea nitrogen is relatively higher than that of the different inorganic compounds. The utilization of ammonium ion was 76% compared with the urea, whereas that of the nitrate nitrogen was 60% in the wheat leaves. The incorporation rate of the two nitrogen atoms from ammonium nitrate was 32% lower than that of the urea nitrogen, in the leaf protein of Bezostaia-1 wheat variety. The turnover of urea through the transamination was very rapid, the amination with ammonium ion was slower, and the first phase of the nitrate reduction was relatively more retarded than the nitrite reduction. The endogenous cytokinin-like biological activity and some synthetic cytokinins (kinetin, benzyladenine) have a remarkably stimulating effect on the incorporation of the different 15 N-labelled nitrogen sources into the leaf protein fraction. (author)

  20. Isozyme modifications and plant regeneration through somatic embryogenesis in sweet potato (Ipomoea batatas (L.) Lam.).

    Cavalcante Alves, J M; Sihachakr, D; Allot, M; Tizroutine, S; Mussio, I; Servaes, A; Ducreux, G

    1994-05-01

    The potential of somatic embryogenesis was evaluated for 10 cultivars of sweet potato through extensive embryogenic response and isozyme analysis. Embryogenic callus was induced by incubating lateral buds on Murashige and Skoog medium containing 10 μM 2,4-dichlorophenoxyacetic acid for 6-8 weeks. The frequency of embryogenic response was low, and varied with genotypes, ranging from 0 to 17%. Embryo to plantlet formation could be enhanced by the use of the combination of 2,4-dichlorophenoxyacetic acid with kinetin, both used at 0.01 μM. Embryogenic callus with its potential of plantlet formation has constantly been maintained for over two years. However, after several subcultures, 0.5 to 12% of embryogenic callus reverted irreversibly into friable fast-growing non-embryogenic callus whose ability to regenerate shoots was then definitively lost. The isozymes of esterase, peroxidase, glutamate oxaloacetate transaminase and acid phosphatase investigated in this study were found appropriate to distinguish compact embryogenic from friable non-embryogenic callus in sweet potato. In fact, the callus reversion was associated with a loss of bands or a decline in isozyme activity. On the contrary, very small changes in isozyme activity or no specific changes at all were observed during the differentiation of embryogenic callus into globular embryos.

  1. Establishment of in vitro adventitious root cultures and analysis of andrographolide in Andrographis paniculata.

    Sharma, Shiv Narayan; Jha, Zenu; Sinha, Rakesh Kumar

    2013-08-01

    Andrographolide is the principal bioactive component of the medicinal plant Andrographis paniculata, to which various diverse pharmacological properties are attributed. Traditionally, andrographolide was extracted from the leaves, stems and other parts of the plant. Leaves have the highest andrographolide content (2-3%) in comparison with the other plant parts. Adventitious root culture of leaf explants of A. paniculata was studied using different strength MS medium supplemented by different concentrations of auxins and a combination of NAA + kinetin for growth and andrographolide production. Among the different auxin treatments in adventitious root culture, only NAA was able to induce adventitious roots. Adventitious roots grown in modified strength MS medium showed the highest root growth (26.7 +/- 1.52), as well as the highest amount of andrographolide (133.3 +/- 1.5 mg/g DW) as compared with roots grown in half- and full-strength MS medium. Growth kinetics showed maximum biomass production after five weeks of culture in different strength MS liquid medium. The produced andrographolide content was 3.5 - 5.5 folds higher than that of the natural plant, depending on the medium strength.

  2. Regulation of root development in Arabidopsis thaliana by phytohormone-secreting epiphytic methylobacteria.

    Klikno, Jana; Kutschera, Ulrich

    2017-09-01

    In numerous experimental studies, seedlings of the model dicot Arabidopsis thaliana have been raised on sterile mineral salt agar. However, under natural conditions, no plant has ever grown in an environment without bacteria. Here, we document that germ-free (gnotobiotic) seedlings, raised on mineral salt agar without sucrose, develop very short root hairs. In the presence of a soil extract that contains naturally occurring microbes, root hair elongation is promoted; this effect can be mimicked by the addition of methylobacteria to germ-free seedlings. Using five different bacterial species (Methylobacterium mesophilicum, Methylobacterium extorquens, Methylobacterium oryzae, Methylobacterium podarium, and Methylobacterium radiotolerans), we show that, over 9 days of seedling development in a light-dark cycle, root development (hair elongation, length of the primary root, branching patterns) is regulated by these epiphytic microbes that occur in the rhizosphere of field-grown plants. In a sterile liquid culture test system, auxin (IAA) inhibited root growth with little effect on hair elongation and significantly stimulated hypocotyl enlargement. Cytokinins (trans-zeatin, kinetin) and ethylene (application of the precursor ACC) likewise exerted an inhibitory effect on root growth but, in contrast to IAA, drastically stimulated root hair elongation. Methylobacteria are phytosymbionts that produce/secrete cytokinins. We conclude that, under real-world conditions (soil), the provision of these phytohormones by methylobacteria (and other epiphytic microbes) regulates root development during seedling establishment.

  3. Effects of gamma-rays irradiation in seed of mungbean (vigna radiata (L.) wilczek) composition of media on shoot regeneration of explants from node of cotyledon

    Hutabarat, Dameria; H, Soeranto

    1998-01-01

    Study the effects of gamma-rays irradiation and composition and media on shoot regeneration of explants from node of cotyledon of mungbean. Wallet variety have been conducted. The explants derived of irradiated seeds of 10-20 Gy of gamma rays were planted in the 0.7% agar solution. One day after planting in the agar media the embryo axis of germinate seed were removed and the node of cotyledon were cultured in the regeneration media as examples. The results shown that shoot regeneration was influenced by media composition and the doses of gamma rays irradiation in seed. In the MURASHIGE and SKOOG medium which contain of BAP or 2-iP or Kinetin with 3 ppm concentrate respectively the explants could produced 100% of shoots. However, the highest. number of produced shoot (3 shoots) was showed in the medium which contained of BAP. The medium with I ppm concentrate od BAP could produced 100% shoot regeneration and the maximum number of shoots (4 shoots) per explant was showed in with 5 ppm. concentrate of BAP. The effectivity off BAP for shoot regeneration by enrichment of 12 ppm Ag 2 SO 4 in the media. Irradiation of 10-20 Gy gamma rays on seeds of mungbean walet variety could improved shoot regeneration of explants from node cotyledon. (author)

  4. Improvement of Salt Tolerance in Trigonella foenum-graecum L. var. PEB by Plant Growth Regulators

    Anjali Ratnakar

    2014-05-01

    Full Text Available The crop yield is reduced under saline conditions and this hampers agricultural productivity. The incorporation of plant growth regulators (PGRs during presoaking treatments in many crops has improved seed performance under saline conditions. In order to study the ameliorative effect of plant growth regulators, experiments were conducted to study the variation in organic constituents in the leaves of Trigonella foenum-graecum L. var.PEB, where the seeds were primed with different plant growth regulators and grown under NaCl salinity. After a pre-soaking treatment of six hours in 20 mg L-1 solutions of gibberllic acid (GA3, 6-furfuryladenine (Kinetin and benzyl adenine (BA, the seeds were allowed to germinate and grow for forty-five days under saline conditions. On the analysis of mature leaves, it was observed that chlorophyll a and b, total chlorophyll and protein showed an increase in PGR-treated plants compared to the untreated set. The accumulation of the stress metabolite such as proline and sugars, which increase under saline conditions, showed a significant decrease in the plants pretreated with PGRs.

  5. Determination of suitable microspore stage and callus induction from anthers of kenaf (Hibiscus cannabinus L.).

    Ibrahim, Ahmed Mahmood; Kayat, Fatimah Binti; Hussin, Zeti Ermiena Surya Mat; Susanto, Dwi; Ariffulah, Mohammed

    2014-01-01

    Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6-8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

  6. Effects of fluoride and 6 benzylaminopurine on growth and respiration of corn and cotton roots

    Thompson, C R

    1967-01-01

    Corn and cotton plants exhibit a wide difference in their susceptibility to atmospheric fluoride. Corn shows leaf lesions when 100 ..gamma../gm on a dry weight basis are accumulated but cotton can tolerate 5000 ..gamma../gm without showing leaf necrosis. A comparison of respirational response of potted seedlings of the two species to 10 ..gamma../M/sup 3/ HF caused an increase of about 10%. Addition of 2 x 10/sup 2/M F/sup -/ to solutions for germinating the plants showed that cotton accumulated about twice as much as F/sup -/ in seedling roots. Growth was reduced about one half by 2 x 10/sup -3/M F/sup -/ in both species but respirational rates of root tips from control and fluoride treated tissues were equal. Prolonged treatment of excised root tips with fluoride reduced respiration. Because fluoride causes cellular changes in roots similar to aging and kinetin seems to act to reverse these changes, corn was germinated with 2 x 10/sup -3/M F/sup -/ and increasing levels of 6-benzylaminopurine. Root growth inhibition (63%) was reversed significantly at 0.2 - 0.8..gamma.. ml. Respirational rates of root tips grown in fluoride, fluoride plus 6-benzylaminopurine and controls were equal.

  7. Regeneration and acclimatization of salt-tolerant arachis hypogaea plants through tissue culture

    Ghauri, E.G.

    2006-01-01

    Excised embryos of Arachis hypogaea were cultured on Murashige and Skoog's medium (MS medium) supplemented with different combinations of growth hormones. The highest frequency of callus proliferation (80%) was recorded on MS medium mixed with 1.0 mg/1 of 2,4-D and 0.5 mg/1 of BAP. These cultures were treated with 0.65 mg/l of trans-4-hydroxy-L-proline (HyP) a:1d various concentrations (0.1-0.5%) of NaCl. In all cases the presence of salt reduced the fresh mass of callus. Shoot regeneration in the cultures took place when transferred to MS medium supplemented with 1.0 mg/1 of kinetin (Kin) and 0.5 mg/1 of 6-benzyl aminopurine (BAP). Percentage of shoot regeneration decreased with the increase of NaCl (0.1- 0.5%) in the shoot regeneration medium. Root formation in these cultures took place when the cultures were nurtured on MS medium free of growth hormones. Regeneration, hardening and acclimatization of the salt tolerant plants was conducted. (author)

  8. Effect of auxin on xylem tracheids differentiation in decapitated stems of Pinus silvestris L. and its interaction with some vitamins and growth regulators

    T. J. Wodzicki

    2015-01-01

    Full Text Available The effects of several vitamins and substances known as important agents in regulation of cell metabolism upon secondary xylem differentiation were studied in interaction with auxin (IAA as applied in lanoline to decapitated stems of 5-year-old Pinus silvestris trees in early and late-summer. Tested substances were: gibberellic acid, kinetin, nicotinic acid, thiamine, pyridoxine, calcium panthotenate, choline chloride, riboflavin, inositol, ascorbic acid, vitamin, A (alcohol, vitamin A (ester, saponin. None of the effects of these substances appeared significant enough to indicate the involvement in the seasonal variation of the response of cambium or differentiating tracheids to auxin. However, several effects, especially those of inositol, vitamin A and pyridoxine upon cambial xylem production and further stages of tracheid differentiation were observed. Auxin (IAA affected cambial activity and subsequent differentiation of tracheids during the earliest stages of cell ontogenesis. At these stages auxin treatment induced quantitative expression of the developmental processes involving radial growth and secondary wall formation by tracheids. In this respect, auxin did not affect cells advanced in differentiation, however, it proved to be an essential factor in the completion of the full cycle of tracheid ontogenesis.

  9. Role of gibberellins and cytokinins in regulation of germination during development and ripening of Triticale caryopses

    Stanisław Weidner

    2014-01-01

    Full Text Available The germination of caryopses of M-T3 Triticale generation, which were freshly harvested in different growth and developmental phases has been studied. A significant influence of the abscisic acid (ABA accumulation on the increment of number of germinating caryopses has been found. Already in the first phase af the embryogenesis considerable stimulating effects of kinetin and gibberellin-A3 (GA3 on the germination of embryos which were isolated from freshly collected grains have been shown. When both stimulators were used together marked synergetic reaction occurred. It has been also determined that in the initial period of embryogensis premature germination occurs, to a higher extent, under the action of cytokinins than! Whether in the further phases of the caryopse development, when embryo develop mainly through the cell elongation, mostly gibberellins seem to be responsible for the activation of germination processes. The more mature were seeds the quicker germinated whole caryopses and embryos isolated from them at different ripeness, after 3-month storage. The highest stimulation of germination by phytohormones has been found for the most mature caryopses. The action of gibberellic acid has been particulary strong.

  10. Plant growth regulators induced urease activity in Cucurbita pepo L. cotyledons.

    El Shora, Hamed M; Ali, Awatif S

    2016-03-01

    This study is aimed to investigate the activity of urease (EC 3.5.1.5, urea amidohydrolase) that catalyzes the hydrolysis of urea in 5-day-old Cucurbita pepo cotyledons subjected to various concentrations of different growth regulators. The treatment of C. pepo cotyledons with different concentrations (100-600 μmol) of different auxins [indole-3-acetic acid (IAA), indole butyric acid (IBA), indole propionic acid (IPA) and naphthalene acetic acid (NAA)]; or with different concentrations (100-300 μmol) of different cytokinins [kinetin, zeatin and benzyladenine (6-BA)] resulted in a significant increase of urease activity, compared to control. The optimal effects were recorded for each of 500 μmol of IAA and 300 μmol of zeatin treatments. A gradual increase in urease activity was detected in cotyledons treated with various concentrations (0.2-1.0 mM) of 28-homobrassinolide (HBL), in relative to control. A substantial increase in urease activity was observed in cotyledons subjected to different concentrations of triazole (10-60 mg L(-1)), containing either triadimefon (TDM) or hexaconazole (HEX), compared to control. The combination of 300 μmol zeatin with any of protein inhibitors, namely 5-fluorouridine (FUrd), cordycepin and α-amanitin, resulted in the alleviation of their inhibitory effect on the urease activity.

  11. Study of some factors affecting potato microtuber production in vitro

    Al-Safadi, B.; Ayyoubi, Z.; Jawdat, D.

    2000-01-01

    The effects of variety, growth regulators, sucrose, and low doses of gamma irradiation on the production of potato (Solanum tuberosum L.) microtubers in vitro were investigated. Nodal segments from virus free explants of 3 potato varieties were placed on six different media and irradiated with 4 doses of gamma radiation (2.5, 5, 10, 15 Gy).Potato varieties used in the study differed in their ability to produce microtubers. Variety Diamant was the best in microtuber production followed by Draga and Spunta. Kinetin played a significant role in inducing tuberization in vitro especially at a concentration of 4 mg -1 . Tuberization was also enhanced by sucrose especially when its level was increased from 3% to 6%. Irradiating the explants with 2.5 Gy of gamma radiation leads to a significant increase in number of microtubers (34% increase over the control). Weight of microtubers was not significantly influenced by low doses of gamma irradiation or media components. Draga microtubers were the largest followed by Diamant and Spunta. Microtubers resembled mature tubers in shape (Spunta was elongated and Draga and Diamant were round). Size of microtubers was crucial for sprouting in vivo. It is suggested that only microtubers larger than 250 mg (5 mm in diameter) can be used to produce minitubers in vivo. Since 2.5 Gy is a low dose, it can be used to enhance tuberization in vitro without fear of genetic changes in the used varieties. (author)

  12. The potential signalling pathways which regulate surface changes induced by phytohormones in the potato cyst nematode (Globodera rostochiensis).

    Akhkha, A; Curtis, R; Kennedy, M; Kusel, J

    2004-05-01

    It has been demonstrated that the surface lipophilicity of the plant-parasitic nematode Globodera rostochiensis decreases when infective larvae are exposed to the phytohormones indole-3-acetic acid (auxin) or kinetin (cytokinin). In the present study, it was shown that inhibition of phospholipase C (PLC) or phosphatidylinositol 3 kinase (PI3-kinase) reversed the effect of phytohormones on surface lipophilicity. The signalling pathway(s) involved in surface modification were investigated using 'caged' signalling molecules and stimulators or inhibitors of different signalling enzymes. Photolysis of the 'caged' signalling molecules, NPE-caged Ins 1,4,5-P3, NITR-5/AM or caged-cAMP to liberate IP3, Ca2+ or cAMP respectively, decreased the surface lipophilicity. Activation of adenylate cyclase also decreased the surface lipophilicity. In contrast, inhibition of PI3-kinase using Wortmannin, LY-294002 or Quercetin, and inhibition of PLC using U-73122 all increased the surface lipophilicity. Two possible signalling pathways involved in phytohormone-induced surface modification are proposed.

  13. GERMINATION AND MULTIPLICATION OF CARNATION USING BAP AND KIN IN VITRO

    D. P. Restelatto

    2014-09-01

    Full Text Available Aiming to establish a system of in vitro multiplication for the harpsichord, the seeds were sterilized in sodium hypochlorite and inoculated in MS. The induction of axillary buds used the BAP and Kinetin (KIN at concentrations of 0; 2,0 and 4,0 mg/L. Was added, 3% (w/v sucrose, 0,6 (w/v agar and the pH ± 5.8. 40 days after the inoculation of seeds, germination was evaluated. The best average germination occurred in T1 (0 min in hypochlorite and T3 (10 min in hypochlorite (9,0 and 8,75 treatments, respectively, differing T2 (5 min in hypochlorite (3,75. In the formation of axillary buds, treatment T1 (2,0 mg / L BAP presented (7,1 buds /explant. Have the treatment T3 (4,0 mg /L BAP presented (5,5 buds / explant, and T1 (zero BAP statistically had the lowest bud formation (3,73 buds / explant. For CIN, it was observed that the best response (5,6 and 5,1 buds/explants were in the T1 (0 CIN and T3 (4.0 mg /L KIN treatments whereas in T2 (2,0 mg /L CIN had the lowest bud formation (2,7 buds /explant . Thus, sodium hypochlorite was effective in decontaminating and BAP and KIN hormones were effective for inducing gems

  14. In vitro propagation of chungah (caralluma tuberculata n.e. brown)

    Rehman, R.

    2014-01-01

    In vitro propagation of Caralluma tuberculata (Chungah) was developed from shoot tip and meristem explants. C. tuberculata is an imperative medicinal plant comprising antidiabetic and anticancer properties. The explants were inoculated on Murashige and Skoog (MS) medium containing different plant growth regulators. Presence of BA or Kin alone in the MS medium did not favor regeneration of shoot from both explants. However, addition of 2,4-dichlorophenoxy acetic acid (2,4-D), gibberellic acid (GA3) and thidiazuron (TDZ) along with 6-benzyl amino purine (BA) or kinetin (Kin) in the medium exhibited significant percentage response, number of shoots per explant and shoot length. Maximum shooting response (53.3+-5.77% from meristem and shoot tip explants each) with highest number of shoots per explant (5.33+-2.08 and 5.6+-2.52 from meristem and shoot tip explants, respectively) were observed at 13.32 micro mol BA along with 2.26 micro mol 2,4-D, 2.89 micro mol GA3 and 9.08*10-3 micro mol TDZ. Replacing BA with kin showed less shoot regeneration response and number of shoots per explant, however, shoots length markedly increased in the presence of Kin. The regenerated plants were successfully rooted and acclimatized in ex vitro condition. The protocol described here can be used for fast multiplication of this endangered herb and genetic transformation. (author)

  15. Effect of Basal Medium, Explants Size and Density on the In Vitro Proliferation and Growth of Date Palm (Phoenix dactylifera L. Cultivar ‘16-bis’

    Mouaad Amine MAZRI

    2013-08-01

    Full Text Available The effect of basal medium, explant size and density on shoot multiplication, growth, rooting and acclimatization of date palm cv. ‘16-bis’ was evaluated. Bud clusters of different sizes (2, 3, 4 and 5 buds per cluster were cultured at density of 1, 2, 3 and 4 clusters on Murashige and Skoog medium (MS, woody plant medium (WPM and Nitsch medium (NM supplemented with 0.5 mg/L 2-naphthoxyacetic acid and 0.5 mg/L kinetin for three months (multiplication phase. Separated shoots of different sizes (<3 cm; 3 to 4.5 cm and 4.5 to 6 cm were cultured at density of 1, 2, 3 and 4 shoots on hormone free MS medium, WPM or NM for three months (Elongation-rooting phase. The proliferation and development of shoots were affected by the basal medium, explant size and density. The optimal shoot proliferation (18.1 was observed when 4 buds clusters were cultured at the density of 2 clusters per jar in MS medium. Separated shoots of 4.5 to 6 cm length exhibited the optimal in vitro development in terms of leaf length and greening, and root number and length when cultured on MS medium. In addition, these shoots reached the highest acclimation frequency with 80%. Our results would be utilized for an efficient propagation of plantlets of cv. ‘16-bis’, a selected date palm cultivar resistant to the bayoud disease.

  16. Establishment of an efficient in vitro regeneration protocol for rapid and mass propagation of Dendrobium chrysotoxum Lindl. using seed culture.

    Nongdam, Potshangbam; Tikendra, Leimapokpam

    2014-01-01

    An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species.

  17. Effect of in vitro mutagenesis on plant regeneration in Citrus aurantifolia S

    Srivastava, R.K.; Sandhu, A.S.; Gosal, S.S.

    2001-01-01

    Callus was induced from different explants excised from in vitro raised seedlings on MS medium enriched with naphthalene acetic acid (NAA) (10 mg/l) and kinetin (0.2 mg/l). The cultures were maintained on the same media for 30 days. Part of the 30-day-old calli were exposed to gamma radiation (5 and 10 Gy) and the rest were treated with ethyl methanesulphonate (EMS) (0.1 to 0.4%) for 8 hours. All the treated calli were immediately transferred to regeneration medium [1/2 MS+Benzyl Amino Purine (BAP) (5 mg/l)] along with the untreated control. The cultures were maintained under conditions of 25±2 deg. C, 16/8 hours day and night regime and 2500-3000 lux light intensity. The results indicated a significant effect of mutagenic agent on callus regeneration and regenerants' morphological features. The same phenomenon was observed in Triticum aestivum and Zea mays. Regenerated mutants showed variation in morphological traits like, plant height, leaf length and breadth. Moreover, the mutants are being screened for resistance against citrus canker. However, the genetic origin of the mutants has not been determined

  18. Phytohormone supplementation significantly increases growth of Chlamydomonas reinhardtii cultivated for biodiesel production.

    Park, Won-Kun; Yoo, Gursong; Moon, Myounghoon; Kim, Chul Woong; Choi, Yoon-E; Yang, Ji-Won

    2013-11-01

    Cultivation is the most expensive step in the production of biodiesel from microalgae, and substantial research has been devoted to developing more cost-effective cultivation methods. Plant hormones (phytohormones) are chemical messengers that regulate various aspects of growth and development and are typically active at very low concentrations. In this study, we investigated the effect of different phytohormones on microalgal growth and biodiesel production in Chlamydomonas reinhardtii and their potential to lower the overall cost of commercial biofuel production. The results indicated that all five of the tested phytohormones (indole-3-acetic acid, gibberellic acid, kinetin, 1-triacontanol, and abscisic acid) promoted microalgal growth. In particular, hormone treatment increased biomass production by 54 to 69 % relative to the control growth medium (Tris-acetate-phosphate, TAP). Phytohormone treatments also affected microalgal cell morphology but had no effect on the yields of fatty acid methyl esters (FAMEs) as a percent of biomass. We also tested the effect of these phytohormones on microalgal growth in nitrogen-limited media by supplementation in the early stationary phase. Maximum cell densities after addition of phytohormones were higher than in TAP medium, even when the nitrogen source was reduced to 40 % of that in TAP medium. Taken together, our results indicate that phytohormones significantly increased microalgal growth, particularly in nitrogen-limited media, and have potential for use in the development of efficient microalgal cultivation for biofuel production.

  19. Study on the presence and influence of phenolic compounds in callogenesis and somatic embryo development of cocoa (Theobroma cacao L..

    Sulistyani Pancaningtyas

    2015-04-01

    Full Text Available Cocoa (Theobroma cacao L. like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D with kinetin (kin. Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.

  20. Isolation and culture of Celosia cristata L cell suspension protoplasts

    Retno Mastuti

    2003-06-01

    Full Text Available Developmental competence of Celosia cristata L. cell suspension-derived protoplasts was investigated. The protoplasts were isolatedfrom 3- to 9-d old cultures in enzyme solution containing 2% (w/v Cellulase YC and 0.5% (w/v Macerozyme R-10 which was dissolvedin washing solution (0.4 M mannitol and 10 mM CaCl2 at pH 5.6 for 3 hours. The highest number of viable protoplasts was releasedfrom 5-d old culture of a homogenous cell suspension. Subsequently, three kinds of protoplast culture media were simultaneously examinedwith four kinds of concentration of gelling agent. Culturing the protoplasts on KM8p medium solidified with 1.2% agarose significantlyenhanced plating efficiency as well as microcolony formation. Afterwards, the microcalli actively proliferated into friable watery calluswhen they were subcultured on MS medium supplemented with 0.3 mg/l 2,4-D and 1.0 mg/l kinetin. Although the plant regenerationfrom the protoplasts-derived calli has not yet been obtained, the reproducible developmental step from protoplasts to callus in thisstudy may facilitate the establishment of somatic hybridization using C. cristata as one parent.

  1. Stable and High Ajmalicine or Serpentine Production of Gamma Radiation Induction Mutant Catharantus Roseus

    Sumaryati Syukur

    2004-01-01

    Catharantus roseus Mutant have been selected by gamma irradiation with 20 krad doses of radiation and characterized as biochemical mutant with anti-feed back inhibition mechanism of tritophan decarboxylase (TDR) enzyme in biosynthetic path way of indole alkaloid. Production of indole alkaloid mainly ajmalicine with high economical values as a pharmaceutical drug for heart attack have been studied by using cell suspension cultures with several variation of medium, elicitors and stress osmosis. This treatment produced variation of indole alkaloid ajmalicine and serpentine. Several induction methods using Murashige and Skoog (MS) medium and polyethylene glycol PEG (6000) 1 to 7%, with hormones concentration of 2,4-D and kinetin as (10 : 1), showed optimal results of ajmalicine range between 20 and 50 nmol/gFW, and serpentine 10 to 60 nmol/gFW. This production increases ten time in mutant (20 Krad) by stress osmotic condition and performed long term stability in culture without subculture. In this paper explanation in detail about the selection methods, stability of mutant and the production of indole alkaloid ajmalicine and serpentine during growth phase, such as adaptation, log, and stationar in suspention culture of mutan cells. (author)

  2. Production of isoflavones, daidzein and genistein in callus cultures of Pueraria candollei Wall. ex Benth. var. mirifica

    Sanha Panichajakul

    2006-03-01

    Full Text Available Callus cultures of Pueraria candollei var. mirifica were first established from various parts of explants with the objective of isoflavones, daidzein and genistein production. The cultures were studied on their growth and isoflavone production by various combinations of growth regulators, auxins and cytokinins at 25±2ºC. Daidzein and genistein accumulated in the cells were determined. The results revealed that callus of P. candollei var. mirifica was capable of producing high level of both isoflavones consistently. The culture temperature played an important role in the growth and isoflavone production. Over twofold of growth and threefold of isoflavone production were demonstrated at 32±2ºC. The callus established from the stems in MS medium supplemented with 4.5 µM 2,4-D and 0.46 µM kinetin produced the highest yield of daidzein (5.12 mg/g, DW and genistein (2.77 mg/g, DW, which was remarkably higher than the intact plants.

  3. Response of different genotypes of wheat, rice and black beans to anther, embryo and other tissue cultures

    Franco, E.; Amador, D.; Calderon, J.; Alvarez, G.; Alvarado, J.; Ramazzini, H.; Ramos, S.; Acuna, G.; Zuniga, B.

    1996-01-01

    The objective of the basic studies we have been conducting in our laboratory is to establish callus induction and in vitro plant regeneration protocols starting with several tissues of Guatemalan varieties of wheat (Triticum aesticum L.), rice (Oryza sativa L.) and especially black bean (Phaseolus vulgaris L.) in order to obtain disease resistance, earliness, and dwarf plants. Wheat anthers and immature embryos of varieties Patzun, Comalapa, Chocoyo, and Xequijel cultured in N 6 , Potato II, and MS basal media supplemented with auxin and cytokinin gave the best responses in callus induction and plant regeneration. Anthers and mature embryos of indica rice varieties Precozicta and Virginai, when cultured in MS, B 5 , N 6 , and Potato II basal media with different hormonal combinations gave a good response in callus induction. However, a satisfactory response in plant regeneration was not obtained. With black beans, when hypocotyls and mature embryos of black bean varieties Quinack Che and Parramos were cultured in MS basal medium supplemented with different concentrations of NAA and kinetin, more than 60% callus induction was produced. When Quinack Che calli were transferred to MS basal medium supplemented with 1 mg/l NAA plus 0.5 mg/l BAP, green points of regeneration were visible in these calli. (author). 34 refs, 28 tabs

  4. Tissue culture and mutagenesis of rain lily (zephyranthes)

    Mohd Nazir Basiran; Zaiton Ahmad; Shakinah Salleh; Shuhaimi Shamsudin; Aiza Shaliha Jamaludin

    2004-01-01

    There are three varieties of Zephyranthes used widely in landscaping due to their robust growth and attractive flowers in pink, yellow and white. Both in vivo and in vitro mutagenesis are an effective approach to increase the flower colour variations of Zephyranthes. In vitro propagation for the three varieties was attempted by using the induction medium developed by Sachar and Kapoor in 1959. The medium contains I ma of each indole 3-acetic acid (IAA), indole 3-butyric acid (IBA) and kinetin. Following surface sterilization of bulb scales, 17.8%, 10.5% and 10.7% of pink, white and yellow varieties respectively, were able to form small bulblets on the induction media. Further development of these bulblets into plantlets was also achieved on the same medium. Work is now being carried out to improve the efficiency of bulblet regeneration. Mutagenesis of Zephyranthes was initiated from bulbs of the pink varieties to develop new varieties with attractive combinations of flower colour and forms, shelf life and growth habits. These bulbs were irradiated using a gamma cell with a 60 Co source. Three variants with different flower colour and morphology have been achieved so far and are now being propagated in the nursery. (Author)

  5. Assessment of genetic variability in somaclonal population of sugarcane

    Seema, N.; Raza, S.; Yasmeen, S.; Bibi, S.; Nizamani, G.S.

    2014-01-01

    In the present study plant tissue culture technique was used to create the genetic variability in three sugarcane clones NIA98, BL4 and AEC82-1026. Callus induced in these clones in media containing MS + 2, 4 D (2mg 1it-1) and Dicamba (1mg1it-1). The embryogenic calli then regenerated in media containing MS basal media + Kinetin (2mg1it-1) + IBA (2mg1it-1) + IAA (2mglit-1). After shooting and rooting, plants were exposed to green house and acclimatization of the somaclones in the field condition. RAPD markers were used to evaluate the genetic variation at DNA level between parents and somaclones of NIA98, BL4 and AEC82-1026 developed through callus culture. Fourteen RAPD primer chosen randomly were used to amplify DNA from plant material to assess the genetic variation between parents and regenerated somaclones. The highest similarity was obtained between BL4 parent and BL4 somaclone (96%). While minimum similarity found between NIA-98 parent and AEC82-1026 somaclone (69%). In this study, we used RAPD to investigate the somaclonal variation in sugarcane clones derived from callus cultures. (author)

  6. Increased production of azadirachtin from an improved method of androgenic cultures of a medicinal tree Azadirachta indica A. Juss.

    Srivastava, Priyanka; Chaturvedi, Rakhi

    2011-07-01

    Present report is the first direct evidence of azadirachtin production in androgenic haploid cultures of Azadirachta indica, a woody medicinal tree. Anther cultures at early-late-uninucleate stage of microspores were established on MS medium with BAP (5 μM), 2,4-D (1 μM) and NAA (1 μM) containing 12% sucrose. The calli, induced, were further multiplied on 2,4-D and Kinetin media. Shoots, differentiated on BAP (2.2 μM) + NAA (0.05 μM) medium, were elongated on MS + BAP (0.5 μM) and multiplied on MS + BAP (1 μM) + CH (250 mg/l). Thereafter, the shoots were rooted on ¼ MS + IBA (0.5 μM). Cytological analysis of the calli and regenerants have confirmed their haploid status with the chromosome number as 2n = x = 12. The haploid cell lines and leaves from in vitro grown plantlets were analyzed for azadirachtin by RP-HPLC and mass spectroscopy. Maximum azadirachtin (728.41 μg/g DW) was detected in calli supporting best shoot proliferation while least (49 μg/g DW) was observed in an undifferentiated line from maintenance medium. This study has brought us a step closer to develop genetically pure lines that could serve as new and attractive alternative ways of homogeneous controlled production of high value compounds, round the year, independent of geographical and climatic barrier.

  7. Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L. (Scrophulariaceae)-an ethnomedicinal herb.

    Premkumar, G; Sankaranarayanan, R; Jeeva, S; Rajarathinam, K

    2011-06-01

    To develop an improved protocol for micropropagation of ethnomedicinally important Scoparia dulcis (S. dulcis) L. Explants were inoculated on MS basal medium supplemented with kinetin and 6-benzylaminopurine for shoot bud induction. To enhance the shoot induction, various auxins like 3-indoleacetic acid or 3-indolebutyric acid or α-naphthylacetic acid were tested along with 2.32 M KI and 4.44 µM BAP. The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IAA, IBA or NAA. After roots were developed, the plantlets were transplanted to pots filled with vermiculate and sand and kept in growth chamber with 70%-80% humidity under 16 h photoperiod. After acclimatization, the plantlets were transferred to the garden and survival percentage was calculated. Data were statistically analyzed and means were compared using Duncan's multiple range test (Pdulcis. Shoot induction on young leaf explants was most successful in MS medium supplemented with combination of two cytokinins (2.32 µM KI and 4.44 µM BAP) 2.85 µM IAA, 10% CM and 1 483.79 µM adenine sulfate. A single young leaf explant was capable of producing 59 shoots after 13 days of culture. Flower was induced in medium supplemented with combination of KI and BAP. Cytokinins are the key factor to induce the direct shoot regeneration and flowering of S. dulcis.

  8. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

    Sutini Sutini

    2017-02-01

    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  9. ADAPTATION OF THE OBTAINED in vitro Gentiana lutea L. PLANTS TO ex vitro AND in situ CONDITIONS

    О. Yu.

    2015-12-01

    Full Text Available The objective of the research was to develop the technology of introduction of the obtained by microclonal propagation Gentiana lutea L. plants into conditions in situ. Methods of cultivation of plant objects in vitro were used. There were chosen optimal conditions for rooting G. lutea shoots obtained through microclonal propagation in vitro: МS/2 medium with twice decreased concentration of NH4NO3 without vitamins and sucrose supplemented with 3 g/l of mannite and 0.05 mg/l kinetin, and agar (4 mg/l in combination with perlite (16 g/l used as a maintaining substrate; or the nutrient medium (MS/2 without vitamins and smaller concentration of N4NO3 with gradual decrease of carbohydrates from 10 g/l to 2 g/l, and further rooting experimental shoots in tap water. Rooted plants were adapted to conditions ex vitro through planting them into flowerpots with soil and gradual changing hothouse regime for exposed one. The share of adapted to in situ conditions plants (21% after a year of planting proves the suggested method to be efficient and promising. There was suggested this technology is the most efficient ones for revival of disturbed G. lutea populations that includes repatriation of rooted and adapted to ex vitro conditions plants obtained through microclonal propagation in vitro.

  10. Direct organogenesis and leaf-anatomy modifications in vitro of neoregelia concentrica (vellozo) l.b. smith (bromeliaceae)

    Martins, J.P.R.; Castro, E.M.D.; Jani, T.F.; Pasqual, M.; Schmildt, E.R.

    2014-01-01

    Tissue culture can contribute in the multiplication of several species with commercial interest, like the bromeliads. It was aimed to evaluate cytokinins and its concentrations in the multiplication and leaf structure of Neoregelia concentrica (Vellozo) L.B. Smith. Previously In vitro-established N. concentrica plants were inoculated in MS medium supplemented with 6-benzylaminopurine (BAP) or Kinetin (KIN) with concentrations 0.0, 5.0, 10.0 and 15.0 micro M. For the anatomic analyses tree plants of each treatment were randomly sampled at 60-day growth. Significant differences were verified in the evaluated characteristics due to the treatments. The raise in cytokinin concentrations induced a higher percentage and average number of explants with shoots. BAP provided higher averages when compared to KIN. The cytokinin use modified the epidermal structure and induced a larger thickening of the water-storage and chlorophyll parenchymas. The use of 15.0 micro M BAP was efficient in the In vitro multiplication and in the leaf tissue development of N. concentrica. (author)

  11. Effects of Plant Growth Hormones on Mucor indicus Growth and Chitosan and Ethanol Production.

    Safaei, Zahra; Karimi, Keikhosro; Golkar, Poorandokht; Zamani, Akram

    2015-07-22

    The objective of this study was to investigate the effects of indole-3-acetic acid (IAA) and kinetin (KIN) on Mucor indicus growth, cell wall composition, and ethanol production. A semi-synthetic medium, supplemented with 0-5 mg/L hormones, was used for the cultivations (at 32 °C for 48 h). By addition of 1 mg/L of each hormone, the biomass and ethanol yields were increased and decreased, respectively. At higher levels, however, an inverse trend was observed. The glucosamine fraction of the cell wall, as a representative for chitosan, followed similar but sharper changes, compared to the biomass. The highest level was 221% higher than that obtained without hormones. The sum of glucosamine and N-acetyl glucosamine (chitin and chitosan) was noticeably enhanced in the presence of the hormones. Increase of chitosan was accompanied by a decrease in the phosphate content, with the lowest phosphate (0.01 g/g cell wall) being obtained when the chitosan was at the maximum (0.45 g/g cell wall). In conclusion, IAA and KIN significantly enhanced the M. indicus growth and chitosan production, while at the same time decreasing the ethanol yield to some extent. This study shows that plant growth hormones have a high potential for the improvement of fungal chitosan production by M. indicus.

  12. Thidiazuron enhanced regeneration and silymarin content in silybum marianum

    Khan, M.A.; Abbasi, B.H.; Shinwari, Z.K.

    2014-01-01

    Silybum marianum, of family Asteraceae is renowned for production of biologically important silymarin, which has shown multi-dimensional medicinal properties. It has a high protective role against jaundice and hepatitis C worldwide. We hereby established a feasible and efficient method for indirect regeneration of S. marianum for production of consistent plantlets. Calli were induced from leaf explants of seed-derived plantlets on Murashige and Skoog (MS) medium supplemented with several concentrations of different plant growth regulators (PGRs). Highest callogenic response (89%) was recorded for 4.4 meu M Thidiazuron (TDZ) in combination with 6.6 meu M Kinetin (Kn). Subsequent sub-culturing of callus after 4 weeks of culture, on medium with similar compositions of PGRs induced shoot organogenesis. Highest shoot induction frequency (86%) with maximum mean multiple shoots (26 shoots per explant) were recorded for 11meu M TDZ after 4 weeks of transfer. Longest shoots (4.1 cm) were recorded for MS medium augmented with 6.6 meu M TDZ and 4.4 meu M naphthalene acetic acid (NAA). Furthermore, rooted plantlets were developed on MS medium containing different concentrations of indole acetic acid (IAA). Silymarin was determined by High performance liquid chromatography (HPLC) and 8.47 mg/g DW silymarin was detected in the regenerated plantlets. This study contributes to a better understanding of the different mechanisms involved in morphogenesis and production of biologically active principle in Silybum marianum. (author)

  13. Top 10 botanical ingredients in 2010 anti-aging creams.

    Cronin, Hyland; Draelos, Zoe Diana

    2010-09-01

    New developments in the realm of skin rejuvenation such as phytotherapy are at an astounding increasing pace in the cosmeceutical market. Yet, many of these products that are classified as cosmeceuticals are tested less vigorously and do not have to be approved by the Food and Drug Administration to establish efficacy and safety. Thus, as clinicians, we must ask the question, "Is there science-based evidence to validate the mechanism of these new treatments?" We assessed the top anti-aging creams currently on the market specifically evaluating their botanical ingredients. Some of the most common botanicals that are hot off the market are: Rosmarinus officinalis, Vitis vinifera (grape seed extract), Citronellol, Limonene, Oenothera biennis (evening primrose), Glycyrrhiza glabra (licorice extract), Aframomum angustifolium seed extract, Diosgenin (wild yam), N6 furfuryladenine (kinetin), and Ergothioneine. Through researching each of these botanical ingredients, we have concluded that randomized controlled trials are still needed in this area, but there is promise in some of these ingredients and science to validate them. © 2010 Wiley Periodicals, Inc.

  14. The Use of Maleic Hydrazide for Effective Hybridization of Setaria viridis.

    Govinda Rizal

    Full Text Available An efficient method for crossing green foxtail (Setaria viridis is currently lacking. S. viridis is considered to be the new model plant for the study of C4 system in monocots and so an effective crossing protocol is urgently needed. S. viridis is a small grass with C4-NADP (ME type of photosynthesis and has the advantage of having small genome of about 515 Mb, small plant stature, short life cycle, multiple tillers, and profuse seed set, and hence is an ideal model species for research. The objectives of this project were to develop efficient methods of emasculation and pollination, and to speed up generation advancement. We assessed the response of S. viridis flowers to hot water treatment (48°C and to different concentrations of gibberellic acid, abscisic acid, maleic hydrazide (MH, and kinetin. We found that 500 μM of MH was effective in the emasculation of S. viridis, whilst still retaining the receptivity of the stigma to pollination. We also report effective ways to accelerate the breeding cycle of S. viridis for research through the germination of mature as well as immature seeds in optimized culture media. We believe these findings will be of great interest to researchers using Setaria.

  15. Bioengineering of photosynthetic membranes. Requirement of magnesium for the conversion of chlorophyllide a to chlorophyll a during the greening of etiochloroplasts in vitro

    Daniell, H.; Rebeiz, C.A.

    1984-01-01

    The massive conversion of delta-aminolevulinic acid (ALA) to protochlorophyllide (Pchlide) and the massive conversion of chlorophyllide a (Chlide a) to chlorophyll a (Chl a) are two essential conditions for the ALA-dependent assembly of photosynthetic membranes in vitro. In this work, the authors describe the development of a cell-free system capable of the forementioned biosynthetic activities at rates higher than in vivo, for the first 2 h of dark-incubation. The cell-free system consisted of 1) etiochloroplasts prepared from kinetin and gibberellic-acid-pretreated cucumber cotyledons, and 2) cofactors and additives described elsewhere and which are needed for the massive conversion of ALA to Pchlide, 3) high concentrations of ATP, MgCl/sub 2/, and an isoprenol alcohol such as phytol, were required for the massive conversion of Chlide a to Chl a. An absolute and novel requirement of Mg/sup 2 +/ for the conversion of Chlide a to Chl a was also demonstrated. In addition to the role of phytol as a substrate for the conversion of Chlide a to Chl a, the data suggested that this alcohol may also be involved in the regulation of the reactions between ALA and Pchlide. It is proposed that during greening, the conversion of Chlide a to Chl a may follow different biosynthetic rates, having different substrate and cofactor requirements, depending on the stage of plastid development.

  16. Gamma irradiation effect on the formation of Clonal variation from catharantus roseus plant

    Syukur, Sumaryati

    2000-01-01

    Clonal variation have been found in Catharantus roseus plant after gamma irradiation. Several doses have been used to produce clonal variation. The most effective doses used to perform better clonal variation was 20 krad. About 103 seeds irradiated for every radiation treatment, but only several clones were grown better than wild type. We have success to get (M) seeds the expected mutant. The seeds from selected mutant are bigger when compare to the wild type and growth better on medium containing 5-methyl Tryptophan (5-MT). The chlorophyll content is higher (almost twice) as compared to the wild type. Fulther experiment continue to do in vitro culture in order to develop embryonic callus from leaf tip and leaf base. Several manipulation of auxin and cytokini have been used to differentiate the callus formation. Modified MS medium with kinetin and cytokinin (10:1) can induce globular embryo like structure. Dragendrof alkaloid reagent were used to determine high alkaloid clones from the expected mutant. TLC analysis from callus mutant shows 3 clear bands with subsequence Rf about 0.22, 0.58 while control shows two smearing bands at 0.21 and 0.52

  17. Regeneration of some monocotyledonous plants from subterranean organs in vitro

    Krystyna D. Kromer

    2013-12-01

    Full Text Available The aim in view was investigation ofthe regenerative potential of rhizomes, bulbs and corms of ten monocotyledonous plant species from four families: Amaryllidaceae (Haemanthus katharinae, Crinum abyssinicum, Leucojum vernum, Araceae (Spathiphyllum wallisii, Iridaceae (Crocus vernus, Iris germanica, Liliaceae (Hosta lancifolia, Muscari racemosum, Scilla laxiflora, Veltheimia viridifolia under conditions of in vitro culture. All the investigated species were capable of buld or, alternatively, bud and root regeneration. Different morphogenetic potential was noted between the particular families. A high regenerative potential under the conditions of culture applied was characteristic for plants of the Liliaceae and Araceae families, it was lower in plants belonging to Amaryllidaceae and lowest in those of the Iridaceae family. Plants from the Liliaceae family exhibited also the highest ability of callus formation, whereas Amaryllidaceae and Iridaceae plants possessed this ability in only a low degree. The influence of growth regulators of the auxin group (NAA, IAA, 2,4-D and of cytokinins (K on the initiation and course of organogenesis was tested. The results of the experiments indicate that auxins in interaction with kinetin gave the highest percentage of regenerating explants and also a large number of buds on the latter. Stimulation of callus tissue was highest under the influence of 2,4-D, and weaker when NAA IAA were used.

  18. In vitro propagation of critically endangered species Scilla autumnalis L. – biochemical analyses of the regenerants

    Cristian BANCIU

    2010-11-01

    Full Text Available The present study belongs to the international efforts for plant conservation from the areas threatened by human activities. The saline soils areas are restricting for agriculture and in some cases for fishery facilities and the plant species are extinct from those areas. Scilla autumnalis L. is one of the threatened plants (rare on the national red list of vascular plants from Romania that grows in the Natural Park Comana, Giurgiu County, South Romania. Seeds from plants grown in the natural habitat have been used for in vitro plant regeneration and multiplication. After successfully rooting and acclimatization of the regenerated plantlets from germinated seeds, biochemical studies have been performed in order to compare the regenerants from in vitro cultures with native plants from genetically point of view. Peroxydase and esterase’s spectra were the biochemical markers used.The results indicated that this plant species can be multiplicated, rooted and acclimatized on synthetic medium (MS supplemented with NAA, IBA, IAA, kinetin and BAP with a good efficiency and the regenerants had no genetic alterations determinated by culture conditions.

  19. Effect of Acute and Chronic Gamma Irradiation on in vitro Growth of Stevia rebaudiana Bertoni

    Norazlina Noordin; Rusli Ibrahim; Nur Hidayah Mohd Sajahan; Salmah Moosa; Sobri Hussein

    2014-01-01

    Stevia rebaudiana Bertoni is a perennial herb that belongs to the family of Asteraceae. It is a natural sweetener plant known as sweet leaf, which is estimated to be 300 times sweeter than cane sugar. In this study, micropropagation and in vitro mutagenesis of this natural herb was successfully conducted. It was found that shoot tips on MS medium supplemented with 1 mg/l Kinetin showed the highest shoot induction and multiplication after 3 weeks of culture. Radiosensitivity test was conducted to identify the LD50 for in vitro stevia shoots and to select effective doses to be used for the in vitro mutagenesis. Shoot tips were irradiated with acute and chronic gamma radiation at 0, 10.00, 20.00, 30.00, 40.00, 60.00, and 80.00 Gy. At 60 Gy and 80 Gy, the shoot tips demonstrated 0 % survival, all were killed. LD 50 for stevia (the dose that killed 50 % of the irradiated explants) was at 29 Gy. In this study, LD 50 for the stevia (the dose that killed 50 % of the irradiated explants) was at 29 Gy for acute irradiation and was at 45 Gy for chronic irradiation. The effective doses were selected at 10, 20 and 30 Gy. These three selected doses were applied for the in vitro mutagenesis of the stevia shoots. (author)

  20. Antimicrobial activity of Anonna mucosa (Jacq. grown in vivo and obtained by in vitroculture

    Thiago José de Souza Barboza

    2015-09-01

    Full Text Available Brazilian flora includes numerous species of medicinal importance that can be used to develop new drugs. Plant tissue culture offers strategies for conservation and use of these species allowing continuous production of plants and bioactive substances. Annona mucosa has produced substances such as acetogenins and alkaloids that exhibit antimicrobial activities. The widespread use of antibiotics has led to an increase in multidrug-resistant bacteria, which represents a serious risk of infection. In view of this problem, the aim of this work was to evaluate the antibacterial potential of extracts of A. mucosa obtained by in vitro techniques and also cultured under in vivo conditions. Segments from seedlings were inoculated onto different culture media containing the auxin picloram and the cytokinin kinetin at different concentrations. The calluses obtained were used to produce cell suspension cultures. The materials were subjected to methanol extraction and subsequent fractionation in hexane and dichloromethane. The antimicrobial activity against 20 strains of clinical relevance was evaluated by the macrodilution method at minimum inhibitory and minimum bactericidal concentrations. The extracts showed selective antimicrobial activity, inhibiting the growth of Streptococcus pyogenes and Bacillus thuringiensis at different concentrations. The plant tissue culture methods produced plant materials with antibacterial properties, as well as in vivo grown plants. The antibacterial activity of material obtained through biotechnological procedures of A. mucosa is reported here for the first time.

  1. Evaluation of different carbon sources for high frequency callus culture with reduced phenolic secretion in cotton (Gossypium hirsutum L. cv. SVPR-2

    G. Prem Kumar

    2015-09-01

    Full Text Available An efficient protocol was developed to control excessive phenolic compound secretion during callus culture of cotton. As cotton is naturally rich in phenolic compounds factors influencing the phenolic compound secretion, callus induction and proliferation were optimized for getting high frequency callus culture. Different carbon sources such as fructose, glucose, sucrose and maltose were tested at various concentrations to control phenolic secretion in callus culture. Among them, 3% maltose was found to be the best carbon source for effectively controlling phenolic secretion in callus induction medium. High frequency of callus induction was obtained on MSB5 medium supplemented with 3% Maltose, 2,4-D (0.90 μM and Kinetin (4.60 μM from both cotyledon and hypocotyl explants. The best result of callus induction was obtained with hypocotyl explant (94.90% followed by cotyledon explant (85.20%. MSB5 medium supplemented with 2,4-D (0.45 μM along with 2iP (2.95 μM gave tremendous proliferation of callus with high percentage of response. Varying degrees of colors and textures of callus were observed under different hormone treatments. The present study offers a solution for controlling phenolic secretion in cotton callus culture by adjusting carbon sources without adding any additives and evaluates the manipulation of plant growth regulators for efficient callus culture of SVPR-2 cotton cultivar.

  2. Establishment of an Efficient In Vitro Regeneration Protocol for Rapid and Mass Propagation of Dendrobium chrysotoxum Lindl. Using Seed Culture

    2014-01-01

    An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species. PMID:25401154

  3. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.

    Ahmed Mahmood Ibrahim

    2014-01-01

    Full Text Available Kenaf (Hibiscus cannabinus L. is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

  4. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.)

    Binti Kayat, Fatimah; Ermiena Surya Mat Hussin, Zeti; Susanto, Dwi; Ariffulah, Mohammed

    2014-01-01

    Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA. PMID:24757416

  5. Photochemical oxidants injury in rice plants. III. Effect of ozone on physiological activities in rice plants

    Nakamura, H; Saka, H

    1978-01-01

    Experiments were made to determine the effect of photochemical oxidants on physiological activities of rice plants. Rice plants were fumigated with ozone at concentrations of 0.12-0.20 ppm for 2-3 hr to investigate acute injury and at 0.05 and 0.09 ppm for daily exposure from 3.0 leaf stage to assess the effect of ozone on growth. It was observed that malondialdehyde produced by disruption of the components of the membrane increased in the leaves exposed to ozone. Ozone reduced the RuBP-carboxylase activity in both young and old leaves 12-24 hr after fumigation. In the young leaves the activity of this enzyme recovered to some extent after 48 hr, but it did not show any recovery in the old leaves. On the other hand, ozone remarkably increased the peroxidase activity and slightly increased acid phosphatase in all leaves. Abnormally high ethylene evolution and oxygen uptake were detected in leaves soon after ozone fumigation. In general, high molecular protein and chlorophyll contents in the detached leaves decreased with incubation in dark, particularly in the old ones. These phenomena were more accelerated by ozone fumigation. Kinetin and benzimidazole showed significant effects on chlorophyll retention in ozone-exposed leaves. Reduction of plant growth and photosynthetic rate was recognized even in low concentration of ozone in daily exposure at 0.05 and 0.09 ppm. From these results it was postulated that ozone may cause the senescence of leaves in rice plants.

  6. Accumulation of carbohydrates in the development of tomato plants treated with different chemical products

    Anamaria Ribeiro Pereira Ramos

    2015-04-01

    Full Text Available This work had the purpose to study the physiological effects of pyraclostrobin, boscalid, plant growth regulators and plant extract on the accumulation of carbohydrates during the development of tomato plants (Solanum lycopersicum L., hybrid Giuliana, in protected environment conditions. The treatments were: T1- control; T2- pyraclostrobin 0.2 g L-1; T3- boscalid 0.075 g L-1, T4- pyraclostrobin 0.2 g L-1 + boscalid 0.075 g L-1, T5- IBA + GA3 + kinetin 375 mg L-1, T6- GA4+7 + benzylaminopurine 100 mg L-1 and T7- plant extract 100 mg L-1. The carbohydrate accumulation curve was accomplished with 5 samples, at 20-day intervals between evaluations, the 1st evaluation being carried out at 30 days after transplantation, on the day of the first treatment application. At each sampling the plants were separated in stem, leaves and fruits, of which the contents of total soluble sugars, reducing sugars and saccharose were evaluated. The effects of the treatments on chlorophyll content and gas exchanges were also evaluated. The experimental design was completely randomized, with 4 repetitions and 6 destructive evaluations during the development, with 1 plant per experimental unit for each sampling. The pyraclostrobin and boscalid applied in isolation and/or combined favor the increase of carbohydrates in leaves, stems and fruits of tomato hybrid Giuliana

  7. In vitro regeneration from petiole explants of non-toxic Jatropha curcas

    Kumar, Nitish

    2011-01-01

    Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 mu M TDZ. The Induced shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 mu M BA and 8.5 mu M IAA. The elongated shoots could be rooted on half-strength MS medium with 15 mu M IBA, 11.4 mu M IAA and 5.5 mu M NAA with more than 90% survival rate. (C) 2010 Elsevier B.V. All rights reserved.

  8. Micropropagation of Bioencapsulation and Ultrastructural Features of Sainfoin (Onobrychis viciifolia Grown In Vivo and In Vitro

    Sadegh Mohajer

    2014-01-01

    Full Text Available To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601 of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L. were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0–2.0 mg L−1 and Indole-3-acetic acid (0–2.0 mg L−1 was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1 : 1 had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial, 0.188 (adaxial in in vivo and 0.121 (abaxial, 0.201 (adaxial in in vitro leaves.

  9. In vitro propagation of cauliflower using curd microexplants.

    Kieffer, Martin; Fuller, Michael P

    2013-01-01

    Cauliflower (Brassica oleracea var. botrytis) with its distinctive pre-inflorescence or curd is a remarkable member of the Brassica cabbage group. During curd development, intense and repetitive branching leads to a spectacular increase in size and the accumulation of millions of meristems at its surface. Although destined to produce flowers, most of these meristems are capable of regenerating vegetative shoots in vitro, making curd fragments an excellent material for the micropropagation of cauliflower. Most reported methods using these tissues were devised for the production of small clones of vitroplants as the true potential of curd fragments remained highly underestimated. We describe a technique exploiting fully this abundance of meristems and optimized for the large-scale in vitro propagation of cauliflower. The curd surface is first mechanically disrupted to break up the meristem clusters and generate microexplants carrying 1-3 meristems. These microexplants are then cultured at high density 1:100 (v:v) (microexplants:medium) in liquid medium containing Kinetin and indole-3-butyric acid (IBA) and produce thousands of microshoots in 12 days. After selecting the best quality microshoots on a sucrose pad, they are transferred en masse to a rooting medium supplemented with IBA. Four weeks later, rooted microshoots are carefully acclimatized before transfer to the field. This semi-automated protocol is rapid, cost effective, and well adapted for the production of clones of several thousands of plants by a single worker in a short space of time.

  10. Comparison of Cuminaldehyde Contents from Cell Suspension Cultures and Seeds of [Bunium persicum (Boiss. B. Fedtsch.

    Sara KHOSRAVINIA

    2012-11-01

    Full Text Available The cell suspension culture and seed samples of Bunium persicum were extracted by supercritical fluid, hydrodistillation and solvent methods and analyzed by Gas Chromatography. In this study to compare the different methods of extractions, cuminaldehyde was targeted as one of the Black zira essential oil constitute. For callus induction the germinated seeds were cultured as explants on Murashige and Skoog medium supplemented with 2 mg/l 2,4-dichlorophenoxy acetic acid and 0.5 mg/l kinetin (treatment A as well as 2 mg/l ?-naphthalene acetic acid and 0.5 mg/l 6-benzyl aminopurine (treatment B and followed by cells suspension cultures establishment for the first time. The results of cell culture showed that cells from treatment B have a growth rate higher than A. All extracts were dissolved in 1 ml hexane and analyzed by Gas Chromatography. According to the Gas Chromatography analysis, cuminaldehyde was not detected in the supercritical fluid samples, while it was present in hydrodistillation and solvent extract. Cuminaldehyde percentage in cell and seed solvent extracts was 4.65% and 18.61% respectively. Gas Chromatography results also showed that no cuminaldehyde is present in media extracts, means no cuminaldehyde has been secreted into the medium.

  11. Detection of biosynthetic gene and phytohormone production by endophytic actinobacteria associated with Solanum lycopersicum and their plant-growth-promoting effect.

    Passari, Ajit Kumar; Chandra, Preeti; Zothanpuia; Mishra, Vineet Kumar; Leo, Vincent Vineeth; Gupta, Vijai Kumar; Kumar, Brijesh; Singh, Bhim Pratap

    2016-10-01

    In the present study, fifteen endophytic actinobacterial isolates recovered from Solanum lycopersicum were studied for their antagonistic potential and plant-growth-promoting (PGP) traits. Among them, eight isolates showed significant antagonistic and PGP traits, identified by amplification of the 16S rRNA gene. Isolate number DBT204, identified as Streptomyces sp., showed multiple PGP traits tested in planta and improved a range of growth parameters in seedlings of chili (Capsicum annuum L.) and tomato (S. lycopersicum L.). Further, genes of indole acetic acid (iaaM) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase (acdS) were successively amplified from five strains. Six antibiotics (trimethoprim, fluconazole, chloramphenicol, nalidixic acid, rifampicin and streptomycin) and two phytohormones [indole acetic acid (IAA) and kinetin (KI)] were detected and quantified in Streptomyces sp. strain DBT204 using UPLC-ESI-MS/MS. The study indicates the potential of these PGP strains for production of phytohormones and shows the presence of biosynthetic genes responsible for production of secondary metabolites. It is the first report showing production of phytohormones (IAA and KI) by endophytic actinobacteria having PGP and biosynthetic potential. We propose Streptomyces sp. strain DBT204 for inoculums production and development of biofertilizers for enhancing growth of chili and tomato seedlings. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  12. In vitro propagation, ex vitro rooting and leaf micromorphology of Bauhinia racemosa Lam.: a leguminous tree with medicinal values.

    Sharma, Udit; Kataria, Vinod; Shekhawat, N S

    2017-10-01

    A micropropagation system for Bauhinia racemosa Lam. was developed involving axillary shoot proliferation and ex vitro rooting using nodal explants obtained from mature tree. MS medium with 3.0 mg l -1 BA (6-benzyladenine) was optimum for shoot bud induction. For shoot multiplication, mother explants were transferred repeatedly on medium containing low concentration of BA (0.75 mg l -1 ). Number of shoots was increased up to two passages and decreased thereafter. Shoot multiplication was further enhanced on MS medium containing 0.25 mg l -1 each of BA and Kin (Kinetin) with 0.1 mg l -1 of NAA (α-naphthalene acetic acid). Addition of 0.004 mg l -1 TDZ (thidiazuron) increased the rate of shoot multiplication and 21.81 ± 1.26 shoots per culture vessel were obtained. In vitro regenerated shoots were rooted under ex vitro conditions treated with 400 mg l -1 IBA (indole-3-butyric acid) for 7 min on sterile soilrite. After successful hardening in greenhouse, ex vitro rooted plants were transferred to the field conditions with ≈85% of survival rate. Micromorphological changes were observed on leaf surface i.e. development of vein density and trichomes and stomatal appearance, when plants were subjected to environmental conditions. This is the first report on in vitro regeneration of B. racemosa from mature tree.

  13. An efficient method for in vitro callus induction in Myrciaria dubia (Kunth Mc Vaugh "Camu Camu"

    Ana M. Córdova

    2014-03-01

    Full Text Available Due to the high variability in vitamin C production in Myrciaria dubia "camu camu", biotechnological procedures are necessary for mass clonal propagation of promising genotypes of this species. The aim was to establish an efficient method for in vitro callus induction from explants of M. dubia. Leaf and knot sex plants were obtained from branches grown in the laboratory and from fruit pulp collected in the field. These were desinfected and sown on Murashige-Skoog (1962 medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, benzylaminopurine (BAP and kinetin(Kin. The cultures were maintained at 25±2°C in darkness for 2 weeks and subsequently with a photoperiod of 16 hours in light and 8 hours in dark for 6 weeks. Treatment with 2 mg/L 2,4-D and 0.1 mg/L BAP allowed major callus formation in the three types of explants. Calluswere generated from the first week (knots, fourth week (leaves and sixth week (pulp and these were friable (leaves and nodes and non-friable (pulp. In conclusion, the described method is efficient for in vitro callus induction in leaves, knots and pulp of M. dubia, been leaves and knots explants more suitable for callus obtention

  14. THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE

    Ika Roostika

    2012-10-01

    Full Text Available Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid or B medium (MS + 0.018 mM benzyl adenine. The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos perexplant in 2 months. The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seedproduction.Abstrak Bahasa IndonesiaSmooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi

  15. MULTIPLICACIÓN Y TUBERIZACIÓN in vitro DE ÑAME (Dioscorea alata L. EN SISTEMA DE INMERSIÓN TEMPORAL MULTIPLICATION AND in vitro TUBERIZATION OF YAM (Dioscorea alata L. IN TEMPORARY IMMERSION SYSTEM

    Robinson Salazar Díaz

    2007-12-01

    Full Text Available Se realizó la estandarización de la multiplicación y tuberización in vitro de ñame (Dioscorea alata L. utilizando el sistema de inmersión temporal conocido como RITA®. Se evaluó inicialmente el porcentaje de brotación de segmentos nodales en medio de cultivo semisólido utilizando dos tipos de citoquininas (BAP y Kinetina. Las plantas obtenidas fueron subcultivadas cada cuatro semanas y luego utilizadas para la obtención de meristemos y plántulas para llevar a cabo los diferentes experimentos en sistema de inmersión temporal. Se obtuvo un mayor porcentaje de explantes brotados al utilizar Kinetina. Con el sistema RITA® se evaluó el efecto de la densidad de explantes sobre la tasa de multiplicación y sobre la inducción, formación y desarrollo de microtubérculos, no encontrándose diferencias estadísticamente significativas entre las densidades evaluadas. De igual manera, se evaluó el efecto de diferentes frecuencias y tiempos de inmersión sobre la tasa de multiplicación, encontrándose que a una frecuencia de inmersión cada 12 horas y durante 10 minutos se obtenía la formación de mayor número de nudos y mayor tamaño de las plantas a las cuatro semanas de cultivo. Al utilizar el sistema de inmersión temporal se alcanzaron mayores tasas de multiplicación y tuberización in vitro comparadas con el sistema de cultivo convencional. Demostrándose así, el potencial de estos sistemas para implementar un programa de producción de semillas de ñame.Multiplication rate and in vitro tuberización of yam (Dioscorea alata L. was made using the temporary immersion system RITA®. Shoot percentage of nodal segments was initially evaluated in semisolid medium using two cytokinin types ((BAP and Kinetin. The obtained plants were subcultured every four weeks and then used to obtain meristems and plantlets to carry out different experiments in temporary immersion system. A higher sprouted explants percentage was obtained when using

  16. Factors Influencing in Vitro Seed Germination, Morphogenetic Potential and Correlation of Secondary Metabolism with Tissue Development in Prunella Vulgaris L

    Fazal, H.; Shinwari, Z. K.; Abbasi, B. H.; Ahmad, N.

    2016-01-01

    Plant growth regulators (PGRs), polyamines (PAs) and temperature regimes are the key factors that influence morphogenesis and plant architectural development; however, the understanding that how these factors control plant growth and development is still poor and needs further research in Prunella vulgaris. In this study, we monitored the effect of these factors on seed germination, morphogenetic potential and secondary metabolism. Different temperature regimes showed that 25 degree C is the most suitable temperature for seed germination (88.87±1.76 percent) on Murashige and Skoog (MS) basal medium. The synergistic combinations of kinetin (Kn), 6-benzyladenine (BA) and putrescine (PUT; 2.0 mg l/sup -1/) promoted seed germination (90.22±4.51 percent) after 24 days of inoculation. A combination of Kn and PUT (1.0 mg l/sup -1/) encouraged mean shoot length (11.0±1.95 mm) with the optimum amount of chlorophyll content (23.73±1.8 micro g cm/sup -2/). However, maximum mean root length (13±0.65 mm) was observed on medium containing Kn and spermidine (SPD, 2.0 mg l-1). Maximum calli (71.56±2.63 percent) were obtained from root explants on 0.5 MS-medium containing indole butyric acid (IBA) and Alpha-naphthalene acetic acid (NAA; 0.5 mg l/sup -1/). Higher number of shoots (78.5±3.75 percent) was obtained with Kn and PUT (1.0 mg l/sup -1/). IBA concentration of 1.0 mg l/sup -1/ was found effective for root formation (74.71±3.3 percent). Moreover, PGRs and PAs have a significant effect on accumulation of total phenolics, flavonoids and DPPH activity. This protocol is helpful for consistent plantlets and prunellin production in P. vulgaris L. (author)

  17. Chemical composition of essential oil from in vitro grown peperomia obtusifolia through gc-ms

    Ilyasi, S.; Naz, S.; Aslam, F.

    2014-01-01

    Apical meristems and nodal plant parts were used for mass propagation of Peperomia obtusifolia. Different concentrations of BAP (6-benzylaminopurine), TDZ (Thidiazuron) and KIN (Kinetin) were used in MS medium. The highest shoot proliferation and multiplication formation occurred in the MS basal medium containing 1.0 mg/mL BAP. Maximum number of shoots and shoot lengths were 9.80, 0.50 and 35.40, 2.92 cm respectively. Rooting response was the best in MS basal medium fortified with 1.0 mg/mL BAP plus 0.5 mg/mL NAA (alpha-naphthalene acetic acid). For acclimatization, the rooted plantlets were transferred to the greenhouse. The volatile oil of this In vitro grown P. obtusifolia was extracted by hydro-distillation and investigated by gas chromatography/mass spectrometry (GC-MS). The chromatographic analysis of oil showed 35 constituents of which, 16 volatile compounds contributing 65.0% of the total oil constituents could be identified. The major components identified in this oil were sesquiterpenes such as caryophyllene (17.17%), apiol (16.65%), alpha-cardinol (2.12%) and alpha-caryophyllene (1.90%). The monoterpenes such as R-alpha-Pinene(1.00%), camphene (0.84%), borneal (0.32%) and limonene (0.25%) were also identified. The tau-muurolol (0.68%), gamma- Elemene (0.63%), copaene (0.43%) and tau.-cadinol (0.27%) were present comparatively in minor percentages. The caryophyllene oxide (2.95%) was oxygenated sesquiterpene, 3,5-Dimethoxy-4-hydroxycinnamic acid (0.53%) and asarone (0.41%) were phenyl-propanoids. (author)

  18. A Micropropagation Protocol for a Critically Endangered Mangrove Excoecaria Agallocha L

    Panneerselvam RAJARAM

    2012-06-01

    Full Text Available Excoecaria agallocha L. is a critically endangered mangrove tree from the Pichavaram mangrove reserve forest, the Tamil Nadu Coastal area. It is distributed on the seashore and the edge-mangrove. In order to reduce the decrease in number of these Mediterranean mixed stand, unsupervised forest management practices have drastically been reduced. In addition, the deforestation of the mangrove area, along with a low seed germination rate further endanger this species. In this study we developed a protocol for the micropropagation of adult Excoecaria agallocha. Microcuttings were obtained from lateral and apical twigs of mature plants and used as explants. Microcuttings with axillary buds were grown on different media, plant growth regulators and phenolic exudation substances. The axillary shoots produced on uncontaminated explants were excised, segmented and recultured in the same medium, to increase the stock of shoot cultures. The Modified Murashige and Skoog (MMS medium, augmented with different concentrations of N6 – benzyl adenine (BAP and Naphthalene acetic acid (NAA, either alone, or in combinations, as a potential medium for shoot multiplication by nodal segments, was tested. In the following experiment, equal molar concentrations of four cytokonins [BAP, Kinetin and 2- isopenthenyladenine (2iP] in combination with equal molar concentrations of three auxins [ NAA, Indole acetic acid (IAA and indole-3- butyric ]were used to test the rate of axillary shoot proliferation, induced on MMS agar medium supplemented with 3.9 µM BAP and 1.34 µM NAA after 6 weeks in culture. Different auxins (NAA, IBA and IAA were to determine the optimum conditions for in vitro rooting of microshoots. The best results were accomplished with NAA 5.41 µM (89% rooting and with IBA at 2.85 or 5.71µM (86% and 86.5% rooting, respectively.

  19. Micropropagation, Micromorphological Studies, and In Vitro Flowering in Rungia pectinata L.

    Shekhawat, Mahipal S; Manokari, M; Ravindran, C P

    2016-01-01

    A tissue culture protocol was developed for an important medicinal plant Rungia pectinata L. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2 solution. Murashige and Skoog (MS) medium was used to establish the cultures of R. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L(-1) 6-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L(-1) each of BAP and kinetin (Kin) + 0.1 mg L(-1) indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L(-1) indole-3 butyric acid (IBA). In vitro flowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots under in vitro conditions.

  20. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-01-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity. PMID:25183942

  1. Two-stage culture procedure using thidiazuron for efficient micropropagation of Stevia rebaudiana, an anti-diabetic medicinal herb.

    Singh, Pallavi; Dwivedi, Padmanabh

    2014-08-01

    Stevia rebaudiana Bertoni, member of Asteraceae family, has bio-active compounds stevioside and rebaudioside which taste about 300 times sweeter than sucrose. It regulates blood sugar, prevents hypertension and tooth decay as well as used in treatment of skin disorders having high medicinal values, and hence there is a need for generating the plant on large scale. We have developed an efficient micropropagation protocol on half strength Murashige and Skoog (MS) media, using two-stage culture procedures. Varying concentrations of cytokinins, i.e., benzylaminopurine, kinetin and thidiazuron (TDZ) were supplemented in the nutrient media to observe their effects on shoot development. All the cytokinins promoted shoot formation, however, best response was observed in the TDZ (0.5 mg/l). The shoots from selected induction medium were sub-cultured on the multiplication media. The media containing 0.01 mg/l TDZ produced maximum number of shoot (11.00 ± 0.40) with longer shoots (7.17 ± 0.16) and highest number of leaves (61.00 ± 1.29). Rooting response was best observed in one-fourth strength on MS media supplemented with indole-3-butyric acid (1.0 mg/l) and activated charcoal (50 mg/l) with (11.00 ± 0.40) number of roots. The plantlets thus obtained were hardened and transferred to the pots with soil and sand mixture, where the survival rate was 80 % after 2 months. Quantitative analysis of stevioside content in leaves of in vivo mother plant and in vitro plantlets was carried out by high performance liquid chromatography. A remarkable increase in stevioside content was noticed in the in vitro-raised plants as compared to in vivo grown plants. The protocol reported here might be useful in genetic improvement and high stevioside production.

  2. Micropropagation, Micromorphological Studies, and In Vitro Flowering in Rungia pectinata L.

    Mahipal S. Shekhawat

    2016-01-01

    Full Text Available A tissue culture protocol was developed for an important medicinal plant Rungia pectinata L. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2 solution. Murashige and Skoog (MS medium was used to establish the cultures of R. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L−1 6-benzylaminopurine (BAP. About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L−1 each of BAP and kinetin (Kin + 0.1 mg L−1 indole-3 acetic acid (IAA. Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length on half strength MS medium fortified with 2.0 mg L−1 indole-3 butyric acid (IBA. In vitro flowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots under in vitro conditions.

  3. Influence of Root Exudates and Soil on Attachment of Pasteuria penetrans to Meloidogyne arenaria.

    Liu, Chang; Timper, Patricia; Ji, Pingsheng; Mekete, Tesfamariam; Joseph, Soumi

    2017-09-01

    The bacterium Pasteuria penetrans is a parasite of root-knot nematodes ( Meloidogyne spp.). Endospores of P. penetrans attach to the cuticle of second-stage juveniles (J2) and subsequently sterilize infected females. When encumbered by large numbers of spores, juveniles are less mobile and their ability to infect roots is reduced. This study looked at different factors that influence spore attachment of P. penetrans to the root-knot nematode Meloidogyne arenaria . Pretreatment of J2 with root exudates of eggplant ( Solanum melongena cv. Black beauty) reduced spore attachment compared with pretreatment with phosphate-buffered saline (PBS), suggesting that the nematode surface coat was altered or the spore recognition domains on the nematode surface were blocked. Spore attachment was equally reduced following exposure to root exudates from both host and nonhost plants for M. arenaria , indicating a common signal that affects spore attachment. Although phytohormones have been shown to influence the lipophilicity of the nematode surface coat, auxins and kinetins did not affect spore attachment compared with PBS. Root exudates reduced spore attachment more in sterilized soil than in natural soil. Sterilization may have eliminated microbes that consume root exudates, or altered the chemical components of the soil solution or root exudates. Root exudates caused a greater decrease in spore attachment in loamy sand than in a sandy loam soil. The sandy loam had higher clay content than the loamy sand, which may have resulted in more adsorption of compounds in the root exudates that affect spore attachment. The components of the root exudates could have also been modified by soil type. The results of this study demonstrate that root exudates can decrease the attachment of P. penetrans endospores to root-knot nematodes, indicating that when these nematodes enter the root zone their susceptibility to spore attachment may decrease.

  4. A New Synthetic Amphiploid (AADDAA) between Gossypium hirsutum and G. arboreum Lays the Foundation for Transferring Resistances to Verticillium and Drought

    Chen, Yu; Wang, Yingying; Zhao, Ting; Yang, Jianwei; Feng, Shouli; Nazeer, Wajad; Zhang, Tianzhen; Zhou, Baoliang

    2015-01-01

    Gossypium arboreum, a cultivated cotton species (2n = 26, AA) native to Asia, possesses invaluable characteristics unavailable in the tetraploid cultivated cotton gene pool, such as resistance to pests and diseases and tolerance to abiotic stresses. However, it is quite difficult to transfer favorable traits into Upland cotton through conventional methods due to the cross-incompatibility of G. hirsutum (2n = 52, AADD) and G. arboreum. Here, we improved an embryo rescue technique to overcome the cross-incompatibility between these two parents for transferring favorable genes from G. arboreum into G. hirsutum. Our results indicate that MSB2K supplemented with 0.5 mgl-1 kinetin and 250 mg-1 casein hydrolysate is an efficient initial medium for rescuing early (3 d after pollination) hybrid embryos. Eight putative hybrids were successfully obtained, which were further verified and characterized by cytology, molecular markers and morphological analysis. The putative hybrids were subsequently treated with different concentrations of colchicine solution to double their chromosomes. The results demonstrate that four putative hybrid plants were successfully chromosome-doubled by treatment with 0.1% colchicine for 24 h and become amphiploid, which were confirmed by cytological observation, self-fertilization and backcrossing. Preliminary assessments of resistance at seedling stage indicate that the synthetic amphiploid showed highly resistant to Verticillium and drought. The synthetic amphiploid between G. hirsutum × G. arboreum would lay the foundation for developing G. arboreum-introgressed lines with the uniform genetic background of G. hirsutum acc TM-1, which would greatly enhance and simplify the mining, isolation, characterization, cloning and use of G. arboreum-specific desirable genes in future cotton breeding programs. PMID:26061996

  5. Fruit quality of tomato ‘giuliana’ treated with products with physiological effects

    Anamaria Ribeiro Pereira Ramos

    2013-12-01

    Full Text Available The present work evaluated the effect of strobilurins, boscalid, plant growth regulators and vegetal extracts on the physico-chemical quality of tomato fruits (Solanum lycopersicum L., hybrid Giuliana. The fruits from each treatment were selected and separated in 4 repetitions: control, pyraclostrobin, boscalid, pyraclostrobin + boscalid, IBA + GA3 + kinetin, GA4+7 + benzylaminopurine and vegetal extract. The first application was carried out at 30 days after transplant and the following at every 15 days. The evaluations were: weight loss, titratable acidity (TA, soluble solids (SS, SS/TA relation, pH, ascorbic acid content, texture, total soluble sugars (TSS, activity of pectin methyl esterase (PME and polygalacturonase (PG. The SS content, responsible for fruit flavor, varied accordingly to the treatment, being highest for GA4+7 + benzylaminopurine and lowest for the control. The same occurred regarding the SS/AT relation. The fruits were kept on the shelf for 9 days, at room temperature, being that at the end of this period several treatments still presented fruit in optimal consumption conditions, notably the boscalid treatment, which presented lowest weight loss, followed by pyraclostrobin. Therefore, it can be concluded that the application of the treatments did not modify the values for pH, AT and AST of the fruits. The highest PME activity was observed for the treatments with boscalid and the mixture of boscalid and pyraclostrobin, while the lowest PG activity occurred in the control and the pyraclostrobin treatment, indicating that some products accelerated the process of demethylation of pectins by PME, facilitating the action of PG.

  6. In vitro shoot regeneration and microcorm development in crocus vernus (l.) hill

    Sivanesan, I.

    2014-01-01

    An efficient method has been developed for In vitro regeneration of shoot and microcorm from corm explants of Crocus vernus. Corms were cut into 0.5-1.0 cm long segments and cultured on the SH medium supplemented with 0.5, 1.0, 2.0, or 4.0 mg L-1 2-isopentyl adenine (2-iP), N6-benzyladenine (BA), and N6-furfuryladenine (kinetin, Kin) alone or combination with 0.5 or 1.0 mg L-1 alpha-naphthalene acetic acid (NAA) for shoot regeneration. Of the three cytokinins tested, BA was found to be the most effective cytokinin for shoot formation. The number of shoots induced per explant was more when BA was combined with 0.5 mg L-1 NAA than with 1.0 mg L-1 NAA. The greatest percentage of shoot induction (97.2) with the mean number of 11.8 shoots per explant was obtained when the SH medium was supplemented with 2.0 mg L-1 BA and 0.5 mg L-1 NAA. The frequency of microcorm induction was significantly affected by the concentrations of sucrose.The greatest number of 6.1 microcorms per explant was obtained when the SH medium was supplemented with 2.0 mg L-1 BA, 0.5 mg L-1 NAA and 6.0% sucrose. The microcorms formed In vitro developed daughter corms when they were cultured on this medium. Microcorms were separated from the culture and planted out in acclimatization boxes containing a commercial medium. About 85% of corms developed shoot and root after 30 days. This protocol could be utilized for genetic transformation and mass clonal propagation of C. (author)

  7. Auxin synthesis gene tms1 driven by tuber-specific promoter alters hormonal status of transgenic potato plants and their responses to exogenous phytohormones.

    Kolachevskaya, Oksana O; Sergeeva, Lidiya I; Floková, Kristyna; Getman, Irina A; Lomin, Sergey N; Alekseeva, Valeriya V; Rukavtsova, Elena B; Buryanov, Yaroslav I; Romanov, Georgy A

    2017-03-01

    Ectopic auxin overproduction in transgenic potato leads to enhanced productivity accompanied with concerted and occasional changes in hormonal status, and causing altered response of transformants to exogenous auxin or cytokinin. Previously, we generated potato transformants expressing Agrobacterium-derived auxin synthesis gene tms1 driven by tuber-specific patatin gene promoter (B33-promoter). Here, we studied the endogenous hormonal status and the response to exogenous phytohormones in tms1 transformants cultured in vitro. Adding indole-3-acetic acid (IAA) or kinetin to culture medium affected differently tuberization of tms1-transformed and control plants, depending also on sucrose content in the medium. Exogenous phytohormones ceased to stimulate the tuber initiation in transformants at high (5-8%) sucrose concentration, while in control plants the stimulation was observed in all experimental settings. Furthermore, exogenous auxin partly inhibited the tuber initiation, and exogenous cytokinin reduced the average tuber weight in most transformants at high sucrose content. The elevated auxin level in tubers of the transformants was accompanied with a decrease in content of cytokinin bases and their ribosides in tubers and most shoots. No concerted changes in contents of abscisic, jasmonic, salicylic acids and gibberellins in tubers were detected. The data on hormonal status indicated that the enhanced productivity of tms1 transformants was due to auxin and not mediated by other phytohormones. In addition, exogenous cytokinin was shown to upregulate the expression of genes encoding orthologs of auxin receptors. Overall, the results showed that tms1 expression and local increase in IAA level in transformants affect both the balance of endogenous cytokinins and the dynamics of tuberization in response to exogenous hormones (auxin, cytokinin), the latter reaction depending also on the carbohydrate supply. We introduce a basic model for the hormonal network

  8. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    Kumar, Nitish

    2011-01-28

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  9. Inisiasi Tunas Ganda Tanaman Manggis Malinau melalui Kultur In Vitro untuk Perbanyakan Klonal

    Endang Gati Lestari

    2013-10-01

    Full Text Available Mangosteen (Garcinia mangostana L. is one of the most promising tropical fruits for export. The major constraint toincrease fruit production of the spesies is the long juvenile period. Seedless, sweet and juicy variety of mangosteen had beenfound in Malinau. In vitro propagation technique offers possibility to produce sufficient number of seedlings any time. Thisresearch was aimed at obtaining the appropriate media formula to enhance shoot proliferation. This research consisted ofshoot induction and multiplication and shoot elongation. The materials were the fresh mangosteen seeds from the Malinaumangosteen trees. The explant used in the trial was seeds which were divided into four slices. The use of 8 to 16 mg BA L-1combined with 0.2 mg thidiazuron L-1 resulted in the best shoot induction of 52 shoot buds per explant at the 6th week afterplanting with the mean height of 0.3 cm. Upon subculturing in to the similar media, the number of shoot tends to increase.For multiplication, low concentration of BA (2 to 4 mg L-1 and thidiazuron 0.05 mg L-1 were applied to increase the numbersof shoots. The total shoot number obtained in the media with 0.05 thidiazuron without BA was 11.25 and in the media with 2mg BA L-1 + 0.05 mg thidiazuron L-1 was 8.7 shoot explant-1. The result showed that the best media for shoot elongation wasMS + 1 mg BA L-1 + 2 mg kinetin L-1. The length of the shoots were in the range of 0.5-0.8 cm.Keywords: BA, Garcinia mangostana, in vitro culture, shoot multiplication, thidiazuron

  10. In vitro propagation of female Ephedra foliata Boiss. & Kotschy ex Boiss.: an endemic and threatened Gymnosperm of the Thar Desert.

    Lodha, Deepika; Rathore, Nisha; Kataria, Vinod; Shekhawat, N S

    2014-07-01

    Ephedra foliata Boiss. & Kotschy ex Boiss., (family - Ephedraceae), is an ecologically and economically important threatened Gymnosperm of the Indian Thar Desert. A method for micropropagation of E. foliata using nodal explant of mature female plant has been developed. Maximum bud-break (90 %) of the explant was obtained on MS medium supplemented with 1.5 mg l(-1) of benzyl adenine (BA) + additives. Explant produces 5.3 ± 0.40 shoots from single node with 3.25 ± 0.29 cm length. The multiplication of shoots in culture was affected by salt composition of media, types and concentrations of plant growth regulators (PGR's) and their interactions, time of transfer of the cultures. Maximum number of shoots (26.3 ± 0.82 per culture vessel) were regenerated on MS medium modified by reducing the concentration of nitrates to half supplemented with 200 mg l(-1) ammonium sulphate {(NH4) 2SO4} (MMS3) + BA (0.25 mg l(-1)), Kinetin (Kin; 0.25 mg l(-1)), Indole-3-acetic acid (IAA; 0.1 mg l(-1)) and additives. The in vitro produced shoots rooted under ex vitro on soilrite moistened with one-fourth strength of MS macro salts in screw cap bottles by treating the shoot base (s) with 500 mg l(-1) of Indole-3-butyric acid (IBA) for 5 min. The micropropagated plants were hardened in the green house. The described protocol can be applicable for (i) large scale plant production (ii) establishment of plants in natural habitat and (iii) germplasm conservation of this endemic Gymnosperm of arid regions.

  11. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity.

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-09-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity.

  12. In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers.

    Hrahsel, Lalremsiami; Basu, Adreeja; Sahoo, Lingaraj; Thangjam, Robert

    2014-02-01

    An efficient in vitro propagation method has been developed for the first time for Musa acuminata (AAA) cv. Vaibalhla, an economically important banana cultivar of Mizoram, India. Immature male flowers were used as explants. Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) were used for the regeneration process. Out of different PGR combinations, MS medium supplemented with 2 mg L(-1) 6-benzylaminopurine (BAP) + 0.5 mg L(-1) α-naphthalene acetic acid (NAA) was optimal for production of white bud-like structures (WBLS). On this medium, explants produced the highest number of buds per explant (4.30). The highest percentage (77.77) and number (3.51) of shoot formation from each explants was observed in MS medium supplemented with 2 mg L(-1) kinetin + 0.5 mg L(-1) NAA. While MS medium supplemented with a combination of 2 mg L(-1) BAP + 0.5 mg L(-1) NAA showed the maximum shoot length (14.44 cm). Rooting efficiency of the shoots was highest in the MS basal medium without any PGRs. The plantlets were hardened successfully in the greenhouse with 96% survival rate. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to assess the genetic stability of in vitro regenerated plantlets of M. acuminata (AAA) cv. Vaibalhla. Eight RAPD and 8 ISSR primers were successfully used for the analysis from the 40 RAPD and 30 ISSR primers screened initially. The amplified products were monomorphic across all the regenerated plants and were similar to the mother plant. The present standardised protocol will find application in mass production, conservation and genetic transformation studies of this commercially important banana.

  13. Rapid in vitro propagation system through shoot tip cultures of Vitex trifolia L.-an important multipurpose plant of the Pacific traditional Medicine.

    Ahmed, Rafique; Anis, Mohammad

    2014-07-01

    A rapid and efficient plant propagation system through shoot tip explants was established in Vitex trifolia L., a medicinally important plant belonging to the family Verbenaceae. Multiple shoots were induced directly on Murashige and Skoog (MS) medium consisting of different cytokinins, 6-benzyladenine (BA), kinetin (Kin) and 2-isopentenyl adenine (2-iP), BA at an optimal concentration of 5.0 μM was most effective in inducing multiple shoots where 90 % explants responded with an average shoot number (4.4±0.1) and shoot length (2.0±0.1 cm) after 6 weeks of culture. Inclusion of NAA in the culture medium along with the optimum concentration of BA promoted a higher rate of shoot multiplication and length of the shoot, where 19.2±0.3 well-grown healthy shoots with an average shoot length of 4.4±0.1 cm were obtained on completion of 12 weeks culture period. Ex vitro rooting was achieved best directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 15 min which was the most effective in inducing roots, as 95 % of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered with 92 % survival rate. The results of this study provide the first report on in vitro plant regeneration of Vitex trifolia L. using shoot tip explants.

  14. In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding.

    Patil, Jitendra Gopichand; Ahire, Mahendra Laxman; Nitnaware, Kirti Manik; Panda, Sayantan; Bhatt, Vijay P; Kishor, Polavarapu B Kavi; Nikam, Tukaram Dayaram

    2013-03-01

    Digitalis purpurea L. (Scrophulariaceae; Foxglove) is a source of cardiotonic glycosides such as digitoxin and digoxin which are commercially applied in the treatment to strengthen cardiac diffusion and to regulate heart rhythm. This investigation deals with in vitro propagation and elicited production of cardiotonic glycosides digitoxin and digoxin in shoot cultures of D. purpurea L. In vitro germinated seedlings were used as a primary source of explants. Multiple shoot formation was achieved for three explant types (nodal, internodal, and leaf) cultured on Murashige and Skoog (MS) medium with several treatments of cytokinins (6-benzyladenine-BA; kinetin-Kin; and thidiazuron-TDZ) and auxins (indole-3-acetic acid-IAA; α-naphthaleneacetic acid-NAA; and 2,4-dichlorophenoxy acetic acid-2,4-D). Maximum multiple shoots (12.7 ± 0.6) were produced from nodal explants on MS + 7.5 μM BA. Shoots were rooted in vitro on MS containing 15 μM IAA. Rooted plantlets were successfully acclimatized. To further maintain the multiple shoot induction, mother tissue was cut into four equal parts and repeatedly sub-cultured on fresh shoot induction liquid medium after each harvest. On adaptation of this strategy, an average of 18 shoots per explant could be produced. This strategy was applied for the production of biomass and glycosides digitoxin and digoxin in shoot cultures on MS medium supplemented with 7.5 μM BA and several treatments with plant growth regulators, incubation period, abiotic (salicylic acid, mannitol, sorbitol, PEG-6000, NaCl, and KCl), biotic (Aspergillus niger, Helminthosporium sp., Alternaria sp., chitin, and yeast extract) elicitors, and precursors (progesterone, cholesterol, and squalene). The treatment of KCl, mycelial mass of Helminthosporium sp., and progesterone were highly effective for the production of cardenolides. In the presence of progesterone (200 to 300 mg/l), digitoxin and digoxin accumulation was enhanced by 9.1- and 11.9-folds

  15. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    Kumar, Nitish Chandramohana

    2010-03-07

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  16. Induction and Analysis of the Alkaloid Mitragynine Content of a Mitragyna speciosa Suspension Culture System upon Elicitation and Precursor Feeding

    Nor Nahazima Mohamad Zuldin

    2013-01-01

    Full Text Available This study aimed to determine the effects of different concentrations and combinations of the phytohormones 2,4-dichlorophenoxy acetic acid (2,4-D, kinetin, 6-benzylaminopurine (BAP, and 1-naphthaleneacetic acid (NAA on callus induction and to demonstrate the role of elicitors and exogenous precursors on the production of mitragynine in a Mitragyna speciosa suspension culture. The best callus induction was achieved from petiole explants cultured on WPM that was supplemented with 4 mg L−1 2, 4-D (70.83%. Calli were transferred to liquid media and agitated on rotary shakers to establish Mitragyna speciosa cell suspension cultures. The optimum settled cell volume was achieved in the presence of WPM that contained 3 mg L−1 2,4-D and 3% sucrose (9.47±0.4667 mL. The treatment of cultures with different concentrations of yeast extract and salicylic acid for different inoculation periods revealed that the highest mitragynine content as determined by HPLC was achieved from the culture treated with 250 mg L−1 yeast extract (9.275±0.082 mg L−1 that was harvested on day 6 of culturing; salicylic acid showed low mitragynine content in all concentrations used. Tryptophan and loganin were used as exogenous precursors; the highest level of mitragynine production was achieved in cultures treated with 3 μM tryptophan and harvested at 6 days (13.226±1.98 mg L−1.

  17. Phytohormones as Important Biologically Active Molecules – Their Simple Simultaneous Detection

    Ladislav Havel

    2009-05-01

    Full Text Available Phytohormones, their functions, synthesis and effects, are of great interest. To study them in plant tissues accurate and sensitive methods are required. In the present study we aimed at optimizing experimental conditions to separate and determine not only plant hormones but also their metabolites, by liquid chromatography coupled with a UV-VIS detector. The mixture we analyzed was composed of benzyladenine, kinetin, trans-zeatin, cis-zeatin, dihydrozeatin, meta-topolin, ortho-topolin, α-naphthalene acetic acid, indole-3-acetic acid, trans-zeatin-7-glucoside, trans-zeatin-O-glucoside, trans-zeatin-9-riboside, meta-topolin-9-riboside and ortho-topolin-9-riboside. We measured the calibration dependences and estimated limits of detection and quantification under the optimal chromatographic conditions (column: Polaris C18; mobile phase: gradient starting at 2:98 (methanol:0.001% TFA and was increasing to 55:45 during twenty minutes, and then decreasing for 10 min to 35:65, flow rate: 200 µL·min-1, temperature: 50 °C, wavelength: 210 nm. The detection limits for the target molecules were estimated as tens of ng per mL. We also studied the effect of flax extracts on the phytohormones’ signals. Recovery of aliphatic and aromatic cytokinins, metabolites of cytokinins and auxinswere within the range from 87 to 105 %. The experimental conditions were tested on a mass selective detector. In addition we analysed a commercial product used for stimulation of roots formation in cuttings of poorly rooting plants. The determined content of α-naphthalene acetic acid was in good agreement with that declared by the manufacturer.

  18. Induction of Cocoa Natural Resistancy to Cocoa Pod Borer by Silica Application

    Ketut Anom Wijaya

    2009-12-01

    Full Text Available Cocoa (Theobroma cacao L. like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D with kinetin (kin. Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance

  19. Study on the presence and influence of phenolic compounds in callogenesis and somatic embryo development of cocoa (Theobroma cacao L..

    Sulistyani Pancaningtyas

    2015-03-01

    Full Text Available Cocoa (Theobroma cacao L. like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D with kinetin (kin. Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance

  20. Experimental Paper. In vitro synthesis of mucilage in Plantago ovata Forsk affected by genotypes and culture media

    Golkar Pooran

    2017-03-01

    Full Text Available Introduction: Psyllium (Plantago ovata Forsk is medicinally used mainly for its mucilage content. Objective: In the present study, an attempt was made to improve mucilage yield under in vitro callus culture using different genotypes, explants and culture media. Methods: The effects of a range of concentrations of plant growth regulators including 2,4-dichlorophenoxyacetic acid (2,4-D and kinetin (Kin were evaluated on mucilage synthesis under in vitro culture using cotyledon, hypocotyl and seed explants. Fourteen genotypes originating from different geographical regions of Iran were used to evaluate their response to in vitro mucilage synthesis. Results: The highest rate of callus induction (76% and callus growth rate CGR (0.38 mm/day were induced on MS medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l Kin and the hypocotyl explant. The results of analysis of variance showed significant genotypic differences for callus induction, CGR and mucilage content of callus and seeds. The mucilage content ranged from 0.38 to 0.08 (g/g DW and 0.13 to 0.042 (g/g DW for callus and seed, respectively. The superior callus induction (73%, CGR (0.45 mm/day and mucilage content of callus (0.38 g/g DW was denoted to Po1 genotype. The callus produced nearly three times more mucilage than the seeds using superior genotype (Po1. Conclusion: The results of this study revealed that high efficiency of callus culture of P. ovata using hypocotyl explant accompanied by the exploration of genetic diversity are important to improve the yield of mucilage synthesis by in vitro callus culture.

  1. Effects of a gamma irradiation and 5-methyltryptophan on the selection of high tryptophan accumulating rice mutants by an embryo culture system

    Kim, Dong Sub; Kim, Jin Baek; Song, Jae Young; Jeon, Jae Beom; Lee, Young Mi; Lee, Geung Joo; Kang, Si Yong [Radiation Research Center for Bio-technology, Advanced Radiation Research Institute, Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Kang, Kwon Kyoo [Division of Life Science, Hankyong National University, Anseong (Korea, Republic of); Cho, Yong Gu [College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju (Korea, Republic of); Kim, Bo Kyeong [Division of Rice Breeding and Cultivation, Honam Agricultural Research Institute, NICS, Iksan (Korea, Republic of)

    2008-11-15

    For an increase of specific free amino acids through embryo cultures, the rice (Oryza sativa L.) mutant lines resistant to a growth inhibition by 5-methyltryptophan (5MT) were selected from calli irradiated with 0-90 Gy gamma rays in 3 rice cultivars, Dongjinbyeo, Donganbyeo and Jakwangdo. The optimum 5MT concentrations for a resistance selection were 0.25 mM at the callus level. In the test of the radiation sensitivity for the callus level, radiation doses of RD50 (50% reduction on fresh weight) were 71.2, 64.0, and 68.2 Gy in the cvs. Dongjinbyeo, Donganbyeo and Jakwangdo by each exponential function. The appearance of 5MT resistant calli seemed to be dependent on a combination between the radiation dose and the 5MT selection pressure. The treatment of mutagens for the selection of amino acid analog resistant mutants has a different influence among cultivars on the resistance frequency. In the effect of growth regulators on a regeneration, a combination of 0.1 mg/l IAA and 5 mg/l kinetin, was the optimum concentration for a regeneration of calli induced from rice embryo. The regeneration rate of cv. Donganbyeo was 14.8%, which was 3.5 times and 2.6 times higher than cv. Dongjinbyeo (4.2%) and cv. Jakwangdo (5.6%), respectively. In the progeny test, the 5MT resistance character is inherited to the next generation and is expressed in the germinating M2 seeds and would appear to be a dominant trait. The 5MT resistant mutants will be useful in molecular and biochemical studies for the regulation of the nutritional quality in rice.

  2. Zeatin and Thidiazuron Induced Embryogenic Calli From In Vitro Leaf and Stem of Jojoba (Simmondsia chinensis).

    El-Ashry, Amal Abd El-Latif; Gabr, Ahmed Mohamed Magdy; Bekheet, Shawky Abd El-Hamid

    2017-01-01

    Jojoba is a promising industrial plant, which recommended with pharmaceutical benefits. The present study was conducted to stimulate embryogenic calli formation from jojoba using zeatin and thidiazuron (TDZ), as well as determination of the antioxidant activity of proliferated calli. For callus induction, leaf and stem explants derived from in vitro grown shootlets, were cultured on Murashige and Skoog (MS) medium with different combinations of 0.5 mg L-1 benzyl adenine (BA) or kinetin with 2,4-Dichlorophenoxyacetic acid (2,4-D), Naphthalene acetic acid (NAA) and picloram at 0.5 or 1mg L-1. To stimulate embryogenic calli, friable callus were transferred to woody plant medium (WPM) supplemented with different concentrations of zeatin or TDZ. Antioxidant activity of different treatments was determined using hexane or petroleum ether extraction. Data was analyzed as mean±standard deviation (SD). The MS medium supplemented with 0.5 mg L-1 BA+0.5 or 1 mg L-1 picloram was the best treatment to obtain friable calli from both explants types. WPM medium supplemented with 2 mg L-1 zeatin gave the highest percentage of embryogenic calli derived from leaf explants. While the highest percentage of embryogenic calli derived from stem explants was registered using 1 or 4 mg L-1 TDZ containing medium. Embryogenic calli originated from leaves explants on 1.5 mg L-1 zeatin showed promising activity of antioxidant with hexane extraction. However, embryogenic calli originated from stem explants on 1 mg L-1 TDZ showed the highest antioxidant activity with petroleum ether extraction. TDZ has promising effect on embryogenic callus induction from stem explants. While, zeatin has promising effect on embryogenic callus induction from leaf explants.

  3. Protocols for In Vitro Mass Multiplication and Analysis of Medicinally Important Phenolics of a Salep Orchid, Satyrium nepalense D.Don ("Salam Mishri").

    Babbar, Shashi B; Singh, Deepak K

    2016-01-01

    Satyrium nepalense is a rare and threatened medicinal orchid, populations of which in its native habitats are dwindling because of indiscriminate collections and habitat destruction, thus necessitating the development of methods for its in situ and ex situ conservation. Because of non-endospermous nature of the seeds and the immature embryos at seed dispersal stage, orchids cannot be seed-propagated as other plants. Micropropagation, using plant tissue culture techniques, offers an effective method for the multiplication of orchids. In this chapter, a five-step efficient reproducible protocol for large-scale in vitro multiplication of Satyrium nepalense is described. The first step involves asymbiotic germination of seeds isolated from immature green pods and cultured on Mitra's medium (M) gelled with 0.8 % agar and supplemented with 2 % sucrose and 1 % peptone (hereafter referred to as basal medium, BM). On this medium, seeds start germinating after a week of culture. Protocorms developed from the seeds are sub-cultured on BM fortified with 4 μM kinetin (Kn) after 8 weeks, for shoot differentiation and multiplication. The shoots developed on Kn-supplemented medium are transferred to BM alone for their elongation for the same period. The elongated shoots are transferred to the rooting medium, comprising BM supplemented with 0.5 or 1.0 μM indole-3-butyric acid, for further 8 weeks. The regenerated plantlets are transferred to a potting mix of sand and vermiculite (1:1) for acclimatization. The tubers and leaves excised from both in vitro-developed plants and those from their native habitats are analyzed and compared for the contents and concentration of medicinally important phenolics using high-performance liquid chromatography (HPLC), details of which are provided in this chapter.

  4. Asymbiotic seed germination and in vitro conservation of Coelogyne nervosa A. Rich. an endemic orchid to Western Ghats.

    Abraham, Sonia; Augustine, Jomy; Thomas, T Dennis

    2012-07-01

    Coelogyne nervosa is an epiphytic orchid endemic to Western Ghats, South India. The mature seeds of C. nervosa were cultured on ½ MS (Murashige and Skoog), MS, Kn (Knudson) and VW (Vacin and Went) media to evaluate the seed germination response. Of the four basal media used, MS medium supported maximum seed germination. Further experiments to enhance seed germination were done on MS medium supplemented with various concentrations (10, 20, 30 and 40 %) of coconut water (CW). Thirty percent CW gave the highest response in terms of percent seed germination (96), fresh weight (7.2 mg/seedling) and protocorm length (15.2 mm). Since CW containing medium did not support further seedling growth, each seedling was isolated and cultured on MS medium supplemented with either BA (6-benzylaminopurine) or Kin (kinetin) alone (1.0-4.0 mg/l each) or in combination with NAA (1-naphthaleneacetic acid; 0.2-1.0 mg/l). Maximum growth was observed on MS medium supplemented with BA (3.0 mg/l) and NAA (0.5 mg/l). On this medium, the seedlings reached an average length of 3.6 cm with 2.8 well expanded green leaves per seedling. Similarly optimum, healthy, white root induction (3.3 roots/seedlings) was also observed on the same medium. The rooted seedlings were successfully transplanted to pots with 91 % success. The 2-year-old tissue culture derived plants produced normal flowers and fruits.

  5. Premilinary studies for optimiziing a protocol for obtaining embryogenic calluses in two rubber (Hevea brasiliensis Mull. Arg clones from different geographical origins

    Santiago Cadavid Ruiz

    2006-01-01

    Full Text Available The influence of growth regulators on obtaining friable rubber (Hevea brasiliensis Müll. Arg. calluses with no plant regeneration as investigated. Two clones having different geographical origin were used in all trails carried out in this study: FX 3864 (South-American and PB 254 (Asian. Young leaves and eight- to ten-week-old seed integument from both clones were used as explants in several experiments; they were initially cultured in MH medium (Carron, et ál., 1989, modified MH medium (Montoro, et ál., 1993, 2000 and modified MS medium (Carron, et at,. 1992, no positive response being obtained by days 25 or 50. However, other trials were carried out with the integument in modified MS medium (1962, 0.67 mg/L BAP and 0.66 mg/L 2-4 D being added as medium for initiating embryogenesis, the formation of white, friable calluses being observed by day 25 in the two selected clones. These calluses were sub-cultured in MS supplemented with 0.35 mg/L BAP and 0.2 mg/L 2-4 D as callogenesis expression medium, embryogenic expression being observed in both clones by day 50. Equally friable white calluses were obtained from young leaves in the two clones in MS medium supplemented with 1.0 mg/L BAP, 1.0 mg/L ANA but without IBA and kinetin by day 25. Calluses sub-cultured in the same medium supplemented with 0.5 mg/L BAP and 0.5 mg/L ANA began to show increased friability after 50 days. culture. This work is a partial report of a macro-project for optimising a protocol for rubber (Hevea brasiliensis  multiplication by somatic embryogenesis.

  6. An efficient and reproducible method for in vitro clonal multiplication of Rauvolfia tetraphylla L. and evaluation of genetic stability using DNA-based markers.

    Faisal, Mohammad; Alatar, Abdulrahman A; Ahmad, Naseem; Anis, Mohammad; Hegazy, Ahmad K

    2012-12-01

    An efficient protocol is described for the rapid in vitro clonal propagation of an endangered medicinal plant, Rauvolfia tetraphylla L., through high frequency shoot induction from nodal explants collected from young shoots of a field grown plant. Effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) 2iP, or α-naphthalene acetic acid (NAA)], carbohydrates, different medium [Murashige and Skoog (MS), Woody Plant Medium (WPM), Gamborg medium (B5), Linsmier and Skoog medium (LS)], and various pH levels on in vitro morphogenesis were investigated. The highest frequency of shoot regeneration (90 %) and maximum number of shoot (35.4 ± 2.3) per explant were observed on WPM medium supplemented with 7.5 μM BA, 2.5 μM NAA, and 30 g/l sucrose at pH 5.8. Well-developed shoots, 4-5 cm in length, were successfully rooted ex vitro at 90 % by a 30-min pulse treatment with 150 μM IBA prior to their transfer in planting substrates. The survival rate of transplantation reached 90 % when transferred to field condition. Genetic stability of micropropagated plantlets was assessed and compared with mother plant using Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats markers. No variation was observed in DNA fingerprinting patterns among the micropropagated plants, which were similar to that of the donor plant illustrating their genetic uniformity and clonal fidelity. This confirms that clonal propagation of this plant using axillary shoot buds can be used for commercial exploitation of the selected genotype where a high degree of fidelity is an essential prerequisite. The work contributed to a better in vitro regeneration and clonal mass multiplication of R. tetraphylla and to develop a strategy for the germplasm conservation of this endangered medicinal plant.

  7. The Exogenous Amelioration Roles of Growth Regulators on Crop Plants Grow under Different Osmotic Potential

    Hamdia M. Abd El-Samad

    2014-03-01

    Full Text Available The production of fresh and dry matter of maize, wheat, cotton, broad and parsley plants show a variable response to the elevation of salinity stress. The production of fresh and dry matter of shoots and roots in wheat and broad bean plants tended to decrease with increasing NaCl concentration, salt stress progressively decrease in fresh and dry matter yield of maize plants. The increase in salinization levels induced a general insignificant change in production of fresh and dry matter of both organs of parsley plants. However, salinity induced a marked increase in the values of fresh and dry matter yields of cotton plants grown at the lowest level (-0.3 MPa NaCl and a reduction at higher salinization levels. Leaf area of unsprayed plants was excesivly decreased with the rise of osmotic stress levels especially at higher salinity levels of maize, wheat, cotton, and broad bean and parsley plants. the total pigments concentration decreased with rise of salinization levels in maize and cotton, these contents remained more or less un affected up to the level of 0.6 MPa NaCl in wheat and up to 0.9 MPa in parsley plants, there above, they were significantly reduced with increasing salinity levels. In broad bean plants the total pigments contents showed a non-significant alterations at all salinity stress. Spraying the vegetative parts of the five tested plants with 200 ppm of either GA3 or kinetin completely ameliorated the deleterious effect of salinity in fresh, dry matter, leaf area and pigment contents.

  8. Induksi Embriogenesis Somatik dari Jaringan Endosperma Jeruk Siam (Citrus nobilis Lour. cv Simadu

    Mia Kosmiatin

    2014-07-01

    Full Text Available ABSTRACTTriploid plants can be obtained from endosperm tissues through somatic embryogenesis regeneration. This research aimed to obtain somatic embryogenesis regeneration technique of tangerine endosperm. There were 3 experiments conducted in this research: 1 Embryogenic callus induction of tangerine endosperm. Endosperms isolated from fruits that were harvested from mother plants 11-13 weeks after anthesis and cultured on Murashige and Skoog (MS medium by modified vitamin Morel and Wetmore (MW which treated by 0.1 mg L-1 biotin, 500 mg L-1 malt extract (ME, 500 mg L-1 Casein hydrolisate (CH, 500 mg L-1 ME + 0.1 mg L-1 biotin, and 500 mg L-1CH + 0.1 mg L-1 biotin, 2 Maturation and germination of somatic embryos conducted by embryogenic callus cultured on MS medium by vitamin MW modified with addition of ABA, glutamine, and biotin, and 3 Plantlet elongation conducted on MS medium modified by MW vitamin with addition of GA3 and Kinetin. The best induction medium for embryogenic callus was modified MS enriched with 3 mg L-1 BA and 500 L-1 CH or ME, in succession 84.0 and 80.0%. The best medium for somatic embryos maturation with normal morphological plantlets (54.8% was modified MS medium without plant growth regulator with higher rate of solidified agent (from 2.5 to 3 g L-1 Phytagel. Plantlets elongation was highly (0.9 cm occurred on modified MS with enriched of 2.5 mg L-1 GA3. Keywords: Citrus nobilis (Lour., endosperm culture, in vitro, Simadu tangerine

  9. EFFECT OF GAMMA IRRADIATION ON THE GROWTH AND DEVELOPMENT OF SAGO PALM (Metroxylon sagu Rottb. CALLI

    Imron Riyadi

    2017-01-01

    Full Text Available The application of gamma irradiation on plant materials may increase the genetic variation of the offspring with useful traits. The experiment was conducted to determine the effect of irradiation dosage of gamma ray on growth and development of sago palm (Metroxylon sagu calli. Friable calli of sago palm derived from suspension culture were used as a material source. The primary calli were initiated from apical meristematic tissues of sago palm suckers of Alitir variety from Merauke, Papua. The treatments used were dosage of gamma ray irradiation at 0, 5, 10, 15, 20 and 25 Gy. The treated calli were then subcultured on modified Murashige and Skoog (MMS solid medium containing 3% sucrose and 0.1% activated charcoal and added with 1 mg l-1 2,4-D and 0.1 mg l-1 kinetin. The results showed that at all irradiation dosages, calli biomass increased significantly. The highest proliferation of calli biomass of 5.33 folds from the initial culture after 4 weeks was achieved at gamma irradiation of 25 Gy, whereas the lowest proliferation of calli biomass of 3.4 folds was achieved at control. The best development of embryogenic calli was obtained at 10 Gy that produced 100% somatic embryos, whereas the lowest somatic embryo formation at 0% was obtained at 0 and 25 Gy after one subculture. High response of somatic embryo induction to gamma irradiation at 10 Gy may increase production of somatic embryos. These results can be used in in vitro breeding of sago palm via mutagenesis to create new elite varieties.

  10. In vitro seed germination of economically important edible bamboo Dendrocalamus membranaceus Munro.

    Brar, Jasmine; Anand, Manju; Sood, Anil

    2013-01-01

    An in vitro propagation protocol using mature seeds of D. membranaceus was successfully established. Scarcity of seeds in bamboos because of their long flowering periods and irregular seed set resulting in low viability and germination potential, motivated us to undertake the present study. The effects of sterilants, light conditions, exogenous application of plant growth regulators and temperature in overcoming germination barriers in ageing seeds of bamboo were studied. It was found that HgCl2 (0.1%) along with bleach (15%) was more effective in raising aseptic cultures. Dark conditions, high temperatures around 30 degrees C and soaking of seeds in GA3 solution (50 ppm) overnight stimulated high percent of seed germination with corresponding increase in shoot length (2.7 +/- 0.7 mm) and number of sprouts (2.1 +/- 0.7) per explants during culture initiation. 6-benzylaminopurine acted synergistically with kinetin to give optimum germination rate of 70 +/- 13.9% as compared to 63.13% when used individually. For prolonged maintenance of cultures, 2% sucrose was found to be suitable for promoting photomixotrophic micropropagation. Following this procedure, about 65% survival of plantlets could be achieved during hardening. Biochemically seeds consume starchy endosperm for emergence of radicle which is taken as a sign of germination as also evident from the present study. Loss of viability and vigour after a year was confirmed by Tetrazolium chloride test. Micropropagation protocol developed here will ensure regeneration of large number of plants in a relatively short time. Conclusively, in vitro propagation protocol developed in D. membranaceus using mature seeds as an explants is reported for the first time.

  11. In Vitro micropropagation of Lawsonia inermis (Lythraceae

    G.R. Rout

    2001-12-01

    Full Text Available A successful protocol was developed for mass propagation of Lawsonia inermis Linn., an important medicinal plant. Multiple shoots were induced in apical and axillary meristems derived from mature explants of L. inermis on Murashige and Skoog (1962 medium supplemented with 0.25 mg/l 6-benzylaminopurine (BA, 0.25 mg/l Kinetin (Kn, 0.5 mg/l ascorbic acid and 3% (w/v sucrose. The rate of multiplication was higher when the cultures were incubated under continuous light rather than the 14h photoperiod. Rooting was readily achieved upon transferring the microshoots onto MS basal semi-solid medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA after ten days of culture. Micropropagated plantlets were acclimatized and successfully grown in soilSe desarrolo un protocolo exitoso para la propagación en masa de Lawsonia inermis Linn., una planta medicinal importante. Se indujeron múltiples tallos en meristamos apicales y auxiliares derivados de explantes maduros de L. inermis en medio de Murashige y Skoog (1962 suplementado con 0.25 mg/l 6-benzylaminopurina (BA, 0.25 mg/l quinetina (Kn, 0.5 mg/l ácido ascórbico y 3 % (w/v sucrosa. La tasa de multiplicación fue más alta cuando los cultivos fueron incubados bajo luz continua que bajo el fotoperíodo de 14hr. El enraizamiento se logró al tranferir los microtallos al medio basal, semi-sólido MS suplementado con 0.25 mg/l de ácido indol-3-butiríco (IBA luego de 10 días de cultivo. Las plántulas micropropagadas fueron aclimatizadas y cultivadas con éxito en el suelo

  12. In-vitro regeneration of sugarcane (saccharum officinarum L.)

    Azu, E.

    2009-01-01

    Sugarcane (Saccharum officinarum L.) cultivars Barbados 46 (B46), Natal Coimbatore 339 (NC0339), Ragna, local sugarcane cultivar (LSC) and Kenana (Kn) series sourced from Kyebi, Akorley, Subriso, Jejeti and the University of Ghana Agriculture Research Station, Kpong, were evaluated for sterilisation, multiplication and rooting. An efficient double sterilisation protocol was achieved by immersing axillary buds pre-treated with Goldazim (active ingredient carbendazim) in 0.2% mercuric chloride for 7 minutes followed by 0.1 % mercuric chloride for 3 minutes. At this optimal sterilisation regime, 70% of the cultured buds were decontaminated. However, post sterilisation survival required the incorporation of 0.002g/L and 0.001g/L of amphotericin Band cefotaxime respectively in the culture medium indicating that the contaminants were endophytic. Evaluation for multiple shoot induction was conducted using Murashige and Skoog (1962) basal salts (MS) medium supplemented with 25g/L sucrose, 2mg/L IAA, 2mg/L GA 3 , 3g/L activated charcoal and varying concentrations (mg/L) of BAP or kinetin. The response of the cultivars to shoot induction varied with Ragna producing the highest number of shoots (2.17) on 5mg/L BAP indicating genotypic differences. Naphthalene acetic acid (NAA) at a concentration of 5mg/L induced the highest frequency (65%) of roots in all the cultivars tested. Post-flask acclimatization and survival of plantlets was high and independent of the age at which regenerated plantlets were transferred to the plant barn for weaning. This procedure could therefore be useful for regenerating sugarcane plantlets as well as provide target tissues for genetic transformation studies (au).

  13. Short communication. N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU) enhances in vitro direct shoot organogenesis of Citrus aurantium L. epicotyl segments compared to other commonly used cytokinins

    Roussos, P. A.; Dimitriou, G.; Voloudakis, A. E.

    2011-07-01

    The effect of three concentrations of five different cytokinins, i.e. 6-benzylamino purine, 2-isopentyl adenine, kinetin (Kin), thidiazuron and N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU), was evaluated on the in vitro direct shoot organogenesis of epicotyl explants of sour orange (Citrus aurantium L.). The basal medium used was that of Murashige and Tucker and epicotyl explants were incubated in medium supplemented with the prementioned cytokinins for 45 days. The addition of Kin and 4-CPPU in the medium enhanced the direct shoot organogenesis of sour orange epicotyl segments. The concentration of each of these two cytokinins which gave the best results, was combined with indole-3-acetic acid (IAA) or {alpha}-naphthalene acetic acid ({alpha}-NAA) at concentrations ranging from 0.01 mg L{sup -}1 to 0.2 mg L{sup -}1. The inclusion of IAA at 0.2 mg L{sup -}1 in the medium with 4-CPPU at 0.05 mg L{sup -}1 resulted in 100% successful direct shoot organogenesis, while the combination of Kin at 0.25 mg L{sup -}1 with IAA or {alpha}-NAA each at 0.01 mg L{sup -}1 presented equally high organogenesis percentages (91.7%). The incubation of the produced shoots, in medium supplemented with either indole-3-butyric acid or {alpha}-NAA resulted in high rooting percentages (up to 90%) and the rooted explants were successfully acclimatized under mist (85%). Although 4-CPPU has been used in in vitro culture of various species, this is the first report on its use in the direct shoot organogenesis of citrus species and could be of great value in citrus genetic transformation protocols using epicotyl segments, since this cytokinin resulted in the absolute organogenesis percentage. (Author) 20 refs.

  14. Efficient callus formation and plant regeneration of goosegrass [Eleusine indica (L.) Gaertn.].

    Yemets, A I; Klimkina, L A; Tarassenko, L V; Blume, Y B

    2003-02-01

    Efficient methods in totipotent callus formation, cell suspension culture establishment and whole-plant regeneration have been developed for the goosegrass [ Eleusine indica (L.) Gaertn.] and its dinitroaniline-resistant biotypes. The optimum medium for inducing morphogenic calli consisted of N6 basal salts and B5 vitamins supplemented with 1-2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l(-1) glycine, 100 mg l(-1) asparagine, 100 mg l(-1) casein hydrolysate, 30 g l(-1) sucrose and 0.6% agar, pH 5.7. The presence of organogenic and embryogenic structures in these calli was histologically documented. Cell suspension cultures derived from young calli were established in a liquid medium with the same composition. Morphogenic structures of direct shoots and somatic embryos were grown into rooted plantlets on medium containing MS basal salts, B5 vitamins, 1 mg l(-1) kinetin (Kn) and 0.1 mg l(-1) indole-3-acetic acid (IAA), 3% sucrose, 0.6% agar, pH 5.7. Calli derived from the R-biotype of E. indica possessed a high resistance to trifluralin (dinitroaniline herbicide) and cross-resistance to a structurally non-related herbicide, amiprophosmethyl (phosphorothioamidate herbicide), as did the original resistant plants. Embryogenic cell suspension culture was a better source of E. indica protoplasts than callus or mesophyll tissue. The enzyme solution containing 1.5% cellulase Onozuka R-10, 0.5% driselase, 1% pectolyase Y-23, 0.5% hemicellulase and N(6) mineral salts with an additional 0.2 M KCl and 0.1 M CaCl(2) (pH 5.4-5.5) was used for protoplast isolation. The purified protoplasts were cultivated in KM8p liquid medium supplemented with 2 mg l(-1) 2,4-D and 0.2 mg l(-1) Kn.

  15. Auxin-cytokinin synergism in vitro for producing genetically stable plants of Ruta graveolens using shoot tip meristems

    Mohammad Faisal

    2018-02-01

    Full Text Available An efficient micropropagation protocol was developed for Ruta graveolens Linn. using shoot tip meristems derived from a 4-month-old field grown plant. In vitro shoot regeneration and proliferation was accomplished on Murashige and Skoogs (MS semi-solid medium in addition to different doses of cytokinins viz.6- benzyl adenine (BA, Kinetin (Kn or 2-isopetynyl adenine (2iP, singly or in combination with auxins viz. indole-3-acetic acid (IAA, indole-3-butyric acid (IBA or α-naphthalene acetic acid (NAA. Highest regeneration frequency (27.6% was obtained on (MS medium composed of BA (10 µM with maximum number (9.4 of shoots and 4.3 cm shoot length after 4 weeks of incubation. Among various combinations tried best regeneration frequency (71% of multiple shoot formation with highest number (12.6 of shoots per shoot tip explants were achieved in MS medium augmented with a combination BA (10.0 µM and NAA (2.5 µM after 4 weeks of incubation. The optimum frequency (97% of rhizogenesis was achieved on half-strength MS medium having 0.5 µM IBA after 4 weeks of incubation. Tissue culture raised plantlets with 5–7 fully opened leaves with healthy root system were successfully acclimatized off in Soilrite™ with 80% survival rate followed by transportation to normal soil under natural light. Genetic stability among in vitro raised progeny was evaluated by ISSR and RAPD markers. The entire banding pattern revealed from in vitro regenerated plants was monomorphic to the donor. The present protocol provides an alternative option for commercial propagation and fruitful setting up of genetically uniform progeny for sustainable utilization and germplasm preservation.

  16. In vitro propagation and elemental analysis of phyllanthus nirui L.: an anti-plasmodial plant

    Adusei-Fosu, K.

    2010-12-01

    Three accessions of Phyllanthus niruri from three different localities were assessed for their fruit or seed germination in vivo and in vitro as well as embryogenic calli and subsequent embryo induction. Accessions collected from Kwabenya, Kasoa or Aburi did not germinate when nursed. However, seeds from 3, 5 or 7 days dehisced fruits germinated with 7 days having the highest percentage (68.8%) of germination when nursed in the same substrate suggesting that there was fruit wall imposed dormancy. To improve percentage germination, seeds were cultured on MS medium supplemented with 0-1.2 mg/l BAP or kinetin. At this treatment, seed cultured on MS medium supplemented with 1.2 mg/l had the highest percentage (61.1% ) of germination. Alternate regeneration via nodal cuttings cultured on MS basal medium supplemented with BAP with NAA or IBA resulted in multiple shoot induction which augurs well for rapid clonal multiplication. Additionally, regeneration via somatic embryogenesis using leaf lobe explants cultured on MS, OK W, Schenk basal medium with vitamins supplemented with 2,4-0 or Picloram resulted in only callus induction. Callus induction was high when concentrations of (0.2-1.0 mg/l) 2,4-0 were used with concentrations of (2.0-3.0 mg/l) Picloram was able to induce calli. The transfer of calli to MS medium supplemented with BAP resulted in multiple shoot regeneration. Calli also grew in size and were weighed after three series of subcultures indicating that they could be used for extraction of antiplasmodial active ingredients. Instrumental Neutron Activation Analysis (lNAA) revealed the presence of five macro and four micro nutrients but no toxic element making P. niruri useful as a medicinal plant. (au)

  17. Enhanced iron and zinc accumulation in genetically engineered pineapple plants using soybean ferritin gene.

    Mhatre, Minal; Srinivas, Lingam; Ganapathi, Thumballi R

    2011-12-01

    Pineapple (Ananas comosus L. Merr., cv. "Queen") leaf bases were transformed with Agrobacterium tumefaciens strain EHA 105 harboring the pSF and pEFESF plasmids with soybean ferritin cDNA. Four to eight percent of the co-cultivated leaf bases produced multiple shoots 6 weeks after transfer to Murashige and Skoog's medium supplemented with α-naphthalene acetic acid 1.8 mg/l, indole-3-butyric acid 2.0 mg/l, kinetin 2.0 mg/l, cefotaxime 400 mg/l, and kanamycin 50 mg/l. Putatively transformed shoots (1-2 cm) were selected and multiplied on medium of the same composition and elongated shoots (5 cm) were rooted on liquid rooting medium supplemented with cefotaxime 400 mg/l and kanamycin 100 mg/l. The rooted plants were analyzed through PCR, genomic Southern analysis, and reverse transcription PCR. The results clearly confirmed the integration and expression of soybean ferritin gene in the transformed plants. Atomic absorption spectroscopic analysis carried out with six independently transformed lines of pSF and pEFE-SF revealed a maximum of 5.03-fold increase in iron and 2.44-fold increase in zinc accumulation in the leaves of pSF-transformed plants. In pEFE-SF-transformed plants, a 3.65-fold increase in iron and 2.05-fold increase in zinc levels was observed. Few of the transgenic plants were hardened in the greenhouse and are being grown to maturity to determine the enhanced iron and zinc accumulation in the fruits. To the best of our knowledge this is the first report on the transformation of pineapple with soybean ferritin for enhanced accumulation of iron and zinc content in the transgenic plants.

  18. Optimization of in vitro regeneration and Agrobacterium tumefaciens-mediated transformation with heat-resistant cDNA in Brassica oleracea subsp. italica cv. Green Marvel.

    Ravanfar, Seyed Ali; Aziz, Maheran Abdul; Saud, Halimi Mohd; Abdullah, Janna Ong

    2015-11-01

    An efficient system for shoot regeneration and Agrobacterium tumefaciens-mediated transformation of Brassica oleracea cv. Green Marvel cultivar is described. This study focuses on developing shoot regeneration from hypocotyl explants of broccoli cv. Green Marvel using thidiazuron (TDZ), zeatin, and kinetin, the optimization of factors affecting Agrobacterium-mediated transformation of the hypocotyl explants with heat-resistant cDNA, followed by the confirmation of transgenicity of the regenerants. High shoot regeneration was observed in 0.05-0.1 mg dm(-3) TDZ. TDZ at 0.1 mg dm(-3) produced among the highest percentage of shoot regeneration (96.67 %) and mean number of shoot formation (6.17). The highest percentage (13.33 %) and mean number (0.17) of putative transformant production were on hypocotyl explants subjected to preculture on shoot regeneration medium (SRM) with 200 µM acetosyringone. On optimization of bacterial density and inoculation time, the highest percentage and mean number of putative transformant production were on hypocotyl explants inoculated with a bacterial dilution of 1:5 for 30 min. Polymerase chain reaction (PCR) assay indicated a transformation efficiency of 8.33 %. The luciferase assay showed stable integration of the Arabidopsis thaliana HSP101 (AtHSP101) cDNA in the transgenic broccoli regenerants. Three out of five transgenic lines confirmed through PCR showed positive hybridization bands of the AtHSP101 cDNA through Southern blot analysis. The presence of AtHSP101 transcripts in the three transgenic broccoli lines indicated by reverse transcription-PCR (RT-PCR) confirmed the expression of the gene. In conclusion, an improved regeneration system has been established from hypocotyl explants of broccoli followed by successful transformation with AtHSP101 for resistance to high temperature.

  19. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    Kumar, Nitish Chandramohana; Vijay Anand, K. G.; Reddy, Muppala P.

    2010-01-01

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  20. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato

    Rizwan RASHID

    2010-03-01

    Full Text Available This study was conducted for developing a high frequency regeneration system in two genotypes of tomato (Lycopersicon esculentum Mill., �Punjab Upma� and �IPA-3� for direct shoot regeneration from hypocotyl explants. The explants were excised from in vitro tomato seedlings and cultured on MS medium supplemented with different concentrations and combinations of hormones. Direct regeneration was significantly influenced by the genotype hormones combination and concentrations. The MS medium supplemented with (Kinetin 0.5 mg/l and (BAP 0.5 mg/l was found optimum for inducing direct shoot regeneration and number of shoots per explant from hypocotyl explants on this medium. Shoot regeneration per cent in �Punjab Upma� and �IPA-3� per cent was recorded to be highest i.e (86.02 and (82.57 respectively. Besides this, average number shoots per explant was also highest i.e (3.16 in case of �Punjab Upma� and (2.93 in case of �IPA-3�. A significant decline was observed in percent shoot regeneration and average number of shoots per explant with increase in the hormonal concentration. Shoots were obtained and transferred to the elongation medium (MS + BAP 0.3 mg/l. Hundred per cent rooting was induced in separated shoots upon culturing on MS and � MS basal media. Hardening on moist cotton showed maximum plantlet survival rate in case of both genotypes. After hardening, plants were transferred to soil. Thus, a tissue culture base line was established in tomato for obtaining direct regeneration using hypocotyl as explants.

  1. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato

    Rizwan RASHID

    2010-03-01

    Full Text Available This study was conducted for developing a high frequency regeneration system in two genotypes of tomato (Lycopersicon esculentum Mill., Punjab Upma and IPA-3 for direct shoot regeneration from hypocotyl explants. The explants were excised from in vitro tomato seedlings and cultured on MS medium supplemented with different concentrations and combinations of hormones. Direct regeneration was significantly influenced by the genotype hormones combination and concentrations. The MS medium supplemented with (Kinetin 0.5 mg/l and (BAP 0.5 mg/l was found optimum for inducing direct shoot regeneration and number of shoots per explant from hypocotyl explants on this medium. Shoot regeneration per cent in Punjab Upma and IPA-3 per cent was recorded to be highest i.e (86.02 and (82.57 respectively. Besides this, average number shoots per explant was also highest i.e (3.16 in case of Punjab Upma and (2.93 in case of IPA-3. A significant decline was observed in percent shoot regeneration and average number of shoots per explant with increase in the hormonal concentration. Shoots were obtained and transferred to the elongation medium (MS + BAP 0.3 mg/l. Hundred per cent rooting was induced in separated shoots upon culturing on MS and MS basal media. Hardening on moist cotton showed maximum plantlet survival rate in case of both genotypes. After hardening, plants were transferred to soil. Thus, a tissue culture base line was established in tomato for obtaining direct regeneration using hypocotyl as explants.

  2. Ultraviolet-B (UV-B) radiation as an elicitor of flavonoid production in callus cultures of jatropha (Jatropha curcas L.)

    Alvero-Bascos, E.M.; Ungson, L.B.

    2012-01-01

    Callus cultures of jatropha (Jatropha curcas L.) grown in Murashige and Skoog's (MS) medium supplemented with naphthalene-acetic acid (NAA, 20 microM) and 6-furfurylaminopurine (kinetin, 20 microM) were exposed to ultraviolet-B (UV-B) radiation to investigate its potential as an abiotic elicitor of flavonoid production. Prior to irradiation, the levels of the flavonoids, apigenin, vitexin and isovitexin in the leaf and callus extracts were determined through high performance liquid chromatography (HPLC). Results showed that vitexin and isovitexin were the dominant flavonoids in the leaves while only apigenin was detected in the calli, suggesting a correlation between the degree of differentiation and biosynthesis of flavonoids in plant tissues. Irradiation of callus cultures for 7 d using two UV-B doses (12.6 and 25.3 kJ/sq m) induced synthesis of all three flavonoids (up to 780 micro g/g dw increase) to levels similar to or higher than those found in whole leaves. The combined levels of the three flavonoids in the cultures treated with the higher UV-B dose were 20-fold higher than the control and were comparable to concentrations found in leaves while a 10-fold increase in combined flavonoid levels was observed in calli irradiated with the lower UV-B dose. Furthermore, random amplified polymorphic DNA (RAPD) analyses of DNA extracts from the leaves and calli revealed that UV-B irradiation enhanced flavonoid synthesis without altering DNA sequence. These results further support the supposed involvement of UV-B in the transcriptional regulation of the expression of flavonoid biosysnthetic genes. Overall, the findings showed that elicitation through UV-B irradiation is an effective strategy to induce flavonoid production in dedifferentiated J. curcas cultures that have lost their capacity to produce the flavonoids normally synthesized in intact organs. (author)

  3. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    Kumar, Nitish; Vijay Anand, K. G.; Reddy, Muppala P.

    2011-01-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  4. Optimization of callus induction of Zataria multiflora under the effect of different plant growth regulators and explant source

    N. Mosavat*

    2017-11-01

    Full Text Available Background and objectives: The Lamiaceae family is rich in favorable secondary metabolites which have different medicinal properties and also use in food, cosmetic and sanitary industry. Zataria multiflora Boiss. is an aromatic and bushy plant containing specific pharmaceutical components which is only distributed in certain regions of Iran. Tissue culture technologies could be suitable for in vitro production of Zataria. Methods: In this study, callus production and callus related traits of Zataria was evaluated at in vitro condition. Callus induction was performed on Murashige and Skoog (MS medium containing different levels of plant growth regulators including different cytokinins (Kinetin, benzyl amino purine and auxins (2,4 dichlorophenoxyacetic acid and naphtalen acetic acid and two different explant (hypocotyl and leaf. Results: The friable calli with yellow-green color only appeared from leaf explants on three different treatments including: 1: 2.5 (mg/L 2,4-D; 2: 2 (mg/L  2,4-D; 3: [2 (mg/L 2,4-D+ 1 (mg/L Kin].  The best callus induction (75% was obtained at 2,4-D (2 mg/L + Kin (1 mg/L after 2 month of incubation under the photoperiod of 16/8 (light/dark. The highest callus growth rate (CGR (0.072 mm/day and callus fresh weight (0.135 g were denoted to the treatment of 2 mg/L (2,4-D.  Conclusion: The benefits of the protocol described here include the possibility of its use throughout the callus culture for commercial production of suitable secondary metabolites of Zataria in rapid time and huge scale.

  5. Evaluation of haemoglobin (erythrogen): for improved somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L. cv. SVPR 2).

    Ganesan, M; Jayabalan, N

    2004-10-01

    Somatic embryogenesis in cotton (Gossypium hirsutum L.) is accelerated when the plant regeneration medium is supplemented with haemoglobin (erythrogen). In cotton SVPR 2 lines, a higher frequency of embryoid formation was observed when the medium contained 400 mg/l haemoglobin. Fresh weight of the callus, rate of embryoid induction, number of embryoids formed and the percentage of plant regeneration from somatic embryos were increased. Among the two different cultivars tested, MCU 11 showed no response to the presence of haemoglobin when compared to SVPR 2, and embryogenic callus formation was completely absent in the former. Medium containing MS salts, 100 mg/l myo-inositol , 0.3 mg/l thiamine-HCL, 0.3 mg/l Picloram (PIC), 0.1 mg/l kinetin and 400 mg/l haemoglobin effected a better response with respect to embryogenic callus induction. After 8 weeks of culture, a high frequency of embryoid induction was observed on medium containing MS basal salts, 100 mg/l myo-inositol, 0.3 mg/l PIC , 0.1 mg/l isopentenyl adenine, 1.0 g/l NH4NO3 and 400 mg/l haemoglobin. Plant regeneration was observed in 75.8% of the mature somatic embryos, and whole plant regeneration was achieved within 6-7 months of culture. The regenerated plantlets were fertile and similar to in vivo-grown, seed-derived plants except that they were phenotypically smaller. A positive influence of haemoglobin was observed at concentrations up to 400 mg/l at all stages of somatic embryogenesis. The increase in the levels of antioxidant enzyme activities, for example superoxide dismutase and peroxidase, indicated the presence of excess oxygen uptake and the stressed condition of the plant tissues that arose from haemoglobin supplementation. This increased oxygen uptake and haemoglobin-mediated stress appeared to accelerate somatic embryogenesis in cotton.

  6. Evaluating the bioreducing potential of the leaves, knobs and roots of Zanthoxylum capense (small knobwood) for the synthesis of silver nanoparticles, applicable to in vitro fungal contamination control

    Bodede, Olusola; Shaik, Shakira; Govinden, Roshini; Moodley, Roshila

    2017-12-01

    In this study we report on the green synthesis of silver nanoparticles using extracts from selected morphological parts of Zanthoxylum capense. UV-vis spectra of the biosynthesised silver nanoparticles (AgNPs) revealed absorption peaks at around 450 nm, indicative of the nanoparticles’ surface plasmon resonance, whilst infrared vibrational frequencies indicated the presence of flavonoids, alkaloids, and free and bonded sugars which could be responsible for the reduction and stabilisation of the AgNPs. 1H-NMR fingerprinting of the aqueous knob extract confirmed the active bio-reducing phytochemical of the knobs to be 6-O-p-coumaroyl-β-D-glucopyranoside. The nature, shape and morphology of the biosynthesised AgNPs were examined using transmission electron microscopy (TEM), selected area electron diffraction (SAED), scanning electron microscopy (SEM) and energy dispersive x-ray (EDX) analysis. Z. capense AgNPs were mostly spherical in shape with particle sizes in the range of 4-28 nm, 7-20 nm and 4-32 nm for leaves, knobs and roots, respectively. Leaf extracts were the most efficient in the synthesis of AgNPs with an average yield of 0.027 g AgNPs per g of plant (dry mass). The AgNPs were more effective than sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) in the control of in vitro fungal contamination in nodal explants of Z. capense up to two weeks. Shoots induced from the surface sterilised explants were further used for shoot multiplication on benzyl aminopurine (BAP) and kinetin (KIN). BAP at 0.5 mg l-1 gave the highest percentage (88.6%) of explants bearing shoots with an average of 4.78 shoots per explant. A total of 15 fungal endophyte strains associated with Z. capense were identified using molecular methods.

  7. In vitro regeneration of cocona (Solanum sessiliflorum, Solanaceae) cultivars for commercial production.

    Schuelter, A R; Grunvald, A K; Amaral, A T; da Luz, C L; Luz, C L; Gonçalves, L M; Stefanello, S; Scapim, C A

    2009-08-11

    Cocona (Solanum sessiliflorum Dunal) is a solanaceous shrub native to the Amazon region that produces an edible fruit. This species has numerous advantages, particularly a high nutritional value and productivity. However, due to irregular germination and rapid loss of seed viability, there are few plantations for production on a large scale. Development of alternative propagation strategies is essential for the production of homogeneous seedlings of genotypes with superior agronomic performance. We developed techniques for in vitro regeneration of the cocona varieties Santa Luzia and Thaís for large-scale production of healthy plantlets. Twenty days after seeding, seedling segments germinated in vitro were used as explant sources. Three successive experiments were performed: one to test the effect of the explant source and combinations of two growth regulators, auxin (indole acetic acid, IAA) and kinetin (KIN), on the morphogenetic response; another to investigate the effect of the combination of growth regulators on the morphogenetic response of hypocotyl segments, and another to evaluate how sucrose concentration affects the development of adventitious shoots. The best shoot induction was obtained using hypocotyl segments and stem apices, while rhizogenesis was greatest in leaves with a petiole. The number of adventitious shoots per explant on hypocotyl segments increased with 10 and 20 mg/L KIN, combined with 0.02 mg/L IAA in the variety Santa Luzia. Sucrose combined with these growth regulator levels increased the average number of calli; these were optimally produced when 45 g/L sucrose and 0.01 mg/L IAA + 20 mg/L KIN were applied. Only sucrose concentration influenced shoot proliferation in the two S. sessiliflorum varieties, with a maximum at 17.5 g/L.

  8. Studies on the effects of mutagens on cultured cells of higher plants, (2)

    Mori, Shigeyuki; Nakanishi, Hiroo; Shiojiri, Satoshi; Murakami, Michio

    1980-01-01

    A comparative study on the effects of 60 Co gamma radiation was carried out on callus tissue, seedlings and seeds of Phaseolus vulgaris L. cv. Kentucky Wonder. Callus was obtained from the hypocotyl pieces grown on an agar-solidified, modified Linsmaier and Skoog's medium supplemented with 2, 4-D (5 mg/l), kinetin (5 mg/l) and yeast extract (1,000 mg/l). Subculture was conducted on the same medium in the dark at 28 +- 1 0 C. Callus was exposed to various doses (2.5 - 15 KR) of gamma radiation at 20 KR/hr. Fresh and dry weight, potency of growth in subculture of irradiated callus, color and friability were measured. As compared with control, irradiated callus growth showed a significant decrease with increasing doses. Callus growth was classified into three types, i.e. callus began to grow from the early stage of culture at 5 KR and below, from 6.25 - 8.75 KR there was a delayed growth, at 10 KR and over there was little visual sign of growth. The potency of growth in subculture for 28 days of irradiated- and 24-day-cultured callus was recognized at 10 KR and below. With increasing doses, color of callus darkened, and the degree of friability increased. While 10-day-old seedlings were irradiated, striking inhibition of growth occurred at 2 KR, followed by degradation of the apical meristem and growth ceased completely at 5 KR. Severe inhibition of growth of irradiated seeds occurred at 20 KR and over. A remarkable difference in radiosensitivity was observed among callus tissue, seedlings and seeds, i.e. seedlings were most sensitive, followed by callus tissue, and seeds were most resistant. (author)

  9. Plant regeneration via direct somatic embryogenesis from leaf explants of Tolumnia Louise Elmore 'Elsa'.

    Shen, Hui-Ju; Chen, Jen-Tsung; Chung, Hsiao-Hang; Chang, Wei-Chin

    2018-01-22

    Tolumnia genus (equitant Oncidium) is a group of small orchids with vivid flower color. Thousands of hybrids have been registered on Royal Horticulture Society and showed great potential for ornamental plant market. The aim of this study is to establish an efficient method for in vitro propagation. Leaf explants taken from in vitro-grown plants were used to induce direct somatic embryogenesis on a modified 1/2 MS medium supplemented with five kinds of cytokinins, 2iP, BA, kinetin, TDZ and zeatin at 0.3, 1 and 3 mg l -1 in darkness. TDZ at 3 mg l -1 gave the highest percentage of explants with somatic globular embryos after 90 days of culture. It was found that 2,4-D and light regime highly retarded direct somatic embryogenesis and showed 95-100% of explant browning. Histological observations revealed that the leaf cells divided into meristematic cells firstly, followed by somatic proembryos, and then somatic globular embryos. Eventually, somatic embryos developed a bipolar structure with the shoot apical meristem and the root meristem. Scanning electron microscopy observations showed that the direct somatic embryogenesis from leaf explants was asynchronously. The somatic embryos were found on the leaf tip, the adaxial surface and also the mesophyll through a cleft, and it reflected the heterogeneity of the explant. The 90-day-old globular embryos were detached from the parent explants and transferred onto a hormone-free 1/2 MS medium in light condition for about 1 month to obtain 1-cm-height plantlets. After another 3 months for growth, the plantlets were potted with Sphagnum moss and were acclimatized in a shaded greenhouse. After 1 month of culture, the survival rate was 100%. In this report, a protocol for efficient regenerating a Tolumnia orchid, Louise Elmore 'Elsa', was established via direct somatic embryogenesis and might reveal an alternative approach for mass propagation of Tolumnia genus in orchid industry.

  10. Polyamine patterns in haploid and diploid tobacco tissues and in vitro cultures

    Sílvia Bicudo Carone

    2010-04-01

    Full Text Available The aim of this work was to determine PAs levels in pith tissues and callus cultures from haploid and diploid tobacco plants, explanted from the apical and basal regions of the stem. These explants were cultured in an RM-64 medium supplied with IAA and kinetin, under light or in the dark, during successive subcultures. PAs levels followed a basipetal decrease in diploid and an increase in haploid, pith tissues. A similar pattern of total PAs (free + conjugated was observed for the callus of diploid and haploid plants maintained in the light, and for the haploid callus in the dark, whereas the diploid callus in the dark showed a constant increase in total PAs levels until the end of culture. The PA increase in the diploid callus in the dark was related to free Put levels increase. The ploidy status of the plants could express different PA gradients together with the plant pith and in vitro callus cultures.O objetivo deste trabalho foi determinar os níveis de PAs em tecidos de medula e cultura de calos de plantas haplóides e diplóides de tabaco, obtidas da região apical e basal do caule. Estes explantes foram cultivados em meio RM-64 suplementado com AIA e cinetina, na luz e no escuro, durante vários subcultivos. Nos tecidos medulares, os níveis de PAs apresentam um decréscimo basípeto em diplóides e um aumento em haplóides.Um padrão similar nos níveis de PAs totais (livres+ conjugadas foi observado em calos haplóides e diplóides mantidos na luz, e haplóides no escuro, enquanto os diplóides cultivados no escuro mostraram um aumento constante até o final do cultivo. O aumento no conteúdo de PAs nos calos diplóides no escuro, foi devido ao aumento do conteúdo de Put livre. Foi observado que a ploidia da planta pode expressar diferentes gradientes de PA ao longo do tecido medular e nas culturas de calos in vitro.

  11. Histologia da embriogênese somática induzida em embriões de sementes maduras de Urochloa brizantha apomítica Histology of somatic embryogenesis induced in embryos of mature seeds of the apomictic Urochloa brizantha

    Sandra Janeth Lenis-Manzano

    2010-05-01

    Full Text Available O objetivo deste trabalho foi descrever o processo de embriogênese somática em Urochloa brizantha cv. Marandu (Syn. Brachiaria brizantha cv. Marandu e fornecer subsídios para o aprimoramento dos métodos de cultura de tecidos e transformação genética. Calos embriogênicos foram obtidos por indução em embriões isolados de sementes maduras, e cultivados in vitro, em meio de cultura que continha ácido 2,4-diclorofenoxiacético, 6-benzilaminopurina e caseína hidrolisada. Plântulas foram regeneradas a partir dos calos embriogênicos, na presença de ácido naftalenoacético e cinetina. Esse processo foi descrito morfologicamente por observações em microscopia de luz de secções seriadas semifinas de tecidos fixados, ao longo do processo de regeneração, em FAA [formaldeído (40%: ácido acético glacial: etanol (50%, a 5:5:90 v/v/v]. Os embriões das sementes de U. brizantha cv. Marandu não têm epiblasto e são classificados como do tipo panicoide. Nas condições estabelecidas de cultura in vitro, calos embriogênicos e embriões somáticos de U. brizantha cv. Marandu, desenvolvem-se a partir de células meristemáticas do escutelo.The objective of this work was to describe the process of somatic embryogenesis in Urochloa brizantha cv. Marandu (Syn. Brachiaria brizantha cv. Marandu and to provide support for the improvement of tissue culture and genetic transformation methods. Embryogenic calli were obtained by induction in embryos isolated from mature seeds, and cultivated in vitro in culture medium containing 2,4-dichlorophenoxyacetic acid, 6-benzylaminopurine and hydrolyzed casein. Plantlets were regenerated from the embryogenic calli in the presence of naphthaleneacetic acid and kinetin. This process was described by morphological observations of serial semithin sections of tissues fixed along the regeneration process in FAA (40% formaldehyde: acetic acid: 50% ethanol, at 5:5:90 v/v/v, using light microscopy. Seed embryos of U

  12. Development of new varieties of chrysanthemum by mutagenesis in vitro

    Ramirez, G.Z.; Hernandez, M.A.

    2001-01-01

    Full text: At present, the industry of flower cultivation in Mexico has been demanding new varieties produced locally. There are 6,000 hectares dedicated to the cultivation of flowers for domestic use, however the export is very low. The main production area is located in Villa Guerrero, a small town near Mexico City, where 80% of the total national production is grown. In addition, approximately 10 hectares of greenhouses are dedicated to the production of flowers for export, mainly in the Peninsula de Baja California and the Altiplano Central (Central Plateau). Unfortunately, the production of flowers in Mexico has been affected by two factors: the first, stock plants must be imported from Holland, France and the United States; and the second, there are some government restrictions on their import. Due to these factors, producers are behind in recent innovations related to new varieties. An alternative to solve this problem would be meristem in vitro culture. Plantlets from two varieties 'Polaris Yellow' pompom type and 'Dramatic' margarita type, were obtained through the meristems tip culture in the MS culture medium, to which kinetin 1.0 mg/l and NAA 0.05 mg/l were added. In preliminary studies, the plant material was irradiated with doses between 10 to 60 Gy and it was possible to determine that doses higher than 35 Gy were lethal for both varieties. In this experiment, plantlets were irradiated with seven doses (7.5, 10, 15, 17.5, 20, 22.5 Gy) of 60 Co gamma rays. They were then subcultured using three types of explants: bud, leaf and internode. The best variety for production of direct organogenesis was Polaris Yellow in a range of doses between 7.5 and 15 Gy, the buds being the best explant, while the internordes and leaves were not so suitable. In contrary, the leaf was considered to be the best explant for the induction of indirect organogenesis in the variety Dramatic, in a range of doses between 10 and 20 Gy. It was possible to obtain some mutants for

  13. Optimization of Regeneration Conditions and In Vitro Propagation of Sideritis Stricta Boiss & Heldr.

    Yavuz, Dudu Özkum

    2016-09-01

    In this study the micropropagation of endemic species Sideritis stricta was investigated. Leaf segments and shoot explants (hypocotyl, single node and shoot tips) taken from in vitro growing plantlets and cultured on MS and B5 media containing different growth regulators combinations BAP (0.0, 1.0, 2.0 and 3.0mg/l) and NAA (0.0, 0.1 and 0.5mg/l). MS and B5 media supplemented with BAP (1.0, 2.0 and 3.0mg/l) and NAA (0.1mg/l) combinations or only BAP and kinetin (2.0 and 3.0mg/l) were used at the subculture experiments of shoots and MS and B5 media supplemented with different concentrations of IBA (0.0, 1.5, 3.0, 4.5 and 10.mg/l) were used at the rooting experiments. S. stricta seeds germinated at the rate of 100% when the seed coat was removed and endoperm with embryo part cultured on B5 medium. The single node explants taken from in vitro germinated and grown 30-40 days plantlets on B5 medium have been determined as the most successful explant at all used hormone combinations. B5 medium supplemented with 1.0mg/l BAP+0.1mg/l NAA and 2.0mg/l BAP+0.5mg/l NAA was determined as the most effective medium on shoot formation. At the first and second subculture, the highest shoot formation was maintained on medium supplemented with 1.0mg/l BAP+0.1mg/l NAA and the number of shoots per explant were 4 and 2.11, respectively. The highest multiplication rate has been determined as 33.76 at the end of second subculture. The best rooting was achieved on B5 medium supplemented with 4.5mg/l IBA. The rooted shoots were successfully acclimatized to outdoor conditions and survival rate was determined as 90%. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants.

    Gnanaraj, Wesely Edward; Antonisamy, Johnson Marimuthu; R B, Mohanamathi; Subramanian, Kavitha Marappampalyam

    2012-01-01

    To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants. Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration. Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of

  15. Enhanced production of phenolic acids in cell suspension culture of Salvia leriifolia Benth. using growth regulators and sucrose.

    Modarres, Masoomeh; Esmaeilzadeh Bahabadi, Sedigheh; Taghavizadeh Yazdi, Mohammad Ehsan

    2018-04-01

    Salvia leriifolia Benth. (Lamiaceae) is an endangered medicinal plant with hypoglycemic, anti-inflammatory and analgesic properties. Many of the beneficial effects of Salvia spp. are attributed to the phenolic compounds. In the present study, an efficient procedure has been developed for establishment of cell suspension culture of S. leriifolia as a strategy to obtain an in vitro phenolic acids producing cell line for the first time. The effect of growth regulators and various concentrations of sucrose have been analyzed, to optimize biomass growth and phenolic acids production. The callus used for this purpose was obtained from leaves of 15-day-old in vitro seedlings, on Murashige and Skoog (MS) basal medium supplemented with different hormone balances including benzylaminopurine (BAP) and indole butyric acid (IBA); 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KIN); naphthaleneacetic acid (NAA) and BAP. Modified MS medium supplemented with 5 mg/L BAP and 5 mg/L NAA was the optimal condition for callus formation with the highest induction rate (100%), the best callus growth and the highest phenolic acids content. No callus induction was observed in combinations of IBA and BAP. Cell suspension cultures were established by transferring 0.5 g of callus to 30 mL liquid MS medium supplemented with 5 mg/L BAP and 5 mg/L NAA. Dynamics of phenolic acids production has been investigated during the growth cycle of the suspension cultures. The maximum content of caffeic acid and salvianolic acid B were observed on the 15th day of the cultivation cycle while the highest amount of rosmarinic acid was observed on the first day. In response to various sucrose concentrations, cell cultures with 40 g/L sucrose not only produced the highest dry biomass but also the highest induction of caffeic acid and salvianolic acid B. The highest amount of rosmarinic acid was observed in media containing 50 g/L sucrose. These prepared cell suspension cultures provided a useful

  16. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090 Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090

    Regina Aparecida Correia Gonçalves

    2008-03-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDSThe leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  17. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni = Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni

    Ana Paula Patrão

    2007-07-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDS.The leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  18. DIRECT AND INDIRECT SOMATIC EMBRYOGENESIS ON ARABICA COFFEE (Coffea arabica

    Meynarti Sari Dewi Ibrahim

    2013-10-01

    Full Text Available Propagation of Coffea arabica L. through direct and indirect somatic embryogenesis technique is promising for producing large number of coffee seedlings. The objectives of the research were to evaluate methods for direct and indirect somatic embryo-genesis induction of C. arabica var. Kartika. The explants were the youngest fully expanded leaves of arabica coffee. The evalu-ated medium was modified Murashige and Skoog (MS medium supplemented with a combination of 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; 4.52 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; or 9.04 µM 2,4-D + 9.08 µM thidiazuron. Both calli (100 mg and pre-embryos developed from the edge of leaf explants were subcultured into regeneration medium (half strength MS with modified vitamin, supplemented with kinetine 9.30 µM and adenine sulfate 40 mg L-1. The results showed coffee leaf explant cultured on medium containing 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron to induce direct somatic embriogenesis from explant, while that of 4.52 or 9.04 µM 2,4-D + 9.08 µM thidiazuron to induced indirect somatic embrio-genesis. The medium for calli induction from coffee by explants was medium supplemented with 4.52 or 9.04 µM 2,4-D in combination with 9.08 µM thidiazuron. On the other hand, the best medium for activation of induction of somatic embryos was MS medium supplemented with 9.04 µM 2,4-D + 9.08 µM thidiazuron. Based on this results, the first step for developing micropropagation for coffee has been resolved. The subsequent studies will be directed to evaluate agronomic performance of the derived planting materials.

  19. MORPHOLOGICAL CHANGES DURING THE DEVELOPMENT OF SOMATIC EMBRYOS OF SAGO (Metroxylon sagu Rottb.

    Pauline D. Kasi

    2016-10-01

    Full Text Available Development of somatic embryos of sago (Metroxylon sagu Rottb. on agar-solidified medium are highly varied producing heterogeneous seedlings. Understanding of this phenomenon may help in improving the cultural procedures and conditions of sagosomatic embryogenesis to obtain uniform seedlings in a large scale. This experiment was conducted at the laboratory for plant cell culture and micropropagation, Indonesian Biotechnology Research Institute for Estate Crops from January to March 2006 to examine morphological changes i.e. color and development stages of sago during their somatic embryo development on an agar-solidified medium. Twenty single globular somatic embryos of sago with specific color (yellowish, greenish, and reddish were cultured in a Petri dish supplemented with a solid medium. The medium was a micronutrients-modified MS (MMS with half strength of macronutrients containing 0.01 mg l-1 ABA, 2 mg l-1 kinetin, 20 g l-1 sucrose, 0.5 g l-1 activated charcoal, and 2 g l-1 gelrite. Parameter observed was the percentage of embryo’s number based on color and developmental stage. The result showed that at the end of 6-week culture passage, most originally greenish (80.8% and reddish (95.8% embryos remained unchanged in their colors, whereas almost half of the originally yellowish embryos turned to greenish and only 30%remained yellowish. At the same time, single globular embryos have changed gradually into the next developmental stages, although not all of the embryos were germinated. The initial color of embryo affected the rate of the developmental stage changes. Yellowish and greenish globular embryos developed more rapidly into cotyledon or germinant stages at 58% and 55% respectively, in 6 weeks than the reddish ones (41%. Therefore, the yellowish and greenish embryos are the best sources of material for in vitro mass propagation and synthetic seed production of sago.

  20. Cloning and expression of 1-aminocyclopropane-1-carboxylate oxidase cDNA induced by thidiazuron during somatic embryogenesis of alfalfa (Medicago sativa).

    Feng, Bi-Hong; Wu, Bei; Zhang, Chun-Rong; Huang, Xia; Chen, Yun-Feng; Huang, Xue-Lin

    2012-01-15

    Embryogenic callus (EC) induced from petioles of alfalfa (Medicago sativa L. cv. Jinnan) on B5h medium turned green, compact and non-embryogenic when the kinetin (KN) in the medium was replaced partially or completely by thidiazuron (TDZ). The application of CoCl₂, which is an inhibitor of 1-aminocyclopropane-1-carboxylate oxidase (ACO), counteracted the effect of TDZ. Ethylene has been shown to be involved in the modulation of TDZ-induced morphogenesis responses. However, very little is known about the genes involved in ethylene formation during somatic embryogenesis (SE). To investigate whether ethylene mediated by ACO is involved in the effect of TDZ on inhibition of embryogenic competence of the alfalfa callus. In this study we cloned full-length ACO cDNA from the alfalfa callus, named MsACO, and observed changes in this gene expression during callus formation and induction of SE under treatment with TDZ or TDZ plus CoCl₂. RNA blot analysis showed that during the EC subcultural period, the expression level of MsACO in EC was significantly increased on the 2nd day, rose to the highest level on the 8th day and remained at this high level until the 21st day. However, the ACO expression in the TDZ (0.93 μM)-treated callus was higher than in the EC especially on the 8th day. Moreover the ACO expression level increased with increasing TDZ concentration during the subcultural/maintenance period of the callus. It is worth noting that comparing the treatment with TDZ alone, the treatment with 0.93 μM TDZ plus 50 μM CoCl₂ reduced both of the ACO gene expressions and ACO activity in the treated callus. These results indicate that the effect of TDZ could be counteracted by CoCl₂ either on the ACO gene expression level or ACO activity. Thus, a TDZ inhibitory effect on embryogenic competence of alfalfa callus could be mediated by ACO gene expression. Crown Copyright © 2011. Published by Elsevier GmbH. All rights reserved.

  1. Indole-3-butyric acid promotes adventitious rooting in Arabidopsis thaliana thin cell layers by conversion into indole-3-acetic acid and stimulation of anthranilate synthase activity.

    Fattorini, L; Veloccia, A; Della Rovere, F; D'Angeli, S; Falasca, G; Altamura, M M

    2017-07-11

    Indole-3-acetic acid (IAA), and its precursor indole-3-butyric acid (IBA), control adventitious root (AR) formation in planta. Adventitious roots are also crucial for propagation via cuttings. However, IBA role(s) is/are still far to be elucidated. In Arabidopsis thaliana stem cuttings, 10 μM IBA is more AR-inductive than 10 μM IAA, and, in thin cell layers (TCLs), IBA induces ARs when combined with 0.1 μM kinetin (Kin). It is unknown whether arabidopsis TCLs produce ARs under IBA alone (10 μM) or IAA alone (10 μM), and whether they contain endogenous IAA/IBA at culture onset, possibly interfering with the exogenous IBA/IAA input. Moreover, it is unknown whether an IBA-to-IAA conversion is active in TCLs, and positively affects AR formation, possibly through the activity of the nitric oxide (NO) deriving from the conversion process. Revealed undetectable levels of both auxins at culture onset, showing that arabidopsis TCLs were optimal for investigating AR-formation under the total control of exogenous auxins. The AR-response of TCLs from various ecotypes, transgenic lines and knockout mutants was analyzed under different treatments. It was shown that ARs are better induced by IBA than IAA and IBA + Kin. IBA induced IAA-efflux (PIN1) and IAA-influx (AUX1/LAX3) genes, IAA-influx carriers activities, and expression of ANTHRANILATE SYNTHASE -alpha1 (ASA1), a gene involved in IAA-biosynthesis. ASA1 and ANTHRANILATE SYNTHASE -beta1 (ASB1), the other subunit of the same enzyme, positively affected AR-formation in the presence of exogenous IBA, because the AR-response in the TCLs of their mutant wei2wei7 was highly reduced. The AR-response of IBA-treated TCLs from ech2ibr10 mutant, blocked into IBA-to-IAA-conversion, was also strongly reduced. Nitric oxide, an IAA downstream signal and a by-product of IBA-to-IAA conversion, was early detected in IAA- and IBA-treated TCLs, but at higher levels in the latter explants. Altogether, results showed that IBA induced

  2. In vitro propagation, micromorphological studies and ex vitro rooting of cannon ball tree (Couroupita guianensis aubl.): a multipurpose threatened species.

    Shekhawat, Mahipal S; Manokari, M

    2016-01-01

    In vitro propagation methods using seeds and nodal segments of a 21-year old Couroupita guianensis - a medicinally important but threatened tree have been developed. Hundred percent of the seeds germinated on half strength Murashige and Skoog (MS) medium with 2.0 mg l(-1) indole-3 butyric acid (IBA). Nodal segments were found most suitable for the establishment of cultures. About 90 % explants responded and 4.1 ± 0.23 shoots per node were induced after five weeks of inoculation on MS medium +4.0 mg l(-1) 6-benzylaminopurine (BAP). Further shoot multiplication was achieved by repeated transfer of mother explants and subculturing of in vitro produced shoots on fresh medium. Maximum number (8.2 ± 0.17) of shoots were regenerated on MS medium with 1.0 mg l(-1) each of BAP and Kinetin (Kin) + 0.5 mg l(-1) α-naphthalene acetic acid (NAA) with additives (50 mg l(-1) of ascorbic acid and 25 mg l(-1) each of adenine sulphate, L-arginine and citric acid). The multiplied shoots rooted (4.3 ± 0.26 roots/shoot) on half strength MS medium with 2.5 mg l(-1) IBA. All the shoots were rooted ex vitro when pulse treated with 400 mg l(-1) of IBA for five min with an average of 7.3 ± 0.23 roots per shoot. Nearly 86 % of these plantlets were acclimatized within 7-8 weeks and successfully transferred in the field. Biologically significant developmental changes were observed during acclimation particularly in leaf micromorphology in terms of changes in stomata, veins and vein-islets, and trichomes. This study helps in understanding the response by the plants towards outer environmental conditions during acclimatization. This is the first report on micropropagation of C. guianensis, which could be used for the large-scale multiplication, restoration and conservation of germplasm of this threatened and medicinally important tree.

  3. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    Kumar, Nitish; Vijay Anand, K.G.; Pamidimarri, D.V.N. Sudheer; Sarkar, Tanmoy; Reddy, Muppala P.; Radhakrishnan, T.; Kaul, Tanushri; Reddy, M.K.; Sopori, Sudhir K.

    2010-01-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  4. Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration

    Sharma, Sweta K.; Kumar, Nitish Chandramohana; Reddy, Muppala P.

    2011-01-01

    Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5mgL-1 TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca2+ concentrations to 0.22gL-1 in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2mgL-1 Kn (Kinetin) and 1mgL-1 BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5mgL-1 IAA (indole-3-acetic acid) and 0.5mgL-1 BAP and 3.01-3.91cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3mgL-1 IBA (indole-3-butyric acid), 1mgL-1 IAA, 1mgL-1 NAA (α-naphthalene acetic acid) and subsequent transfer on 0

  5. In vitro plant regeneration of Albizia lebbeck (L. from seed explants

    S. Perveen

    2013-07-01

    lebbeck.Keywords: Woody legume; TDZ; multiplication; epicotyl segment; shoot stumps.Abbreviations used: GA3 (Gibberellic acid; BA (6-Benzyladenine; Kn (Kinetin; TDZ (Thidiazuron; MS (Murashige and Skoog Medium; IBA (Indole-3-butyric acid; NAA (α-Naphthalene acetic acid.

  6. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    Kumar, Nitish

    2010-07-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  7. Rapid in vitro multiplication and restoration of Celastrus paniculatus Willd. sub sp. paniculatus (Celastraceae), a medicinal woody climber.

    Nair, L G; Seeni, S

    2001-07-01

    Nodes, shoot tips, internodes and leaf bases (approximately 1.0 cm) excised from young vines of the flowering woody climber, Celastrus paniculatus WilId. sub. sp. paniculatus (Celastraceae) were cultured in Murashige and Skoog (MS) medium containing agar (0.6%), sucrose (3%) and varied concentrations of 6-benzyl aminopurine (BAP) and kinetin. All the explant types were regenerative and maximum number (3.6) and frequency (94%) of axillary shoot formation of (5.08 cm long) was recorded in the nodes cultured in BAP (1 mg L(-1)) after 6 weeks. Combinations of BAP (1 mg L(-1)) and indole-3-acetic acid/l-naphthalene acetic acid (0.01-1 mg L(-1); IAA/NAA) tested with nodes induced formation of less number (3 and 2.2) of shoots at same frequency (94%). All the explant types viz. node, shoot tip, internode and leaf base of in vitro derived shoots responded earlier and better in lower concentrations of BAP (0.5-2 mg L(-1)) with formation of 8, 3.1, 6.4 and 1.8 shoots respectively during the same period. In spite of the advanced and increased caulogenic responses, differences in cytokinin requirements between different explants observed during culture initiation still persisted with the nodes, shoot tips, internodes and petiole segments responding best at 0.5, 1 and 2 mg L(-1) BAP, respectively. The repeated reculture up to 10 cycles of the nodes from the shoot cultures each at 6-week intervals enabled multiplication and stocking of shoots without decline. Rooting of 3-7 cm shoot cuttings was induced in half-strength MS liquid medium containing IAA (1 mg L(-1)) with formation of 7.25 roots of 2.41 cm length within 6 weeks. Rooted plants were established at 84-96% rate in community pots without hardening, the least value (84%) being obtained with NAA- induced thick and calloid rooted plants. Four month old community potted plants were reintroduced into native forest habitats at 95% efficiency and 8 months after restoration, the plants were uniform in morphological, growth

  8. Olfactory stem cells, a new cellular model for studying molecular mechanisms underlying familial dysautonomia.

    Nathalie Boone

    Full Text Available BACKGROUND: Familial dysautonomia (FD is a hereditary neuropathy caused by mutations in the IKBKAP gene, the most common of which results in variable tissue-specific mRNA splicing with skipping of exon 20. Defective splicing is especially severe in nervous tissue, leading to incomplete development and progressive degeneration of sensory and autonomic neurons. The specificity of neuron loss in FD is poorly understood due to the lack of an appropriate model system. To better understand and modelize the molecular mechanisms of IKBKAP mRNA splicing, we collected human olfactory ecto-mesenchymal stem cells (hOE-MSC from FD patients. hOE-MSCs have a pluripotent ability to differentiate into various cell lineages, including neurons and glial cells. METHODOLOGY/PRINCIPAL FINDINGS: We confirmed IKBKAP mRNA alternative splicing in FD hOE-MSCs and identified 2 novel spliced isoforms also present in control cells. We observed a significant lower expression of both IKBKAP transcript and IKAP/hELP1 protein in FD cells resulting from the degradation of the transcript isoform skipping exon 20. We localized IKAP/hELP1 in different cell compartments, including the nucleus, which supports multiple roles for that protein. We also investigated cellular pathways altered in FD, at the genome-wide level, and confirmed that cell migration and cytoskeleton reorganization were among the processes altered in FD. Indeed, FD hOE-MSCs exhibit impaired migration compared to control cells. Moreover, we showed that kinetin improved exon 20 inclusion and restores a normal level of IKAP/hELP1 in FD hOE-MSCs. Furthermore, we were able to modify the IKBKAP splicing ratio in FD hOE-MSCs, increasing or reducing the WT (exon 20 inclusion:MU (exon 20 skipping ratio respectively, either by producing free-floating spheres, or by inducing cells into neural differentiation. CONCLUSIONS/SIGNIFICANCE: hOE-MSCs isolated from FD patients represent a new approach for modeling FD to better

  9. Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration

    Sharma, Sweta K.

    2011-07-01

    Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5mgL-1 TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca2+ concentrations to 0.22gL-1 in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2mgL-1 Kn (Kinetin) and 1mgL-1 BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5mgL-1 IAA (indole-3-acetic acid) and 0.5mgL-1 BAP and 3.01-3.91cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3mgL-1 IBA (indole-3-butyric acid), 1mgL-1 IAA, 1mgL-1 NAA (α-naphthalene acetic acid) and subsequent transfer on 0

  10. Assessment of Direct Regeneration in Germany (Matricaria chamomilla L. and Shirazi Chamomiles (Matricaria recutita L.

    A. Masoumiasl

    2016-07-01

    Full Text Available Introduction: German (Matricaria chamomilla L. and Shirazi (Matricaria recutita L. chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, Kohgiluyeh and Boyerahmad, Golestan and Hamedan provinces. In vitro propagation of plants have higher potential to produce qualified natural products, restoring and preserving of endangered plants, induction of somaclonal variation, industrial reproduction, valuable secondary metabolites and increased active ingredients. Researchers reported successful micropropagation system for five chamomile varieties on MS medium contained 0.01 mg/l NAA and 2.5 mg/l kinetin. The aim of this study was to investigate the effects of stem (with and without node, leaf and cotyledon explants and different plant growth regulators on direct regeneration of German and Shirazi chamomiles. Although the Shirazi chamomile is native to Iran, but German chamomile is Iran non-indigenous cultivar. By our knowledge, there have been no comparison reports about responses of these cultivars to tissue culture. Materials and Methods: This research was performed in the central laboratory of Agriculture Faculty in Yasouj University. Seeds were provided from Pakan-Bazr institute, Isfahan. Chamomile seeds were disinfected by ethanol (70% for 5-10 min and sodium hypocholorite 3% for 5-12 min and then washed for several times by distilled water. Then, seeds were sown on MS medium for germination. After 2-3 weeks, seedlings were grown and then planted in MS medium supplemented with hormonal combinations of NAA at two levels (0.1 and 0.5 mg/l, kinetin at three levels (2, 2.5 and 3 mg/l and Zeatin, BAP and 2ip at three levels (0.5, 1 and 1.5 mg/l. The experiment was performed in a completely randomized design with four replications. Factors included explants, cultivars and hormonal

  11. Micropropagação do porta-enxerto de videira '420-A' Micropropagation of '420-A' grapevine rootstock

    Pryscilla Menarin Dzazio

    2002-12-01

    meio MS/2 e aclimatização em substrato vermiculita ou Plantmax®.The objective of this work was to establish a protocol for the rootstock of 420-A micropropagation. The establishment of the cultures was accomplished with nodal segments, whose source of explants was the budding of woody stakes stored under refrigeration. In the initial cultivation were tested: the effect of 6-benzilaminopurine and kinetin with 0, 1, 5 and 10 µM concentrations, different culture medium (MS, NN and WPM and dilutions of the basic medium (MS, MS/2, MS/4 and MS/8. In the alongation and multiplication steps were tested the following culture medium: MS, MS/2, NN and WPM. In the rooting were tested: the MS/2 culture medium with or without activated coal (1gL-1. In the acclimatization were tested the substrate vermiculite, Plantmax and carbonized rice hulls. The kinetin didn't present effect on the budding and the growth of the nodal segments. BAP already promoted an increase in the number of sprouts per explant. The increase in the concentration of BAP reduced the number of leaves emitted by explant and increased the vitrification symptoms, being the best results obtained with 1 µM of BAP. In the initial cultivation the MS medium culture with the normal concentration of salts allowed the largest growth of the buddings. The dilutions of the MS medium in ¼ and 1/8 showed very harmful to the development of the rootstock '420-A', being quite harmed the growth of the shoots after the first subcultivation. During the multiplication the medium MS/2 showed more appropriate. The roots happened naturally during the multiplication, being unnecessary the use of activated coal in the culture medium. The acclimatization was accomplished with success in a misty camera, being obtained high survival rates in vermiculite (95,8% and Plantmax® (87%. It is concluded that the rootstock '420-A' can be micropropagated by initial cultivation of nodal segments in a culture medium of MS+1µM of BAP, multiplication

  12. Efecto de dos citoquininas, ácido ascórbico y sacarosa en la obtención de plantas in vitro de Sorghum bicolor para la formación de callos

    Silvio de Jesús Martínez Medina

    2012-07-01

    grown under controlled conditions, which were disinfected and were placed in a germination medium with MS salts, myo-inositol 100 mg L-1, 3% sucrose, pH 5.7, phytagel 2.5 g L-1. Different concentrations of 6-benzylaminopurine (6-BAP and kinetin were studied for shoot multiplication. Using 6 - BAP 0.22 mg L-1 in the multiplication culture medium increased the number of shoots per explant, showing phenolic oxidation. Adding 50 mg L-1 of ascorbic acid to the culture medium resisted phenolic oxidation and the number of sprouts (5.0, height and number of leaves increased significantly. Quality in vitro buds for callus formation were obtained  stimulating the thickening by using MS salts, without growth regulators, ascorbic acid 50 mg L-1, phytagel 2.5 g L-1 and pH 5.7 and 40 g L-1 of sucrose. The largest diameter of shoots, height, number of leaves and roots of in vitro plants, was achieved after 21 days of cultured in MS salts with 40 g L-1 sucrose. Calluses formation with embryogenic structures from the central cylinder of curled leaves of the nearest segment to the base of thickened shoots was achieved after 45 days of culture. Segments from the central cylinder of the curled leaves are a safe source for the formation of embryogenic calli. Key words: 6-BAP; in vitro shoots; callus with embryogenic structures; kinetin

  13. Method of propagation, stimulation of rhizomes growth in vitro culture and adaptation in the open ground for the genus Miscanthus representatives

    С. М. Гонтаренко

    2017-09-01

    Full Text Available Purpose. To develop a method of propagation, stimulation of rhizomes growth in vitro culture for the genus Miscanthus representatives and their adaptation in the open field without the use of greenhouse complexes for acclimatization and completion of growing. Methods. Biotechnological procedures, mathematical and statistical analyses. Results. Prescription of nutrient medium was developed for explants inoculation, sprouts propagation, rhizomes growth stimulation in vitro. Such sterile explants as seeds, buds to be removed from rhizomes, parts of  stems with bud were placed on modified media with mineral portion by Murashige and Skoog (MS that contained 0,5–1 dose of macroelements and one dose of microelements,  vitamins (10 mg/l of thia­minum, 1,0 mg/l of pyridoxine, 1,0 mg/l of nicotinic acid and 1,0 mg/l of ascorbic acid supplemented with amino acids (250 mg/l of glutamic acid, 3 mg/l of tyrosine, 3 mg/l of arginine, 2 mg/l of hydroxyproline, plant growth regulators [0,5–1,0 mg/l of GA (gibberelline acid, 0,2 mg/l of 6-BAP (6-Benzylaminopurine, 0,1 mg/l of NAA (α-naphtylacetic acid] in different variations. After seed germination, buds emerging and sprouts formation 1–2 cm in height, for propagation purpose they were passivated on the medium of other composition that differed from previous one by the content and ratio of growth regulators, especially by a high concentration of cytokinins [6-BAP (0,4–0,5 mg/l, kinetin (0,5 mg/l, adenine (0,5 mg/k] in different variations in presence of GA (0,2 mg/l. In order to stimulate rhizomes growth, microclones were transferred on media with other composition and ratio growth regulators (6-BAP (0,2–0,3 mg/l + GA (0,5–1,0 mg/l or 6-BAP (0,2–0,3 mg/l + GA (0,5–1,0 mg/l + NAA (0,1 mg/l,  in other words,  with a high content of gibberellins. After the formation of rhizomes 10–15 cm in length, miscanthus plants were planted out in the open ground. Stimulation of

  14. In vitro mutagenesis for development of novel variants in Rose (Rosa X Hybrida)

    Namita, B.; Raju, D.V.S.; Prasad, K.V.; Singh, Kanwar P.

    2014-01-01

    Rose (Rosa x hybrida) is the top ranking cut flower in international market due to its attractive flowers with longer stem and fragrance. Modern roses are the result of hybridization, selection and spontaneous mutations. In floriculture industry, the demand of the novel varieties of rose is increasing day by day due to change in trends and taste. Mutation breeding is an already established method for improvement of flower crops. It offers several advantages over conventional methods for the improvement of one or more traits within a short span of time. Induced mutation is one of the potent methods to induce variability in existing rose genotypes. Mutation breeding combined with tissue culture i.e. in vitro mutagenesis has proven more effective in rose due to controlled environment that provides ideal conditions for survival of mutated tissues or cells. Keeping these aspects in view, the present investigation was carried out to induce variability in rose cv. Grand Gala under in vitro conditions using gamma (γ) rays. Single node cuttings were excised from the field grown plants and were irradiated with different doses of γ rays (0,10, 20, 30, 40, 50, 60, 70 and 80 Gy) using a 60 Co source. The γ-irradiated explants were then pretreated, surface sterilized and cultured aseptically on Murashige and Skoog basal medium supplemented with 2.5 mg 5-benzylaminopurine (BAP) plus 5.0 mg kinetin plus 0.1 mg a-naphthaleneacetic acid (NAA) plus 0.5 mg gibberellic acid (GA 3 ) plus 40 mg adenine sulphate plus 0.8% (w/v) agar-agar to induce sprouting and shoot proliferation. Explants treated at higher dose of γ-rays (70 and 80 Gy) showed deleterious effects of ionising radiation. The 30 Gy treatment was found to be the LD 50 dose. It was observed that few explants treated with γ-rays sprouted, showed slow growth and failed to survive after the first sub culture. The explants irradiated with 50 Gy γ-rays exhibited minimum explant survival, bud sprouting, number of shoots

  15. Tratamento de matrizes de cravo (Dianthus caryophyllus L., Caryophyllaceae com nitrogênio e calogênese in vitro Nitrogen treatment of carnation (Dianthus caryophyllus l., Caryophyllaceae and in vitro calogenesis

    Telma Kazumi Hayashi

    2002-03-01

    culture media containing Murashige & Skoog (1962 salts and vitamins, supplemented with 1 g L-1 hidrolysed casein, 2 mumol L-1 kinetin and 3 mumol L-1 2,4-D. No plant deficiency and toxicity symptoms were apparent on the treated plants during the 60 day treatment. The nitrogen treatment affected calogenesis in relation to calli fresh and dry weights. Callus fresh weigth yield was proportional to nitrogen concentration up to 267 mg L-1 for leaf explant during 30 days. Longer treatments (45 and 60 days affected calogenesis negatively which were inversely proportional to the nitrogen concentration of the nutrient solution.

  16. Crescimento de plantas de Salvia officinalis sob ação de reguladores de crescimento vegetal Growth of Salvia officinalis plants under action of plant growth regulators

    Juliana Aparecida Povh

    2008-11-01

    consisted of the spraying of the solution of 100mg L-1 of gibberellic acid (GA3; 100mg L-1 of benzylaminopurine (BAP; 100mg L-1 of 2-chloroethyl phosphonic acid (ethephon; Stimulate® at 2% (90mg L-1 of kinetin, 50mg L-1 of gibberellic acid and 50mg L-1 of indolylbutyric acid and water (control. Applications of plant growth regulators were performed in 3 times, at 15, 25, and 35 days after transplanting, and growth was evaluated in five successive harvests at 21-day intervals, performed at 47, 68, 89, 110, and 131 days after transplanting. The following growth index were determined: leaf area ratio (LAR, specific leaf area (SLA, net assimilation rate (NAR, and relative growth rate (RGR. These results suggest that the plant growth regulators influence the index growth analysis. The plants treated with BAP presented increase values of LAR to the 47 days after transplanting. The plants treated with GA3 to NAR presented increase until the 131 days after transplanting. The RGR is decreasing for all the treatments with vegetal regulators tested and the control.

  17. Stock indexing and Potato virus Y elimination from potato plants cultivated in vitro Indexação de matrizes e eliminação do Potato virus Y em plantas de batata cultivadas in vitro

    Luciana Cordeiro Nascimento

    2003-01-01

    Full Text Available Potato cultivars (Solanum tuberosum L. have shown degeneration or run out caused by viruses after several cycles of propagation using seed tubers from commercial fields. This work reports the occurrence of single and mixed infections of four potato viruses in Paraíba-Brazil and presents a method for Potato virus Y (PVY elimination, by using thermo-and chemotherapies. Plants of potato cv. Baraka were tested by direct antigen coating ELISA. Antisera against PVY, Potato virus X (PVX, Potato virus S (PVS, and Potato leafroll virus (PLRV were used. Materials with positive reaction to PVY were treated for virus elimination. Single node cuttings (1.0 cm length were excised and inoculated in Murashige & Skoog (MS medium, supplemented with 1.0 mg L-1 of kinetin, 0.001 mg L-1 of naphthalene acetic acid (NAA and 0.1 mg L-1 of gibberellic acid (GA3. The thermotherapy at approximately 37ºC, during 30 and 40 days, resulted in 20.0 and 37.5% PVY elimination, respectively. Chemotherapy was undertaken with Ribavirin (RBV, 5-Azacytidine (AZA, and 3-Deazauridine (DZD. The RBV showed the highest rate of virus eradication, with 55.5% virus-free plants. Simultaneous thermo and chemotherapy had higher efficiency for the elimination of PVY, reaching rates of healthy plants of 83.3% with RBV, 70.0% with AZA, and 50.0% with DZD.Cultivares de batata (Solanum tuberosum L. têm mostrado degenerescência causada por vírus após ciclos sucessivos do uso de tubérculos de campos comerciais como material propagativo. Este trabalho verifica a ocorrência de infecção simples e mista de quatro vírus da batata na Paraíba e apresenta adequação da técnica de cultivo in vitro para obtenção de material livre de Potato virus Y (PVY, incluindo uso de microestacas, termo e quimioterapia. Plantas de batata do cv. Baraka foram submetidas à indexação sorológica pelo teste "direct antigen coating" ELISA. Utilizaram-se antissoros contra o PVY, Potato virus X (PVX, Potato virus

  18. Biorregulador na composição química e na produtividade de grãos de soja Plant growth regulator in the chemical composition and yield of soybeans

    Leandro Paiola Albrecht

    2012-12-01

    was used, consisting of three plant growth regulators in the following concentrations: 0.005% indolebutyric acid IBA - IBA (auxin analogue, 0.009% kinetin (cytokinin and 0.005% gibberellic acid - GA3 (gibberellin. The use of plant growth regulator had an influence on the increase in productivity; oil and protein levels were altered by the action of the regulator, which tended to favour the protein content.

  19. PRODUÇÃO IN VITRO DE MUDAS DE Cattleya walkeriana E Cyrtopodium palmifrons A PARTIR DE SEMENTES IN VITRO PRODUCTION OF SEEDLINGS OF Cattleya walkeriana AND Cytopodium palmifrons FROM SEEDS

    Iraídes Fernandes Carneiro

    2007-09-01

    Full Text Available

    Sementes de Cattleya walkeriana e de Cyrtopodium palmifrons (Orchidaceae foram transferidas assepticamente para meios de cultura Knudson, MS (Murashige & Skoog e MS suplementado com IAA (1, 15 e 30 mg L-1 e cinetina (0,25, 5 e 10 mg L-1, para germinação. Elas foram incubadas à temperatu ra de 23 +/- 2°C, sob luz fluorescente por um período de 30 a 45 dias. As plântulas obtidas foram repicadas para meio Knudson, permanecendo em incubação por quatro meses e, em seguida, transferidas para meio fresco, até atingirem tamanho para aclimatação. O cultivo in vitro de ambas as espécies foi considerado viável, havendo comportamento diferente entre as espécies, em relação à produção de protocormóides, níveis de oxidação e crescimento in vitro. Os resultados mostraram que um meio me nos concentrado como Knudson pode ser utilizado sem adição de reguladores de crescimento para produção de mudas tanto para Cattleya walkeriana quanto para Cyrtopodium palmifrons.

    PALAVRAS-CHAVE: Orchidaceae; cultura de tecidos; orquídea.

    Seeds of Cattleya walkeriana and Cyrtopodium palmjfrons (Orchidaceae were aseptically transferred to culture medium Knudson, MS (Murashige& Skoog and MS supplemented with IAA (l, 15 and 30 mg L-1 and kinetine (0.25, 5 and 10 mg L-1 for germination. They were kept at room temperature (23 +/- 2°C under fluorescent light for 30 to 45 days. The seedlings were transferred to pots containing culture medium Knudson and incubated for four months at room temperature and transferred to fresh culture medium until the seedlings attained approximately 1.0 to 2.0 cm size for

  20. An improved type of ''Kohleria'' obtained through in vitro chemical mutagenesis

    Geier, T.

    1989-01-01

    Full text: Kohleria hybrids (Gesneriaceae) are used as indoor ornamentals. As in other pot plants, compact plant habit and low energy requirement are major breeding objectives. A trihybrid of the following composition was used in the experiments: (K. amabilis x K. bogotensis) x K. eriantha. This hybrid has attractive flowers but long internodes, large leaves and is late flowering under low light conditions. Adventitious buds were induced in high numbers by cultivating internode segments aseptically on agar medium supplemented with 1 mg/l kinetin and 0.5 mg/l IAA. For inducing mutations, internode explants from in-vitro grown shoots were soaked for 1 hour in a filter-sterilized aqueous solution of N-nitroso-N-methylurea (NMH; 500 mg/l) at 20 deg. C; the solution was unbuffered and immediately used after preparation. Two-node micro-cuttings of regenerated shoots were rooted ex vitro in a mixture of peat and sand (1:1 by volume) and grown into mature plants without losses. Whereas control plants regenerated in-vitro through adventitious buds showed as little variation as plants raised from conventional tip cuttings, conspicuous phenotypic changes were observed in plants originating from NMH-treated explants. In a first experiment, 21% of the plants differed from the controls in one or more characters. While the majority of these variants were inferior to the control, one designated as ''II-2-0'' showed early flowering under the natural low light conditions during winter. This character was retained after cloning. A second mutagenesis experiment was performed using aseptic stock cultures of this mutant ''II-20-0'', in order to obtain a more desirable plant habit. In this case, plants bearing phenotypic changes occurred about twice as frequently as in the first experiment, although the mutagenic treatment was the same. Among the selected variants there was one, designated ''II-2-32'', in which early flowering under low light conditions was coupled with significantly

  1. Rescate de embriones para la obtención de vitroplantas de vid (Vitis vinífera L.

    César Augusto Hernández Rendón

    2013-07-01

    grapevine Red Globe variety. The seeds were disinfected submerging them in 5 g/l of dicloroisocianuric acid (NaDCC for 15 min, and then in Benomyl® 2 g/l for 15 min, with an effectiveness of 92%. Different treatments were performed to obtain plants using seeds as explants, which were cultured on Murashige Skoog medium supplemented with different concentrations of indole acetic acid (AIA in combination with gibberellic acid (AG3 and kinetin (K without obtaining favorable answer for the germination. As alternative, immature seeds were extracted from grape fruits and placed on Murashige Skoog medium supplemented with 100 mg/l polyvinyl pyrrolidone (PVP, 0.35 mg/l of gibberellic acid and 1.75 mg/l of indol acetic acid for a month. Seeds were then opened and performed embryo rescue, planting them under dark conditions for eight days in the culture media Murashige Skoog 1 and 2 modified, finding plants formation by 40% a month after growing.

  2. Propagação in vitro de Limonium latifolium Kuntze (Plumbaginaceae In vitro propagation of Limonium latifolium Kuntze (Plümbaginaceae

    Claudimar Sidnei Fior

    2000-08-01

    Full Text Available Limonium latifoKum Kuntze é uma flor de corte cuja produção comercial de mudas é viabilizada através do cultivo de tecidos. Com o objetivo de aperfeiçoar o protocolo para a propagação clonal in vitro, fez-se uma sequência de estudos em que foram avaliados: viabilidade do uso de segmentos nadais do eixo da inflorescência imatura como explantes; tipos e concentrações de citocininas (cinetina-KIN e benzilaminopurina-BAP na regeneração; tipos e concentrações de auxinas (ácido naftalenoacético-ANA e ácido indolbutírico-AIB na fase de enraizamento in vitro; e técnicas de aclimatizaçao. Explantes oriundos de segmentos nadais da parte apical do eixo da inflorescência imatura são viáveis para a micropropagaçao de L. latifolium. Na fase de regeneração, os melhores resultados foram obtidos com o uso de meio de cultivo MS com 0,7mg de BAP/l por 35 dias. A fase de enraizamento m vitro foi feita com a manutenção dos explantes por 30 dias em meio MS acrescido de 1mg de AIB/l , garantindo ótimo índice de sobrevivência após a transferência para in vivo. A aclimatiwção foi feita sob cobertura plástica, em bancada de casa de vegetação, à temperatura ambiente e com irrigação por nebulização. Utilizaram-se, como recipientes, bandejas multicelulares de isopor, com 242 células e aproximadamente 10cm³ cada, preenchidas com casca de arroz carbonizada e esterilizada. As mudas receberam, quinzenalmente, adubação líquida com 0,5g/l de um adubo comercial (15:5:l5+micronutrientes. O processo in vitro dura 100 a 120 dias e um explante origina 15 a 30 mudas.Limonium latifolium Kuntze is a cut flower commercialy propagated in vitro. To develop and improve the micropropagation protocol, a sequence of assays was developed to evaluate performance of node explants; effect of cytokinins concentration (kinetin-KlN and 6-benzyl-aminopurin-BA on the regeneration rate; the presence of BA during the multiplication phase; concentration of

  3. Growth characteristics and dynamics of protein synthesis in callus cultures from Glycine wightii (Wight & Arn. Verdc. Características do crescimento e dinâmica da síntese de proteínas em cultura de calos de Glycine wightii (Wight & Arn. Verdc.

    André Luis Coelho da Silva

    2005-12-01

    Full Text Available Cotyledon explants were first cultured on MS medium supplemented with 4.52 M 2,4-D and 0.46 mM kinetin. The development of the calli was followed (0, 4, 8, 12, 16, 20, 24, 28 and 32 days after and the growth curve was determined, based in fresh and dry weight. The growth curve presented sigmoidal form with four distinct phases. The highest growth percentage was observed at the exponential phase and the lowest at the stationary phase. These results indicated that cotyledon callus subculture should be performed 20 days after inoculation. The calli obtained after a period of 28 days were freeze dried, macerated and submitted to extraction with buffers of different pH values (2.6; 4.0; 6.0; 8.0 and 10.0 and the proteins in the extracts were determined by Bradford method. The pH 8.0 buffer was the most efficient to extract the largest amount of protein. The amino acid analyses calli showed a high content of aspartic acid and low content of metionin. The dynamics of protein synthesis in calli was followed by SDS-PAGE electrophoresis.Explantes cotiledonários foram primeiramente cultivados em meio MS suplementado com 2,4-D 4,52 mM e cinetina 0,46 mM. O desenvolvimento dos calos foi acompanhado (0, 4, 8, 12, 16, 20, 24, 28 até 32 dias após o subcultivo e uma curva de crescimento foi determinada com base nos pesos fresco e seco dos calos. A curva de crescimento apresentou um padrão sigmoidal com quatro fases distintas. A maior porcentagem de crescimento foi observada na fase exponencial e a menor na fase estacionária. Estes resultados indicam que o subcultivo dos calos pode ser realizado 20 dias após a inoculação. Os calos obtidos após um período de 28 dias foram liofilizados, macerados e submetidos à extração de proteínas solúveis, utilizando tampões com diferentes valores de pH (2,6; 4,0; 6,0; 8,0 e 10,0 e o teor protéico dos extratos foi determinado pelo método de Bradford. O tampão pH 8,0 foi o mais eficiente na extração de prote

  4. Plant biostimulant associated with a resistance inducer in the production components of the common bean = Bioestimulante vegetal associado a indutor de resistência nos componentes da produção de feijoeiro

    João William Bossolani

    2017-10-01

    Full Text Available In the search for increased productivity in the common bean and in various other crops of economic interest, much research has been carried out using growth regulators, hormones and nutrients, especially in foliar application, with the aim of accelerating plant development, improving agronomic characteristics, and ensuring better plant health and performance in the field. The application of biostimulant plant regulators with the purpose of increasing crop productivity is a still not a common practice in agriculture. The aim of this study therefore, was to evaluate the effect of the application of plant biostimulants together with a resistance inducer on the production components of the common bean. Five doses of a biostimulant (indolebutyric acid, kinetin and gibberellic acid were used (0, 0.5, 1.0, 1.5 and 2.0 L ha-1, combined with four doses of a resistance inducer (0.5, 1.0, 2.0 and 3.0 L ha-1 in the common bean “IAC Formoso” under a no-tillage system. The experiment was carried out in Selvíria, in the State of Mato Grosso do Sul, during the 2013/14 agricultural year. The experimental design was of randomised blocks, in a 5x4 factorial scheme, with four replications. Production components and seed productivity were evaluated in the crop. The resistance inducer gave higher values for green matter (52.08 g plant-1 and dry matter (17.29 g plant-1 in the common bean at a dose of 2.04 L ha-1 and 2.15 L ha-1 respectively. The greatest productivity (3,756 kg ha-1 was obtained with a dose of 0.78 L ha-1 of biostimulant. The use of vegetable biostimulant can be a viable alternative for increasing productivity in the common bean. Em busca do aumento da produtividade do feijoeiro e de várias outras culturas de interesse econômico, foram realizadas inúmeras pesquisas utilizando reguladores de crescimento, hormônios e nutrientes, principalmente em aplicações foliares com a finalidade de acelerar o desenvolvimento da planta, melhorar suas

  5. USO DE REGULADOR DE CRESCIMENTO EM CULTIVARES DE FEIJÃO DE INVERNO PLANT GROWTH STIMULANT APPLICATION ON WINTER COMMON BEAN CULTIVARS

    Salatiér Buzetti

    2011-04-01

    contradictory. For that reason, the objective of this study was to evaluate the effects of plant growth stimulant application on components and grains yield of two winter common bean cultivars under Brazilian savannah conditions. The experiment was conducted during the 2007 fall-winter season, at the Unesp experimental farm, Ilha Solteira campus, in Selvíria, Mato Grosso do Sul State, Brazil. The experimental design was randomized blocks, in a strips scheme, with four replications. The treatments consisted of a combination of five plant growth stimulant doses (0 L ha-1; 0.5 L ha-1; 1.0 L ha-1; 1.5 L ha-1; and 2.0 L ha-1, composed of three vegetable hormones (kinetin, gibberellic acid, and indolbutyric acid, in two application periods: at the vegetative stage (V4 and at the reproductive stage (R5. Vegetative characteristics such as plant height, first pod insertion height, number of grains per pod, and weight of 100 grains were not affected by the product application. However, its application at the reproductive stage (R5 increased the number of grains per plant and grain yield of the Carioca Precoce and IAC Apuã bean cultivars. For that increase, 2 L ha-1 was the best plant growth stimulant dose.

    KEY-WORDS: Phaseolus vulgaris; yield components; application period.

  6. Study of Cytokinin and Auxin Hormones and Planting Pattern Effects on Yield and Yield Components of Grain Maize (Zea mays L. under Saline Conditions

    D Davani

    2016-07-01

    Full Text Available Introduction Maize (Zea mays L. which belongs to the Poaceae family is the third important cereal crop of the world after wheat and rice. Salinity is one of the major environmental factors limiting plant growth and productivity. Maize is sensitive to salinity. Planting method is a crucial factor for improving crop yield. Planting methods in saline and non-saline conditions are different. Kinetin is one of the cytokinins known to significantly improve the growth of crop plants grown under salinity. Indole acetic acid (IAA is also known to play a significant role in plant tolerance to salt stress. However, little information appears to be available on the relationship between salinity tolerance and auxin or cytokinins levels in plants. In this respect, the objective of this study was to study the effects of foliar applications of cytokinin and auxin hormones on yield and yield components of grain maize under different planting patterns in saline conditions. Materials and Methods The experiment was carried out at Bushehr Agricultural and Natural Resources Research Center, Dashtestan station with 29° 16´ E latitude and 51° 31´ N, longitude and 70 m above the see surface during the 2013 growing season. Dashtestan region is a warm-arid region with 250 mm precipitation per year. The field plowed by April 2013 and then prepared and sowed by August 2013. There were five rows with 75 cm distance. The experiment was conducted as a split-plot factorial design based on complete randomized blocks with three replications. Planting pattern (ridge planting, double rows of planting on a ridge in zigzag form and furrow planting as the main factor and time of cytokinin (0 as a control, V5- V6 stage and V8- V10 stage and auxin (0 as a control, silking stage, two weeks after silking stage foliar-applied was considered in a factorial. Cytokinin (Benzyl Adenine, Merck and Auxin (Indole-3-Butiric Acid, Merck were sprayed on the entire plant in the evening with

  7. Effect of Different Dormancy Breaking Treatments on Seed Germination of Salsifis (Teragopogon collinus and Flixweed ( Descurainia Sophia as Two Weed Species

    marzie mazhari

    2017-01-01

    not the seeds are placed in layers. Chilling plays an important role in providing the stimulus required to overcome dormancy, increase germination, and produce normal seedlings for Prunuspersica. Exogenous growth regulator treatments – gibberellins (usually gibberellic acid GA3 and GA4+7 and cytokinins (usually kinetin, benzyladenine – have been shown to break dormancy in many seed species. The objective of the present study was to investigate effective methods in breaking the seed dormancy of Teragopogon collinus and Descurarina sophia and evaluate their response to different dormancy-breaking treatments, germination under different constant and alternating temperature, and using chemical, with most appropriate dormancy-breaking procedure for both species. Materials and Methods: In order to evaluate the effect of different treatments on dormancy breaking and seed germination of Teragopogon collinus and Descurarina sophia an experiment was carried out as factorial based on a Randomized Complete Block Design (RCBD with three replications at Shahrekord University in 2012. Treatments including different levels of gibberellic acid (0, 100, 250 and 500 mg at two soaking time levels of 2 and 24 hour (along with scarification pre-treatment, concentrations of potassium nitrate (50, 100 and 150 mM in 10 and 30 minutes (along with scarification pre-treatment, treatment of rotational temperature (5-15 and 10-20 °C, in full light, dark and rotational light periods (8 hr light and 16 hr dark and mechanical treatments of dormancy breaking (application of boiling water. Results and Discussion: Teragopogon collinus and Descurainia sophia had highest seed dormancy breaking under rotational temperature and light and application of gibberellic acid and potassium nitrate (along with scarification pre-treatment had no effect on Teragopogon collinus and Descurarina sophia dormancy. According to the Teragopogon collinus and Descurainia sophia seeds dormancy breaking in mechanical

  8. Premilinary studies for optimiziing a protocol for obtaining embryogenic calluses in two rubber (Hevea brasiliensis Mull. Arg clones from different geographical origins Estudios preliminares en la estandarización de un protocolo para la obtención de callos embriogénicos en dos clones de caucho (Hevea brasiliensis Müll. Arg. de diferentes orígenes geográficos

    Medina S. Marisol

    2006-07-01

    Full Text Available The influence of growth regulators on obtaining friable rubber (Hevea brasiliensis Müll. Arg. calluses with no plant regeneration as investigated. Two clones having different geographical origin were used in all trails carried out in this study: FX 3864 (South-American and PB 254 (Asian. Young leaves and eight- to ten-week-old seed integument from both clones were used as explants in several experiments; they were initially cultured in MH medium (Carron, et ál., 1989, modified MH medium (Montoro, et ál., 1993, 2000 and modified MS medium (Carron, et at,. 1992, no positive response being obtained by days 25 or 50. However, other trials were carried out with the integument in modified MS medium (1962, 0.67 mg/L BAP and 0.66 mg/L 2-4 D being added as medium for initiating embryogenesis, the formation of white, friable calluses being observed by day 25 in the two selected clones. These calluses were sub-cultured in MS supplemented with 0.35 mg/L BAP and 0.2 mg/L 2-4 D as callogenesis expression medium, embryogenic expression being observed in both clones by day 50. Equally friable white calluses were obtained from young leaves in the two clones in MS medium supplemented with 1.0 mg/L BAP, 1.0 mg/L ANA but without IBA and kinetin by day 25. Calluses sub-cultured in the same medium supplemented with 0.5 mg/L BAP and 0.5 mg/L ANA began to show increased friability after 50 days. culture. This work is a partial report of a macro-project for optimising a protocol for rubber (Hevea brasiliensis  multiplication by somatic embryogenesis.Se investigó la influencia de los reguladores de crecimiento para la obtención de callos friables en caucho (Hevea brasiliensis Müll. Arg. sin llegar a obtenerse regeneración de plántulas. En todos los ensayos realizados en este estudio, se utilizaron dos clones de diferentes orígenes geográficos: el FX 3864 (suramericano y el PB 254 (asiático. Para los diferentes tratamientos se utilizaron como explantes hojas j

  9. DEVELOPMENT OF GENOMIC AND GENETIC TOOLS FOR FOXTAIL MILLET, AND USE OF THESE TOOLS IN THE IMPROVEMENT OF BIOMASS PRODUCTION FOR BIOENERGY CROPS

    Chen, Xinlu; Zale, Janice; Chen, Feng

    2013-01-22

    Foxtail millet (Setaria italica L.) is a warm-season, C4 annual crop commonly grown for grain and forage worldwide. It has a relatively short generation time, yet produces hundreds of seeds per inflorescence. The crop is inbred and it has a small-size genome (~500 Mb). These features make foxtail millet an attractive grass model, especially for bioenergy crops. While a number of genomic tools have been established for foxtail millet, including a fully sequenced genome and molecular markers, the objectives of this project were to develop a tissue culture system, determine the best explant(s) for tissue culture, optimize transient gene expression, and establish a stable transformation system for foxtail millet cultivar Yugu1. In optimizing a tissue culture medium for the induction of calli and somatic embryos from immature inflorescences and mature seed explants, Murashige and Skoog medium containing 2.5 mg l-1 2,4-dichlorophenoxyacetic acid and 0.6 mg l-1 6- benzylaminopurine was determined to be optimal for callus induction of foxtail millet. The efficiency of callus induction from explants of immature inflorescences was significantly higher at 76% compared to that of callus induction from mature seed explants at 68%. The calli induced from this medium were regenerated into plants at high frequency (~100%) using 0.2 mg l-1 kinetin in the regeneration media. For performing transient gene expression, immature embryos were first isolated from inflorescences. Transient expression of the GUS reporter gene in immature embryos was significantly increased after sonication, a vacuum treatment, centrifugation and the addition of L-cysteine and dithiothreitol, which led to the efficiency of transient expression at levels greater than 70% after Agrobacterium inoculation. Inoculation with Agrobacterium was also tested with germinated seeds. The radicals of germinated seeds were pierced with needles and dipped into Agrobacterium solution. This method achieved a 10% transient

  10. Propagación vegetativa in vitro de Puya santossi Puya santossi in vitro propagation

    Pedroza–Manrique Jaime Alonso

    2008-07-01

    ías de extinción; reguladores de crecimiento.Puya santossi (L is an endemic threatened shrub from the high Andean region. Conservation procedures must thus be established for this important germplasm species’ conservation. This study evaluated the in vitro adaptation of this species with Benlate and nine sodium hypochlorite treatments and the effect of five growth regulators on the morphogenetic development of shoots, meristems and leaves. The base medium used was MS enriched with the following growth regulators: indol butyric acid (IBA and benzylaminopurine (BAP (0.0, 0.5, 1.0 and 1.5 mg/L in meristem culture; α-naphthaleneacetic acid (NAA (0.0, 1.0, 2.0 and 3.0 mg/L in the rooting of buds regenerated in vitro; 2, 4-dichlorophenoxy acetic acid and Kinetin (0.0, 1.0, 3.0 and 5.0 mg/l in leaves. It was observed that treatment 5 (1% NaOCl, 10 min was the most successful in contamination control. The meristem was the most appropriate explant for Puya santossi micropropagation, allowing mass proliferation of adventitious buds when they were grown on MS media enriched with 1.5 mg/L BAP. Buds from the meristem culture were rooted with 3.0 mg/L NAA. This protocol represents a conservation model for endangered species and led to propagating Puya santossi. This investigation revealed that morphogenetic leaf explants were inhibited by the presence of a group of chlorophycea algae which were found to be symbiotically associated with P. santossi. Key words: Puya santossi; micropropagation; endangered plant; growth regulators