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Showing papers on "Kinetin published in 2013"


Journal ArticleDOI
TL;DR: In ARs formed in planta and TCLs, the QC is established in a similar way, and auxin transport and biosynthesis are involved through cytokinin tuning.

137 citations


Journal ArticleDOI
TL;DR: Investigation of anti-inflammatory effect of the alcoholic extracts of N. sativa seeds and its callus on mix glial cells of rat with regard to their thymoquinone (TQ) content found decrease in the TQ content of the callus was accompanied with an increase in its phenolic content and antioxidant ability.
Abstract: Anti-inflammatory effect of the alcoholic extracts of N. sativa seeds and its callus on mix glial cells of rat with regard to their thymoquinone (TQ) content was investigated. Callus induction was achieved for explants of young leaf, stem, petiole, and root of N. sativa on solid Murashige and Skoog (MS) medium containing 2,4-D (1 mg/l) and kinetin (2.15 mg/l). TQ content of the alcoholic extracts was measured by HPLC. Total phenols were determined using Folin–Ciocalteu method and antioxidant power was estimated using FRAP tests. The mix glial cells, inflamed by lipopolysaccharide, were subjected to anti-inflammatory studies in the presence of various amounts of TQ and the alcoholic extracts. Viability of the cells and nitric oxide production were measured by MTT and Griess reagent, respectively. The leaf callus obtained the highest growth rate (115.4 mg/day) on MS medium containing 2,4-D (0.22 mg/l) and kinetin (2.15 mg/l). Analyses confirmed that TQ content of the callus of leaf was 12 times higher than that measured in the seeds extract. However, it decreased as the calli aged. Decrease in the TQ content of the callus was accompanied with an increase in its phenolic content and antioxidant ability. Studies on the inflamed rat mix glial cells revealed significant reduction in the nitric oxide production in the presence of 0.2 to 1.6 mg/ml of callus extract and 1.25 to 20 μl/ml of the seed extracts. However, the extent of the effects is modified assumingly due to the presence of the other existing substances in the extracts.

74 citations


Journal ArticleDOI
TL;DR: Investigation of the effects of salicylic acid and methyl jasmonate on anthocyanin induction, biomass accumulation, and color value (CV) indices for both pigment content (PC) and pigment production (PP) in callus cultures of Rosa hybrida cv.
Abstract: This study was undertaken to investigate the effects of salicylic acid (SA) and methyl jasmonate (MeJA) on anthocyanin induction, biomass accumulation, and color value (CV) indices for both pigment content (PC) and pigment production (PP) in callus cultures of Rosa hybrida cv. Pusa Ajay. A concentration-dependent response was exhibited by cultures on SA and MeJA at different concentrations individually or in combinations to Euphorbia millii medium supplemented with 204.5 mM sucrose, 2.45 μM indole butyric acid and 2.33 μM kinetin. There was positive influence on both callus biomass and anthocyanin accumulation. Treatment with 0.5 μM MeJA was most effective in inducing anthocyanin biosynthesis in callus cultures. Anthocyanin accumulation in callus cultures was enhanced with the addition of SA and MeJA, but these did not differ significantly from control for the number of days required for pigment initiation and for color intensification. Moreover, the addition of 0.5 μM MeJA alone resulted in a higher frequency of color response (97.25 %), PC (3.48 ± 0.07 CV g−1 FW), and PP (1.56 ± 0.03 CV test tube−1) over control. In contrast, the presence of higher levels of SA (400 μM) and MeJA (5.0 μM) reduced frequency of color response, as well as levels of PC and PP. MeJA did not increase biomass accumulation but promoted frequency of color response, PC and PP. Hence, it was suggested that 0.5 μM MeJA promoted anthocyanin production in rose callus cultures. Significant correlation was found between frequency of response and each of the PC (r = 0.988) and PP (r = 0.990). Furthermore, PC and PP were also highly correlated (r = 0.998).

68 citations


Journal ArticleDOI
TL;DR: A rapid and efficient micropropagation protocol for M. peregrina has been established and no polymorphism was detected indicating the genetic integrity of in vitro propagated plants.
Abstract: Moringa peregrinais an endangered species of Moringaceae.M. peregrinais a multipurpose tree with a wide variety of potential uses including its medicinal activity. In our study, a rapid and efficient micropropagation protocol for M. peregrina has been established. In vitro germinated seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different levels of either 6-benzyladenine (BA) or kinetin (Kin). The maximum shoot proliferation of 6.5 shoots per explant with 100 % shoot proliferation rate was observed on MS medium supplemented with 1.0 mg/l BA. On the other hand, MS medium supplemented with 1 mg/l indole-3-butyric acid (IBA) resulted in the maximum number of roots. Micropropagated plants were successfully acclimatized. Genetic stability of micropropagated plants was assessed using Inter-Simple Sequence Repeat (ISSR). The amplification products were monomorphic in all in vitro grown plants. No polymorphism was detected indicating the genetic integrity of in vitro propagated plants. This micropropagation protocol could be useful for raising genetically uniform plants for plant propagation and commercial cultivation.

54 citations


Journal ArticleDOI
TL;DR: The effect of culture medium nutrients on growth and alkaloid production by plant cell cultures of Nothapodytes nimmoniana (J. Grah.) Mabberley (Icacinaceae) was studied with a view to increasing the production of the alkaloids camptothecin, a key therapeutic drug used for its anticancer properties.
Abstract: The effect of culture medium nutrients on growth and alkaloid production by plant cell cultures of Nothapodytes nimmoniana (J. Grah.) Mabberley (Icacinaceae) was studied with a view to increasing the production of the alkaloid camptothecin, a key therapeutic drug used for its anticancer properties. Amongst the various sugars tested with Murashige and Skoog (MS) medium, such as glucose, fructose, maltose, and sucrose, maximum accumulation of camptothecin was observed with sucrose. High nitrate in the media supports the biomass, while high ammonium enhances the camptothecin content. Selective feeding of 60 mM total nitrogen with a NH4+/NO3− balance of 5/1 on day 15 of the culture cycle results in a 2.4-fold enhancement in the camptothecin content over the control culture (28.5 μg/g DW). Furthermore, the sucrose feeding strategy greatly stimulated cell biomass and camptothecin production. A modified MS medium was developed in the present study, which contained 0.5 mM phosphate, a nitrogen source feeding ratio of 50/10 mM NH4+/NO3− and 3 % sucrose with additional 2 % sucrose feeding (added on day 12 of the cell culture cycle) with 10.74 μM naphthaleneacetic acid and 0.93 μM kinetin. Finally, the selective medium has 1.7- and 2.3-fold higher intracellular and extracellular camptothecin content over the control culture (29.2 and 8.2 μg/g DW), respectively.

54 citations


Journal ArticleDOI
TL;DR: A novel large-scale micropropagation pathway for date palm (Phoenix dactylifera L.) based on organogenesis is developed and the survival rate after 2 months was related to the medium used during the elongation phase; >90 % of shoots that were cultured on PGR-free media survived, while there was a poor survival rate of shoot that had been cultured on media containing PGRs.
Abstract: We developed a novel large-scale micropropagation pathway for date palm (Phoenix dactylifera L.) based on organogenesis. We obtained organogenic stems from shoot tip explants of the Moroccan date palm cultivar Najda, and investigated shoot proliferation from these organogenic stems in vitro on various media; Beauchesne medium (BM) and Murashige and Skoog medium (MS) at full-strength, half-strength, and one-third-strength, containing various concentrations (0, 0.25, 0.5, and 1 mg/L) of 2-naphthoxyacetic acid (NOAA) and kinetin. The optimal medium during the multiplication phase was half-strength Murashige and Skoog medium (MS/2) supplemented with 0.5 mg/L NOAA and 0.5 mg/L kinetin (23.5 morphologically superior shoots per explant, with low vitrification rates). For the shoot elongation phase, shoots were transferred to the same proliferation medium, or to MS or MS/2 media without plant growth regulators (PGRs). Shoots elongated rapidly and showed a high rate of root formation on media supplemented with PGRs. For example, on MS/2 medium containing 1 mg/L NOAA and 1 mg/L kinetin, the average shoot length was 15.1 cm, the average number of roots per shoot was 6.2, and their average length was 3.4 cm. On PGR-free media, shoots were shorter with wider and greener leaves, and had fewer roots. The plantlets were transferred to a greenhouse for acclimation. The survival rate after 2 months was related to the medium used during the elongation phase; >90 % of shoots that were cultured on PGR-free media survived, while there was a poor survival rate of shoots that had been cultured on media containing PGRs.

47 citations


Journal ArticleDOI
TL;DR: The results show that in vitro propagated and hardened plants of S. rebaudiana are morphologically as well as functionally comparable to each other and to their mother plant.
Abstract: An efficient high frequency plant regeneration protocol through direct organogenesis was developed for Sevia rebaudiana Bert. Nodal segments containing axillary buds were used as an explant and inoculated on Murashige and Skoog’s (MS) medium containing 3% (w/v) sucrose, 0.8% (w/v) agar supplemented with various concentrations of benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) ranging from 1.00 to 9.00 μM. Maximum multiple shoots (96%) were obtained in MS medium supplemented with 1.0 μM TDZ with an average of 60 shoots per culture, having an average shoot length of 6.0 cm. The best in vitro root induction (89%) was achieved on half strength MS medium without any growth regulator with an average of 24 roots per culture and root length of7 cm. The rooted plantlets were successfully established in soil and grown to maturity at the survival rate of 95% in the indoor grow room. High-performance liquid chromatography was used to assess the stability in chemical profile and quantification of stevioside and rebaudioside A content of in vitro propagated S. rebaudiana plants and compared with their mother plant at the peak vegetative stage. Our results show no significant differences (p in vitro propagated plants. Furthermore, fully developed in vitro propagated S. rebaudiana plants were also compared with mother plant for their gas and water vapour exchange characteristics and leaf anatomy. The results show that in vitro propagated and hardened plants of S. rebaudiana are morphologically as well as functionally comparable to each other and to their mother plant.

46 citations


Journal ArticleDOI
TL;DR: A significantly high IC50 value of 170.13 μg/ml was attained in normal cell line VERO indicating that its natural counterpart is an ideal candidate for treatment of brain cancer without causing negative effects to normal healthy cells.
Abstract: Black carrots contain anthocyanins possessing enhanced physiological activities. Explants of young black carrot shoots were cultured in Murashige and Skoog (MS) medium for callus initiation and were transferred to new MS medium supplemented with four different combinations of 2,4-dichlorophenoxyacetic acid and kinetin. Subsequently, the lyophilized calli and black carrot harvested from fields were subjected to ultrasound extraction with ethanol at a ratio of 1:15 (w:v). Obtained extracts were applied to various human cancer cell lines including MCF-7 SK-BR-3 and MDA-MB-231 (human breast adenocarcinomas), HT-29 (human colon adenocarcinoma), PC-3 (human prostate adenocarcinoma), Neuro 2A (Musmusculus neuroblastoma) cancer cell lines and VERO (African green monkey kidney) normal cell line by MTT assay. The highest cytotoxic activity was achieved against Neuro-2A cell lines exhibiting viability of 38-46% at 6.25 μg/ml concentration for all calli and natural extracts. However, a significantly high IC50 value of 170.13 μg/ml was attained in normal cell line VERO indicating that its natural counterpart is an ideal candidate for treatment of brain cancer without causing negative effects to normal healthy cells.

44 citations


Journal ArticleDOI
Anwar A. Khan1
TL;DR: ABA abscisic acid. as discussed by the authors is an acid that is a mixture of Abscisicsic acid and Cotylenin (COTYLENIN) and it has been shown to have light osmotic treatment.
Abstract: ABA abscisic acid. CN cotylenin E. CP cordycepin. DOT dark osmotic treatment. E ethephon. FC fusicoccin. GA gibberellic acid. K kinetin. LOT light osmotic treatment. PEG polyethylene glycol-6000. P...

38 citations


Journal ArticleDOI
TL;DR: To induce somatic embryogenesis, shoot organogenesis and plant regeneration for Camellia nitidissima, three types of callus were induced from immature cotyledons on improved woody plant medium (WPM) with different plant growth regulators (PGRs).

36 citations


Journal ArticleDOI
TL;DR: This protocol is an efficient means for the large-scale propagation of P. hangianum and about 5000 plantlets were successfully produced within 3 years from plant regeneration, showing no obvious phenotypic variation.

Journal ArticleDOI
TL;DR: Histological studies revealed the origin of shoot tip in the callus during regeneration of Withania somnifera (L.) Dunal, an important medicinal plant being the source of extremely important compounds like withanolides and withaferin.
Abstract: Withania somnifera (L.) Dunal, is an important medicinal plant being the source of extremely important compounds like withanolides and withaferin. Influence of different plant growth regulators (PGRs) were evaluated for induction of callus, callus mediated regeneration and production of secondary metabolites in them. Explants for callusing were collected from plants grown in vitro and maximum callusing (98 %) was obtained on MS medium supplemented with a combination of 2,4-dichlorophenoxy acetic acid (2,4-D) (0.5 mg l-1) and kinetin (KN) (0.2 mg l−1). Among different types of calli, best shoot regeneration was observed on green, compact calli produced on MS medium with a combination of 6-benzylamino purine (BAP) and indole butyric acid (IBA). MS medium supplemented with BAP (2 mg l−1) showed highest frequency (98 %) of shoot bud regeneration. The micro-shoots were efficiently rooted on MS media supplemented with 0.5 mg l−1 IBA. Rooted plants were transferred to soil-vermi-compost (1:3; w/w) medium in greenhouse for acclimatization. Presence of withanolide A and withaferin A in calli was validated through high performance thin layer chromatography (HPTLC). It was interesting to observe that the PGRs showed significant influence on the secondary metabolites production in callus and 2,4-D having the least effect. Histological studies revealed the origin of shoot tip in the callus during regeneration.

Journal ArticleDOI
TL;DR: Immature cotyledons collected from four genotypes of chickpea were cultured adaxially on Murashige and Skoog medium supplemented with 6-benzyladenine, thidiazuron, kinetin, zeatin and dimethylallylaminopurine for dedifferentiation and regeneration of adventitious shoots and the initiation of meristematic activity in the sub-epidermal region during the onset of morphogenesis was revealed.
Abstract: Immature cotyledons collected at different time intervals from four genotypes of chickpea (C 235, BG 256, P 362 and P 372) were cultured adaxially on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine, thidiazuron, kinetin, zeatin and dimethylallylaminopurine (2-iP), either alone or in combination with indole-3-acetic acid (IAA) or α-napthoxyacetic acid (α-NOA) for dedifferentiation and regeneration of adventitious shoots. Morphogenesis was achieved with explants cultured adaxially on MS medium with 13.68 μM zeatin, 24.6 μM 2-iP, 0.29 μM IAA and 0.27 μM α-NOA. Explants prepared from pods of 21 days after pollination, responded favourably to plant growth regulator treatment in shoot differentiation. Histological studies of the regenerating explants, revealed the initiation of meristematic activity in the sub-epidermal region during the onset of morphogenesis, which can be correlated with elevated activity of cytokinin oxidase-dehydrogenase, for cytokinin metabolism. The regenerated shoots were efficiently rooted in MS medium supplemented with 2.46 μM indole-3-butyric acid and acclimatized under culture room and glasshouse conditions for normal plant development leading to 76–80 % survival of the rooted plantlets. The immature cotyledon explants were used for Agrobacterium-mediated transformation with critical manipulation of cultural conditions like age of explant, O.D. of Agrobacterium suspension, concentration of acetosyringone, duration of sonication and co-cultivation for successful genetic transformation and expression of the reporter gene uidA (GUS). Integration of transgene was confirmed by molecular analysis. Transformation frequency up to 2.08 % was achieved in chickpea, suggesting the feasibility of using immature cotyledon explants for Agrobacterium-mediated transformation.

01 Jan 2013
TL;DR: Kinetin was found to be the most affective in increasing growth parameter and nitrogen fixation of chickpea and both the efficiency and the longevity of the nodules seem to be favorably affected by kinetin application.
Abstract: The study was conducted to compare the effects of three plant growth regulators and Rhizobium leguminosarum on the growth, yield parameters and N2 fixation of chickpea under natural condition. The plant growth regulators viz., kinetin, indole 3-acetic acid (IAA) and abscisic acid (ABA) were applied at concentration of 10 -5 M as seed soaking and 10 -6 M as foliar spray, alone and in combinations with Rhizobium inoculum (strain TAL 1148 and TAL 620). Kinetin was found to be the most affective in increasing growth parameter (viz. root/shoot biomass, grain yield) and nitrogen fixation (viz., specific nitrogenase activity of nodules, and total N – fixed plant -1 ) of chickpea. The IAA seed soaking was least effective. The ABA seed soaking as well as foliar spray treatments significantly decreased nodules weight, nitrogenase activity of nodules, specific nitrogenase activity of nodules and total N – fixed plant -1 . The kinetin seed soaking (10 -5 M) and ABA foliar spray (10 -6 M) were more effective in increasing the grain weight as compared to control. Application of Rhizobium inoculum, generally increased growth yields components and nitrogen fixation. The TAL 1148 strain was more effective than TAL 620. The pattern of response to hormone and Rhizobium inoculum was consistent in the three consecutive years. It would suggest that that both the efficiency and the longevity of the nodules seem to be favorably affected by kinetin application.

Journal ArticleDOI
TL;DR: This study provides the first report on C. attenuata shoot organogenesis and plant regeneration and indicated that all plantlets were diploid (2n = 84).
Abstract: Curcuma attenuata is a highly valued ornamental. This study provides the first report on C. attenuata shoot organogenesis and plant regeneration. Immature anthers derived from 5 to 7 cm long inflorescences were isolated and cultured on different variations of Murashige and Skoog (MS) media to induce callus and then shoot organogenesis. When the 2-mm long anthers in which microspores were at the uninucleate developmental stage were cultured in the dark on MS medium containing 13.6 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin (KT) for 15 days and then transferred to 40 μmol m−2 s−1 fluorescent light for 30 days, the percentage callus induction reached 33.3 %. After callus was transferred to various differentiation media and cultured in the light, 33.1 % of all callus cultures could differentiate into adventitious shoots on MS medium supplemented with 22.0 μM 6-benzyladenine (BA), 0.53 μM α-naphthaleneacetic acid (NAA) and 1.4 μM thidiazuron (TDZ) after culturing for 60 days. Over 95 % of plantlets survived after transplanting plantlets into trays with a mixture of sand and perlite (2: 1) for 20 days. Chromosome number, determined from the root tips of young plantlets, indicated that all plantlets were diploid (2n = 84).

Journal ArticleDOI
TL;DR: MS medium was observed to be the more effective in promoting the proliferation of somatic embryos than half-strength Murashige and Skoog (1/2MS), and timentin was also more efficient in induction of secondary embryogenesis than sucrose.
Abstract: Somatic embryogenesis was induced from in vivo grown leaf explants of Chrysanthemum cv. Euro incubated on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid and 2.0 mg/L Kinetin, yielding the highest mean number of embryos (42 ± 5.97) per explant after 5 weeks of culture. We evaluated the effects of basal medium, various concentrations of sucrose, and timentin on the proliferation of secondary somatic embryos. MS medium was observed to be the more effective in promoting the proliferation of somatic embryos than half-strength Murashige and Skoog (1/2MS). In addition, timentin was also more efficient in induction of secondary embryogenesis than sucrose. Whole plantlets were obtained by culturing of secondary embryos on hormone-free MS medium and successfully acclimated in the green house.

Journal ArticleDOI
TL;DR: The random amplified polymorphic DNA analysis of in vitro and in vivo plants showed four clusters of similarity and the largest number of regenerated plants was obtained from the embryos cultivated on media enriched with meta‐topolin and benzyladenine.
Abstract: Cytokinins are growth regulators that stimulate cell division and control morphogenesis in plants, however their role in regulating secondary metabolism is not well studied. The influence of various cytokinins (benzyladenine, zeatin, kinetin, meta-topolin, thidiazuron) and culture systems (solid and temporary immersion RITA (R) system) on the quality Leucojum aestivum plant regenerated from somatic embryos was investigated. The largest number of regenerated plants (181.6 and 168.8) was obtained from the embryos cultivated on media enriched with meta-topolin and benzyladenine. Thidiazuron and meta-topolin led to the highest number of normally developed plants (94.8 and 90.6). The random amplified polymorphic DNA analysis of in vitro and in vivo plants showed four clusters of similarity. The highest biomass (growth index: 2.49) was obtained with the temporary immersion RITA (R) system. Alkaloid extracts were analyzed by LC-MS, leading to the quantification of galanthamine and lycorine both in plant materials and in liquid media. The highest contents of galanthamine (0.05% dry weight) were observed in plants cultivated in the presence of thidiazuron in bioreactor system. Galanthamine was accumulated (highest content 0.05% dry weight) in plants cultivated in the presence of thidiazuron in bioreactor system whereas lycorine was synthetized mainly in plants cultivated on solid media.

Journal ArticleDOI
TL;DR: 2iP is the optimal plant growth regulator for Sophora multiplication and shows the best response for shoot multiplication among the four investigated cytokinins.
Abstract: Objective To determine the effects of different cytokinins at various concentrations on in vitro shoot multiplication of an important medicinal plant.

Journal Article
TL;DR: Of the different auxins (2, 4-D, NAA) and cytokinins (Kinetin, BAP) and their combinations studied, a ratio of 1:4 for 2,4-D and NAA and 1:3:1 ratio of Kinetin: BAP: NAA ratio proved to be optimal for callus induction and green plant regeneration respectively.
Abstract: Application of anther culture techniques for improvement of indica rices is a formidable task as they are known to be recalcitrant to culture unlike japonica rices. Since anther culture can effectively address the major problems associated with the adoption of hybrid rice in India, an effort was made to assess the influence of cold pretreatment and phyto hormones on the anther culture response of Rajalaxmi (CRHR 5) and Ajay (CRHR 7), two elite and popular indica rice hybrids. Cold pretreatment for 7-9 days at 10 o C was found to have a positive influence on the callus induction frequency irrespective of the media employed and prolonged treatment over the optimum proved to be inhibitory. Of the different auxins (2, 4-D, NAA) and cytokinins (Kinetin, BAP) and their combinations studied, a ratio of 1:4 for 2,4-D and NAA and 1:3:1 ratio of Kinetin: BAP: NAA ratio proved to be optimal for callus induction and green plant regeneration respectively. This information can be of use in the development of doubled haploids with superior yield and good grain quality from the elite hybrid cultivars.

Journal Article
TL;DR: Results revealed that IAA and/or Kin treatments had positive effects on growth criteria, photosynthetic pigments, total carbohydrate, polysaccharide, free amino acid, proline and total phenolic contents in the two faba bean cultivars.
Abstract: Two field experiments were conducted at the Research and Production Station, National Research Centre, Nubaria Province, Behaira Governorate, Egypt during two successive winter seasons 2011/2012 and 2012/2013 to study the effect of indole acetic acid (IAA) at 0, 50 and 75 mg/l and kinetin (Kin) at 0, 50, 75 and 100 mg/l individually or in combination on two faba bean cultivars (Nubaria 1 and Giza 843). Results revealed that IAA and/or Kin treatments had positive effects on growth criteria (plant height, leaves number, fresh and dry weight per plant), photosynthetic pigments (chlorophyll a, b and carotenoids), total carbohydrate, polysaccharide, free amino acid, proline and total phenolic contents in the two cultivars. Furthermore, endogenous hormones (IAA, GA and zeatin) were increased concomitantly with decrease in ABA content 3 particularly at treatment of 50 mg/l IAA + 75 mg/l kinetin in the two cultivars. Seed yield and yield components (number of pods/plant, pods yield /plant, 100 seeds weight and biological yield/plant) as well as total carbohydrates and proteins in the yielded seeds were significantly improved at all treatments in the two cultivars. Meanwhile, vicine contents were significantly decreased at all treatments. It is worthy to mention that, performance of Nubaria 1 cultivar was more pronounced than that of Giza 843 cultivar when grown under newly reclaimed sandy soil conditions i.e. Under control treatment, seed yield/faddan (one faddan= 0.42 ha) of Nubaria 1 cultivar was 47.17% over that of Giza 843 cultivar. Meanwhile, response of Giza 843 cultivar to all applied treatments was more effective than Nubaria 1 cultivar.

Journal ArticleDOI
TL;DR: No variation was reported among the in vitro raised progeny and the mother plant in the banding profiles generated by the total of fifteen Random Amplified polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers, which confirmed that these plants were genetically similar and can be used as elite plants.
Abstract:  Abstract—An efficient and reproducible protocol has been established through the technique of forced axillary branching for the propagation of an important edible bamboo species namely Bambusa bambos. High frequency multiple shoot induction was achieved from nodal segments collected from elite genotype on Murashige and Skoog’s (MS) medium supplemented with 4.4 µM Benzylaminopurine (BAP) and 1.16 µM Kinetin (Kn). The size of explant and season greatly influenced the frequency of bud break. Rooting posed a major problem to be worked out in this particular species. Best rooting response was observed on 9.80 µM of Indole- 3 Butyric acid (IBA) with 60 ± 14.1 % rooting. In vitro raised plants were successfully acclimatized and established in the field conditions where they exhibited normal growth. In a bid to ascertain genetic fidelity, DNA was extracted by CTAB method and samples were analysed in 1.8% agarose gel electrophoresis. In the present study no variation was reported among the in vitro raised progeny and the mother plant in the banding profiles generated by the total of fifteen Random Amplified polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers. Hence, molecular analysis confirmed that these plants were genetically similar and can be used as elite plants.

Journal ArticleDOI
TL;DR: None of the treatments including BA or kinetin alone or BA in combination with NAA produced significantly more shoots for commercial exploitation, and PCR amplification with 14 different random primers confirmed no somaclonal variant up to two cycles of shoot multiplication.
Abstract: 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 (01 mg l−1) or kinetin (05 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 01 mg l−1 BA and 05 mg l−1 kinetin Shoot proliferation decreased with increasing concentration of BA alone, but in case of a combination of BA and NAA (01 mg l−1 each), it increased with increasing concentration of BA up to 100 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 (55 shoots per explants) proliferation was observed on MS medium containing 10 mg l−1 BA + 05 mg l−1 kinetin + 05 mg l−1 IBA or 025 mg l−1 BA + 10 mg l−1 kinetin + 10 mg l−1 IBA Regenerated shoots showed root induction on MS basal medium or on MS medium containing 10 mg l−1 IBA It is concluded that a five-fold increase (10 mg l−1 BA + 05 mg l−1 kinetin + 05 mg l−1 IBA) in axillary shoot proliferation, while seven-fold increase (025 mg/l mg l−1 BA + 10 mg l−1 kinetin + 10 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

Journal Article
TL;DR: A study on the effects of two growth regulators, 2, 4-D (2,4-dichlorophenoxyacetic acid) and kin (kinetin), in concentrations of 0, 0.5, 1.0 and 2.0 mg l -1 each in a factorial design, as a possible means of enhancing the growth rates of Haematococcus pluvialis and Dunaliella salina.
Abstract: : Haematococcus pluvialis Flotow and Dunaliella salina Teodoresco are commercially important because of their ability to accumulate very high carotenoid contents. However, their use is hindered by their slow growth rates. This paper reports a study on the effects of two growth regulators, 2,4-D (2,4-dichlorophenoxyacetic acid) and kin (kinetin), in concentrations of 0, 0.5, 1.0 and 2.0 mg l -1 each in a factorial design (2 4 combinations), as a possible means of enhancing the growth rates. After 12-13 days of treatment with plant hormones, D. salina showed a significant increase in growth with all the hormone concentrations and combinations used and under 15% salinity (NaCl, w/v), (except for 0.5 mg l -1 2,4-D and no kin), with up to 410% more cells than the control; under 10% salinity (NaCl, w/v), the increase in growth was significant with 0.5 mg l -1 2,4-D and no kin (180% more cells than the control), and also with 1.0 mg l -1 2,4-D and no kin (126% more cells than the control) and 2.0 mg l -1 2,4-D and 0.5 mg l -1 kin (134% more cells than the control) in the culture medium. Cultures of H. pluvialis were significantly influenced under 1.0 mg l -1 2,4-D (with 320% more cells than the control), but also showed a significant increase in the growth rate when the ratio auxin to cytokinin was 1 (equal concentrations of 1.0 mg l -1 of both growth regulators) with more than 290% cells than the control, and with 0.5 mg l -1 2,4-D and 2.0 mg l -1 kin (200% more cells than the control) in the culture medium.

01 Jan 2013
TL;DR: The present study was to investigate the best plant growth regulators for shoot proliferation and multiplication; and to determine the optimum concentrations of phytohormones for shoot tip culture of banana cv.
Abstract: The present study was to investigate the best plant growth regulators for shoot proliferation and multiplication; and to determine the optimum concentrations of phytohormones for shoot tip culture of banana cv. Agishwar by observing the effect of different culture conditions during induction. A micro- propagation protocol for banana ( Musa sp.) cv. Agnishwar was established by using shoot tip culture. Shoot tips obtained by removing leaf sheaths from sucker were cultured aseptically in MS (Murashige and Skoog) medium supplemented with different concentrations of cytokines viz. 6-benzylaminopurine (BAP), kinetin (kin), N 6 - (2-isopentyl) adenine (2iP) for multiplication of shoot and auxins viz. Indole-3- butyric acid (IBA), α α α α-naphthalene acetic acid (NAA) for induction of root. Maximum multiplication (95%) was obtained in MS medium containing 4.0 mg/1 BAP. The highest average number of shoots for each explant (5.9) was found in MS medium fortified with 4.0 mg/l BAP while maximum elongation of shoot (4.9cm) was observed in MS medium having 5.0 mg/l BAP. IBA at a concentration of 1.0 mg/l was found most suitable for rooting of shoot. The rooted shoots were acclimatized and successfully transferred to plastic pots. After hardening, they were transferred to the main field and the survival rate was around 90%.This protocol might be used for the massive in vitro production of the plantlets of banana cv. Agnishwar.

Journal ArticleDOI
TL;DR: A protocol for somatic embryogenesis of hybrid C. arabica from leaf explants is described here, which will enable this crop to multiply and supply large numbers of high quality hybrid coffee seedlings to farmers engaged in coffee production.
Abstract: Summary Coffea arabica is one of the most traded agricultural commodities in the World, and the major export commodity of Ethiopia. Improvement of this crop requires the development of hybrid varieties that possess desirable traits. The production and distribution of hybrid coffee is difficult due to the high costs required for manual crossing and maintenance. The use of somatic embryogenesis is an effective means of propagation. A protocol for somatic embryogenesis of hybrid C. arabica from leaf explants is described here. Leaf explants collected from healthy 1-year-old seedlings were used. The highest percentage of callus induction was observed from explants cultured on MS medium containing 1.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 2.0 mg l–1 6-benzylaminopurine (BAP). Embryogenic calli were obtained from leaf explants cultured on MS medium supplemented with 0.05 mg l–1 kinetin in combination with 0.1 mg l–1 indole-3-butyric acid (IBA). The highest number of embryos that germ...

Journal ArticleDOI
01 Dec 2013
TL;DR: An improved micropropagation protocol was developed for Zediber moran and Z. zerumbet, two wild species of the genus Zingiber, found in Northeast India, and found to be the optimum for shoot multiplication and regeneration.
Abstract: 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.

Journal ArticleDOI
TL;DR: Results described here confirm the reliability of this protocol for micropropagation and delivery of desirable gene using A. tumefaciens into indica rice and show no significant variation in the 2C DNA content and Ploidy level between wild type IR36 and in vitro maintained rice lines.
Abstract: Cereal crops are the major targets for transformation mediated crop improvement and IR36 is an early maturing, high yielding, insect and disease resistant rice variety however, it is abiotic stress sensitive. Hence, development of an efficient and reproducible micropropagation system via somatic embryogenesis and Agrobacterium tumefaciens mediated transformation is prerequisite to develop abiotic stress tolerant IR36. Further, Genetic stability of analysis of plantlets through RAPD and ISSR and Ploidy level through Flow cytometry (FCM) measurement of 2C DNA content is necessary for future application of transformed IR36. In this study, Mature seeds inoculated on (Murashige and Skoog) MS medium with 11.31 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.3 μM Kinetin (Kn) had highest callus induction frequency (98%). The highest regeneration frequency (80%) was observed in MS + 13.28 μM Benzyladenine (BA) with 8.06 μM α-naphthalene acetic acid (NAA). Randomly Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Flow Cytometry (FCM) analysis showed no significant variation in the 2C DNA (0.81 pg/2C) content and Ploidy level between wild type IR36 and in vitro maintained rice lines. Of the various OD bacterial culture, an optimum OD of 0.4 and inoculation duration of 10 min resulted in efficient Agrobacterium-mediated transformation. β-glucuronidase activity was maximum in callus (99.05%). These results described here confirm the reliability of this protocol for micropropagation and delivery of desirable gene using A. tumefaciens into indica rice.

Journal ArticleDOI
TL;DR: Higher density of veins and glandular trichomes was observed in the leaves of hardened plants, and stomata became functional during hardening which were non-functional under in vitro condition.
Abstract: We developed a micropropagation protocol for Cleome gynandra, a C4 model plant with medicinal importance. Surface-sterilized nodal segments obtained from 1 to 2-month-old field grown plant were used as explants for culture establishment and plant regeneration. Multiple shoots differentiated through bud breaking on Murashige and Skoog (MS) medium with different concentrations of benzyladenine (BA) and kinetin (Kin). The optimum shoot differentiation occurred on medium with 1.5 mg l−1 BA. Out of various concentrations and combinations of cytokinins and auxins, MS medium containing 0.5 mg l−1 BA and 0.1 mg l−1 IAA (indole-3-acetic acid) was found best for shoot multiplication. However, the differentiated shoots exhibited hyperhydration, leaf curling and early leaf fall during subculturing. To overcome these problems, regenerated shoots were transferred to the modified MS medium with reduced nitrates (825 mg l−1 NH4NO3 and 950 mg l−1 KNO3) and 100 mg l−1 (NH4)2SO4. The micropropagated shoots were rooted (i) in vitro on one-fourth strength of MS salts with 0.25 mg l−1 each of IBA (indole-3 butyric acid) and NOA (2-naphthoxyacetic acid) + 100 mg l−1 activated charcoal, and (ii) ex vitro, by treating the shoot base(s) with 200 mg l−1 of IBA for 3 min and transferred to soilrite moistened with one-fourth strength of MS macro salts in culture bottles. The plants were hardened in the greenhouse with 85 % survival rate. Micromorphological studies of the plants were conducted during hardening with reference to development and changes in vein spacing, glandular trichome and stomata. In comparison to leaves under in vitro condition, higher density of veins and glandular trichomes was observed in the leaves of hardened plants. In addition, stomata became functional during hardening which were non-functional under in vitro condition.

Patent
20 Mar 2013
TL;DR: In this paper, a method for improving transplanting survival rate of rhizoma bletillae tissue culture seedling was proposed, which is capable of inducing anti-adversity of the seedling during the root culture.
Abstract: The invention discloses a method for improving transplanting survival rate of rhizoma bletillae tissue culture seedling. The special culture medium comprises 1/2 MS culture medium +6-butyl acrylate (BA) 1.0 mg/l + naphathyl acetic acid (NAA) 0.75 mg/l + coconut milk 20 percent + cane sugar g/l + agar power 8g/l, enrichment medium MS culture medium + 6-BA 1.0 mg/l + kinetin (KT) 3.0 mg/l + cane sugar 30g/l + agar powder 8g/l, and root medium MS + NAA 0.5 mg/l + cane sugar 20 g/l + agar powder 8g/l. The steps are: A. germinating rhizoma bletillae seeding under a germ-free condition; B. conducting the enrichment culture; C. conducting root culture and induction of anti-adversity; D. conducting acclimatization and transplant. As self-control resistance inducer 10m/l of sialic acid (SA) 0.3 g/l + cathepsin (CTS) 2g/l is added during the period of the root culture, the method for improving the transplanting survival rate of the rhizoma bletillae tissue culture seedling is capable of inducing anti-adversity of the tissue culture seedling during the root culture, increasing adaptive capacity to the outer environment, and improving transplanting survival rate of the tissue culture seedling.

Journal ArticleDOI
TL;DR: Cytological and random amplified polymorphic DNA (RAPD) analyses were carried out to assess the genetic integrity of the regenerated plantlets, suggesting genetic stability in the micropropagated plants of N. khasiana.
Abstract: An efficient in vitro protocol for large-scale multiplication of Nepenthes khasiana, a threatened insectivorous plant of India, has been developed from nodal stem segments. The highest shoot proliferation of 19.16 ± 0.23 shoots/explant was recorded in half-strength Murashige and Skoog (MS) medium supplemented with 2.5 mg/l kinetin, 2.0 mg/l 6-benzyl aminopurine, 3 % sucrose and 0.8 % agar. The best rooting was achieved in half-strength MS medium supplemented with 2.0 mg/l α-naphthalene acetic acid with an average of 9.04 ± 0.46 roots/shoot. The plantlets were successfully transferred to the greenhouse with survival rate of 92 %, exhibiting normal development. Cytological and random amplified polymorphic DNA (RAPD) analyses were carried out to assess the genetic integrity of the regenerated plantlets. Cytological analysis revealed no change in chromosome number with cells studied showing 2n = 80. Of the 80 primers screened for RAPD analysis, 14 primers resulted in clear and scorable bands. A total of 72 amplification products were obtained out of which only 4.1 % bands were polymorphic. Cluster analysis of the RAPD profile revealed an average similarity coefficient ranging from 0.98 to 1.0, thus suggesting genetic stability in the micropropagated plants of N. khasiana.