Showing papers on "Kinetin published in 2011"
TL;DR: Of the two elicitors, SA was more effective in stimulating the accumulation of hypericins and pseudohypericin in shoot cultures of Hypericum hirsutum and H. maculatum, and it is suggested that culture of shoots on MS medium supplemented with BA or Kin enhanced production ofhypericins in H. Maculatum and hyperforin inH.
Abstract: We investigated the effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 6-γ,γ-dimethylallylaminopurine (2iP), thidiazuron (TDZ) and α-naphthaleneacetic acid (NAA)], modified Murashige and Skoog (MS) medium containing 10 mM NH4
+ and 5 mM NO3
− and supplemented with 2iP, BA, Kin and NAA (MSM medium), and two elicitors [jasmonic acid (JA), and salicylic acid (SA)], on plant growth and accumulation of hypericins (hypericin and pseudohypericin) and hyperforin in shoot cultures of Hypericum hirsutum and H. maculatum. Our data suggested that culture of shoots on MS medium supplemented with BA (0.4 mg l−1) or Kin (0.4 mg l−1) enhanced production of hypericins in H. maculatum and hyperforin in H. hirsutum. Hypericins and hyperforin concentrations decreased in both species when TDZ (0.4 mg l−1) was added to the MS medium. Also, TDZ induced hyperhydric malformations and necrosis of regenerated shoots. Cultivation of H. maculatum on MSM medium resulted in approximately twofold increased production of hypericins compared to controls, and the growth of H. hirsutum shoots on the same medium led to a 6.16-fold increase in hyperforin production. Of the two elicitors, SA was more effective in stimulating the accumulation of hypericins. At 50 μM, SA enhanced the production of hypericin (7.98-fold) and pseudohypericin (13.58-fold) in H. hirsutum, and, at 200 μM, enhanced the production of hypericin (2.2-fold) and pseudohypericin (3.94-fold) in H. maculatum.
203 citations
TL;DR: This experiment suggests that glucose and fructose may play important roles during salt stress because there were clearly higher concentrations of these sugars in the grain of the stressed cultivars under IAA and KIN application: it appears that their accumulation was the earliest response detected during the grain-filling period in rice.
Abstract: Phytohormones, such as auxin and cytokinin, are known to be involved in the regulation of plant responses to salinity stress and counteract the adverse effect of stress conditions. This work investigated the effects of the exogenous spraying of indole-3-acetic acid (IAA) and kinetin (KIN) during the reproductive phase on grain yield by examining the 1000-grain weight and filled-grain percentage as well as the changes in starch, total soluble sugars, sucrose, glucose and fructose concentrations in the grains of two rice cultivars under salt stress. The results indicated that the applied IAA and KIN led to an increased grain yield, 1000-grain weight and filled-grain percentage for both rice cultivars under salt stress. The storage starch content in the grain of the salt-sensitive cultivar was more than that in the salt-tolerant cultivar under IAA application compared with KIN, whereas a decrease in the total soluble sugar content was observed with both IAA and KIN treatments, in comparison to the non-hormone treatment. Interestingly, this study showed that IAA led to a much higher increase in the sucrose content in grain, as compared to the KIN. Furthermore, this experiment suggests that glucose and fructose may play important roles during salt stress because there were clearly higher concentrations of these sugars in the grain of the stressed cultivars under IAA and KIN application: it appears that their accumulation was the earliest response detected during the grain-filling period in rice. Finally, this work indicated that an increase in the rice grain yield, 1000-grain weight and filled-grain percentage are associated with an increase in the contents of starch, sucrose, glucose and fructose in grain caused by the application of IAA and KIN.
101 citations
91 citations
TL;DR: The developed micropropagation protocol has the potential for producing more than 60,000 transplantable shoots per year from a single shoot-tip explant of this critically endangered plant with horticultural potential.
Abstract: Low shoot multiplication, morphological abnormalities, poor rooting frequency and high cost of production are among the factors challenging the micropropagation of ornamental perennials and garden plants. Most of these problems can be alleviated by using the appropriate type and concentration of plant growth regulator(s) (especially cytokinins) in developing efficient micropropagation protocols. In this study, we investigated the effects of five different aromatic cytokinins (BA, Kin, mT, mTR and MemTR) on adventitious shoot production from shoot-tip explants of B. greenii, a critically endangered plant with horticultural potential. Of all the cytokinin concentrations evaluated, the highest adventitious shoot production (5.88 ± 0.73 shoots/explant) was observed in cultures containing 7 μM MemTR. Low adventitious shoot production, which was not significantly different from that of the control, was observed at all the concentrations of kinetin (Kin), suggesting that it is a weak cytokinin for shoot production in this species. All the treatments with BA alone showed higher adventitious shoot production when compared to the BA treatments supplemented with NAA concentrations. At equimolar concentrations, however, all the BA concentrations had a higher abnormality index than the other cytokinins. It is noteworthy that the abnormality index in all the topolin treatments was much lower than that recorded at the lowest BA concentration. Almost all the abnormality indices recorded with mTR and MemTR concentrations were lower than that of the control. Given that the explants used were from BA-containing cultures, it is likely that the abnormalities recorded using mTR and MemTR were carry-over effects of BA. Culturing under 16 h light/8 h dark conditions resulted in a higher production of adventitious shoots with lengths greater than 10 mm compared to culturing under continuous light. This measure could help reduce the cost of production. Regenerated shoots were successfully rooted and acclimatized with a 65% survival frequency and no observable morphological variation. The developed micropropagation protocol has the potential for producing more than 60,000 transplantable shoots per year from a single shoot-tip explant of this critically endangered species.
88 citations
01 Apr 2011
TL;DR: In plant tissue culture, growth regulator has significant roles such as to control root and shoot development in the plant formation and callus induction and the existence of a certain growth regulating substances can enhance growth regulator activity of other substances.
Abstract: The Role of Growth Regulator in Tissue Culture Plant Propagation. Endang G. Lestari. In plant tissue culture, growth regulator has significant roles such as to control root and shoot development in the plant formation and callus induction. Cytokinin and auxin are two prominent growth regulator. Cytokinin consists of BA (benzil adenin), kinetin (furfuril amino purin), 2-Ip (dimethyl allyl amino purin), and zeatin. While auksin covers IAA (indone acetic acid), NAA (napthalene acetic acid), IBA (indole butiric acid) 2.4-D (2.4- dicholophenoxy acetic acid), dicamba (3,6 dicloro-O-anisic acid), and picloram (4-amino 3,5,6-tricloropicolinic acid). The emphasis of plant growth purposes decide the use of growth regulator. Cytokinin is applied mainly for the purpose of shoot, while auxin is mainly used for the purpose of root and callus. The application of growth regulator application is varied, depending on the genotype and physiological condition of the plant. The existence of a certain growth regulating substances can enhance growth regulator activity of other substances. The type and concentration of the appropriate growth regulators for each plant is not the same because it depends on the genotype and physiological condition of plant tissue. However so often both are frequently required depend on the ratio/ratio of auxin cytokines or vice versa. The existence of a certain growth regulating substances can enhance growth regulator activity of other substances. The type and concentration of the appropriate growth regulators for each plant is not the same because it depends on the genotype and physiological condition of plant tissue. For the propagation, multiple and adventive shoots along with embriosomatic formation could be applied. The seedling is obtained from one somatic cell. Here, strong auxin, such as dicamba and picloram 2.4-D, is utilized for callus production. For this reason, seedling per unit could be produced more than that of organogenesis.
84 citations
TL;DR: In this study, the propagation of P. rothschildianum was achieved through the in vitro formation of secondary protocorm-like bodies (PLBs) from the primary PLB that developed from stem-derived callus.
Abstract: Paphiopedilum orchids are among the world’s most popular orchid due to their impressively beautiful flowers. Propagation of these orchid genera has been hampered by the naturally slow growth rate of the plant, which renders it very difficult to be propagated through conventional methods. In vitro culture techniques have provided a useful alternative technology for propagating this recalcitrant species. In this study, the propagation of P. rothschildianum was achieved through the in vitro formation of secondary protocorm-like bodies (PLBs) from the primary PLB that developed from stem-derived callus. The PLBs were cultured on half-strength MS medium supplemented with different concentrations (1.0, 2.0, 3.0, and 4.0 μM) of 6-benzyladenine (BA) and kinetin for the induction of secondary PLBs. The highest number of secondary PLBs formed was obtained on half-strength MS medium supplemented with 4.0 μM kinetin, with an average of 4.1 PLBs per explant after 8 weeks of culture. The secondary PLBs continued to proliferate further and formed 9.5–12.1 new PLBs per secondary PLB after being subcultured onto half-strength plant growth regulator-free MS medium supplemented with 60 g/L banana homogenate (BH). These tertiary PLBs were subcultured onto media containing different organic additives, such as BH, coconut water, potato homogenate, and tomato homogenate, for plantlet regeneration. Among the organic additives tested, the addition of 20% CW to half-strength MS medium resulted in the best average plantlet regeneration percentage from the PLBs, 67.9%, after 8 weeks of culture.
81 citations
TL;DR: The highest level of rutin was produced when adventitious roots were grown in a 34/66 AM/NI full-strength standard MS medium containing 5 mg/l IAA, and different ammonium/nitrate (AM/NI) ratios in a root suspension culture also greatly affected r Rutin production and its secretion into a liquid medium.
Abstract: Rutin is an economically valuable flavone compound with anticancer activity, dietary effects, and anti-aging activity. In this study, callus and adventitious roots were induced from three Morus (mulberry) species. Among the three mulberry species tested for rutin production, roots of the Sugye (M.alba L.) had the highest levels (242.2 μg/g fresh tissue) of rutin. In addition, the mature leaves of this type of tree promoted higher levels of rutin compared to those of young leaves or those undergoing senescence. Adding auxins such as indole-3-acetic acid (IAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthalene-1-acetic acid (NAA) not only enhanced the development of callus and adventitious roots but also increased the protein and rutin contents. In contrast, adding cytokinins such as 6-benzyladenine (BA) and kinetin (KN) retarded callus and adventitious root development as well as the protein and rutin contents. Callus in suspension culture in the presence of IAA produced more rutin than that in the absence of IAA. However, rutin secretion into a medium was greater in the absence of IAA. Different ammonium/nitrate (AM/NI) ratios in a root suspension culture also greatly affected rutin production and its secretion into a liquid medium. As a result, the highest level of rutin was produced when adventitious roots were grown in a 34/66 AM/NI full-strength standard MS medium containing 5 mg/l IAA.
80 citations
TL;DR: Not only did kinetin produce the desired effect on splicing in FD patients but also that effect seems to improve with time despite lack of dose change, the first report of a drug that produces in vivo mRNA splicing changes in individuals with FD.
Abstract: 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 WT IKBKAP mRNA and IKAP protein expression in FD cell lines and carriers. To determine whether 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/d for 28 d. An increase in WT IKBKAP mRNA expression in leukocytes was noted after 8 d in six of eight individuals; after 28 d, the mean increase compared with 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 seems 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 whether kinetin will prove therapeutic in FD patients.
74 citations
Journal Article•
TL;DR: In this paper, the stimulatory effect of kinetin (20 and 40 ppm), ascorbic acid and glutamic acid (100 and 200 ppm) on growth and chemical constituents of Codiaeum variegatum L. was investigated.
Abstract: 2 Abstract: Two pot experiments were conducted during two successive seasons (2009 and 2010) in the nursery of the National Research Center. The aim of this work is to study the stimulatory effect of kinetin (20 and 40 ppm), ascorbic acid (100 and 200 ppm) and glutamic acid (100 and 200 ppm) on growth and chemical constituents of Codiaeum variegatum L. Results showed that, increasing concentration of the three foliar applications gradually increased all growth parameters (plant height, number of branches, number of leaves, stem diameter, root length as well as fresh and dry weights of all plant organs) and also the content of the total carbohydrates, nitrogen, phosphorus and potassium percentages. The effect of glutamic acid was superior to that of kinetin, ascorbic acid on increasing plant growth at vegetative growth especially when plants were sprayed with glutamic acid at 200 ppm.
59 citations
TL;DR: A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed using nodal segments of fresh shoot sprouts originated from axillary buds obtained from a plant around 35–40 years old lopped plant for establishment of in vitro cultures.
Abstract: A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed. Nodal segments of fresh shoot sprouts originated from axillary buds obtained from a plant around 35–40 years old lopped plant were used as explants for establishment of in vitro cultures. Surface-sterilized explants produced optimum number of shoots through activation of axillary buds on Murashige and Skoog’s (MS) medium containing 8.88 μM BA (6-benzyladenine) + additives (25 mgl−1 each of adenine sulphate, arginine, citric acid, 50 mgl−1 ascorbic acid). The shoot multiplication was influenced by the successive transfer of the mother explants for 4–5 passages. The maximum number (23.1 ± 0.73 shoots per explant) of shoots were regenerated on MS supplemented with 1.11 μM BA + 1.16 μM Kn (Kinetin) + 0.54 μM NAA (α-naphthalene acetic acid). About 90% shoots pulse-treated with a combination of 2460.27 μM Indole-3-butyric acid (IBA) + 494.56 μM NOA (2-naphthoxy acetic acid) were rooted ex vitro on soilrite within 15–18 days. Over 80% cloned plantlets were hardened successfully in a green house and transferred to polybag/pots.
55 citations
TL;DR: Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied and rooting response was reduced in non-toxic genotype as compared to toxic genotypes.
TL;DR: 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).
TL;DR: An efficient and cost effective micropropagation protocol using liquid medium was developed for Catharanthus roseus, a commercially important medicinal plant, and lays the foundations for the shifting of plant production from small to commercial scale.
Abstract: An efficient and cost effective micropropagation protocol using liquid medium was developed for Catharanthus roseus, a commercially important medicinal plant. Comparative analysis of shoot growth and proliferation in liquid Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins [6-Benzyladenine (BA), Kinetin (KN) and Thidiazuron (TDZ)] was conducted. Better response in terms of shoot proliferation, shoot diameter, number of leaves/shoot, number of branches/shoot, fresh weight and dry weight was observed in a liquid medium vis-a-vis solid medium. A sample of 20 ml of liquid medium supplemented with 5 μM of BA was optimized for propagation of C. roseus by a liquid culture system. Among various concentrations of auxins tried, 1-Naphthaleneacetic acid (NAA) 5 μM was found to be the best for root induction. Quantification of pharmaceutically important constituents (vincristine and vinblastine) and total alkaloid content of microshoots grown in solid and liquid medium as well as in vitro raised plants and mother plant was also conducted, hitherto unreported in this high-value medicinal plant. This work further lays the foundations for the shifting of plant production from small to commercial scale.
TL;DR: Genetic variations of field-established B. nutans plants regenerated through tissue cultures revealed a high level of genetic stability, and February–March and December were found to be the best seasons for culture establishments.
Abstract: Bambusa nutans Wall., is an evergreen, perennial, and multipurpose bamboo having strong culms, which are largely used for construction, scaffolding, craft purposes, pulp, and paper industry. Multiple shoots from nodal segments (3–4 cm) of young branches of mature culms were established in Murashige and Skoog (1962) (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) (1.0–6.0 mg l−1) or in combination with α-naphthaleneacetic acid (NAA) (0.5–1.0 mg l−1) or kinetin (Kn) (1.0–2.0 mg l−1). February–March and December were found to be the best seasons for culture establishments. Maximum shoots were achieved on MS medium fortified with BAP (2.0 mg l−1). Embryogenic callus (slightly greenish compact, globular, and slow growing) was initiated from the base of severed sprouted buds in 2–3 subsequent subcultures on MS medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (5.0 mg l−1) under dark incubations. Maturation and germination of well-organized somatic embryos was achieved on MS medium containing BAP and 2,4-D (1.0 mg l−1 each) with 20.0 mg l−1 ascorbic acid. Full-strength MS medium supplemented with 2% glucose favored further development of proliferated somatic embryos into plantlets. Genetic variations of field-established B. nutans plants regenerated through tissue cultures were assessed by amplified fragment length polymorphism (AFLP) analysis using 6 primer combinations. Four hundred and seven scorable fragments were amplified, of which 402 (98.8%) have recorded conservation at various morphogenetic stages leading to plantlets regeneration, therefore, revealed a high level of genetic stability.
TL;DR: An efficient in vitro propagation protocol, applicable both to young and mature explants of two Thymus spp.
Abstract: An efficient in vitro propagation protocol, applicable both to young and mature explants of two Thymus spp., results in genetically stable plantlets. In vitro-grown shoot tips of Thymus vulgaris L. were exposed to cytokinins (6-benzyladenine, kinetin, and thidiazuron) alone or in combination with auxins, gibberellic acid (GA3) and/or silver nitrate in order to optimize in vitro shoot proliferation. Optimum shoot proliferation (97% regeneration rate, with 8.6 shoots produced per explant) was obtained when semi-solid Murashige and Skoog (MS) medium was supplemented with 1 mg L−1 kinetin and 0.3 mg L−1 GA3. Rooting of the shoots was easily obtained on semi-solid MS medium that was either hormone-free or supplemented with auxins. However, the best root apparatus (92.5% rooting rate, with 19 adventitious roots per shoot) developed on MS medium supplemented with 0.05 mg L−1 2,4-dichlorophenoxyacetic acid. Genetic stability was confirmed in the in vitro-germinated mother plant as well as the shoots that underwent two, four, six, eight, or ten cycles of in vitro subculturing by random amplified polymorphic DNA (RAPD) analysis. When applied to the micropropagation of mature shoot tips of T. longicaulis C. Presl subsp. longicaulis var. subisophyllus (Borbas) Jalas, the optimized in vitro propagation protocol resulted in a 97.5% shoot regeneration rate, with five shoots formed per explant, and 100% rooting. Rooted plantlets of both species were transferred to 250-mL plastic pots and successfully acclimatized by gradually reducing the relative humidity.
TL;DR: A reproducible protocol for direct and indirect somatic embryogenesis was established in a small aromatic tree and the initiation, development, and germination of somatic embryos from both explants were confirmed.
Abstract: A reproducible protocol for direct and indirect somatic embryogenesis was established in a small aromatic tree, Murraya koenigii. Embryogenic callus was obtained from 90% zygotic embryonic axis (ZE) and 70% cotyledon (COT) explants in Murashige and Skoog (MS) basal medium supplemented with 8.88 μM 6-benzyladenine (BA) and 2.675 μM α-naphthaleneacetic acid (NAA). Globular somatic embryos were induced and further matured from such embryogenic callus by subsequent culture on the same basal media containing thidiazuron (TDZ) (2.27–9.08 μM). The highest frequency of somatic embryos (14.58 ± 0.42) was recovered from ZE-derived callus after 6 weeks. The age and type of explant and concentration of TDZ played an important role in the development of somatic embryos. Explants excised from 60-day-old seed differentiated from 96.67% of ZE explants and 86.67% from COT explants when cultured on MS basal medium supplemented with 4.54 and 9.08 μM TDZ, respectively, after 4 weeks. The best result obtained for the average frequency of somatic embryos (11.28 ± 0.32) was from ZE explants, which was significantly higher than COT explants (7.34 ± 0.97). Most of the somatic embryos (above 95%), irrespective of their origin, germinated after 4 weeks in 1/2 MS basal media containing 2.32 μM kinetin (KN) and 1.07 μM NAA. Well-rooted plantlets were successfully acclimatized. Histological analysis and scanning electron micrographs confirmed the initiation, development, and germination of somatic embryos from both explants.
TL;DR: In this paper, a comparative study was carried out with ethanolic (80%) extracts from leaves and cell cultures of three Callistemon species, namely C. lanceolatus (CL), C. viridiflorous (CV), and C. comboynensis (CC).
Abstract: A comparative study was carried out with ethanolic (80%) extracts from leaves and cell cultures of three Callistemon species, namely C. lanceolatus (CL), C. viridiflorous (CV), and C. comboynensis (CC). Cell suspensions of the three species were grown in liquid Murashige and Skoog (MS) medium (100 ml) supplemented with 0.9 mg·g-1 kinetin in combination with 1.1 mg·g-1 NAA. The CL leaf extract was standardized to contain the highest amount of phenolics (104 ± 2.0 mg·g-1), followed by CC (95.8 ± 1.2 mg·g-1) and CV (79.8 ± 4.6 mg·g-1). On the other hand, cell cultures of CV contained more phenolics (14.9 ± 0.6 mg·g-1) than those of the other two species, CL and CC, which contained 12.2 ± 0.16 and 9.12 ± 0.16 mg·g-1, respectively. Nevertheless, CV leaf extract exhibited the highest antioxidant activity (91.4% ± 0.4%) at a concentration of 1000 µg·ml-1, comparable to 100 µg·ml-1 gallic acid (90.8% ± 1.5%).
TL;DR: The histological sections at different developmental stages of shoot buds revealed the organization of nodular meristematic zone leading to the orientation and differentiation ofshoot buds in large number and thereafter conversion into healthy shoots.
Abstract: The aim of this study was to develop a new micropropagation system for Cassia angustifolia Vahl., an important medicinal legume using root explant as starting material. Root explants taken from 30-day-old aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators: 6-benzyladenine (BA), kinetin (Kn), and thidiazuron (TDZ). Organogenic nodular calli obtained on MS + TDZ (1.0 μM) were transferred to shoot regeneration medium supplemented with different cytokinins (BA, Kn or TDZ) either alone or in combination with auxin:indole-3-acetic acid or α-naphthalene acetic acid. Maximum shoot regeneration frequency (90%) was obtained on MS + BA (2.5 μM) + NAA (0.6 μM) wherein a maximum of 42.76 ± 1.47 shoot buds per explant were induced with a maximum conversion rate of 35.63 ± 0.75 shoots per explant and average shoot length of 5.43 ± 0.20 cm. Elongated microshoots were successfully rooted under ex vitro conditions by pulse treatment in 200 μM of indole-3-butyric acid for half an hour. Microshoots were rooted, acclimatized and hardened off simultaneously in sterilized soilrite inside the growth room and then established in pots containing sterilized soil and manure (1:1) and grown under greenhouse condition with 90% survival rate. The histological sections at different developmental stages of shoot buds revealed the organization of nodular meristematic zone leading to the orientation and differentiation of shoot buds in large number and thereafter conversion into healthy shoots.
TL;DR: An efficient method was developed to multiply nodal explants of NAG using liquid-culture medium and a simple temporary immersion culture vessel and may be useful in providing NAG tissues or plants for a range of ginseng-based natural health products.
Abstract: North American ginseng (NAG) (Panax quinquefolius L.) is a medicinally important plant with multiple uses in the natural health product industry. As seed propagation is time-consuming because of the slow growth cycle of the plant, in vitro propagation using a bioreactor system was evaluated as an effective approach to accelerate plant production. An efficient method was developed to multiply nodal explants of NAG using liquid-culture medium and a simple temporary immersion culture vessel. The effects of plant growth regulators, phenolics, and chemical additives (activated charcoal, melatonin, polyvinylpolypyrrolidone, and ascorbic acid) were evaluated on in vitro-grown NAG plants. The highest number (12) of shoots per single node was induced in half-strength Schenk and Hildebrandt basal medium containing 2.5 mg/l kinetin, in which 81% of the cultured nodes responded. In a culture medium with 0.5 mg/l α-naphthalene acetic acid (NAA), roots were induced in 78% of the explants compared to 50% with a medium containing indole-3-acetic acid. All of the resulting plants appeared phenotypically normal, and 93% of the rooted plants were established in the greenhouse. Phenolic production increased significantly (P < 0.05) over a 4-wk culture period with a negative impact on growth and proliferation. Activated charcoal (AC; 50 mg/l) significantly reduced total phenolic content and was the most effective treatment for increasing shoot proliferation. Shoot production increased as the phenolic content of the cultures decreased. The most effective treatment for NAG development from cultured nodal explants in the bioreactor was 2.5 mg/l kinetin, 0.5 mg/l NAA, and 50 mg/l AC in liquid culture medium. This protocol may be useful in providing NAG tissues or plants for a range of ginseng-based natural health products.
TL;DR: Direct regeneration via leaf explants will be useful for Agrobacterium-mediated genetic transformation, and will facilitate pathway manipulation using metabolic engineering for bioactive withanolides.
Abstract: A micropropagation system through leaf explant culture has been developed for Withania coagulans. Shoot bud proliferation occurred through both adventitious and de novo routes depending on the hormonal regime of the culture medium. Green compact nodular organogenic callus developed on Murashige and Skoog (MS) medium supplemented with 2.3 μM kinetin (Kn) and lower levels of 6–benzyladenine (BA) (13.3 μM) while multiple adventitious shoot bud differentiation occurred on medium fortified with 2.3 μM kinetin (Kn) and higher levels of BA (22.2 μM). Shoot buds were transferred to proliferation medium containing 2.2 μM BA, 2.3 μM Kn, and 3.9 μM phloroglucinol (PG) for further growth and development of shoot system. Elongated shoots were rooted using a two-step procedure involving pulse treatment of 7 days in a medium containing 71.6 μM choline chloride (CC) and 3.9 μM PG and then transferred to rooting medium containing ½ MS, 1.2 μM IBA, 3.6 μM PAA, and 14.3 μM CC for 3 weeks. Well-rooted plants were transferred to a greenhouse for hardening and further growth. Random amplification of polymorphic DNA (RAPD) showed monomorphic bands in all the plants thereby confirming clonality of the regenerants. Thin layer chromatography (TLC) showed the presence of withanolides in the regenerated plants. Quantification through reverse-phase HPLC revealed increased concentration of withanolides in the regenerated plants compared to the field-grown mother plant. Accumulation of withaferin A and withanolide A increased up to twofold and that of withanone up to tenfold. Direct regeneration via leaf explants will be useful for Agrobacterium-mediated genetic transformation, and will facilitate pathway manipulation using metabolic engineering for bioactive withanolides.
TL;DR: Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages and an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans is described.
Abstract: This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with 4.4 μM benzylaminopurine (BA) and 2.32 μM kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with 13.2 μM BA, 2.32 μM Kin, and 0.98 μM indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with 9.8 μM IBA, 2.85 μM indole-3-acetic acid (IAA), 2.68 μM naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.
TL;DR: Mature seed-derived embryogenic calli of indica rice induced on semisolid Murashige and Skoog medium showed normal growth while the non-transgenic control plantlets turned yellow and finally did not survive, and in vitro screening for salt tolerance showed increase in fresh mass of OsglyII putative transgenic calli.
Abstract: Mature seed-derived embryogenic calli of indica rice (Oryza sativa L. cv. PAU201) were induced on semisolid Murashige and Skoog medium supplemented with 2.5 mg dm -3 2,4-dichlorophenoxyacetic acid + 0.5 mg dm -3 kinetin + 560 mg dm -3 proline + 30 g dm -3 sucrose + 8 g dm -3 agar. Using OsglyII gene, out of 3180 calli bombarded, 32 plants were regenerated on medium containing hygromycin (30 mg dm -3 ). Histochemical GUS assay of the hygromycin selected calli revealed GUS expression in 50 % calli. Among the regenerants, 46.87 % were GUS positive. PCR analysis confirmed the presence of the transgene of 1 kb in 60 % of independent plants. Further, these plants have been grown to maturity in glasshouse. In vitro screening for salt tolerance showed increase in fresh mass of OsglyII putative transgenic calli (185.4 mg) as compared to control calli (84.2 mg) on 90 mM NaCl after 15 d. When exposed to 150 mM NaCl, OsglyII putative transgenic plantlets showed normal growth while the non-transgenic control plantlets turned yellow and finally did not survive.
TL;DR: Genetic stability of micropropagated clones were periodically evaluated at an interval of 6 months up to 30 months in culture using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis and genetic uniformity in all regenerants was confirmed.
Abstract: Protocol was developed for high frequency in vitro multiplication of an endemic species, Zingiber rubens Roxb. The sprouted buds of the rhizomes were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA; 0.5–5.0 mg dm−3), indole-3-acetic acid (IAA; 0.5–2.0 mg dm−3), kinetin (KIN; 1.0–3.0 mg dm−3), naphthaleneacetic acid (NAA; 0.5–1.0 mg dm−3) and adenine sulphate (ADS; 80–100 mg dm−3). MS basal medium supplemented with 3 mg dm−3 BA and 0.5 mg dm−3 IAA was optimum for shoot elongation. The elongated shoots (1–2 cm) were transferred to multiplication medium containing 2 mg dm−3 BA, 1 mg dm−3 IAA and 100 mg dm−3 ADS. The multiplication rate remained unchanged in subsequent subcultures. Upon ex vitro transfer, 85 % of plants survived. Genetic stability of micropropagated clones were periodically evaluated at an interval of 6 months up to 30 months in culture using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis and genetic uniformity in all regenerants was confirmed.
TL;DR: -1 increased the rooting percentage, number of roots, and root length more efficiently than NAA and higher concentrations of IBA and NAA showed poor results of rooting.
Abstract: -1 increased the rooting percentage (97.5%) number of roots (4.70) and root length (3.47 cm) more efficiently than NAA. Higher concentrations of IBA and NAA (3.0 mg l -1 ) showed poor results of rooting.
TL;DR: It is concluded that kinetin can alleviate the negative impact of salt on sage plants cultivated under arid environments with salinity problems, and seemed to counterbalance or alleviate the stress symptoms induced by salinity.
TL;DR: The results enable the feasible production of CPT of O. alata by means of a cell culture strategy and can help safeguard the plant from extinction.
Abstract: 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.
TL;DR: In this study, an efficient protocol for shoot regeneration from leaves and roots was developed for Centaurea ultreiae, a critically endangered species and connectivity between vascular tissues of regenerated shoots and cambial cells of leaf explants was revealed.
Abstract: In vitro culture is currently used to produce plant material for ex situ conservation of endangered species. In this study, an efficient protocol for shoot regeneration from leaves and roots was developed for Centaurea ultreiae, a critically endangered species. Organogenesis from leaf and root explants was promoted by incubating these explants on half-strength Murashige and Skoog (MS) medium in the presence of one of four different cytokinins [6-benzyladenine (BA), zeatin, kinetin or N6-(2-isopentenyl) adenine (2iP)], each provided at five different levels. Shoot organogenesis was induced in both explants. The best response, 90% of leaf explants producing a mean of 2.48 shoots per explants and 94.3% of root explants producing a mean of 5.60 viable shoots per explants, was observed when explants were incubated on a medium containing 0.55 μM BA. Histological studies revealed connectivity between vascular tissues of regenerated shoots and cambial cells of leaf explants. Moreover, adventitious shoots were derived from pericycle cells of root explants and parenchymatic cells of callus tissues.
TL;DR: Prepared derivatives with certain substitutions of the N9-atom of the purine moiety enhanced the cytokinin activity of the parent compound in the bioassays to a remarkable degree but negatively affected its perception by CRE1/AHK4 and AHK3.
TL;DR: Methods of scratching the seed coat with iron particles and treating the whole seeds with concentrated H2SO4, ultrasonic waves and gamma rays were employed for breaking the seed dormancy, and the effect of gamma irradiation on the growth of caper shoots in vitro was studied.
TL;DR: High growth capacity of cauliflower artificial seeds in commercial substrates is showed which is considered a promising step for their direct use in vivo.
Abstract: An effective protocol for the mass production of cauliflower microshoots was refined using the meristematic layer of cauliflower curd. After the meristematic layer was surface sterilized and shaved off, a commercial blender was used for homogenization and several blending treatments were tested in the range 15–120 s and 30 s was found to be optimal in terms of the amount explants produced and their subsequent growth ability. Explants were cultivated in S23 liquid medium (4.4 g L−1 MS (Murashige and Skoog) and 3% v/w sucrose) supplemented with several combinations of plant growth regulators (PGRs) including 1 and 2 mg L−1 of Kinetin in combination with three types of auxins (indole butyric acid (IBA), Naphthaleneacetic acid (NAA) and Indole-3-acetic acid (IAA)), each at 1 and 2 mg L−1 concentration. The use of 2 mg L−1 Kinetin and 1 mg L−1 IBA gave the best results in terms of its effects on explant induction. Microshoots of different sizes were encapsulated in a sodium alginate matrix and the optimal stage suitable for the production of artificial seeds was assessed in terms of both subsequent conversion and plantlet viability. The feasibility of cultivating cauliflower artificial seeds in commercial substrates (compost, vermiculite, perlite and sand) irrigated with different solution mixtures including sterilized distilled water (SDW), PGRs-free S23 medium and S23 medium supplemented with Kinetin (1 and 2 mg L−1) and IBA or NAA at (1 and 2 mg L−1) was investigated. The use of 2 mg L−1 Kinetin and 2 mg L−1 NAA applied with S23 gave the optimal response with both perlite and compost. This study showed high growth capacity of cauliflower artificial seeds in commercial substrates which is considered a promising step for their direct use in vivo.