Objective: In vitro shoot regeneration of Decalepis hamiltonii Wight and Arn. is an endangered endemic medicinal plant using biotechnological interventions and to conserve this threatened species. Methods: In the present study, various explants such as shoot tip, leaf, and nodal segments were inoculated on Murashige and Skoog media augmented with different hormonal regimes of auxin and cytokinin combinations, namely, naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), benzyl adenine (BAP), 6-(γ,γ-Dimethylallylamino)purine (2iP), and triacontanol (TRIA). Results: Direct regeneration of shoots obtained in 3.0 mg/l 2iP alone and in combination with 0.1 mg/l IAA and/or 1.0 mg/l BAP exhibited the best response with average shootlet length being 6.5±0.17–8.0±0.92 cm, respectively, and percentage response was between 68% and 75%. The callus induced regeneration was obtained from both nodal and leaf explants with maximum response (85%) observed in combination of (2.0 mg/l) 2iP, (1.0 mg/l) IAA and (2.0 mg/l) kinetin with multiple shoots showing mean shoot number of 1.83 and average shootlet length of 6.3±0.19 cm. Conclusions: The current research provides a competent in vitro propagation method for Decalepis which could be commercialized for developing identical plants with good mass multiplication rate and for better conservation of the germplasm.
Basella spp. a perennial vine of Basellaceae family used as a leafy vegetable. Phytonutrient of Basella spp. is being exploited in Indian medicinal system since antiquity for its antifungal, anticonvulsant, antiin ammatory, antipyretic, antiulcer and analgesic properties. Propagation of Basella through seeds have limitations for germination and owering, in vitro regeneration was studied on MS (Murashige& Skoog) media supplemented with various plant growth regulators, indole acetic acid (IAA), Indole butyric acid (IBA), 1-naphthalene acetic acid (NAA), N,6-benzyladenine (BA), kinetin (KIN), zeatin (ZEA), gibberillic acid (GA 3 ), and adenine sulphate (ADS), silver nitrate (AgNO 3 ) as additives. The shoot bud initiation was observed in all the combinations studied showing a good response for direct regeneration. Shooting (84%) was observed in 12 days after inoculation in 1mg l − 1 BA + 0.1 mg l − 1 NAA. Liquid media containing 0.1mg l − 1 BA + 0.5 mg l − 1 KIN + 0.1 mg l − 1 IAA was preeminent in multiple shoots (22 ± 0.13) production with average shoot length (5.81 ± 0.19) in 5 weeks (wk). Supplementation of 40 mg l − 1 AgNO 3 and 40 mg l − 1 ADS to media containing 1mg l − 1 BA + 0.1 mg l − 1 NAA resulted in enhanced number of elongated shoots with number of leaves. In vitro owering was obtained on MS media containing 0.5mg l − 1 BA + 0.5 mg l − 1 GA 3 concentrations. The survival rate of hardened plants was 90% after transferring to soil. This protocol can be e ciently used for mass production for regeneration of genetically transformed Basella spp. in studying its metabolite pro le specially betalains and transformation of Basella.
Since ages, plants continue to provide new remedies to mankind. Hemidesmus indicus L. R. Br. is one such plant belonging to family Apocynaceae, showing potent medicinal properties known through traditional knowledge. Hemidesmus is also explored for the presence of flavoring compound namely 2-hydroxy-4-methoxybenzaldehyde (HMB) which is used in pharmaceutical and nutraceutical industries. Due to anthropogenic activities, the plant has been exploited till the ridge for its ethnobotanical properties for mankind. Biotechnological intervention to conserve this endangered sps through in vitro plant cultures, micropropogation and genetic transformation studies is the pre-requite to maintain it from extinction. The objective of the study is to improve the regeneration potential and optimize the genetic transformation in Hemidesmus indicus. The direct regeneration of Hemidesmus indicus through leaf explants, nodal explants with subsequent plant regeneration using Murashige and Skoog (MS) medium supplemented with various plant growth regulators (auxins, cytokinins, and gibberellic acid), adenine sulphate, TRIA. The Agrobacterium tumefaciens mediated genetic transformation studies in Hemidesmus indicus was carried out in callus cultures using the plant expression vector pCAMBIA 1301. The caulogenic response of 78.8%, 73.3% and 71.4% was observed when the leaf explant was inoculated on MS media containing 2.3 mgL− 1 BAP + 0.2 mgL− 1 2,4-D, 0.02 mgL− 1 TRIA + 2 mgL− 1 BAP, 1 mgL− 1 KIN + 1 mgL− 1 NAA respectively with creamish yellow nodular friable callus by the 4 weeks. The initiation of shoot bud was observed within three days after inoculation of nodal explant on media supplemented with 1 mgL− 1 BAP + 0.1 mgL− 1 NAA, 1 mgL− 1 BAP + 0.1 mgL− 1 NAA + 40 mgL− 1 AgNO3, 1 mgL− 1 BAP + 0.1 mgL− 1 NAA + 40 mgL− 1 AgNO3 + 40 mgL− 1 adenine sulphate respectively and incubated in the dark for 2 weeks. Shoot regeneration from the leaf explants was also observed within 4 weeks after inoculation in MS medium with 1 mgL− 1 BAP + 0.1 mgL− 1 NAA. Agrobacterium mediated genetic transformation was carried out successfully in callus cultures of H. indicus. The transformation efficiency was found to be 26%. The efficient shoot regeneration was observed within 4 weeks and transformation study can be further applied for over expression of biosynthetic genes to enhance the bioactive components that have immense significance in pharmaceutical, nutraceutical and cosmetic industries.
Basella spp. a perennial vine of Basellaceae family used as a leafy vegetable. Phytonutrient of Basella spp. is being exploited in Indian medicinal system since antiquity for its antifungal, anticonvulsant, anti-inflammatory, antipyretic, antiulcer and analgesic properties. Propagation of Basella through seeds have limitations for germination and flowering, in vitro regeneration was studied on MS (Murashige& Skoog) media supplemented with various plant growth regulators, indole acetic acid (IAA), Indole butyric acid (IBA), 1-naphthalene acetic acid (NAA), N,6-benzyladenine (BA), kinetin (KIN), zeatin (ZEA), gibberillic acid (GA3), and adenine sulphate (ADS), silver nitrate (AgNO3) as additives. The shoot bud initiation was observed in all the combinations studied showing a good response for direct regeneration. Shooting (84%) was observed in 12 days after inoculation in 1mg l− 1 BA + 0.1 mg l− 1 NAA. Liquid media containing 0.1mg l− 1 BA + 0.5 mg l− 1 KIN + 0.1 mg l− 1 IAA was preeminent in multiple shoots (22 ± 0.13) production with average shoot length (5.81 ± 0.19) in 5 weeks (wk). Supplementation of 40 mg l− 1 AgNO3 and 40 mg l− 1 ADS to media containing 1mg l− 1 BA + 0.1 mg l− 1 NAA resulted in enhanced number of elongated shoots with number of leaves. In vitro flowering was obtained on MS media containing 0.5mg l− 1 BA + 0.5 mg l− 1 GA3 concentrations. The survival rate of hardened plants was 90% after transferring to soil. This protocol can be efficiently used for mass production for regeneration of genetically transformed Basella spp. in studying its metabolite profile specially betalains and transformation of Basella.
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