Factors affecting in vitro propagation were evaluated for Ceropegia attenuata Hook., an endemic and endangered plant having ornamental potential but a limited reproductive capacity. Rapid shoot multiplication from nodal explants was established using varying concentrations of cytokinins and auxins either alone or in combinations. The highest frequency of shoot induction was achieved when nodal explants were inoculated on Murashige and Skoog (MS) medium supplemented with 13.31 μM 6-benzylaminopurine with a mean of 12.9±0.5 shoots per explant. High concentrations of TDZ (6.81-11.35 μM) and KN (6.78-11.61 μM) resulted in stunted and vitrified shoots. Factors implicated in the promotion of floral transition of the C. attenuata have been identified which are 4-amino-3, 5, 6-trichloropicolinic acid (picloram), 6-benzylaminopurine, sucrose and photoperiod. The highest frequency of flowering (100%) was obtained when axillary shoot explants were transferred to MS medium supplemented with picloram (4.14 μM) within 4 weeks of culture. Transfer of in vitro regenerated shoots to half strength MS medium with 2.46 μM indole-3-butyric acid (IBA) showed maximum root induction. The in vitro grown plantlets were successfully acclimatized in the glasshouse with 85% of survival and showed normal development. The developed protocol provided a simple, cost-effective approach for the conservation of endangered plant C. attenuata for replenishing its declining populations.
The study aimed to develop an efficient, rapid, and large-scale in vitro regeneration system for propagation, conservation, and restoration of an endemic and critically endangered herb, Ceropegia mohanramii. The cultures were established using nodal explants on Murashige and Skoog’s (MS) medium supplemented with 6-benzylaminopurine (BAP: 1.0 mg/l). Nodal buds cultured on MS medium supplemented with BAP (2.0 mg/l) along with indole-3-butyric acid (IBA, 0.5 mg/l) resulted with production of maximum number of shoots (17.1 ± 1.2) in hundred percent of the cultures. MS medium supplemented with BAP (2.0 mg/l) along with diverse concentrations of indole-3acetic acid (IAA) promoted the in vitro flowering. In vitro regenerated shoots were transferred to one-half MS medium fortified with singular supplementation of auxins, where IBA (1.5 mg/l) served optimal for production of maximum number of roots (5.7 ± 0.6). In vitro derived plantlets were hardened under controlled conditions in a glasshouse and subsequently transferred to soil. Over 1200 saplings were transplanted to eight different localities of the Western Ghats where over 76% survival is recorded after 1 year of transplantation.
The genus Chlorophytum consists of medicinally important species like Chlorophytum borivilianum, C. tuberosum and C. attenuatum. Uncontrolled harvest of this plant from wild habitat due to its high commercial value made the species of this genus be listed in the Red Data Book of Indian plants as an endangered species. In India, approximately nineteen species of Chlorophytum are found; out of these, only C. borivilianum is cultivated commercially. The objective of this study was to measure genetic diversity, population structure and phylogenetic relationship among the species using Amplified Fragment Length Polymorphisms (AFLP). Fifteen pairs of primer (out of 64 primer pairs screened) were used to analyse the genetic diversity in eighteen species of genus Chlorophytum. Cluster analysis, estimation of the gene flow among the species and of the phylogeographic distribution of this genus were carried out using an AFLP data matrix. A high level of genetic diversity was observed on the basis of the percentage of polymorphic bands (99.91%), Shannon's information index (0.3592) and Nei's gene diversity (0.2085) at species level. Cluster analysis of UPGMA dendrogram, principal component analysis and Bayesian method analysis resolved these species in three different clusters, which was supported by morphological information. The Mantel test (r=0.4432) revealed a significant positive correlation between genetic and geographic distances. The collected data have an important implication in the identification, authentication, and conservation of the species of the genus Chlorophytum.
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