In vitro mutagenesis and characterization of mutants through morphological and genetic analysis in orchid Aerides crispa Lindl.
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Date
2018-06
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NISCAIR-CSIR, India
Abstract
Mutation breeding techniques combined with tissue culture and molecular marker methods provide a powerful tool for improvement of slow growing plants such as orchids. In this study, we developed a protocol for in vitro mutagenic studies in a medicinal orchid of Western Ghats, Aerides crispa Lindl., commonly called the Curled Aerides. In vitro grown 60 day old protocorms treated with low concentrations (0.025–0.03%) of colchicine, ethyl methane sulphonate (EMS), and low doses (1–4Gy) of gamma radiations, and 27 combinations resulted in healthy multiple shoot bud formation with 40–60% survival frequency. Higher singly or combined dose treatments caused protocorm or shoot initials death, after 20–30 days. Healthy 2500 in vitro seedlings were screened for phenotypic changes in shoots and roots owing to a mutagenic effect. In total, 206 in vitro seedlings of 52 variant lines were identified on the basis of their unique leaf shape, colour, white stripes, thickness, length, and width and their root length and thickness. These variant lines were multiplied, established in an orchidarium, and compared with the control for genetic variability by using Random Amplified Polymorphic DNA method. Only 15 genetically distinct mutant lines were identified, which exhibited disparity in growth rate, leaf shape, leaf length, width, chlorophyll variegation, stomatal density, and/or pigment contents.
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Srivastava Deepti, Gayatri MC, Sarangi SK. Molecular characterization of bacterial biocontrol agents and their chitinase genes from tea soil. Indian Journal of Experimental Biology. 2018 Jun; 56(6): 395-401