In vitro selection of resistant/tolerant mutants lines of Citrus jambhiri Lush. using crude culture filtrate of Phytophthora parasitica and their randomly amplified polymorphic DNA analysis

Abstract

This study deals with in vitro-induced mutagenesis and selection of Phytophthora tolerant lines of Citrus jambhiri and their regeneration. For in vitro-induced mutagenesis, cotyledons were treated with ethyl methane sulfonate (EMS) 100, 200 and 300 mm for different time durations viz. 1, 3, 6 and 9 hr. Callus cultures were established from EMS treated cotyledon explants on MS medium supplemented with 2.0 mg/L of 2,4-D and 500 mg/L of malt extract. Calli derived from cotyledon were challenged in vitro on selective MS medium containing 5%-100% of culture filtrate (CF) of the Phytophthora parasitica. Selected mutagen-treated calli showed resistance in vitro on media containing CF. Calli treated with 100 mm EMS for 6-hr duration showed tolerance (24%) up to 75% CF after 4th selection cycle. While, calli treated with 200 mm for 6-hr duration showed maximum tolerance (76%) up to 75% CF. Resistant calli were then transferred to MS regeneration medium for shoot bud regeneration. A dose-dependent decrease in the regeneration capacity of the selected calli was observed with the increasing concentration of the CF. In randomly amplified polymorphic DNA analysis, plantlets showed different banding pattern in comparison with the control plant, which confirms the presence of variations at genetic level. K E Y W O R D S Citrus jambhiri, culture filtrate, ethyl methane sulfonate, Phytophthora parasitica, resistance How to cite this article: Savita, Pati PK, Virk GS, Nagpal A. In vitro selection of resistant/tolerant mutants lines of Citrus jambhiri Lush. using crude culture filtrate of Phytophthora parasitica and their randomly amplified polymorphic DNA analysis. J Phytopathol. 2017;165:771-781.

EMS induced in vitro mutagenesis for solid mutant induction and validation of mutants using morphological and molecular tools in kinnow mandarin (Citrus nobilis × Citrus deliciosa)

“…The in-vitro mutagenic effect of EMS has been tested only in Citrus jambhiri (Lush.) and C. sinensis (L.) Osbeck (Ge et al 2015, Savita et al 2017) however, with a poor plantlet recovery. Further no attempt has so far been made on EMS induced in vitro mutagenesis for solid mutant induction in mandarin which also requires reliable and reproducible cellular totipotency mediated high frequency plantlet recovery techniques.…”

Sequence-related Amplified Polymorphism and Target Region Amplification Polymorphism Markers-based Profiling of Sodium Azide and Ethyl Methanesulfonate-derived Black Rot-resistant Dendrobium sp. ‘Earsakul’ Mutants from In Vitro Mutagenesis and Selection

“…Mutation breeding is an alternative for breeders to develop new cultivars through induction either of radiation or of chemical mutagens because they generate genetic variations or even novel alleles, which may be associated with desirable traits (i.e., disease resistance). Sodium azide (NaN 3 ) and ethyl methanesulfonate (EMS) are popular chemical mutagens that are available for purchase and highly effective (Arisha et al 2014;Kumar et al 2011;Savita et al 2017;Yoosumran et al 2018), and they can be successfully used to induce disease resistance in economically important crops (Chen et al 2014;Lo et al 2022). Generally, both mutagens induce mutations by base substitution, leading to incorrect base-pairing, and they also induce chromosomal abnormalities.…”

“…An array of molecular marker systems, especially arbitrarily amplified DNA markers, i.e., random amplified polymorphic DNA and intersimple sequence repeat (ISSR), can be commonly used to detect and characterize mutant and somaclonal variants (Bello-Bello et al 2014;Savita et al 2017). For target-specific breeding purposes, markers designed to uncover genetic variability should be involved with functional genes because they may reflect functional polymorphisms (Andersen and L€ ubberstedt 2003).…”