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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
Author(s) -
Savita ,
Pati Pratap K.,
Virk Gurdeep S.,
Nagpal Avinash
Publication year - 2017
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12617
Subject(s) - biology , methane sulfonate , cotyledon , callus , rough lemon , shoot , murashige and skoog medium , botany , phytophthora , rutaceae , tissue culture , explant culture , ethyl methanesulfonate , in vitro , horticulture , rootstock , mutant , genetics , gene
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 m m 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 m m EMS for 6‐hr duration showed tolerance (24%) up to 75% CF after 4th selection cycle. While, calli treated with 200 m m 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.