DEVELOPMENT OF SCAR MARKERS FOR RESISTANCE TO FUNGAL DISEASES FROM CRAB APPLE BIOTYPES
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Date
2019-05
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UHF,NAUNI
Abstract
ABSTRACT
To facilitate the use of marker assisted selection in apple breeding programme, six
PCR based specific sequence characterized amplified region (SCAR) markers were
developed from crab apple. Seven biotypes of indigenous crab apple biotypes maintained at
two field gene banks of Himachal Pradesh state of India were used to identify RAPD markers
for resistance to apple scab and woolly aphis. RAPD molecular analysis was evaluated
among seven biotypes using 119 primers. Among these, 94 primers generated polymorphism.
Four primers OPB-12, OPB-14, OPD-06 and OPA-18 amplified five unique bands of 500bp,
1kbp, 1kbp, 1kbp and 1kbp in M. baccata Shillong, M. baccata Khrot, M. baccata Kinnaur
(Dhack), M. baccata Kashmir and M. baccata J&K respectively. These markers were gel
purified and cloned into easy cloning TA vectors. Plasmid of the confirmed positive clones
after restriction digestion and colony PCR was subjected to sequencing and homology search.
Sequences showing homology with the apple scab and woolly aphid resistant genes were
used to design pairs of six SCAR primers, which specifically amplified these RAPD
fragments in crab apple biotypes as well as M. floribunda and apple rootstock MM106 of
known resistance. No amplification was observed in commercially grown varieties of apple,
therefore, it is verified that all six SCAR markers were able to distinguish between the
resistant and susceptible apple genotypes. Molecular marker assessment with already
developed nine SCAR markers in fourteen individual plants of crab apple verified their
presence/absence and in accordance with the bioassay results.
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