DEVELOPMENT OF SEEDLESS GRAPE HYBRIDS THROUGH EMBRYO RESCUE AND CONFIRMATION OF HYBRIDITY BY MOLECULAR MARKERS
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Date
2008-08-08
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University of Agricultural Sciences GKVK, Bangalore
Abstract
A study on development of seedless grape hybrids through embryo
rescue and confirmation of hybridity by molecular markers was carried out with
the objective of rescuing embryos in crosses involving ‘Thompson seedless’ as
the female parent and four downy mildew resistant lines (SV12309, SV23501,
SV12364, SV18315) as the pollen parents, to identify hybrids resistant to
downy mildew (fungal) pathogen Plasmopara viticola, and, to develop molecular
markers for grape hybrids so-generated. Using conventional breeding
procedures, it is not possible to obtain hybrid progeny in grape crosses
involving seedless (‘stenospermocarpic’) female parents. Such crosses can,
however, be salvaged using sequential embryo rescue and hybrids can be
obtained. For culturing ovules, berries of various crosses were collected at 8, 9,
10 and 11 Weeks Post Pollination (WPP). Initiation of growth and maintenance
of ovules was done on Emershad and Ramming semi-solid medium
supplemented with Casein hydrosylate (50mg/l)+ Myo-inositol (50mg/l)+ LCysteine
(211.6 mg/l)+ Morpholino Ethane Sulphonic acid (1000mg/l). At 8
weeks from in vitro incubation, embryos were excised from ovules and subcultured
on semi-solid Woody Plant Medium (WPM) supplemented with benzyl
adenine (BA) @ 0.25mg/l. In four to eight weeks, the embryos started
germinating and subsequently developed into full-fledged plantlets in liquid
medium. Hybrid plantlets were then transferred to polybags containing Soilrite (
75% Peat moss + 25% Perlite). A total of 122 hybrid plantlets were recovered
from 955 embryos sub-cultured (which were excised from 1698 cultured
ovules). The true hybrid nature of these hybrids was confirmed by using ISSR
(UBC-807, UBC-817, UBC-825, UBC-828) markers.
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