| 2 pollinations between seedless grapes( Dalihong Seedless X Sultanina, Ruby Seedless XOlmo Seedless), 11 pollinations between seedless grapes and Chinese wild grapes (Flame SeedlessXHeilongjiang Seedling,Olmo SeedlessXZhuoshan-l,Changsui Seedless X Beichun, Sultanina X Yingyu, Ruby Seedless X Red Globe, Sultanina X Zhuoshan-1, Ruby SeedlessX Beichun, Dalihong Seedless XHe'an, Zhenzou Seedless X Heilongjiang Seedling, Jingzaojin Seedless X Heilongjiang Seedling, Jingzaojin Seedless X Yingyu), open-pollinated Dawn Seedless and self-pollinated Sultanina were made to investigate effect of harvest time, basic medium for embryo rescue, medium for embryo culture, medium for embryo development, medium for embryo germination, medium for embryo rescue plant development, conditions of young plant tissue culture for propagation and transplanting. Anatomy and morphology of embryos were investigated in succession with materials of open-pollinated Dawn Seedless, open-pollinated Beichun(seeded hybrid) and grapes of Flame Seedless pollinated with Heilongjiang Seedling. Enzyme activities of POD, CAT and SOD, soluble starch(SS), soluble protein(SP) and soluble sugar(S) in embryos were determined during the embryo development of seedless grapes, seeded grapes.The main results were as follows.1.By embryo rescue, 87 plants were obtained from 13 crosses, including 17 plants from hybrid Dalihong Seedless X Sultanina, 6 plants from hybrid Sultanina X Yingyu, 12 plants from hybrid Flame Seedless X Heilongjiang Seedling, 11 plants from hybrid Changsui Seedless X Beichun, 10 plants from hybrid Olmo Seedless X Zhuoshan-1, 18 plants from open-pollinated Dawn Seedless and 13 plants from self-pollinated Sultanina.2.The suitable basic media were selected for embryo development, embryo germination and embryo rescue plant development. 40d after pollination, ovules of open-pollinated Dawn Seedless and open-pollinated SP137 Seedless were cultured in vitro on basic mediums of 65, MS and Nitsch and it showed that the best medium for embryo rescue was Nitsch Medium.It was found that basic medium with the addition of 6% sucrose, 0.1% PVP and 500mg·L-1 casein hydrody was best for embryo development; 3% sucrose, 0.1% PVP and 300mg·L-1 casein hydrody was best for embryo germination. 10-20 days after embryo germination, young plants were transferred to grow on 1/2MS with 2% sucrose.inFor embryo development, basic medium with the addition of 2.5mg -I/-1BA+O-Smg -I/-GAs+O-Smg -1/-0.1mg -L-1 6-BA +ZT was suitable for 6 crosses including Flame Seedless X Heilongjiang Seedling, SultaninaX Zhuoshan-1, Ruby Seedless X Beichun, Dalihong Seedless X He-an, Jingzaojin Seedless X Heilongjiang Seedling, Jingzaojin Seedless X Yingyu; 2.0mg·L--lBA+0.5mg·L~-GA3 +0.5mg·L-!6-BA + 0.1 mg·L-1 ZTwas suitable for 7 crosses including Ruby Seedless X Red Globe, Changsui Seedless X Beichun, Dalihong Seedless XSultanina, Olmo Seedless X Zhuoshan-1, Ruby Seedless X Olmo Seedless, Zhenzou Seedless X Heilongjiang Seedling, open-pollinated Dawn Seedless and self-pollinated Sultanina;For embryo germination, basic medium with the addition of l.Smg·L/-1BA+1.0mg·L--6-BA +0.2mg·L/-GA3 was suitable for 6 crosses including Changsui Seedless X Beichun, Sultanina X Taishan-1, Dalihong Seedless X Sultanina, open-pollinated Dawn Seedless, Olmo Seedless X Zhuoshan-1, Zhenzhou Seedless X Heilongjinag Seedling; 1.0mg·I/-1BA +1.0 mg·L-16 BA +0.2 mg·L-1GA for Flame Seedless X Helongjiang Seedling, self-pollinated Sultanina, Ruby Seedless X Beichun, Ruby Seedless X Olmo Seedless, Jingzaojin Seedless X Heilongjiang Seedling, Jingzaojin Seedless X Yingyu.For establishment of embryo rescue plants, basic medium with the addition of O.lmg ·L-1IBA+0.02mg ·L--6-BA was suitable for Flame SeedlessX Helongjiang Seedling, Changsui Seedless X Beichun, Dalihong Seedless X Sultanina, self-pollinated Sultanina, open-pollinated Dawn Seedless, SultaninaX Yingyu, Olmo Seedless X Zhuoshan-1.For tissue culture of embryo rescue plants, 1/2MS... |
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