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Tomato (Lycopersicon esculentum Mill) belongs to the Solanaceae Lycopersicon widely cultivated throughout the world, only the breeding of new tomato varieties using conventional breeding methods, progress has been slow, and anther culture and free microspore culture technology can get pure line, help to improve the breeding efficiency. At the same time, the tomatoes are self-pollinating crops, heterosis is very obvious and very important Sterile Tomato their crossbreeding. Tomato anther culture free microspore culture influencing factors, from the 'Rigger 87-5' through anther culture 'Stone Fan No. 9', 'thick fur flour' in pollen plants; through free for the first time microspore culture from the 'Rigger 87-5', 'Golden Delicious', 'Diamond' and 'thick fur pink' won a lot of ball-type embryos, the heart-shaped embryo. At the same time, the use of Agrobacterium tumefaciens in genetic transformation of tobacco and tomato total of 25 turn Barnase gene tobacco and 20 to turn Barnase gene tomato, and sterility of biological characteristics were observed in transgenic plants. The specific results are as follows: 1. Tomato anther callus induction and plant regeneration experimental system. In this study, six tomato varieties anther materials research the tomato anther embryogenesis and plant regeneration. Research shows that genotypes are the determinants of tomato anther culture, callus induction was significantly different in different genotypes; potato juice added to the culture medium, activated carbon, six varieties of callus induction rate than the control were reduced 35.14 %, 95.48%; 1.0m / L 2,4-D and 0.5mg / L KT medium supplemented with 'Rigger 87-5' and 'Stone Fan No. 9' induction rate are the highest; culture the added 30g / L sucrose, the 'riegel 87-5' base callus induction rate of 20.24%; medium added 10μmol / L silver nitrate, anther tissue browning rate than the control reduces 70.59%. Tomato anther callus MS ZT1.5mg / L IAA0.2mg / L medium, the induction rate of adventitious buds up to 40%, this recipe is the tomato anther callus differentiate into adventitious buds the better recipe; in adventitious buds 2mg / L the triacontanol differentiation medium, you can improve the quality differentiation rate of adventitious buds, significantly increase the percentage of available bud; adventitious bud to 3-4cm size, access 1/2MS 2.0 mg / LIBA rooting tomato pollen plants. 2. Tomato microspores free cultured embryoid research. Microspore isolation and culture systems, WM and SMC technology used in this experiment two different ways. The results show that, WM technology is the better technology system for the isolation and culture of tomato microspores. The SMC technology than WM than 'Riegel 87-5' and 'Golden Delicious' embryo production was reduced by 53.56% and 49.86%. Different genotypes embryogenesis ability is very different. The experiment three tomato varieties in the same culture conditions, microspore embryo production in descending order: the 'Rigger 87-5', 'Golden Delicious', 'Diamond' 'thick fur pink', '002 '. 'The Rigger 87-5', 'Golden Delicious' and' diamond 'easier to form embryoid '002' embryo production is only 0.13 in the same culture conditions, and difficult to produce embryoids. In tomato microspores free culture process, the immature ovary Add tomato, five varieties of microspore embryo production are significantly higher than the control. Transformed male sterility Barnase gene tobacco, tomato plants get. By the freeze-thaw method into Agrobacterium tumefaciens LB4404, to obtain turn Barnase gene the tobacco and turn Barnase gene tomato. The gene tobacco and turn Barnase of of the turn Barnase gene of tomato by kanamycin screening, PCR, RT-PCR detection showed that the Barnase gene integrated into the genome of tobacco and tomato, and normal transcription and expression in plants. The flower morphology observed filaments to turn Barnase gene tobacco significantly shortened anther flat, no normal pollen grains by tabletting examination. The tobacco not turn Barnase gene seed.
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