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《Xinjiang Agricultural Sciences》 2010-07
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Isolation of Protoplast Shuangya No.5 Flax from Apocynum venetum Linn Protoplast and Somatic Hybridization

JIANG Li1,2,MIAO Shu-kui1,JI Qiao-ling1,ZHU Guo-li1(1.College of Life Science and Technology,Xinjiang University,Urumqi 830046,China;2.Food Engineering Department,Xinjiang Institute of Light Industry Technology,Urumqi 830021,China)  
[Objective]The suitable conditions of dissociate protoplast of Linum usitatissimum L.Shuangya No.5 and Apocynum venetum Linn.,electrofusion and protoplast culture were explored,to lay a foundation for further study of somatic hybridization breeding in the two species.[Method]The two kinds of protoplasts derived from established embryogenic cell suspension lines of Shuangya No.5 and A.venetum were fused with electrofusion and cultured.[Result]The proper penetrant on dissociate protoplast of flax and A.venetum were 5.6% sucrose and 0.7 mol/L mannitol,respectively.The proper combination of enzymes all were 0.5% Cellulase Onozuka R-10+1.0% Cellulase Onozuka RS+0.2% Pectolyase Y-23,the aptimal enzymolysis time were 4 h and 3 h,respectively.The proper subculture time of suspensionembryogenic line of flax was the third day.The survival rate of the two kinds of protoplasts all reached 92.0% and the protoplast yield of flax and A.venetum reached 1.24×106/mL and 22.4×106 /mL under optimum condition,respectively.The optimum electrofusion parameters of two kinds of protoplasts were AC 8 v/cm for 25 s—DC 1 500 v/cm for 60 μs in pulse length,one time—AC 8 v/cm for 9 s,twice repeated,under whose conditions the fusion rate reached 20.5%.[Conclusion]Flax and A.venetum could achieve somatic hybridization through protoplast fusion under definite conditions.
【Fund】: 国家自然科学基金(30660095);; 新疆维吾尔自治区教育厅重点科研项目(XJEDU2006108)
【CateGory Index】: S563
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