| 双亚5号亚麻与罗布麻原生质体解离及体细胞杂交 |
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| 引用本文： | 姜丽,苗书魁,计巧灵,朱国丽.双亚5号亚麻与罗布麻原生质体解离及体细胞杂交[J].新疆农业科学,2010,47(7):1336. |
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| 作者姓名： | 姜丽 苗书魁 计巧灵 朱国丽 |
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| 作者单位： | 新疆大学生命科学与技术学院,乌鲁木齐,830046;新疆轻工职业技术学院食品工程系,乌鲁木齐,830021;新疆大学生命科学与技术学院,乌鲁木齐,830046 |
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| 基金项目： | 国家自然科学基金，新疆维吾尔自治区教育厅重点科研项目 |
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| 摘 要： | 目的]探索双亚5号亚麻与罗布麻原生质体制备、电融合和培养的适宜条件,为进一步深入进行亚麻与罗布麻体细胞杂交育种研究奠定基础.方法]以已建立的双亚5号亚麻和罗布麻胚性细胞悬浮系为材料,制备原生质体并进行电融合,培养.结果]制备亚麻和罗布麻原生质体的最佳渗透剂分别是5.6;蔗糖和0.7 mol/L甘露醇;游离亚麻和罗布麻原生质体的最适酶组合都是0.5; Cellulase Onozuka R-10+1.0; Cellulase Onozuka RS+0.2; Pectolyase Y-23;最佳酶解时间分别是4和3 h;亚麻细胞最佳继代时间是转瓶后的第3 d,在此条件下原生质体存活率均为92.0;,产量分别达到1.24×106和22.4×106个/mL;2种原生质体电融合的最佳参数依次为:AC 8 v/cm,持续25 s-DC 1 500 v/cm,脉冲宽度为60 μs,脉冲次数1次-交流电时间为9 s,重复次数2次.在此条件下融合率达到20.5;.结论]亚麻与罗布麻原生质体在一定的条件下可实现体细胞杂交.
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| 关 键 词： | 亚麻 罗布麻 胚性细胞系 原生质体 电融合 再生愈伤组织 |
| 收稿时间： | 2010-07-25 |
Isolation of Protoplast Shuangya No.5 Flax from Apocynum venetum Linn Protoplast and Somatic Hybridization |
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| JIANG Li,MIAO Shu-kui,JI Qiao-ling,ZHU Guo-li.Isolation of Protoplast Shuangya No.5 Flax from Apocynum venetum Linn Protoplast and Somatic Hybridization[J].Xinjiang Agricultural Sciences,2010,47(7):1336. |
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| Authors: | JIANG Li MIAO Shu-kui JI Qiao-ling ZHU Guo-li |
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| Abstract: | 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. |
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| Keywords: | flax Apocynum venetum Linn embryogenic cell line protoplast electrofusion regenerative callus |
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