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    曹景林, 王欣, 程君奇, 李亚培, 吴成林, 张俊杰, 蔡长春. 烟草未授粉子房离体培养诱导胚状体的影响因素研究[J]. 中国烟草科学, 2023, 44(6): 84-90. DOI: 10.13496/j.issn.1007-5119.2023.06.012
    引用本文: 曹景林, 王欣, 程君奇, 李亚培, 吴成林, 张俊杰, 蔡长春. 烟草未授粉子房离体培养诱导胚状体的影响因素研究[J]. 中国烟草科学, 2023, 44(6): 84-90. DOI: 10.13496/j.issn.1007-5119.2023.06.012
    CAO Jinglin, WANG Xin, CHENG Junqi, LI Yapei, WU Chenglin, ZHANG Junjie, CAI Changchun. Study on the Factors Influencing the Induction of Embryoids by in vitro Culture of Unpollinated Ovary in Tobacco[J]. CHINESE TOBACCO SCIENCE, 2023, 44(6): 84-90. DOI: 10.13496/j.issn.1007-5119.2023.06.012
    Citation: CAO Jinglin, WANG Xin, CHENG Junqi, LI Yapei, WU Chenglin, ZHANG Junjie, CAI Changchun. Study on the Factors Influencing the Induction of Embryoids by in vitro Culture of Unpollinated Ovary in Tobacco[J]. CHINESE TOBACCO SCIENCE, 2023, 44(6): 84-90. DOI: 10.13496/j.issn.1007-5119.2023.06.012

    烟草未授粉子房离体培养诱导胚状体的影响因素研究

    Study on the Factors Influencing the Induction of Embryoids by in vitro Culture of Unpollinated Ovary in Tobacco

    • 摘要: 源于未授粉子房的雌性单倍体在烟草育种利用价值上优于源于花药或小孢子的雄性单倍体,开展烟草未授粉子房的离体培养对烟草单倍体育种具有重要意义。为了建立未授粉子房离体培养技术体系,分别以不同的杂种F1代为试材,采用HW培养基探讨了消毒方法、接种方式、接种密度、无机盐浓度、外源激素配比和琼脂浓度6个关键因素对胚状体诱导的影响。结果表明:花蕾不需剥开花瓣而直接放入0.1%升汞中浸泡8 min,剥开子房壁并横切子房后接种,每个125 mL三角瓶中接种2块子房,培养基中无机盐浓度额外增加60%、附加IAA 0.5 mg/L和6-BA 2 mg/L或者附加KT 2.0 mg/L和2,4-D 0.5 mg/L、琼脂浓度9 g/L有助于对胚状体的诱导。据此进一步完善了烟草未授粉子房离体培养过程中的技术参数。

       

      Abstract: The utilization value of female haploids derived from unpollinated ovaries is superior to male haploids derived from anthers or microspores in tobacco breeding. Therefore, in vitro culture of unpollinated ovary is of great significance for tobacco haploid breeding. In order to establish the technique system for in vitro culture of unpollinated ovaries, different F1 hybrids were used as test materials to explore the effects of six key factors, including disinfection method, inoculation method, inoculation density, inorganic salt concentration, exogenous hormone ratio, and agar concentration, on embryoid induction in HW media. The results showed that the measures that contributed to the induction of embryoids were as follows:the buds were immersed in 0.1% HG solution for 8 min without peeling the petals, the ovary walls were peeled off and the ovaries were transected before inoculation, 2 ovaries were inoculated in each 125 mL triangular flask, and the inorganic salt concentration in HW media was increased by an additional 60%, with additional IAA 0.5 mg/L and 6-BA 2 mg/L, or KT 2.0 mg/L and 2, 4-D 0.5 mg/L, and agar concentration of 9 g/L. Based on this, the technical parameters for in vitro culture of unpollinated tobacco ovaries were further improved.

       

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