烟草雄性不育未受精胚珠离体培养和植株再生  

In vitro culture and plant regeneration of unfertilized male sterile ovules of tobacco

作　　者：曹景林 程君奇 李亚培 吴成林 王欣 CAO Jinglin;CHENG Junqi;LI Yapei;WU Chenglin;WANG Xin(Tobacco Research Institute of Hubei Province,Wuhan 430030,China)

机构地区：[1]湖北省烟草科学研究院,武汉市430030

出　　处：《烟草科技》2023年第5期1-7,共7页Tobacco Science & Technology

基　　金：中国烟草总公司湖北省公司科技重点项目“兼抗青枯病和赤星病优质烤烟新品种的分子选育与示范”(027Y2021-007)。

摘　　要：为了探讨烟草未受精胚珠单倍体育种技术,以烟草雄性不育杂交种KRK26为材料,选取5种发育时期的胚珠,在基于H、HW和N_(6)培养基而优化的CM1、CM2、CM3、CM4和CM55种培养基上培养,研究了未受精胚珠离体培养诱导单倍体技术。结果表明,在试验范围内胚囊发育越成熟,胚珠的增殖诱导效果越差,应选取发育较早的胚珠(即花朵的花冠即将露出花萼至花冠比花萼长1倍时期的胚珠)进行培养。在5种优化培养基中,以培养基CM1对胚状体的诱导效果最好,在CM1基础上将琼脂浓度调整到10 g/L利于抑制胚状体玻璃化;将胚状体转移到不含激素的H培养基上培养,能够避免胚状体褐化,利于再生芽诱导;将再生芽转移到加有IAA 10 mg/L的H培养基或N_(6)培养基上培养,利于再生芽快速生根,获得大量的再生植株。流式细胞仪检测发现大部分再生植株为单倍体植株。据此,建立了从雄性不育烟草未受精胚珠诱导单倍体植株的培养体系。In order to investigate the haploid breeding technique for unfertilized ovules of tobacco,using tobacco male sterile hybrid KRK26 as the test material,five types of ovules at development stage were selected and cultured on CM1,CM2,CM3,CM4 and CM5 media which were optimized based on H,HW and N_(6)media.The in vitro haploid induction technique of unfertilized ovules was studied.The results showed that the more mature the embryo sac was,the worse the effect of ovule proliferation induction would be.The ovules developed earlier,i.e.from the time when the flower’s corolla was about to expose calyx to the time when the corolla was twice as long as the calyx,should be selected for culture.Among the five optimized media,medium CM1 had the best induction effect on the embryoid.Adjusting the agar concentration to 10 g/L on the basis of CM1 was beneficial to the inhibition of the embryoid vitrification.Transferring the embryoids to medium H without hormone could avoid the browning of the embryoids and was beneficial for the induction of regenerated buds.Transferring the regenerated buds to medium H or N_(6)with IAA 10 mg/L for the culture was helpful for the rapid rooting of the regenerated buds and a large number of regenerated plants could be obtained.Flow cytometry results indicated that most of the regenerated plants were haploids.These results demonstrated that the culture system for inducing haploid plants from unfertilized ovules of male sterile tobacco was established.

关 键 词：烟草 雄性不育 未受精胚珠 单倍体 再生植株 

分 类 号：S572.01[农业科学—烟草工业]