6个烟草杂交组合花药再生苗的培养和DH群体的构建

以6个烟草(Nicotiana tabacum L.)杂交组合的花药为实验材料,对各杂交组合花药再生苗出苗状况及培养过程中添加H液体基本培养基对花药出苗状况的影响进行了比较分析;在此基础上,采用质量体积分数0.4%秋水仙素浸苗法构建加倍单倍体(DH)群体,对加倍处理后不同杂交组合再生苗的成苗率、大田移栽成活率及染色体加倍率等进行分析,并对杂交组合DH1单倍体和加倍单倍体植株叶片气孔保卫细胞叶绿体数的差异进行了研究。结果表明:不同杂交组合每枚花药的出苗数、加倍处理后再生苗的成苗率、大田移栽成活率及染色体加倍率有较大差异。其中,每枚花药出苗数为1.57～3.30,杂交组合DH5每枚花药的出苗数最多,达到3.30株,显著高于其他5个杂交组合(P<0.05);加倍处理后再生苗的成苗率为21.12%～33.42%,大田移栽成活率为91.87%～98.86%,杂交组合DH6的成苗率和大田移栽成活率最高;染色体加倍率为6.64%～10.78%,杂交组合DH4的染色体加倍率最高。花药培养约15 d后,添加H液体基本培养基能够显著促进杂交组合DH1和DH2花药出苗。杂交组合DH1单倍体苗和加倍单倍体苗叶片气孔保卫细胞的平均叶绿体数分别为9.27和17.46,二者比值接近1∶2,差异显著。通过花药培养和染色体加倍处理,分别获得了6个烟草杂交组合的DH群体。

Culture of regenerated seedlings from anthers and construction of DH populations of six cross combinations of Nicotiana tabacum

Taking anthers of six cross combinations of Nicotiana tabacum L.as experimental materials,the emergence status of regenerated seedlings from anthers of different cross combinations and the effect of adding H liquid basic medium in culture process on emergence status were comparatively analyzed.On this basis,doubled haploid(DH) populations were constructed by soaking seedling method with 0.4% colchicine,and the survival rates of regenerated seedlings and transplanted seedlings in field and the chromosome dou...