2024年10月4日 星期五
6 个烟草杂交组合花药再生苗的培养和DH群体的构建
Culture of regenerated seedlings from anthers and construction of DH populations of six cross combinations of Nicotiana tabacum
2011年 第20卷 第1期 页码[65-68]    下载全文[0.7MB]  
摘要

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。

Abstract

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 doubling rate of different cross combinations after doubling treatment were analyzed. Also, the difference of chloroplast number in stomata guard cells of seedling leaves of haploid and doubled haploid of cross combination DH1 was studied. The results show that there are significant differences in the emergence seedling number per anther, the survival rate of regenerated seedlings after doubling treatment, the survival rate of transplanted seedlings in field and the chromosome doubling rate among different cross combinations. In which, the emergence seedling number per anther of six cross combinations is 1. 57-3. 30, that of cross combination DH5 is the highest with a value of 3. 30 and that significantly higher than other five cross combinations (P<0. 05). After doubling treatment, the survivaisl rate of regenerated seedlings is 21. 12% -33. 42%, that of transplanted seedlings in field is 91. 87% - 98. 86%, those two indexes are the highest in cross combination DH6. And the chromosome doubling rate of six cross combinations is 6. 64% -10. 78%, that is the highest in cross combination DH4. After anther cultured for about 15 d, adding H liquid basic medium can obviously promote anther emergence of cross combination DH1 and DH2. The average of chloroplast number in stomata guard cells of seedling leaves of haploid and doubled haploid of cross combination DH1 is 9. 27 and 17. 46, respectively, the ratio is close to 1︰2 with significant difference. Using anther culture and chromosome doubling treatment, DH populations of six cross combinations could be constructed.

关键词烟草; 杂交组合; DH 群体; 花药培养; 染色体加倍; 秋水仙素
Key wordsNicotiana tabacum L.; cross combination; DH population; anther culture; chromosome doubling; colchicine
作者陈学军1, 彭双玉1,2, 罗建蓉1,2, 杨彦明1, 肖炳光1
所在单位1. 云南省烟草农业科学研究院, 云南Ⅰ溪653100; 2. 云南农业大学, 云南昆明650201
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基金项目中国烟草总公司云南省公司科技项目(06A03;07A04;08A05;2010YN02;2010YN15); 国家烟草专卖局科技项目(110200701023)