摘要
为得到一种鉴别糯质谷子的分子标记方法,用于优质谷子糯质资源的利用及糯质品种的选育。通过对糯性谷子十里香与梗性谷子豫谷1号配制的F2群体的F3种子胚乳进行碘液检测,结果显示:深蓝色与棕红色胚乳单株数的比例符合3∶1,说明糯质对梗质是由单基因控制的隐性性状。利用糯质分型引物对十里香基因组进行PCR扩增和产物测序,结果表明,十里香的糯性是由Ⅳ型糯质基因控制,即由TSI-2转座子插入到淀粉合成酶基因(Si006103m)内元1形成。根据该糯质基因设计出2对In Del标记引物waxy-TSI2/int1和waxy-int1-1F/4R,扩增谷子基因组DNA,若waxy-TSI2/int1引物组合能扩增出984 bp的扩增片段,并且waxy-int1-1F/4R引物组合不能扩增出540 bp的扩增片段,则该材料为含Ⅳ型糯质基因的糯质谷子。最后,利用该标记组合从100份谷子资源中鉴定到2份属于Ⅳ型糯质基因控制的糯质材料。可见,该标记组合可以准确鉴别谷子Ⅳ型糯质基因。
The study provided a molecular marker technology to identify the waxy foxtail millet,which could be used to utilize the excellent waxy resources of foxtail millet and to breed the waxy varieties. Firstly,through the iodine solution detection of the F3 seeds endosperm from F2 constructed from the hybridization of waxy foxtail millet,Shilixiang,and non-waxy foxtail millet,Yugu 1. Results showed that the ratio of plant numbers with dark blue and brownish red endosperm was 3 ∶ 1,indicated that the waxy phenotype was a recessive trait controlled by a single gene. Then using waxy genotyping primers,the genomic DNA of Shilixiang was amplified by PCR and the products were sequenced. The results showed that the waxy trait was controlled by a Ⅳ type waxy gene,namely a TSI-2 transposon was inserted into the first intron of the starch synthase gene（ Si006103m）. Therefore,the In Del molecular markers,waxy-TSI2/int1 and waxy-int1-1F/4R,were developed based on the gene sequence of Si006103 m,and then the genomic DNA of foxtail millet was amplified by them. If there was an product with 984 bp amplified by the waxy-TSI2/int1 primer and no product with 540 bp amplified by the waxy-int1-1F/4R primer,the material was identified as a waxy foxtail millet containing the type Ⅳ waxy gene. Furthermore,we identified 2 waxy materials controlled by the Ⅳ type waxy gene from 100 foxtail millet resources using two pairs of waxy primers. Therefore,the InDel markers can be used to accurately identify the Ⅳ type waxy gene from foxtail millet.
作者
白辉
李志勇
王永芳
石灿
刘磊
全建章
董志平
BAI Hui LI Zhiyong WANG Yongfang SHI Can LIU Lei QUAN Jianzhang DONG Zhiping(Institute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences,National Foxtail Millet Improvement Center, Minor Cereal Crops Laboratory of Hebei Province, Shijiazhuang 050035, Chin)
出处
《华北农学报》
CSCD
北大核心
2016年第4期7-12,共6页
Acta Agriculturae Boreali-Sinica
基金
国家现代谷子糜子产业技术体系建设项目(CARS-07-12.5-A8)
科技部“十二五”农村领域国家科技计划课题(2011BAD06B01-1)
关键词
谷子
糯质基因
分子标记
共分离
Foxtail millet
waxy gene
Molecular marker
Co-segregated
