红麻雄性不育细胞质相关的cSNP位点发掘

在前期已获得红麻atp9基因编码区的基础上,以红麻细胞质雄性不育系P3A、保持系和F1代为材料,随机选取不同材料的3个atp9独立gDNA克隆子进行测序和序列比对。结果显示:具有不育细胞质的不育系和F1代与具有可育细胞质的保持系在第290位点处存在碱基差异,具有不育细胞质的材料均为碱基T,而具有可育细胞质的材料在相应位点为碱基A;atp9基因第290位点处T/A碱基转换的cSNP位点经已知细胞质的红麻种质资源的进一步检测,同时结合这些资源的田间育性进行分析发现,红麻种质资源UG93-2MS-1、UG93-2MS-2、UG93-2MS-3、UG93-2-22、KN250、KN142、ZB90在第290位点处为碱基T,能扩增出319bp条带,在田间对应的育性除品种UG93-2-22为可育外,其余均为不育或半不育;而福红992在第290位点处为碱基A,未能扩增出319bp条带,在田间对应的育性为可育。因此,位于atp9基因的第290位点处的T/A碱基转换的cSNP位点与红麻不育细胞质相关或连锁,为红麻不育细胞质鉴定提供了新的检测手段。

Development of the molecular marker cSNP associated with male sterile cytoplasm in kenaf

Based on the coding sequences of atp9 obtained in our previous study, three random and separate atp9 gDNA clones of the CMS (Cytoplasmic male sterility) line P3A, its maintainer P3B and F₁ progeny were sequenced and their atp9 sequence alignment were analyzed in kenaf.The results showed that there existed a base differentiation located on 290 th of atp9 between male sterile cytoplasm and male fertile cytoplasm in kenaf.The cytoplasmic male sterility line and F₁ progeny containing the male sterile cytoplasm have base T, however the maintainer line containing male fertile cytoplasm has base A.Some germplasm resources containing known cytoplasm were further analyzed combined with sterility analyses using sequencing and PCR amplification based on the molecular marker cSNP (T/A transformation) located on 290 th of atp9.The results showed that the varieties of UG93-2MS-1, UG93-2MS-2, UG93-2MS-3, UG93-2-22, KN250, KN142, ZB90 have base T and could be amplified 319 bp fragment, and the fertility are sterility or semi-sterility except the variety of UG93-2-22, the variety of Fuhong 992 containing male fertile cytoplasm has base A and had no amplified products.These results suggested that the cSNP (T/A transformation) located on 290 th of atp9 was associated with male sterile cytoplasm or linked to male sterile cytoplasm.It can be developed into new molecular marker to identify sterile cytoplasm.