罗汉果性别相关的SCAR标记筛选

Screening of SCAR Markers Linked to Sex Determination in Siraitia grosvenorii(Swingle)C.Jeffrey ex Lu et Z.Y.Zhang

以罗汉果雌雄单株各30份为试材,采用随机引物筛选法,筛选与罗汉果性别相关的RAPD标记,并根据特异片段的测序结果,将RAPD标记转化为特异性和稳定性更好的SCAR标记,以期为罗汉果的性别早期鉴定和分子辅助育种研究提供参考依据。结果表明:在330条随机引物中获得了分别与罗汉果雌性和雄性相关的RAPD引物S2124和S343。根据特异序列测序结果,对雌性特异带S2124-1K和雄性特异带S343-1.4K各自设计并合成了2对SCAR引物,经验证,SCAR引物SC2124-1K-14在62℃和SC2124-1K-8在56℃均可扩增出1 019bp的雌性特异带,SC343-1.4K-12在64℃和SC343-1.4K-4在56℃均可扩增出1 373bp的雄性特异带。这些特异的SCAR标记具有较好的应用价值,可为罗汉果的性别早期鉴定提供分子依据。

Thirty female and male individual plants of S.grosvenorii respectively were used as experimental materials.RAPD markers related to sex determination of S.grosvenorii by random primers were screened,and transformed them into SCAR markers based on the sequencing results of specific bands.The results showed that two markers obtained from 330 RAPD primers,S2124 and S343,which related to female and male of S.grosvenorii respectively.According to the sequencing results of specific fragments,four pairs of SCAR markers were synthesized.By testing,the SCAR markers SC2124-1 K-14 and SC2124-1 K-8 could amplify 1 019 bp female-specific fragment on 62℃and56℃respectively,SC343-1.4 K-12 and SC343-1.4 K-4 could amplify 1 373 bp male-specific fragment on 64℃and 56℃respectively.These SCAR markers had promotional values,and they could provide molecular foundation for gender identification of S.grosvenorii in seedling stage.