甘蓝型油菜均隆油5号特征指纹图谱构建和杂种纯度鉴定

Hybrid Purity Identification and Fingerprints Construction of Junlongyou No.5 in Brassica napus

利用SSR和SRAP分子标记技术分别对甘蓝型油菜细胞质雄性不育均隆油5号亲本和F1代进行研究,从240对SSR引物、255对SRAP引物中分别筛选出11对和8对多态性好且子代含父母本特征条带的引物,从中选出5对SSR引物(CB10057、Na10-D03 A、Na 12-E 02、CB 10364、CB 10179)和5对SRAP引物(Me 1-Em 10、Me 6-Em 3、Me 6-Em 11、Me 6-Em 19、Me 6-Em 20)作为核心引物,用于F1代杂种纯度鉴定。结果表明,SSR和SRAP两种分子标记所测杂交种纯度分别为97.7%和100%,与田间纯度调查结果具有高度的一致性。从11个SSR标记和8个SRAP标记中分别选出5个SSR和3个SRAP标记组成分子标记组合,构建了均隆油5号油菜亲本、F1代及其它市售42份油菜品种的特征指纹图谱,实现了对不同品种的特异性鉴定。在此基础上建立的数字指纹图谱比对程序实现了对特定品种(品系)指纹图谱的比对分析,可作为杂种纯度分析及假冒伪劣种子判定的参考。

In this study,SSR and SRAP markers were used to analyze the cytoplasmic male sterility（CMS） of the parents and F1 of Junlongyou No.5 in Brassica napus.11 out of 240 pairs of SSR primers and 8 out of 255 pairs of SRAP primers were selected for their high polymorphism and the offspring contain the special bands of their parents.Five SSR primers including CB 10057 、Na 10-D 03 A、Na 12-E 02 、CB 10364 、CB 10179 and five SRAP primers including Me 1-Em 10、Me 6-Em 3、Me 6-Em 11 、Me 6-Em 19、Me 6-Em 20 were selected as hybrid purity identification of F1.The results showed that the seed purity test by SSR and SRAP markers were 97.7％ and 100％,respectively,which had high consistent with the test in field.5 out of 11 pairs of SSR primers and 3 out of 8 pairs of SRAP primers were selected to build the special DNA fingerprinting database of the parents and F1 of Junlongyou No.5 in Brassica napus and other 42 rapeseed varieties.The fingerprint matching program based on the database can realize the comparison analysis of fingerprints of the specific varieties （strains）,which can also used to the reference of hybrid purity identification and the judgment of fake seeds.