适于柚×橙杂交后代遗传分析的ISSR引物的筛选与应用

Screening and Application of ISSR Primers for Hybrid Progenies from Citrus grandis×C. sinensis

以沙田柚、强德勒柚、红江橙为材料,利用优化的柑橘ISSR体系,对100个ISSR引物进行了筛选.结果表明：28个引物能够扩增出2~12条稳定、清晰的条带,多态性条带为1~9条,多态率为25%~90%,其中P807,P808,P809,P810,P811,P812,P814,P815,P820,P821,P822,P826,P827,P830,P834,P835,P840,P845等18个引物在沙田柚和红江橙之间有差异,P812,P814,P815,P827,P834,P840等6个引物在沙田柚和强德勒柚之间有差异.用2个差异引物P827和P834可从36株沙田柚×强德勒柚杂交后代中鉴定出31株为真杂种,用P810,P811,P812,P835等4个引物可以把52株沙田柚×红江橙杂种后代全部鉴定出来,并且获得了一个适用于沙田柚和红江橙杂交后代鉴定分析的完全显性标记P810.

One hundred ISSR primers were used for ISSR-PCR amplification in three citrus varieties,Shatian pummelo（Citrus grandis）,Chandler pummelo（C.grandis） and Hongjiangcheng（C.sinensis）.Twenty-eight primers were effective,each of which amplified 2-12 stable and clear bands,of which 1-9 bands were polymorphic,with a polymorphic allele rate of 25%-90%.Eighteen primers（P807,P808,P809,P810,P811,P812,P814,P815,P820,P821,P822,P826,P827,P830,P834,P835,P840 and P845） could discriminate Shatian pummelo and Chandler pummelo,and 6 primers（P812,P814,P815,P827,P834 and P840） showed differences between Shatian pummelo and Hongjiangcheng.Thirty-one hybrids were identified from 36 F1 seedlings of Shatianyou × Chandler pummelo by primer P827 and P834,all the offspring from Shatianyou × Hongjiangcheng were proved to be hybrids with primers P810,P811,P812 and P835,and one homozygous dominant marker P810 was obtained.