柑桔RAPD技术体系建立与体细胞杂种鉴定

Establishment of RAPD Analysis Techniques and Identification of Somatic Hybrids in Citrus

通过优化反应条件，建立起柑桔ＲＡＰＤ分析技术体系，并成功地应用于体细胞杂种鉴定。柑桔ＲＡＰＤ以Ｍｇ２＋为１．５ｍｍｏｌ／Ｌ、〔ｄＮＴＰｓ〕为１００μｍｏｌ／Ｌ时扩增稳定，效果良好。用１６个引物扩增〔哈姆林甜橙（＋）粗柠檬〕融合植株，体细胞杂种表现为双亲谱带之和。结合染色体计数，可将〔Ｐａｇｅ桔柚（＋）粗柠檬〕融合再生的体细胞杂种（异源四倍体）、叶肉亲本类型及同源融合植株区别开来。

RAPD analysis techniques for Citrus were established, and successfully used for somatic hybrid identification. The good results could be obtained when 1.5mmol/ L Mg 2+ and 100μmol/ L dNTPs were used in the system. Amplified with 16 arbitrary primers, the somatic hybrids from ‘Hamlin sweet orange’ and ‘Rough lemon’ presented the total bands of 2 parents. Combined with chromosome counting, somatic hybrids (allotetraploids) , leaf-parent-type regenerants and autotetraploids from ‘Page’ tangelo and ‘Rough lemon’ could be verified.