RAPD技术在木耳属杂交种及原生质体融合子鉴定分析中的应用

THE USE OF RAPD IN IDENTIFICATION OF INTRASPECIES HYBRIDS AND INTERSPECLES PROTOPLAST FUSANTS IN AU RICULLARIA

本研究以ＲＡＰＤ（ＲａｎｄｏｍｌｙＡｍｐｌｉｆｉｅｄＰｏｌｙ－ｍｏｒｐｈｉｃＤＮＡ）技术为主，比较分析食用菌木耳属（Ａｕｒｉｃｕｌａｒｉａ）种内杂交种．种间原生质体融合子与其亲本菌株的ＰＣＲ（ＰｏｌｙｍｅｒｓｅＣｈａｉｈＲｅａｃｔｉｏｎ）扩增的ＤＮＡ片段之间差异，并辅以其它手段，如核相及核数目观察，同功酶鉴别，以期建立完整有效的检测技术系列，确证其杂交种融合子的真实性及可靠性。在ＲＡＰＤ分析中，用于测试的１０种随机引物仅有４种引物与供试菌株的ＤＮＡ模板具有较好的结合能力，其中１种引物可用于检测杂交种及其亲本，另外３种引物可用于检测融合子反亲本。检测结果显示：３个供试杂交种中有２个为真杂合子，１个为假杂合子；２个供试融合子均为非稳定杂合子。与常规方法鉴定结果比较基本相符。因此，ＲＡＰＤ技术提供了一种快速、简便、准确的杂交种及融合子的鉴别方法。

RAPD(Randomy Amplified Polymorphic DNA)was used to analysethe differences of DNA fragments between intraspecies hybrid,interspecies protoplastfusants and their respective parents in genus Auricularia,With the help of other methodssuch as the nuclear number in the hyphal cell, the comparison of the isozyme graphs, the hybrids and the protoplast fusants were identified.Four of ten arbitrary primets had good compatibility with DNA template of test strainsin RAPD assay. One of four was used to dentify by brids and their parents, three of fourwere used to identify fusants and their parents. The results showed that two of threehybrids were real heterozygotes,the two fusants were not stable.Therefore,comparing with the other ways,RAPD provided a rapid, simple, accurateidentified method of hybrids and fusants.