射干内生真菌Fusarium proliferatum的ISSR和SRAP位点遗传多样性分析  

Genetic diversity of endophytic fungi strains Fusarium proliferatum isolated from Belamcanda chinensis using ISSR and SRAP markers

作　　者：罗琴 赵欢 彭正松 李佩华 杨玉霞 夏燕莉 邓迪 LUO Qin;ZHAO Huan;PENG Zheng-song;LI Pei-hua;YANG Yu-xia;XIA Yan-li;DENG Di(College of Life Science,China West Normal University,Nanchong 637009,China;Xichang College,Xichang 615013,China;Sichuan Academy of Traditional Chinese Medicine Sciences,Chengdu 625014,China)

机构地区：[1]西华师范大学生命科学学院 [2]西昌学院 [3]四川省中医药科学院

出　　处：《中草药》2019年第23期5847-5857,共11页Chinese Traditional and Herbal Drugs

基　　金：四川省教育厅重大培育项目（15CZ0015）;西华师范大学博士启动基金（15E025）;西华师范大学英才科研基金项目（17YC364）;四川省大学生创新创业项目（201810638100）

摘　　要：目的为了解射干内生真菌Fusarium proliferatum的遗传多样性。方法选用了52条ISSR引物和90条SRAP引物对17株射干内生真菌F.proliferatum进行了遗传多样性分析。筛选出了27条ISSR引物和38条SRAP引物可用于遗传多样性分析。结果结果,27条ISSR引物共扩增出了178个条带,其中131条(63%)具有多态性;38条SRAP引物能够扩增出357条稳定性较好的条带,其中323条(91%)具有稳定性差异。27条ISSR引物PCR扩增产物的多态性信息量(PIC)介于0.19~0.91,平均为0.70;38条SRAP引物PCR扩增产物的多态性信息量PIC介于0~0.93,平均为0.73。聚类分析结果表明,根据27个ISSR、38个SRAP及ISSR+SRAP遗传位点分析得出遗传相似系数变化范围分别为0.73~0.99、0.72~0.95和0.73~0.95,平均分别为0.84、0.85和0.85。根据材料间的遗传相似系数聚类,三者之间的聚类有较小的差异,但总体上,17株内生真菌被聚为3大类,分离自根的内生真菌F.proliferatum聚为一类,分离自茎和叶的F.proliferatum聚集于另外两类。研究表明,内生真菌F.proliferatum遗传相似性较大,其亲缘关系较近,与传统方法鉴定出的结果一致。结论采用分子标记技术比传统的方法鉴定更为有效,同时,ISSR和SRAP标记可更真实地反映射干内生真菌F.proliferatum的遗传多样性,为该内生真菌在分子生物技术方面的研究等提供一定参考。Objective In order to understand the genetic diversity of endophytic fungi strain Fusarium proliferatum isolated from Belamcanda chinensis.Methods A total of 52 ISSR primers and 90 SRAP primers were used to detect the genetic diversity among 17 F.proliferatum strains.Results The results indicated that 27 ISSR primers and 38 SRAP primers were screened out for the genetic diversity analysis.178 bands were amplified from 27 ISSR primers,among which 131(63%)allelic variations were detected.However,357 bands were amplified by 38 SRAP primers,among which 323(91%)allelic variations were detected.The value of allelic polymorphism information content(PIC)of ISSR primers ranged from 0.19 to 0.91,with the average of 0.70 per primer.The value of PIC of SRAP primers ranged from 0.00 to 0.93,with the average of 0.72 per primer.The value of Nei's genetic similarity(GS)indexes of 17 strains based on ISSR,SRAP and ISSR+SRAP genetic locus varied from 0.73-0.99,0.72-0.95 and 0.73-0.95,and with the average of 0.84,0.85 and 0.85,separately.Cluster analysis showed that the 17 strains in this study could be clustered into three groups,three strains from the roots were clustered together,and F.proliferatum strains isolated from stems and leaves were gathered in other two groups.Cluster analysis revealed that genetic similarity of 17 strains were high,this suggested that the 17 strains had a near relationship,in accordance with the traditional morphology identification.Conclusion The results show that the ISSR and SRAP technology is more efficient than traditional morphology identification.It is also found that ISSR and SRAP markers could more really reflect the genetic diversity of endophytic fungi strain F.proliferatum from B.chinensis,which can provide the basis for the application of molecular biotechnology in endophytic fungi of F.proliferatum from B.chinensis.

关 键 词：射干 内生真菌 Fusarium proliferatum 遗传多样性 ISSR SRAP 

分 类 号：R282.12[医药卫生—中药学]