昆虫学报2017，Vol.60Issue(5)：582-593,12.DOI:10.16380/j.kcxb.2017.05.011
中国新发现芙新姬小蜂孤雌产雌品系的分子鉴定及其内共生菌Rickettsia的检测
Molecular identification of the thelytokous strain of Neochrysocharis formosa (Hymenoptera: Eulophidae) newly found in China and detection of its endosymbiont Rickettsia
摘要
Abstract
[Aim] The eulophid wasp Neochrysocharis formosa (Westwood) is an important dominant parasitoid against vegetable agromyzid leafminers all over the world.In June 2015,we found the thelytokous strain of N.formosa in Beijing,which was its first discovery in China.It is difficult to distinguish thelytokous strain from arrhenotokous strain because of no distinct morphological differentiation.In this study molecular methods were adopted to identify these two strains so as to provide a basis for future research and application of the thelytokous strain of N.formosa.[Methods] The genetic differentiation between the thelytokous strain TH-China and the arrhenotokous strain AR-China of N.formosa reared for several generations indoor was analyzed by multiple molecular markers (COⅠ gene,ITS-1 gene and 28S rDNA).The 16S rDNA sequence was amplified from the endosymbiont Rickettsia of the two strains using the specific primers NforRickl/NforRick2,sequenced and blasted against NCBI database.The existence of Rickettsia which can induce thelytoky was detected in ovarioles,larvae,pupae and adults of TH-China,and adults of AR-China.[Results] Phylogenetic trees constructed based on two fragments of COⅠ gene amplified with the primer pairs LCO1490/HCO2198 and COⅠ1/COⅠ2 showed that the thelytokous and arrhenotokous strains of N.formosa diverged into two distinct clades and their genetic distance is 0.039 and 0.023,respectively.Using the ITS-1 gene and 28S rDNA as the molecular markers,there are 21 and 0 discrepant sites between the two strains,respectively,and the genetic distance of the two strains based on ITS-1 gene is 0.008.The 16S rDNA sequence amplified from Rickettsia in TH-China strain has 100％ identity with that from Rickettsia in the thelytokous strain in Japan (GenBank accession no.:AB185963).Rickettsia was detected in ovarioles,larvae,pupae and adults of the thelytokous strain,but absent in adults of the arrhenotokous strain,suggesting that Rickettsia might contribute to the thelytoky.[Conclusion] The thelytokous and arrhenotokous strains of N.formosa are easily differentiated by COⅠ gene sequences,while no genetic variation is found based on 28S rDNA and almost no genetic divergence based on ITS-1 gene.关键词
芙新姬小蜂/孤雌产雌品系/两性品系/分子鉴定/Rickettsia/COⅠ/ITS-1/28S rDNAKey words
Neochrysocharis formosa/thelytokous strain/arrhenotokous strain/molecular identification/Rickettsia/COⅠ/ITS-1/28S rDNA分类
生物科学引用本文复制引用
杨月梅,轩景丽,叶福宇,郭建洋,杨利艳,刘万学..中国新发现芙新姬小蜂孤雌产雌品系的分子鉴定及其内共生菌Rickettsia的检测[J].昆虫学报,2017,60(5):582-593,12.基金项目
北京市自然科学基金项目(6142016) （6142016）
国家重点研发计划项目(2016YFC1201200) （2016YFC1201200）
国家自然科学基金项目(31372005) （31372005）
