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对罗氏沼虾(Macrobrachium rosenbergii)内转录间隔区(internal transcribed spacer,ITS)全序列特征进行了分析。罗氏沼虾ITS1长度为1 070～1 150 bp,GC含量为51.4%～52.7%;5.8S长度一致为163bp,GC含量为55.2%;ITS2长度为484～545 bp,GC含量为53.4%～54.6%。序列比对显示,ITS1存在61处SNPs,其中C/T转换为28处(占45.9%),A/G转换为23处(占37.7%),A/T颠换为1处(占1.6%),T/G颠换为3处(占4.9%),C/G颠换为4处(占6.6%),A/C颠换为2处(占3.3%);ITS2存在15处SNPs,其中C/T转换为6处(占40%),A/G转换为5处(占33.3%),A/T颠换为1处(占6.7%),T/G颠换为2处(占13.3%),C/G颠换为1处(占6.7%)。整个ITS中微卫星序列(SSR)共有20处,其中ITS1中有14处,以(AG)n类型最多;ITS2中有6处,以(GA)n类型最多。人工查找结果发现,罗氏沼虾ITS1在181、390和781位点存在3处59 bp正向重复单元。个体内差异分析表明,罗氏沼虾ITS由于SSR重复数不同,而存在个体内差异。同源性分析表明,罗氏沼虾ITS1与海南沼虾(M.hainanense)同源性最高,为50.64%,而与脊尾白虾(Exopalaemon carinicauda)同源性最低,为45.39%;5.8S与其他甲壳动物有着极高的同源性,其中与日本沼虾(M.nipponense)同源性达到100%;ITS2与日本沼虾同源性最高,为53.46%,而与克氏原螯虾(Procambarus clark)的同源性最低,为42.53%。基于ITS构建的NJ树显示,罗氏沼虾与同属的日本沼虾聚为一支,与传统分类相符。
The complete sequences of internal transcribed spacer (ITS) sequences in Macrobrachium rosenbergii were analyzed. The ITS1 length ranged from 1 070 to 1 150 bp and the GC content from 51.4% to 52.7%. The length of 5.8S was 163bp, the content of GC was 55.2%. The length of ITS2 was from 484 to 545 bp and the GC content was from 53.4% to 54.6 %. There were 61 SNPs in ITS1, of which 28 were C / T (45.9%), 23 were A / G (37.7%) and 1 was A / T (1.6%), ), T / G translocation to 3 (4.9%), C / G translocation to 4 (6.6%) and A / C translocation to 2 (3.3% Among them, C / T was converted to 6 (40%), A / G converted to 5 (33.3%), A / T transposed to 1 Accounting for 13.3%), C / G translocation to 1 (6.7%). There were 20 microsatellite loci (SSRs) in ITS, of which 14 were in ITS1, the most was (AG) n, and the other 6 were ITS2 (GA) n. The results of the manual search revealed that there are three 59 bp forward repeat units in ITS1, 181, 390 and 781 loci of M. rosenbergii. In-house variability analysis showed that ITS in M. rosenbergii varied in-house because of SSR numbers. Homology analysis showed that M. rosenbergii ITS1 had the highest homology with M. hainanense (50.64%), but the lowest homology with Exopalaemon carinicauda (45.39%). 5.8S was associated with other The crustacean has extremely high homology, of which the homology with M. nipponense reaches 100%. ITS2 has the highest homology with M. marsh, accounting for 53.46%, while the homology with Procambarus clark ) Had the lowest homology of 42.53%. The NJ tree constructed based on ITS showed that Macrobrachium rosenbergii and Macrobrachium nipponensis were clustered together and accorded with the traditional classification.