Ouyang Huili, Wang Xiaoyun, Zheng Xialin, Lu Wen, Qin Fengping, Chen Chao
Guangxi Key Laboratory of Agric-Environment and Agric-Products Safety, College of Agriculture, Guangxi University, Nanning 530004, China.
Insects. 2021 Oct 14;12(10):938. doi: 10.3390/insects12100938.
(Hendel), as one of the most notorious and destructive invasive agricultural pests in the world, causes damage to over 250 different types of fruits and vegetables throughout tropical and subtropical areas. PacBio single-molecule real-time (SMRT) sequencing was used to generate the full-length transcriptome data of . A total of 40,319,890 subreads (76.6 Gb, clean reads) were generated, including 535,241 circular consensus sequences (CCSs) and 386,916 full-length non-concatemer reads (FLNCs). Transcript cluster analysis of the FLNC reads revealed 22,780 high-quality reads (HQs). In total, 12,274 transcripts were functionally annotated based on four different databases. A total of 1978 SSR loci were distributed throughout 1714 HQ transcripts, of which 1926 were complete SSRs and 52 were complex SSRs. Among the total SSR loci, 2-3 nucleotide repeats were dominant, occupying 83.62%, of which di- and tri- nucleotide repeats were 39.38% and 44.24%, respectively. We detected 105 repeat motifs, of which AT/AT (50.19%), AC/GT (39.15%), CAA/TTG (32.46%), and ACA/TGT (10.86%) were the most common in di- and tri-nucleotide repeats. The repeat SSR motifs were 12-190 bp in length, and 1638 (88.02%) were shorter than 20 bp. According to the randomly selected microsatellite sequence, 80 pairs of primers were designed, and 174 individuals were randomly amplified by PCR using primers. The number of primers that had amplification products with clear bands and showed good polymorphism came to 41, indicating that this was a feasible way to explore SSR markers from the transcriptomic data of . These results lay a foundation for developing highly polymorphic microsatellites for researching the functional genomics, population genetic structure, and genetic diversity of .
(亨德尔)作为世界上最臭名昭著、破坏力最强的入侵性农业害虫之一,在热带和亚热带地区对250多种不同类型的水果和蔬菜造成损害。使用PacBio单分子实时(SMRT)测序来生成……的全长转录组数据。共产生了40319890条子读数(76.6 Gb,clean reads),包括535241条环状一致序列(CCSs)和386916条全长非串联读数(FLNCs)。对FLNC读数进行转录本聚类分析,发现了22780条高质量读数(HQs)。总共基于四个不同数据库对12274条转录本进行了功能注释。共有1978个SSR位点分布在1714条HQ转录本中,其中1926个是完整的SSR,52个是复杂的SSR。在总的SSR位点中,2 - 3核苷酸重复占主导,占83.62%,其中二核苷酸和三核苷酸重复分别占39.38%和44.24%。我们检测到105个重复基序,其中AT/AT(50.19%)、AC/GT(39.15%)、CAA/TTG(32.46%)和ACA/TGT(10.86%)在二核苷酸和三核苷酸重复中最为常见。重复SSR基序的长度为12 - 190 bp,1638个(88.02%)短于20 bp。根据随机选择的微卫星序列设计了80对引物,并使用引物通过PCR对174个个体进行随机扩增。具有清晰条带且显示出良好多态性的引物数量为41个,表明这是从……的转录组数据中探索SSR标记的可行方法。这些结果为开发高度多态的微卫星用于研究……的功能基因组学、群体遗传结构和遗传多样性奠定了基础。