Deng Zhongyuan, Zhang Yakun, Zhang Min, Huang Jinyong, Li Changyou, Ni Xinzhi, Li Xianchun
School of Agricultural Sciences, Zhengzhou University, Zhengzhou, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
Front Genet. 2020 Jun 19;11:649. doi: 10.3389/fgene.2020.00649. eCollection 2020.
is a globally-important crop pest with a WZ (female)/ZZ (male) sex chromosome system. The absence of discernible sexual dimorphism in its egg and larval stages makes it impossible to address any sex-related theoretical and applied questions before pupation unless a W-specific sequence marker is available for sex diagnosis. To this end, we used one pair of morphologically pre-sexed pupae to PCR-screen 17 non-transposon transcripts selected from 4855 W-linked candidate reads identified by mapping a publicly available egg transcriptome of both sexes to the male genome of this species and detected the read SRR1015458.67499 only in the female pupa. Subsequent PCR screenings of this read and the previously reported female-specific RAPD (random amplified polymorphic DNA) marker with ten more pairs of pre-sexed pupae and different annealing positions and/or temperatures as well as its co-occurrence with the female-specific transcript splicing isoforms of gene of and amplification and sequencing of their 5' unknown flanking sequences in three additional pairs of pre-sexed pupae verified that SRR1015458.67499 is a single copy protein-coding gene unique to W chromosome (named ) while is a multicopy MITE transposon located on various chromosomes. Test application of as a marker to sex 30 neonates of yielded a female/male ratio of 1.14: 1.00. Both and splicing isoforms assays revealed that the embryo cell line QB-Ha-E-1 is a male cell line. Taken together, is not only a reliable DNA marker for sexing all stages of and its cell lines, but also represents the first W-specific protein-coding gene in lepidopterans.
是一种具有WZ（雌性）/ZZ（雄性）性染色体系统的全球重要作物害虫。其卵和幼虫阶段缺乏明显的性二态性，这使得在化蛹前无法解决任何与性别相关的理论和应用问题，除非有W特异性序列标记可用于性别诊断。为此，我们使用一对形态学上预先确定性别的蛹，对从4855个W连锁候选reads中选出的17个非转座子转录本进行PCR筛选，这些候选reads是通过将该物种雌雄两性的公开可用卵转录组映射到雄性基因组而确定的，并且仅在雌性蛹中检测到reads SRR1015458.67499。随后，用另外十对预先确定性别的蛹、不同的退火位置和/或温度对该reads以及先前报道的雌性特异性RAPD（随机扩增多态性DNA）标记进行PCR筛选，以及在另外三对预先确定性别的蛹中其与基因的雌性特异性转录本剪接异构体的共现及其5'未知侧翼序列的扩增和测序，证实SRR1015458.67499是W染色体特有的单拷贝蛋白质编码基因（命名为），而 是位于各种染色体上的多拷贝MITE转座子。将作为标记物用于对30只 的新生幼虫进行性别鉴定的测试应用产生的雌雄比为1.14:1.00。 和 剪接异构体分析均表明胚胎细胞系QB-Ha-E-1是雄性细胞系。综上所述，不仅是用于对 及其细胞系的所有阶段进行性别鉴定的可靠DNA标记，而且代表鳞翅目中第一个W特异性蛋白质编码基因。
