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期 刊 ：植物保护学报 2009年

关键词：昆虫病原线虫；索科；tra-1基因；3-RACE；实时荧光定量PCR；

Keywords:entomopathogenic nematodes, Mermithidae, tra-1 gene, 3‘-RACE, real-time quantitative RT-PCR,

摘 要 ：利用RT-PCR和3‘-RACE,从中华卵索线虫Ovomermis sinensis中获得tra-1基因cDNA部分序列,该序列长800bp,共编码195个氨基酸,与GLI转录因子家族其它成员在锌指区域具有较高的相似性(70%~77%)。实时荧光定量检测显示:雌成虫tra-1相对表达量最高,与其它虫期差异极显著(P<0.01),在怀卵雌虫和胚胎发育期卵中,tra-1相对表达量明显下降,但仍显著高于其它试验组(P<0.05);其它试验组,即感染期幼虫、1~6天寄生期幼虫、寄生后期幼虫与雄成虫之间表达量无明显差异(P>0.05)。线虫在宿主体内不同的感染强度(40/1与10/1)对tra-1基因转录活性无显著影响(P>0.05)。表明tra-1基因参与调控中华卵索线虫雌性生殖系统的发育、卵的形成以及胚胎的发育。

Abstract:Tra-1 gene was partially cloned from mermithid nematode by using RT-PCR and 3‘-RACE for the first time. The 5‘ truncated cDNA was 800bp in length and encoded a putative protein of 195 amino acids, which had high homology with other members of GLI zinc-finger transcription factors in the zinc-finger domain. The identities were 70%-77%. The transcript levels of tra-1 were analyzed by real-time fluorescent quantitative PCR. The results showed that the tra-1 transcript presented high levels in the female adults (P<0.01), then dropped observably during the stages of embryos and mated female adults, while were significantly higher than that in other groups (P<0.05). However, the expressions among pre-parasites, parasites, post-parasites and male adults, had no remarkable differences (P>0.05). Different numbers of nematode hosted in host didn’t infect the tra-1’s transcription obviously (P>0.05). Studying on the basis of quantitative results, the author presumed the TRA-1A had a share in the control of the gonad development, the oogenesis and embryonic development of the O.sinensis.

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