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首页> 外文期刊>Planta: An International Journal of Plant Biology >Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe
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Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe

机译：宿主对线虫生殖和适应基因的抑制会降低异型异型甘氨酸Ichinohe的繁殖力

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摘要

To control Heterodera glycines Ichinohe (soybean cyst nematode) in Glycine max (L.) Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reproduction or fitness (Cpn-1, Y25 and Prp-17) were PCR-amplified using specific primers and independently cloned into the pANDA35HK RNAi vector using a Gateway cloning strategy. Soybean roots were transformed with these constructions using a composite plant system. Confirmation of transformation was attained by PCR and Southern blot analysis. Transgene expression was detected using reverse transcription PCR (RT-PCR) and expression of siRNAs was confirmed in transgenic plants using northern blot analysis. Bioassays performed on transgenic composite plants expressing double-stranded RNA fragments of Cpn-1, Y25 and Prp-17 genes resulted in a 95, 81 and 79% reduction for eggs g~(-1) root, respectively. Furthermore, we demonstrated a significant reduction in transcript levels of the Y25 and Prp-17 genes of the nematodes feeding on the transgenic roots via real-time RT-PCR whereas the expression of non-target genes were not affected. The results of this study demonstrate that over-expression of RNA interference constructs of nematode reproduction or fitness-related genes can effectively control H. glycines infection with levels of suppression comparable to conventional resistance.
机译：为了控制最大大豆（L.）Merr。中的异型藻甘氨酸Ichinohe（大豆孢囊线虫）。 （大豆），我们评估了生产表达针对特定H.甘氨酸基因的小干扰RNA（siRNA）的转基因大豆幼苗的用途。使用特定引物对与线虫繁殖或适应性相关的三个基因（Cpn-1，Y25和Prp-17）的基因片段进行PCR扩增，并使用Gateway克隆策略将其独立克隆到pANDA35HK RNAi载体中。使用复合植物系统，用这些结构转化大豆根。通过PCR和Southern印迹分析获得转化的确认。使用逆转录PCR（RT-PCR）检测转基因表达，并使用northern blot分析在转基因植物中确认siRNA的表达。对表达Cpn-1，Y25和Prp-17基因双链RNA片段的转基因复合植物进行的生物测定分别使鸡蛋g〜（-1）根减少了95％，81％和79％。此外，我们证明通过实时RT-PCR取食转基因根的线虫的Y25和Prp-17基因的转录水平显着降低，而非靶标基因的表达不受影响。这项研究的结果表明，线虫繁殖或适应性相关基因的RNA干扰构建体的过表达可以有效地控制H.甘氨酸感染，其抑制水平可与常规抗药性媲美。

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