Changes in transcript abundance of 24 genes expressed in the dorsal pharyngeal gland cell of Heterodera glycines encoding for putative secretions of unknown function were monitored by quantitative PCR (qPCR) at 0, 2, 7, 14 and 21 days post-invasion (pi) of soybean plantlets. Five groups of temporal patterns (A, B1, B2, C and D) were defined for the 24 genes plus data for two previously studied genes expressed in the same cell. Group D (two genes) showed no significant increase between 0 and 2 days pi and were the least abundantly expressed at 7-21 days pi. Transcripts of group C (nine genes including one studied previously) increased in abundance from 0 to 2 days pi but were the second least expressed for 7-21 days pi. Groups A (three genes), B1 (seven genes) and B2 (five genes including one studied previously) were all abundant at 7-21 days pi. B1 and B2 were discriminated by their relative abundance at 0 and 2 days pi. RNA interference (RNAi) targeting two genes of group A and one each of B1 and B2 in nematodes prior to invasion resulted in phenotypic effects on total parasites per plant and sexual fate at 10 days pi. Phenotype penetrance was reduced for three genes showing such effects and one with a strong effect in earlier work when two genes rather than one were concurrently targeted for RNAi. One gene (dg13) was more abundantly expressed after combinatorial RNAi than for either control nematodes or when targeting singly by RNAi. This work reports the unexpected elevation in mRNA expression after combinatorial RNAi that requires understanding before combinatorial RNAi can be advanced for highly effective cyst nematode control via plant biotechnology.

中文翻译：

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大豆胞囊线虫背咽腺细胞中表达的先锋基因的 RNAi 表征以及组合 RNAi 的影响。


在大豆胞囊线虫入侵后 0、2、7、14 和 21 天 (pi)，通过定量 PCR (qPCR) 监测编码未知功能的推定分泌物的 24 个基因转录本丰度的变化，这些基因在大豆胞囊线虫背咽腺细胞中表达。大豆苗。为 24 个基因以及先前研究的在同一细胞中表达的两个基因的数据定义了五组时间模式（A、B1、B2、C 和 D）。 D 组（两个基因）在感染后 0 至 2 天之间没有表现出显着增加，并且在感染后 7-21 天时表达量最低。 C组的转录本（九个基因，包括之前研究过的一个）在感染后0至2天内丰度增加，但在感染后7-21天表达量第二低。 A组（三个基因）、B1组（七个基因）和B2组（五个基因，包括之前研究过的一个基因）在感染后7-21天都丰富。 B1和B2通过它们在感染后第0天和第2天的相对丰度来区分。在入侵之前，针对线虫中 A 组的两个基因以及 B1 和 B2 各一个基因的 RNA 干扰 (RNAi) 会对每株植物的寄生虫总数和感染后 10 天的性命运产生表型影响。在早期研究中，当同时针对 RNAi 的两个基因而不是一个基因时，表现出这种效应的三个基因和一个具有强烈效应的基因的表型外显率降低。组合 RNAi 后，一个基因 (dg13) 的表达量比对照线虫或 RNAi 单独靶向时的表达更为丰富。这项工作报告了组合 RNAi 后 mRNA 表达的意外升高，需要先了解组合 RNAi 才能通过植物生物技术进行高效的胞囊线虫控制。