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Gluten hydrolase gene silencing using RNAi and its effect on the Sunn pest growth and development
Phytoparasitica ( IF 1.4 ) Pub Date : 2020-06-05 , DOI: 10.1007/s12600-020-00821-8
Azam Amiri , Ali R. Bandani

RNAi technology has been described as a tool to study gene function and is of great potential as a next-generation biopesticide. Thus, in the present study, we identified and sequenced the Sunn pest Gluten hydrolase (GH) gene partially. Then, evaluated GH-dsRNA effects on the GH expression of the fifth instar nymphs and adults, as well as, on the insect growth and development using topical and injection assays. Results indicated that the topical application of GH-dsRNA on the fifth instar nymph caused a severe reduction (99.9%) of gut GH expression in comparison with control and, at the same time, high induction of the salivary gland’s expression profile. Gene upregulation indicates that the insect compensates for the reduction of the one gene expression with the induction of a gene in the other tissue to meet its demand. Also, the GH expression profile in the gut and salivary glands were evaluated 6, 12, and 21-hour post injections. Results revealed that the GH expression profile of both gut and salivary glands was reduced early hours after injection, but their expression profile was recovered 21-hour post-treatment. Thus, the results showed that dsRNA does not have a systemic effect since gene knockdown was not persistent. GH-dsRNA also affected nymphal development time, adult weight, survivability, and adult emergence. These effects indicated that the GH gene plays an essential physiological role(s) in addition to having a role in food digestion. Therefore, RNAi technology has the potential to be used to develop novel strategies for Sunn pest IPM programs.

中文翻译:

利用 RNAi 沉默谷蛋白水解酶基因及其对 Sunn 害虫生长发育的影响

RNAi 技术已被描述为研究基因功能的工具,并且作为下一代生物农药具有巨大潜力。因此,在本研究中,我们对 Sunn 害虫面筋水解酶 (GH) 基因进行了部分鉴定和测序。然后,使用局部和注射试验评估了 GH-dsRNA 对五龄若虫和成虫 GH 表达的影响,以及对昆虫生长和发育的影响。结果表明,与对照相比,在五龄若虫上局部应用 GH-dsRNA 导致肠道 GH 表达严重降低 (99.9%),同时,唾液腺表达谱的高度诱导。基因上调表明昆虫通过在另一个组织中诱导基因来补偿一个基因表达的减少以满足其需求。还,在注射后 6、12 和 21 小时评估肠道和唾液腺中的 GH 表达谱。结果显示,肠道和唾液腺的 GH 表达谱在注射后数小时内降低,但它们的表达谱在治疗后 21 小时恢复。因此,结果表明 dsRNA 不具有全身效应,因为基因敲低不是持久的。GH-dsRNA 还影响若虫发育时间、成虫体重、存活率和成虫羽化。这些影响表明 GH 基因除了在食物消化中发挥作用外,还发挥着重要的生理作用。因此,RNAi 技术有可能用于开发 Sunn 害虫 IPM 计划的新策略。结果显示,肠道和唾液腺的 GH 表达谱在注射后数小时内降低,但它们的表达谱在治疗后 21 小时恢复。因此,结果表明 dsRNA 不具有全身效应,因为基因敲低不是持久的。GH-dsRNA 还影响若虫发育时间、成虫体重、存活率和成虫羽化。这些影响表明 GH 基因除了在食物消化中发挥作用外,还发挥着重要的生理作用。因此,RNAi 技术有可能用于开发 Sunn 害虫 IPM 计划的新策略。结果显示,肠道和唾液腺的 GH 表达谱在注射后数小时内降低,但它们的表达谱在治疗后 21 小时恢复。因此,结果表明 dsRNA 不具有全身效应,因为基因敲低不是持久的。GH-dsRNA 还影响若虫发育时间、成虫体重、存活率和成虫羽化。这些影响表明 GH 基因除了在食物消化中发挥作用外,还发挥着重要的生理作用。因此,RNAi 技术有可能用于开发 Sunn 害虫 IPM 计划的新策略。结果表明,dsRNA 不具有全身效应,因为基因敲低不是持续性的。GH-dsRNA 还影响若虫发育时间、成虫体重、存活率和成虫羽化。这些影响表明 GH 基因除了在食物消化中发挥作用外,还发挥着重要的生理作用。因此,RNAi 技术有可能用于开发 Sunn 害虫 IPM 计划的新策略。结果表明,dsRNA 不具有全身效应,因为基因敲低不是持续性的。GH-dsRNA 还影响若虫发育时间、成虫体重、存活率和成虫羽化。这些影响表明 GH 基因除了在食物消化中发挥作用外,还发挥着重要的生理作用。因此,RNAi 技术有可能用于开发 Sunn 害虫 IPM 计划的新策略。
更新日期:2020-06-05
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