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向疫霉属晚疫病菌喷洒 dsRNA 分子,并与纳米黏土载体一起作为开发马铃薯晚疫病新型保护策略的概念验证。

Spraying of dsRNA molecules derived from Phytophthora infestans, along with nanoclay carriers as a proof of concept for developing novel protection strategy for potato late blight.

机构信息

Plant Protection, ICAR-Central Potato Research Institute, Shimla, India.

Plant Biochemistry, ICAR-Indian Grassland and Fodder Research Institute, Jhansi, India.

出版信息

Pest Manag Sci. 2022 Jul;78(7):3183-3192. doi: 10.1002/ps.6949. Epub 2022 May 18.

Abstract

BACKGROUND

Phytophthora infestans is a late blight-causing oomycetes pathogen. It rapidly evolves and adapts to the host background and new fungicide molecules within a few years of their release, most likely because of the predominance of transposable elements in its genome. Frequent applications of fungicides cause environmental concerns. Here, we developed target-specific RNA interference (RNAi)-based molecules, along with nanoclay carriers, that when sprayed on plants are capable of effectively reducing late blight infection.

RESULTS

Targeted the genes unique to sporulation, early satge infection and the metabolism pathway stages based on in an our own microarray data. We used nanoclay as a carrier for sorbitol dehydrogenase, heat shock protein 90, translation elongation factor 1-α, phospholipase-D like 3 and glycosylphosphatidylinositol-anchored acidic serine-threonine-rich HAM34-like protein double-stranded (ds)RNAs, which were assessed by culture bioassay, detached leaf assay and spray methods, and revealed a reduction in growth, sporulation and symptom expression. Plants sprayed with multigene targeted dsRNA-nanoclay showed enhanced disease resistance (4% disease severity) and less sporulation (<1 × 10 ) compared with plants sprayed with dsRNA alone.

CONCLUSION

The use of nanoclay with multigene targeted dsRNA was assumed to be involved in effective delivery, protection and boosting the action of RNAi as a spray-induced gene silencing approach (SIGS). A significant reduction in growth, sporulation, disease severity and decreased gene expression authenticates the effects of SIGS on late blight progression. This study demonstrated as a proof of concept the dsRNA-nanoclay SIGS approach, which could be used as an alternative to chemical fungicides and transgenic approaches to develop an environmentally friendly novel plant protection strategy for late blight. © 2022 Society of Chemical Industry.

摘要

背景

卵菌纲致病疫霉是一种晚疫病的致病生物。它在新的杀菌剂分子释放后的几年内迅速进化和适应宿主背景,这很可能是由于其基因组中转座元件的优势。杀菌剂的频繁使用引起了环境问题。在这里,我们开发了基于靶标特异性 RNA 干扰 (RNAi) 的分子,以及纳米粘土载体,当喷洒在植物上时,能够有效地减少晚疫病的感染。

结果

根据我们自己的微阵列数据,针对孢子形成、早期感染和代谢途径阶段的特有基因。我们使用纳米粘土作为山梨醇脱氢酶、热休克蛋白 90、翻译延伸因子 1-α、磷脂酶-D 样 3 和糖基磷脂酰肌醇锚定酸性丝氨酸-苏氨酸丰富的 HAM34 样蛋白双链 (ds)RNA 的载体,通过培养生物测定、离体叶片测定和喷雾方法进行评估,结果显示生长、孢子形成和症状表达减少。与单独喷洒 dsRNA 的植物相比,喷洒多基因靶向 dsRNA-纳米粘土的植物表现出增强的抗病性 (4%的疾病严重程度) 和较少的孢子形成 (<1×10 )。

结论

纳米粘土与多基因靶向 dsRNA 的联合使用被认为可以有效传递、保护和增强 RNAi 作为喷雾诱导基因沉默 (SIGS) 的作用。生长、孢子形成、疾病严重程度的显著降低以及基因表达的降低证实了 SIGS 对晚疫病进展的影响。本研究证明了 dsRNA-纳米粘土 SIGS 方法作为一种替代化学杀菌剂和转基因方法的概念验证,可用于开发一种环境友好的新型植物保护策略来防治晚疫病。 © 2022 化学工业学会。

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