基于 RNA-Seq 转录组分析鉴定稗草中甲磺隆非靶标位点抗性相关候选基因
RNA-Seq transcriptome analysis to identify candidate genes involved in non-target site-based mesosulfuron-methyl resistance in Beckmannia syzigachne.
机构信息
Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
出版信息
Pestic Biochem Physiol. 2021 Jan;171:104738. doi: 10.1016/j.pestbp.2020.104738. Epub 2020 Oct 24.
American sloughgrass (Beckmannia syzigachne Steud.) has become a dominant weed in fields with rice-wheat rotation. Moreover, herbicide resistance has rendered weed control difficult. We identified a biotype showing resistance to ALS inhibitor mesosulfuron-methyl with a resistant index 3.3, but without any ALS mutation. This study aims to identify and confirm the factors associated with non-target site resistance of this biotype to mesosulfuron-methyl using RNA-Seq. 118,111 unigenes were assembled, and 50.9% of these were annotated across seven databases. Eleven contigs related to metabolic resistance were identified based on differential expression via RNA-Seq which include a novel resistance-related transcription factor (MYC3) and two disease resistance proteins were also identified (At1g58602 and At1g15890). Fold changes in expression of these genes in comparison M-R vs. M-S ranged from 3.9 to 11.6, as confirmed by qPCR. The expression of a contig annotated as cytochrome P450 (CYP86B1) in resistant individuals was over 3 times higher than that in sensitive individuals at 0-72 h after mesosulfuron-methyl treatment. A similar trend was noted for three other genes annotated as glutathione S-transferase (GST), namely GST-T3, GST-U6, and GST-U14; the expression of GST-U6 in resistant individuals was up to 142.3 times higher than that in sensitive individuals at 24 h after mesosulfuron-methyl treatment. In addition, GST activity in resistant individuals was 2.1 to 5.3 times higher than that in sensitive individuals. The GR of resistant biotype decreased from 24.4 to 11.3 g a.i. ha after P450 inhibitor malathion treatment. This study identified a cytochrome P450 gene CYP86B1 and three GST genes GST-T3, GST-U6, and GST-U14 that have higher expression in mesosulfuron-methyl resistant B. syzigachne, suggesting that both P450- and GST-based activities could be involved in resistance.
美国黑麦草（Beckmannia syzigachne Steud.）已成为稻麦轮作田中的主要杂草。此外，抗除草剂性使得杂草防治变得困难。我们发现了一个对 ALS 抑制剂甲磺隆甲酯具有抗性的生物型，其抗性指数为 3.3，但没有任何 ALS 突变。本研究旨在使用 RNA-Seq 鉴定和确认该生物型对甲磺隆甲酯的非靶标部位抗性相关因素。共组装了 118111 个 unigene，其中 50.9%在七个数据库中进行了注释。通过 RNA-Seq 检测到 11 个与代谢抗性相关的基因差异表达的基因，包括一个新的抗性相关转录因子（MYC3）和两个抗病蛋白（At1g58602 和 At1g15890）。qPCR 验证了这些基因在 M-R 与 M-S 相比的表达倍数变化范围为 3.9 至 11.6。在甲磺隆甲酯处理后 0-72 小时，抗性个体中注释为细胞色素 P450（CYP86B1）的基因的表达水平比敏感个体高出 3 倍以上。在其他三个注释为谷胱甘肽 S-转移酶（GST）的基因（GST-T3、GST-U6 和 GST-U14）中也观察到类似的趋势；在甲磺隆甲酯处理后 24 小时，抗性个体中 GST-U6 的表达水平比敏感个体高出 142.3 倍。此外，抗性个体中的 GST 活性比敏感个体高 2.1 至 5.3 倍。在 P450 抑制剂马拉硫磷处理后，抗性生物型的 GR 从 24.4 降至 11.3 g a.i. ha。本研究鉴定出一个细胞色素 P450 基因 CYP86B1 和三个 GST 基因 GST-T3、GST-U6 和 GST-U14，它们在甲磺隆甲酯抗性 B. syzigachne 中的表达水平更高，这表明 P450 和 GST 活性都可能参与了抗性。
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