| 冯雪琪 1, 2, 马如清 1, 2, 杨文娟 1, 2, 李 妍 1, 2, 汪军成 1, 2, 孟亚雄 1, 2, 姚立蓉 1, 2, 王化俊 1, 2, 司二静 1, 2*, 侯 璐 1, 3*.大麦条纹病菌Pg00232基因致病功能研究[J].植物保护,2026,52(4):37-45. |
| 大麦条纹病菌Pg00232基因致病功能研究 |
| Pathogenic function analysis of the Pg00232 gene in Pyrenophora graminea |
| 投稿时间：2025-06-06 修订日期：2025-09-27 |
| DOI：10.16688/j.zwbh.2025268 |
| 中文关键词: 大麦条纹病菌 亚细胞定位 RNAi 致病性 |
| 英文关键词:Pyrenophora graminea subcellular localization RNA interference pathogenicity |
| 基金项目:国家自然科学基金(32160496, 32160647, 32460517);国家大麦青稞产业技术体系(CARS-05-02A-02);甘肃省教育厅产业支撑计划(2021CYZC-12);甘肃省现代寒旱特色农业麦类产业技术体系(GSARS-070);甘肃农业大学大学生创新创业训练项目(202501068, 202501036) |
| 作者 | 单位 | E-mail | | 冯雪琪 1, 2, 马如清 1, 2, 杨文娟 1, 2, 李 妍 1, 2, 汪军成 1, 2, 孟亚雄 1, 2, 姚立蓉 1, 2, 王化俊 1, 2, 司二静 1, 2*, 侯 璐 1, 3* | 1. 甘肃农业大学农学院, 兰州 730070
2. 干旱生境作物学国家重点实验室, 甘肃省作物遗传改良与种质创新重点实验室, 兰州 730070
3. 青海大学农林科学院, 青海 810016 | 司二静siej@gsau.edu.cn;侯璐mantou428@163.com |
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| 中文摘要: |
| 大麦条纹病(barley leaf stripe)由大麦条纹病菌Pyrenophora graminea引起, 主要通过带菌种子传播。本研究采用生物信息学、RNAi技术初步研究了Pg00232基因对大麦条纹病菌致病性的调控作用。结果表明, Pg00232基因全长624 bp, 编码207个氨基酸, 其编码蛋白具有信号肽、为不稳定亲水性蛋白。亚细胞定位显示该蛋白定位于寄主细胞核与细胞膜。采用PEG介导法转化原生质体, 获得了2个RNAi干扰株Pg00232-RNAi-1和Pg00232-RNAi-2, RT-qPCR结果显示, 2个菌株中Pg00232的表达量较野生型菌株QWC分别下降了30.73%和48.92% (P<0.05), 且RNAi干扰株接种的大麦发病率较野生型菌株QWC分别显著降低了55.11%和59.69% (P<0.05), 生长速率较QWC分别降低了20.00%和24.44%(P＜0.05)。经正倒置一体显微镜观察发现, RNAi干扰株菌丝相互缠绕或出现不规则断裂。台盼蓝和DAB染色显示, RNAi干扰株侵染的大麦叶片内死亡细胞数量与活性氧积累水平均显著低于QWC。本研究结果表明, Pg00232不仅参与调控大麦条纹病菌的菌丝生长与发育, 而且在其致病过程中发挥关键作用, 为进一步解析大麦条纹病菌的致病机制提供了理论依据。 |
| 英文摘要: |
| Barley leaf stripe, caused by Pyrenophora graminea, is mainly transmitted through infected seeds. In this study, bioinformatics analysis and RNA interference (RNAi) technology were utilized to preliminarily investigate the regulatory role of the Pg00232 gene in the pathogenicity of P.graminea. The results showed that the full-length Pg00232 gene is 624 bp, encoding 207 amino acids. The encoded protein contains a signal peptide and is an unstable hydrophilic protein. Subcellular localization analysis revealed that the protein is localized in both the nucleus and the cell membrane of host. Two RNAi silencing strains, Pg00232-RNAi-1 and Pg00232-RNAi-2, were successfully generated via PEG-mediated protoplast transformation. RT-qPCR analysis showed that the expression levels of Pg00232 in the two strains decreased by 30.73% and 48.92%, respectively, compared with the wild-type strain QWC. Moreover, the disease incidence of barley infected by the RNAi silencing strains was significantly reduced by 55.11% and 59.69%, respectively, compared with QWC, and their growth rates decreased by 20.00% and 24.44%, respectively (P<0.05). Observation under an integrated upright and inverted microscope showed that the hyphae of the RNAi silencing strains were intertwined or exhibited irregular breakage. Trypan blue and DAB staining indicted that both the number of dead cells and the level of reactive oxygen species accumulation in barley leaves infected by the RNAi silencing strains were significantly reduced, in comparison with those leaves infected by QWC. These results indicate that Pg00232 not only regulates hyphal growth and development of P.graminea, but also plays a critical role in its pathogenesis, providing a theoretical basis for further dissecting the pathogenic mechanism of P.graminea. |
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