外源miR3979处理水稻对拟禾本科根结线虫趋性、侵染和发育的影响
收稿日期: 2024-09-18
修回日期: 2024-11-27
网络出版日期: 2025-09-10
基金资助
国家自然科学基金资助项目(31871943);河北省研究生创新资助项目(CXZZBS2024158);邯郸市可续技术研究与发展项目(21422012329)
Effects of Exogenous miR3979 on Chemotaxis, Infection and Development of Root Knot Nematode Meloidogyne graminicola in Rice
Received date: 2024-09-18
Revised date: 2024-11-27
Online published: 2025-09-10
【目的】miR3979-3p参与水稻响应多种生物和非生物胁迫，在拟禾本科根结线虫(Meloidogyne graminicola)侵染的根组织中下调表达，因此探究miR3979-3p对根结线虫与水稻互作中的作用具有重要意义。【方法】利用人工合成的双链miR3979(ds-miR3979)浸泡水稻根组织，采用qPCR测量水稻内源miR3979-3p的表达水平，通过平板接种和组织染色观察二龄幼虫的趋性和侵染能力，结合盆钵接种评价ds-miR3979处理对水稻根结产生和线虫发育的影响。【结果】100~800 nmol/L ds-miR3979处理12~24 h对二龄幼虫的运动活性无显著影响；与ddH2O处理的对照相比，400 nmol/L ds-miR3979浸泡根系12~24 h显著提高了立针期和三叶期水稻根系和叶片miR3979-3p的表达水平；连续显微观察2~8 h，发现二龄幼虫向水稻根尖移动趋性降低，集聚在根尖周围的线虫数量显著减少；同时，平板接种1~3 d侵入根内的线虫数量减少，盆钵接种7~21 d根系上的根结数目、根内雌虫数量和占比显著下降，且雌虫的发育进程延缓。【结论】人工合成的ds-miR3979可被水稻吸收并上调内源miR3979-3p表达，进而影响根结线虫的侵染和雌虫发育。
杨行洲, 崔苗苗, 魏利辉, 顾爱国, 李东霞, 乐秀虎, 冯辉 . 外源miR3979处理水稻对拟禾本科根结线虫趋性、侵染和发育的影响[J]. 中国水稻科学, 2025 , 39(5) : 703 -710 . DOI: 10.16819/j.1001-7216.2025.240906
【Objective】miR3979-3p is involved in rice responses to multiple biotic and abiotic stresses. Its expression is downregulated in root tissues infected by the root-knot nematode Meloidogyne graminicola. Therefore, it is important to investigate the role of miR3979-3p in the interaction between M. graminicola and rice.【Method】Rice roots were treated with artificially synthesized double-stranded miR3979 (ds-miR3979) via root soaking. Endogenous miR3979-3p expression levels were quantified using qPCR. Chemotaxis and infectivity of second-stage juveniles (J2s) were assessed through plate inoculation and tissue staining. The effects of ds-miR3979 treatment on root gall formation and nematode development were evaluated using pot inoculation assays.【Result】Treatment with 100-800 nmol/L ds-miR3979 for 12-24 h had no significant effect on J2 motility. Compared with ddH₂O-treated controls, root soaking with 400 nmol/L ds-miR3979 for 12-24 h significantly upregulated miR3979-3p expression in both roots and leaves of rice seedlings at the seedling emergence and three-leaf stages. Continuous microscopic observation revealed reduced chemotactic movement of J2s toward root tips within 2-8 h, with significantly fewer nematodes aggregating around root tips. Plate inoculation showed a decrease in the number of nematodes penetrating roots at 1-3 days post-inoculation (dpi). Pot inoculation assays demonstrated significant reductions in root gall number, number of females within roots, and proportion of females at 7-21 dpi, along with delayed female development.【Conclusion】Artificially synthesized ds-miR3979 can be taken up by rice plants, upregulating endogenous miR3979-3p expression, thereby inhibiting M. graminicola infection and female development.
Key words： small RNA; miR3979-3p; rice; Meloidogyne graminicola; infection; development
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