摘要
【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q PCR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。
【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.
作者
杨行洲
崔苗苗
魏利辉
顾爱国
李东霞
乐秀虎
冯辉
YANG Xingzhou;CUI Miaomiao;WEI Lihui;GU Aiguo;LI Dongxia;LE Xiuhu;FENG Hui(School of Landscape and Ecological Engineering,Hebei University of Engineering,Handan 056038,China;Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Jiangsu Product Quality Testing&Inspection Institute,Nanjing 210007,China)
出处
《中国水稻科学》
北大核心
2025年第5期703-710,共8页
Chinese Journal of Rice Science
基金
国家自然科学基金资助项目(31871943)
江苏省农业自主创新资金资助项目[cx(22)3017]
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]。
