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水稻抗褐飞虱基因QBPH1和QBPH4的精细定位与聚合效应分析

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[目的]利用遗传群体发掘新的水稻抗褐飞虱QTL,并评估其聚合效应,为抗虫育种提供重要的遗传材料和基因资源。[方法]利用日本晴(感虫型)和佳辐占(抗虫型)的重组自交系(RILs)群体,结合苗期抗性评价和极端混池测序法(BSA-seq)定位抗褐飞虱QTL。进一步通过QTL精细定位、候选基因鉴定、聚合效应分析及混池转录组测序(BSR-seq),揭示QTL介导的抗性生理和分子机制。[结果]BSA-seq分析,在水稻第1(30—32 Mb区段)和第4(5—7 Mb区段)染色体上定位到2个主效抗性QTL,分别命名为QBPH1和QBPH4。通过区间连锁的分子标记,验证了这些QTL的真实性。QBPH4与已克隆的BPH3和BPH15位于同一位置,而QBPH1为一个新发现的QTL。通过加密标记和重组单株分析,将QBPH1的位置精确到30。61—30。65 Mb。在此区域内,Os01g53294和Os01g53330分别编码呼吸爆发氧化酶蛋白B和花青素5,3-O-葡萄糖基转移酶,被认为是QBPH1的可靠候选基因。此外,通过对比QBPH1和QBPH4单基因及其聚合家系在苗期的抗性、褐飞虱蜜露面积和重量、虫增重、致死率、偏好性等指标,发现双基因聚合家系的抗性未显著增强。QBPH1和QBPH4均介导了抗生性和趋避性的生理机制,但其效应存在差异。BSR-seq分析揭示了QBPH1的不同等位基因型在转录调控、蛋白质氨基酸磷酸化和氧化还原过程中的差异表达基因(DEGs)显著富集。此外,与茉莉酸(jasmonic acid,JA)合成和信号传导路径相关的基因在抗虫材料中得到显著上调,并通过RT-qPCR试验得到了验证。[结论]成功在水稻第1染色体上鉴定了一个新的抗褐飞虱QTL——QBPH1。该QTL介导了水稻对褐飞虱的抗生性和趋避性,但与另一QTL的聚合效应并不显著。QBPH1可能通过参与JA通路来介导对褐飞虱的防御机制。在此基础上,Os01g53294和Os01g53330被确定为QBPH1的可靠候选基因。
Fine Mapping and Analysis of Pyramiding Effects of Rice Brown Planthopper Resistance Genes QBPH1 and QBPH4
[Objective]This study aims to discover new quantitative trait loci (QTLs) for resistance to the brown planthopper (BPH) in rice by utilizing a genetic population and to assess the pyramiding effects of these QTLs,thereby providing crucial genetic materials and resources for breeding insect-resistant rice.[Method]A recombinant inbred line (RIL) population derived from a cross between susceptible NPB and resistant Jiafuzhan was used,combined with seedling stage resistance evaluation and extreme mixed pool sequencing (BSA-seq) to map BPH resistance QTLs.Further,fine mapping,candidate gene identification,pyramiding effect analysis,and mixed-pool transcriptome sequencing (BSR-seq) were employed to elucidate the physiological and molecular mechanisms mediated by these QTLs.[Result]BSA-seq identified two major resistance QTLs on chromosomes 1 (30-32 Mb) and 4 (5-7 Mb),named QBPH1 and QBPH4,respectively.Interval-linked molecular markers confirmed the authenticity of these QTLs.QBPH4 coincides with previously cloned BPH3 and BPH15,while QBPH1 represents a newly discovered QTL.High-density markers and recombinant analysis further narrowed the QBPH1 region to 30.61-30.65 Mb.This analysis identified Os01g53294 and Os01g53330 as reliable candidate genes,which code for a respiratory burst oxidase protein B and an anthocyanin 5,3-O-glucosyltransferase,respectively.In evaluations of seedling resistance,brown planthopper honeydew area and weight,insect weight gain,lethality,and preference,comparisons between QBPH1 and QBPH4 single genes and their polymerized lines revealed no significant enhancement in resistance.Both QBPH1 and QBPH4 mediated antibiosis and antixenosis mechanisms with varying effects.BSR-seq analysis highlighted significant enrichment of differentially expressed genes (DEGs) involved in transcriptional regulation,protein phosphorylation,and redox processes among different QBPH1 alleles.Additionally,genes associated with jasmonic acid (JA) synthesis and signaling pathways were significantly upregulated in resistant materials,confirmed by RT-qPCR experiments.[Conclusion]A novel BPH resistance QTL,QBPH1,was successfully identified on chromosome 1 of rice.QBPH1 mediated both antibiosis and avoidance against BPH,though its aggregation effect with another QTL was not significant.QBPH1 may mediate defense mechanisms against BPH through involvement in the JA pathway.Based on this,Os01g53294 and Os01g53330 have been validated as reliable candidate genes for QBPH1.

ricebrown planthopper resistanceQTL mappinggene pyramidingRNA-seq

熊尚烨、张翔、梁宝慧、叶仰东、李浴阳、朱晓、朱志鸿、官华忠、张帅、吴建国、胡杰

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福建农林大学植物保护学院/闽台作物有害生物生态防控国家重点实验室/媒介病毒研究中心,福州 350002

福建农林大学农学院/福建省作物设计育种重点实验室,福州 350002

福建农林大学未来技术学院/遗传改良研究中心,福州 350002

水稻 褐飞虱抗性 QTL定位 基因聚合 转录组测序

2024

中国农业科学
中国农业科学院

中国农业科学

CSTPCD北大核心
影响因子:1.899
ISSN:0578-1752
年,卷(期):2024.57(23)