【摘要】 【目的】对棉花D基因组中Bet v 1基因家族进行分析,比较其在不同抗性棉种间的表达模式差异,为深入研究Bet v 1基因在棉花抗黄萎病中的作用提供理论依据。【方法】通过生物信息学方法对D基因组中Bet v 1基因进行鉴定。通过雷蒙德氏棉(Gossypium raimondii,D5)、三裂棉(G. trilobum,D8)和瑟伯氏棉(G.thurberi,D1)转录组和实时荧光定量聚合酶链式反应(qRT-PCR)分析Bet v 1基因在黄萎病菌处理下的表达模式。利用病毒诱导的基因沉默技术(VIGS)对Bet v 1基因进行功能鉴定。【结果】[棉花D基因组中包含59个成员,其中57个基因带有内含子,分布于8条染色体上,多数为亲水性蛋白并定位于细胞质。]黄萎病菌胁迫条件下3个野生棉种的Bet v 1基因表达量与其抗病水平一致。将不同表达水平的基因分为3组,其中第3组基因响应黄萎病菌侵染并在抗病棉种瑟伯氏棉中高表达,表明该组基因可能与黄萎病胁迫应答反应有关。从中筛选出高水平表达的Bet v 1基因,在陆地棉中沉默相应的同源基因,棉株感病加重,揭示该基因在棉花抵御黄萎病菌侵染过程中起正调控作用。【结论】响应黄萎病菌胁迫的Bet v 1基因在棉花抗黄萎病复杂的生物过程中至关重要。本研究结果为棉花Bet v 1家族基因的深入研究提供依据,为进一步解析棉花Bet v 1基因的功能奠定基础。更多还原

【Abstract】 [Objective] By analyzing the major birch pollen allergen Bet v 1 gene family in D genomes of cotton and comparing the expression patterns of three diploid D-genome cotton varieties with different Verticillium wilt resistance levels, we aimed to provide a theoretical basis for further studies on the role of Bet v 1 genes in cotton resistant to Verticillium wilt. [Method] The Bet v 1 genes were identified, and a bioinformatics analysis of the physicochemical properties of their encoded sequences in Gossypium raimondii（D₅） was performed. The transcriptome sequencing and quantitative real time-PCR of G. raimondii（D₅）,Gossypium trilobum（D₈） and Gossypium thurberi（D₁） were used to verify the expression patterns of Bet v 1 genes under Verticillium dahlia infection stress. Bet v 1 genes were silenced by virus-induced gene silencing in G. hirsutum to identify their functions. [Result] The D genome of cotton contains 59 members, 58 of which have introns and are distributed on eight chromosomes, and most encode hydrophilic proteins that localize to the cytoplasm. The expression levels of Bet v 1 genes in three wild cotton species having D genomes after being inoculated with V. dahliae were consistent with their disease resistance levels. The genes were separated into three groups based on their expression levels. Genes of Group 3 responded to V. dahliae infection and were highly expressed in disease-resistant cotton species G. thurberi. This indicated that Group 3 genes may be involved in the immune response of Verticillium wilt. A gene with a high expression level was screened out of Group 3. A corresponding homologous gene was silenced in G. hirsutum by virus-induced gene silencing, and gene-silenced plants were more susceptible to V. dahliae, indicating that the gene plays a positive regulatory role in the progress of Verticillium wilt resistance in cotton. [Conclusion] The Bet v 1 genes act in response to V. dahliae infection and are critical in cotton resistance to Verticillium wilt. The information obtained provides a basis for further studies of the cotton Bet v 1 family genes and their functions.更多还原