大豆不同花叶病毒抗性品种胼胝质荧光标记初探  

Callose Fluorescence Labeling in the Different Soybean Varieties Resistant to Soybean Mosaic Virus

作　　者：吴思思[1] 李文龙[1] 肖东强[1] 侯春燕[1] 王冬梅[1] 

机构地区：[1]河北农业大学生命科学学院,保定071001

出　　处：《植物遗传资源学报》2013年第1期132-140,共9页Journal of Plant Genetic Resources

基　　金：国家自然科学基金项目(30971706);河北省自然科学基金项目(C2008000321)

摘　　要：选用6个大豆品种与4个不同的大豆花叶病毒株系,分别组成抗病级别不同的组合,通过对接种叶片与上位叶症状观察、苯胺蓝染色辅以荧光显微镜观察和药物学试验,探讨了不同抗病级别组合中胼胝质(即β-l,3-葡聚糖)积累的特点及其在大豆抵抗大豆花叶病毒侵染过程中的作用。试验结果表明,大豆接种病毒后,在抗病级别分别为0～3的各个组合的叶肉细胞中,在侵染早期(接种后6、72 h)不同的组合在不同时间点分别观察到了胼胝质荧光,且胼胝质荧光出现的时间与抗病级别密切相关,即抗病性越强的组合在侵染点处观察到胼胝质的时间越早;而在抗病级别为5的组合中一直未能观察到胼胝质荧光。另外,在抗病级别为0级和1级的各组合中给叶片预注射2-DDG(2-deoxy-D-glucose,一种胼胝质合成抑制剂)再接种病毒,在上位叶能观察到坏死斑的出现并且通过RT-PCR能够检测到大豆花叶病毒外壳蛋白基因。以上结果表明,大豆被大豆花叶病毒侵染后,抗病性越强的品种就会在侵染点处越早地积累胼胝质,胼胝质的沉积与大豆抗病毒侵染密切相关。The selected six varieties of soybean and four different strains of soybean mosaic virus （SMV） are di- vided into nine combinations that respond to different resistance levels. Through the symptom observation of inocula- ted leaves and the upper leaves, aniline blue staining followed by fluorescence microscopy, and pharmacological ex- periment, we wanted to study the characteristics of callose （ 13-1,3-glucan） accumulation in combinations of different resistance levels and the function of callose in the process of soybean and SMV interaction. The results showed that, in the earlier period of time after inoculation（6-72 h） , callose fluorescence in the mesophyll cells of different combi- nations,which resistance levels ranges from 0 to 3, were observed at different time depending on different resistant leveles of combinations and the emergence time of eallose fluorescence was closely related to the resistance levels. In another words,the variety was more strongly resisting to SMV and can earlier find the callose fluorescen at the sites of inoculation. However,callose fluorescence had not been observed in the combination that responded in resistance level 5. In addition,in the combinations responding in resistance level 0 and 1, necrosis in the upper leaves was ob- served and the viral RNA of specific coat protein（CP-RNA） was detected by RT-PCR analysis following pre-injec- tion of 2-DDG （ an inhibitor of callose synthesis）. Therefore, the deposition of callose at the sites of inoculation could limit the transportation of SMV in mesophyll cells,indicating that callose plays an important role in the resistance of soybean to the infection of SMV. All the results above indicated that, the variety more strongly resisting to SMV can earlier find the callose accumulation at the sites of inoculation . Callose accumulation related closely to the resist- ance of soybean to SMV.

关 键 词：大豆 大豆花叶病毒 胼胝质 

分 类 号：S565.1[农业科学—作物学]