不同大豆与大豆花叶病毒组合中胼胝质的荧光标记及其功能初探
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国家自然科学基金;河北省自然科学基金


Study on callose fluorescence labeling and function in the different soybean-soybean mosaic virus combinations
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National Natural Science of China;The natural science foundation of Hebei Province

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    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,即品种的抗病性越强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,品种的抗病性越强就会在侵染点处越早的积累胼胝质,胼胝质的沉积与大豆抗病毒侵染密切相关。

    Abstract:

    The selected six varieties of soybean and four different strains of soybean mosaic virus(SMV) are divided into nine combinations that respond on different resistance levels. Through the symptom observation of inoculated leaves and the upper leaves, aniline blue staining followed by fluorescence microscopy, and pharmacological experiment we want to study the characteristics of callose (β- 1,3 - glucan ) accumulation in combinations of different resistance levels and the function of callosein in the process of soybean and SMV interaction. The results show that,in the earlier period of time after inoculation (6-72 h ), callose fluorescence in the mesophyll cells of different combinations, 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 callose fluorescence is closely related to the resistance levels.In another word, the variety more strongly resisting to SMV can earlier be found the callose fluorescen at the sites of inoculation. However, callose fluorescence has not been observed in the combination that responds in resistance level 5 . In addition, in the combinations responding in resistance level 0 and 1, necrosis in the upper leaves was observed and the viral RNA of specific coat protein(CP-RNA) was detected by RT-PCR analysis following pre-injection of 2-DDG ( an inhibitor of callose synthesis).. Therefore, the deposition of callose at the sites of inoculation can 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 be found the callose accumulation at the sites of inoculation .Callose accumulation relates closely to the resistance of soybean to SMV.

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吴思思,李文龙,肖东强,等.不同大豆与大豆花叶病毒组合中胼胝质的荧光标记及其功能初探[J].植物遗传资源学报,2013,14(1):134-142.

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  • 收稿日期:2012-01-03
  • 最后修改日期:2012-10-10
  • 录用日期:2012-10-27
  • 在线发布日期: 2012-12-27
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