为揭示稻曲病菌厚垣孢子壁与厚垣孢子内源性休眠的关系,以典型的休眠型(黑色)和非休眠型(黄色)稻曲病菌厚垣孢子为材料,用液氮研磨,超声破壁后,经复合酶热水浸提Sevag法提取胞壁多糖,采用紫外光谱、红外光谱、刚果红实验、气相色谱分析其多糖的结构和组分。结果表明,黑色、黄色厚垣孢子胞壁多糖均含β型吡喃糖苷键,具有三股螺旋构象,为主要由葡萄糖、半乳糖、甘露糖3种单糖组成的杂多糖,黄色厚垣孢子中含有少量的木糖;黑色厚垣孢子壁中葡萄糖、半乳糖和甘露糖的摩尔百分比为58.23∶25.32∶16.46,黄色厚垣孢子壁中葡萄糖、半乳糖、甘露糖和木糖的摩尔百分比为3548∶30.58∶24.46∶9.45。这两种厚垣孢子壁多糖的组成和比例有明显差异,可能与厚垣孢子休眠有关。
To reveal the relationship between endogenous dormancy mechanism of chlamydospores of U.virens and their walls, polysaccharide from cell walls of the dormant (black) and nondormant (yellow) chlamydospores was obtained by complex enzymehot water extractionSevag after the chlamydospores wall breaking through liquid nitrogen grinding plus ultrasonic crushing technique, and structure and composition of cell wall polysaccharide were analyzed by ultraviolet absorption spectrum (UV spectrum),infrared absorption spectrum (IR spectrum), Congo red test, and gas chromatography (GC). The results showed that the polysaccharide from two kinds of the chlamydospores wall was acidic heteropolysaccharide and mainly consisted of the three monosaccharides, glucose, galactose and mannose, but the yellow chlamydospores wall still contained a small amount of xylose. Further, polysaccharide in the black chlamydospores wall, was composed of glucose, galactose, mannose with a molar percentage ratio of 58.23∶25.32∶16.46,while the yellow chlamydospore wall was composed of glucose, galactose, mannose and xylose, with a molar percentage ratio of 35.48∶30.58∶24.46∶9.45. The composition and composition ratios of monosaccharides in polysaccharide from the dormant and nondormant chlamydospore walls obviously varied, which may be closely related to chlamydospore dormancy.
\[1\]Ashizawa T, Takahashi M, Moriwaki J, et al. A refined inoculation method to evaluate false smut resistance in rice. J Gener Plant, 2011, 77: 1016.
\[2\]王国良. 稻曲病菌厚垣孢子浸染时期和侵染途径的研究. 植物保护学报, 1992, 19(2): 97100.
\[3\]陆凡, 陈志谊, 陈琉苓. 稻曲病菌的生物学特性及其侵染循环中某些未确定要点的研究. 江苏农业学报, 1996, 12(4): 3540.
\[4\]王疏, 白元俊, 周永力, 等. 稻曲病菌的病原学. 植物病理学报, 1998, 28(1): 1924.
\[5\]燕玮婷, 刘二明, 邓林伟,等. 稻曲病菌不同颜色厚垣孢子超微结构比较. 植物病理学报, 2010, 40(5): 538542.
\[6\]周鹏, 谢明勇, 付博强. 多糖的结构研究. 南昌大学学报:理科版, 2001, 25(2): 197202.
\[7\]Srivastava R, Dinesh K. Bioactive polysaccharides from plants. Phytochemitry, 1989, 28(11): 28772883.
\[8\]燕玮婷, 刘二明, 邓林伟, 等. 稻曲病菌厚垣孢子不同破壁方法的比较. 微生物学杂志, 2010, 30(2): 1417.
\[9\]王娜, 任佐华, 邓林伟, 等. 稻曲病菌厚垣孢子壁多糖的提取方法优选. 微生物学通报, 2011, 38(9): 14121417.
\[10\]何晋浙, 邵平, 孟祥河, 等. 灵芝多糖的结构特征分析. 分析化学, 2010, 38(3) : 372376.
\[11\]Barker P E, Hatt B W, Holding S R. Dissolution of silaceous chromatogiaphic packings of dextran at enhanced flowrates. J Chrom,1981, 206: 2734.
\[12\]Edge C J, Rademacher T W, Wormald M R, et al. Fast sequencing of oligosaccharides: The reagentarray analysis method. PNAS, 1992, 89(4): 63386342.
\[13\]Prieto A, Leal J A, GiménezAbián M I, et al. Isolation and structural determination of a unique polysaccharide containing mannofuranose from the cell wall of the fungue Acrospermum compressum. J Glycoc, 2007, 24: 421429.
\[14\]Leal J A, GiménezAbián M I, Canales A′, et al. Cell wall polysaccharides isolated from the fungus Neotestudina rosatii, one of the etiologic agents of mycetoma. J Glycocon, 2009,26:10471054.
\[15\]陆家云. 植物病原真菌学. 北京: 中国农业出版社, 2011: 2021.
\[16\]Deacon J W. Introduction to Morden Mycology.2nd. Oxford:Blackwell Science:1984.
\[17\]ELMougtth A A. Effect of benomyl on the growth and lipid composition of Trichoderma koningii. Folia Microbiol,1999, 44(1): 4144.
\[18\]刘志恒. 现代微生物学. 2版. 北京: 科学出版社, 2008.