稻曲毒素A的相对含量分析及其与致病力的相关性

doi:10.3864/j.issn.0578-1752.2012.22.019

摘要/Abstract

摘要： 【目的】明确稻曲菌粗毒素的主要活性组分；比较不同地区稻曲球的稻曲毒素A含量；研究不同稻曲菌菌株产毒素A量与致病力的相关性。【方法】分别用丙酮、甲醇、水等不同极性溶剂从稻曲球中萃取制备粗毒素，并用小麦胚根抑制法检测其活性；高效液相色谱（HPLC）检测采集自全国224份稻曲球样品的毒素A含量，紫外扫描及液相色谱-质谱（LC-MS）验证稻曲毒素A；用人工接种的方法，测定52个稻曲菌株对感病品种两优培九的致病力；应用SPSS统计软件对稻曲菌产毒素A量与致病力进行相关性分析。【结果】用水和甲醇萃取得到的粗毒素活性较高，丙酮最低；质谱测定组分Ⅱ的分子量为673.9，结合紫外吸收光谱初步证明粗毒素的主要活性成分是稻曲毒素A。采集自不同地区的224份稻曲球样品的毒素A含量有明显差异。不同稻曲菌株的产毒素A量与致病力相关性不显著（相关系数R=0.222，显著性概率P=0.114）。【结论】稻曲球的活性成分中含有稻曲毒素A；不同地区稻曲球中的毒素A含量有显著差异；稻曲菌菌株的产毒素A量与其致病力没有显著相关性。

关键词: 稻曲菌 , 粗毒素 , 稻曲毒素A , 致病力

Abstract: 【Objective】The objective of this study is to define the active components of crude toxin produced by Ustilaginoidea virens, to analyze the ustiloxin A production of false smut balls in different areas, and to discover the correlation between pathogenicity and ustiloxin A production of the fungus.【Method】The crude toxin of U. virens was extracted from false smut balls by acetone, methanol and water, respectively, and the toxic effect was assayed and indicated by inhibition to wheat seed germination. The active components of the crude toxins were studied by liquid chromatography mass-spectrometry (LC-MS) and UV absorption spectra. Ustiloxin A production of 224 false smut balls collected from different areas in China was analyzed by high performance liquid chromatography (HPLC). The pathogenicity of 52 isolates of U. virens was examined through inoculating the susceptible rice cultivars, Liangyoupeijiu. The correlation between pathogenicity and ustiloxin A production of the fungus was analyzed by using SPSS software.【Result】The crude toxin extracted with water and methanol had higher inhibition to wheat radicle growth with the inhibition rate of 59.6% and 52.9%, respectively, while the crude toxin extracted with acetone had lower inhibition. The molecular weight of the active ingredient Ⅱwas 673.9 by determination with LC-MS. Combined with the UV spectra assay, ingredient Ⅱcould be proved to be ustiloxin A. It was found that there was a significant difference in the ustiloxin A production among false smut balls collected from different areas. However, correlation analysis indicated that no significant correlation between pathogenicity of the fungus and its ustiloxin A production（R=0.222, P=0.114）.【Conclusion】Water extract of false smut balls caused by U. virens contained ustiloxin A. And a significant difference in ustiloxin A production was found in false smut balls collected from different areas. There may be no association between pathogenicity and ustiloxin A production of the fungus.

Key words: Ustilaginoidea virens , crude toxin , ustiloxin A , pathogenicity

尹小乐, 陈志谊, 刘永锋, 于俊杰, 李燕, 俞咪娜. 稻曲毒素A的相对含量分析及其与致病力的相关性[J]. 中国农业科学, 2012, 45(22): 4720-4727.

YIN Xiao-Le, CHEN Zhi-Yi, LIU Yong-Feng, YU Jun-Jie, LI Yan, YU Mi-Na. Detection of the Relative Content of Ustiloxin A in Rice False Smut Balls and Correlation Analysis Between Pathogenicity and Ustiloxin A Production of Ustilaginoidea virens[J]. Scientia Agricultura Sinica, 2012, 45(22): 4720-4727.

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参考文献

[1]Shibata S, Ohta A, Ogihara Y. Metabolic products of fungi. XXI. On ustilaginoidins.1. the reactions of ustilaginoidin A. Chemical & Pharmaceutical Bulletin, 1963, 11: 1174-1178.

[2]Shibata S, Ogihara Y, Ohta A. Metabolic products of fungi. XXII. On ustilaginoidins.2. the structures of ustilaginoidin A. Chemical & Pharmaceutical Bulletin, 1963, 11: 1179-1182.

[3]Shibata S, Ogihara Y. Metabolic products of fungi. XXIII. On ustilaginoidins.3. the structure of ustilaginoidins B and C. Chemical & Pharmaceutical Bulletin, 1963, 11: 1576-1578.

[4]Koiso Y, Natori M, Iwasaki S. Ustiloxin: a phytotoxin and a mycotoxin from false smut balls on rice panicles. Tetrahedron Letters, 1992, 33(29): 4157-4160.

[5]Koiso Y, Li Y, Iwasaki S, Hanaoka K, Kobayashi T, Sonoda R, Fujita Y, Yaegashi H, Sato Z. Ustiloxins, antimitotic cyclic peptides from false smut balls on rice panicles caused by Ustilaginoidea virens. The Journal of Antibiotics, 1994, 47(7): 765-773.

[6]Koiso Y, Morisaki N, Yamashita Y, Mitsui Y, Shirai R, Hashimoto Y, Iwasaki S. Isolation and structure of an antimitotic cyclic peptide, ustiloxin F: chemical interrelation with a homologous peptide, ustiloxin B. Chemlnform, 1998, 29(39): DOI: 10.1002/Chin. 199839219.

[7]王疏, 刘晓舟, 白元俊, 邵雪, 褚茗莉, 朱茂山. 稻曲病菌毒素研究初报. 辽宁农业科学, 1995(1): 52-54.

Wang S, Liu X Z, Bai Y J, Shao X, Zhu M L, Zhu M S. Preliminary research report on toxin produced by Ustilaginoidea virens. Liaoning Agricultural Sciences, 1995(1): 52-54. (in Chinese)

[8]陈美军, 胡东维, 徐颖. 稻曲毒素的活性测定、抗体制备与细胞定位. 实验生物学报, 2004, 37(4): 310-314.

Chen M J, Hu D W, Xu Y. Activity assay, antiserum localization preparation and cellular localization of ustiloxins. Acta Biologiae Experimentalis Sinica, 2004, 37(4): 310-314. (in Chinese)

[9]Miyazaki S, Matsumoto Y, Uchihara T, Morimoto K. High- performance liquid chromatographic determination of ustiloxin A in forage rice silage. The Journal of Veterinary Medical Science, 2009, 71(2): 239-241.

[10]Luduena R F, Roach M C, Prasad V, Banerjee M, Koiso Y, Li Y, Iwasaki S. Interaction of ustiloxin A with bovine brain tubulin. Biochemical Pharmacology, 1994, 47(9): 1593-1599.

[11]Li P, Evans C D, Wu Y, Cao B, Hamel E, Joullie M M. Evolution of the total syntheses of ustiloxin natural products and their analogues. Journal of the American Chemical Society, 2008, 130(7): 2351-2364.

[12]Cao B, Park H, Joullie M M. Total synthesis of ustiloxin D. Journal of the American Chemical Society, 2002, 124(4): 520-521.

[13]Hutton C A, White J M. Towards a total synthesis of ustiloxins A and B. stereocontrolled synthesis of (2S, 4S, 6S)-4-hydroxy-5- phenylsulfinylnorvaline. Tetrahedron Letters, 1997, 38(9): 1643-1646.

[14]Sawayama A M, Tanaka H, Wandless T J. Total synthesis of ustiloxin D and considerations on the origin of selectivity of the asymmetric allylic alkylation. The Journal of Organic Chemistry, 2004, 69(25): 8810-8820.

[15]张君成, 陈志谊, 张炳欣, 刘永锋, 陆凡. 稻曲病的接种技术研究. 植物病理学报, 2004, 34(5): 463-467.

Zhang J C, Chen Z Y, Zhang B X, Liu Y F, Lu F. Inoculation techniques used for inducing rice false smut efficiently. Acta Phytopathologica Sinica, 2004, 34(5): 463-467. (in Chinese)

[16]唐春生, 高家樟, 曹国平, 黄守行, 刘二明, 刘承龙. 稻曲病病情分级标准的研究和应用. 植物保护, 2001, 27(1): 18-21.

Tang C S, Gao J Z, Cao G P, Huang S X, Liu E M, Liu C L. Research and application of classification standard of rice false smut. Plant Protection, 2001, 27(1): 18-21. (in Chinese)

[17]康振生, 黄丽丽, Buchenauer H. 小麦穗组织中脱氧镰刀菌烯醇毒素的免疫细胞化学定位. 植物病理学报, 2004, 34(5): 419-424.

Kang Z S, Huang L L, Buchenauer H. Immunocytochemical localization of deoxynivalenol in infected wheat spikes by Fusarium graminearum. Acta Phytopathologica Sinica, 2004, 34(5): 419-424. (in Chinese)

[18]齐俊生, 李怀方. 一种检测棉花黄萎菌毒素致萎性的新方法—叶片针刺涂抹法. 棉花学报, 2006, 18(4): 228-232.

Qi J S, Li H F. A new detection method of wilting induction by phytotoxin from V. dahliae on cotton through leaf pricking and spreading. Cotton Science, 2006, 18(4): 228-232. (in Chinese)

[19]杨艳丽, 肖浪涛, 胡先奇. 马铃薯晚疫病菌毒素与寄主品种抗性关系研究. 中国农业科学, 2009, 42(6): 2202-2210.

Yang Y L, Xiao L T, Hu X Q. Study on the relationship between the toxin of Phytophthora infestans and resistance of potato. Scientia Agricultura Sinica, 2009, 42(6): 2202-2210. (in Chinese)

[20]俞刚, 陈利锋, 姚红燕, 柴一秋. 脱氧雪腐镰刀菌烯醇在小麦赤霉病病程中的作用. 植物病理学报, 2003, 33(1): 40-43.

Yu F, Chen L F, Yao H Y, Chai Y Q. Role of deoxynivalenol in pathogenesis of wheat head scab. Acta Phytopathologica Sinica, 2003, 33(1): 40-43. (in Chinese)

[21]桑茜, 袁虹霞, 王振跃, 李洪连, 汪敏. 河南省不同地区棉花黄萎病菌分离物致病性及其毒素致萎活性测定. 棉花学报, 2010, 22(4): 333-338.

Sang X, Yuan H X, Wang Z Y, Li H L, Wang M. Pathogenicity of Verticillium dahliae cotton isolates in Henan province and the wilting capability of their crude toxins. Cotton Science, 2010, 22(4): 333-338. (in Chinese)

[22]陈丽, 胡东维, 陈美军, 张敬泽. 稻曲球及稻曲病菌菌落微结构的SEM观察. 菌物学报, 2007, 26(1): 89-96.

Chen L, Hu D W, Chen M J, Zhang J Z. SEM observation of rice false smut balls and colony microstructure of Ustilaginoidea virens. Mycosystema, 2007, 26(1): 89-96. (in Chinese)