玉蜀黍黑粉菌Bizl调控基因功能研究
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摘要
玉米作为重要的粮食作物、饲料作物及工业原料，在我国的农业生产和国民经济发展中占有重要的地位。玉米瘤黑粉病作为一种真菌病害，在世界各玉米主产区均有发生，影响玉米的产量和品质，因此如何防治玉米瘤黑粉病是一个极具现实意义的问题。近来的研究结果表明，玉蜀黍黑粉菌biz1基因在敲除后会导致侵染能力丧失，从而推断玉蜀黍黑粉菌的侵染能力与Biz1蛋白有关。在玉蜀黍黑粉菌侵染后的发育过程中，bip系列基因（受Biz1调控基因）的表达水平发生明显变化，故而推测这些基因可能与玉蜀黍黑粉菌的侵染能力有关。
本课题主要是通过对部分bip系列基因进行过表达和敲除，构建黑粉菌突变体菌株，然后利用所构建的突变体菌株进行玉米植株侵染实验并观测其发病情况从而检测基因功能。通过研究与玉蜀黍黑粉菌侵染能力相关的基因，为防治玉米瘤黑粉病以及进行玉米植株抗病品种的研究提供理论依据。
在实验过程中，我们发现黑粉菌中srp1（bip5）基因的缺失会导致受侵染的玉米植株发病程度加重，黑粉瘤的大小和数量与野生型相比均有明显的增长；而srp1基因过表达后会使受侵染的玉米植株的发病程度明显减弱，主要表现在黑粉瘤的大小和数量与野生型相比均有明显的减少。这些结果显示srp1基因的表达对玉蜀黍黑粉菌侵染后的的生长发育产生抑制作用。
Maize is an important grain crops, feed crops and raw materials for industry, whichholds the important status in our country’s agricultural production and the nationaleconomy development. Corn smut, one kind of worldwide plant disease, can affectthe maize yield and quality. Therefore it is very important to investigate this disease inmore details. Recent results indicate that the biz1gene deletion mutant can not infectthe corn any more. So we speculate the pathogenic development of Ustilago maydisafter plant penetration is regulated by Biz1. During the pathogenic growth process ofUstilago maydis, bip genes（Biz1induced protein）are up-regulated by the Biz1protein, therefore these genes may involve in the pathogenic development of Ustilagomaydis after plant infection.
In this work, some candidates of bip genes are selected for the over expression anddeletion to identify their functions by plant infection experiments. Symptoms aftermaize infection were compared during these mutant strains to find out the function ofthese genes. The results can provide the theory basis for corn smut prevention andcure.
The plant infection experiment showed that the maize had more severe incidenceafter infection by the srp1deletion mutants, the size and number of tumors had anobvious growth compared with the wild-type strains; the srp1over expressionmutants reduced the incidence, the size and number of tumors had an obviousdecrease compared with the wild-type strains. Taken together, the expression of srp1gene can suppress the pathogenic development of Ustilago maydis after plantinfection.
本课题主要是通过对部分bip系列基因进行过表达和敲除，构建黑粉菌突变体菌株，然后利用所构建的突变体菌株进行玉米植株侵染实验并观测其发病情况从而检测基因功能。通过研究与玉蜀黍黑粉菌侵染能力相关的基因，为防治玉米瘤黑粉病以及进行玉米植株抗病品种的研究提供理论依据。
在实验过程中，我们发现黑粉菌中srp1（bip5）基因的缺失会导致受侵染的玉米植株发病程度加重，黑粉瘤的大小和数量与野生型相比均有明显的增长；而srp1基因过表达后会使受侵染的玉米植株的发病程度明显减弱，主要表现在黑粉瘤的大小和数量与野生型相比均有明显的减少。这些结果显示srp1基因的表达对玉蜀黍黑粉菌侵染后的的生长发育产生抑制作用。
Maize is an important grain crops, feed crops and raw materials for industry, whichholds the important status in our country’s agricultural production and the nationaleconomy development. Corn smut, one kind of worldwide plant disease, can affectthe maize yield and quality. Therefore it is very important to investigate this disease inmore details. Recent results indicate that the biz1gene deletion mutant can not infectthe corn any more. So we speculate the pathogenic development of Ustilago maydisafter plant penetration is regulated by Biz1. During the pathogenic growth process ofUstilago maydis, bip genes（Biz1induced protein）are up-regulated by the Biz1protein, therefore these genes may involve in the pathogenic development of Ustilagomaydis after plant infection.
In this work, some candidates of bip genes are selected for the over expression anddeletion to identify their functions by plant infection experiments. Symptoms aftermaize infection were compared during these mutant strains to find out the function ofthese genes. The results can provide the theory basis for corn smut prevention andcure.
The plant infection experiment showed that the maize had more severe incidenceafter infection by the srp1deletion mutants, the size and number of tumors had anobvious growth compared with the wild-type strains; the srp1over expressionmutants reduced the incidence, the size and number of tumors had an obviousdecrease compared with the wild-type strains. Taken together, the expression of srp1gene can suppress the pathogenic development of Ustilago maydis after plantinfection.
引文
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[2]. Agrios, G.N. Plant diseases caused by fungi. In: Plant Pathology,4th edn. Agrios, G.N.(ed.).San Diego, CA: Academic Press, pp,1997,245–406.
[3]. Mendgen, K.&Hahn, M. Plant infection and the establishment of fungal biotrophy. TrendsPlant Sci.2002,7:352–356.
[4].阙友雄，宋弦弦，许莉萍，陈如凯.植物与病原真菌互作机制研究进展.生物技术通讯,2009:282-285.
[5]. Perkins, D.D. Biochemical mutants in the smut fungus, Ustilago maydis. Genetics,1949,34:607–626.
[6]. Christensen, J.J. Corn smut caused by Ustilago maydis. Monograph number2. AmericanPhytopathological Society, St. Paul, MN,1963.
[7]. Holliday, R.. Ustilago maydis. In: King, R.C.(Ed.), Handbook of Genetics, vol.1.Plenum Press, New York, pp,1974,575–595.
[8]. Martinez-Espinoza, A.D., Garcia-Pedrajas, M.D., Gold, S.E. The Ustilaginales as plant pestsand model systems. Fungal Genet. Biol,2002,35:1–20.
[9]. Holloman, W.K., Schirawski, J., Holliday, R. Towards understanding the extreme radiationresistance of Ustilago maydis. Trends Microbiol,2007,15:525–529.
[10].石太渊，孙大为，张锐,张华,姜福林.玉米黑粉菌的加工利用分析.农产品加工·学刊,2006,(9):28-32.
[11].张雪君，黄丽敏，胡海军.玉米黑粉病及其综合防治.中国种业,2001,(1):38.
[12].刘正坪，赵占军.玉米瘤黑粉病菌冬孢子生物学特性研究[J].现代化农业,1998,(1):12-13.
[13]. Banuett, F. Genetics of Ustilago maydis, a fungal pathogen that induces tumors in maize.Annu. Rev. Genet,1995,29:179–208.
[14]. CHRISTOPH W. BASSE AND GERO STEINBERG. Ustilago maydis, model system foranalysis of the molecular basis of fungal pathogenicity. MOLECULAR PLANT PATHOLOGY,2004,5(2):83–92.
[15].刘素敏.玉米瘤黑粉病的发生规律及防治方法.植物保护,2008,(23):38-39.
[16].韩睿，邓灵福，黎晨，张青叶，张劼，高强，熊丽，万坚，刘德立.玉米黑粉菌cyp51基因结构分析与克隆表达.生物工程学报,2008,24(10):1747–1753.
[17]. Madden, L.V. and Wheelis, M. The threat of plant pathogens as weapons against U.S. crops.Annu. Rev. Phytopathol,2003,41:155–176.
[18]. Michael E. Donaldson, Barry J. Saville. Bioinformatic identification of Ustilago maydismeiosis genes. Fungal Genetics and Biology,2008,45: S47–S53.
[19].王辉，斯琴格日乐，马端辉，包海鹰.玉米黑粉菌孢子粉的无机元素和氨基酸含量测定.菌物研究,2006,4(1):20-23.
[20]. Guus Bakkeren, J¨org K¨amper, Jan Schirawski. Sex in smut fungi: Structure, function andevolution of mating-type complexes. Fungal Genetics and Biology,2008,45: S15–S21.
[21]. Leo′n-Ram′rez CG, Cabrera-Ponce JL, Mart′nez-Espinoza AD, Herrera-Estrella L,Me′ndez L, Reynaga-Pen a CG, Ruiz-Herrera J. Infection of alternative host plant species byUstilago maydis. New Phytol,2004,164:337–346.
[22]. Banuett F. Pathogenic development in Ustilago maydis. In:Osiewacz HD (ed) Molecularbiology of fungal development. Marcel Dekker, New York, pp,2002:349–398.
[23]. Day, P.R., S.L. Anagnostakis, and J.E. Puhalla. Pathogenicity resulting from mutation at theb locus of Ustilago maydis. Proc. Natl. Acad. Sci,1971,68:533–535.
[24]. Bauer, R., Oberwinkler, F.&Vanky, K. Ultrastructural markers and systematics in smutfungi and allied taxa. Can. J. Bot,1997,75:273–1314.
[25]. Jo¨rg Ka¨mper, Regine Kahmann, Michael Bo¨lker. Insights from the genome of thebiotrophic fungal plant pathogen Ustilago maydis. NATURE,2006.
[26]. Basse, C.W., and Steinberg, G. Ustilago maydis, model system for analysis of the molecularbasis of fungal pathogenicity. Mol. Plant Pathol,2004,5:83–92.
[27]. Ka¨mper, J. et al. Insights from the genome of the biotrophic fungal plant pathogen Ustilagomaydis. Nature,2006,444:97–101.
[28]. Doehlemann, G., Wahl, R., Vranes, M., de Vries, R.P., K mper, J., and Kahmann, R.Establishment of compatibility in the Ustilago maydis/maize pathosystem. J Plant Physiol,2008,165:29–40.
[29].鄂文弟，王振华，张立国，张林，王霞，孙广权.玉米瘤黑粉病的研究进展.玉米科学,2006,14(1):153–157.
[30]. Regine Kahmann and J rg K mper. Ustilago maydis: how its biology relates to pathogenicdevelopment. New Phytologist,2004,164:31–42.
[31]. Caltrider PG, Gottlieb D. Effects of sugars on germination and metabolism of teliospores ofUstilago maydis. Phytopathology,1966,56:479–484.
[32]. Tripathi RK, Gottlieb D. Sequential biosynthesis of ribonucleic acids during germination ofteliospores of Ustilago maydis. Mycologia,1974,66:413–421.
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