立枯丝核菌胞壁降解酶及其在致病中的作用-作物遗传育种专业论文.docxVIP

张红；立桔丝核葡匏壁降解酶及其在致病中的作用立枯丝核菌胞壁降解酶及其在致病中的作用 张红；立桔丝核葡匏壁降解酶及其在致病中的作用 立枯丝核菌胞壁降解酶及其在致病中的作用 摘要 采用改良的Marcus培养液培养立枯丝核菌(Rhizoctonia solani Kinm)，滤液 经离心，留上清液即粗酶液。租酶液具有cx、PG、PMG、PG-TE和PMTE等5种 胞壁降解酶的活性。其中前三种酶活性较高，后两种酶活性较低。 立枯丝核菌产生胞壁降解酶的最佳条件是：培养时间为6d；培养温度为28。C： 培养液初始姑I为5左右：培养时不需振荡。 以病菌接种成株期水稻倒二叶鞘，发病后分别检测桔自、褐色和褪绿部位组 织中胞壁降解酶的活性。三部位组织中均有上述5种胞壁降解酶存在，其中PG 活性最高，达21．123 U／tag：其次是PMG；而PGTE和PMTE活性很低．约为O．1 U／rag。从不同部位缀织来看，褪绿部胞壁降解酶活性最高，达13．417 U／mg．其次 是褐色部，酶活性为l 1．765 U／mg，再次为枯白部。这表明病菌胞壁降解酶在病斑 形成和扩展中起重要作用。 立桔丝核菌胞壁降解酶对水稻组织细胞具有显著鹩破坏作用。粗酶液处理能 引起水稻苗期叶鞘黄化，呈水溃状。显微观察表明，粗酶液可破坏愈伤组织细胞， 酶液处理24h后细胞壁扭曲变形、壁部分消解或出现微纤丝；处理48h后细胞壁 破裂，细胞质大量外淹。通过电镜观察发现。粗酶液对菌期水稻叶鞘组织细胞也 有破坏，酶液处理36h后细胞出现质壁分离、细胞壁部分断裂、细胞器分辨不清、 叶绿体和线粒体受损等现象。 立桔丝核菌胞壁降解酶对水稻时鞘细胞膜也有明显损伤作用。酶液处理后细 胞透性改变，其损伤影响混合酶大于单一酶，果胶酶大于纤维素酶。 立枯丝核菌不同菌株产生胞壁降解酶的能力有一定差异。通过比较发现，菌 株产生胞壁降解酶的能力与其对水稻植抹的致病力闻存在极显著正相关。这揭示 了立枯丝核菌胞壁降解酶在病菌致病过程中发挥着重要作用，是该病重要的致病 因子。 关键词：立桔丝核菌，胞壁降解酶，产生条件，致病作用，水稻 扬州大学硕士学位论文Cell-wall 扬州大学硕士学位论文 Cell-wall degrading enzymes produced by Rhizoctonia solani and their pathogenicity to rice plants Abstract nle crude cell—wall degrading enzymes(CWDEsl produced by Rhizoctonia solani Knhn in improved Marcus’S medium were extracted when the cultured filtrate WaS centrifugalled and then the superuatant was reserved．Polygalacturonase(PO)，eellulase (Cx)，pectin methylgalacturonase(PMG)，polygalacturonic acid trans-eliminase(PGTE) and pectin methyl trails—eliminase fPMTE)were determined in the crude enzynles． Among the CWDEs，the activities of PG，Cx and PMG were significantly higher than the others’． nle culture condition for the production ofRhizoctonia solani CWDEs was studied． It Was appropriate for the CWDEs production as follows：ne strains was cultured for 6 days aI 28℃in the improved Marcns’s medium，in which pH value Was 5，without shaking． Rice plants were inoculated by pMcing a block of the mycelia on the sheath．The activities of CWDEs were determined respectively in three parts of lesion，such as the withered，the brown and the fad吨．There were five CWDEs mentioned ab