水稻白叶枯病菌的血清学研究

SEROLOGICAL STUDIES ON THE PATHOGEN OF BACTERIAL BLIGHT OF RICE(XANTHOMONAS CAMPESTRIA PV路ORYZAE DYE)

用戊二醛固定的5个水稻白叶枯菌株制备了5个抗血清,利用琼脂双扩散、试管凝集、免疫电泳和酶联免疫吸附试验研究了107个水稻白叶枯菌株血清学反应的差异,将它们分为三个血清型。属于Ⅰ型的菌株(OS-225、F_4等)占供试菌株的95%,分布于全国各稻区;归属于Ⅱ型的菌株(OS 209、OS-109等)占5.6%,主要来自南方边远地区;Ⅱ型仅1个菌株(G_8),占0.9%,来自广西东部;其中还有两个菌株和上述抗血清均不能反应,暂不能归型,来自福建。血清型和致病型之间的相关性不用戊二醛固定、热处理等5种不同方法处理菌体后制备的抗血清之间,表现出一致的反应特异性和相似的“型”专化性。免疫电泳结果表明,不同血清型的免疫源组成是不同的,Ⅰ型仅具中性免疫源,Ⅱ型具中性及偏碱性免疫源,而Ⅱ型则具中性及偏酸性两种免疫源。比较了反向间接血凝、酶联免疫吸附和免疫荧光反应等方法检测病菌的灵敏度,以免疫荧光试验(IF)为最灵敏,可检测到每毫升10~3细胞,ELISA其次(10^(4-3)cell/ml),反向间接血凝法为10^(6-7)cells/ml,双扩散法最差,只有在高达10~8 cells/ml时才有阳性反应。

Five antisera were produced against 5 isolates of Xanthomonas campestrts pv. oryzae after fixed by glutaradehyde. Three serotypes designated serotypes Ⅰ, Ⅱ and Ⅲ were distinguished. By ■eans of Ouchterlony double diffusion test (ODD), tube agglutination test, immunoelectrophoresis (IE) and enzyme lin■ed immunosorbent assay (ELISA) with above antisera, among 105 isolates, 93% belonged to serotype I, distributed all over China; 6% to serotype Ⅱ, mainly in the southern rice area; only one isolate to serotype Ⅲ which came from Guangxi autononous region.Five immunogens produced from isolate (OS-225) by different treatments, heated all cells, glutaraldehyde-fixed, membrane protein complex, glycoprotein and crude glycoprotein were compared. It showed that all of these had a similar specificity. In ODD test, antiserum to each of five immunogens only reacted with the isolates of serotype I, but did not react with isolates of other two serotypes.None of above mentioned antisera reacted with other species of Xanthomonas or the bacteria of other genera.The results of IE showed that those isolates of serotype I only had neutral immunogen, serotype Ⅱ had both neutral and alkaline immunogens and serotype Ⅲ had both neutral and acidic immunogens.Four serological methods were compared for detection of samples infected by X.c.pv. oryzae. The highest sensitivity was found in immunofluorescence (IF) test (10~3 cells/ml), and followed by ELISA and indirect Hemagglutination test. The IF and ELISA could be applied effectively for detection of the inoculum in different plant materials and study of the ecology of bacteria.