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中国水稻条斑病细菌avrBs3/PthA家族基因的克隆和序列分析 被引量:19

Cloning and Sequence Analysis of Diverse Members of avrBs3/PthA Family of Xanthomonas oryzae pv.oryzicola
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摘要 水稻条斑病细菌(Xanthomonasoryzaepv.oryzicola,Xooc)和水稻白叶枯病菌(X.oryzaepv.oryzae,Xoo)是稻黄单胞菌种下的致病变种,是水稻上的模式病原菌。用Xoo无毒基因avrXa3对应的NLS和AD区域的DNA片段构建探针,筛选Xooc基因文库,再通过酶切归类和Southern杂交分析,从Xooc中获得2个avBs3r/PthA家族基因克隆,分别被命名为avr/Pth13和avr/Pth14。序列测定结果显示,它们与avrBs3PthA家族成员,几乎一样的5′端和3′端、1个亮氨酸拉链(leucinezipper,LZ)、3个核定位信号(nuclearlocalizationsignal,NLS)和1个酸性转录活化域(acidictranscriptionalactivationdomain,AD),不同的只是102bp重复单元的重复次数不等:avr/Pth13基因102bp重复5.5次,avr/Pth14基因为19.5次。蛋白质水平上同列比较发现,AvrBs3/PthA家族成员34aa重复单元的第12和第13位氨基酸为HD的至少需要3个,才可能满足毒性或无毒性功能。将avr/Pth13和avr/Pth14基因分别导入Xoo菌株PXO99A中之后,在抗水稻白叶枯病近等基因系上测定其致病性,发现avr/Pth13基因增强了PXO99A的致病能力,而avr/Pth14基因则减弱了PXO99A的致病力。以上结果提示,Xooc与Xoo一样,都存在avrBs3/PthA基因家族的无毒基因。从Xooc中克隆avr/Pth13和avr/Pth14基因在国内外尚属首次,? Xanthomonas oryzae pv.oryzicola (Xooc) and X.o.pv.oryzae (Xoo) are two pathovars on rice. An EcoRI-BamHI DNA fragment, corresponding to the coding region NLS and AD of AvrXa3 of Xoo, was used as a probe to screen the genomic library of Xooc. Twenty three avrBs3/PthA gene clones were obtained after positive clones of the library were digested by EcoRI endoenzyme and proved by Southern blot with the probe. Sequencing randomly selected clones, avr/Pth13 and avr/pth14, indicated that they were the members of avrBs3/PthA family of Xanthomonas pathogenic bacteria. The two putative proteins were similar as other AvrBs3/PthA, possessing the same 5′- and 3′- terminals, one leucine zipper (LZ), three nuclear localization signals (NLSs) and one acidic transcriptional activation domain (AD). However, the difference of the gene with other avrBs3/PthA family members was the repeat numbers of 34-amino-acid unit coded by 102 bp repeat. The repeat numbers of 102 bp unit was 5.5 in avr/Pth13 gene and 19.5 in avr/Pth14。Alignments of 12-13th residues in 34-amino-acid unit of Avr/Pth13 and Avr/Pth14 with those of other reported AvrBs3/PthA proteins demonstrated hypothetically that at least three HD residues in 12-13th position of 34-amino-acid unit were necessary for avirulence or/and virulence of the proteins. Pathogenicity of strain PXO99A containing avr/Pth13 and avr/Pth14 gene, respectively, on near isogenic lines (NILs) and rice cultivars with Xoo R genes showed that the virulence of PXO99A was strengthened by avr/Pth13 gene, and reduced by avr/Pth14. All the results confirmed that diverse avrBs3/PthA genes existed in Xooc as that in Xoo determining virulence in bacterial streak and blight diseases of rice. It is the first report that the avrBs3/PthA genes were cloned from Xooc and avr/Pth13 is the smallest gene in the avrBs3/PthA family up to now。
出处 《中国农业科学》 CAS CSCD 北大核心 2005年第5期929-935,共7页 Scientia Agricultura Sinica
基金 国家"863"计划(2004AA214093) 国家自然科学基金(30370926) 江苏省科技攻关项目(BE2002303)资助
关键词 中国 水稻条斑病细菌 avrBs3/PthA基因家族 基因克隆 序列分析 致病性 Xanthomonas oryzae pv. oryzicola avrBs3/PthA family Southern blot Pathogenicity
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参考文献16

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