Immunological detection of endo polygalacturonase secretion by Fusarium oxysporum in plant tissue and sequencing of its encoding gene
Immunological detection of endo polygalacturonase secretion by Fusarium oxysporum in plant tissue and sequencing of its encoding gene
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植物组织中尖镰孢内切多聚半乳糖醛酸酶分泌的免疫学检测及其编码基因测序
DOI：
10.3186/jjphytopath.64.7
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
I. Yamaguchi
中科院分区：
文献类型：
--
作者：
T. Arie;S. Gouthu;S. Shimazaki;T. Kamakura;M. Kimura;M. Inoue;K. Takio;A. Ozaki;K. Yoneyama;I. Yamaguchi
尖孢镰刀菌（Fusarium oxysporum）是一种土传植物病原真菌，其产生的内切多聚半乳糖醛酸酶（endo polygalacturonase，PG）被认为与病害的发生有关。从F. oxysporum F.番茄小种2.用该抗体进行直接组织印迹免疫结合试验（DT-IBA），证实了病原菌在寄主植物茎中产生PG。根据该菌PG蛋白的部分氨基酸残基信息设计简并引物，通过PCR扩增获得部分PG编码基因。利用TAIL-PCR技术，克隆了PG基因的全序列。该基因全长1318 bp，编码371个氨基酸，其中含有22个氨基酸的信号肽，开放阅读框被47、51、50和54 bp的内含子打断。推导的成熟蛋白氨基酸序列与F.串珠状菌PG、炭旋孢霉PGNI、尼日尔曲霉PG、黑曲霉A. PGI和核盘菌PGI。
Fusarium oxysporum, a soilborne phytopathogenic fungus, produces endo polygalacturonase (PG) which has been suggested to be related to disease development. A polyclonal antibody (PAb) APG1 was prepared against purified PG from F. oxysporum f. sp. lycopersici race 2. By direct tissue-blotted immunobinding assay (DT-IBA) with the antibody, PG production by the pathogen in the stem of the host plant was confirmed. Degenerate primers were designed based on partial amino acid residue information for the PG protein from the fungus: part of the PG-encoding gene was obtained by PCR. Using TAIL-PCR, the complete gene encoding PG was cloned and sequenced. The structural gene comprises 1318 bp coding for 371 amino acids with a putative signal peptide of 22 amino acids, and the open reading frame is interrupted by four introns of 47, 51, 50, and 54bp. The deduced amino acid sequence of the mature protein showed 82.9, 29.0, 27.6, 14.2, and 26.9% homology with those of F. moniliforme PG, Cochliobolus carbonum PGNI, Aspergillus niger PG, A. oryzae PG, and Sclerotinia sclerotiorum PGI, respectively.