Studies on the production of Porphyrae yezoensis reeds tolerant to the red rot disease by the gene transduction of anti-Pythium protein

抗腐霉蛋白基因转导生产抗红腐病紫菜芦苇的研究

• 批准号: 13660186
• 负责人: KAMEI Yuto
• 金额: $ 2.18万
• 依托单位: Saga University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660186/
• 关键词: the Ariake Sea; Porphyrae yezoensis; red rot disease; Pythium porphyrae; disease-tolerant Porphyrae yezoensis; antifungal protein; cell wall degrading activity; gene transduction; 真菌ピシウム; ノリプロトプラスト; ノリ細胞壁分解酵素; 組換え体粗酵素; 組換え作物; 抗ピシウムタンパク; 作用機序

In the present study, we conducted to screen the antifungal proteins against Pythium porphyare to establish the new breeds of Pophyrae yezoensis tolerant to the red rot disease, and isolated 13 strains of bacteria with the cell wall degrading activity to P. porphyare from the Ariake Sea. Pseudomoas sp. PE2, the most potent active bacterium showed chitin, β-1,3;1,4-glucan, and β-1,3-glucan degrading activities and then these genes were cloned by the expression of these enzymes. When we analyzed these genes, these proteins have been proved to have the functional regions with a variety of binding activities to several polysaccharides and specifically bound to some polysaccharides. Among these enzymes, β-1,3(4)-glucanase A has been found to be most important for the cell wall degradation. More over, when we purified and characterized the anti-P. porphyare protein (SAP) from the culture supernatant of the marine bacterial isolate, Sreptomyces sp. AP77, SAP was proved to have three subunits and the homologies of these subunits are very high to the known proteins. The mode of action of SAP seems to be related to the permeability of cell membrane of P.porhyrae.Thus, the present study proves that above enzymes and antifungal proteins are very useful and expectable to establish the disease-tolerant breeds of P.yezoensis by the gene transduction of those proteins.

为建立抗红腐病的条斑紫菜新品种，本研究从有明海域分离到13株对紫菜腐霉具有细胞壁降解活性的细菌，并筛选出抗紫菜腐霉的蛋白质。拟单胞菌PE 2是一株具有几丁质、β-1，3-葡聚糖和β-1，3-葡聚糖降解活性的细菌，通过表达这些酶，克隆了这些基因。通过对这些基因的分析，发现这些蛋白质具有与多种多糖结合的功能区域，并与某些多糖特异性结合。在这些酶中，已经发现β-1，3（4）-葡聚糖酶A对于细胞壁降解是最重要的。此外，我们从海洋细菌分离物Sreptomatrium sp.AP77的培养上清中纯化并鉴定了抗卟啉单胞菌蛋白（SAP），结果表明SAP含有3个亚基，且与已知蛋白的同源性很高。SAP的作用方式可能与条斑紫菜细胞膜的通透性有关，因此，本研究证明上述酶和抗真菌蛋白对条斑紫菜抗病育种具有重要的应用价值。

项目成果

Molecular cloning, sequencing and expression of the gene encoding a novel chitinase A from a marine bacterium, Pseudomonas sp PE2, and its domain structure

• DOI: 10.1007/s00253-002-1154-x
• 发表时间: 2003-04-01
• 期刊: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
• 影响因子: 5
• 作者: Kitamura, E;Kamei, Y
• 通讯作者: Kamei, Y

protein from the marine bacterium, Streptomyces sp. AP77 is specific for Pythium porphyrae, a causative agent of red rot disease in Porphyra spp.

来自海洋细菌链霉菌的蛋白质。

• 发表时间: 2002
• 期刊: Applied and Environmental Microbiology 68
• 作者: J.H.Woo;E.Kitamura;E.Myouga;Y.Kamei
• 通讯作者: Y.Kamei

Etsushi Kitamura, Yuto Kamei: "Molecular cloning, sequencing and expression of the gene encoding a novel chitinase A from a marine bacterium, Pseudomonas sp. SE2 and its domain structure"Applied Microbiology & Biotechnology. (in press). (2003)

Etsushi Kitamura、Yuto Kamei：“来自海洋细菌假单胞菌 SE2 的新型几丁质酶 A 编码基因的分子克隆、测序和表达及其结构域”应用微生物学

Polysaccharolytic activities of bacterial enzymes that degrade the cell walls of Pythium porphyrae, a causative fungus of red rot disease in Porphyra yezoensis

降解紫菜腐霉细胞壁的细菌酶的多糖分解活性，紫菜腐霉是条斑紫菜红腐病的致病真菌

• 发表时间: 2002
• 期刊: Fisheries Sciences 68
• 作者: E.Kitamura;H.Myouga;Y.Kamei
• 通讯作者: Y.Kamei

Polysaccharolytic activities of bacterial enzymes that degrade the cell walls of Pythium porphyrae, a causative fungus of red rot disease in Porphyra yezoensis.

降解紫菜腐霉细胞壁的细菌酶的多糖分解活性，紫菜腐霉是条斑紫菜红腐病的致病真菌。

• 发表时间: 2002
• 期刊: Fisheries Sciences 68
• 作者: E.Kitamura;H.Myouga;Y.Kamei
• 通讯作者: Y.Kamei