Isolation of disease resistance related genes in mulberry plant by gene-tagging method

基因标记法分离桑树抗病相关基因

• 批准号: 08660066
• 负责人: NOZUE Masayuki
• 金额: $ 1.54万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660066/
• 关键词: Agrobacterium-mediated transformation; Bud blight; Disease resistance; Fusarium; Gene-tagging; Mulberry; Peroxidase; Polyphenol oxidase; 遺伝子タキング; 培養細胞

The bud blight of mulberry trees is one of the most destructive disease of mulberry stems. It is caused by Fusarium lateritium f.sp.mori, that produces sporodochia on the surface of dead mulberry stems. However, no such symptom appears in mulberry plant infected by non-pathogenic Fusarium to the host. Non-pathogenic fungi could not spread in the tissues of mulberry plant by disease resistance response of host cells. In order to investigate the mechanism of disease resistance in mulberry plant, this project is planned to characterize resistance-related gene that is isolated by DNA-tagging method. First of all, rapid and convenient method to estimate the ability to resist to Fusarium was established using mulberry cultured cells, that is necessary to select the transformed mutant cells. The efficient methods for Agrobacterium-mediated transformation in mulberry callus was investigated to obtain T-DNA tagged genes. Finally, peroxidase and polyphenol oxidase might be involved in tissue browning in the non-pathogenic Fusarium infected mulberry callus. The rapid and local browning in the infected tissue is the typical hypersensitive response in disease resistance. Both enzymes may play the important role in resistance response of mulberry plant against to fungi. It may be possible to isolate disease resistance-related genes of mulberry plant by the method established by the present study.

桑葚芽枯病是桑葚茎部最具破坏性的病害之一。它是由桑赤镰孢菌引起的，在桑葚茎表面产生孢母座。而感染非致病性镰刀菌的桑葚植株则无此症状。非病原真菌不能通过寄主细胞的抗病反应在桑葚组织中传播。为了研究桑葚抗病机理，本项目拟对利用DNA标签法分离的抗病相关基因进行鉴定。首先，建立了桑葚培养细胞抗镰刀菌能力的快速、简便的鉴定方法，为筛选转化突变体细胞奠定了基础。为获得T-DNA标记的基因，研究了农杆菌介导的桑葚愈伤组织转化方法。过氧化物酶和多酚氧化酶可能参与了非致病性镰刀菌感染的桑葚愈伤组织的布朗宁。侵染组织迅速而局部的布朗宁是典型的抗病过敏反应。这两种酶可能在桑葚对真菌的抗性反应中起重要作用。本研究建立的方法为分离桑葚抗病相关基因提供了可能。

项目成果

野末雅之: "クワ培養細胞を用いたクワ芽枯病菌の病原性評価法の確立" 信州大学繊維学部農場研究報告. 16. 5-9 (1997)

Masayuki Nozue：“建立利用培养的桑树细胞评价桑树芽枯病致病性的方法”信州大学纺织学部农场研究报告16. 5-9 (1997)。

野末 雅之 他5名: "クワ培養細胞を用いたクワ芽枯病菌の病原性評価法の確立" 信州大学繊維学部農場研究報告. 16. 5-9 (1997)

Masayuki Nozue等5人：“利用培养的桑树细胞评价桑树芽枯病致病性的方法的建立”信州大学纺织学部农场研究报告16. 5-9 (1997)。

Nozue, M., Cai, W., Ishii, C., Shioiri, H., Kojima, M.and Saito, H.: "In vitro Assay for pathogenicity in Fusarium using mulberry cultured cells" Bull.Exp.Farm Coll.Agr., Shinshu Univ.16. 5-9 (1997)

Nozue, M.、Cai, W.、Ishii, C.、Shioiri, H.、Kojima, M. 和 Saito, H.：“使用桑培养细胞对镰刀菌的致病性进行体外测定”Bull.Exp.Farm Coll。