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Prothction of disease-resistant plants by cell engineering using host-specific toxins of pathogens

利用病原体宿主特异性毒素通过细胞工程保护植物的抗病性

基本信息

批准号：
10556007
负责人：
OTANI Hiroshi
金额：
$ 6.46万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10556007/
关键词：
host-specific toxin cell culture toxin-tolerant callus in vitro selection disease-resistant plant

项目摘要

Certain fungal pathogens are known to produce host-specific toxins (HSTs) that determine their host range and contribute to their virulence or pathogenicity. Recently, technology for breeding disease-resistant genotypes from susceptible plants by using HSTs is relatively prevalent. In many plants, somatic mutations generated through the growth of tissues or cells in culture release novel disease resistance characters, and HSTs are used to select in vitro for such characters.We found that Alternaria brassicicola, cucumber pathotype and tomato pathotype of Corynespora cassiicola, and Stemphylium vesicarium produce new HSTs named AB-, CCC-, CCT- and SV-toxins, respectively. AB-toxin was a protein and estimated to be 35 kDa, and CCT-toxin was a cyclic peptide with molecular weight of 2,780.CCT-toxin of C.cassiicola tomato pathotype was used for production of disease-resistant tomato. Tomato calli with green spots which have a high regeneration frequency were induced from hypocotyls and transferred to MS medium containing CCT-toxin. Although almost all calli became brown, some calli continued to survive even after incubation for 3 weeks. However, the toxin-tolerant calli lost ability to regenerate. On the other hand, A.alternata tomato pathotype produces a HST (AAL-toxin). Recently, the ESP1 which metabolizes AAL-toxin was cloned from black yeast Exophilia spinifera. Therefore, production of disease-resistant tomato by detoxification of AAL-toxin in ESP1-transgenic tomato was attempted. Transformation of ESP1 in tomato was performed by Agrobacterium tumefaciens containing binary vector with ESP1. However, transformants have not yet been obtained.

已知某些真菌病原体会产生宿主特异性毒素（HSTs），这些毒素决定了它们的宿主范围，并有助于它们的毒力或致病性。近年来，利用HSTs从易感植物中培育抗病基因型的技术比较普遍。在许多植物中，通过培养的组织或细胞生长产生的体细胞突变释放出新的抗病性状，HSTs被用于体外筛选这些性状。我们发现，甘蓝交替菌、黄瓜致病型、番茄致病型和水痘茎孢分别产生新的hst，命名为AB-、CCC-、CCT-和sv -毒素。ab -毒素是一种蛋白质，估计为35 kDa， cct -毒素是一种环肽，分子量为2780。利用卡氏弧菌番茄病原菌的cct毒素生产抗病番茄。用番茄下胚轴诱导出再生频率高的绿斑愈伤组织，并将其转移到含有cct毒素的MS培养基上。虽然几乎所有愈伤组织都变成褐色，但有些愈伤组织在孵育3周后仍能继续存活。然而，耐毒愈伤组织丧失了再生能力。另一方面，a.a alternata番茄病原菌产生HST （aal -毒素）。最近，从黑酵母中克隆出了代谢aal毒素的ESP1。因此，尝试利用转基因esp1番茄脱毒aal毒素生产抗病番茄。以农杆菌为载体，将ESP1在番茄中进行转化。然而，变形体尚未获得。