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使用单克隆抗体测定感染番茄叶片中番茄褐腐病菌的生物量。

Use of monoclonal antibodies to determine biomass of Cladosporium fulvum in infected tomato leaves.

作者信息

Karpovich-Tate N, Spanu P, Dewey F M

出版信息

Mol Plant Microbe Interact. 1998 Jul;11(7):710-6. doi: 10.1094/MPMI.1998.11.7.710.

Abstract

A monoclonal antibody, OX-CH1, was raised against surface washings of Cladosporium herbarum. This antibody recognizes an epitope that is found in various fungi belonging to the genus Cladosporium, including C. fulvum, the causal agent of tomato leaf mold. The epitope is present at comparable levels in two different races of C. fulvum and in transgenic isolates derived from them. The epitope is heat-and protease-resistant but sensitive to oxidation with periodate and it is constitutively expressed in C. fulvum grown in pure culture and on the plant. C. fulvum can be detected in infected tissues at levels starting from around 1 mg fresh weight of fungus per g fresh weight of leaf tissue. Noninfected tomato leaves do not cross-react with OX-CH1. We have developed an enzyme-linked immunosorbent assay (ELISA) for fungal biomass in tomato leaves and compared it with the assay based on measurements of beta-glucuronidase (GUS) activity in tissues infected with a transgenic isolate of C. fulvum race 4 carrying a uidA gene; the two assays give similar results.

摘要

制备了一种针对草本枝孢菌表面洗涤物的单克隆抗体OX-CH1。该抗体识别一种表位,这种表位存在于枝孢菌属的多种真菌中,包括引起番茄叶霉病的番茄黄枝孢菌。在番茄黄枝孢菌的两个不同小种及其衍生的转基因分离物中,该表位的含量相当。该表位对热和蛋白酶具有抗性,但对高碘酸盐氧化敏感,并且在纯培养物和植物上生长的番茄黄枝孢菌中组成性表达。在感染组织中可以检测到番茄黄枝孢菌,其含量约为每克新鲜叶片组织1毫克新鲜真菌重量。未感染的番茄叶片与OX-CH1不发生交叉反应。我们开发了一种用于检测番茄叶片中真菌生物量的酶联免疫吸附测定法(ELISA),并将其与基于对携带uidA基因的番茄黄枝孢菌4号小种转基因分离物感染组织中β-葡萄糖醛酸酶(GUS)活性测量的测定法进行了比较;两种测定法给出了相似的结果。

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