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黄瓜花叶病毒(CMV)WL株系外壳蛋白编码基因的表达似乎能为烟草植株提供针对几种不同CMV株系感染的保护作用。

Expression of the gene encoding the coat protein of cucumber mosaic virus (CMV) strain WL appears to provide protection to tobacco plants against infection by several different CMV strains.

作者信息

Namba S, Ling K S, Gonsalves C, Gonsalves D, Slightom J L

机构信息

Department of Plant Pathology, Cornell University, New York State Agricultural Experiment Station, Geneva 14456.

出版信息

Gene. 1991 Nov 15;107(2):181-8. doi: 10.1016/0378-1119(91)90317-5.

Abstract

The gene (cp) encoding the coat protein (CP) of cucumber mosaic virus (CMV) strain WL (CMV-WL, which belongs to CMV subgroup II) was custom polymerase chain reaction (CPCR)-engineered for expression as described by Slightom [Gene 100 (1991) 251-255]. CPCR amplification was used to add 5'- and 3'-flanking NcoI sites to the CMV-WL cp gene, and cp was cloned into the expression vector, pUC18cpexp. This CMV-WL cp expression cassette was transferred into the genome of tobacco (Nicotiana tabacum cv. Havana 423) via the Agrobacterium T-DNA transfer mechanism. R0 plants that express the CMV-WL cp gene were subcloned, propagated, and challenge-inoculated with CMV-WL. Several R0 plant lines showed excellent protection against CMV-WL infection; however, plants found to accumulate the highest CP levels did not show the highest degree of protection. Thus in our case, CP levels appear not to be a useful predictor of the degree of protection. Plants from the best protected CMV-WL cp gene-expressing R0 tobacco lines were also inoculated with CMV strains belonging to the other major CMV subgroup (subgroup I), CMV-C and CMV-Chi, and compared in a parallel experiment with a transgenic tobacco plant line that expresses the CMV-C cp gene. Plants expressing the CMV-WL cp gene appeared to show a broader spectrum of protection against infection by the various CMV strains than plants expressing the CMV-C cp gene.

摘要

黄瓜花叶病毒(CMV)WL株系(CMV-WL,属于CMV亚组II)的外壳蛋白(CP)编码基因(cp)按照Slightom [Gene 100 (1991) 251-255]所述,通过定制聚合酶链反应(CPCR)进行工程改造以用于表达。利用CPCR扩增在CMV-WL cp基因的5'和3'侧翼添加NcoI位点,并将cp克隆到表达载体pUC18cpexp中。通过农杆菌T-DNA转移机制将这个CMV-WL cp表达盒转入烟草(Nicotiana tabacum cv. Havana 423)基因组中。对表达CMV-WL cp基因的R0植株进行亚克隆、繁殖,并接种CMV-WL进行挑战接种。几个R0株系对CMV-WL感染表现出优异的抗性;然而,发现积累最高CP水平的植株并未表现出最高程度的抗性。因此,在我们的研究中,CP水平似乎不是抗性程度的有效预测指标。来自最佳抗性的表达CMV-WL cp基因的R0烟草株系的植株也接种了属于其他主要CMV亚组(亚组I)的CMV株系,即CMV-C和CMV-Chi,并在平行实验中与表达CMV-C cp基因的转基因烟草株系进行比较。与表达CMV-C cp基因的植株相比,表达CMV-WL cp基因的植株似乎对各种CMV株系的感染表现出更广泛的抗性谱。

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