Studies on the utilization of useful genes isolated from the wild plant for control of virus diseases

克隆并测序了一种推测为无花果核糖体失活蛋白（CF-RIP）的cDNA。整个cDNA包含一个编码286个氨基酸的ORF，这些氨基酸共享核糖体失活蛋白（RIPS）的保守残基。CF-RIP与来自其他葫芦的4条rip共享54.8% -64.5%的同源性。RT-PCR分析表明，CF-RIP mRNA在黄瓜叶和茎组织中均有表达。从无花果树叶片中纯化了一种抗病毒蛋白，并对其进行了部分鉴定。柱层析、SDS-PAGE和周期性酸-希夫染色表明，figaren是一种糖蛋白，分子量为23 kDa。n端氨基酸测序结果表明，figaren含有s等位基因相关核糖核酸酶（RNases）的保守区。凝胶RNase实验表明无花果能消化酵母rna。荧光原位杂交结果显示，1μ g/ml的RNase和RNase（牛肉胰腺酶和RNase T1）对豇豆叶片中C…More MV感染有相似的抑制作用。Figaren和Rnases在5-500ng/ml对CMV局部病变的抑制作用相似。用ELISA法检测，接种前6、0小时经figen处理的烟草叶片中巨细胞病毒的积累均受到抑制。当烟草植株上部叶片经figaren处理后，10 min后接种巨细胞病毒，大部分烟草植株花叶病症状延迟5 ~ 7 d，部分植株无花叶病症状。接种了巨细胞病毒后再接种无花果的甜瓜原生质体中，巨细胞病毒rna和巨细胞病毒抗原的数量也随着时间的增加而增加。对照ELISA和局部病变试验表明，在接种巨细胞病毒前5 ~ 10 min，对烟草、甜瓜、豇豆和苋菜叶片上表面进行无花果处理的巨细胞病毒感染几乎完全被抑制。Figaren几乎完全抑制了未经处理的一半烟草叶片上的巨细胞病毒感染。无花果还能诱导烟草植株产生局部获得性抗性。少

A cDNA coding a putative ribosome-inactivating protein(CF-RIP) from Cucunis figarei was cloned and sequenced. The entire cDNA contains an ORF encoding a protein of 286 amino acids which share conserved residues for ribosome-inactivating proteins(RIPS). CF-RIP share 54.8-64.5% identity with four RIPs from other cucurbits. RT-PCR analysis indicated that CF-RIP mRNA was expressed in leaf and stem tissues of Cucumis figarei.An antiviral protein, designated figaren, was purified from leaves of C.figarei and partially characterized. Column chromatography, SDS-PAGE and periodic acid-Schiff staining revealed that figaren is a glycoprotein with a molecular weight of 23 kDa. N-terminal amino acid sequencing indicated that figaren contained the conserved region for the S-allele-associated ribonucleases(RNases). In-gel RNase assay showed that figaren digested yeast RNAs. Fluorescence in situ lybridization revealed that figaren and Rnases(beef pancrease and RNase T1)at 1μ g/ml similarly inhibited C … More MV infection in cowpea leaves. Figaren and Rnases at 5-500ng/mlhad similar inhibitory effect on local lesion formation by CMV.CMV accumulation detected by ELISA in tobacco leaves treated with figaren 6 or 0 hr before inoculation with CMV was suppressed. When upper leaves of tobacco plants were treated with figaren and inoculated 10 min later with CMV, mosaic symptoms were delayed for 5-7 days on most of tobacco plants, and some plants remained asymptomatic. The amount of CMV RNAs and CMV antigen in melon protoplasts inoculated with CMV and subsequently inoculated with figaren similarly increased with time as did that the control ELISA and local lesion assays indicated that CMV infection on the upper surfaces of the leaves of tobacco, melon, cowpea and Chenopodium amaranticolor whose lower surfaces had been treated with figaren 5-10 min before CMV inoculation was almost completely inhibited. Figaren almost completely inhibited CMV infection on the untreated halves of leaves of tobacco. Figaren also induced localized acquired resistance in tobacco plants. Less