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Participation of the Cowpea mosaic virus protease in eliciting extreme resistance.

机译:pea豆花叶病毒蛋白酶参与引起极端抗性。

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Extreme resistance of Arlington line cowpea (Vigna unguiculata) to Cowpea mosaic virus (CPMV) is under control of a dominant locus designated Cpa. We transiently expressed, using Tomato bushy stunt virus (TBSV) vectors and Agrobacterium tumefaciens, in nearly isogenic Cpa/Cpa and cpa/cpa cowpea lines, sequences from RNA1, the larger of two CPMV genomic RNAs. Activation of a Cpa-specific response mapped to the CPMV 24K protease (24KPro). Mutational analysis of the 24KPro gene implicated protease activity, rather than 24KPro structure, in Cpa-mediated recognition of CPMV invasion. A 24KPro with alanine replacing the active site cysteine [24KPro(C-A)], but not wildtype 24KPro, accumulated after agroinfiltration of the corresponding binary vector constructions into Cpa/Cpa cowpea. In cpa/cpa cowpea, both protease versions accumulated, with 24KPro(C-A) in greater abundance. Thus, enzymically active 24KPro was recognized by both cowpea genotypes, but in Cpa/Cpa cowpea the suppression of 24KPro accumulation was very strong, consistent with extreme resistance to CPMV.
机译:阿灵顿品系cow豆(Vigna unguiculata)对pea豆花叶病毒(CPMV)的极端抗性在称为Cpa的优势基因座的控制下。我们使用番茄浓密的特技病毒(TBSV)载体和根癌农杆菌在几乎同基因的Cpa / Cpa和cpa / cpa cow豆系中瞬时表达了来自RNAi的序列,这是两个CPMV基因组RNA中较大的一个。映射到CPMV 24K蛋白酶(24KPro)的Cpa特异性应答的激活。在Cpa介导的CPMV入侵识别中,对24KPro基因的突变分析涉及蛋白酶活性,而不是24KPro结构。在将相应的二元载体构建体浸入Cpa / Cpa pea豆后,用丙氨酸替代活性位点半胱氨酸[24KPro(C-A)]而不是野生型24KPro积累了24KPro。在cpa / cpa cow豆中,两种蛋白酶都积累在一起,其中24KPro(C-A)的含量更高。因此,两种cow豆基因型均识别出具有酶活性的24KPro,但在Cpa / Cpa cow豆中,对24KPro积累的抑制作用非常强,与对CPMV的极强抗性相一致。

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