Bradshaw H D, Hollick J B, Parsons T J, Clarke H R, Gordon M P
Department of Biochemistry, University of Washington, Seattle 98195.
Plant Mol Biol. 1990 Jan;14(1):51-9. doi: 10.1007/BF00015654.
When the lower leaves of hybrid poplar trees are mechanically wounded, several novel mRNAs accumulate in the unwounded upper leaves (Parsons TJ, Bradshaw HD, Gordon MP: Systemic accumulation of specific mRNAs in response to wounding in poplar trees, Proc Natl Acad Sci USA, in press). A partial cDNA clone corresponding to a transcript from the wound-responsive gene designated win 3 (wound-inducible) has been cloned by differential hybridization to 32P-labelled cDNA from the leaves of wounded trees. Northern blots show a large accumulation of win 3 transcripts in the unwounded leaves of wounded trees. Southern blot analysis of poplar DNA suggests that win 3 is a member of a multigene family. The nucleotide sequences of several win 3 cDNA clones have been determined, indicating that at least three win 3 gene family members are transcribed. A genomic clone of a win 3 gene family member has been isolated and a 1.5 kb Hind III fragment containing the predicted protein-coding and 5' upstream regions has been sequenced. The putative win 3 gene product is similar to the major soluble proteins of sweet potato tubers, sporamin A and sporamin B. Both Win3 and the sporamins share significant amino acid sequence identity with Kunitz-type trypsin inhibitors from legume seeds. The Kunitz family of proteinase inhibitors thus joints three other proteinase inhibitor families which are systemically responsive to wounding.
当杂交杨树的下部叶片受到机械损伤时,几种新的信使核糖核酸(mRNA)会在未受伤的上部叶片中积累(帕森斯TJ、布拉德肖HD、戈登MP:杨树受伤后特定mRNA的系统性积累,《美国国家科学院院刊》,即将发表)。通过与来自受伤树木叶片的用32P标记的互补脱氧核糖核酸(cDNA)进行差异杂交,克隆了一个与伤口反应基因win 3(伤口诱导型)转录本相对应的部分cDNA克隆。Northern印迹显示,win 3转录本在受伤树木未受伤叶片中大量积累。杨树DNA的Southern印迹分析表明,win 3是一个多基因家族的成员。已确定了几个win 3 cDNA克隆的核苷酸序列,表明至少有三个win 3基因家族成员被转录。分离出了一个win 3基因家族成员的基因组克隆,并对一个包含预测的蛋白质编码区和5'上游区域的1.5 kb Hind III片段进行了测序。推测的win 3基因产物与甘薯块茎的主要可溶性蛋白质、sporamin A和sporamin B相似。Win3和sporamins与豆科种子的Kunitz型胰蛋白酶抑制剂都有显著的氨基酸序列同一性。因此,Kunitz蛋白酶抑制剂家族与其他三个对伤口有系统性反应的蛋白酶抑制剂家族联系在一起。