Agrawal G K, Rakwal R, Jwa N S
Research Laboratory for Agricultural Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal.
Biochem Biophys Res Commun. 2000 Nov 19;278(2):290-8. doi: 10.1006/bbrc.2000.3781.
Strategies evolved by plants to counteract a variety of biotic/abiotic stresses include induction of genes encoding pathogenesis-related (PR) protein, in particular the PR class 1 (PR1) gene family, widely used in stress response studies. In spite of its immense importance as a PR family member, and an accepted gene marker in plant disease/defense in dicots, little is known about rice PR1 genes. Recently, we cloned and characterized the first OsPR1a (rice acidic PR1) gene (Agrawal et al. (2000) Biochem. Biophys. Res. Commun. 274, 157-165). Here, we report characterization of a rice basic PR1 (OsPR1b) gene, identified from screening a cDNA library prepared from jasmonic acid (JA)-treated rice seedling leaf, providing detailed and valuable insights into rice PR1 gene expression. The deduced amino acid sequence of OsPR1b reveals only 63.1% homology with the OsPR1a protein, whereas Southern blot analyses indicate that OsPR1b is a multigene family. The JA-inducible OsPR1b gene was also up-regulated by salicylic acid (SA), abscisic acid (ABA), and kinetin (KN). Furthermore, protein phosphatase inhibitors, cantharidin (CN) and endothall (EN) strongly induced the OsPR1b transcript. However, OsPR1b was not cut-responsive, diagrammatically opposite to cut inducibility of OsPR1a. This induction was light-, time-, and dose-dependent, as demonstrated by using, JA, CN, and EN, and completely inhibited by cycloheximide, but not by tetracycline. The simultaneous application of SA, and ABA, with JA, respectively, showed almost complete inhibition of the JA-induced OsPR1b transcript by 200 microM SA or ABA, but not by 100 microM concentrated solutions, suggesting a potential interaction among JA, SA, and ABA, whereas KN dramatically enhanced JA-induced OsPR1b transcript upon simultaneous application. Moreover, a simultaneous application of staurosporine enhances JA-, CN-, and EN-induced OsPR1b transcript, in particular with CN. Finally, a comparative analysis with the OsPR1a gene gives us insight into the differential regulation of the PR1 gene family, while proposing OsPR1 genes as important gene markers in rice, with potential use(s) in analyzing plant defense responses.
植物为应对各种生物/非生物胁迫而进化出的策略包括诱导编码病程相关(PR)蛋白的基因,特别是PR1类(PR1)基因家族,该家族在胁迫反应研究中被广泛应用。尽管作为PR家族成员及其在双子叶植物的植物病害/防御中作为公认的基因标记具有极其重要的意义,但关于水稻PR1基因却知之甚少。最近,我们克隆并鉴定了首个OsPR1a(水稻酸性PR1)基因(Agrawal等人,(2000年)《生物化学与生物物理研究通讯》274,157 - 165)。在此,我们报告了从茉莉酸(JA)处理的水稻幼苗叶片制备的cDNA文库筛选中鉴定出的水稻碱性PR1(OsPR1b)基因的特征,为水稻PR1基因表达提供了详细且有价值的见解。OsPR1b推导的氨基酸序列与OsPR1a蛋白仅显示63.1%的同源性,而Southern印迹分析表明OsPR1b是一个多基因家族。JA诱导的OsPR1b基因也被水杨酸(SA)、脱落酸(ABA)和激动素(KN)上调。此外,蛋白磷酸酶抑制剂,斑蝥素(CN)和克芜踪(EN)强烈诱导OsPR1b转录本。然而,OsPR1b对切割无反应,这与OsPR1a的切割诱导性形成鲜明对比。如使用JA、CN和EN所证明的,这种诱导是光依赖性、时间依赖性和剂量依赖性的,并且被放线菌酮完全抑制,但不被四环素抑制。分别将SA和ABA与JA同时应用,显示200 microM的SA或ABA几乎完全抑制JA诱导的OsPR1b转录本,但100 microM浓缩溶液则无此作用,这表明JA、SA和ABA之间可能存在相互作用,而KN在同时应用时显著增强JA诱导的OsPR1b转录本。此外,同时应用星形孢菌素可增强JA、CN和EN诱导的OsPR1b转录本,特别是与CN同时应用时。最后,与OsPR1a基因的比较分析使我们深入了解PR1基因家族的差异调控,同时提出OsPR1基因作为水稻中的重要基因标记,在分析植物防御反应方面具有潜在用途。