Molecular Crop Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Omics Sciences and Bioinformatics Center, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
BMC Plant Biol. 2019 Nov 6;19(1):472. doi: 10.1186/s12870-019-2086-2.
Isocitrate lyase (ICL) is a key enzyme in the glyoxylate cycle. In a previous study in rice, the expression of the ICL-encoding gene (OsICL) was highly induced by salt stress and its expression was enhanced in transgenic rice lines overexpressing OsCam1-1, a calmodulin (CaM)-encoding gene. CaM has been implicated in salt tolerance mechanisms in plants; however, the cellular mechanisms mediated by CaM are not clearly understood. In this study, the role of OsICL in plant salt tolerance mechanisms and the possible involvement of CaM were investigated using transgenic plants expressing OsICL or OsCam1-1.
OsICL was highly expressed in senesced leaf and significantly induced by salt stress in three OsCam1-1 overexpressing transgenic rice lines as well as in wild type (WT). In WT young leaf, although OsICL expression was not affected by salt stress, all three transgenic lines exhibited highly induced expression levels. In Arabidopsis, salt stress had negative effects on germination and seedling growth of the AtICL knockout mutant (Aticl mutant). To examine the roles of OsICL we generated the following transgenic Arabidopsis lines: the Aticl mutant expressing OsICL driven by the native AtICL promoter, the Aticl mutant overexpressing OsICL driven by the 35SCaMV promoter, and WT overexpressing OsICL driven by the 35SCaMV promoter. Under salt stress, the germination rate and seedling fresh and dry weights of the OsICL-expressing lines were higher than those of the Aticl mutant, and the two lines with the icl mutant background were similar to the WT. The F/F and temperature of rosette leaves in the OsICL-expressing lines were less affected by salt stress than they were in the Aticl mutant. Finally, glucose and fructose contents of the Aticl mutant under salt stress were highest, whereas those of OsICL-expressing lines were similar to or lower than those of the WT.
OsICL, a salt-responsive gene, was characterized in the transgenic Arabidopsis lines, revealing that OsICL expression could revert the salt sensitivity phenotypes of the Aticl knockout mutant. This work provides novel evidence that supports the role of ICL in plant salt tolerance through the glyoxylate cycle and the possible involvement of OsCam1-1 in regulating its transcription.
异柠檬酸裂解酶(ICL)是乙醛酸循环中的关键酶。在之前对水稻的研究中,ICL 编码基因(OsICL)的表达受盐胁迫高度诱导,并且在过表达钙调素(CaM)编码基因 OsCam1-1 的转基因水稻品系中表达增强。CaM 已被牵连到植物的耐盐机制中;然而,CaM 介导的细胞机制尚不清楚。在这项研究中,使用表达 OsICL 或 OsCam1-1 的转基因植物,研究了 OsICL 在植物耐盐机制中的作用以及 CaM 的可能参与。
OsICL 在衰老叶片中高度表达,在三个 OsCam1-1 过表达的转基因水稻品系以及野生型(WT)中均被盐胁迫显著诱导。在 WT 幼叶中,尽管 OsICL 表达不受盐胁迫影响,但所有三个转基因系均表现出高度诱导的表达水平。在拟南芥中,盐胁迫对 AtICL 敲除突变体(Aticl 突变体)的萌发和幼苗生长有负面影响。为了研究 OsICL 的作用,我们生成了以下转基因拟南芥系:由天然 AtICL 启动子驱动表达 OsICL 的 Aticl 突变体、由 35SCaMV 启动子驱动表达 OsICL 的 Aticl 过表达突变体,以及由 35SCaMV 启动子驱动表达 OsICL 的 WT 过表达突变体。在盐胁迫下,OsICL 表达系的萌发率以及幼苗鲜重和干重均高于 Aticl 突变体,而具有 icl 突变体背景的两条线与 WT 相似。OsICL 表达系的 F/F 和莲座叶的温度受盐胁迫的影响小于 Aticl 突变体。最后,盐胁迫下 Aticl 突变体的葡萄糖和果糖含量最高,而 OsICL 表达系的含量与 WT 相似或低于 WT。
作为一个盐响应基因,OsICL 在转基因拟南芥系中得到了表征,揭示了 OsICL 的表达可以逆转 Aticl 敲除突变体的盐敏感性表型。这项工作提供了新的证据,支持 ICL 通过乙醛酸循环在植物耐盐性中的作用,以及 OsCam1-1 可能参与调节其转录。