Constitutive expression of CaRma1H1, a hot pepper ER-localized RING E3 ubiquitin ligase, increases tolerance to drought and salt stresses in transgenic tomato plants

CaRma1H1, an endoplasmic reticulum (ER)-localized hot pepper really interesting new genes (RING) E3 Ub ligase, was previously reported to be a positive regulator of drought stress responses. To address the possibility that CaRma1H1 can be used to improve tolerance to abiotic stress in crop plants, CaRma1H1 was constitutively expressed in transgenic tomato (Solanum lycopersicum) plants. CaRma1H1-overexpressing tomato plants (35S:CaRma1H1) exhibited greatly enhanced tolerance to high-salinity treatments compared with wild-type plants. Leaf chlorophyll and proline contents in CaRma1H1 overexpressors were 4.3- to 8.5-fold and 1.2- to 1.5-fold higher, respectively, than in wild-type plants after 300?mM NaCl treatment. Transgenic cotyledons developed and their roots elongated in the presence of NaCl up to 200?mM. In addition, 35S:CaRma1H1 lines were markedly more tolerant to severe drought stress than were wild-type plants. Detached leaves of CaRma1H1 overexpressors preserved water more efficiently than did wild-type leaves during a rapid dehydration process. The ER chaperone genes LePDIL1, LeBIP1, and LeCNX1 were markedly up-regulated in 35S:CaRma1H1 tomatoes compared with wild-type plants. Therefore, overexpression of CaRma1H1 may enhance tomato plant ER responses to drought stress by effectively removing nonfunctional ubiquitinated proteins. Collectively, constitutive expression of CaRma1H1 in tomatoes conferrred strongly enhanced tolerance to salt- and water-stress. This raises the possibility that CaRma1H1 may be useful for developing abiotic stress-tolerant tomato plants. Key message CaRma1H1 increases drought tolerance in transgenic tomato plants.     

过表达TaLEA1和TaLEA2基因提高转基因拟南芥的耐盐性

我国土壤盐碱化日益严重,对我国的粮食安全造成了严重威胁。耐盐基因挖掘对作物耐盐育种非常重要。LEA蛋白家族是一个多基因家族,在植物应对非生物胁迫中发挥重要作用。本课题组前期研究阐明小麦TaLEA1基因在拟南芥中过表达可以提高转基因植物的耐盐性和抗旱性。本研究系统分析了小麦TaLEA2基因表达蛋白的理化性质、基因表达模式及启动子功能区域,并在拟南芥中过表达TaLEA2基因及共表达TaLEA1和TaLEA2基因,分析TaLEA2基因的抗逆功能及2个LEA基因的抗逆效果。结果表明,TaLEA2基因的表达产物属于第3组LEA蛋白,是稳定的亲水蛋白,富含α-螺旋、β-转角等结构。TaLEA2基因在小麦根、茎、叶、花、种子等不同组织中均有表达,盐胁迫条件诱导其高表达。在拟南芥中过表达TaLEA2基因,或过表达TaLEA1和TaLEA2基因都能够提高转基因拟南芥的耐盐性和抗旱性,转基因株系的种子萌发率、根长及叶绿素含量显著高于野生型,且双基因过表达的转基因植物的抗逆能力高于单个基因过表达株系。本研究结果为LEA基因抗逆机理的研究和多基因共转提高植物抗逆性提供了重要信息。     

Arabidopsis mutants deregulated in RCI2A expression reveal new signaling pathways in abiotic stress responses

To uncover new pathways involved in low-temperature signal transduction, we screened for mutants altered in cold-induced expression of RCI2A, an Arabidopsis gene that is not a member of the CBF/DREB1 regulon and is induced not only by low temperature but also by abscisic acid (ABA), dehydration (DH) and NaCl. This was accomplished by generating a line of Arabidopsis carrying a transgene consisting of the RCI2A promoter fused to the firefly luciferase coding sequence. A number of mutants showing low or high RCI2A expression in response to low temperature were identified. These mutants also displayed deregulated RCI2A expression in response to ABA, DH or NaCl. Interestingly, however, they were not altered in stress-induced expression of RD29A, a CBF/DREB1-target gene, suggesting that the mutations affect signaling intermediates of CBF/DREB1-independent regulatory pathways. Several mutants showed alterations in their tolerance to freezing, DH or salt stress, as well as in their ABA sensitivity, which indicates that the signaling intermediates defined by the corresponding mutations play an important role in Arabidopsis tolerance to abiotic stresses. Based on the mutants identified, we discuss the involvement of CBF/DREB1-independent pathways in modulating stress signaling.