As the group II LEA (late embryogenesis abundant) proteins, dehydrins (DHNs) play an important role in plant growth and development, as well as in response to abiotic or biotic stress challenges. In this study, a DHN gene named CsLEA11 was identified and characterized from Cucumis sativus. Sequence analysis of CsLEA11 showed that it is a Y3SK2-type DHN protein rich in hydrophilic amino acids. Expression analyses revealed that the transcription of CsLEA11 could be significantly induced by heat and cold stress. The recombinant plasmid was transformed into Escherichia coli BL21 and isopropy-β-d-thiogalactoside (IPTG) was used to induce recombinant E. coli to express CsLEA11 gene. Overexpression of CsLEA11 in E. coli enhanced cell viability and conferred tolerance to heat and cold stress. Furthermore, CsLEA11 protein could protect the activity of lactate dehydrogenase (LDH) under heat stress. Taken together, our data demonstrate that CsLEA11 might function in tolerance of cucumber to heat and cold stress.

Stress-associated proteins (SAPs) are a class of zinc finger proteins that confer tolerance to a variety of abiotic and biotic stresses in diverse plant species. However, in cucumber (Cucumis sativus L.), very little is known about the roles of SAP gene family members in regulating plant growth, development, and stress responses. In this study, a total of 12 SAP genes (named as CsSAP1-CsSAP12) were identified in the cucumber genome, which were unevenly distributed on six chromosomes. Gene duplication analysis detected one tandem duplication and two segmental duplication events. Phylogenetic analysis of SAP proteins from cucumber and other plants suggested that they could be divided into seven groups (sub-families), and proteins in the same group generally had the same arrangement of AN1 (ZnF-AN1) and A20 (ZnF-A20) domains. Most of the CsSAP genes were intronless and harbored a number of stress- and hormone-responsive cis-elements in their promoter regions. Tissue expression analysis showed that the CsSAP genes had a broad spectrum of expression in different tissues, and some of them displayed remarkable alteration in expression during fruit development. RT-qPCR results indicated that all the selected CsSAP genes displayed transcriptional responses to cold, drought, and salt stresses. These results enable the first comprehensive description of the SAP gene family in cucumber and lay a solid foundation for future research on the biological functions of CsSAP genes.

Germins and germin-like proteins (GLPs) are glycoproteins closely associated with plant development and stress response in the plant kingdom. Here, we carried out genome-wide identification and expression analysis of the GLP gene family in cucumber to study their possible functions. A total of 38 GLP genes were identified in cucumber, which could be mapped to six out of the seven cucumber chromosomes. A phylogenetic analysis of the GLP members from cucumber, Arabidopsis and rice showed that these GLPs could be divided into six groups, and cucumber GLPs in the same group had highly similar conserved motif distribution and gene structure. Gene duplication analysis revealed that six cucumber GLP genes were located in the segmental duplication regions of cucumber chromosomes, while 14 genes were associated with tandem duplications. Tissue expression profiles of cucumber GLP genes showed that many genes were preferentially expressed in specific tissues. In addition, some cucumber GLP genes were differentially expressed under salt, drought and ABA treatments, as well as under DM inoculation. Our results provide important information for the functional identification of GLP genes in the growth, development and stress response of cucumber.

Catalase (CAT) is a key scavenging enzyme for the degradation of hydrogen peroxide (H₂O₂) and plays an important role in the tolerance to diverse abiotic stresses in many different organisms. In this study, we characterized the function of a catalase gene (CsCAT3) previously isolated from Cucumis sativus in the defence against a variety of stresses. Protein alignment and phylogenetic analysis revealed that CsCAT3 was clustered in the dicot group and shared 41%–95% identity with other plant CATs. Expression analyses revealed that the expression of CsCAT3 was induced by diverse abiotic stresses such as heat, polyethylene glycol, cold and NaCl treatment, as well as by signalling molecules such as abscisic acid (ABA) and H₂O₂. An Escherichia coli heterologous expression system was constructed to characterize the function of CsCAT3 in vitro. Its overexpression in E. coli could increase the tolerance to heat, cold, salinity and osmotic conditions. These results indicate that CsCAT3 plays important roles in abiotic stress tolerance and that CsCAT3 confers tolerance in E. coli recombinants against abiotic stresses, which may be due to the increased activities of antioxidant enzymes that reduce the oxidative damage caused by stress conditions.

The growth regulating factor (GRF) family is a conserved class of transcription factors involved in various biological processes in plants. However, there have been only a few studies of the GRF family genes in cucumber, Cucumis sativus (Cs). In this study, we identified and characterized 8 CsGRF genes in cucumber. Two highly conserved domains, QLQ and WRC, were identified to be present in all CsGRF proteins. In addition, three less conserved domains (FFD, TQL, and GGPL) were also detected in some CsGRF members. Based on phylogenetic analysis, the GRF genes from cucumber, Arabidopsis, tomato, rice and maize could be classified into 10 groups, and CsGRFs were clustered closer with the GRF genes from dicots (Arabidopsis and tomato) than with those from monocots (rice and maize). Promoter analysis revealed that the CsGRF genes were involved in cucumber growth and development as well as in responses to various hormones and stresses. Transcriptome data showed that the CsGRF genes have distinct expression patterns in different tissues, especially in ovaries and leaves. Expression profiling analysis indicated that all CsGRF genes were responsive to salt and drought stress treatments. These results demonstrate that the cucumber GRF gene family may function in organ development and plant stress responses.

The AGL15 subfamily MADS-box proteins play vital roles in various developmental processes, such as floral transition, somatic embryogenesis, and leaf and fruit development. In this work, an AtAGL15 ortholog, CsMADS26, was cloned from cucumber (Cucumis sativus L.). The open reading frame (ORF) of CsMADS26 is 669 bp in length, encoding a predicted protein of 222 amino acids. The CsMADS26 protein contains a highly conserved MADS-box domain and a variable C domain, as well as less conserved I and K domains. Phylogenetic relationship analysis revealed that CsMADS26 was clustered into the AGL15 clade of AGL15 subfamily. Expression analysis based on qRT-PCR showed that CsMADS26 is mainly expressed in reproductive organs including flowers and fruits. Transgenic Arabidopsis plants with ectopic expression of CsMADS26 exhibited curled rosette and cauline leaves, and the leaf size was much smaller than that of wild-type (WT) plants. These results provide clues for the functional characterization of CsMADS26 in the future.