Cucumber (Cucumis sativus L.) is one of the most widely cultivated vegetables in the world. High temperature and other stress conditions can affect the growth and development of this plant, even leading to the decreases in yield and quality. The mitogen-activated protein kinase (MAPK) family plays a crucial role in plant stress responses. However, the role of MPK4 in the stress response of cucumber remains to be reported. In this study, we cloned CsMPK4, which encoded 383 amino acid residues. The qRT-PCR results showed that the expression level of CsMPK4 was the highest in leaves and flowers, moderate in roots, and the lowest in stems and tendrils. CsMPK4 was located in the nucleus and cytoplasm, and it had a close relationship with CmMPK4 in muskmelon. The cucumber plants overexpressing CsMPK4 became stronger and shorter, with reduced length and quantity of tendrils. Moreover, the transgenic seedlings were more resistant to high temperatures, with decreased malondialdehyde (MDA) content and increased activities of peroxidase (POD) and superoxide dismutase (SOD) in young leaves. Furthermore, the protein-protein interaction between CsMPK4 and CsVQ10, a member of the valine-glutamine family, was confirmed by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. The results suggested that CsVQ10 cooperated with CsMPK4 in response to the high temperature stress in cucumber. This study laid a foundation for the further study on the stress response mechanism of CsMPK4 and the breeding of stress-resistant cucumber varieties.
Cucumis sativus/metabolism* ; Mitogen-Activated Protein Kinases/physiology* ; Plant Proteins/metabolism* ; Plants, Genetically Modified/metabolism* ; Gene Expression Regulation, Plant ; Stress, Physiological/genetics* ; Cloning, Molecular

Cucumber (Cucumis sativus) is an important vegetable crop in the world. Agrobacterium-mediated transgenic technology is an important way to study plant gene functions and improve varieties. In order to further accelerate the transgenic research and breeding process of cucumber, we described the progress and problems of Agrobacterium tumefaciens-mediated transgenic cucumber, from the influencing factors of cucumber regeneration ability, genetic transformation conditions and various additives in the process. We prospected for improving the genetic transformation efficiency and safety selection markers of cucumber, and hoped to provide reference for the research of cucumber resistance breeding and quality improvement.
Agrobacterium tumefaciens ; metabolism ; Breeding ; Cucumis sativus ; genetics ; microbiology ; Plants, Genetically Modified ; microbiology ; Research ; Transformation, Genetic

The replicase genes of five isolates of Cucumber green mottle mosaic virus from Jiangsu, Zhejiang, Hunan and Beijing were amplificated, sequenced and analyzed. The similarities of nucleotide acid sequences indicated that 129 kD and 57 kD replicase genes of CGMMV-No. 1, CGMMV-No. 2, CGMMV-No. 3, CGMMV-No. 4 and CGMMV-No. 5 were 99.64% and 99.74%, respectively. The similarities of 129 kD and 57 kD replicase genes of CGMMV-No. 1, CGMMV-No. 3 and CGMMV-No. 4 were 99.95% and 99.94%, while they were lower between CGMMV-No. 2 and the rest of four reference sequences, just from 99.16% to 99.27% and from 99.04% to 99.18%. All reference sequences could be divided into six groups in neighbor-joining (NJ) phylogenetic trees based on the replicase gene sequences of 129 kD, 57 kD protein respectively. CGMMV-No. 1, CGMMV-No. 3 and CGMMV-No. 4 were clustered together with Shandong isolate (Accession No. KJ754195) in two NJ trees; CGMMV-No. 5 was clustered together with Liaoning isolate (Accession No. EF611826) in two NJ trees; CGMMV-No. 2 was clustered together with Korea watermelon isolate (Accession No. AF417242) in phylogenetic tree of 129 kD replicase gene of CGMMV; Interestingly, CGMMV-No. 2 was classified as a independent group in phylogenetic tree of 57 kD replicase gene of CGMMV. There were no significant hydrophobic and highly coiled coil regions on 129 kD and 57 kD proteins of tested CGMMV isolates. Except 129 kD protein of CGMMV-No. 4, the rest were unstable protein. The number of transmembrane helical segments (TMHs) of 129 kD protein of CGMMV-No. 1, CGMMV-No. 2, CGMMV-No. 3 and CGMMV-No. 5 were 6, 6, 2 and 4, respectively, which were 13, 13 and 5 on the 57 kD protein of CGMMV-No. 2, CGMMV-No. 4 and CGMMV-No. 5. The glycosylation site of 129 kD protein of tested CGMMV isolates were 2, 4, 4, 4 and 4, and that of 57 kD protein were 2, 5, 2, 5 and 2. There were difference between the disorders, globulins, phosphorylation sites and B cell antigen epitopes of 129 kD and 57 kD proteins of tested CGMMV isolates. The current results that there was no significant difference between the replicase gene sequences, it was stable and conservative for intra-species and clearly difference for inter-species. CGMMV-No. 1, CGMMV-No. 3, CGMMV-No. 4 and CGMMV-No. 5 had. a close genetic relationship with Shandong and Liangning isolates (Accession No. KJ754195 and EF611826), they are potentially originate from the same source. CGMMV-No. 2 was closer with Korea isolate. High sequence similarity of tested samples were gathered for a class in phylogenetic tree. It didn't show regularity of the bioinformatics analysis results of 129 kD and 57 kD proteins of tested CGMMV isolates. There was no corresponding relationship among the molecular phylogeny and the bioinformatics analysis of the tested CGMMV isolates.
Computational Biology ; Cucumis sativus ; chemistry ; classification ; enzymology ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Diseases ; virology ; RNA Replicase ; chemistry ; genetics ; metabolism ; Sequence Homology, Nucleic Acid ; Viral Proteins ; chemistry ; genetics ; metabolism

The effects of heavy metal cadmium (Cd), alone and in combination with zinc (Zn), on the root growth and activity of antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD) in Cucumis sativus L. hairy roots were studied. The purpose was to study the possibilities on using C. sativus hairy roots for phytoremediation of cadmium contamination. The results showed that less than 10 mg/L Cd enhanced the growth of C. sativus hairy roots and increased root diameter only in 5-15 days of root culture. At Cd concentrations above 15 mg/L hairy root growth was gradually inhibited with increasing Cd concentration. The roots formed were shorter with smaller lateral roots. Among all the Cd concentrations tested, except with 10 mg/L Cd, the soluble protein contents in the C. sativus hairy roots cultured with the other Cd concentrations decreased, but the POD and SOD activities increased gradually with time during the culture process. Further tests were conducted using a control culture containing 25 mg/L Zn alone. The addition of 1 mg/L Cd to the 25 mg/L Zn culture stimulated the growth of C. sativus hairy roots after 7-15 days of growth, compared with the control. At all other Cd concentrations the growth of C. sativus hairy roots was inhibited compared to the control. Growth inhibition increased with increasing Cd concentration, and the hairy roots formed fewer, shorter and smaller lateral roots, the tips of which became swollen. After 5 days culture with different concentrations of Cd + 25 mg/L Zn, the root biomass and the activity of POD and SOD were lower than in C. sativus hairy roots cultured without the addition of Zn. However, the soluble protein content was significantly higher when the culture contained 25 mg/L Zn. Our results suggested that C. sativus hairy roots have higher tolerance to heavy metal Cd but higher concentration of Cd inhibited the growth. Cd in combination with Zn would result in more serious Cd-induced growth inhibition.
Biodegradation, Environmental ; Cadmium ; pharmacology ; Cucumis sativus ; enzymology ; growth & development ; Peroxidase ; metabolism ; Plant Roots ; enzymology ; growth & development ; Superoxide Dismutase ; metabolism ; Tissue Culture Techniques ; Zinc ; pharmacology

Effects of phosphorus deficiency in the medium on growth and morphology and activities of SOD and POD, utilization of nitrogen and calcium in the medium during liquid culture of Cucumis sativus hairy roots were investigated. The results showed that C. sativus hairy roots can not grow in the medium without addition of any phosphorus. When cultured into the medium with different Pi concentrations, the growth of C. sativus hairy roots was significantly inhibited with the decreasing of Pi concentration in the medium, its main roots became thinner and longer, the number of its lateral roots was decreased and its lateral roots became shorter and smaller. Compared to the medium with full strength phosphorus, the content of soluble proteins in C. sativus hairy roots cultured under Pi deficiency was significantly lower than that with standard full-length Pi, while POD and SOD activities in C. sativus hairy roots were significantly stimulated. Compared to the control (without addition of any phosphorus in the medium), the activities of POD and SOD in C. sativus hairy roots cultured in the medium with different concentration of Pi were lower than the control. When C. sativus hairy roots were cultured into medium with different Pi concentrations, the medium conductivity was gradually decreased with time and with direct proportion of the initial Pi concentration of the medium; NH4(+)-N and NO3(-)-N of the medium was gradually absorbed and utilized, at day 15, NH4(+)-N of the medium was nearly used up but its NO3(-)-N was not used up until cultured for 30 days. Pi deficiency in the medium could decrease the consumption rate of NO3(-)-N and inhibited the absorption and utilization of calcium of the medium by C. sativus hairy roots. Proper enhancement of Pi concentration could stimulate absorption and consumption of calcium of the medium.
Calcium ; metabolism ; Cucumis sativus ; growth & development ; metabolism ; Culture Media ; Nitrogen ; metabolism ; Phosphates ; pharmacology ; Plant Roots ; growth & development ; metabolism ; Superoxide Dismutase ; metabolism ; Tissue Culture Techniques