| Melon(Cucumis melo L.) is an important economic crop cultivation. It belongs to the family of Cucurbitaceae. Soil salinization will seriously affect the growth and development of melon. It can can inhibit the growth, affect metabolism, reduce yield and quality of melon. Now, two melon varieties(’Bingxuecui’ for salt; ’Yulu’ for salt-sensitive) based on Our previous research were used to study the effects of saline soil on melon growth. Meanwhile, the work of Cm RAV1 gene cloning, family analysis, subcellular localization and functional were performed by the technology of molecular biology. The following were research results:In order to explore the effects of biochar on saline soil improvement and crop growth, the two melon(Cucumis melo L.) varieties ‘Bingxuecui’ and ‘Yulu’ was carried out under saline soil with organic fertilizer treatment(Control) and with biochar treatment. Our results demonstrated that biochar can reduce soil respiration, soluble salt content, greenhouse gas CO2 emissions and improve the net photosynthetic rate, fruit weight and quality. But it plays little role on melon’s biomass and fruit.After 300 m M Na Cl treatment, the shoot and root fresh weight of two melon varieties were declined. However, no matter aboveground or underground part, the fresh weight of ‘bingxuecui’ was higher than the cultivar ’Yulu’. With the extension of the salt treatment time, chlorophyll SPAD values of two varieties always has been showing a downward trend. After 24 h, it appeared significant difference compared with control. The ion contents in melon root, stem leaf were different after salt treatment. Na + accumulation in the stem is the largest, leaf followed, at least in the root. And K + content was decreased significantly in the root, changed mildly in stem and increased significantly in the leaves.In this study, we cloned a melon RAV gene, Cm RAV1, based on our previous transcriptome sequencing assay, which was performed under salt stress. Cm RAV1 contains an AP2 domains and a B3 domains, it belongs to the AP2/ERF family, RAV subfamily. The full-length c DNA of Cm RAV1 contained 1182 nucleotides with an open reading frame(ORF) of 963 nucleotides. Sequence alignment was found that Cm RAV1 has a higher homology with a predicted cucumber RAV. RT-PCR was performed to investigate the regulation of Cm RAV1 expression by salt stress. Our results showed that the transcript levels of Cm RAV1 are induced by Na Cl. Additionally, tissue expression showed that Cm RAV1 m RNA accumulates higher in root and flowers.In this experiment, we constructed a recombinant plant expression plasmid by fusing the ORF sequence of Cm RAV1 with a fluorescence protein YFP. Then, it was transferred into the tobacco epidermal cells. Fluorescence emissing from the expressed product localizing was visualized using laser-scaning confocal microscope techniques. The results show that the Cm RAV1 gene was localized in the nucleus.In order to investigate the function of the protein encoded by Cm RAV1, we introduced and heterologously expressed Cm RAV1 c DNA in yeast cells and analyzed their growth. Yeast expressing Cm RAV1 gene grew much better on medium containing Na Cl than did the control vector. This results show that Cm RAV1 can enhanced the tolerance of the yeast cells to high-salinity. |
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