盐渍化对甜瓜生长的影响及甜瓜耐盐相关基因CmRAV1的初步研究

发布时间：2018-06-16 10:56

本文选题：甜瓜 + 盐胁迫　；参考：《上海交通大学》2015年硕士论文

【摘要】：甜瓜(Cucumis melo L.)属于葫芦科植物,是我国重要的经济栽培作物。土壤盐渍化会严重影响甜瓜的生长发育,降低产量及品质。本文以课题组前期筛选的两个甜瓜品种(‘冰雪脆’为耐盐;‘玉露’为盐敏感)为材料,研究盐渍化对甜瓜生长的影响,并运用分子生物学手段开展甜瓜CmRAV1基因的初步研究。主要结果如下:本论文首先是在实验室前期甜瓜耐盐种质资源筛选的基础上,利用耐盐品种‘冰雪脆’和盐敏感品种‘玉露’,同时利用生物炭对盐渍化土壤进行改良,研究生物炭处理对土壤和甜瓜生长的影响。结果表明,生物炭处理明显降低了土壤中可溶性盐分含量以及土壤呼吸作用;同时提高了有机质含量及植株的净光合速率,但是对甜瓜生物量及果实品质没有显著差异。300mM NaCl处理后,两个甜瓜品种植株地上部和地下部鲜重均有所下降。但是,品种‘冰雪脆’鲜重无论是地上部还是地下部,均高于品种‘玉露’。随着盐处理时间的延长,两品种叶绿素含量呈现下降趋势,直至24h后,相比于对照差异显著。盐处理对甜瓜根茎叶中离子含量的影响表现为:Na+在茎中的积累量最大,叶片次之,根中最少。同时K+含量在盐处理后根中显著降低,茎中变化不明显,叶片中显著增加。CmRAV1基因是从实验室前期的研究(盐胁迫下两个甜瓜品种耐盐转录组测序结果及盐胁迫下构建的cDNA文库)中筛选出来的,属于AP2/ERF基因家族的RAV亚家族。序列分析表明,CmRAV1包含1个AP2结构域和1个B3结构域,全长1182bp,编码321个氨基酸。序列比对发现,CmRAV1与黄瓜中一个RAV同源性最高。CmRAV1基因受盐胁迫诱导表达,同时该基因在甜瓜根部及花中的表达量相对较高。将CmRAV1的ORF序列连接至植物表达载体PHB-YFP中,通过注射烟草表皮细胞研究该基因在细胞中的定位情况。结果显示该基因主要在细胞核中表达。实验通过酵母表达载体pYeDP60,将CmRAV1转化到酵母菌株G19中。结果表明,CmRAV1可以提高酵母细胞的耐盐性。
[Abstract]:Cucumis melo L. Belongs to Cucurbitaceae plants, is an important economic cultivation crops in China. Soil salinization will seriously affect the growth and development of melon and reduce yield and quality. In this paper, the effects of salinization on the growth of muskmelon (CmRAV1 gene) were studied by means of molecular biology. The main results are as follows: firstly, on the basis of the screening of salt-tolerant germplasm resources of muskmelon in the early stage of laboratory, the salt-tolerant varieties "ice and snow brittle" and salt-sensitive variety 'Yulu' were used to improve the salinized soil at the same time. The effects of biochar treatment on soil and melon growth were studied. The results showed that the content of soluble salt in soil and soil respiration were significantly decreased, and the content of organic matter and the net photosynthetic rate of plants were also increased by biochar treatment. However, there was no significant difference in biomass and fruit quality of muskmelon. However, the fresh weight of the variety was higher than that of the variety Yulu. With the prolongation of salt treatment time, the chlorophyll content of the two varieties decreased, and after 24 hours, there was a significant difference compared with the control. The effect of salt treatment on ion content in rhizome leaves of muskmelon showed that the accumulation of Na in stems was the largest, followed by leaves and roots. At the same time, the K content decreased significantly in the roots after salt treatment, but the change in the stem was not obvious. The significant increase of CmRAV1 gene in leaves was isolated from the pre-laboratory studies (sequence analysis of salt tolerant transcriptome and cDNA library constructed under salt stress), which belongs to the RAV subfamily of AP2 / ERF gene family. Sequence analysis showed that CmRAV1 contained an AP2 domain and a B3 domain, with a total length of 1182 BP, encoding 321 amino acids. Sequence alignment showed that CmRAV1 had the highest homology with a single RAV in cucumber. CmRAV1 gene was induced to express under salt stress, and the expression of CmRAV1 gene was relatively high in the roots and flowers of muskmelon. The ORF sequence of CmRAV1 was ligated into the plant expression vector PHB-YFP, and the localization of the gene was studied by injecting tobacco epidermal cells. The results showed that the gene was mainly expressed in the nucleus. CmRAV1 was transformed into yeast strain G19 by yeast expression vector pYeDP60. The results showed that CmRAV1 could improve the salt tolerance of yeast cells.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S652;S156.41

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