玫瑰 RrWRKY47X1-2 基因克隆及耐盐功能验证

Cloning and Salt Tolerance Function Verification of Gene RrWRKY47X1-2from Rosa rugosa

摘要

土壤盐渍化是制约植物生长的主要非生物胁迫之一。本研究以玫瑰（ Rosa rugosa）为材料，对玫瑰 RrWRKY47X1-2基因的耐盐功能进行研究。该基因编码的蛋白质含有440个氨基酸，属于WRKY Group IIb亚家族，定位于细胞核且具有转录激活活性。200 mM NaCl胁迫下 RrWRKY47X1-2响应强烈，在玫瑰叶片中呈双峰表达模式（峰值4 h和72 h）。通过在本氏烟草中异源过表达该基因，结果发现与野生型相比，过表达株系（OE47）叶片萎蔫程度轻于野生型（WT），叶片中丙二醛（MDA）含量降低，超氧化物歧化酶（SOD）活性和可溶性蛋白含量显著升高，盐胁迫相关基因表达上调。沉默玫瑰 RrWRKY47X1-2 后，植株盐敏感性增强，盐胁迫相关基因表达受到抑制。结果表明， RrWRKY47X1-2（GenBank登录号：PX533116）可能通过调控抗氧化系统、渗透平衡及激素通路正向调节植物耐盐性，研究可为玫瑰耐盐育种提供候选基因。

Abstract

Soil salinization is one of the main abiotic stresses that restrict plant growth. This study uses Rosa rugosa as a material to investigate the salt tolerance function of the RrWRKY47X1-2 gene in roses. The protein encoded by this gene contains 440 amino acids and belongs to the WRKY Group IIb. It is located in the nucleus and has transcriptional activation activity. Under 200 mM NaCl treatment, RrWRKY47X1-2exhibits a strong response, with a bimodal expression pattern in rose leaves (peaking at 4 h and 72 h). By heterologous overexpression of this gene in Nicotiana benthamiana, it is found that compared to the wild type, the overexpressed line (OE47) has less leaf wilting than the wild type (WT), reducing levels of malondialdehyde (MDA), significantly increasing superoxide dismutase (SOD) activity and soluble protein content, and upregulating expression of salt stress-related genes. Virus-induced gene silencing (VIGS) of RrWRKY47X1-2 in R.rugosa increases the plant's salt sensitivity and suppresses the expression of salt stress-related genes. The results indicate that RrWRKY47X1-2(GenBank accession number: PX533116) may positively regulate salt tolerance by modulating the antioxidant system, osmotic balance, and hormone pathways, providing a candidate gene for salt-tolerant rose breeding.

关键词

玫瑰 / RrWRKY47X1-2 / 基因过表达 / 基因沉默 / 盐胁迫

Key words

Rosa rugosa / RrWRKY47X1-2 / gene overexpression / gene silencing / salt stress

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基金资助

山东省重点研发计划项目“林草种质资源精准鉴定与创新利用”(2024LZGC003)

国家自然科学基金委员会，青年科学基金项目（C类）(32501683)