Cloning And Analysis Of Resistant Functions Of Transcription Factors ZmWRKY58from Zea Mays

Drought and high-salinity are two main abiotic stresses which can adversely affect the growth, development and productivity of plants. For the development of sustainable agriculture, it is important to study the mechanisms of plant response to environmental stresses and breed some new crop cultivars with better stress tolerance. Many studies have indicated that transcription factors play a very important role in plant growth and tolerance to environmental stresses. WRKY transcription factor has become one of the hot spots in the gene function research. WRKY transcription factors are known to function in plant growth and development, as well as in adaption to biotic and abiotic stress. In this study, we isolated a WRKY gene, ZmWRKY58, from maize inbred line B73and then we analyzed its functions of stress tolerance. The main results as follows:1. The results of Real-time PCR showed that ZmWRKY58was induced by PEG and Nacl.2. This study successfully cloned the ZmWRKY58gene by PCR method. The nucleotide sequence total length is1110bp which containing only one conserved WRKY domain with a zinc finger motif of C2H2. ZmWRKY58encode369amino acids of the coding region, suggesting its relative molecular mass is39.4kD, and isoelectric point is9.81.3. To confirm ZmWRKY58was a transcription factor and indeed localized to the nucleus, a plant expression vector p1302-ZmWRKY58was constructed and the fusion protein was bombarded into the onion epidermal cells. The results indicated that ZmWRKY58was localized into the nucleus.4. An overexpression vector CPB-ZmWRKY58was constructed and was transformed into rice calli by Agrobacterium-mediated method. Five T0overexpression rice lines were obtained. Three independent T3transgenic homozygous lines L1-2, L1-3, L4-5were investigated in the forthcoming experiment. A double-stranded RNA interfering vector CPB-2ZmWRKY58i was also constructed.5. Physiology analysis research proved that MDA content and relative electrical conductivity of transgentic ZmWRKY58rice was lower WT. But the survival rate and relative water content was hihger than WT. The results suggested that ZmWRKY58gene confered salt and drought tolerance in transgenic rice.6. According to root elongation experiment on ABA medium, ZmWRKY58was found to be more sensitive in transgenic rice. The results suggested that ZmWRKY58may be involved in regulating plant growth and development processes mediated by ABA.