Cloning,Expression And Functional Analysis Of BZIP11 And FaSigma Transcription Factors From Strawberry

Light is the most important environmental factor for plant growth and development in nature.It can not only regulate the growth and development of plants,but also participate in photosynthesis and affect the accumulation of plant photosynthesis products.The previous study found that some bZIP and Sigma family transcription factors are involved in the development of plant organs and fruits.The FabZIP11 and FaSigma transcription factors obtained by cloning in this experiment may be related to the transportation and distribution of photocontracted products in strawberries based on the related genes of previous studies.Therefore,to explore the functions of FabZIP11 and FaSigma transcription factors,it is possible to explore the mechanism of photosynthetic product distribution in the growth and development of strawberry organs and fruits,which has important theoretical basis and application value in improving strawberry yield and quality.In this paper,the fruits of Benihoppe strawberry were used as experimental materials,and two transcription factors,FabZIP11 and FaSigma.were cloned according to the results of the integrated analysis of the previous transcriptome and metabolome.Bioinformatics analysis and function prediction were preformed to identify the gene function of these two transcription factors.The expression patterns of FabZIP11 and FaSigma in different parts of stems of different varieties of strawberry and leaves of Benihoppe,Akihiime and Sweet Charlie at different development stages were analyzed by real-time quantitative PCR.At the same time,the sugar,acid and chlorophyll contents of fruits,leaves and stems in different parts of three strawberry varieties were detected.Construction of overexpression vectors pCXSN-Flag-FabZIP11 and pCXSN-Flag-FaSigma,pCXSN-Flag-FaSigma successfully transformed Arabidopsis Col-0,and identified several Arabidopsis transgenic plants;construction and construction of overexpression vector PB1121-gfp-FabZIP11 and PB1121-gfp-FaSigma,and transformed several tomato seedlings,and injected tomato and strawberry fruits for functional verification.The above experiments preliminarily verified the gene function of FabZIP11 and FaSigma?This experiment achieved the following main results:1.Using Benihoppe fruit as experimental material,a bZIP and a Sigma transcription factor were cloned.The results of the phylogenetic tree showed that2.FabZIP11 and AtbZIP11 had the highest homology,so they were named FabZIP11,and another gene had a close homology with grape RNA polymerase transcription factors,and was named FaSigma.2.Temporal and spatial expression analysis of FabZIP11 and FaSigma in fruits,leaves and stems of three strawberry varieties of Benihoppe,Akihiime and Sweet Charlie at different development stages,found that FabZIP11 and FaSigma genes showed both in leaf gro fruit ripening process the obvious downward trend,only FaSigma gene in Benihoppe leaf showed an upward trend.The transcript level of FabZIP11 and FaSigma had higher expression pattern in the stolonous near the mother plant.3.The relationship between the physiological indexes of sugar,acid and other physiological indexes of three strawberry fruits and leaves at different development stages of Benihoppe,Akihiime and Sweet Charlie and the chlorophyll content was detected.As a result,it was found that the content of soluble solids continued to increase as the fruit matured.During the development of leaves and fruits,the content of soluble total sugar is also constantly accumulating,and the content of titratable acid is also rising.The chlorophyll content shows a decreasing trend during the fruit development and an increasing trend during the leaf development.The sugar,acid,and content of stolons in different parts are very low,and there is no obvious change trend.Chlorophyll is the content of stolon away from the mother plant is much higher than the stem near the plant.4.The overexpression vector pCXSN-Flag-FaSigma was constructed,and the heterologous plant Arabidopsis thaliana was successfully transformed.Finally,11 transgenic plants were obtained.The overexpression vectors PBI121-gfp-FabZIP11 and PBI121-gfp-FaSigma were constructed,and the heterologous plant tomato was successfully transformed,and 10 transgenic tomato seedlings were obtained.The FabZIP11 and FaSigma genes were overexpressed in tomato and octoploid strawberry fruits by injection,and fruit ripening was significantly inhibited in the injection area.RT-PCR results found that the expression of FabZIP11 and FaSigma genes in the injection area increased by 8-100 times and 6-20 times,indicating that FabZIP11 and FaSigma genes have the function of inhibiting the ripening of tomatoes and strawberries.