The Preliminary Functional Analysis Of Melon CmIAA16 And CmGSTU12 Genes In Fruit Development

Melon is a kind of horticultural crop with economic value.It is widely cultivated all over the world due to its nutrient and unique taste.In this study,the Cucumis melo L.cv.Hetao was used as raw material.By analyzing the four stages of fruit growth and development in the melon transcriptome data obtained in our laboratory,two differential expression gene CmIAA16 and CmGSTU12 were obtained.And then bioinformatics analysis,cloning,expression profiling and genetic transformation studies were carried out for them.The main results are as follows:?1?Eighteen Aux/IAA and forty-eight GSTs were identified in melon by two-way comparison method.Analyzing the physicochemical properties of two gene family members such as molecular weight,isoelectric point and number of amino acids,I got analysis and prediction of chromosome localization,gene density,subcellular localization,gene structure,conserved motifs and systemic protein evolution relationships of family members.The results showed that CmIAA16 may regulate the expression of middle and lower genes of the hormone signaling pathway by participating in the corresponding signaling pathways of these hormones,and thus play a role in the growth and development of plants.CmGSTU12 may play a role by participating in hormone induction and responding to abiotic stress.?2?The cDNA of the chosen genes were cloned and the ORF sequences were 714 bp and 657 bp in length.The overexpression vector and gene editing vector were constructed respectively,named pPIAA16,pPGSTU12,pYL-IAA16 and pYL-GSTU12.?3?The real-time fluorescence quantitative method was used to detect the expression of CmIAA16 and CmGSTU12 in different developmental stages of fruits.The results showed that CmIAA16 had the highest expression in the growing period,and the CmGSTU12 had the highest expression in the climacteric stage.In addition,we also verified the functions of these two genes in terms of physiology and biochemistry.?4?The T₁ generation transformed seeds were obtained by using ovary injection method to stably transform the melon.Sixty seeds were randomly selected and tested by PCR.One hundred and twenty seeds were randomly selected from the seed bags with higher positive rate and sown in the greenhouse to observe the relevant phenotypes in the ripening process of T₁ generation fruits.Compared to the control group,the ripening days of T₁ fruits which were transformed pPIAA16 were4.3 days in advance.The results showed that CmIAA16 has the function of promoting fruit ripening,which may be a positive regulator of melon fruit ripening process.However,compared to the control group,the ripening days of T₁ fruits which were transformed pPGSTU12 were 5.5 days later.That is,CmGSTU12 has the function of inhibiting fruit ripening,which may be a negative regulator of melon fruit ripening process.