Cloning And Preliminary Functional Analysis Of Melon CmHB?-1 And CmFER10

Melon?Cucumis melo L.?is an important dicotyledonous crop and economic crop.It is a year-round oyster or climbing herb that is widely cultivated all over the world.In this study,two differential expressed genes which are related to fruit development,CmHD-Zip??Homeodomain Leucine Zipper?Family transcription factorsand and CmFER10?FERONIA receptor-like kinase?were chosen from the transcriptome analysis in the melon?Cucumis melo L.cv Hetao?.Functional analysis of its cloning and fruit development.The main results are as follows:?1?Nine members of HD-Zip II family and fifteen members of FER family were identified in melon genome with bioinformatics methods.Meanwhile,Chromosomal location revealed that CmHBII-1 and CmFER10?FERONIA receptor-like kinase?genes were located on chromosome 1 and 10 of melon,respectively.Promoter analysis revealed CmFER10 Contain Ethylene-responsive element,Participation in Stress Response G-boxelement,ABA-responsive element ABRE.?2?The expression pattern of CmHB?-1 and CmFER10 were analysed in the different tissiue and development stages by RT-qPCR.The results indicated that the two genes were differencely expressed in fruit development.The expression of CmHBII-1 gene increased during fruit climacteric stages.The highest expression of CmFER10 genes in fruit growth period.?3?The cDNA of CmHB?-1 and CmFER10 genes were cloned by RT-PCR.And overexpression vectors and gene editing vectors were constructed respectively.The transformed seeds of T₁ generation were obtained by ovary injection and the plant positive rates were detected.Overexpression of CmHB,Gene Editing of CmHB,Overexpression of CmFER and Gene Editing of CmFER were average12.6%,11.1%,15.5%,13.3%,respectively.Seeds of highest positive rate in import melon were selected randomly sown in solar greenhouse.DNA positive test was extracted from tender leaves and self-pollination.Through phenotype observation of T₁ generation melon,it was found that the genetic transformation plants with overexpression of CmHBII-1 were obvious premature phenotype and average earlier4.5 days than normal melon.The quantitative PCR showed that the expression of ACO1 was 12 times higher than common melon.It is indicated that HBII-1 has a regulatory role in melon fruit ripening.The genetic transformation plants with gene editing of CmHBII-1 were Late-maturing phenotype and average late 3 days than normal melon.The genetic transformation plants with overexpression of CmFER were premature phenotype and earlier 2 days than normal melon.