Study On The Expression Of Carotenoids Synthesis Related Genes In Yellow-fleshed Apple

Apple fruit colour and lustre are the key indicators of commercial evaluation and the fruit nutrition health care function, fruit colour and lustre is mainly composed of carotenoids, chlorophyll, anthocyanin pigment, flavonoids and other relative content of the decision. The study of carotenoids in Apple is less,weak foundation, and carotenoids for apple quality is important, therefore carotenoid synthesis for apple related gene expression studies, will provide important theoretical basis for breeding of high quality.This article use the color-difference meter measuring the apple color value iduring the period of development, carotenoids and other skin pigments content are measured with colorimetric method,analyzing the change of the skin pigment, color difference and its correlation; Through the adoption of a variety of apple test materials, a variety of methods for RNA extraction and quality inspection, select the RNA extraction method suitable for the transcriptome sequencing of apple fruit; Turn to ‘dongbeihuang crabapple’ color period before and after the fruit of the transcriptome sequencing, and validate the transcriptome data, analyze the fruit development of mesocarp and fruit meat carotene synthesis related gene expression, screen the carotenoid metabolism significant genes with the carotenoid content change. the main results and conclusions are as follows:Yellow germplasm ‘dongbeihuang crabapple’, Red germplasm ‘donghongguo’ and White germplasm‘ wanbai crabapple’ development of mesocarp carotenoids, chlorophyll, total phenol and flavonoids downward trend on the whole, carotenoids and chlorophyll in ‘dongbeihuang crabapple’nearly mature content increased slightly; Carotenoids and chlorophyll a and chlorophyll b, total chlorophyll, anthocyanins, flavonoids, total phenol and L * values are significant or extremely significant correlation; Relative to red germplasm ‘donghongguo’ and White germplasm‘wanbai crabapple’, Yellow germplasm ‘dongbeihuang crabapple’ carotenoid content is higher, about22.73μg/g,and in Yellow germplasm carotenoids has weak correlation with other pigments.Apple leaves and different fruits as material, adopt seven different method of4 kinds of materials to extracted RNA, and by gel electrophoresis, Agilent 2100 RNA chip inspect the quality of total RNA extraction. Results show that there are two kinds of methods can successfully extract the RNA from 4four kinds of materials, it is concluded that modified CTAB method is suitable for the transcriptome sequencing of apple fruit RNA extraction.Transcriptome sequencing for Yellow germplasm ‘dongbeihuang crabapple’color period before and after fruit, through data analysis can known a total of 3056 differentially expressed genes, turn color after fruit has 1270 genes in higher expression, 1786 genes were expressed. Turn color after cut the most significant genes involved in photosynthesis, and cysteine, methionine metabolism and styrene acrylic pigment synthesis genes increase quantity is more, involving carotenoids synthetic route has seven differentially expressed genes. Verifies the fluorescence quantitative PCR with two periods of apple fruit resources ‘dongbeihuang crabapple’, ‘jilinxiaohuang crabapple’ and ‘huayue’, and detection three species fruit carotenoids content in the two periods, comprehensive analysis MdPSY2\MdCCD4a\MdCCD4b genes differentially expressed genes in four has the most significant influence for carotenoid content; To ‘dongbeihuang crabapple’, and ‘wanbai crabapple’ 20 DAFB\50 DAFB\90DAFB\mature fruit peel as a material for the transcriptome data validation, to detect carotenoids synthetic gene expression related to quantity and carotenoid content, and check CCD4 relative expression level in 4 peroids fruits of these crabapple. Comprehensive analysis carotenoid accumulation was significantly associated with MdPSY2、MdCCD4a、MdCCD4b.