Developement Of Molecular Markers For High Carotenoid Content In Foxtail Millet

Foxtail millet is one of our country's first cultivation crops,it has cold and drought resistance,long storage time,high yield and other fine features.Removal of the lemma is called millet,millet rich in vitamins,essential amino acids,mineral elements,fatty acids and other nutrients,and easily absorbed by the body.the main component of millet yellow pigment is carotenoids,including ?-carotene and lutein,they have antioxidant and enhanced immune function.In this study,60 different foxtail millet from different ecological regions in China were cultivated by two years of field experiment.The agronomic traits of millet were analyzed,and the varieties with high carotenoid content and low content were screened to hybridization to construct their F2 isolates.High performance liquid chromatography(HPLC)and liquid chromatography mass spectrometry(LC-MS)were used to select the optimum conditions for the determination of ?-carotene and lutein in foxtail millet.The highest and lowest varieties of total content of beta-carotene and lutein were screened out from 60 different millet varieties,take the middle and the late grouting samples of these two varieties from the field.RNA was extracted and sequenced.The experiment consisted of two replicates in the middle stage and three replicates in the latter stage,a total of 10 samples were selected.Through the bioinformatics analysis method,the design of the comparison program to screen out the different expression of genes,digging high carotenoid related molecular markers.The main findings are as follows:(1)The four agronomic traits plant height,ear length,single ear weight and 1000 grain weight of 60 varieties were determined,the average plant height of the 60 varieties of millet was 146.14 cm,the average panicle length was 20.77 cm,the average panicle weight was 23.7 g,and the average 1000 grain weight was 3.42 g.(2)The carotenoids were extracted from the foxtail millet and the contents of ?-carotene and lutein were determined by HPLC and LC-MS.The highest and lowest varieties of?-carotene and lutein total content were No.52 baimigu and No.43heizhigu,respectively.(3)The correlation between four agronomic traits and ?-carotene and lutein in 60 varieties of millet was analyzed.Among them,1000-grain weight was not significantly correlated with other traits and carotenoid content,and the single spike has significant relevance at 0.01 level with lutein content,plant height,spike length,There was a significant correlation between the plant height,panicle length and the content of lutein at the 0.5 level.(4)Design comparison program(52 mid VS52 late,52 mid VS43 mid,52 late VS43 late,43 mid VS43 late).Through matching sequences we find the intersection between them,based on the comments on the differentially expressed genes,6367 differences were found and identified,among them,there has 40 genes to be related to carotenoid synthesis.(5)According to the sequencing analysis,32 SSR markers related to carotenoid synthesis were found.There are five types of SSR markers,among which the P1 SSR markers is the most,followed is P2,P3 and c,and the number of c*is the least.