Transcriptome Sequencing And Differential Expression Gene Analysis Of Grain Grouting In No-Waxy And Waxy Foxtail Millets

Waxy endosperm influence cooking and eating quality of millet.According GBSSI,SSS and SBE activity dynamics changes,in grouting,immature grouting grains of waxy millet Gonggu68(grouting period 1 and 5 days)and non-waxy millet Chigu4(grouting period 1 and 10 days)were used to analyze their transcriptome sequences by using Illumina Hiseq4000.The results showed that:1.The activities of three enzymes showed a low-high-low trend,and the activities of GBSSI,SSS and SBE reached the maximum on the 5th,10th and 3rd day of grouting.2.Homology alignment information from unigene annotation to the eggNOG and KEGG database were 73752 and 37785,respectively.The unigene can be divided into 24 categories according to eggNOG functions.The unigene sequences were annotated to organic system,metabolism,genetic information processing,environmental information processing and cell process,associated with 2457,20382,10214,2259 and 2473 unigene,respectivety.345137 unigene were divided into three main bodies of biological processes,cellular components and molecular functions according to GO functional classification,which containing 25,15 and 10 GO subcategories,respectively.3.When the highest and lowest GBSSI activity,the number of differentially expressed genes in waxy millet was less,and the number of up-regulated genes was 431 more than down-regulated genes;the number of differentially expressed genes in non-waxy millet was higher,and the number of up-regulated genes was 97 more than down-regulated genes.It proved that the number of differential genes in the process of grain formation in the waxy millet was not changed,and the up-regulated gene was the dominant factor.The number of differential genes in the process of grain formation in the non-waxy millet changed greatly.4.In grouting early-middle stage,obviously GO enrichment of differentially expressed genes in waxy type were biogenetic function of seed oil bodies,but GO enrichment of differentially expressed genes in non-waxy type were Photosynthesis,light capture I in optical system and pigment binding functions in.In grouting early-middle stage,obviously GO enrichment of differentially expressed genes in no-waxy and waxy types were aromatic compound biosynthetic process.5.The main pathway of the differentially expressed in gene no-waxy and waxy types were plant hormone signal transduction in KEGG enrichment;The plant vegetation-pathogen interaction pathways were significant performance in no-waxy type,but not waxy type.6.Ten differential genes with high expression levels(TPM values)and large differences were screened out.The differential genes SSII-3 and PHO1 were down-regulated in A2 in the comparison group A2-VS-A1.Up-regulated expression in A1;differential gene AS is up-regulated in A2 and down-regulated in A1.In the comparison group B2-VS-B1,the differential genes PHO1-1,AGP 16 and WAXY were down-regulated in B2 and up-regulated in B1;the differential gene AS was up-regulated in B2 and down-regulated in B1.In the comparison group B1-VS-A1,the differential gene GT7 was up-regulated in B1 and down-regulated in A1;the differential gene CESA1 was up-regulated in B1 and down-regulated in A1.In the comparison group B2-VS-A2,the differential gene CESA7 was down-regulated in B2 and up-regulated in B1.7.Using Actin(Si001873)as an internal reference gene,10 differentially expressed genes related to waxy and no-waxy were verified by qRT-PCR,which was consistent with the transcriptome results.It was proved that the 10 differential genes selected were indeed related to glutinous rice.