| Sorghum-sudangrass is a typical forage crop with heterosis.However,the molecular mechanism of heterosis is still unknown.In this study,we used the method of transcriptome analysis to identify the differentially expressed genes in order to analyze the molecular mechanism of heterosis formation and to accumulate data for the study of heterosis genetic theory.The results are as follows:1.With 4 sorghum male sterile lines and 5 types of sudan grass as parents,according to NC II design prepared into 20 hybrid combinations.The agronomic traits of the hybrids and their parents were measured,and the 8 dominant combinations were selected through the analysis of the phenotypic value and heterobeltiosis.The cDNA-AFLP differential display technique was used for the analysis.(1)12 pairs of primers amplified 315 bands of TDFs,the expression of genes between hybrids and their parental types are:the expression of a single type(P1F1 type)and type two(P2F1),the specific expression of hybrid type(F1 type),unp type(PI type)and type two(type P2),parents were silent type(P1P2 type)and the expression of hybrid parent type(P1F1P2 type)seven.(2)In the display type difference and correlation analysis of yield components,effective tiller number and fresh weight per plant P1F1 type,type and P1P2 type,leaf length and P2 was significantly positively related to phase into plant leaf number and F1 type has significant negative correlation.There was a significant positive correlation between P2 and P1,P1F1P2 and P1P2,and there was a significant negative correlation between leaf width and leaf type.In the correlation analysis with heterosis,ear length was positively correlated with P2F1 type,and leaf width was negatively correlated with P2F1 and P1P2.(3)The differential display types P1F1,P2F1,PI and P2 are dominant effects,accounting for 91.4%of the total detections.The difference display type F1 and P1P2 showed super-dominant,accounting for 4.8%of the total detection,indicating that the heterosis of each trait was mainly affected by(super)dominant effect.(4)The 8 TDFs related to the heterosis of the hybrid were obtained by homologous analysis and BLAST analysis.It was found that these proteins play an important role in the control of plant growth and development.(5)The nucleotide sequences of the eight differential fragments obtained by cloning were verified by semi-quantitative RT-PCR.2.Further selection of strong heterosis of 11A X white shell Sultan grass and its parents,RNA-seq high-throughput sequencing methods were used to the material of root and stem and leaf of three kinds of tissue were analyzed for each sample set of three biological replicates.(1)Sequencing produces 787386707 reads,197.36 Gb the Clean Data,GC content is between 56.08%?58.77%,the sample Q30 base percentage not less than 93.43%.69.32%?85.51%of the samples were compared with the reference genome.(2)The samples were detected with no less than 58,000 SNP sites.After screening,198 SNPs were obtained.Among the 198 SNPs,79,53 and 66(involving 58,38 and 49 transcripts)of root,stem and leaf showed SNPs that showed allele expression bias.There are 10 SNPs(involving 8 transcripts)expressed in all three tissues.(3)A total of 1300 new genes discovered,there are 776 new genes than to known database.Among them,there are 67 new genetic comparisons to the COG database;443 new genes than to GO in the database;120 new genes than to KEGG database;273 new genes than to Swiss-Prot database;769 new genetic comparisons to the nr database.(4)The differentially expressed genes of the hybrid and the female parent 11A were higher In all the differentially expressed genes,and the proportion in root,stem and leaf tissues were 93%,77%and 74%,respectively.3954 high quality differentially expressed genes are divided into 12 types.The expression patterns of most of the differentially expressed genes in the hybrids and their parents were non additive,and the additive gene was about 5.77%and the dominant gene was 0.83%.Respectively using Blast2 GO and KOBAS(v2.0)software to analyze 3455 dominant DEGs,get 46 function subclasses and 83 metabolic pathways.Further analysis revealed that 217 dominant DEGs were expressed in all three tissues,and the 217 genes were analyzed by KEGG and GO.KEGG analysis showed that there are three pathways(Carbon metabolism,Citrate cycle and Glycolysis/Gluconeogenesis)were related to 7 gene.GO analysis showed that nuclear outer membrane and ADP binding were significantly enriched,involving 13 genes.(5)6 genes were selected from the above 20 genes for qRT-PCR verification. |
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