| The composition and content of wheat gluten determine the processing quality of wheat flour,and wheat gliadin is one of the important components of gluten,accounting for about 50% of its total.Gliadin mainly determines the viscosity and ductility of wheat gluten.In gliadin,?-gliadin has a significant correlation with the quality of wheat.In this study,the high-quality strong gluten wheat variety Zhengmai 379 was used as the material to clone the ?-gliadin family gene and predict the antigenic site of celiac disease gene.The transcriptome technology was used to calculate the cloned gene in 120 natural samples.The expression level in the population material,combined with the determination results of its quality indicators,used correlation analysis to analyze the effect of ?-gliadin gene on various quality indicators of wheat,and based on transcriptome sequencing technology,the correlation analysis of m RNA sequence expression level was carried out.The results of the study are as follows:(1)Using degenerate PCR technology,130 ?-gliadin genes were cloned from Zhengmai 379,including 50 gene sequences with complete open reading frames,analyzing the structural features of amino acid sequences and constructing phylogenetic tree findings.The complete gene sequence contains 8 conserved cysteine residues with typical ?-gliadin structural features.The sequences Gli-?-001 to Gli-?-008 have identical amino acids encoded at other positions except for the same cysteine positions.The results of phylogenetic tree analysis were divided into four branches.The I branch belongs to the novel ?-gliadin multi-gene subfamily.The antigenic site analysis was performed in the cloned ?-gliadin by bioinformatics method.10 antigenic sites,the class I amino acid sequence contains 0 to 1 Celiac disease(CD)antigenic site,and other types of sequences of celiac antigenic sites are more concentrated and concentrated.(2)By statistically analyzing the quality indexes of 120 natural group materials,it is found that the flour whiteness,gluten and silty quality indicators are basically the same except that the flour formation time,stabilization time and silty quality index are skewed.The tendency to form a normal distribution.Among these indicators,except for the negative correlation between the weakening degree and other indicators,other indicators have a significant positive correlation at the 0.01 level,especially the correlation between the silty quality index and the formation time and stabilization time(r value respectively is 0.85,0.90).(3)Using transcriptome sequencing technology,the 130 ?-gliadin gene sequences cloned in this experiment were used as reference genes to calculate the correlation between m RNA expression levels and quality indicators.The analysis showed that the transcriptional expression levels of Gli-?-074,Gli-?-075,Gli-?-076 and Gli-?-109 were significantly higher than other genes,and the expression levels of these four m RNAs were related to silty,wet gluten and pull.The correlation between the PMT and A3 values related to the extensibility index was low,but it was significantly positively correlated with the weakening degree.The results showed that the ?-gliadin gene had a negative effect on the wheat gluten strength.(4)According to the m RNA expression of ?-gliadin gene,56 gene sequences with RPKM value greater than 1 were selected and analyzed by genome-wide association using a mixed linear model.The analysis found that the key sites controlling the expression levels of m RNA sequences of Gli-?-004,Gli-?-110,Gli-?-111 and Gli-?-124 are consistent,which are mainly distributed on chromosomes 1A,1B and 1D,among which More distributed on chromosome 1A.On chromosomes 1A,1B and 1D,and the effect value of controlling the m RNA expression level locus is significantly higher,the phenotypic contribution rate of the single associated locus is between 25.3 and 58.73. |
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