Metabolomics Analysis Of Thin-shell Fagopyrum Tataricum Seeds And Gene Cloning And Expression Of AGL11

Thin shell tartary buckwheat has the characteristics of thin shell and easy shelling,which provides valuable germplasm resources for raw and processed tartary buckwheat.In this experiment,guimi tartary buckwheat No.18(M18)was used as the main material to study the agronomic and yield characteristics,seed phenotype characteristics,main storage content,GC-MS metabolic component analysis,and AGL11 gene cloning and expression analysis.The main results are as follows:1.The results showed that the cell layers of M18 shell were less than that of JQ2,and there was a clear gap between the shell and the kernel.Phenotypic analysis showed that M18 seeds were shorter than JQ2,without obvious edge,and some seeds had broken shell.The shell rate of M18 was 14.99%,which was significantly lower than that of JQ2.2.The yield per plant of M18 was 13.73%lower than that of JQ2,reaching a significant difference level;but the kernel weight per plant was not significantly different from that of the control.At the same time,the number of grains per plant of M18 was 46.58%higher than that of JQ2,reaching a very significant difference level.The amylopectin and amylose of M18 were 39.49%and 15.94%respectively,which were slightly lower than those of JQ2,but did not reach the significant level.The glutenin,albumin and globulin of M18 were 1.37%,3.10%and 0.19%respectively,which were significantly or extremely lower than those of JQ2.3.By GC-MS,92 kinds of metabolites were identified in the seeds of M18,XMQ and JQ2,including organic acids(31%),amino acids(24%),sugars(11%),polyols(7%),phosphoric acids(7%),fatty acids(3%),amines(2%)and others(15%).Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(opls-da)showed that M18,XMQ and JQ2 could be distinguished well in the early,middle and late stages of grouting,indicating that the metabolites of M18,XMQ and JQ2 were different in the early,middle and late stages of grouting;in addition,M18 and XMQ,M18 and JQ2,JQ2 and XMQ could be distinguished well in the early,middle and late stages of grouting,indicating that they could be distinguished well in each grouting stage There are differences in metabolites among varieties.Volcanic map analysis showed that M18,XMQ and JQ2 were up-regulated in the early and middle stages of grouting,and amino acids were concentrated in the middle stage of grouting.At the later stage of filling,the contents of erythritol,galactose,glucose,isomaltose,aspartic acid,glutamic acid and tryptophan in M18 were significantly different from those in JQ2;the contents of arabinose,catechin,citric acid,galactose,glucose,alanine,ribose and isomaltose in XMQ were significantly different from those in JQ2.The contents of galactose and glucose in M18 and XMQ were higher than JQ2.The metabolic pathways involved in these distinct metabolites include tRNA biosynthesis,alanine aspartate glutamate metabolism,glycine serine threonine metabolism,citric acid cycle,nitrogen metabolism,glycolysis and other metabolic pathways.4.AGL11 was cloned from M18,M55,M104,CMQ and JQ2 seeds.The full-length sequence of AGL11 gene is 672 bp,encoding 220 amino acids.It is predicted that AGL11 is an unstable hydrophilic protein,and the subcellular location of AGL11 is in the nucleus or mitochondria.Cluster analysis showed that JQ2 formed a big branch,M18 formed a big branch with M104,M55 and CMQ,and JQ2 of thick shell buckwheat was obviously separated from thin shell buckwheat.Quantitative analysis showed that the expression of AGL11 gene was the highest in thin shell tartary buckwheat M18,XMQ and thick shell tartary buckwheat JQ2 at ovule development stage,slightly decreased in early filling stage,and the lowest in late filling stage.In conclusion,the differences of seed shell morphology between M18 and JQ2 of were confirmed in this experiment.AGL11 sequence was cloned.It was confirmed that AGL11 gene expression in thin shell and thick shell tartary buckwheat is significantly different.92 kinds of metabolites were detected by GC-MS.Metabonomics analysis showed that many metabolites in sugar metabolism and citric acid circulation pathway of thin shell tartary buckwheat were up-regulated and many metabolites in amino acid metabolism pathway were down regulated.The results of yield and quality analysis showed that the weight of M18 1000 grains was lower,but the number of grains per plant was significantly increased.The starch and total flavonoids of M18 seeds were similar to JQ2.These results can provide scientific reference for further exploring the molecular regulation mechanism of tartary buckwheat seed and shell development and high-yield breeding of hybrid thin shell tartary buckwheat.