Study On Genes Related To Flavonoids Synthesis Of Nitrogen Ion Implantation On Trollius Chinensis

Trollius chinensis is mostly a wild resource.In recent years,with the continuous exploration of its medicinal and ornamental value,the demand for Trollius chinensis has increased greatly,resulting in serious damage to the wild Trollius chinensis resources.Therefore,it is of great research value to study the yield,quality and environmental adaptation of Trollius chinensis,reveal the relevant ways to synthesize flavonoids,design efficient molecular improvement breeding strategies,and create excellent strains of Trollius chinensis with high yield,high quality and wide adaptability.In this study,the seeds of Trollius chinensis were irradiated with different doses of nitrogen ions.It was found that the germination rate and germination potential of seeds were increased to a certain extent by lower radiation dose.When the dose reached 7,8×10¹⁵N⁺/cm²,the germination rate decreased seriously,which decreased by 45.71%and55.36%compared with the control.The plant height of Trollius chinensis has the same rule.Low dose can promote the growth of Trollius chinensis,but when the dose is too high,the growth of Trollius chinensis is inhibited.After nitrogen ion irradiation,the content of flavonoids in leaves increased in different degrees,and when the injection dose was 1×10¹⁵N⁺/cm²,the content of flavonoids was the highest,which was more than twice that of the control group.After irradiation,Trollius chinensis also produced various variations in flower shape and leaf shape,such as sepals becoming more orderly and compact,leaves being more slender or shorter and more compact.The transcriptome of Trollius chinensis leaves was sequenced by high-throughput sequencing technique,and finally 36 101 Unigene were obtained by mixed assembly.The N50 length was 1 789 bp,and the average length reached 1 204 bp;.A variety of enzyme genes and transcription factors involved in the synthesis of flavonoids,such as chalcone synthase and isoflavone 7-O-methyltransferase,were found Six candidate reference genes were mined from Trollius chinensis transcriptome database,and the stability of gene expression was evaluated.It was found that the combination of reference genes stably expressed by Trollius chinensis in different doses of nitrogen ion implantation,different developmental stages and different tissues was ACT and CYP,TUA and ACT,UBQ and CYP,respectively.The expression levels of flavonoids synthesis related genes in leaves of the first dose treatment group and the control group were studied.It was found that there was a high positive correlation between the expression levels of CHI and CHS genes and flavonoids content,a significant positive correlation between b HLH gene expression and flavonoids content,and a significant negative correlation between MYB gene expression and flavonoids content.The main innovations of this paper are as follows:1.By annotating and analyzing the transcriptome data of Trollius chinensis leaves,the metabolic pathway map of flavonoids in Trollius chinensis was drawn.2.Identification of internal reference genes suitable for quantitative expression of Trollius chinensis.