Full
Outline
4-coumarate CoA ligase（4CL）is a key enzyme in the phenylpropane pathway controlling the biosynthesis of lignan, lignin and other compounds closely related to Niubang（Arctium lappa）microscopic characterization of stone cell development and Niubang（Arctium lappa）glycoside accumulation, the aim of this study was to identify the 4CL genes in Niubang（Arctium lappa）and explore the biological functions of Niubang（Arctium lappa）4CL enzyme gene family＇s biological functions.
In this study, it cloned and sequenced 8 predicted 4CL gene coding sequences（CDS）in the burdock genome, performed bioinformatics analysis on the obtained sequences and performed prokaryotic expression of Al4CL3.
The eight members of Niubang（Arctium lappa）4CL gene family were identified and cloned with 100% consistency with the predicted sequences, except for Al4CL5（96%）and Al4CL8, which had five variant sites. Al4CL3, Al4CL5 and Al4CL8 proteins had two transmembrane helical structures; subcellular localization in the cytoplasm, plasma membrane, chloroplast and endoplasmic reticulum and gene structure. There were some differences in the gene structure, unevenly distributed on chromosomes 1, 2, 3, 9, 10 and 14, and there was a 4CL amino acid conserved domain in the amino acid sequence. There were multiple cis-acting elements in the promoter region of the Al4CL gene, which was induced by multiple hormones or stresses. The phylogenetic analysis showed that Al4CL was divided into three subfamilies, and the covariance analysis showed that there was a closer relationship between members of the Al4CL family of genes and the prickly bracteate thistle. There was a closer relationship between members of the Al4CL gene family and the prickly bracteate thistle. Recombinant plasmid pET-28a-chAl4CL3 was constructed, successfully induced the expression of chAl4CL3 protein and purified.
The excavation and characterization of burdock 4CL gene family provides a certain theoretical basis for the biosynthesis of Niubang（Arctium lappa）and its quality improvement, and provides a reference for the in-depth study of Al4CL and its biological functions.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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