The Morphological Identification And Functional Analysis Of TSK Gene Of Cyathula Officinalis Kuan.

Cyathula officinalis Kuan.is an important medicinal material in sichuan,and the market is mixed with germplasm resources,which seriously affects the safety of clinical medicine.In this experiment,morphological,molecular biology and other techniques were used to identify the tissue morphology of the "BaoXiNo.1" and counterfeit speciesCyathula capitata(Wall.)Moq.,The results showed that the root system was yellowish-brown,and the cells in the root were radially arranged;There are many stem branches,with the growth of acute angle with the main stem,dark red at the stem node,less surface of the surface of the stem,and 4 pulp vascular bundles in the middle of the stem;Leaf is light green,narrow elliptic,the epidermis cells are small and the epidermis is dense,the chloroplast cells are fusiform;The top is a dichasium,with a spike.The Cyathula capitata(Wall.)Moq.is a root system,which is brown,and the cells in the root sheath are arranged in three places;There were few stem branches,with a right angle growth with the main stem,and more surface fur on the stem,and two pulp vascular bundles in the middle of the stem;Leaves in dark green,elliptic,wide leaf epidermis with size irregular macular,epidermal cells is larger and less table fur,rough,chloroplast spherical,a cavity,there are more hungry body autophagy particles;The top is solitary or clustered.At the same time,in view of CoTSK of Cyathula officinalis Kuan.and CcTSK of Cyathula capitata(Wall.)Moq.,and expression vector to construct cloning and bioinformatics analysis,respectively for the subcellular localization of the genes and transgenic arabidopsis preliminary phenotypic restore prokaryotic expression,protein,and modified strain stress response functions such as preliminary explored.The results showed that: the CoTSK gene and the CcTSK gene of the cymbidium,the total length of cDNA were 3855 bp,encoding 1284 amino acids,and there were 7 amino acid differences,and 5'UTR and 3'UTR were different;The systematic evolution analysis of the homologous amino acid sequence of 28 species in the plant community was consistent with the classification rule of single and dicotyledons,which was the most intimate relationship with the ricoid spinach and beet;The quantitative analysis of TSK in Cyathula officinalis Kuan.andCyathula capitata(Wall.)Moq.showed that CoTSK was highly expressed in the stem of Cyathula officinalis Kuan.,and CcTSK was highly expressed in the leaf Cyathula capitata(Wall.)Moq.;Tobacco instantaneous expression verification,CoTSK-GFP and CcTSK-GFP localization of nuclear expression;The root length of the three types of transgenic plants was restored to different degrees in the CoTSK gene;The size of CoTSK and CcTSK prokaryotic protein was approximately 150 kDa;Build into CoTSK/CcTSKgene BL21 strain system,under the stress of DNA damage reagent MMS,number of transgenic strains significantly more than the control.This paper mainly observed the morphological characteristics of Cyathula officinalis Kuan.from cytological level.It can be used effectively for the identification of Cyathula officinalis Kuan.and its counterfeit Cyathula capitata(Wall.)Moq..Secondly,clone CoTSK and explore its initial function,CoTSK and AtTSK have similar functions of regulating plant morphogenesis and responding to DNA damage.In order toprovided reference for the identification of Cyathula officinalis Kuan.and its pseudo Cyathula capitata(Wall.)Moq.,and provide scientific guidance for cultivating high quality Cyathula officinalis Kuan.,at the same time,it will lay a molecular basis for genetic engineering of Cyathula officinalis Kuan..