| Isatis indigotica Fort.is a biennial herb in the Cruciferous family.Its root(isatis indigotica)and leaf(Isatis indigotica)can be used as medicine.It has the effects of clearing heat and detoxifying,cooling blood and removing spots,improving pharynx and relieving pain.As COVID-19 is raging around the world,radix isatidis and its proprietary Chinese medicines play a vital role in prevention and treatment,and their efficacy in preventing and treating influenza viruses and enhancing immunity is increasing the market demand.As its base plant,isatis indigotica has a certain scale in production.In addition,in the growth process of Isatis indigotica,the root of isatis indigotica would be lignified and lose its medicinal value due to premature licorting and flowering in the medicinal material production season of the same year of sowing.Therefore,the problem of premature licorting of Isatis indigotica in the production season became increasingly prominent and gradually became a key problem restricting the production and quality of isatis indigotica.The excellent germplasm resources of medicinal plants are the key and source to ensure the quality of Chinese medicinal materials,and also the material basis for the selection and breeding of excellent varieties of Chinese medicinal materials.So this research through the seed characteristics of 25 I.indigotica germplasm resources,a comprehensive evaluation of medicinal materials and bolting characteristics using grey correlation analysis and DTOPSIS decision,don’t take out moss early screening in medicinal herbs production season of bolting resistance woad germplasm resources,and through the proteomics technology,explore the woad in response to low temperature stress in the process of genetic information related to vernalization,In order to lay a foundation for the subsequent selection of superior varieties of Isatis indigotica,and provide a reference for further excavating the cold tolerance gene of Isatis indigotica and systematically studying the mechanism of cold vernalization.Preliminary results were obtained as follows:1.Screening of bolting-tolerant germplasm of Isatis indigotica.Features of different germplasm woad seed,exuberant growth period,the ground of agronomic traits,medicinal herbs harvest time of the underground parts of shape and quality evaluation index,normal bolting of bolting character indicators before and after anthesis,and through the correlation,the grey correlation degree and DTOPSIS decision analysis methods,such as the woad bolting resistant germplasm screening.The results showed that :(1)there were differences in pod morphology,1000-seed weight,seed germination index and viability among different germplasm of isatis woad.The germination percentage and viability of the germplasm from2016 to 24 were the lowest,which indicated that germination characteristics of isatis indigotica seed had a certain correlation with viability.(2)The aboveground agronomic traits(plant height,plant width,leaf length,leaf width,leaf number per plant and leaf weight)of different germplasm of Isatis indigotica were also different to some extent.The plant height,plant width and leaf width of germplasm 2016-2 were significantly higher than other germplasm,and the germplasm 2016-13 was the smallest or smaller germplasm in leaf width,number of leaves and total weight of leaves.Therefore,it could be considered to eliminate 2016-13 in the later selection process of leaves,and 2016-2 was an excellent germplasm that could be continuously paid attention to.(3)The leaf weight and single root weight of 2016-18 were both the largest.The content of(R,S)-Diyichun in radix isatidis was the highest in 2018-1,and the lowest in 2016-15 and 2016-16.The content of indirubin was the highest in 2016-13,and the lowest in 2016-8.The correlation between the quality of radix isatidis and agronomic traits was analyzed,and the results showed that the root weight and root thick,wide xylem and phloem width and leaf weight and the number of lateral root were very significant positive correlation(P < 0.01),and root weight and leaf extract content and indigo jade red content is negatively correlated,the smaller the yield of radix isatidis,the higher the accumulation of the effective composition content in the leaves.Also from folium isatidis herbs quality analysis results have correlation with the agronomic traits and yield,leaf weight and leaf number,plant,leaf length,xylem width),root diameter and root weight were extremely significant positive correlation(P < 0.01),and negatively correlated with root length and number of lateral root,can speculate when woad underground part accumulated more,the output of the leaf is the lower.There was a significant negative correlation between extract content and root weight(P<0.01),and there was a significant negative correlation between indirubin content and plant height and leaf weight(P<0.05).Therefore,the lower the yield of Leaves was,the higher the content of active components was.In conclusion,according to the requirements of local cultivation conditions and breeding objectives,the type with larger plant width and more leaves per plant can be selected,which is conducive to the breeding of a new strain of isatis indigotica with larger single root weight,so as to achieve the purpose of breeding a new variety of isatis indigotica with high yield.(4)Through the analysis of grey correlation degree and DTOPSIS method,combined with 22 indicators,it was found that the sorting results of grey correlation degree analysis and DTOPSIS analysis were basically consistent with the sorting results of Ci.Finally,the moss-tolerant germplasm of isatis indigotica was screened out from 25 germplasm materials: 2016-4,2016-9,2016-6,2016-15,2016-14,2016-5,2016-1 and 2016-19;The germplasm of medium mossy were2016-26,2016-3,2016-22,2016-18,2016-7,2016-20,2016-25,2016-24 and 2016-21.The easy moss extraction germplasm included 2016-27,2016-16,2016-8,2016-23,2016-2,2016-17,2016-13 and 2018-1.2.Comparative study on photosynthetic characteristics of Isatis indigotica germplasm with different lichens.By comparing the photosynthetic characteristics of moss-tolerant germplasm 2016-5 and moss-easy germplasm 2016-17,the results showed that the net photosynthetic rate of moss-tolerant germplasm 2016-5 was 2.69% higher than that of moss-easy germplasm 2016-17,but the difference was not significant.Stomatal conductance,transpiration rate and intercellular CO2 concentration of the moss-tolerant germplasm were significantly increased by 15.81%,17.22% and 84.80%,respectively,compared with those of the moss-tolerant germplasm.It can be preliminarily indicated that the mossy tolerant germplasm 2016-5 had stronger photosynthetic characteristics compared with the mossy easy germplasm 2016-17.3.Physiological and biochemical studies on different lichen extraction characteristics of Isatis indigotica under low temperature stress.The seedlings of moss-tolerant germplasm 2016-5 and moss-easy germplasm 2016-17 were treated with low temperature stress at different times to measure the changes in physiological and biochemical characteristics.The results showed that: After 4℃ stress at different time,(1)The changes of membrane lipid peroxidation and antioxidant enzyme system of isatis indigotica seedlings showed different trends,and there was no significant difference in SOD activity among different treatments.The activities of CAT and GSH decreased with the extension of stress time,while POD activity increased.(2)The contents of three osmotic substances in isatis indigotica seedlings increased with the extension of stress time,and the increase of soluble sugar content gradually decreased with the extension of stress time,and the content of osmotic substances in moss-tolerant germplasm 2016-5 was higher than that in moss-easy germplasm 2016-17.MDA content of membrane lipid peroxidation products increased with the extension of stress time.(3)The content of endogenous hormone ABA increased and the content of ZT decreased with the prolongation of stress time.The CONTENT of GA in 2016-17 decreased with the extension of stress time,and that in 2016-5 increased with the extension of stress time.4.Transcriptome study of isatis indigotica with different mottling characteristics under low temperature stress.The full-length transcriptome of two isatis indigotica germplasm seedlings with different lichen-extracting characteristics under low temperature stress was obtained by using Nanopore single-molecule real-time sequencing technology.WGCNA was used to identify the transcriptome regulatory network related to soluble sugar,glutathione,proline and zeatin of isatis indigotica germplasm under low temperature.The results showed that: A total of 34329 transcripts were captured after low temperature induction of isatis indigotica,18385 new transcripts were identified,11169 CDS sequences,6168 transcription factors and 30952 SSR types were completely identified,which greatly enriched the data resources of isatis indigotica.Constructed using the identification of transcription of the full transcript of woad cryogenic express network,using 24 of the 15410 sample filtered transcriptional expression module in this building 25,will these modules and woad low temperature induction of soluble sugar,glutathione,proline and zeatin related appraisal to eight Song Lan Chun relevant transcript. |
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