| Objective:In this paper,the screening of Camellia nitidissima C.W.Chi excellent germplasm was performed by measuring the content of chemical components with different varieties,and combining the phenotypic traits,biological yield,growth vigor,stress resistance,genetic stability(SSR),and other indicators.At the same time,UPLC fingerprint and SSR fingerprint were constructed to reflect the variety and quantity of C.nitidissima more comprehensively,then to describe and evaluate quality,and to construct the fingerprint of elite germplasm characteristics to determine its superiority,stability,and authenticity.And from the external conditions(soil,climate)and internal germplasm(genetic difference analysis based on SSR molecular markers),the quality forming factors of high-quality C.nitidissima were evaluated,and the quality forming factors were clarified.In order to promote the development of the C.nitidissima industry,it provides a basis for the classification and identification of C.nitidissima varieties and the breeding of fine varieties.Methods:(1)The phenotypic traits of flowers and leaves were observed by visual inspection and tape measure.besides,yield was calculated based on known data,and the growth potential and leaf health were evaluated at the same time.(2)HPLC and UV spectrophotometry were used to determine the chemical components of C.nitidissima,and UPLC fingerprints were constructed;SPSS was used to analyze the content of active components in different types of C.nitidissima,and to evaluate their quality differences;cluster analysis was performed on C.nitidissima to obtain dominant populations;excellent strains were screened in combination with yield and growth potential.(3)Mining SSR molecular markers of C.nitidissima based on transcriptome sequencing technology,designing primers according to the two-end conserved sequences of SSR marker sites,analyzing the polymorphism of the primers to screen out the primers with polymorphism and identify them.Its polymorphism was analyzed.The primers with PIC>0.5 were selected as the core primers to amplify the length of the fragment to measure different kinds of C.nitidissima samples to construct the fingerprints of C.nitidissima varieties and excellent strains.(4)Collecting and measuring soil and climate factors in the field,and using SPSS to analyze the correlation between soil,environment,phenotype,and chemical composition content,to obtain the factors that are significantly related to the content.Results:(1)There were quite differences in 23 golden camellia phenotypic traits,showing rich genetic diversity.Each group after clustering has its own distinct characteristics;there were great differences in yield,growth vigor,and stress resistance among various species.In actual production,it is recommended to select species with high yield for breeding such as Camellia Longzhouensis J.Y.Luo,Camellia Pubipetala Y.Wan et S.Z.Huang,Camellia fusuien S.Y.Liang et X.J.Dong,Camellia chrysantha(Hu)Tuyama var.macrophylla S.L.Mo et S.Z.Huang,Camellia euphlebia Meer.et Seealy,Camellia niridissima Chi,Camellia insularis Orel et Curry or select varieties with high leaf health rate such as Camellia flavida Chang,Camellia limonia f.obovata S.L.Mo et Y.C.Zhong,Camellia tunghinensis Chang.(2)The fingerprints of 30 types of 23 kinds of C.nitidissima were constructed;the dynamic accumulation trend of chemical components suggested that the best sampling time was from the end of August to the end of January of the following year,followed by April and May.The types of chemical constituents in C.chrysantha,C.niridissima,and C.insularis leaves were relatively more than those in flowers,and the contents were quite different.There were also significant differences in the types and contents of chemical components in all C.nitidissima leaves.The highest chemical component content of C.niridissima Chi was 19-nortestosterone,and the lowest was epicatechin.The chemical components of JHCFP1 were relatively rich in variety and high in content;The other types of C.nitidissima tea had the highest content of epicatechin,and the lowest were kaempferol and chlorogenic acid.The chemical composition content of each group of C.insularis was different.The cluster analysis results based on the UPLC control fingerprints of C.nitidissima showed that the first group belonged to most kinds of chemical components,and the content was generally medium to a high level.The second group belonged to more kinds of chemical components,but the content was relatively low.Although there were not many kinds of chemical components in the third group,the content was relatively high;the fourth group belonged to more kinds of chemical components,and the content of the components was at a medium level.(3)Combining the chemical composition content,yield,growth,stress resistance,and other indicators of C.nitidissima,it was concluded that the excellent populations of C.chrysantha were JHCDY1 and JHCDY2,the excellent population of C.euphlebia was JHCXM1,the excellent population of C.niridissima was JHCFP2.The excellent populations of C.insularis were JHCYN4 and JHCYN5.It was further concluded that the excellent strains in C.chrysantha were DY03,DY05,DY06,DY08,DY13,DY16,DY20;the excellent strains in C.euphlebia were XM01,XM02,XM03,XM05,XM10,XM15;The excellent lines in C.niridissima were FP1-47,FP3-90,FP1-48,FP1-50,FP1-46,FP3-73,FP1-35,the excellent lines in C.insularis were YN1-01,YN3-30,YN3-32,YN3-34,YN3-39,YN3-01,YN3-05,YN3-07,YN3-14,combined with phenotypic traits,chemical component content,stress resistance,yield,and other factors to establish C.niridissima and C.insularis variety standards.(4)A total of 32 primers with polymorphic loci were screened from 200 pairs of SSR markers in C.nitidissima.Genetic diversity analysis showed that there was a certain degree of inbreeding rate within the C.nitidissima germplasm,and there was a large degree of inbreeding between the germplasms.32 polymorphic primers were selected from 32 polymorphic primers,30 core primers suitable for C.Niridissima and 29 core primers suitable for C.insularis were screened,and the SSR variety identification map was constructed on this basis.Combined with the chemical composition content,two core primers with significant differences were screened for C.niridissima and C.insularis respectively,and the SSR fingerprint of C.nitidissima excellent strains was constructed at the same time.(5)The results of the correlation analysis of the chemical components of C.nitidissima with soil and climate factors showed that the available phosphorus and available potassium in the soil were significantly correlated with the more chemical components of C.nitidissima,and calcium ions had little effect on the effective components.The rest of the soil factors had a certain significant correlation with the chemical components;among the climatic factors,temperature and humidity,especially humidity,had a greater correlation with the chemical components of C.nitidissima.(6)Through the research on the correlation between chemical components and phenotypic traits of C.nitidissima,it was found that the two phenotypes of leaf tail shape and leaf vein in C.niridissima Chi could be used as phenotypic indicators related to the content of chemical components for breeding;The leaf vein,vein grid and leaf area of C.insularis were considered as phenotypic indicators related to chemical component content in breeding.Conclusion:The similarity evaluation results meet the requirements for establishing liquid phase fingerprints.The quality evaluation results of different kinds of C.nitidissima could effectively screen out the excellent populations and strains of C.nitidissima.Based on SSR molecular marker technology,the genetic diversity of 190 C.nitidissima samples was analyzed.At the same time,core primers were used to construct the SSR variety identification map and the fine germplasm SSR fingerprint map of the samples of C.Niridissima and C.insularis respectively.Based on the correlation analysis of chemical composition content and geographical and environmental factors,the soil and environmental factors affecting the chemical composition content of C.nitidissima were obtained.Based on the correlation analysis of chemical constituent content and phenotypic traits,the phenotypic traits associated with chemical constituents were clarified.The results of this paper could provide a basis for the classification and identification of C.nitidissima varieties,the screening of excellent germplasm,the selection of suitable production environment,and the prediction of high-content germplasm by phenotypic traits. |
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