| Nelumbo nucifera Gaertn is the only aquatic flower among the top ten traditional flowers in China.It has very important edible,ornamental and medicinal value.In the long-term natural selection and long history of cultivation,the lotus has a variety of varieties.In order to make full use of the lotus germplasm resources,various morphological and DNA level features have been applied to the related research of lotus germplasm resources,but the research purposes are biased and the conclusions are different.Due to the lack of lotus germplasm resources of Yunnan and the full utilization of lotus germplasm resources,Firstly,this study used 52 N.nucifera accessions planted in Kunming as materials,using phenological observation and morphological observation methods to analyze its adaptability in Kunming.Secondly,94 lotus materials were used as research materials,The genetic diversity and genetic relationship of 94 lotus materials were analyzed and compared using InDel,EST-SSR and SSR molecular markers.The main results were as follows:Through the adaptive observation of the introduction of 52 N.nucifera accessions in Kunming,it was explored that all varieties can grow in Kunming.Most of the varieties have good adaptability in Kunming,only 8 varieties have poor adaptation,which means no flowering,further domestication observations are needed.The research results show that the lotus has strong adaptability and provides a good foundation for its promotion and application in Kunming.26 excellent varieties such as 'Yayuhong No.11','Juwuba','Shennvzi',and so on;18 better varieties such as' Yanlongyeyue','Dazuhonglian' and 'Longfei',and so on were selected;And 8varieties with poor adaptability in the first year: 'Jinhongpao','Qianbanlian','Zhuyishizhe','Taoranyixiao','Hongtailian','qiuyu','Jianwu ','Molingqiuse'.(2)66 alleles were detected by 20 pairs of InDel primers,the average gene diversity index was 0.5233,and the polymorphism information content was between0.2213-0.7381,with an average of 0.4369.It indicates that the genetic diversity of the lotus variety resources revealed by the InDel marker is abundant.The genetic similarity coefficient between the tested materials was 0.1579-1,with an average of 0.5448.The results showed that the tested materials included materials with large genetic differences,but also materials with small genetic differences and rich genetic diversity.Cluster analysis showed that the tested materials were divided into 10 groups.The results of cluster analysis were significantly correlated with provenance,petaltype and plant type,but not with flower color.`Tieling-001` is a single group;tropical lotus and its hybrids are clustered in N.nucifera Gaertn strains,some of which are clustered in one sub-branch;Seed Lotus Series is clustered in N.nucifera Gaertn,but clustered in different sub-branches with flower lotus;SN-001 from Japan is clustered in a single group,and N.nucifera Gaertn is clustered in different sub-branches;N.lutea Pers is clustered in a single group.(3)79 alleles were detected by 20 pairs of EST-SSR primers,the average gene diversity index was 0.5696,and the polymorphism information content was between0.3518-0.4921,with an average of 0.4921.It indicated that the genetic diversity of the lotus variety resources revealed by EST-SSR markers was abundant.The genetic similarity coefficient between the tested materials ranged from0.1633 to 1,with an average of 0.529.The results showed that the tested materials included materials with large genetic differences,but also materials with small genetic differences and rich genetic diversity.Cluster analysis showed that the tested materials were divided into 10 groups.The results of cluster analysis were significantly correlated with provenance,petal type and plant type,but not related with flower color.‘N.lutea Pers ' and N.nucifera Gaertn were clustered into one branch,some of them were clustered into N.nucifera Gaertn,and some of them were only one branch,which indicated that there was a great genetic difference between N.lutea Pers and N.nucifera Gaertn,and no obvious rule was found in the clustering of other materials.(4)88 alleles were detected by 20 pairs of SSR primers,with an average gene diversity index of 0.5872 and polymorphism information content of 0.0982-0.7645,with an average of 0.5235.The results showed that the genetic diversity of lotus cultivars revealed by SSR markers was rich.The genetic similarity coefficient between the tested materials ranged from0.2456 to 0.9535,with an average of 0.5048.The results showed that the genetic diversity of the tested materials was abundant.Cluster analysis showed that most of the petal type,plant type and provenance were clustered into one group,but a few of them were clustered into different types of groups,which indicated that the clustering results were closely related to petal type,plant type and provenance.‘N.lutea Pers' and some Chinese-American hybrid lotus are clustered into one branch,and some Chinese-American hybrid lotus are classified into N.nucifera Gaertn groups;‘Fenhonlingxiao' comes from Thailand is belongs to tropical lotus,and ‘Fenhonlingxiao'ddis clustered into a single group in thisclustering system;Seed lotus series are clustered in N.nucifera Gaertn,but there is a clear distinction between them.(5)Combining the molecular data of InDel,EST-SSR and SSR,the genetic similarity coefficients of the three markers were analyzed.The similarity coefficients between InDel and EST-SSR were 0.3941,between EST-SSR and SSR were 0.4937,and between InDel and SSR were 0.3847.The correlation between the three markers was poor.The correlation coefficients between the combined InDel,EST-SSR and SSR marker data and the similarity coefficient matrices of InDel,EST-SSR and SSR are 0.7662,0.7894 and 0.7967,respectively.The correlation between the merged data and the three markers was better,and the order of correlation was SSR > EST-SSR >InDel.Clustering analysis of combined data showed that most of the same phenotypic materials can be clustered together,and a few of them can be clustered in different phenotypic groups.‘N.lutea Pers' is separately clustered into one group;some Chinese-American hybrid lotus is clustered into one group,some Chinese-American hybrid lotus is clustered into N.nucifera Gaertn,'fenhonglingxiao' and N.nucifera Gaertn are clustered into one group,and 'SN-001' and N.nucifera Gaertn are clustered together,and only one sub-branch.Seed lotus series is clustered in N.nucifera Gaertn,but there is a clear distinction between them.(6)Comparing the results of cluster analysis and genetic diversity analysis of four types of markers,the efficiency of detection results of four types of markers was as follows: combining three types of markers > SSR > EST-SSR > InDel. |
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