| Morinda citrifolia, commonly known as NONI, tropical medicinal plants of Rubiaceae genus Morinda. Because of its unique therapeutic and health benefits had became a hot research and development, and develop into a unique high-tech industry in Hainan. There are many research about NONI’s nutrients and pharmacological at home and abroad, the utilization and innovation research of NONI germplasm resources is less. Our research of NONI started relatively late, which seriously hampered the development of NONI industry. In the study of NONI germplasm resources, there is no one using ISSR molecular markers to analysis and research it’s genetic diversity and construct core collection.Using biological traits and ISSR molecular markers to analysis the genetic diversity of the 202 NONI germplasm resource, from the phenotypic and molecular level to reveal genetic diversity and affinity relationships of different germplasm provide the material and theoretical basis for the use of NONI germplasm evaluation, conservation and breeding of new varieties. By comparing three kinds of molecular genetic distance (Sokal-Michener distance, Nei-Lis distance and Jaccard distance), three kinds of sampling methods (clustering-grouping sampling method, stepwise cluster random sampling method and the minimum distance stepwise sampling method), seven kinds of sampling proportion (40%、30%、35%、25%、20%、15%、 10%), combined with biological traits and production practices initially construction NONI’s core collection, then tested and evaluation their genetic diversity. Experiments will improve the efficiency of preservation and the rational use of NONI germplasm. The Core Collection can be put directly into the actual work. The main results are as follows:1.The canopy size, plant height, yield, fruit shape index, leaf index, polysaccharide content, vitamin content, total flavonoid content and coumarin content of 202 NONI were analyzed and evaluated by the correlation analysis, cluster analysis and principal component analysis, preliminary understand genetic diversity of these NONI germplasm.2.There were variation of 202 NONI germplasm between the nine biological traits, coefficient of variation was 8.05-33.61, with an average coefficient of variation was 17.64, the results showed that there are high genetic diversity among NONI germplasm, suitable for constructing core collection. The coefficient of variation of yield, polysaccharides and total flavonoid content were larger, were 33.61,13.00,16.92. They were closely relate to the development and production of NONI, showed that we can improve the yield and economic benefit by selected breeding.3.The result showed the most significant correlation between nine biological traits. Yield with canopy size, height, vitamins, flavonoids, coumarin content was positively correlated. The maximum correlation coefficient between the height and yield was 0.65. The leaf index and polysaccharides, vitamins, flavonoids, coumarin content was negatively correlated.4.Principal component analysis showed that the genetic influence of main ingredient factors in descending order:vitamin, canopy, coumarin, leaf index, plant height, production, fruit shape index, total flavonoids, polysaccharides factor. Contribution rate was closely related to vitamin content and canopy size.5.Cluster analysis showed that:202 NONI mainly divided into three categories, the genetic similarity coefficient ranged from 0.01 to 2.56, showing a highly significant and abundant genetic diversity. Correlation coefficient of clustering result was 0.96886, a better description of clustering results.6.Using the improved CTAB method to obtain a high purity and good quality DNA, meet the needs of ISSR analysis of genetic diversity. Established the optimal reaction of ISSR markers system used Hawaii (17), Indonesia (94), Singapore (193) material of NONI germplasm, optimizedthe main factors affecting temperature, the 13 suitable ISSR primers were screened out.7.13 primers amplified 65 loci, an average of each primer amplified five loci, Among them,63 bands (96.92%) were polymorphic.Variety of qualitative genetic similarity was between 0.55-0.89.NTSYS-pc2.10 software the UPGMA method was applied to cluster 202 NONI species, showed that the genetic similarity coefficient of 0.63, tested materials can be clustered into five groups, showed some characteristics of geographical areas. As standard with Ne, h, I, and the percentage of polymorphic loci (PPB) results showed that the geneticdiversity of germplasm resources of Indonesia and Singapore Morinda citrifolia were high, germplasm in Hainan were relatively low.8.Based on the results of biological traits and molecular marker test, filtered out the best genetic distance of construction core germplasm is the Jaccard.9.1n Jaccard distance, using clustering-grouping sampling method, stepwise cluster random sampling method and minimum distance stepwise sampling method, in accordance with the 7 different sampling proportion (10%,15%,20%,25%,30%, 35%,40%) selected out 21 core germplasm from 202 NONI species. Through the efficacy data (MD:It is traits’ means, SR:tratis’variance, RR:means coincidence rate, CR:means variable rate, PPB:Percentage of significant difference between core collection and the primary collection for percentage of polymorphic loci, MN: Percentage of significant difference between core collection and the primary collection foraverage effective number of alleles, MH: Percentage of significant difference between core collection and the primary collection for average polymorphism information content, MI:Percentage of significant difference between core collection and the primarycollection for average Shannon’s information index) proved the representative of core collection builded by cluster method was better.10.Taking into account biological traits and molecular marker tests result, Tahiti germplasm (177,178) and Haikou, Chang Feng germplasm (194,195) were absorbed into the core germplasm. Core collection of 44 parts (21.8% of the original germplasm), and its Na,Ne, H, I retention rates were 100%,98.4%,97.4%,98.2%; showing that the core collection was a good representative of the original germplasm.11. The NONI core collection included 44 Germplasm, they were:1,9,11,15, 25,33,39,45,57,59,66,69,75,79,82,88,96,100,107,118,129, 132,134,145,162,165,166,173,174,175,179,180,181,182,185, 187,190,197,200,202,177,178,194,195。... |
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