| In this study, we used 60 clones of Pinus koreansis as research materials from four locations of Korean pine seed orchards in Heilongjiang province. We analyzed genetic variation, genetic parameter estimation indicators and related analysis by studying clone strong character, a real properties nutritional composition, pine nuts, seeds nutrients, fatty acids composition in the pine nut oil and amino acid composition in protein in each seed orchard. We studied the diversity free pollination levels of Korean pine progeny and obtained Korean pine SSR markers by using high-throughput sequencing. Our purpose was studied strong character genetic variation, kind of real character, genetic variation rule of nutrients between different clones. The results could provide important traits for Korean pine breeding. The correlation between indicators would reflect the degree of interaction and provide powerful basis for high quality generation breeding, early selection of Korean pine and shorten the breeding period. We could also evaluate productivity level of clonal seed orchard and estimate genetic parameter correctly. Through multiple selection method, we screened superior quality clones from Heilongjiang province. We also provided construction materials for local clonal breeding and clonal seed orchards.1. The genetic variation of characterThe results of variance analysis indicated that indicators such as seeds character of Korean pine, seeds nutrients, fatty acid in pine nut oil and amino acid composition of protein of existed a significant difference between clones. The variation of seed morphological characters was from 8.40% to 17.81%. The variation coefficient of seed long was the minimum and the variation coefficient of seed weight was the maximum. The content of oil in nutrient composition of seeds was the highest. The variation range of nutrient composition was form 10.79% to 10.79% in seeds. There were 11 kinds of fatty acids in oil of pine nuts, which including 5 kinds of saturated fatty acids and 6 kinds of unsaturated fatty acids. The content of linoleic acid was the highest in fatty acids, which reached the level of 42.02%. The variation amplitude of fatty acid composition was from 2.24% to 66.83%. Seventeen kinds of amino acid composition were detected in seeds protein. The average mass fraction of total amino acids was 40.43%. The average mass fraction of the total essential amino acids was 9.79% which accounted for 24.21% of total amino acids. The variation range of amino acid composition was from 14.70% to 38.88%%.2. The correlation analysis between charactersThe relationship between traits of seed orchard, which including traits phenotypic correlation and genetic correlation. Though discussed the relationship between traits of seed orchard, we found out that there were different degree of correlation between different characters in Korean pine clones. Under different site conditions, some of the traits of phenotypic correlation and genetic relationship were very stable. There was positive correlation between genetic correlation and phenotypic correlation of grains, seeds weight, tare weight and seed length in four different seed orchards. There were significantly positive correlation between seeds weight, tare weight and seed length. There were significantly negative correlation between nutritional components of carbohydrate, fat content, protein content and amylose content. There were weak positive correlation between oil, protein and polysaccharide. Fatty acid composition of Σ SFA C18:1, Σ MUFA and Σ USFA phenotypic correlation and genetic correlation were significantly negative relationship. C18:1 with Σ MUFA and Σ USFA presented a very significant positive correlation. C18:l with C20:1, C18:2, C18:3, C20:2 and Σ PUFA were significant negative correlation relationship. Σ MUFA with USFA and C18:1 were significantly positive correlation. Σ PUFA with C18:2, C18:3, C20:2 and C20:1 were significantly positive correlation. Σ PUFA with C18:1 and Σ MUFA were significantly negative correlation. There were significant positive correlation between acid composition of TAA, EAA, NEAA, Thr, Ser, Val and Phe. There were very significant positive correlation between TAA, EAA, Thr, Ser, Glu, Gly, Leu, Tyr, Phe, Lys, His, Arg and Val. There were significant positive correlation between Arg, Thr, Ser, Glu, Gly, Leu, Tyr, Lys and His. In indirect selection of traits, breeding objectives should be considered and achieved the purpose of comprehensive select optimal finally. After selection and operation, we choosed the kernel, grain and high nutrient of clones.3. The study of clonal selectionWe evaluated the character from the whole area and selected the mean of clone more than one standard deviation of each character for inclusion criteria to estimate the genetic rate of Korean pine solid character of population from 21.88% to 94.81% in clone source. Through character selection, we selected intensity between 1.37 to 1.75, properties of genetic gain from 1.42% to 51.71%, the minimum of length-width ratio in genetic gain at 1.42% and the rest of the character of genetic gain was more than 10%. We estimated that the genetic rate of fatty acid composition in clone population was from 47.64% to 85.19%. Through character selection, we selected intensity between 1.21% and 32.91%, estimated genetic rate of clone population in the amino acid composition of pine nuts was from 42.2% to 98.51% and selected intensity between 1.27 and 1.56, and genetic gain between 9.35% and 70.3%. Through studying the selection of superior clones and asexual reproduction, the results obtained higher genetic gain and explained strong genetic control.By clonal test for each seed orchards, the clonal repetition rates were in middle and high levels. Two kinds of evaluation methods, index selection method and principal component analysis, were used to breeding clones. According to 20% clone year-old open pollination progenies, we combined with two methods to select three clones as good clones in Hegang (HG8, HG21 and HG14), Linkou (LK20, LK19 and LK18), Tieli (TL1104, TL113land TL1383) and Weihe (WH071, WH117 and WH066) seed orchards, respectively. We could achieve better effect by selecting clones. The mean index was higher than total average of clone population in the kernel, grain, seeds weight, fat, protein, Σ PUFA, EAA and TAA.4. Analysis of genetic structureWe developed 16 pairs of SSR primers by high-throughput sequencing of Korean pine. Preliminary, we tested fluorescent markers of by using 6 pairs of primer form free pollination progenies of Korean pine seed orchard family. The results showed that the generation of Korean pine seed garden had moderate polymorphism level. The average of polymorphism information (PIC) was 0.325. The levels of diversity were detected in four generation seed orchards. The level of diversity (I= 0.7488) and observed heterozygosity (Ho= 0.4848) were the highest in Weihe seed orchard.Through studying phenotypic differentiation and genetic differentiation in Korean pine, we found out that the mean coefficient of phenotypic differentiation was 39.69% in each character index of Korean pine. Gst was 0.0387, which estimated from differentiation coefficient of Nei index. The results of phenotypic differentiation and genetic differentiation showed that genetic differentiation was mainly existed between clones in Korean pine seed orchards. |
PDF Full Text Request