| Ziziphus jujuba 'Lingbaodazao' which is a fine local variety with big fruit, high sugar, nutrient-rich, and has a long history cultivated, in the long-term cultivation process, because of climate, soil and other natural conditions and the impact of human activities, their morphological characteristics, biological characteristics and quality features of the natural are significant variation of Lingbao jujuba production standardization and commercialization of adverse effects, but also for the breeding of improved strains from which laid a favorable foundation. In this paper, the traditional morphological markers combined with technology is mature and stable economy with high levels of applicable and polymorphism of ISSR markers, the genetic diversity Lingbao jujuba, the relationship between genetic structure and genetic studies, collecting some fine individual and carrying out grafting experiments, the Lingbao jujuba breeding, germplasm conservation provide a theoretical basis. The main results are as follows:1. Ziziphus jujuba 'Lingbaodazao' material on 70 of 28 phenotypic analysis showed that the coefficient of variation: 14 Ziziphus jujuba 'Lingbaodazao' plant traits, the average coefficient of variation was 17.36%, the average variance of 9.3267; 8 traits of the average fruit coefficient of variation was 25.29%, the average variance is 15.1270; 6 characters inclusions, the average coefficient of variation was 11.39%, the average variance is 8.1293. Shows that Ziziphus jujuba 'Lingbaodazao' fruit phenotypic variation of the total variance in the highest degree of plant variation center, while the inclusion of the most stable, the coefficient of variation is also minimal.2. Phenotypic characteristics of cluster analysis showed that: 70 materials were divided into three groups, the first group for large fruit, as a typical representative of the type, and the second group of small fruit type and long-shaped fruit, these are rare species. The third group only one, different from many other plant morphology, might be a variety in the population or homonym. With the 10 main characters Ziziphus jujuba 'Lingbaodazao'correlation analysis showed that: fruit shaped largely by the impact of fruit diameter; fruit weight and content of inclusions close relationship, inclusion relationship between the characters is not obvious.3. The improved SDS method is the best one with fewer steps, easy to handle and stable. The average OD260/280 1.77; the average yield was 93.7μg / gThe extraction is suitable acting as template DNA for ISSR-PCR amplification.4. The optimal ISSR-PCR reaction system was determined.Each 20uL reaction system should contain 80ng template DNA , 0.7mmol/L primer , 10.0μL 2×Taq PCR MasterMix.The amplification program as follows:predenature at 94℃for 5min,followed by denature at 94℃for 45s,annealing at 54℃(depend on primer ) for 1 min,extend at 72℃for 90s,35 cycles,and then extend at 72℃for 8min, final store at 4℃for ever.5. 12 stable polymorphisms primers were selected from 100 ISSR primes, 70 Ziziphus jujuba 'Lingbaodazao' on genomic DNA was amplified to a total out of 105 sites per primer locus 8.7 a, 91 polymorphic sites, the percentage of polymorphic loci was 86.67%, the number of alleles na = 1.8667, the effective number of alleles ne = 1.5491, gene diversity h = 0.3183, Shannon information index I = 0.4725 shows that between 70 Ziziphus jujuba 'Lingbaodazao' material there is a high genetic variation.6. Population genetic structure analysis shows that: Ziziphus jujuba 'Lingbaodazao' group gene diversity Ht was 0.3193, gene diversity within populations Hs was 0.2828, gene diversity among populations Dst was 0.0365, coefficient of gene differentiation Gst was 0.1144, that indicating only 11.44% of the variation comes from the population between the main genetic variation among individuals within populations and gene flow Nm to 3.8717> 1, indicating that gene flow among populations is large,therefore,the differentiation between populations is relatively small.7. Based on ISSR data of cluster analysis with cluster analysis based on phenotypic results were inconsistent, but basically indentical; principal coordinate analysis revealed Ziziphus jujuba 'Lingbaodazao' genetic relationship between the various materials for futher; on the genetic distance and the geographical relationship of the correlation analysis showed that genetic distance and geographic distance has a weak correlation.8. Comprehensive phenotypic and molecular characteristics of an integrated morphological and molecular cluster, from 70 to select 6 fine individuals, with grafting methods for remote testing and species comparison determination of genetic stability, the results need to be observed further. |
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