| Common bean (Phaseolus vulgaris L.) has the largest cultivation area among edible beansin the world, and is also the very important edible vegetables in China, which is rich in nutrition,and contains plenty of quality starch and protein. The common bean is cultivated in most partsof China. Northeast of China is one of the main common bean producing areas with abundantspecies. This study made the goal to clarify bean germplasm diversity and lay the initialfoundation of collection, using, identification, genetic map construction and classification ofcommon bean germplasm by studied the agronomic traits and ISSR molecular markers of36varieties of common beans certified recent years and main cultivated in Jilin province. Theresults of this study are obtained as follows:1. The diversity anylysis by morphological markers on the seeds, fruits and plants of the36tested varieties showed that the genetic diversity was very rich. The variation coefficient ofmorphological indexes changed in the range of8.33%-31.35%. The variation degree of fruitswas higher than seeds and plants. The variation coefficient of physiological indexes changed inthe range of0.99%-75.18%. The difference of Vc content between varieties was the largest, onthe contrary, the difference of moisture content was the smallest. The molecular markers resultsshowed that the polymorphism rate of amplified polymorphic was87.36%, but the rangeabilityof similarity coefficient between materials was small.2. The correlation analysis on the relevant traits of tested materials showed that the fruitlength of common bean was highly significantly positive correlated with seed lenth, and highlysignificantly negative correlated with seed thickness. There was a highly significantly positivecorrelation between fruit width and seed width. The fruit beak length was highly significantlypositive correlated with seed length and seed width. The negative correlation between sarcocarpthickness and seed width was highly significant. The leaf width was significantly negativecorrelated with fruit width and fruit beak length. The fruit thickness was significantly positivecorrelated with plant height. The node of the first flower had no significantly correlation withfruit characters.The soluble sugar content was significantly negative correlated with fruit width and fruitweight. There was a significantly negative correlation between soluble protein content and fruitwidth. The positive correlation between chlorophyll content and fruit color was significant, andthe positive correlation between chlorophyll content and fruit width was highly significant. The nitrate nitrogen content was significantly positive correlated with fruit beak length, andsignificantly negative correlated with fruit thickness.3. The cluster results of tested materials were not exactly the same between the cluster onthe basis of morphological index, quality index and molecular marker methods. But the samewas the No.32, No.33and No.34varieties were clustered in one group, which showed that thephylogenetic relationships of the three varieties were closer. Through the comprehensiveanalysis of the morphological index and the molecular markers indicated that the varieties withcloser phylogenetic relationships were: No.29and31varieties, No.2,3,5,6,7,8and17varieties, No.10-16varieties, No.21-24varieties, No.29and31varieties. The results showedthat the varieties from the same source were tended to be clustered in one group. By thecomprehensive analysis of the quality index and molecular markers indicated that the varietieswith closer phylogenetic relationships were: No.2,7and14varieties, No.1,3,4,5,6,8,12and13varieties, No.9,10,15and17varieties, No.18,19,21,22,23and24varieties, No.25-30varieties. The results indicated that the varieties of different sources also had closephylogenetic relationships.4. The optimum ISSR-PCR reaction system for common bean was established: with25μLtotal reaction volume contains45ng DNA template,1.5U Taq DNA polymerase,2mmol/L Mg2+,0.5μmol/L ISSR primer and0.15mmol/L dNTP, other components were sterile double distilledwater, and the suitable cycles was45cycles.The ISSR amplification procedure of common bean was: after pre denaturing at94℃for5min, then followed by40cycles of procedures: denaturing at94℃for40s, annealing at54℃for45s, extending at72℃for1min20s; and finally extending in72℃for8min, then terminationreaction and hold at4℃.20primers were screened out from the100ISSR primers for common bean: primer812,primer818, primer823, primer824, primer834, primer835, primer840, primer842, primer846, primer856, primer857, primer860, primer861, primer867, primer880, primer881,primer888, primer895, primer899, primer900.3specific primers were screened out: thespecific makers of cultivar20was primer840, the specific makers of cultivar27was primer861,and the specific makers of cultivar31was primer846. The specific primers above could quicklyidentify cultivar20,27and31from all the36common bean varieties. |
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