The Study On Germplast Resources Diversity Of Scrophularian Ningpoensis Hemsl

Scrophularian ningpoensis Hemsl. was a kind of crucial medical herb. Yet it is a long time that the harvest was blindfold and unplanned for its wild resources, which resulted in breads being farraginous for cultivars and germplasm resources were destroyed baldly. So it was urgent for studying and protecting the germplasm resources.Six populations of Scrophularian ningpoensis Hemsl. from five different regions were selected in the research. First the morphological characters (including leaves length, leaves width, ratio of L/W, plant height and diameter of root) were mensurated, then the DNA fingerprintings were constructed and their genetic distance were calculated, at last we mensurated the medical content in their former regions and Zhenping, Shaanxi Province for Scrophularian ningpoensis Hemsl., and combining the morphological characters we took cluster analysis for the seven indexs, the results are as follow:1. The cluster chart was drawed based the morphological characters and medical content, and six populations were divided into four branches: A2 (Badong Xuanshen) and A4 (Nanchuan Xuanshen), A3 (Anguo Xuanshen) and A5 (ZhenpingXuanshen) were the same branches respectively, and A1 (Dongyang Xuanshen) was a branch, A6 (Wild Xuanshen) became a branch by itself. From the cluster chart we can see that the former regions of A2 and A4 were not far from each other (Hubei and Chongqing), so their cluster distance was the nearest. What is more, A3 and A5 were also the nearest in cluster distance, their ratio of L/W, plant height and diameter of root were similar, and their former regions were near (in Henei and Shaanxi) compared with others. The cluster distance of A1 was very far compared with A2-A5, and its leaves length, plant height were the least, the content of Arpagoside was low, yet the content of Cinnamic Acid was the highest, the phenomena may be caused by the special environment of Zhejiang. A6 was discovered in Zhenping, and it was special from morphological characters to medical content, so its cluster distance must be the furthest.2. We established and optimized the ISSR-PCR reaction system and amplication procedure. And the amplifications were carried out best in 10×buffer, 0.3mmol/L dNTPs, 0.5-1.0U of Taq DNA polymerase, 2.0mmol/L MgCl₂, 0.4μmol/L primer, and 150ng of genomic DNA per 20μL reaction system. Initial denaturation was for 5 min at 94℃, followed by 35 cycles of 30 s at 94℃, 45 s at 50-61℃(following the different primers), 1 min at 72℃, and a final 7-min extension at 72℃.3. The DNA fingerprintings were established, which could distinguish the different populations of Scrophularian ningpoensis Hemsl. based on the screened seven primers (807,811,818,825,828,836 and 890), at the same time the genetic cluster chart was drawed according to the different genetic distance. For the cluster chart, six populations were divided into five branches, the genetic distance of A4 and A5 were the least, so their relationships were the nearest, then the next were A1, A3, and A2; the genetic distance of A6 (0.45655) was furthest compared with others, so its difference was most obvious.4. The content of Arpagoside and Cinnamic Acid were measured. The change scale was 0.063%-0.945% and average value was 0.301% for Arpagoside content, the highest was 14 multiple compared with the lowest; and for Cinnamic Acid the change scale was 0.034%-0.132% and average value was 0.064%, the highest was 2.88 multiple compared with the lowest. At the same time measured the change after A1-A4 were planted in Zhenping, the result showed that the average Arpagoside content increased by 127.02% and the average Cinnamic Acid content decreased by 18.28%. Then we found that the environment of Zhengping is very suitable for Scrophularian ningpoensis Hemsl. planting, so we could take Zhengping as GAP planting base.