Establishment of seedling classification criteria of Coptis chinensis by dynamic clustering method / 中国中药杂志

<p><b>OBJECTIVE</b>To establish seedling classification criteria of Coptis chinensis.</p><p><b>METHOD</b>The height of plant, leaf number, leaf length, leaf wide, weight of leaf, weight of root were measured, the main measurement indexes of seedlings of C. chinensis were chosen through correlation and regression analysis. The seedling classification criteria were formulated by dynamic clustering analysis.</p><p><b>RESULT</b>The criteria of the 1st-grade seedlings were as follows: leaf number above 8 leaves, height of plant between 12 and 14 cm. The criteria of the 2nd-grade seedlings were as follows: leaf number between 6 and 8 cm, height of plant between 9 and 12 cm. The criteria of the 3rd-grade seedlings were as follows: leaf number between 4 and 6 cm, height of plant above 9 cm.</p><p><b>CONCLUSION</b>The seedling classification criteria of C. chinensis was scientific and feasible, and can be used for the quality control standard of C. chinensis.</p>

Quality classification standard of Dipsacus asperoides seed / 中国中药杂志

<p><b>OBJECTIVE</b>To establish seed quality classification standard of Dipsacus asperoides.</p><p><b>METHOD</b>Through the detection on seed purity, 1 000-grain weight, water content, germination rate of D. asperoides from different areas, and observation on seed external characters, the primary seed quality classification standard of D. asperoides was preliminarily formulated.</p><p><b>RESULTS</b>The first level D. asperoides seed germination rate was over 85%, 1 000-grain weight above 3.94 g, purity above 90.95%, water content lower than 9.08%. The second level D. asperoides seed germination rate was over 64%, 1 000-grain weight was above 3.57 g, purity was over 83.66%, water content was above 10.23%. The third level seed germination rate was above 35%, 1 000-grain weight was above 3.04 g, purity was above 75.51%, water content was lower than 11.37%.</p><p><b>CONCLUSION</b>Germination rate and 1 000-grain weight were the main indexes of quality classification standard, and purity and water content provide the important reference. This quality classification standard of D. asperoides was scientific and feasible, and can be used as the quality control standard of D. asperoides.</p>

Analysis of genetic diversity and genetic relationships of Lonicera macranthoides cultivars / 中国中药杂志

<p><b>OBJECTIVE</b>To study genetic diversity and genetic relationships among Lonicera macranthoides cultivars.</p><p><b>METHOD</b>Five cultivars were estimated by ISSR and SRAP. The data of amplified bands were analyzed by Treeconw software. The system diagram of genetic relationship was built by UPGMA.</p><p><b>RESULT</b>Twenty ISSR primers amplified 186 bands with 103 (54.63%) polymorphic bands and 58 SRAP primer combinations amplified 591 bands with 347(55.46%) polymorphic bands. Genetic distance ranges were 0.058 4-0.230 8 (by ISSRs) and 0.1071-0.2611 (by SRAPs). Both ISSR and SRAP analyses revealed a middle level of genetic diversity in L. macranthoides cultivars. The dendrograms based on SRAP and ISSR markers were not all the same.</p><p><b>CONCLUSION</b>The genetic diversity of L. macranthoides cultivars is middle. ISSR and SRAP markers can be effectively applied to genetic analysis in L. macranthoides cultivars.</p>

Genetic diversity of Scrophularia ningpoensis based on SCoT analysis / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the genetic diversity and relationship of different Scrophularia ningpoensis cultivars.</p><p><b>METHOD</b>Forty-eight germplasmic resources of S. ningpoensis cultivars were analyzed by Start Codon Targeted Polymorphism(SCOT) molecular markers. Genetic distance was calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method.</p><p><b>RESULT</b>A total of 279 bands were detected using 48 primers, among which 214 were polymorphic bands. The average percentage of polymorphic bands was 76.7%. Genetic distance was changed from 0.1507 to 0.4933. Clustering results showed that the genetic relationship of S. ningpoensis cultivars was more complex. There was significant correlation between some germplasm and its geographic origin while geographical distribution of some germplasm was not very obvious, but it was also showed that some of the S. ningpoensis from the same region were in the same group which presented the law of geographical distribution in the tested materials.</p><p><b>CONCLUSION</b>Significant polymorphism and genetic diversity can be observed among S. ningpoensis germplasm resources which provided a wealth of genetic basis for cultivating fine varieties.</p>

Analysis of genetic relationships of Pueraria thomsonii based on SRAP markers / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the interspecies relationships of 18 Pueraria thomsonii cultivars in molecular level.</p><p><b>METHOD</b>Eighteen P. thomsonii cultivars were evaluated by using sequence-related amplified polymorphism (SRAP) markers, with P. lobata and P. peduncularis as contrast species. Systematic relationships were constructed based on the UPGMA method by TREECONW software.</p><p><b>RESULT</b>The results showed that 22 primer pairs produced 338 loci, out of which 216 were polymorphic, the percentage of polymorphic loci was 63.9%. An average of 15.4 loci and 9.8 polymorphic loci were generated by each pair of primers. Genetic distance was analyzed by TREECONW software. Genetic distance of 18 P. thomsonii were changed from 0.004 7 to 0.265 8, with an average of 0.316. Using cluster analysis (UPGMA) based on those polymorphism bands amplified with SRAP primers, the 22 cultivars were classified into three groups, groups 1 with 18 P. thomsonii, group 2 with 3 P. lobata, and group 3 with 1 P. peduncularis. Most of the P. thomsonii from the same region were not in the same group.</p><p><b>CONCLUSION</b>SRAP markers could be a good marker for genetic relationship research in the P. thomsonii.</p>

Establishment of canditate core collection of Artemisia annua from China / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the candidate core collection of Artemisia annua.</p><p><b>METHOD</b>The morphologic traits and the results of SRAP marker were compared respectively.</p><p><b>RESULT</b>The chi2 test and the t test of both indexes for 8 morphological characters and SRAP marker parameters did not reach significant level between the candidate core collection and the primary sample.</p><p><b>CONCLUSIONS</b>In this study, the candidate core collection can stand for original collection excellently.</p>

Analysis of genetic difference among Scrophularia ningpoensis cultivars by SRAP / 中国中药杂志

<p><b>OBJECTIVE</b>The genetic difference among Scrophularia ningpoensis cultivars were analyzed in molecular level.</p><p><b>METHOD</b>Ninety-two individuls of three S. ningpoensis cultivars were employed to be analyzed by the approach of Sequence-related Amplified Polymorphism (SRAP).The parameters were calculated by POPGENE1.31 and the relationship was constructed based on UPGMA method.</p><p><b>RESULT</b>1) A total of 227 bands were scored and 199 bands of them were polymorphic. 2) The result is showed that there is a medium level of genetic diversity among three cultivars. At species level: percentage of polymorphic loci PPB=52.42%, effective number of alleles N(e)=1.2812, Nei's gene diversity H=0.1671 and Shannon's information index H(sp)= 0.2526; At cultivar level: PPB=21.44%, N(e)=1.1216, Nei's gene diversity H=0.0725 and Shannon's information index H(pop)= 0.1083. 3) The Nei's coefficient of genetic differentiation was 0.5625, which was consistent with the Shannon's coefficient of genetic differentiation (0.5713). Most of the genetic variation existed among cultivars. 4) The gene flow (N(m)=0.3889) was less among cultivars, indicating that the degree of genetic differentiation was higher. 5) Genetic similarity coefficient were changed from 0.8082 to 0.9133. By clustering analysis, the classified result of SRAP marker between traditional modal character was almost same.</p><p><b>CONCLUSION</b>The genetic diversity of samples of S. ningpoensis is medium. The genetic difference among cultivar is higher than that within cultivar.</p>

Study on genetic diversity of Codonopsis tangshen by SRAP and ISSR markers / 中国中药杂志

<p><b>OBJECTIVE</b>To study the genetic diversity of Codonopsis tangshen.</p><p><b>METHOD</b>Eighteen germplasmic resources of C. tangshen were analyzed by SRAP and ISSR molecular markers. The systematic diagram of genetic relationship was made by TREECONW software and clustered by UPGMA method.</p><p><b>RESULT</b>Twenty-nine SRAP primer combination amplified 329 bands with 266 (80.85%) polymorphic and 21 ISSR primers amplified 223 bands with 166 (74.44%) polymorphic. The average genetic similarity coefficient was 0.7121 (by SRAPs) and 0.7781 (by ISSRs). Both SRAP and ISSR analyses revealed a high level of genetic diversity in C. tangshen. By cluster analysis, the geographical distribution was not distinctive. The significant positive correlation between SRAPs and ISSRs was observed (r=0.802, P<0.01), although the dendrograms based on SRAP and ISSR markers were not all the same.</p><p><b>CONCLUSION</b>Different germplasms diversity of C. tangshen is high and SRAP and ISSR can be used in genetic diversity study of C. tangshen.</p>

Genetic relationship among populations of cultivated Coptis chinensis revealed by SRAP / 中草药

Objective To reveal the genetic relationship among populations of cultivated Coptis chinensis.Methods Twenty four populations of cultivated C.chinensis from different habitats were employed to be analyzed by the approach of sequence-related amplified polymorphism(SRAP).Systematic relationship was constructed based on the UPGMA method by Treeconw software.Results A total of 276 bands were scored,among which 120 were polymorphic bands.The average percentage of polymorphic bands was 43.48%,indicating that the materials in the test have low genetic diversity.Genetic similarity coefficients were changed from 0.877 0 to 0.951 9.By cluster analysis,the geographical distribution was not very obvious,but it was also showed some of the cultivated C.chinensis from the same region were in the same group.Conclusion Different germplasms diversity of cultivated C.chinensis population is lower and genetic background is more single.

DNA Extraction from Rhizoma Coptidus and optimization of RAPD reaction system / 中草药

Objective To study the genomic DNA extraction from Rhizoma Coptidus and optimization of RAPD reaction system. Methods Different methods, i.e. phenol method, CTAB method, low pH extraction medium with high salt, were used to genomic DNA extract from Rhizoma Coptidus. The DNA samples obtained by the above methods were tested by agarose gel electrophoresis and ultraviolet spectrometer. Results CTAB Method was considered to be an optimal technique. Based on the genomic DNA extracted by CTAB method, a reaction system suitable for Rhizoma Coptidus was established, that is, 25 ?L amplification reactions system containing 1?PCR buffer, 2 mmol/L Mg 2+, 100—150 ?mol/L dNTP, 20 ng primer, 40 ng template DNA, and 1 U Taq DNA polymerase. Conclusion CTAB Method and RAPD reaction system can be used to RAPD analysis in Rhizoma Coptidus.