Different development phase of transcription proteomics and metabolomics of flower of Lonicera macranthoides / 中国中药杂志

In order to study the regulation mechanism of secondary metabolites biosynthesis in Lonicera macranthoides, the key genes involved in the regulation of biosynthesis and the mechanism of differential metabolites were explored. In this study, high-throughput sequencing technology was used for transcriptome sequencing of L. macranthoides at different development stages. By using Liquid chromatography-tandem mass spectrometry(LC-MS/MS) technology, the laws of qualitative, quantitative and synthetic accumulation of its metabolites were studied, and the key enzyme genes for the biosynthesis of phenolic acid and flavonoids were screened out according to the differentially expressed genes. A total of 111 differentially accumulate metabolites(DAM) and 6 653 differentially expressed genes(DGE) were obtained by metabonomics and transcriptomics analysis. The metabolites and key enzyme genes in the Erqing(KE) were significantly different from those in the Dabai(KD) and Yinhua(KY) stages. In the phenylalanine biosynthesis pathway, the ion abundance of chlorogenic acid, naringin, quercetin, rutin, coniferol and other metabolites decreased with the development of flowers, while the ion abundance of ferulic acid, coumarin and syringoside increased with the development of flowers. Key enzyme genes such as CHS, HCT, CCR, FLS and COMT positively regulate the downstream metabolites, while PAL, C4H and 4CL negatively regulate the downstream metabolites. This study provides candidate genes and theoretical basis for the further exploration of key enzymes in the biosynthesis of secondary metabolites and for the regulation of the accumulation of secondary metabolites in L. macranthoides by molecular biological methods.

Transcriptome analysis of Platycodon grandiflorum at different growth years and discovery of key genes for platycodin biosynthesis / 中国中药杂志

Platycodon grandiflorum is a medicinal and edible medicinal material. Our study is aimed to explore the differences in the gene expression of P. grandiflorum in different growth years, and the expression rules of key genes in the biosynthesis of the main active substances of P. grandiflorum. Illumina Hiseq 4000 sequencing platform was used to sequence the transcriptome of P. grandiflorum in different years. Then, 59 654 unigenes were obtained through filtering, assembly, splicing and bioinformatics analysis of the sequencing data, of which 1 671 unigenes were differentially expressed between at least two samples. The results of cluster analysis showed that there was a great difference in the gene expression of P. grandiflorum from one-year-old to two/three-year-old. There were 1 128 different genes between one-and three-year old P. grandiflorum, and only 57 different genes between two-and three-year-old P. grandiflorum. KEGG enrichment results showed that the differential genes of P. grandiflorum in different years were mainly concentra-ted in the biosynthesis of sesquiterpenes and triterpenes, and the biosynthesis of terpenoid skeletons. In the triterpenoid biosynthesis-related pathways, a total of 15 unigenes were identified, involving 5 enzymes. The expression levels of ACAT, HMGR, FDFT1, SQLE decreased with the increase of the growth year of P. grandiflorum. The expression of HMGS was the highest in the one-year-old P. grandiflorum, followed by the three-year-old sample. This study provides useful data for the development of P. grandiflorum, and also provides a basis for the study of related genes in the biosynthetic pathway of platycodin.

Variation in physicochemical properties and bacterial community structure in rhizosphere soil of Coptis chinensis tow cropping modes / 中国中药杂志

The natural forest and artificial shed are the main cropping modes of Coptis chinensis. This study is aimed to reveal the rhizosphere soil bacterial community structure difference between under tow C. chinensis cropping modes-natural forest and artificial shed, and to assist us to completely understand soil quality condition,and provide theoretical guidance for soil improvement and C. chinensis planting. The rhizosphere soil samples of 1-5-year-old C. chinensis under tow cropping modes-natural forest and artificial shed were collected. Illumina high-throughput sequencing technology was used to analyze the alpha diversity, community composition, community structure of soil bacteria under the tow cropping modes,and the effects of soil nutriment indices on soil bacterial community structure. Through the analysis of species number, Shannon, Chao1 index and ACE index of bacterial community, it was found that the bacterial diversity of 1-year-old C. chinensis soil under natural forest cropping mode was significantly lower than that under artificial shed cropping mode, and the diversity of bacterial communities in soil of 2-5-years old C. chinensis were not significant different between two cropping modes. A total of 53 phyla,60 classes,140 orders and 266 families were detected in the rhizosphere soil of C. chinensis under the cropping modes of natural forest, respectively. The rhizosphere soil of C. chinensis under the cropping modes of artificial shed included 54 phyla,65 classes,140 orders and 264 families, respectively. Under the two cropping modes, the top 10 dominant species of bacterial community abundance are the same, they are Proteobacteria, Acidobacteria, Actinobacteria,Bacteroidetes, Planctomycetes, Chloroflexi, Verrucomicrobia, Gemmatimonadetes, Firmicutes and Cyanobacteria, but there are differences in the abundance sequence. The top 10 dominant species of bacterial community abundance accounted for 74.36% to 74.30% of the total bacteria, and 3.15% to 3.92% of the bacteria are unclassified. The results of Metastat analysis showed that the abundance of Gemmatimonadetes in the rhizosphere soil of C. chinensis under the cropping modes the artificial shed was significantly higher than that under the natural forest cropping mode(P<0.05). MRPP analysis of community structure differences showed that under tow cropping modes, there were significant differences in the bacterial community structure of 1-4-year-old soil bacteria, among which the difference between 1-year-old soil samples was the largest. With the increase of cropping years, the difference gradually decreases, and there is no significant difference in the bacterial community structure between 5-year-old soil samples. RDA analysis and correlation analysis of bacterial community structure and soil physical and chemical properties showed that the order of environmental factors on the rhizosphere soil bacteria of Coptis chinensis was: pH>available P> total P> total K>bulk density>total N>available N>organic matter. The results are helpful to understand the soil health of C. chinensis and provide scientific basis and theoretical guidance for soil improvement and C. chinensis planting.

Comparative transcriptomics analysis of Citrus medica var. sarcodactylis at different developmental stages / 中国中药杂志

In order to study the transcriptional differences of Citrus medica var. sarcodactylis at different developmental stages, we explored the genes regulating the biosynthesis of the effective components. In this study, Illumina Hiseq 4 000 high-throughput sequencing technology was used to sequence the transcriptome of C. medica var. sarcodactylis at different developmental stages, 121 235 unigenes were obtained with an average length of 2 434 bp, 3 379 different genes were obtained using DESeq screening, which mainly connected to biological processes such as signal transmission, biological regulation, and metabolic processes, and enriched in metabolic pathways such as starch, sucrose metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis. Further dynamic comparison of biosynthesis related genes of active ingredients: the expression levels of PAL, CHI, CYP75B1, ZDS, 4CL and FLS gradually increased as the fruit turned from green to yellow; the expressions of COMT, F3H and CYP73A increased at first and then decreased; CCR, HCT and HRP were down-regulated whereas up-regulated. This study provides references for further excavation of key genes in the biosynthesis of active components, as well as biopathway analysis of active components for C. medica var. sarcodactylis.

Variation in fungal community structures in rhizosphere soil of Coptis chinensis with cropping mode under natural forest and artificial shed / 中国中药杂志

This study is aimed to reveal the rhizosphere soil fungal community structure difference of Coptis chinensis cropping between natural forest and artificial shed modes, and provide theoretical guidance for soil improvement and C. chinensis planting. The rhizosphere soil samples of 1-5-year-old C. chinensis under natural forest and artificial shed modes were collected. Illumina high-throughput sequencing technology was used to analyze the community structure and diversity of soil fungi under the tow cropping modes,and the effects of soil nutriment indices on soil fungal community structure. The results suggested that the abundance and diversity of fungal communities in soil of 2-5-year-old C. chinensis were not significant different in both two cropping modes, but it was significantly higher than that in the 1-year-old C. chinensis. Comparing soil samples from the same year-old C. chinensis under the two cropping modes, it was found that there was no significant difference in the abundance and diversity of fungal communities. The fungal community of the rhizosphere soil was different in composition and abundance between tow cropping modes, and between different planting years. The 17 phyla,59 classes and 155 orders,and 17 phyla,59 classes and 157 orders were detected in the rhizosphere soil of C. chinensis under the cropping modes of natural forest and the artificial shed, respectively. Ascomycota, Basidiomycota and Mortierellomycota were dominant phyla in rhizosphere soil, and the average abundance of the 3 phyla accounted for 74.36% and 74.30% of the total fungi. The results of analysis of similarities showed that there were significant differences in the fungal community structure of 1-year-old and 2-year-old C. chinensis soil fungi, and there was no significant difference in the community structure of 3-5-year-old samples. Under the natural forest cropping mode, there were significant differences among the samples of different years. Under the artificial shed cropping, there were significant differences in fungal community structure between 1-year-old and 3-5-year-old C. chinensis soil, and between 2-year-old and 3-5-year-old C. chinensis soil. The results of canonical correlation analysis showed that soil pH and soil organic matter content were the main factors affecting the soil fungal community structure. Soil organic matter content was positively correlated with Basidiomycota and Cryptomycota, pH was negatively correlated with Basidiomycota and C. ryptomycota. The planting of C. chinensis has promoted the diversity and abundance of rhizosphere fungal community significantly. For the same year-old C. chinensis soil, abundance of fungal community was no significant difference between two cropping modes. There are significant differences in the rhizosphere soil fungal community structure between tow cropping modes in the first two years of planting. Through the interaction between the rhizosphere and the soil and the continuous selection of the rhizosphere to the fungal community, the fungal community structure tended to be the same between the two cropping modes in rhizosphere soil of 3-5-year old C. chinensis. The soil pH and orga-nic matter content were the main factors affecting the change of fungal community structure.

Analysis on genetic relationship of medicinal species of Dipsacus marked by ISSR, SCoT, and SRAP / 中草药

Objective To detect the genetic relationship among nine medicinal species of Dipsacus in China. Methods The polymorphic bands were detected by three labeling methods (ISSR, SCoT, and SRAP). The genetic distances were calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method. Results The percentage of polymorphism by ISSR, SCoT and SRAP markers showed little difference with value at 90.4%, 88.5%, and 88.2%, respectively, which indicated that the genetic polymorphism of the tested materials was very rich. The genetic distance between Dipsacus chinensis and D. japonicus was the largest, which indicated their genetic relationship was the most distant. Nine medicinal species of Dipsacus were all divided into three groups by three markers, that D. chinensi and D. lijiangensis, D. asperoides and D. japonicas were respectively clustered into one group, D. asperoides var. omiensis was alone clustered, the other four species were clustered into one group. The clustering results labeled by ISSR and SRAP were basically the same with a slight difference between D. daliensis and D. asperoides. Conclusion The clustering results by there markers was between well consistent with the classical classification, which confirmed that molecular markers can be used as one of the effective methods to reveal the genetic relationship among medicinal species of Dipsacus.

Research for dynamic changes of growth and alkaloids of Coptis chinensis / 中国中药杂志

With Coptis chinensis in high-yielding soil as the object,the growth regularity of plant and dynamic change of alkaloid content was studied. The plant growth model of C. chinensis was constructed. The plant height equation was =3.030 9+0.732 6-0.009 6²,the number of leaves equation was =111.882 6-2 234.881 7/+15 218.960 8/²-31 740.960 8/³,the leaf area equation was =-217.136 1+30.552 2-0.359 0²,the roots talk biomass equation was =-2.748 8+0.210 3+0.006 4²,the number of rootstalk equation was =-1.246 0+0.192 6+0.000 8²,the fibrous root biomass equation was =-4.973 5+0.589 4 -0.002 6². The results indicated that the number of leaves and leaf area were increasing continuously after seedling transplanting,the leaf area of 3-year-old C. chinensis reached a maximum value of 425.83 cm²/plant,after declining.The number of leave of 5-year-old C. chinensis reached a maximum value of 70.91. With the increasing of years of growth, the number of rootstalk and rootstalk biomass of C. chinensis was increasing continuously. The biomass growth of 3-year-old and 4-year-old rootstalk was the fastest in the whole development stage of C. chinensis,the annual increase of more than 300%. The change curve of rootstalk number, rootstalk biomass and fibrous root biomass in the whole growth stage was a s-type.The dry matter partition of leafwas the highest in 1-year-old C. chinensis, and then gradually decreased,the change trend of dry matter partition of rootstalk was just the opposite, the dry matter partition of fibrous root increases with the increase of the growing year, reaching the maximum value in 3-year-old, then gradually lower trend. The root-shootratio of 1-year-old C. chinensis was the smallest, then gradually increases, the growth center gradually shifted to the roots from stems and leaves, The weight of underground part of 3-year-old C. chinensis exceeded the aboveground part, the 5-year-old C. chinensis root-shoot ratio reached the maximum value of 1.91:1.With the increasing of years of growth, the contents of coptisine, berberine, epiberberine and palmatine in rootstalk was increasing continuously. The jatrorrhizine content in 2-year-old C. chinensis was significantly lower than that in other years, the content was no significant change after that. The columbamine content reached a maximum value in 3-year-old C. chinensis,then the decreased gradually. The content of magnoflorine gradually increased and reached maximum value in 5-year-old C. chinensis.

Transcriptome analysis reveals candidate genes involved in flavonols biosyhthesis in Sophora japonica / 中国中药杂志

In order to study the pathways of biosynthesis of flavonoids in Sophora japonica, 113 797 unigenes were obtained by Trinity software, with an average length of 803 bp, of which 72 752 unigenes were obtained from the database by high-throughput sequencing, and a total of 38 891 SSR loci were searched. Through the metabolic pathway analysis, we found that there were 135 unigenes involved in the biosynthesis of flavonoids and 959 unigene involved in other secondary metabolic pathways. Further analysis of genes involved in rutin biosynthesis revealed that 24 were associated with CHS, 52 were associated with FLS, and 11 were associated with UFGT. The obtained data of S. japonica transcriptome lays the foundation for studying the pathways of biosynthesis of flavonoids in S. japonica and provides theoretical basis for the formation of the quality of S. japonica.

Analysis on genetic diversity and genetic relationship of medicinal species in Coptis by SCoT / 中国中药杂志

The genetic diversity and genetic relationship among four medicinal species of Coptis were detected by the approach of sequence-related amplified polymorphism (SCoT). The associated genetic parameters were calculated by POPGENE1.31. The systematic diagram of genetic relationship were clustered by TREECONW. The results showed that a total of 434 bands were produced by using 28 primers, of which 430 were polymorphic loci. There was a high level of genetic diversity among species (PPB=99.1%,Na=1.990 6,Ne=1.329 3,H=0.212 2,I=0.337 8). However, genetic diversity was lower within species, the average of genetic parameters wasPPB=16.8%,Na=1.168 2, Ne=1.073 0,H=0.043 7,I=0.067 7. The Nei's genetic differentiation coefficient was 0.794 0, that indicated that most of the genetic variation existed among species. By clustering analysis, different individuals gathered in the same group. The results confirmed that SCoT marker can be used as one of the effective methods to reveal the genetic diversity and relationship among medicinal species of Coptis.

Identification and phylogenetic analysis of endophytic fungi isolated from Scrophularia ningpoensis / 中国中药杂志

The endophytic fungi from root, main stem, branch and leaf of Scrophularia ningpoensis were isolated and identified from Wulong and Chongqing, and the population diversity analysis and phylogenetic analysis were followed. The result indicated that, as to population diversity index, S. ningpoensis from Wulong： leaf>main stem=branch>root, branch from Chongqing>branch from Wulong. Fifty-eight endophytic fungi were obtained, most of which were the pathogens of the plant. Colletotrichum was the prevailing genus, of which C. gloeosporioides and C. boninense were the prevailing strains. Leaf and seedlings might be the main path of infection. Endophytic fungi and pathogen might convert to each other, influenced by such factors as environment, genotype et al.

Transcriptome characterization for Scrophularia ningpoensis based on high-throughput sequencing technology and related genes for synthesis of terpenoid compounds / 中国中药杂志

To investigate the profile of gene function and search for SSR, a new technology of high-throughput Solexa/Illumina sequencing was used to generate the root transcriptome of Scrophularia ningpoensis, and 65 602 036 raw reads were obtained. Based on the bioinformatics analysis and Trinity, 73 983 unigenes were obtained with an average length of 823 bp. The comparison of sequence homology in database showed that 56 389 unigenes had different degrees of homology. A total of 520 metabolic pathways related genes and 191 relDODO transcription factors were identified by the Swiss-Prot, GO, KEGG and COG.The 11 659 SSRs were found by MISA and the highest frequency was AG/CT. In this study, we obtained numerous SSRs to provide references for the study of functional gene cloning and genetic diversity of S. ningpoensis. The key genes involved in the secondary metabolism are the basis for the study of biosynthesis and regulatory mechanism of the secondary metabolites.

Antifungal effects of three medicinal crops on Phytophthora nicotianae / 中国中药杂志

Tobacco black shank is one of the most harmful soil-borne diseases infected by Phytophthora parasitica. In order to probe the control method to this disease, in this study, the mycelial growth rate method was employed to investigate the antifungal effects of extracts from stem-leaf and root, root exudates, and their combination of Scrophularia ningpoensis, Chuanmingshen violaceum and Pinellia ternata The results showed that: ①Stem-leaf and root extracts of S. ningpoensis, C. violaceum and P. ternata exhibited different antifungal activities, and the inhibition increased with the increase of extract concentration. The antifungal effect of S. ningpoensis extracts at 0.5 g•mL⁻¹ was the strongest than other medicinal plants, the inhibition rate of steam-leaf and root extracts reached 74.88%, 69.27%, respectively. The inhibitory effect of C. violaceum and P. ternata was relatively lower, however, there is a significant gain effect after combination of steam-leaf and root extracts of C. violaceum. ②The root exudates of S. ningpoensis, C. violaceum and P. ternata showed fungistasis to Phytophthora nicotianae, and fungistasis was enhanced with the increase of root exudate concentration. The antifungal effect in the order of C. violaceum > S. ningpoensis > P. ternata. ③The antifungal activity of combination of extract and root exudate from S. ningpoensis was similar with the effect of C. violaceum, they were both stronger than P. ternata, and the antifungal activity for three combination were located between the antifungal activity of their extracts and root exudates. S. ningpoensis and C. violaceum can be potentially applied to prevent and control the tobacco black shank.

Study on associated plants and spatial distribution pattern of Dipsacus asperoides populations / 中国中药杂志

In order to understand the associated species and the population distribution pattern, the investigation of 32 sample plotsfrom the main natural distribution area in Dipsacus asperoides community was carried by quadrat method .The results showed that there were 156 species, which belong to 131 genera and 60 families. There were more species in the two dominant families, Asteraceae and Rosaceae. There were many types of associated, but most appeared at a low frequency. The vegetation type were mostly herbaceous and shrub species, which accounted for 77.6% of the total species. The value of t was greater than t0.05 by methods of variance/mean, showed the difference was significant and the distribution pattern of D. asperoides were cluster distribution. The determination results of seven aggregation intensity index also showed that D. asperoides population accorded with the characteristics of cluster distribution（C>1，K>0，Ca>0，m*>1，m*/m>1，I>0，GI>0）.

Progress on molecular biology of Isaria farinosa, pathogen of host of Ophiocordyceps sinensis during the artificial culture / 中国中药杂志

Isaria farinosa is the pathogen of the host of Ophiocordyceps sinensis. The present research has analyzed the progress on the molecular biology according to the bibliometrics, the sequences (including the gene sequences) of I. farinosa in the NCBI. The results indicated that different country had published different number of the papers, and had landed different kinds and different number of the sequences (including the gene sequences). China had published the most number of the papers, and had landed the most number of the sequences (including the gene sequences). America had landed the most numbers of the function genes. The main content about the pathogen study was focus on the biological controlling. The main content about the molecular study concentrated on the phylogenies classification. In recent years some protease genes and chitinase genes had been researched. With the increase of the effect on the healthy of O. sinensis, and the whole sequence and more and more pharmacological activities of I. farinosa being made known to the public, the study on the molecular biology of the I. farinosa would be deeper and wider.

Quality classification standard of Scrophularia ningpoensis seedlings / 中国中药杂志

The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.

Analysis on genetic diversity and genetic relationship of medicinal species in Dipsacus from China by SRAP / 中国中药杂志

The author detected the genetic diversity and genetic relationship within and among eight medicinal species of Dipsacus by the approach of sequence-related amplified polymorphism (SRAP). The associated genetic parameters were calculated by POPGENE 1.31. The Genetic distance was calculated by TREECONW and the systematic diagrams of genetic relationship were clustered by UPG-MA. The results showed that, using 26 primers, a total of 558 bands were produced, of which 539 were polymorphic loci. There was a high level of genetic diversity among species (PPB = 96.59%, Na = 1.9659, Na = 1.3375, H = 0.2143, I = 0.3423). However, genetic diversity was lower within species, the average of genetic parameters was PPB = 6.97%, Na = 1.0697, Na = 1.0311, H = 0.0187, I = 0.0291. The Nei's genetic differentiation coefficient was 0.9126, indicated that most of the genetic variation existed among species. By clustering analysis, different individuals gathered in the same group and the classified result of SRAP marker between traditional modal characters was almost same. The results confirmed that SRAP marker can be used as one of the effective methods to reveal the genetic diversity and relationship among medicinal species of Dipsacus.

Analysis on genetic diversity among wild Dipsacus asperoides by SCoT / 中国中药杂志

To investigate the genetic diversity among wild Dipsacus asperoides in China, 66 germplasmic resources of D. asperoides 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. The results showed that the totals of 181 bands were detected using 20 primers , among which 109 were polymorphic bands. The average percentage of polymorphic bands was 60.13%. Genetic distance changed from 0.030 6 to 0.181 4. The clustering results showed that there was no significant correlation between the classification of the wild D. asperoides and their geographical origin. The relatively high genetic diversity of D. asperoides provides the basis for breeding new varieties.

Genetic structure and genetic diversity of Artemisia annua varieties (strains) populations based on SCoT markers / 中国中药杂志

To reveal the genetic diversity and genetic structure in Artemisia annua varieties (strains) populations, we detected the genetic polymorphism within and among eight varieties (strains) populations (192 individuals) by the approach of Start Codon Targeted Polymorphism (SCoT). The associated genetic parameters were calculated by POPGENE1.31 and the relationship was constructed based on UPGMA method. The results showed that, using 20 screened primers, a total of 145 bands were produced, of which 122 were polymorphic loci. At species level, there was a high level of genetic diversity among eight varieties (strains) populations (PPB = 84.1% ,H = 0.217 3 and H(sp) = 0.341 9). However, at the variety (strains) population level, genetic diversity was lower, the average of genetic parameters was PPB = 41.9%, H = 0.121 5, H(pop) = 0.186 8. The Nei's genetic differentiation coefficient was 0.441 0, indicate that most of the genetic variation in this species existed within the variety populations. The gene flow (N(m) = 0.633 9) was less among populations, indicating that the degree of genetic differentiation was higher. Genetic similarity coefficient were changed from 0.755 1 to 0.985 7. By clustering analysis, eight varieties (strains) were clustered into two major categories and it was also showed the same or similar genetic background varieties (strains) have a tendency to gather in the same group. Results suggest that, in variety breeding, breeders should strengthen the exchange of bred germplasm and increase mutual penetration of excellent genes, which would broaden the genetic base of A. annua.

Growth and developmental rhythm of Scrophularia ningpoensisin southwest middle mountain area of China / 中国中药杂志

Plant samples were collected and investigated periodically. According to the growth of different parts and the characteristics of dry substance accumulation of Scrophularia ningpoensis, the development of S. ningpoensis could be divided into four stages: seeding stage, stem and leaf growth stage, expanding period of root tubers, and dry substance accumulation stage of root tuber. Leaf numbers of S. ningpoensis grew gradually from one at first to 370 at the final stage, main stem leaf were 50 pieces. Leaf size increasesed with the fastest growth at the stem and leaf growth stage, average daily increase amount was 225 cm2. By the middle of August, leaf size reached to 16,270 cm2. Leaf area indexrose sharply in the seeding stage, and remained above 8 among stem and leaf growth stage and expanding period of root tubers, and rapidly reduced to zero in the stage of dry substance accumulation of root tuber. Leaf area ratio has a tendency of obvious dropping. The net assimilation rate had a small change ranges, two small peak were seeding stage and dry substance accumulation of root tuber. The value of specific leaf area was higher in seeding stage, and in the earlier stage of dry substance accumulation of root tuber. Relative growth rate changed with large ranges, higher in seeding stage, rapid decrease in stem and leaf growth stage, rose in expanding period of root tubers, and declined again in the stage of dry substance accumulation of root tuber. Crop growth rate was higher in the first and last stages, and smaller in interim stage. The growth parameters of S. ningpoensis such as relative growth rate, net assimilation rate, leaf area index, leaf area ratio, specific leaf area, crop growth rate changed along with the growth. The rule of dry matter accumulation was as follows: the dry matter increased slowly during the seeding stage and speeded up in the middle and late stages, and in dry substance accumulation of root tuber increased slower, the growth of dry matter all appeared an "S" curve, and accorded with logistic equation. Cultivation technologies of S. ningpoensis and the relevant management methods could be established according to the growth of different parts of S. ningpoensis and the characteristics of dry substance accumulation in different stage.

Genetic diversity and genetic structure of endangered wild Sinopodophyllum emodi by start codon targeted polymorphism / 中国中药杂志

<p><b>OBJECTIVE</b>Revealed the genetic diversity level and genetic structure characteristics in Sinopodophyllum emodi, a rare and endangered species in China.</p><p><b>METHOD</b>We detected the genetic polymorphism within and among six wild populations (45 individuals) by the approach of Start Codon Targeted (SCoT) Polymorphism. The associated genetic parameters were calculated by POP-GENE1.31 and the relationship was constructed based on UPGMA method.</p><p><b>RESULT</b>A total of 350 bands were scored by 27 primers and 284 bands of them were polymorphic. The average polymorphic bands of each primer were 10.52. At species level, there was a high level of genetic diversity among six populations (PPB = 79.27%, N(e) = 1.332 7, H = 0.210 9 and H(sp) = 0.328 6). At population level, the genetic diversity level was low (PPB = 10.48% (4.00% -23.71%), N(e) = 1.048 7 (1.020 7-1.103 7), H = 0.029 7 (0.012 9-0.063 1), H(pop) = 0.046 2 (0.019 9-0.098 6). The Nei's coefficient of genetic differentiation was 0.841 1, which was consistent with the Shannon's coefficient of genetic differentiation (0.849 4). Two calculated methods all showed that most of the genetic variation existed among populations. The gene flow (N(m) = 0.094 4) was less among populations, indicating that the degree of genetic differentiation was higher. Genetic similarity coefficient were changed from 0.570 8 to 0.978 7. By clustering analysis, the tested populations were divided into two classes and had a tendency that the same geographical origin or material of similar habitats clustered into one group.</p><p><b>CONCLUSION</b>The genetic diversity of samples of S. emodi is high,which laid a certain foundation for effective protection and improvement of germplasm resources.</p>