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Platycodonis Radix is the dry root of Platycodon grandiflorum of Campanulaceae, which has a variety of pharmacological effects and is a commonly used bulk Chinese medicine. In this study, the chloroplast genome sequences of six P. grandiflorum from different producing areas has been sequenced with Illumina HiSeq X Ten platform. The specific DNA barcodes were screened, and the germplasm resources and genetic diversity were analyzed according to the specific barcodes. The total length of the chloroplast genome of 6 P. grandiflorum samples was 172 260-172 275 bp, and all chloroplast genomes showed a typical circular tetrad structure and encoded 141 genes. The comparative genomics analysis and results of amplification efficiency demonstrated that trnG-UCC and ndhG_ndhF were the potential specific DNA barcodes for identification the germplasm resources of P. grandiflorum. A total of 305 P. grandiflorum samples were collected from 15 production areas in 9 provinces, for which the fragments of trnG-UCC and ndhG_ndhF were amplificated and the sequences were analyzed. The results showed that trnG-UCC and ndhG_ndhF have 5 and 11 mutation sites, respectively, and 5 and 7 haplotypes were identified, respectively. The combined analysis of the two sequences formed 13 haplotypes (named Hap1-Hap13), and Hap4 is the main genotype, followed by Hap1. The unique haplotypes possessed by the three producing areas can be used as DNA molecular tags in this area to distinguish from the germplasm resources of P. grandiflorum from other areas. The haplotype diversity, nucleotide diversity and genetic distance were 0.94, 4.79×10-3 and 0.000 0-0.020 3, respectively, suggesting that the genetic diversity was abundant and intraspecific kinship was relatively close. This study laid a foundation for the identification of P. grandiflorum, the protection and utilization of germplasm resources, and molecular breeding.
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Cultivating salt-alkali tolerant rice varieties is one of the important ways to meet the increasing food demand of growing global population. In this study, twenty-one rice germplasms with different salt-alkali tolerance were treated with six salt-alkali concentrations at germination and seedling stages. The germination potential, germination rate, shoot length, root length, root number, fresh weight of shoot and seedlings were measured. The average value of salt damage rate was used to evaluate the salt-alkali tolerance. As the salt-alkali concentration increases, the inhibition on seed germination and growth became more obvious. Upon treatment with 1% NaCl plus 0.25% NaHCO3, the salt damage rate of germination rate has the largest variation, ranging from 0% to 89.80%. The salt damage rate of each trait shows a similar trend at all concentrations. Four germplasm resources with strong salt-alkali tolerance (Dajiugu, Nippobare, Mowanggu and 02428) and 7 sensitive germplasms were screened. The salt-tolerant gene sequence of 4 salt-alkali tolerant varieties and 3 sensitive germplasms were analyzed. OSHAL3 and OsRR22 were identical among the 7 germplasms, but SKC1 and DST showed clear variations between the salt-alkali tolerant and sensitive germplasms. Besides the salt-alkali tolerant germplasm resources, this study can also serve as a reference for mining of genes involved in salt-alkali tolerance and breeding of salt-alkali tolerant rice varieties.
Subject(s)
Alkalies , Germination , Oryza/genetics , Plant Breeding , Seedlings/geneticsABSTRACT
OBJECTIVE: To study and exploit Chinese medicine Astragali Radix, the molecular markers that relates to the phenotypic traits on medicinal components of Astragali Radix and would be detected. METHODS: The genetic diversity of 43 Astragali Radix samples was analyzed with ISSR molecular marker technique and then the population genetic structure was studied through 13 selected markers. The association analysis between ISSR markers and 4 phenotypic traits of medicinal components were performed with GLM (general linear model) programs in Tassel 2.1. Certain genetic diversity was discovered among the 43 Astragali Radix samples. RESULTS: The genetic distance varied between 0.050 6 and 0.743 8, with an average of 0.274 1. Moreover, the cultivated Astragali Radix from Ningxia and Gansu province closely related to the wild Astragali Radix collected from Liupanshan town in Ningxia. On the other hand, No. 340 had the farthest relationship with other Astragali Radix. The content of polysaccharide, total saponins, total flavonoids, and Astragaloside IV ranged between 7.693-27.840 mg•g-1, 7.167-17.579 mg•g-1, 2.212-6.164 mg•g-1 and 6.070-107.920 μg•g-1, respectively. Meanwhile, linear regression analysis indicated that there was a significant positive correlation between the content of the total saponins and that of flavonoids (r=0.650 5,P=2.3×10-6<0.01), while the content of astragaloside had no significant correlation with that of polysaccharide, total saponins and total flavonoids. The population genetic structural analysis showed that the 43 samples were divided into 4 subgroups. There were total of 34 locus in 13 ISSR markers significantly associated (P<0.01) with the content of polysaccharide,total saponins, flavonoids and astragaloside , and the rate of explanation on the phenotype of related marker ranged from 8.14% to 51.39%. Among the locus, 15 were related with astragaloside content at interpretation rates above 30%, 1 with polysaccharide content an interpretation rate reached as high as 51.39% with high threshold (P<1×10-5). CONCLUSION: These results would provide supporting evidence for identification and protection of germplasm resources as well as molecular marker-assisted breeding.
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With the development of Tibetan medicine industry, the demands for Tibetan medicine were rising sharply. In addition, with the eco-environment vulnerability of Qinghai-Tibet Plateau region and the phenomenon of synonymies and homonymies in Tibetan medicine, there were a lack of resources and varieties in the clinical application of Tibetan medicine. At present, the shortage of Tibetan medicine and the inadequacy of its quality standard have become the two major problems that seriously restricted the sustainable development of Tibetan medicine industry. Therefore, it is important to develop the resources investigation and quality evaluation for Tibetan medicine, which were contribute to its resources protection and sustainable utilization. In this paper, current status of resources investigation, quality standardization, artificial breeding and germplasm resources of Tibetan medicine were presented by the integrated application of the new technologies, such as DNA barcoding and 1H-NMR, which provided a reference information for resources protection, sustainable utilization, variety identification and quality standardization of Tibetan medicine resources in Qinghai-Tibet Plateau.
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Objective: In order to correctly identify the different germplasm resources of Chuanmingshen violaceum and gain the excellent germplasm resources, SRAP molecular marker was used to analyze the genetic diversity of C. violaceum. Methods: C. violaceum was collected from seven different areas, which included 24 samples, the DNA fingerprint of C. violaceum was constructed with SRAP molecular marker, and the genetic diversity was analyzed. Results: Totally 374 bands were amplified by 37 primer pairs, of which 283 bands were polymorphic, and the polymorphic percentage was 75.67%. The SRAP-based genetic similarity coefficient of all samples ranged from 0.7267 to 0.9239, with a mean of 0.8150. The analysis of molecular variance showed higher percentages of genetic variation within population. All the accessions could be distinguished by SRAP markers. The cluster analysis results showed that 63 accessions were classified into seven groups, which were correlated with the geographical distribution of the accessions to some degree. The accessions from Langzhong and Jintang had higher diversity. Conclusion: There actually exists plentiful genetic diversity among the genetic resources of C. violaceum. SRAP marker is a useful method for analyzing the genetic diversity among C. violaceum accessions.
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Objective: To analyze the genetic diversity and genetic relationship in different cultivars of Curcumae Rhizoma (CR) and identified with molecular marker technique. Methods: Forty-two samples in four cultivars of CR germplasm resources were studied with RAPD-PCR marker. The genetic similar coefficient and genetic distance were analyzed by POPGEN32 software and clustered by UPGMA method. Results: Ten primers were selected from 90 random primers, a total of 83 loci were scored, among which 64 were polymorphic loci. The percentage of polymorphic loci was 77.5%. Genetic distance changed from 0.22 to 0.58. The dendrogram of different CR cultivars was clear. Conclusion: The abundant diversity of CR from different populations exists with significant genetic differentiation, which is the key for screening the germplasm resources of CR and the basis for breeding and biotechnological development of CR.
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OBJECTIVE: To investigate the contents of oligosaccharides in 37 batches of Morinda officinalis How samples from different habitats and germplasm resources at various ages. METHODS: HPLC-ELSD method was used to determine the contents of four oligosaccharides, i.e. sucrose, 1-kestose, nystose and 1F-fructofuranosylnystose in Morinda officinalis How at different ages from different habitats and germplasm resources. The relationships among the several factors were analyzed. RESULTS: The samples from Guangdong Province had larger amounts of sucrose, 1-kestose and 1F-fructofuranosylnystose than those from Fujian, Guangxi and Hainan Provinces. The content of nystose in the samples from Guangdong Province was similar with those from Fujian Province. The contents of sucrose and 1-kestose were the highest in the samples of 2.5 years old, while the contents of nystose and 1F-fructofuranosylnystose were the highest in the samples of 4 years old. The germplasm resources of small leaf had higher content of oligosaccharides than the large leaf germplasm in Guangdong Province, and different germplasm resources of Morinda officinalis How also had different morphological characteristics. CONCLUSION: The contents of four Morinda officinalis How oligosaccharides vary with habitat, germplasm and age. This research may provide references for the quality control of Morinda officinalis How.
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Objective: To establish an optimum reaction system suitable for ISSR analysis of Astragalus membranaceus and to analyze the genetic diversity of wild populations in Inner Mongolia. Methods: A stable and reliable ISSR reaction system was set up combining the concentration gradient of the single factor test and orthogonal test. The genetic diversity of 30 A. membranaceus populations in nine zones of Inner Mongolia was analyzed using NTSYS2.1 software. Results: The optimal ISSR reaction system (20 μL) contained 10 × PCR buffer 2.0 μL, 1.5 mmol/L MgCl2, 0.4 mmol/L deoxyribonucleotide triphosphate (dNTP), 1.5 U Taq DNA polymerase, 0.5 μmol/L primer, and 40 ng template DNA. A total of 169 amplified loci were detected by 15 ISSR primers, in which 157 loci were polymorphic loci with the percentage of 75%~100%. The genetic distance amplitude ranged between 0.242 7-0.730 8. The clustering analysis showed that 30 A. membranaceus populations could be divided into two categories, and most of them corresponded to the geographical distribution. Conclusion: ISSR-PCR reaction system for A. membranaceus is stable and reliable. Wild resources of A. membranaceus in Inner Mongolia have higher genetic diversity. The genetic relationship of the populations is correlated with its geographic location.
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OBJECTIVE: To analyze the differences in chemical composition of Pseudostellariae Radix from different germplasms. METHODS: Tweny-four batches of samples were determined by UPLC-Triple TOF-MS/MS. Characteristic peaks were extracted through treatment of mass spectrometry data including peak matching, peak alignment and noise filtering. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used for data processing. Component identification was carried out based on the mass spectrometry accurate mass and secondary mass spectrometry fragmentation information combined with database search and literature. RESULTS: The differences among samples of Pseudostellariae Radix of different germplasms were distinguishable. Twenty-one chemical compositions with significant differences were screened, among which 10 common chemical compositions showed different changing laws. CONCLUSION: This study provides experimental data for revealing the laws of Pseudostellariae Radix metabolites from different germplasms.
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Objective: To analyze the genetic polymorphism and relationship of Sophora alopecuroides from Ningxia, Gansu, Qinghai, Xinjiang, and Inner Mongolian in China, and to offer some information for the domestication and protection of wild S. alopecuroides germplasms. Methods: ISSR primers were used to analyze the genetic diversity of 22 populations of S. alopecuroides. Cluster analysis of the unweighted pair-group method with arithmetic average (UPGMA) and principal component analysis (PCA) were carried out based on the molecular data obtained, and a dendrogram was constructed. Results: Totally 433 alleles were amplified using the 51 ISSR primers. The number of allels in per primer range was 5-12, with an average of 8.49. The average percentage of polymorphic loci (PPB) was 93.30%. The Nei's gene diversity (H) and Shannon's information index (I) were 0.3351 and 0.4998, respectively. The genetic distances range was 0.1736-0.6502. Conclusion: The genetic polymorphism among 22 populations of S. alopecuroide is relatively abundant and there are no direct relationship between genetic distance and geographical distribution in them.
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Objective: Analyzing hereditary parameters and comparing the test-tube plants population of Bupleurum chinense and seeding plants in different growing periods to provide the basis for building and screening out the right way to foster superior seeds of B. chinense. Methods: The test-tube plants were transplanted into the croplands and the same type of seeds were sowed at the same time. Until the next year, after the reviving of the plants, the plants were marked and the timing measure on the data of number of tiller, number of branches, number of leaves, length of leaves, width of leaves, number of jointings, length of jointing, sum of stem thickness, plant height, and number of flowers was carried out. Then, the roots were dug up and the medical characters of the roots, for instance, length, degree of thickness, and weight of the roots were measured. Following that, the statistic analysis on the data was conducted by using the software of SPSS 20.0. Results: The relative standard deviation values of test-tube plants population of B. chinense in diverse genetic characters were all relatively small. Moreover, plants were tidy, individual difference was tiny, and genetic characters were stable. In addition, the relevance of the characters of test-tube plants in various growing periods had some differences with the seeding plants. At last, compared with the seeding plants, the test-tube plants populations of B. chinense had more obvious superiority on the yield of roots and output of seeds. Additionally, the medicinal characters and quality were quite even. Conclusion: Compared with the seeding plants, the test-tube plants population of B. chinense has the superior botanical and pharmacognosy characters. Moreover, plant tissue culture technique is an effective way to breed and enlarge the superior seeds of B. chinense.
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Objective To investigate the genetic diversity of Chrysanthemum morifolium and provide the evidence for evaluation and exploitation of C.morifolium germplasm.Methods The genetic diversities of 31 germplasm from different habitats were investigated with the technique of inter-simple sequence repeat(ISSR),which were 29 of C.morifolium,1 of C.indicum,and 1 of C.nankingense.Results Twenty-two primers were selected to produce highly reproducible ISSR bands.Among 182 amplified bands,149 showed polymorphism,the percentage of polymorphic bands(PPB) reached to 81.87%. Observed effective number of alleles(Ne),Nei's gene diversity index(He),and Shannon information index (Ⅰ) were 1.348 1,0.219 1,and 0.345 1,respectively.Conclusion ISSR Method is suitable for identification and genetic diversity analysis of C.morifolium.
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Objective For selecting and developing the excellent Bupleurum chinense to mass-produce seedlings and seeds in high quality.Methods B.chinense was picked from eight different areas,such as Lingchuan and Wanrong county in Shanxi Province,Longxi county in Gansu Province,and Shangluo in Shaanxi Province,etc.Rapid propagation was done.The testa of Zhongchai No.1 was scrapped or seeds were soaked in different phytohormones.The effects on germination rates of seeds were compared.ResultsThe optimum medium for bud propagation was B5supplemented with 1.0 mg/L 6-BA and 0.2 mg/L KT.The optimum medium for root induction of test-tube plantlets was 1/2 MS added with 0.1 mg/L NAA,0.5 mg/L IBA,and 1.0 mg/L DSC.The annual propagation coefficient of B.chinense plants was more than 1?108,and the survival rate of transplantation reached to 94%—97%.The phytohormone has little effect on seed germination of B.chinense,but scraping the testa could increase the germination percentage of seeds to 20% and shorten the germinating time greatly.Conclusion By tissue culture of excellent B.chinense,a great deal of plants and seeds could be produced in short time.By scrapping testa in a certain extent,the germination of seeds could be increased.