ABSTRACT
The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.
Subject(s)
DNA, Plant , Genetics , DNA, Ribosomal , Genetics , DNA, Ribosomal Spacer , Genetics , Magnoliopsida , Classification , Genetics , Plant Leaves , Genetics , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , RNA, Ribosomal , Genetics , RNA, Ribosomal, 18S , Genetics , Genetics , Species SpecificityABSTRACT
The study is aimed to assess the genetic diversity and genetic relationship of 18 Sarcandra glabra resources from different populations,and guide parent selection of cross breeding between these resources. The molecular marker technique ISSR was used to investigate the genetic diversity of the 18 resources. Data was analyzed by POPGEN 32, and a cluster diagram was presented by UPGMA. One hundred and ninety-eight amplified fragments were obtained using 23 ISSR primers. One hundred and eighty-four polymorphic loci were identified. Nei's genetic diversity index (h) was 0.32, Shannon diversity index (I) was 0.485 4. The genetic similarity coefficient among the resources ranged from 0.383 8 to 0.878 8 in an average of 0.661 2. The genetic distance between sample S2 and sample S18 was the farthest, so as between sample S3 and sample S18 both Nei's genetic distance was 0.957 5, The genetic distance between sample S4 and sample S5 was the closest, the Nei's genetic distance was 0.129 2,and the sample S1, S2, S3, S7, S10 were significantly different from the others based on the clustering analysis, the three groups S2 vs S3, S2 vs S6, S2 vs S18 were the best parent group selection. There was a middle level of genetic differentiation in the resources. The genetic distance between resources gives useful information to guide parent selection of cross breeding.
Subject(s)
Conservation of Natural Resources , DNA Primers , Genetics , Genetic Variation , Magnoliopsida , Classification , Genetics , Microsatellite Repeats , PhylogenyABSTRACT
<p><b>OBJECTIVE</b>To develop the characteristic chromatographic profile of Sarcandra glabra by HPLC for its quality control.</p><p><b>METHOD</b>The HPLC analysis was performed on an Agilent Zorbax Eclipse XDB-C18 column (4. 6 mm x 250 mm, 5 microm) with column temperature at 40 degree C. The mobile phase was consisted of water containing 0. 5% formic acid and acetonitrile to methanol (1:9) in gradient mode, and the detection wavelength was set at 344 nm.</p><p><b>RESULT</b>A common mode of the HPLC characteristic chromatographic profile has been establised. There were 20 common peaks , seven of which were identified, and 46 samples from different habitats were classified into five groups based on principal component cluster analysis.</p><p><b>CONCLUSION</b>The method was time-saving and can represent the chemical information and provide a scientific basis for quality control of S. glabra.</p>