Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Phaeophyta) strains with RAPD and ISSR markers

Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.

Phylogenetic Analysis among Maize Exserohilum turcicum Isolates from Yunnan Province by RAPD

[ Objective ] This study aimed to investigate the genetic variation of Yunnan Exserohilum turcicum isolates from molecular level, and provide theoretical basis for E. turcicum pathogenicity virulence differentiation and effective control of disease. [Method] A total of 56 E. turc/cum isolates from some areas of Yunnan Province were analyzed by RAPD. Based on the genetic distance, a dendrogram was constructed. [Result] Ten genetic groups were formed in the dendrogram. The RAPD groups had no obvious correlation with geographic origins. Some strains from one area were closely related to some from another area. [ Conclusion] Rich ge- netic variation existed among the tested isolates.

Investigating the genetic diversity of several varieties of Iranian tomato (Solanum lycopersicum) using RAPD and ISSR molecular markers

Background:Numerous studies have demonstrated the existence of approximately 7,500 genetic tomato varieties worldwide.Hence,it is crucial to assess the genetic diversity among tomato cultivars.This study aimed to investigate the genetic diversity of selected Iranian tomato cultivars(Solanum lycopersicum)using RAPD and ISSR molecular markers.Method:Ten RAPD primers and ten ISSR primers were employed to assess the genetic diversity among 10 tomato cultivars:Matin,RFT 112,Hirad,Golsar,Raha,Hengam,Hedah,Fasa,JS12,and Emerald.Data analysis involved the UPGMA algorithm and NTYSYSpc software.Results:RAPD analysis revealed close genetic proximity between Fasa and JS12,as well as between Raha and Hadieh.Conversely,the RFT 112,Hengam,Hirad,and Emerald cultivars exhibited significant genetic diversity within this group.ISSR primer analysis identified Hengam as the most diverse variety,while Matin,Emerald,and Vibrid,as well as Raha and JS12,displayed genetic similarities with minimal observed diversity.Furthermore,the overall analysis of the cultivars using RAPD and ISSR markers indicated that Hengam exhibited the highest diversity among all the varieties.Notably,Raha and JS12 demonstrated limited diversity in this analysis.Conclusion:This research demonstrates substantial genetic diversity among the investigated tomato varieties,with Hengam displaying the highest diversity within this group.Furthermore,ISSR markers proved more effective in determining genetic diversity in tomato plants.

Genetic Diversity in Main Cultivars of Safflower in Xinjiang Uighur Autonomous Region Based on RAPD Analysis

[Objective] Study on the genetic diversity in main cultivars of safflower distributing in Xinjiang Uighur Autonomous Region by means of RAPD makers.[Method] Genomic DNAs of 29 safflower accessions from Xinjiang Uighur Autonomous Region were extracted for PCR amplification using 20 RAPD primers.[Result] Totally 156 bands were amplified,among which 144 bands were polymorphic(accounting for 92.31%),indicating that safflower is endowed with plentiful genetic diversity.Based on the DNA fingerprint,the 29 safflower accessions were grouped into four populations,the classification results may be not related with ecological regionality.[Conclusion] RAPD technique is an available tool to analyze the genetic diversity of safflower germplasm at molecular level.

RAPD analysis of genetic diversity of nine strains of Auricularia auricular cultivated in Heilongjiang Province

Polymorphism of nine strains （CF05, CF09, 29, 916, AU9, Chang10, Chang7, 8808 and AU. Japanese） of A. auricular cultivated in Heilongjiang Province were analyzed by RAPD （Random Amplication polymorphic DNA）. Thirteen primers were selected from forty PCR primers with 10bp long random primer. The results showed that nine strains of A. auricular have a high level of genetic diversity and the percentage of DNA polymorphic was 96.05. The genotypes of 9 strains of Auricularia auricular were identified by the fingerprints from primer 27 and primer 46 by RAPD analysis. The results are helpful for quickly identifying strains of A. auricular in its early breeding time, and also provides a powerful theoretic basis to differentiate strains （Auricularia auricular） whose morphology is very similar in breeding programs of edible fungus.

Analysis of Genetic Structure of Natural Populations of Castanopsis fargesii by RAPDs

Genetic diversity and population genetic structure in 188 individuals from five natural populations of Castampsis fargesii Franch. were studied by RAPD markers. Three hundred and eighty-five loci were identified with 41 oligonucleotide primers, out of which 157 loci were polymorphic and accounted for 40.78% of total genetic diversity at species level. Shannon's indices of diversity (I) and Nei's gene diversity ( h) were 0.459 7 and 0.296 at the species level, respectively. The result showed that genetic variation of C. fargesii populations mainly existed within populations. Genetic differentiation (Hsp-hpop)/Hsp estimated with Shannon's index of diversity and coefficient of gene differentiation (Gst) were 0.047 6 and 0.042 9 respectively, which were confirmed by the analysis of molecular variance (AMOVA). Therefore, it is apparent that within-population variation accounted for 94.97% and among-populations variation accounted for only 5.03% of the total genetic diversity. AMOVA also indicated that there was significant differentiation among populations as well as among individuals within a population.

Genetic Diversity in Wild Grapes Native to China Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis

The use of the RAPD technique was investigated on a set of 73 genotypes of 18 wild grape species native to China, and one interspecific hybrid, seven Vitis vinifera L. cultivars, one rootstock cultivar and one strain of V. riparia L. Genetic diversity among these grapes was investigated based on RAPD analysis. The screening of 280 decamer oligonucleotides allowed the selection of 20 primers used for the analysis. A total of 191 RAPD markers were produced from the 20 selected primers. Relationships among the 83 clones or accessions based on their genetic distances were clustered using unweighted pair_group method arithmetic average (UPGMA) analysis in a dendrogram. Twenty_two clusters which fortunately adapted to 22 grape species level were clearly resolved on the dendrogram. The 18 wild grape species native to China were grouped into ten subclusters. The largest distance was found between V. riparia L., V. vinifera L., interspecific hybrid ( V. vinifera L.× V. larbrusca L.) and the wild grapes native to China. Among the wild grapes native to China, the largest distance was found between V. hancockii Hance and the other wild species. V. qinlingensis P.C.He was the second. Large genetic variation occurred among the different flower_type clones in one species.

RAPD Analysis for Genetic Diversity of Nineteen Common and Tartary Buckwheat Varieties

[Objective]The study aimed to carry out RAPD analysis of genetic diversity of common and tartary buckwheat varieties.[Method] The genetic diversity of 19 common and tartary buckwheat varieties including the tested varieties in Guizhou region during 1999-2010 and their parents were studied using 7 primers by means of random amplified polymorphic DNA（RAPD）.[Result]A total of 149 DNA bands were obtained.In which,141 bands were polymorphic,accounting for 94.89%.Polymorphism analysis and cluster analysis showed that all varieties had their own special bands different from each other.The varieties native to Weining were close to each other,and other common buckwheat varieties were obviously different from each other.The interspecific genetic variation was the greatest;the intraspecific genetic variation of common buckwheat varieties was greater than that of tartary buckwheat varieties.[Conclusion]The RAPD fingerprints of the 19 buckwheat varieties were established in present study.

RAPD Analysis for Genetic Diversity of Varanus salvator

[Objective] The aim was to carry out RAPD analysis on genetic diversity of Varanus salvator. [Method] 20 random primers were used for PCR amplification of genomic DNA of 36 individuals of V. salvator. [Result] 10 primers could produce highly reproducible RAPD bands. A total of 2 952 DNA fragments were successfully amplified. Each individual got 82 amplified bands on average,47 of which showed polymorphism. The polymorphic locus percent was 57.32%. The genetic distance among 36 individuals ranged from 0.035 9 to 0.335 9 with an average of 0.135 9. The Nei＇s gene diversity index （H） and Shannon＇s information index （I） were 0.181 9 and 0.263 0,respectively,indicating that V. salvator had greater genetic diversity. [Conclusion] The phylogenetic tree was inferred by using UPGMA analysis,it was found that the 36 individuals could be classified as one group,and there was no obvious population differentiation.

Studies on Genetic Polymorphism of Different Biotypeswith RAPD Analysis1

In the present paper, RAPD was used to study the genetic polymorphism of fisheswith different genome combinations. Our results indicated that four of the 26 random primersproduced distinct and reproducible electrophoretic patterns which were genome-specific andcould distinguish different biotypes. This enabled us to derive a diagnostic profile, from whichwe constructed a molecular marker key for different biotypes. By the analysis of the data ofRAPD patterns, the genetic relationship was constructed with UPGMA (unweighted pair-groupmethod with arithmetical averages). Our experiments also concluded that RAPD was moresuccessful in variety identification than protein polymorphism analysis and serohematology for itstechnological simplicity and sensitivity.

Analysis of the Germplasm Resources and Genetic Relationships Among Hybrid Cymbidium Cultivars and Native Species with RAPD Markers

The random amplified polymorphic DNA （RAPD） marker was assessed to detect the genetic relationships among 48 hybrid Cymbidium cultivars from Japan, Korea, China, and USA, and 2 species of native Cymbidium. Twenty primers were screened from 100 random decamer primers, and a total of 258 DNA bands were amplified, 253 of which （98.1%） were polymorphic. The average number of polymorphic DNA bands amplified by each primer was 12.6. All cultivars were distinguishable when a number of primers were considered. Genetic similarities among the cultivars and species were estimated based on the amount of band sharing ranging from 0.364-0.817 with an average of 0.581. According to the data, a dendrogram of genetic relationship, which was constructed using the UPGMA method, showed that all the tested cultivars and native species were classified into five cluster groups with the similarity coefficient of 0.592. It revealed that the genetic relationships among tested accessions were to some extent related with their origin, flower colour, branch type, and genealogy. It further indicated that the RAPD technique is a useful tool for studying the genetic relationships among hybrid Cymbidium cultivars.

RAPD-PCR Analysis on Genetic Relationships Between Cultivars of Tree Peony

Random amplified polymorphic DNA (RAPD) was used to analyze genetic polymophism of 35 Tree Peony cultivars with 7 different color groups. Thirty four primers amplified 418 DNA fragments and 337 polymorphic bands (80.6%), including specific DNA markers for 18 cultivars that could be used to differentiate cultivars. The UPCMA method was used to analyze the genetic relationship among cultivars. The results showed that 35 Peony cultivars could be divided into 2 cluster groups when using similarity criteria of 1.5, and into 4 cluster groups when using similarity criteria of 1.0. The result confirmed that the flower color has no relation to the genetic clusters and the Tree Peony cultivars originated from the same area has close genetic relationship. Therefore, genetic background has no large effect on the genetic relationship. The sequence based on polymorphic rate from high to low was Blue groups > Yellow groups > Bark red groups > Blake groups > White groups>Green groups>Red groups.

Analysis of Genetic Diversity in Cultivated and Wild Tomato Varieties in Chinese Market by RAPD and SSR

RAPD and SSR were applied to assess genetic diversity in 61 tomato varieties from different species （Solanum lycopersicum L., hirsutum. Humb L., pimpinellifolium Miller L., chilense Dun. L., chmielenskii L., peruvianum Miller L., parvuflorum Miller L.）. 2 062 and 869 clear fragments were amplified by RAPD and SSR, respectively. On the other hand, more polymorphic products were found by SSR as compared to RAPD, i.e., 100 and 43.84%, respectively. In addition, a higher value of the average similarity coefficient and lower PIC value were reflected in RAPD （0.79, 0.407） compared to SSR （0.56, 0.687）. It can be inferred that SSR was a higher effective marker than RAPD to assess genetic diversity in tomato accessions. Similarly, the genetic base of tomato varieties in Chinese market was narrow. It is suggested that wild tomato varieties should be used to enrich the genetic base of the cultivated tomato varieties.

Population Genetic Structure and Genetic Differentiation of the Pufferfish Takifugu Rubripes and Takifugu Pseudommus Revealed by RAPD Analysis

RAPD analysis is used to assay the population genetic structure and genetic differentiation of pufferfish T. rubripes and T. pseudammus. One hundred and twenty fragments are amplified with 21 random sequence 10-mer primers. The proportion of polymorphic fragments of T. rubripes populations from the coast of China (TRC), the coast of Japan (TRJ), general population of T. rubripes both from China and Japan (TRCJ), and the population of T. pseudommus are 31.7%, 33.3%, 35.0% and 39.2 % respectively. The mean expected heterozygositiee of the four populations are 0. 116, 0. 125, 0. 126, and 0. 144, respectively. Low genetic distances (ranging from 0.0118 to 0.0309) and Fst estimates (0.020 to 0.024) among the populations in-dicated that there is no significant differentiation between T. rubripes and T. pseudommus and suggested that only one species is involved.

GENETIC DIVERSITY OF FIVE FRESHWATER MUSSELS IN GENUS ANODONTA (MOLLUSCA: BIVALVIA) REVEALED BY RAPD ANALYSIS

Unionidae are an important group of benthic freshwater species. Due to the convergence phenomenon within freshwater mussels, there is still much controversy in the classification of Chinese Unionidae. In China, most studies on freshwater mussels emphasized resource investigation, biology and morphology, while little has been done in genetics, and particularly not in population genetic structure as well as genetic diversity. In order to further understand the status of genetic diversity of different species, random amplified polymorphic DNA (RAPD) markers were used to detect genetic diversity of populations in five species of the genus Anodonta: Anodonta arcaeformis, A. arcaeformis flavotincta, A. fluminea, A. woodiana woodiana and A. w.pacifica. DNA extraction method was based on phenol-chloroform and extracted genomic DNA from the adductor muscle and mantle tissues. Sixteen random primers were used for RAPD amplification and the polymorphism of amplified loci were analyzed. The results demonstrated that the percentage of amplified polymorphic loci for various populations ranged from 34.5% to 62.8%, the mean Shannon’s genetic diversity indices ranged from 0.2021 to 0.3552, and the mean intra-population Nei’s genetic distance ranged from 0.1386 to 0.1713. In all populations of the five species, the genetic diversity for A. arcaeformis was the largest, and that of A. fluminea was the lowest. The inter-population genetic distance between A. w. woodiana and A. w. pacifica was 0.3186, so they can be considered as two sister species at the genetic angle.

Genetic Diversity Analysis of PVY Resistant Potato Varieties (Clones) by RAPD Markers

DNA fingerprints of 37 PVY resistant potato varieties （clones） were generated using PCR-based RAPD analysis. All 37 varieties were differentiated by banding patterns obtained from 17 primers that generated 164 polymorphisms. Similarity matrices were calculated with the simple matching coefficient. The genetic distance value among them was between 0.27 and 0.50, with average value being 0.35. The average link clustering algorithm was used to construct a dendrogram with Statistica. They were classified into three groups according to clustering results, and the results presented some geographic relationship. Dendrogram showed close genetic relationships between a number of varieties （clones） of similar pedigree. This study has shown that RAPD marker is a useful tool for potato variety identification, differentiation, and estimation of genetic relationships.

The comparison between allozyme and RAPD makers for the population genetic structure analysis of scallop Chlamys farreri

To compare genetic markers for population genetics analysis, allozyme electrophoresis and random amplified polymorphic DNA （RAPD） were used to detect the genetic structure of scallop Chlamysfarreri population. Thirteen enzymes （MDH, ME, IDH, GPI, PGM, PEP-LG, PEP-pp, ACP, AK, PK, AAT, SOD, EST） in three butter systems （TC, pH6.9; TMME, pH 7.4; and EBT, pHS.9） were selected and 22 loci were used for the analysis, among them 7 loci （Gpi, Pgm, Pep.m-l, Pep.pp. Aat-2, Est-2, Est-3） were polymorphic which attributed 31.82% to the total. The average of heterozygosity was 0.113 and most of the studied loci showed heterozygote deficiencies. The same specimens were investigated using 10 arbitrarily selected primers （10-base）. Twenty two of 54 RAPD fragments were polymorphic with average heterozygosity of 0.194. The result indicated that the two types of markers reflected a consistent trend in the parameter values of genetic diversity of the population, but RAPD revealed more information of genetic variation than allozyme electrophoresis.

Analysis of Genetic Relationship among Celery Germplasms by RAPD

In this study, forty celery cultivars introduced from different regions of China were used as experimental materials to verify and analyze the practicability of RAPD technology in the identification of celery cuhivars. The results showed that RAPD technology could distinguish accurately the genetic relationship among various celery cultivars as a simple and ideal DNA molecular marker technology suitable for genetic relationship analysis. To be specific, 13 RAPD primers with clear amplified bands and significantly different number of amplified bands were screened, which provided basis for the identification of genetic relationship among different celery cultivars.

Genetic Analysis of the First Female Flower Node and the First Male Flower Node in Bitter Gourd

Exploring genetic mechanism of the first female flower node and the first male flower node in bitter gourd has practical significance for formulating breeding strategy. In this article, a cross was made between CN19-1 and Thai4-6, and the F2segregation population was also constructed through F1selfing. The genetic characteristics of the first female flower node and the first male flower node were analyzed by adopting the major gene plus polygene mixed genetic model. The data analysis results showed that the first female flower node and the first male flower node were continuous distribution in the F2segregation population. E-2 model was the most suitable model for the genetic analysis of the first female flower node and the first male flower node. The additive effect values of the 2 pairs of major genes controlling the first female flower node were 2.722 and 1.862 8 respectively, the dominant effect values were-2.721 6 and-0.171 8, respectively. The additive effect value of polygene was-0.839 2, and the dominant effect value of polygene was 2.225 4. The heritability of major genes and polygene were 83.73% and 1.54%, respectively. The additive effect values of the 2 pairs of major genes controlling the first male flower node were 17.746 9 and 3.972, respectively, the dominant effect values were 5.191 6 and-3.972, respectively. The additive effect value of polygene was-20.530 5, and the dominant effect value was-4.141 4. The heritability of major genes and polygene was 92.34% and 4.7%, respectively. This study could provide a theoretical basis for bitter gourd breeding.

RAPD analysis for genetic diversity of germplasm resources of <i>Strobilanthes</i>

The 18 samples representing 18 populations of Strobilanthes cusia (Nees) O. Ktze in Fujian Province of China were analyzed with RAPD markers. Eleven primers were used, a total of 106 bands were scored and 88 of them were polymorphic. The percentage of polymorphic loci was 77.36%, Nei’s gene diversity was 0.2420, and Shannon’s index was 0.3700. The 18 populations were classified into 2 groups based on the RAPD data by the method of hierarchical cluster analysis. Most of the populations from Fujian were clustered into a group, other populations were clustered into the other group. There was a high level of genetic diversity among the populations, and the genetic differentiation was obvious among the populations from Fujian.