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1.
This work investigated the fingerprinting and phenotyping of Citrus germplasm; species selected were of historical importance belonging to Citrus limonimedica Lush. and its supposed ancestors, along with some other species of the Citrus genus. An integrated approach based on the exploitation of nuclear DNA content, morphological traits and molecular markers, such as RAPD fingerprints and ITS-based SNPs, was employed. We studied a core collection of 54 distinct accessions, including 43 genotypes of the Citrus species (18 species or supposed species) and 11 genotypes of the Poncirus genus, which was used as the reference outgroup. Morphological trait analysis and statistical analysis of DNA content and markers were useful for reconstructing a Citrus phylogeny. In particular, our experiments aimed at estimating the genetic variation within and the genetic relatedness among C limon (L.) Burm., C. limonimedica and C. medica L. to shed light on the hybrid origin hypothesis of C. limonimedica. The results of the multidisciplinary analyses allowed us to confirm a remarkable differentiation between Poncirus and Citrus genera and to highlight a close relationship among the three investigated Citrus species but a distinct difference between these three species and other species in the Citrus genus. RAPD fingerprints and ITS polymorphisms enabled us to point out a variation gradient between lemon and citron, with C. limonimedica as a possible intermediate species. Some accessions of C. medica and C. limonimedica that deviate from such a trend suggest recurrent introgression and/or hybridisation with other species of Citrus.  相似文献   

2.
The amplification efficiency and identification diversity with gene-specific primers derived from S locus glycoprotein gene (SLG) and S locus receptor kinase gene (SRK) were compared, and the geographical distribution for S haplotypes was investigated by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 72 genotypes of 5 Brassica vegetables from China. The amplification efficiency and identification diversity with class I SRK primers were generally higher than that with class I SLG in most crops tested. Class I alleles were found in total 66 genotypes and they were classified into 16, 10, 7 and 10 groups for Chinese cabbage, purple flowering stalk, cauliflower and cabbage respectively. The number of amplification accessions and identification diversity using the primers of class II SLG and SRK were quite similar. Class II alleles were detected in 55 genotypes and further grouped into one type in mustard and three in other crops. The nucleotide sequences showed high similarity between identical S haplotypes determined by reciprocal pollination and PCR-RFLP tests. It demonstrated that the PCR-RFLP analysis was feasible for identification of S alleles, and SRK should be considered as a better marker for the identification of S haplotypes than SLG. The types of S haplotypes are highly diverse in Brassica vegetables from China. Nevertheless, they were geographically limited in some Brassica vegetables, so the exchange of germplasm resources should be enhanced for breeding.  相似文献   

3.
Inter simple sequence repeats (ISSR) polymorphism in Citrus indica Tanaka (Rutaceae), an endemic and threatened wild species, was examined along with three other closely related wild taxa (C. medica L., C. latipes (Swingle) Tanaka, and C. sp. ‘Memang athur’) by analyzing 53 representative accessions sampled from North-east India. Jaccard's similarity values among 53 accessions of Citrus ranged from 0.46 to 0.97 (average = 0.75). Genetic similarity values among all the 34 accessions of C. indica were found in the range of 0.82 to 0.97 with an average of 0.90. Heterozygosity (Ht = 0.123) and Shannon's information index (I = 0.188) values estimated for C. indica revealed significantly low level of genetic variation within the species. UPGMA dendrogram grouped all 53 accessions of Citrus into four major clusters: Cluster I – C. latipes; Cluster II – C. medica; Cluster III – ‘Memang athur’ and Cluster IV – C. indica. The dendrogram placed all the 34 accessions of C. indica in five sub-clusters under Cluster IV. The placement of C. indica accessions in various sub-clusters and groups in the dendrogram was based on molecular differentiation of individual accessions rather than their geographical origin. Very low genetic diversity and destruction of its natural habitat pose serious threat to C. indica even in the Citrus Gene Sanctuary in Nokrek Biosphere Reserve (NBR) in Meghalaya. Low genetic variability, heterozygosity and Shannon's information index in C. medica, C. latipes and ‘Memang athur’ are also concerns that need to be addressed for developing appropriate strategies to conserve the genetic diversity extant in these valuable genetic resources.  相似文献   

4.
5.
A population of hybrids between Arisaema triphyllum subsp. stewardsonii and A. dracontium was investigated using molecular markers to document the hybrid origin. Total genomic DNA was extracted from A. triphyllum, A. dracontium, and the hybrids, and subjected to sequence analysis of various regions of intergenic spacer and genes of chloroplast or intergenic spacer and genes of nuclear ribosome for single nucleotide polymorphisms (SNPs). Clustering was performed using the unweighted pair group method with averages (UPGMA) tested by bootstrap (BS). Hybrid origin could not be easily confirmed in some regions. However, dendrograms constructed with a combined sequence analysis of A. triphyllum 26S ribosomal RNA gene [sequence identification (SI) 467] plus A. tortuosum phytochrome C-like (phyC) gene (SI 468) were very similar to dendrograms constructed from sequences of all regions. This suggests that selecting SI 467 and SI 468 would be practical to identify hybrid origins involving two parental species. Clustering of hybrids together with the female parent in most target regions suggests that, in Arisaema, cpDNA is considered maternally inherited.  相似文献   

6.
A cpDNA fragment of 34 genotypes belonging to Citrus and four related genera was amplified and sequenced. Chloroplast microsatellites were revealed with the length of repeats ranging from 25 to 44 bases. Other than the normal uninucleotide poly(A) repeats, a trinucleotide poly(TAA) motif was also found, the first report of such repeats in a plant chloroplast genome. According to SSR structure variations, 18 Chloroplast SSR Types (CST) were identified. The CST sequences were informative for better understanding the genetic relationships of chloroplast genomes among the analyzed genotypes and confirmed some previous hypotheses about the female parent of several hybrid accessions.  相似文献   

7.
Genetic diversity was evaluated by RAPD markers and morpho-agronomic characters for a total of 42 accessions of Barberton daisy (Gerbera jamesonii) consisting of 29 commercial and 13 wild accessions. A total of 74 polymorphic bands were obtained employing a set of 12 primer pairs. The average genetic similarity coefficient for the 42 accessions, evaluated by Jaccard index was 0.55 ranging from 0.28 to 1.00. The genetic structure found among Barberton daisy accessions was evaluated by hierarchic classification analyses and UPGMA modeling, revealing six clusters of genotypes where two of them include the wild accessions and the remaining four including commercial material, except for wild genotype number 9. Shannon (H′) index was calculated using the molecular markers to investigate the genetic variation among the Gerbera accessions and showed higher values for the commercial cluster in comparison to the values obtained for the individuals from the non-commercial cluster, namely 0.34 versus 0.27, respectively. Therefore, both calculated indices (Jaccard and Shannon) indicated the presence of higher genetic variation among commercial accessions in comparison to the cluster representing non-commercial accessions, suggesting that genetic breeding programs may focus on commercial accessions to recombine interesting genotypes with commercially important and marketing-desired characteristics.  相似文献   

8.
This study was initiated to attempt clarify the identities of taxa referred to as Prunus yedoensis that grows under natural environments in Jeju, Korea and of Yoshino cherry hybrids of cultivated origin (also recorded as P. × yedoensis) in Japan, and to understand the difference between these two taxa. P. yedoensis and other species collected from natural habitats from Jeju, Korea and cultivated materials of Yoshino cherries from Tokyo and Washington, DC, were analyzed with inter-simple sequence repeat (ISSR) markers, and sequence analysis of two chloroplast DNA (cpDNA) genes, rpl16 and trnL-trnF spacer. Depending on the source of Yoshino cherry, accessions show variations with ISSR and cpDNA. Accessions belonging to each of P. serrulata var. spontanea, P. serrulata var. pubescens, and P. sargentii were grouped closely to P. yedoensis and Yoshino cherry accessions. However, two Yoshino cherry accessions that include ‘Akebono’ showed the same rpl16 haplotype of A and A at the position of 113 and 206, respectively, which were found in 4 out of 16 P. yedoensis accessions. Twelve accessions of P. yedoensis and 11 other Yoshino cherries showed rpl16 haplotype of T and A at these positions. P. yedoensis native to Korea can be considered different from Yoshino cherry of hybrid origin from Japan based on ISSR markers and rpl16 haplotypes. Therefore, it may be concluded that the Korean taxon currently referred to as P. yedoensis can be considered indigenous and sufficiently distinct to warrant recognition as a distinct entity.  相似文献   

9.
The genetic relationships among 63 melon (Cucumis melo L.) genotypes collected from various regions of Turkey were determined by comparing their molecular ISSR, SRAP, and RAPD markers with those of 19 foreign melon genotypes to investigate the taxonomic relationships and genetic variation of Turkish melon germplasm. Total 162 polymorphic markers (69, 18, and 75 obtained from ISSR, SRAP, and RAPD primers, respectively) were used to define the genetic similarity among the melon genotypes by dendrogram or two and three dimensional scalings. The average similarity (SM coefficient) between any two pairs of accessions examined as estimated by molecular variation was 0.73 ± 0.48. Within-group genetic similarities ranged between 0.46 and 0.96. Related genotypes or genotypes collected from similar regions were partitioned to similar clusters. Southeastern Anatolian genotypes were distinctly apart from group inodorus and group cantalupensis (sweet) genotypes. This reinforced the position of Turkey in the secondary genetic diversity center of melon. The genetic diversity among Turkish genotypes (H = 0.28 and I = 0.42) was only a little less than that of the world accessions (H = 0.30 and I = 0.45). On the other hand, the percentage of polymorphic loci among Turkish melon genotypes (90.7%) was even higher than that of the world accessions (87.6%).  相似文献   

10.
The responses to root zone salinity (0, 25 and 50 mM NaCl) by 40 citrus rootstock genotypes introduced from the People's Republic of China, measured as shoot chloride (Cl) and sodium (Na+) ion accumulation, growth and dry matter accumulation, were investigated under glasshouse conditions. Two experiments, one using aerated nutrient solutions and the other irrigated sand cultures, were conducted with plants grown from rooted cuttings taken from representative trees of different mandarin (Citrus reticulata Blanco and C. erythrosa Hort. ex Tan.), yuzu (C. junos Sieb. ex Tan.), Ichang papeda (C. ichangensis Swing.), sour orange (C. aurantium L.) and trifoliate orange (Poncirus trifoliata (L.) Raf.) genotypes. Two standard genotypes, viz. Rangpur lime (Citrus x limonia Osbeck.) and William's trifoliate orange were included in the experiments as reference controls.  相似文献   

11.
The trnL (UAA) intron and the intergenic spacer between the 3′ exon of trnL (UAA) and trnF (GAA) sequences were used as genetic markers for differentiating Ficus carica cultivars and establishing refined genetic relationships. The study was based on 20 fig cultivars, collected from south and centre of Tunisia. Since, the intron was thought to be more variable among close relatives than is the chloroplast spacer. The size of these non-coding regions varied from 554 to 589 and from 989 to 1022 bases pairs for the intron and the combined sequences correspondingly. The average of GC content was 33.9% and 34.6% in the intron and the combined intron and spacer respectively. High values of A + T contents were detected in both data sets and may explain the high proportions of transversions founded. The observed variation pattern of plastid DNA provides evidence of an important genetic diversity. The overall transition/transversion bias (R) was 0.202 in the intron and 0.27 in the combined regions. The RI index of 0.592 indicates that these combined sequences have clearly more homoplasy then the intron (RI = 0.705) and spacer (RI = 0.777) sequences separately. Phylogenetic trees were generated based on maximum parsimony (MP) and neighbor-joining (NJ) analysis of the chloroplast sequences data. Results proved that a typically continuous genetic diversity characterizes the local fig germplasm. In fact, relationships inferred from the cpDNA analysis suggest several clades, which do not show geographical or tree sex correspondence. Although the level of apparent diversity is considerable, we may conclude that non-coding regions of chloroplast genome provide a new and practical opportunity to evaluate genetic diversity and to discriminate fig cultivars. Revealed cytoplasmic DNA markers are reliable to elaborate a molecular data base to conduct management and breeding programs on local fig germplasm.  相似文献   

12.
AFLP technique was applied to assess genetic diversity among 44 common bean accessions that included 6 exotic accessions, 15 Indian land races and 23 released varieties. Eight AFLP primer pairs were used that produced 820 products of which 698 were polymorphic (85.12%). Wide variations were observed among all the accessions for the number of amplification products, percent polymorphism and average polymorphism information content (PIC). The Jaccard's similarity indices (J) based on the AFLP profiles were subjected to UPGMA cluster analysis. The dendrogram generated revealed seven major groups. Seventeen out of 23 released varieties were restricted to clusters VI and VII. The value of r = 0.934 in Mantel's test for cophenetic corrlelation applied to the cluster analysis indicated the high fitness of the accessions to a group. The germplasm used in the present study had narrow genetic base, although moderate to high genetic diversity was observed. The details of diversity analysis and the potential use of Indian common bean accessions in common bean breeding programme are provided in the present study.  相似文献   

13.
Banana (Musa spp.) is one of the most important crops in the world. In this study, 216 banana accessions, 184 from the National Banana Germplasm Collection of China (NBGCC) and 32 from the International Network for the Improvement of Banana and Plantain (INIBAP), were used to determine the genome composition of banana plants in these collections and to estimate their genetic diversity. The genome composition was examined using PCR-RFLP markers. The molecular data for all but one accession (ITC 1231) from INIBAP were in agreement with the initial records based on phenotypic characteristics. Microsatellite (SSR) markers were used to investigate the genetic variability and relationships among these banana accessions. Ten of the 47 primer pairs tested consistently produced reproducible and discrete fragments. We identified a total of 92 alleles, ranging from 5 to 15 per locus. The genetic similarity between the accessions ranged between 0.1 and 1, when estimated using Jaccard's coefficient. The UPGMA method based on genetic similarities, grouped the NBGCC accessions according to those containing the ‘A’ and ‘B’ genomes. However, this analysis could not separate all the accessions, especially the somatic mutations, using the primers in this study. These data indicated that limited genetic variation exists within these accessions and the collections of NBGCC should include a much wider range of banana plant material.  相似文献   

14.
For successful conservation and domestication of a species, evaluation of its genetic diversity by different markers is important. Morphological characteristics, phytochemical variation and random amplified polymorphic DNA (RAPD) profiles were generated in different accessions of Podophyllum hexandrum in order to determine the genetic diversity. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity among the accessions used in the study. There was also high diversity in the concentration of marker compounds in the collected samples as revealed by HPLC analysis. It is shown that the approaches used in the work successfully discriminate between the accessions of this species and thus they constitute interesting tools to analyze molecular, biochemical and phenotypic diversity within this species. Similarity measurement using UPGMA followed by cluster analysis resulted in formation of many groups based on geographical distribution that generally reflected expected trends between the genotypes. There were also some important exceptions like PW-S, an accession from Wastoorwan, Khrew showing close resemblance to PG-S and PG-B collected from Gulmarg but grown at two different gene banks at Srinagar and Bonera. Further an accession PSH-B from Keller was significantly diverse from the rest of the native genotypes phytochemically, morphologically and at molecular level. RAPD data analysis was found to be significant predictor of phytochemical markers in cultivated P. hexandrum germplasm. Twelve accessions grown in gene bank repository were subjected to RAPD analysis and were assessed for content of podophyllotoxin and podophyllotoxin β-d-glycoside by HPLC. Individual regressions of podophyllotoxin and podophyllotoxin β-d-glycoside by RAPD analysis against HPLC has been found to determine linear values. Strong correlation and a strong association of values of the phytochemical variables and the DNA polymorphism data has been recorded.  相似文献   

15.
We isolated and characterized 5 MADS-box cDNA clones of CitMADS1, CitMADS3, CitMADS5, CitMADS6 and CitMADS8 from fruit tissues of Satsuma mandarin (Citrus unshiu Marc.). Sequence analysis revealed that they have high sequence identities with known MADS-box genes and that their predicted proteins possess the general structural features of the M domain and the K domain. In a phylogenetic tree with 24 known Arabidopsis MADS-box genes, 5 Citrus MADS-box genes were classified into 4 clades of the MADS-box gene with independent gene functions. Their broad expression profiles in the Citrus life cycle suggested that they play roles in flower and fruit development. Most of them, except CitMADS1 and CitMADS3, are expressed in seedlings before the phase transition from vegetative to reproductive growth. CitMADS1 and CitMADS3 are not expressed in vegetative organs and could serve as a molecular marker for reproductive development.  相似文献   

16.
A rapid and reliable method to accurately identify hybrids at an early age is essential to the success of Leucadendron breeding programs because identification based on morphology can be difficult or impossible when the seedlings are young. DNA based PCR-RFLP and random amplified microsatellite polymorphism (RAMP) markers were developed for this purpose. Unexpected non-parental fragments appeared during the PCR-RFLP analysis of the nuclear ITS region of L. uliginosum 05 × L. procerum 04 hybrids. Mixing DNA from both parents in a single PCR also produced the non-parental fragment, suggesting that PCR recombination had introduced a novel restriction site into the products from the hybrids. Sequencing of individual amplified ITS products from the hybrids confirmed this conclusion. To avoid this complication, RAMP markers were developed for accurate hybrid identification in Leucadendron. RAMP analysis generated a considerable number of polymorphic products, and showed more discrimination in identifying Leucadendron hybrids than did PCR-RFLP.  相似文献   

17.
Knowledge of genetic divergence among inbred lines is essential for cross breeding. The objectives of this study were to (1) analyze the level and character of genetic diversity in C. moschata accessions, and (2) classify the genetic divergence among inbred lines in C. moschata to assist in selection of parent genotypes for genetic improvement. Twenty agronomic characters were investigated and rich diversities were demonstrated among 39 inbred lines of C. moschata from China. Various degree correlations among these characters made it possible to summarize the diversities of the twenty characters into 3 major principal components: leaf, fruit and flesh quality factor. Forty-one inbred lines of pumpkin were clustered into four groups based on principal component data, which is more distinct for classification that based on the original data of the 20 characters. However, parent inbred lines whose hybrids displayed significant heterosis in fruit weight, soluble solid and fruit shape were located in different clusters or sub-clusters based on standardized original data. It was suggested that genotypes in the same clusters may represent members of one heterotic group.  相似文献   

18.
Eighty-six triploid Citrus plants were recovered from interploid crosses between a natural tetraploid selection of the tangerine ‘Dancy’ (Citrus reticulata Blanco, cultigroup ‘Tangerine’), used as the pollen parent, and two seedy selections of diploid mandarins (C. reticulata Blanco) as well as one clementine (Citrus clementina Hort. ex Tan.): ‘Fortune’ mandarin, ‘Wilking’ mandarin and ‘Monreal’ clementine. Flow cytometric analysis was used for screening the triploid plantlets and the ISSR-PCR technique was used to characterize the obtained triploids through a double approach, confirming the hybrid nature of the offspring and allowing the analysis of the genetic pool obtained. Selection among triploid genotypes generated from elite seedy parents will give us a better chance to obtain superior mandarin cultivars characterized by true seedlessness.  相似文献   

19.
Traditional apple cultivars from Bosnia and Herzegovina (B&H), potentially diverse due to specific geographic location and history of the country, represent a possible source of valuable traits for future breeding efforts and sustainable fruit growing. A total of 39 accessions, 24 traditional B&H cultivars and 15 modern international cultivars, maintained at the ex situ apple collection “Srebrenik” in Northeast Bosnia were, investigated using 10 SSR (simple sequence repeats) markers and 23 morphologic characteristics. All the used primer pairs manage to amplify clearly distinguishable and highly polymorphic SSR alleles, in average 10.4 alleles per locus. More than two different alleles per locus were detected for seven accessions (five traditional B&H cultivars and two international cultivars). Forty one unique alleles were exclusively present within the B&H cultivars, while seven unique alleles were only detected within international cultivars. The differentiation between traditional B&H and international cultivars (Fst = 0.060; P < 0.0001) was significant, also confirmed by analyses of molecular variance (AMOVA) (fCT = 0.092; P < 0.001). Cluster analyses of 39 apple accessions, based on 10 SSR loci, revealed that only two traditional B&H cultivars grouped tightly with international cultivars (Ljepocvjetka and Bobovec Jon), while the rest formed separate clusters. Multivariate analyses of variance (MANOVA), nonparametric multivariate analyses of variance (NPMANOVA) and analyses of similarity (ANOSIM) showed statistically significant difference in morphologic characteristics between traditional B&H cultivars and the international cultivars. Cluster analyses of 39 apple accessions, based on the morphologic data, displayed less differentiation between traditional and international accessions, in comparison to the cluster analyses based on molecular data. No correlation between the molecular and morphologic data set was detected using the Mantel test. Many of the morphologic characteristics which have been analyzed in this study have significant commercial importance, we can assume that unlike the microsatellites these traits have been under agronomic selection pressure.  相似文献   

20.
Genetic diversity and relatedness of 23 yardlong bean (Vigna unguiculata spp. sesquipedalis) accessions and 7 accessions of a hybrid between cowpea (V. unguiculata spp. unguiculata) and yardlong bean (dwarf yardlong bean) in Thailand were estimated using morphological characters, simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers. In addition, two mungbean (Vignaradiata (L.) Wilczek) and two blackgram (Vigna mungo (L.) Hepper) accessions were also used as outgroup species for molecular analysis. Five morphological characters were diverse among most accessions. However, five groups of 2–3 accessions could not be distinguished from one another based on these morphological characters alone. Unweighted pair-group arithmetic average (UPGMA) analysis of these characters separated these 30 accessions into 2 major groups; the yardlong bean group and the dwarf yardlong bean group. Eleven of the sixteen SSR primers yielded clear SSRs, ten of which were polymorphic (90.91% polymorphism), detecting a total of 54 alleles with an average of 4.91 alleles per locus. These 10 polymorphic SSR markers successfully distinguished 28 yardlong bean and dwarf yardlong bean accessions. The polymorphic information content (PIC) among genotypes varied from 0.251 to 0.752 with an average of 0.597. Among the 16 ISSR primers used, a total of 312 ISSR fragments were amplified for these three Vigna species, revealing the polymorphism percentage of 91.03%. The average ISSR PIC value (0.197) with the range of 0.137–0.276 was lower than that of SSR. Nevertheless, the average marker index of this multilocus marker was 3.495, which was higher than that of SSR (0.669), owing to the differences in the effective multiplex ratio. In addition, Mantel test cophenetic correlation coefficient was higher for ISSR (0.566) than that of SSR (0.198). These results indicated higher efficiency of ISSR for estimating the levels of genetic diversity and relationships among yardlong beans and dwarf yardlong beans in this study. Pair-wise coefficients of SSR- and ISSR-based genetic similarity among all yardlong bean and dwarf yardlong bean accessions averaged 0.87 and 0.91, respectively, suggesting a narrow genetic base that emphasizes the need to broaden genetic diversity to ensure continued breeding success. Clustering of genotypes within groups was not similar when SSR and ISSR derived dendrograms from UPGMA analysis were compared. It appeared that ISSR was the most effective marker system in determining the genetic variability and relationships among yardlong bean and dwarf yardlong bean accessions and differentiating three Vigna species. In addition, ISSR was also most useful for variety identification since all 30 yardlong beans and dwarf yardlong bean accessions can be effectively distinguished by only four ISSR primers with the highest PIC values.  相似文献   

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