scholarly journals Genetic Diversity and a PCR-Based Method for Xanthomonas axonopodis Detection in Passion Fruit

2011 ◽  
Vol 101 (4) ◽  
pp. 416-424 ◽  
Author(s):  
C. F. Munhoz ◽  
B. Weiss ◽  
L. R. Hanai ◽  
M. I. Zucchi ◽  
M. H. P. Fungaro ◽  
...  

Xanthomonas axonopodis pv. passiflorae causes bacterial spot in passion fruit. It attacks the purple and yellow passion fruit as well as the sweet passion fruit. The diversity of 87 isolates of pv. passiflorae collected from across 22 fruit orchards in Brazil was evaluated using molecular profiles and statistical procedures, including an unweighted pair-group method with arithmetical averages-based dendrogram, analysis of molecular variance (AMOVA), and an assigning test that provides information on genetic structure at the population level. Isolates from another eight pathovars were included in the molecular analyses and all were shown to have a distinct repetitive sequence-based polymerase chain reaction profile. Amplified fragment length polymorphism technique revealed considerable diversity among isolates of pv. passiflorae, and AMOVA showed that most of the variance (49.4%) was due to differences between localities. Cluster analysis revealed that most genotypic clusters were homogeneous and that variance was associated primarily with geographic origin. The disease adversely affects fruit production and may kill infected plants. A method for rapid diagnosis of the pathogen, even before the disease symptoms become evident, has value for producers. Here, a set of primers (Xapas) was designed by exploiting a single-nucleotide polymorphism between the sequences of the intergenic 16S-23S rRNA spacer region of the pathovars. Xapas was shown to effectively detect all pv. passiflorae isolates and is recommended for disease diagnosis in passion fruit orchards.

Plant Disease ◽  
2021 ◽  
Author(s):  
Marwa Laribi ◽  
Alireza Akhavan ◽  
Sarrah M'Barek ◽  
Amor Yahyaoui ◽  
Stephen Ernest Strelkov ◽  
...  

Pyrenophora tritici-repentis (Ptr) causes tan spot, an important foliar disease of wheat. A collection of Ptr isolates from Tunisia, located in one of the main secondary centers of diversification of durum wheat, was tested for phenotypic race classification based on virulence on a host differential set, and for the presence of the necrotrophic effector (NE) genes ToxA, ToxB , and toxb by PCR analysis. While races 2, 4, 5, 6, 7, and 8 were identified according to their virulence phenotypes, PCR testing indicated the presence of ‘atypical’ isolates that induced necrosis on the wheat differential ‘Glenlea’, but lacked the expected ToxA gene, suggesting the involvement of other NEs in the Ptr/wheat interaction. Genetic diversity and the Ptr population structure were explored further by examining 59 Tunisian isolates and 35 isolates from Algeria, Azerbaijan, Canada, Iran, and Syria using 24 simple sequence repeat markers. Average genetic diversity, overall gene flow and percentage polymorphic loci were estimated as 0.58, 2.09 and 87%, respectively. Analysis of molecular variance showed that 81% of the genetic variance occurred within populations and 19% between populations. Cluster analysis by the unweighted pair group method indicated that ToxB- isolates grouped together and were distantly related to ToxB+ isolates. Based on Nei’s analysis, the global collection clustered into two distinct groups according to their region of origin. The results suggest that both geographic origin and the host-specificity imposed by different NEs can lead to differentiation among Ptr populations.


Genome ◽  
2002 ◽  
Vol 45 (6) ◽  
pp. 1175-1180 ◽  
Author(s):  
F J Massawe ◽  
M Dickinson ◽  
J A Roberts ◽  
S N Azam-Ali

Bambara groundnut (Vigna subterranea (L.) Verdc), an African indigenous legume, is popular in most parts of Africa. The present study was undertaken to establish genetic relationships among 16 cultivated bambara groundnut landraces using fluorescence-based amplified fragment length polymorphism (AFLP) markers. Seven selective primer combinations generated 504 amplification products, ranging from 50 to 400 bp. Several landrace-specific products were identified that could be effectively used to produce landrace-specific markers for identification purposes. On average, each primer combination generated 72 amplified products that were detectable by an ABI Prism 310 DNA sequencer. The polymorphisms obtained ranged from 68.0 to 98.0%, with an average of 84.0%. The primer pairs M-ACA + P-GCC and M-ACA + P-GGA produced more polymorphic fragments than any other primer pairs and were better at differentiating landraces. The dendrogram generated by the UPGMA (unweighted pair-group method with arithmetic averaging) grouped 16 landraces into 3 clusters, mainly according to their place of collection or geographic origin. DipC1995 and Malawi5 were the most genetically related landraces. AFLP analysis provided sufficient polymorphism to determine the amount of genetic diversity and to establish genetic relationships in bambara groundnut landraces. The results will help in the formulation of marker-assisted breeding in bambara groundnut.Key words: under-utilized, African legume, molecular markers.


2021 ◽  
Vol 12 ◽  
Author(s):  
Govindan Pothiraj ◽  
Zakir Hussain ◽  
Awani Kumar Singh ◽  
Amolkumar U. Solanke ◽  
Rashmi Aggarwal ◽  
...  

Though the vascular wilt of tomato caused by the species of Fusarium is globally reported to be a complex disease in certain countries, for example, India, our studies indicated that the disease is caused by either Fusarium oxysporum f. spp. lycopersici (Fol) or Fusarium solani (FS) with the Fol being widely prevalent. In assessing the genetic diversity of 14 Fol strains representing the four Indian states by the unweighted pair group method with arithmetic averaging using Inter Simple Sequence Repeat (ISSR) amplicons, the strains distinguished themselves into two major clusters showing no correlation with their geographic origin. In pot experiments under polyhouse conditions, the seed dressing and soil application of a talc-based formulation of a biocontrol treatment, TEPF-Sungal-1 (Pseudomonas putida) + S17TH (Trichoderma harzianum) + CG-A (Chaetomium globosum), which inhibited Fol, was equally effective like the cell suspensions and was even better than the fungicidal mixture (copper oxychloride-0.25% + carbendazim-0.1%) in promoting the crop growth (52.3%) and reducing vascular wilt incidence (75%) over the control treatment, despite the challenge of inoculation with a highly pathogenic TOFU-IHBT strain. This was associated with significant expressions of the defense genes, indicating the induction of host resistance by a biocontrol consortium. In field experiments on two locations, the bioconsortium was highly effective in recording maximum mean fruit yields (54.5 and 60%) and a minimum mean vascular wilt incidence (37.5%) in comparison to the untreated control. Thus, Chaetomium-based bioconsortium demonstrated consistency in its performance across the two experiments in 2 years under the two field conditions.


2007 ◽  
Vol 132 (3) ◽  
pp. 357-367 ◽  
Author(s):  
P. Escribano ◽  
M.A. Viruel ◽  
J.I. Hormaza

Cherimoya (Annona cherimola Mill.) is an underused fruit crop with a clear niche for expansion in subtropical climates. In this study, 16 simple sequence repeat (SSR) loci were used to find molecular polymorphisms among 279 cherimoya accessions from a worldwide ex situ field germplasm collection. A total of 79 amplification fragments were amplified with 16 pairs of SSR primers, with an average of 4.9 bands/SSR. Mean expected and observed heterozygosities averaged 0.53 and 0.44, respectively. The total value for the probability of identity was 4.34 × 10−8. The SSRs studied resulted in 267 different fingerprinting profiles, of which 258 were unique genotypes; the rest were putative cases of synonymies or mislabeling errors. Unweighted pair group method with arithmetic averages (UPGMA) cluster analysis indicated the relationships among the analyzed accessions, showing some specific groups related to their geographical origins. Analysis of molecular variance (AMOVA) was performed to examine the distribution of genetic variation of the 148 accessions collected from putative cherimoya origin areas in Ecuador and Peru, showing that the major variations occurred within valleys in each country. The results confirmed the usefulness of microsatellites for identification of genetic diversity and geographic origin of cherimoya and are discussed in terms of their implications for ex situ conservation of cherimoya genetic resources.


2005 ◽  
Vol 47 (5) ◽  
pp. 295-300 ◽  
Author(s):  
Jorge Fraga ◽  
Jinnay Rodriguez ◽  
Omar Fuentes ◽  
Aymé Fernandez-Calienes ◽  
Mayda Castex

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4) generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA). Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.


2020 ◽  
Vol 19 (1) ◽  
pp. 23
Author(s):  
CHAERANI CHAERANI ◽  
SRI KOERNIATI ◽  
DYAH MANOHARA

<p>ABSTRAK<br />Phytophthora capsici adalah penyebab penyakit busuk pangkal<br />batang yang paling merugikan pada lada di Indonesia dan sulit<br />dikendalikan karena dapat bertahan lama dalam tanah serta memiliki<br />keragaman agresivitas isolat luas. Pengetahuan mengenai keragaman<br />genetik strain-strain P. capsici dapat membantu perancangan strategi<br />efektif pengelolaan patogen. Penelitian ini bertujuan mengevaluasi<br />keragaman dan struktur genetik isolat-isolat P. capsici asal lada<br />menggunakan penanda RAPD. Penelitian dilaksanakan pada bulan<br />Oktober 2009 sampai April 2010 di Laboratorium Biokimia BB Biogen<br />dan Laboratorium Hama dan Penyakit Balittro. Keragaman genetik 59<br />isolat P. capsici yang berasal dari koleksi kultur tahun 1982-2009 dari 37<br />lokasi di Sumatera, Bangka, Jawa, dan Kalimantan, dikarakterisasi<br />menggunakan enam primer RAPD. Pengelompokan menggunakan<br />unweighted pair-group method with arithmatic averaging (UPGMA)<br />berdasarkan profil RAPD membagi ke-59 isolat ke dalam lima gerombol<br />utama; yang menunjukkan adanya keragaman genetik tinggi antar isolat.<br />Pengelompokan RAPD tidak berkaitan dengan asal lokasi isolat. Analysis<br />of molecular variance (AMOVA) juga menunjukkan adanya keragaman<br />genetik yang tinggi di antara isolat-isolat P. capsici, dengan ragam genetik<br />total sebesar 96% terletak di dalam masing-masing pulau (within<br />populations). Namun demikian, terdapat ragam genetik antar isolat dari<br />pulau berbeda (among populations) yang signifikan (4% ; P=0,001), yaitu<br />antar populasi di Sumatera dan Bangka dengan jarak genetik sebesar 0,081<br />(P=0,002). Ketidakterkaitan antara pengelompokan RAPD dengan asal<br />lokasi geografik isolat dan ragam genetik yang tinggi dalam satu pulau<br />dapat diakibatkan oleh terjadinya penyebaran isolat antar daerah, terutama<br />melalui bibit tanaman yang terinfestasi P. capsici. Pencegahan penyebaran<br />isolat antar pulau perlu dilakukan melalui sertifikasi bibit bebas penyakit<br />BPB dan pengembangan sistem perbenihan lokal.<br />Kata kunci: lada, penyakit busuk pangkal batang, Phytophthora capsici,<br />RAPD, keragaman genetik, struktur populasi</p><p>ABSTRACT<br />Phytophthora capsici is the causal agent of foot rot, the most<br />destructive disease of pepper in Indonesia and difficult to control .<br />Knowledge in the genetic structure of P. capsici strains can enrich<br />designing effective disease management strategies. This study was aimed<br />at analyzing the genetic variability and structure of P. capsici isolates from<br />pepper using RAPD. The study was done from October 2009 until April<br />2010 at the Biochemical Laboratory of Indonesian Center for Agriculutral<br />Biotechnology and Genetic Resources Research and Development, and the<br />Plant Pest and Disease Laboratory of the Indonesian Research Institute of<br />Spice and Medicinal Crops. Fifty-nine isolates collected from 1982 to<br />2009 from Sumatera, Bangka, Java, and Kalimantan were characterized<br />based on six RAPD markers. Unweighted pair-group method with<br />arithmatic averaging (UPGMA) clustering based on RAPD profiles<br />divided the isolates into five major cluster, which indicated high genetic<br />variability among isolates. No apparent relationship between RAPD<br />clustering and geographic origin of isolate was observed. Hierarchical<br />partitioning of genetic variation using analysis of molecular variance<br />(AMOVA) confirmed the overall high variability among isolates, with<br />96% of total genetic variance was resided among isolates within islands<br />(within populations). Nevertheless, a small (4%) but significant (P=0.001)<br />genetic variance among isolates between different islands (among<br />populations) were observed, which was detected between populations in<br />Sumatera and Bangka with genetic distance (Ф PT ) as high as 0,081<br />(P=0,002). The lack of association between RAPD clustering and<br />geographic origin as well as high genetic variance within populations may<br />have been the result of movement of isolates between locations, most<br />likely through infested plant cuttings. Use of certified and development of<br />blackpepper clones locally are required to prevent disease spread among<br />islands.<br />Keywords: black pepper, foot rot disease, Phytophthora capsici, genetic<br />diversity, RAPD, population structure</p>


2008 ◽  
Vol 133 (2) ◽  
pp. 197-203 ◽  
Author(s):  
Hua Wang ◽  
Dong Pei ◽  
Rui-sheng Gu ◽  
Bao-qing Wang

Molecular markers were used to study the genetic diversity, structure, and relationship of Juglans L. with nine populations (five from Juglans regia L. and four from Juglans sigillata Dode) in central and southwestern China. A moderate level of genetic diversity was observed at the population level with the number of effect alleles per locus (A E) ranging from 1.75 to 3.35 (average 2.39) and the proportion of polymorphic loci (P) equaling 100.0%. The expected heterozygosity (H E) within populations ranged from 0.389 to 0.687, and the average was 0.525. The proportion of genetic variation presented among populations accounted for 18.6% of the total genetic diversity. The overall gene flow (N m) among populations equaled 1.10. The unweighted pair-group method using arithmetic averages (UPGMA) clustering and the Mantel test showed that genetic distances among the nine populations are in a good agreement with their geographic distribution, supporting the viewpoint that J. regia and J. sigillata belong to one species. We suggest that the central area of the southwestern mountain regions of China could be considered as a priority for walnut genetic resource conservation.


2021 ◽  
Vol 7 (9) ◽  
pp. 713
Author(s):  
Abdelhameed Elameen ◽  
Svein Stueland ◽  
Ralf Kristensen ◽  
Rosa F. Fristad ◽  
Trude Vrålstad ◽  
...  

Saprolegnia parasitica is recognized as one of the most important oomycetes pests of salmon and trout species. The amplified fragment length polymorphism (AFLP) and method sequence data of the internal transcribed spacer (ITS) were used to study the genetic diversity and relationships of Saprolegnia spp. collected from Canada, Chile, Japan, Norway and Scotland. AFLP analysis of 37 Saprolegnia spp. isolates using six primer combinations gave a total of 163 clear polymorphic bands. Bayesian cluster analysis using genetic similarity divided the isolates into three main groups, suggesting that there are genetic relationships among the isolates. The unweighted pair group method with arithmetic mean (UPGMA) and principal coordinate analysis (PCO) confirmed the pattern of the cluster analyses. ITS analyses of 48 Saprolegnia sequences resulted in five well-defined clades. Analysis of molecular variance (AMOVA) revealed greater variation within countries (91.01%) than among countries (8.99%). We were able to distinguish the Saprolegnia isolates according to their species, ability to produce oogonia with and without long spines on the cysts and their ability to or not to cause mortality in salmonids. AFLP markers and ITS sequencing data obtained in the study, were found to be an efficient tool to characterize the genetic diversity and relationships of Saprolegnia spp. The comparison of AFLP analysis and ITS sequence data using the Mantel test showed a very high and significant correlation (r2 = 0.8317).


HortScience ◽  
2010 ◽  
Vol 45 (6) ◽  
pp. 857-862
Author(s):  
Jose A. Oliveira ◽  
Ana B. Monteagudo ◽  
Suleiman S. Bughrara ◽  
Jose L. Martínez ◽  
Ana Salas ◽  
...  

The objective of the present study was to characterize the diversity of 15 Festuca rubra accessions collected from northern Spain on the basis of amplified fragment length polymorphism (AFLP) and flow cytometry variation. Additionally, all accessions along with the cultivar Wilma (Festuca nigrescens ssp. nigrescens) were evaluated for susceptibility to one isolate of dollar spot fungus (Sclerotinia homoeocarpa F.T. Bennett) collected in Asturias. Five AFLP primer combinations of EcoRI and MseI produced 980 bands; 82.3% were polymorphic and used for analysis. The best combination of primers was EcoRI-AGC+MseI-CAG, because these displayed the highest degree of polymorphism. Jaccard's similarity coefficients between accessions varied from 0.30 to 0.63 and revealed low genetic similarity. Both the unweighted pair group method with arithmetic mean dendrogram and principal coordinate analysis distinguished two groups of accessions. Genetic variability in these accessions was not related to the geographic origin or to the agronomic data. Three accessions exhibited moderate resistance to dollar spot disease and may be valuable parent material for introducing this resistance in other susceptible cultivars.


2001 ◽  
Vol 126 (1) ◽  
pp. 7-12 ◽  
Author(s):  
F. Sanz-Cortés ◽  
M.L. Badenes ◽  
S. Paz ◽  
A. Íñiguez ◽  
G. Llácer

Forty olive (Olea europaea L.) cultivars from Valencia, Spain, were screened using random amplified-polymorphic DNA (RAPD) markers. Eighteen selected decamer primers produced 34 reproducible amplification fragments that were then used as polymorphic markers. The resulting combinations of these RAPD markers were used to discriminate 40 cultivars. Results were analyzed for similarity among cultivars and the relatedness of polymorphisms obtained between cultivars agreed with previous results using isozymes. Unweighted pair group method cluster analysis of their similarity values revealed two main groups divided according to geographic origin within Valencia. A third group, which included two Spanish cultivars from regions outside of Valencia, was clustered separately from the Valencian cultivars. RAPD technology proved useful in discriminating closely related cultivars. There was no apparent clustering of cultivars by fruit size or other morphological traits.


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