scholarly journals Multilocus Sequence Analysis of Selected Housekeeping- and Pathogenicity-Related Genes in Venturia inaequalis

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 447
Author(s):  
Monika Michalecka ◽  
Joanna Puławska

The relationship between housekeeping and pathogenicity-related genes and virulence or avirulence towards the primary Malus resistance genes (R) has not been previously studied for Venturia inaequalis fungus, the causal agent of apple scab. In this study, the sequences of two housekeeping genes encoding elongation factor alpha (EF-1α) and β-tubulin and two previously unstudied effector genes of V. inaequalis from mannosidase and glucosidase families of 100 strains collected from apple cultivars with Rvi6, Rvi1, and Rvi17 and without known scab resistance genes were submitted to the analyses. Based on the phylogenetic and diversity data, as well as recombination analyses of the sequenced regions, we assessed the phylogenetic relationships and genetic structure of the pathogen within the species and the evolutionary forces that are currently acting upon this microorganism. The topology of the obtained phylograms demonstrates the lack of a relationship between the phylogenetic position of the strain and the host cultivar and the geographical origin or race of the strain. The isolates from different hosts were differentiated but did not form diagnosable, distinct phylogenetic groups. These results suggest that the analyzed genes may be too conserved to reflect the adaptation of pathogens to apple genotypes with different R genes; thus, they do not adequately reflect race discrimination. In contrast, based on variation and gene flow estimation, genetic divergence was observed among strains virulent to apple trees containing Rvi6. The results of this study confirmed a lack of free recombination between strains and demonstrated that the analyzed regions are in linkage disequilibrium and contain non-random polymorphisms associated with the strain.

2020 ◽  
Vol 56 (No. 4) ◽  
pp. 165-169
Author(s):  
Lefkothea Karapetsi ◽  
Irini Nianiou-Obeidat ◽  
Antonios Zambounis ◽  
Maslin Osathanunkul ◽  
Panagiotis Madesis

Apple scab caused by Venturia inaequalis has the most destructive effects among other phytopathogens in apple crops all over the world. The integration of resistance genes from local and domestic cultivars is a prerequisite for the efficient control of this disease and is a main target in efficient breeding approaches. Across Greece, many domestic apple cultivars are reported without deep knowledge about the presence and diversity of scab resistance genes. In this study, the presence of five resistance genes (Rvi2, Rvi4, Rvi6, Rvi8 and Rvi11) was evaluated across twenty local and domestic apple genotypes, employing twelve molecular markers closely linked to known apple scab resistance loci. Significant differences and polymorphisms among the tested cultivars were detected suggesting that some of them carry a sufficient number of resistance genes. This observed genetic diversity could be exploited in ongoing breeding approaches as a natural source of polygenic resistance against apple scab.


Author(s):  
М.I. Dulov ◽  

The use of chemical means of protection against Apple scab is associated with high material costs and causes great harm to the environment. Growing of apple varieties with stable resistance to scab (the pathogen Venturia inaequalis) will significantly limit the use of pesticides. On the territory of the Russian Federation, the Rvi6 gene is considered immune, which determines resistance to five scab races, including the most aggressive fifth. The country has scabimmune apple varieties created by scientists from VNIISPK, NCFSCHWWG, FRC named after I. V. Michurin , VSTISP and other scientific institutions. In recent years, the Rvi6 gene has been overcome by scab in many European countries, and the Rvi5 gene, which is immune to four scab races, has been overcome in Russia and Germany. When creating high-yielding apple varieties of a new generation, with good fruit flavor quality, long-term and stable resistance to scab, in addition to the rvi6 resistance gene, the most promising sources are the Rvi5, Rvi11, Rvi12, Rvi14 and Rvi15 genes. The Rvi2, Rvi4, Rvi6, Rvi7 and Rvi9 genes in the apple breeding process are best used in extended pyramids of genetic resistance to scab. This will allow you to combine several scab resistance genes that control the immune system in one apple genotype. The article describes the characteristics of DNA markers, the nucleotide sequence of primers, the size of target fragments of the PCR product, including the size of the dominant allele product for detecting Rvi genes of the Venturia inaequalis pathogen that are promising for apple breeding in varieties and hybrid material. Amplification programs were selected to identify resistance genes to various races of apple scab.


2000 ◽  
Vol 90 (1) ◽  
pp. 72-77 ◽  
Author(s):  
Jyothi Prakash Bolar ◽  
John L. Norelli ◽  
Kwai-Weng Wong ◽  
Christopher K. Hayes ◽  
Gary E. Harman ◽  
...  

The goal of this research was to improve scab resistance of apple by transformation with genes encoding chitinolytic enzymes from the bio-control organism Trichoderma harzianum. The endochitinase gene, as cDNA and genomic clones, was transferred into apple cv. Marshall McIntosh by Agrobacterium-transformation. A total of 15 lines were identified as transgenic by NPTII enzyme-linked immunosorbent assay and polymerase chain reaction and confirmed by Southern analysis. Substantial differences in endochitinase activity were detected among different lines by enzymatic assay and western analysis. Eight lines propagated as grafted and own-rooted plants were inoculated with Venturia inaequalis. Six of these transgenic lines expressing endochitinase were more resistant than nontransformed cv. Marshall McIntosh. Disease severity compared with cv. Marshall McIntosh was reduced by 0 to 99.7% (number of lesions), 0 to 90% (percentage of leaf area infected), and 1 to 56% (conidia recovered) in the transgenic lines tested. Endochitinase also had negative effects on the growth of both inoculated and uninoculated plants. There was a significant negative correlation between the level of endochitinase production and both the amount of disease and plant growth.


Plant Disease ◽  
2020 ◽  
Vol 104 (8) ◽  
pp. 2074-2081 ◽  
Author(s):  
Andrea Patocchi ◽  
Andreas Wehrli ◽  
Pierre-Henri Dubuis ◽  
Annemarie Auwerkerken ◽  
Carmen Leida ◽  
...  

Apple scab, caused by Venturia inaequalis, is a major fungal disease worldwide. Cultivation of scab-resistant cultivars would reduce the chemical footprint of apple production. However, new apple cultivars carrying durable resistances should be developed to prevent or at least slow the breakdown of resistance against races of V. inaequalis. One way to achieve durable resistance is to pyramid multiple scab resistance genes in a cultivar. The choice of the resistance genes to be combined in the pyramids should take into account the frequency of resistance breakdown and the geographical distribution of apple scab isolates able to cause such breakdowns. In order to acquire this information and to make it available to apple breeders, the VINQUEST project ( www.vinquest.ch ) was initiated in 2009. Ten years after launching this project, 24 partners from 14 countries regularly contribute data. From 2009 to 2018, nearly 9,000 data points have been collected. This information has been used to identify the most promising apple scab resistance genes for developing cultivars with durable resistance, which to date are: Rvi5, Rvi11, Rvi12, Rvi14, and Rvi15. As expected, Rvi1, together with Rvi3 and Rvi8, were often overcome, and have little value for scab resistance breeding. Rvi10 may also belong to this group. On the other hand, Rvi2, Rvi4, Rvi6, Rvi7, Rvi9, and Rvi13 are still useful for breeding, but their use is recommended only in extended pyramids of ≥3 resistance genes.


2020 ◽  
Vol 36 ◽  
pp. 5-11
Author(s):  
Mădălina Militaru ◽  
Monica Sturzeanu ◽  
Adina Iancu

Apple scab, incided by the fungus Venturia inaequalis (Cke.) Wint., is a devastating disease of apple reported from almost all apple producing Romanian areas, which causes up to 70% losses of production. Molecular markers were used for detection of scab resistance genes in 22 old and introduced apple cultivars ('Romus 3', 'Romus 5', 'Rebra', 'Rustic', 'Nicol', 'Colmar', 'Colonade', registered by Research Institute for Fruit Growing Pitesti; 'Generos', 'Iris', 'Irisem', 'Luca', 'Ciprian', 'Cezar', 'Remar', 'Valery', 'Real', registered by Research Station for Fruit Growing Voinesti, Dambovita; 'Aura', 'Starkprim', 'Ionaprim', 'Bistritean', registered by Research Station for Fruit Growing Bistrita and old cultivars: 'Domnesc', 'Cretesc'). The presence of scab resistance genes were detected using the molecular markers: AL-07 (SCAR), AM19 (SCAR), VfC for Rvi6 (Vf) gene, AD13 (SCAR) for Rvi4 (Vr1) gene, OPL19 (SCAR) for Rvi2 (Vh2) and Rvi8 (Vh8) genes and OPB12 (STS) for Rvi5 (Vm) gene. The Rvi6 gene was detected in 17 cultivars from different breeding center. The marker AD13 presents in genome of 8 cultivars, such as 'Romus 3', 'Romus 5', 'Generos', 'Iris', 'Irisem', 'Cezar', 'Remar', 'Aura'. The Rvi5 gene was revealed in 3 cultivars ('Nicol', 'Generos', 'Irisem'), only.


2011 ◽  
Vol 47 (No. 4) ◽  
pp. 156-165 ◽  
Author(s):  
J. Patzak ◽  
F. Paprštein ◽  
A. Henychová

The presence of genes for resistance to scab (Venturia inaequalis) and powdery mildew (Podosphaera leucotricha) was studied using molecular markers in a sample of 279 apple cultivars from the Czech collection of apple genetic resources. The sample comprised 37 cultivars supposed to have the Vf gene for scab resistance, 97 reference world cultivars and 145 old and local cultivars. Six PCR molecular markers for the scab resistance genes Vf, Vm, Vbj, Vr and Vh and three PCR molecular markers for the powdery mildew resistance genes Pl-w, Pl-1 and Pl-d were used. The marker for the major scab resistance gene Vf was detected in all cultivars supposed to have Vf, except in Romus 1, and in the three small-fruited cultivars Malus Evereste, Golden Gem and Hilleri. The markers of the Vr and Vh scab resistance genes were detected in 22 cultivars in combination with the marker for Vf, in 56 reference world cultivars and in 82 old and local apple cultivars. PCR molecular markers for one or two of the powdery mildew resistance genes were detected in the small-fruited cultivars Malus Evereste, Golden Gem, prof. Sprengeri and Hilleri; and in the larger fruited cultivars Hagloe Crab, Borovinka and Tita Zetei. We did not find markers for the scab resistance genes Vm and Vbj in any of the studied cultivars. They are absent also in the remaining part of the Czech collection of apple genetic resources. PCR molecular markers are useful tools for the identification of resistance genes within apple germplasm collections and can be used to increase the number of sources for disease resistance in breeding programmes.


Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 370-375 ◽  
Author(s):  
Valérie Caffier ◽  
Andrea Patocchi ◽  
Pascale Expert ◽  
Marie-Noëlle Bellanger ◽  
Charles-Eric Durel ◽  
...  

A set of differential hosts has recently been identified for 17 apple scab resistance genes in an updated system for defining gene-for-gene (GfG) relationships in the Venturia inaequalis-Malus pathosystem. However, a set of reference isolates characterized for their complementary avirulence alleles is not yet available. In this paper, we report on improving the set of differential hosts for h(7) and propose the apple genotype LPG3-29 as carrying the single major resistance gene Rvi7. We characterized a reference set of 23 V. inaequalis isolates on 14 differential apple hosts carrying major resistance genes under controlled conditions. We identified isolates that were virulent on at least one of the following defined resistance gene hosts: h(1), h(2), h(3), h(4), h(5), h(6), h(7), h(8), h(9), h(10), and h(13). Sixteen different virulence patterns were observed. In general, the isolates carried one to three virulences, but some of them were more complex, with up to six virulences. This set of well-characterized isolates will be helpful for the identification of additional apple scab resistance genes in apple germplasm and the characterization of new GfG relationships to help improve our understanding of the host-pathogen interactions in the V. inaequalis-Malus pathosystem.


2020 ◽  
Vol 1 (1) ◽  
Author(s):  
David Papp ◽  
Liqiang Gao ◽  
Ranjita Thapa ◽  
Dan Olmstead ◽  
Awais Khan

Abstract Background Breeding for resistance to apple scab (caused by Venturia inaequalis), the most devastating fungal disease of apples, relies on genetic resources maintained in germplasm collections. Methods To identify new sources of scab resistance, we evaluated 177 Malus accessions, including 27 primary and 13 hybrid Malus species from diverse geographical origins, in an orchard at Geneva, New York. We also screened a differential host set for 2 years to monitor for changes in the effectiveness of ten known scab resistance genes, which allowed us to confirm the presence of virulent pathogen races in the orchard. Results We found that ~ 37% of the wild Malus accessions and domesticated cultivars were resistant to apple scab in the field. Several of these accessions were unrelated to sources of previously known resistance genes and are promising for apple scab genetic research and resistance breeding. Cultivars carrying the Rvi6 (Vf) gene from Malus floribunda clone 821, e.g. ‘Liberty’ or ‘Florina’, remained resistant despite the breakdown of Rvi6. ‘Demir’, a Malus hybrid from Turkey, and ‘Chisel Jersey’, a traditional English hard cider cultivar, showed fewer symptoms than the Rvi6 resistant cultivar ‘Prima’. Races 1 to 7 and 9 of V. inaequalis were present in the orchard, but no scab was observed on the indicator host accessions for races 11 and 12. Conclusions Detailed and systematic screening of Malus germplasm identified resistant and moderately resistant donor accessions based on resistance reaction types. These accessions are promising for use in future genetic studies to identify novel sources of scab resistance alleles for apple breeding to develop cultivars with durable apple scab resistance.


2002 ◽  
Vol 127 (3) ◽  
pp. 365-370 ◽  
Author(s):  
Minou Hemmat ◽  
Susan K. Brown ◽  
Norman F. Weeden

The genetic basis of resistance to apple scab [Venturia inaequalis (Cke.) Wint.] in the Russian apple seedling R12740-7A (Malus Mill. sp.) was investigated. Segregation ratios obtained in crosses with susceptible cultivars suggested that at least two genes were involved, and three foliar resistance reactions (chlorotic, stellate necrotic, and pit type) were observed after inoculation. DNA markers were identified for both the stellate necrotic (Vr) and pit type (no locus designation, Vx suggested) resistance phenotypes. Comparison of resistance phenotypes with marker segregation demonstrated that only two major dominant genes were present in R12740-7A, one producing the stellate necrotic lesion and the other the pit-type lesion. The chlorotic lesion could be attributed to either unclear expression of the resistance phenotype or to susceptible genotypes not contracting the disease. These markers along with a previously published marker for Vf were used to analyze inheritance of resistance in a Vr × Vf cross in advanced breeding material. The markers identified successfully all susceptible progeny, as well as apparent escapes and individuals possessing both Vf and Vr. Thus, the markers should be useful in future screening of segregating progeny and in the pyramiding of scab resistance genes in new cultivars.


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