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1.
J.P.T. Valkonen K. Wiegmann J.H. Hämäläinen W. Marczewski & K.N. Watanabe 《The Annals of applied biology》2008,152(1):121-130
The tuber‐bearing wild potato species, Solanum stoloniferum, carries a dominant gene, Rysto, which confers extreme resistance (ER) to Potato virus Y (PVY). This gene was introgressed to cultivated potato germplasm (Solanum tuberosum) using accessions of S. stoloniferum maintained in European gene banks. It is mainly used in potato breeding programmes in Europe. Rysto was recently mapped to potato chromosome XII. However, in this study, a different accession of S. stoloniferum (PI275244; Haw1293) was used as a female parent in a cross to obtain a diploid (2n = 2x = 24) potato population of 112 F1 genotypes. From this accession, ER to PVY has been introgressed to the potato breeding programmes at the International Potato Center (Peru). As expected, ER to PVY was inherited in a dominant, monogenic fashion in the F1 population. Marker‐specific choices of DNA polymerase and adjustments of PCR conditions were made to optimise marker detection. The corresponding gene (Rysto) was mapped to the chromosome XII using the previously described and new cleaved amplified polymorphic sequence (CAPS) markers, which are based on the restriction fragment length polymorphism loci GP122 (six markers) and GP269 (one marker), and the simple sequence repeat marker STM0003. Four GP122‐based CAPS markers and STM0003 detected the same genotypes expressing ER to PVY. Because of a few recombinants, that is ER genotypes lacking the markers and the genotypes that react with necrosis but contain the markers, the marker distance from Rysto was estimated as 15.2 cM in this F1 population. However, the distance may be less if necrosis was considered an altered response also controlled by Rysto. The markers also specifically detected independent European potato cultivars that express ER to PVY derived from S. stoloniferum. Phylogenetic analysis of the sequences amplified from the GP122 locus of S. stoloniferum and potato cultivars further confirmed that the Rysto gene from independent accessions of S. stoloniferum can be selected using the same markers and the protocols described in this study. 相似文献
2.
H. Barker 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(8):1258-1262
Extreme resistance to the potato V potyvirus (PVV) was found in four potato cultivars that contain Ry genes from Solanum stoloniferum. When plants of these cultivars, were inoculated by grafting in shoot tips from PVV-infected tomato plants, necrotic symptoms
developed in some cultivars, although a full hypersensitive reaction was not elicited, while other cultivars were symptomless.
PVV replication was not detected in any of the inoculated plants by ELISA, an infectivity assay of leaf extracts by manual
inoculation to Nicotiana benthamiana indicator plants, or by ‘return grafting’ of shoot tips taken from newly developed shoots of the potato plants to virus-free
indicator plants of tomato. These methods readily detected PVV infection in inoculated plants of cv ‘Flourball’, which does
not contain an Ry gene and is susceptible, and in cvs ‘Maris Piper’ and ‘Dr Macintosh’, which contain gene Nv conditioning a hypersensitive reaction to inoculation. One of the Ry-containing cultivars, ‘Barbara’, has been previously shown to contain two genes that control extreme resistance, defined
as no viral replication in intact plants, to the potyviruses potato viruses Y and A (PVY and PVA). These genes are: Ry
sto
, which conditions resistance to PVY and PVA, and gene Ra, which conditions resistance to PVA only. It was found that in genotypes from a progeny of the cross ‘Barbara’ (Ry
sto
/Ra)בFlourball’ (ry/ra), extreme resistance to PVV segregated with gene Ry
sto
. It is proposed that either gene Ry
sto
conditions broad-spectrum extreme resistance to the distinct potyviruses PVY, PVA, and PVV or that Ry
sto
represents a family of genetically closely linked genes each controlling resistance to a specific virus.
Received: 27 December 1996 / Accepted: 9 June 1997 相似文献
3.
Song YS Hepting L Schweizer G Hartl L Wenzel G Schwarzfischer A 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,111(5):879-887
The inheritance of extreme resistance to PVY (Ry
sto) by a single dominant locus was confirmed by obtaining a 1:1 segregation ratio in a virus inoculation test with 28 resistant
(Ryry) to 29 susceptible (ryry) anther culture-derived dihaploid lines (2n=2x=24) from cv. “Assia” (2n=4x=48) having extreme resistance derived from Solanum stoloniferum in simplex constitution (Ryryryry). Twelve Ry
sto markers selected in AFLP assays using bulked segregant analysis were applied to 106 tested potato cultivars from Germany,
The Netherlands and Poland and 19 potato cultivars were identified by these markers as extremely resistant to PVY in alignment
with phenotypic data. The locus for extreme resistance (Ry
sto) to PVY was mapped on chromosome XII co-segregating with the SSR marker STM0003. The utility of anther-culture derived dihaploid
potatoes for genetic marker development was demonstrated. Marker transferability from diploids to tetraploids provides an
optimistic potential for marker-assisted selection in potato breeding programs. 相似文献
4.
G. Brigneti J. Garcia-Mas D. C. Baulcombe 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(2):198-203
Ry
sto
is a dominant gene which confers resistance to potato virus Y (PVY) in potato. We have used bulked segregant analysis of
an F1 tetraploid potato population to identify three AFLP markers linked to and on either side of Ry
sto
. The tomato homologue of one of these AFLP markers was assigned to linkage group XI by analysis of an F2 mapping population of tomato, suggesting that Ry
sto
is also on chromosome XI of the potato genome. This map position was confirmed by the demonstration that Ry
sto
was linked to markers which had been previously mapped to chromosome XI of the potato genome. Four additional AFLP markers
were identified that were closely linked to Ry
sto
in a population of 360 segregating progeny of a potato cross between a resistant (Ry
sto
) and a susceptible parent. Two of these markers were on either side of Ry
sto
, separated by only a single recombination event. The other two markers co-segregated with Ry
sto
.
Received: 29 July 1996 / Accepted: 30 August 1996 相似文献
5.
Bogdan Flis Jacek Hennig Danuta Strzelczyk-yta Christiane Gebhardt Waldemar Marczewski 《Molecular breeding : new strategies in plant improvement》2005,15(1):95-101
A novel locus for extreme resistance to Potato virus Y (PVY), Ry-fsto, was identified on potato chromosome XII. The gene Ry-fsto has been introgressed from the wild potato species Solanum stoloniferum. Inheritance of Ry-fsto in the tetraploid potato population Rysto was consistent with the model of a single, dominant gene. Bulked segregant analysis identified an ISSR (inter-simple sequence repeat) marker UBC 857980 linked to Ry-fsto. This marker mapped to linkage group XII of a reference potato RFLP (restriction fragment length polymorphism) map. Chromosome XII specific RFLP markers were converted into PCR-based STS and CAPS markers and tested for linkage with Ry-fsto in the population Rysto. CAPS marker GP122718 was tightly linked to the resistance gene and was successfully used to identify Polish and German cultivars expressing extreme resistance to PVY. This indicates that the source of Ry-fsto has been widely utilized in various potato breeding programs and can be monitored by a diagnostic marker in marker-assisted selection. 相似文献
6.
Mapping and marker-assisted selection for a gene for extreme resistance to potato virus Y 总被引:7,自引:1,他引:6
J. H. Hämäläinen K. N. Watanabe J. P. T. Valkonen A. Arihara R. L. Plaisted E. Pehu L. Miller S. A. Slack 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(2):192-197
The chromosomal location of the major gene Ry
adg
controlling extreme resistance to potato virus Y (PVY) in Solanum tuberosum subsp. andigena was identified by RFLP analysis of a diploid potato population. A total of 64 tomato and potato RFLP markers were screened
with the bulked segregant analysis (BSA) on segregants extremely resistant, hypersensitive or susceptible to PVY. Four markers
TG508, GP125, CD17 and CT168 at the proximal end of chromosome XI showed close linkage with extremely resistant phenotypes.
TG508 was identified as the closest marker linked with the Ry
adg
locus with the maximum map distance estimated as 2.0 cM. The 4 markers linked with the Ry
adg
locus were tested on independent tetraploid and diploid potato clones and were subsequently found useful for marker-assisted
selection for plants containing Ry
adg
.
Received: 5 July 1996 / Accepted: 19 July 1996 相似文献
7.
Predicted kinase-3a motif of a resistance gene analogue as a unique marker for virus resistance 总被引:2,自引:1,他引:1
V. A. Sorri K. N. Watanabe J. P. T. Valkonen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(1-2):164-170
A DNA fragment (ADG2, 310 bp) 77% homologous to the gene N (resistance to tobacco mosaic virus in Nicotiana glutinosa) and 53% homologous to RPP5 (resistance to Peronospora parasitica in Arabidopsis thaliana) was amplified by PCR from the diploid potato genotype 2x(v-2)7 that carries the gene Ry
adg
located on chromosome XI and conferring extreme resistance to potato virus Y(PVY). Sequence comparison revealed that ADG2
spans a region corresponding to the predicted kinase-2 and kinase-3a motifs in N and RPP5. One of the 12 nucleotide differences detected between ADG2 and a homologous fragment from a PVY-susceptible potato genotype
was located within the predicted kinase-3a motif. This single nucleotide substitution of G→C, resulting in an amino-acid substitution
Ser→Thr, abolished the BbvI recognition site of ADG2, which was shown to distinguish all tested potato genotypes carrying Ry
adg
from those lacking this gene, irrespective of the genetic background and ploidy level. This PCR-based resistance marker,
developed using a resistance gene analogue as a target, is the first example of a PCR-based marker that is generally applicable
for selection of an economically important trait in potato.
Received: 28 November 1998 / Accepted: 28 December 1998 相似文献
8.
Gebhardt C Bellin D Henselewski H Lehmann W Schwarzfischer J Valkonen JP 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2006,112(8):1458-1464
Closely linked PCR-based markers facilitate the tracing and combining of resistance factors that have been introgressed previously into cultivated potato from different sources. Crosses were performed to combine the Ry
adg
gene for extreme resistance to Potato virus Y (PVY) with the Gro1 gene for resistance to the root cyst nematode Globodera rostochiensis and the Rx1 gene for extreme resistance to Potato virus X (PVX), or with resistance to potato wart (Synchytrium endobioticum). Marker-assisted selection (MAS) using four PCR-based diagnostic assays was applied to 110 F1 hybrids resulting from four 2× by 4× cross-combinations. Thirty tetraploid plants having the appropriate marker combinations were selected and tested for presence of the corresponding resistance traits. All plants tested showed the expected resistant phenotype. Unexpectedly, the plants segregated for additional resistance to pathotypes 1, 2 and 6 of S. endobioticum, which was subsequently shown to be inherited from the PVY resistant parents of the crosses. The selected plants can be used as sources of multiple resistance traits in pedigree breeding and are available from a potato germplasm bank. 相似文献
9.
H. Barker 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(5-6):710-716
Extreme resistance in cultivated potato (Solanum tuberosum) to potato viruses Y and A (PVY and PVA) conditioned by the presence of Ry genes introduced from Solanum stoloniferum was described by Cockerham (1970). Cockerham detailed a number of genes which controlled a variety of reactions, including extreme resistance to both viruses (i.e. little or no visible reaction of plants and no viral replication following graft and manual inoculation) controlled by gene Ry
sto. In the present study, cvs Pirola and Barbara, which contain a Ry gene, were found to have extreme resistance to PVY isolates from the ordinary (PVY°), veinal necrosis (PVYN) and potato tuber necrotic ringspot (PVYNTN) subgroups, and PVA. The inheritance of this phenotype was examined in seedling progenies obtained by crossing Barbara and Pirola with susceptible cultivars. Segregation data for resistance to PVY and PVA in a progeny involving cv Pirola best fitted a genetical model of one gene controlling extreme resistance to both PVY and PVA, although the possibility that there are two genes, each controlling resistance to one virus but closely linked, cannot be excluded. Segregation data from progenies involving cv Barbara best fitted a genetical model in which there are two independent genes, one controlling extreme resistance to PVA and PVY and a second gene controlling extreme resistance to PVA but not to PVY. This previously unrecognised gene conferring extreme resistance to PVA only, should be given the notation Ra in keeping with nomenclature used for other resistance genes. 相似文献
10.
J. H. Hämäläinen V. A. Sorri K. N. Watanabe C. Gebhardt J. P. T. Valkonen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(8):1036-1043
The gene Ry
adg
that confers resistance to potato Y potyvirus (PVY) in the cultivated potato [Solanum tuberosum subsp. andigena, line 2x(v-2)7] is located on chromosome XI in a segment that contains three other known resistance genes in other syntenic solanaceous
species. One of them is the gene N that controls resistance to tobacco mosaic tobamovirus in tobacco and has previously been isolated and sequenced. Three sequence-related,
resistance gene-like (RGL) DNA fragments (354–369 bp) highly homologous to the gene N were PCR-amplified from the potato line 2x(v-2)7. Two RGL fragments (79 and 81% homologous to the N gene) co-segregated with Ry
adg
among the 77 F1 progeny tested. These RGLs may originate from a resistance gene family on chromosome XI. The potato line
2x(v-2)7 also expressed resistance to potato A potyvirus (PVA), which was controlled by another locus on chromosome XI mapped
ca. 6.8 cM distal to Ry
adg
.
Received: 18 December 1997 / Accepted: 30 December 1997 相似文献
11.
Iga Tomczyńska Florian Jupe Ingo Hein Waldemar Marczewski Jadwiga Śliwka 《Molecular breeding : new strategies in plant improvement》2014,34(2):471-480
Potato virus Y (PVY, Potyvirus) is the fifth most important plant virus worldwide in terms of economic and scientific impact. It infects members of the family Solanaceae and causes losses in potato, tomato, tobacco, pepper and petunia production. In potato and its wild relatives, two types of resistance genes against PVY have been identified. While Ry genes confer symptomless extreme resistance, Ny genes cause a hypersensitive response visible as local necrosis that may also be able to prevent the virus from spreading under certain environmental conditions. The potato cultivar Sárpo Mira originates from Hungary and is highly resistant to PVY, although the source of this resistance remains unknown. We show that cv. Sárpo Mira reacts with a hypersensitive response leading to necrosis after PVYNTN infection in detached leaf, whole plant and grafting assays. The hypersensitivity to PVYNTN segregated amongst 140 individuals of tetraploid progeny of cvs. Sárpo Mira × Maris Piper in a 1:1 ratio, indicating that it was conferred by a single, dominant gene in simplex. Moreover, we identified five DNA markers linked to this trait and located the underlying locus (Ny-Smira) to the long arm of potato chromosome IX. This position corresponds to the location of the Ry chc and Ny-1 genes for PVY resistance. A simple PCR marker, located 1 cM from the Ny-Smira gene, can be recommended for selection of PVY-resistant progeny of cv. Sárpo Mira. 相似文献
12.
Shaonpius Mondal Erik J. Wenninger Pamela J.S. Hutchinson Jonathan L. Whitworth Deepak Shrestha Sanford D. Eigenbrode Nilsa A. Bosque‐Pérez 《Entomologia Experimentalis et Applicata》2016,158(3):258-268
Potato virus Y (PVY) strains are transmitted by different aphid species in a non‐persistent, non‐circulative manner. Green peach aphid (GPA), Myzus persicae Sulzer, is the most efficient vector in laboratory studies, but potato aphid (PA), Macrosiphum euphorbiae Thomas (both Hemiptera: Aphididae, Macrosiphini), and bird cherry‐oat aphid (BCOA), Rhopalosiphum padi L. (Hemiptera: Aphididae, Aphidini), also contribute to PVY transmission. Studies were conducted with GPA, PA, and BCOA to assess PVY transmission efficiency for various isolates of the same strain. Treatments included three PVY strains (PVYO, PVYN:O, PVYNTN) and two isolates of each strain (Oz and NY090031 for PVYO; Alt and NY090004 for PVYN:O; N4 and NY090029 for PVYNTN), using each of three aphid species as well as a sham inoculation. Virus‐free tissue‐cultured plantlets of potato cv. Russet Burbank were used as virus source and recipient plants. Five weeks post inoculation, recipient plants were tested with quantitative DAS‐ELISA to assess infection percentage and virus titer. ELISA‐positive recipient plants were assayed with RT‐PCR to confirm presence of the expected strains. Transmission efficiency (percentage infection of plants) was highest for GPA, intermediate for BCOA, and lowest for PA. For all aphid species, transmission efficiency did not differ significantly between isolates within each strain. No correlations were found among source plant titer, infection percentage, and recipient plant titer. For both GPA and BCOA, isolates of PVYNTN were transmitted with greatest efficiency followed by isolates of PVYO and PVYN:O, which might help explain the increasing prevalence of necrotic strains in potato‐growing regions. Bird cherry‐oat aphid transmitted PVY with higher efficiency than previously reported, suggesting that this species is more important to PVY epidemiology than has been considered. 相似文献
13.
14.
Szajko K Chrzanowska M Witek K Strzelczyk-Zyta D Zagórska H Gebhardt C Hennig J Marczewski W 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2008,116(2):297-303
Hypersensitive resistance (HR) is an efficient defense strategy in plants that restricts pathogen growth and can be activated
during host as well as non-host interactions. HR involves programmed cell death and manifests itself in tissue collapse at
the site of pathogen attack. A novel hypersensitivity gene, Ny-1, for resistance to Potato virus Y (PVY) was revealed in potato cultivar Rywal. This is the first gene that confers HR in potato plants both to common and necrotic
strains of PVY. The locus Ny-1 mapped on the short arm of potato chromosome IX, where various resistance genes are clustered in Solanaceous genomes. Expression
of HR was temperature-dependent in cv. Rywal. Strains PVYO and PVYN, including subgroups PVYNW and PVYNTN, were effectively localized when plants were grown at 20°C. At 28°C, plants were systemically infected but no symptoms were
observed. In field trials, PVY was restricted to the inoculated leaves and PVY-free tubers were produced. Therefore, the gene
Ny-1 can be useful for potato breeding as an alternative donor of PVY resistance, because it is efficacious in practice-like resistance
conferred by Ry genes. 相似文献
15.
María del Rosario Herrera Laura Jara Vidalon Juan D. Montenegro Cinzia Riccio Frank Guzman Ida Bartolini Marc Ghislain 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2018,131(9):1925-1938
Key message
We have elucidated the Andigena origin of the potato Ryadg gene on chromosome XI of CIP breeding lines and developed two marker assays to facilitate its introgression in potato by marker-assisted selection.Abstract
Potato virus Y (PVY) is causing yield and quality losses forcing farmers to renew periodically their seeds from clean stocks. Two loci for extreme resistance to PVY, one on chromosome XI and the other on XII, have been identified and used in breeding. The latter corresponds to a well-known source of resistance (Solanum stoloniferum), whereas the one on chromosome XI was reported from S. stoloniferum and S. tuberosum group Andigena as well. To elucidate its taxonomic origin in our breeding lines, we analyzed the nucleotide sequences of tightly linked markers (M45, M6) and screened 251 landraces of S. tuberosum group Andigena for the presence of this gene. Our results indicate that the PVY resistance allele on chromosome XI in our breeding lines originated from S. tuberosum group Andigena. We have developed two marker assays to accelerate the introgression of Ryadg gene into breeding lines by marker-assisted selection (MAS). First, we have multiplexed RYSC3, M6 and M45 DNA markers flanking the Ryadg gene and validated it on potato varieties with known presence/absence of the Ryadg gene and a progeny of 6,521 individuals. Secondly, we developed an allele-dosage assay particularly useful to identify multiplex Ryadg progenitors. The assay based on high-resolution melting analysis at the M6 marker confirmed Ryadg plex level as nulliplex, simplex and duplex progenitors and few triplex progenies. These marker assays have been validated and can be used to facilitate MAS in potato breeding.16.
Somatic Hybrids Between Potato and S. berthaultii Show Partial Resistance to Soil‐Borne Fungi and Potato Virus Y
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Oumèma Nouri‐Ellouz Mohamed Ali Triki Rania Jbir‐Koubaa Assawer Louhichi Safa Charfeddine Noureddine Drira Radhia Gargouri‐Bouzid 《Journal of Phytopathology》2016,164(7-8):485-496
Three tetraploid somatic hybrid lines produced by protoplast fusion between a dihaploid potato, Solanum tuberosum, cultivar BF15 and the wild potato species Solanum berthaultii were evaluated here for their response to different soil‐borne pathogens, that is Fusarium solani, Pythium aphanidermatum and Rhizoctonia solani as well as to infection by potato virus Y (PVY). Both hybrid and BF15 plants grown in vitro were inoculated with the tested pathogen strains, that is R. solani, P. aphanidermatum, or F. solani. The growth level and disease severity index of these plants were compared to the susceptible commercial cultivar Spunta. A better growth of inoculated hybrid plants and restricted disease symptoms were observed in comparison with the commercial plants. Under glasshouse conditions and after inoculation with R. solani and P. aphanidermatum, improved resistance of the hybrid plants to these pathogens was confirmed. Indeed, these plants showed no significant damage following inoculation and a better development in R. solani‐infected plants. The susceptibility of the hybrid tubers to R. solani, P. aphanidermatum, and to F. solani infection was also determined. A significant reduction of tissue colonisation was observed in all the hybrid lines compared to the cultivated cultivars. The STBc and STBd hybrids also showed improved resistance to the PVY ordinary strain (PVYo) under glasshouse conditions. 相似文献
17.
Expression of genes for resistance to potato virus Y in potato plants and protoplasts 总被引:1,自引:0,他引:1
Plants of several potato clones with major gene resistance to potato virus Y (PVY) developed necrotic local lesions and systemic necrosis after manual inoculation with common (PVYo) or veinal necrosis (PVYN) strains of the virus. The clones reacted similarly, although their resistance genes are thought to be derived from four different wild species of Solarium. Mesophyll protoplasts from each clone became infected when inoculated with RNA of PVYo by the polyethylene glycol method. The proportion of protoplasts infected, assessed by staining with fluorescent antibody to virus particles, was similar to that of protoplasts of susceptible potato cultivars. In contrast, plants of potato cultivars Corine and Pirola, which possess gene Ry from S. stoloniferum, developed few or no symptoms when manually inoculated or grafted with PVYo. Moreover, only very few protoplasts of these cultivars produced virus particle antigen after inoculation with PVYo RNA. The extreme resistance to PVY of cvs Corine and Pirola was therefore expressed by inoculated protoplasts whereas the resistance of the necrotic-reacting potato clones was not. 相似文献
18.
Xiaohui Zhan Fengjuan Zhang Ziyang Zhong Ruhao Chen Yong Wang Ling Chang Ralph Bock Bihua Nie Jiang Zhang 《Plant biotechnology journal》2019,17(9):1814-1822
CRISPR/Cas systems provide bacteria and archaea with molecular immunity against invading phages and foreign plasmids. The class 2 type VI CRISPR/Cas effector Cas13a is an RNA‐targeting CRISPR effector that provides protection against RNA phages. Here we report the repurposing of CRISPR/Cas13a to protect potato plants from a eukaryotic virus, Potato virus Y (PVY). Transgenic potato lines expressing Cas13a/sgRNA (small guide RNA) constructs showed suppressed PVY accumulation and disease symptoms. The levels of viral resistance correlated with the expression levels of the Cas13a/sgRNA construct in the plants. Our data further demonstrate that appropriately designed sgRNAs can specifically interfere with multiple PVY strains, while having no effect on unrelated viruses such as PVA or Potato virus S. Our findings provide a novel and highly efficient strategy for engineering crops with resistances to viral diseases. 相似文献
19.
20.
A multiplex PCR approach to simultaneously genotype potato towards the resistance alleles Ry-f
sto
and Ns 总被引:1,自引:0,他引:1
Kamil Witek Danuta Strzelczyk-Żyta Jacek Hennig Waldemar Marczewski 《Molecular breeding : new strategies in plant improvement》2006,18(3):273-275
A simple and robust multiplex PCR approach was developed for detection of the alleles Ry-f
sto
and Ns conferring resistance of potato to Potato Virus Y (PVY) and Potato Virus S (PVS), respectively. Cleaved amplified polymorphic sequence (CAPS) markers GP122564 linked to Ry-f
sto
and SC811260 linked to Ns were amplified in one PCR reaction and identified after simultaneous digestion of the amplicons with restriction enzymes EcoRV and MboI. Effectiveness of this procedure for marker-assisted selection was confirmed in 55 potato cultivars. 相似文献