23 research outputs found
A Solanum stoloniferum eredetű burgonya Y vĂrus immunitás gĂ©n (Rysto) finomtĂ©rkĂ©pezĂ©se Ă©s lokalizálása a tetraploid burgonya genomban = Fine mapping and localisation of potato virus Y immunity gene of Solanum stoloniferum (Rysto) in tetraploid potato genome
Jelen kutatás cĂ©lja a Solanum stoloniferum vad burgonya fajbĂłl származĂł burgonya Y vĂrus extrĂ©m rezisztencia gĂ©n (Rysto) finomtĂ©rkĂ©pezĂ©se Ă©s lokalizálása volt a tetraploid burgonya genomban. Munkánk során előállĂtottunk egy 1100 F1 egyedet tartalmazĂł tĂ©rkĂ©pezĹ‘ populáciĂłt, amely hasad az Rysto gĂ©nre nĂ©zve. Minden egyes genotĂpust hagyományos rezisztencia tesztekkel fenotipizáltunk, majd kĂ©t - az Rysto gĂ©nhez kapcsolt - markerrel elvĂ©geztĂĽk genotipizálásukat is. A populáciĂł 1:1 hasadási arányt mutatott, tehát a gĂ©n szimplex állapotban van jelen a rezisztens szĂĽlĹ‘ genomjában. A populáciĂłbĂłl 88 genotĂpust vĂ©letlenszerűen kiválasztva nĂ©gy kĂĽlönbözĹ‘ technikával megkĂsĂ©reltĂĽnk a gĂ©nnel szorosan kapcsolt további markereket azonosĂtani. Kutatásaink során Intron- targeting technikával azonosĂtottunk egy Ăşj markert a burgonya kataláz gĂ©njĂ©ben, amely kapcsolt a Rysto gĂ©nnel. Mivel a kataláz gĂ©n helyzete ismert, Ăgy a marker lehetĹ‘vĂ© tette a Rysto gĂ©n helyzetĂ©nek egyĂ©rtelmű meghatározását is a burgonya XII. kromoszĂłmájára. További Ăşjabb, a gĂ©nnel szorosan kapcsolt markert nem tudtunk azonosĂtani, azonban a detektált polimorf szekvenciákbĂłl rĂ©szleges kapcsoltsági tĂ©rkĂ©pet szerkesztettĂĽnk a tĂ©rkĂ©pezĹ‘ populáciĂł kĂ©t szĂĽlĹ‘partnerĂ©ben. A tĂ©rkĂ©p a kĂ©sĹ‘bbiekben alkalmas lehet egyĂ©b, a nemesĂtĂ©s szempontjábĂłl jelentĹ‘s gĂ©nek/tulajdonságok tĂ©rkĂ©pezĂ©sĂ©re is. | The aims of present research project were fine mapping and localisation of extreme resistance gene Rysto originating from Solanum stoloniferum in the tetraploid potato genome. In the research project a mapping population segregating for Rysto gene with 1100 F1 individuals was developed. The phenotype of each individual was determined with conventional serological methods, while their genotype was determined also based on the use of two markers linked to the Rysto gene. The results revealed 1:1 segregation ratio and indicates the presence of a single dominant resistance gene in simplex state in the genome of resistant parent. As a subset population 88 F1 individuals was randomly chosen from the mapping population and four different molecular techniques were applied to identify further markers linked to Rysto gene. During the project one new marker linked to the Rysto gene was identified in the catalase gene of potato by Intron targeting method. Because the chromosomal position of catalase gene is known, it allowed the exact localisation of Rysto gene on chromosome XII of potato. We could not detect additional new markers closely linked to the gene, however by the use of polymorphic sequences were constructed a partial linkage map in both parents of mapping population. In the future these maps could be used for the mapping of other genes/characters being important for breeding purposes
A White Lady burgonyafajta - Solanum demissum eredetű - fitoftĂłra rezisztenciájának vizsgálata molekuláris markerek Ă©s kapcsoltsági tĂ©rkĂ©p segĂtsĂ©gĂ©vel = Investigation of late blight resistance of Solanum demissum origin in potato cv. White Lady with the help of molecular markers and linkage map
A pályázat cĂ©lja a saját nemesĂtĂ©sű White Lady fajta fitoftĂłra rezisztenciájának vizsgálata volt, mestersĂ©ges fertĹ‘zĂ©si Ă©s molekuláris genetikai vizsgálatokkal. A keszthelyi fajták fajták fĹ‘ rezisztenciaforrása a Solanum demissum vad faj, amely 11 kĂĽlönbözĹ‘ rezisztencia gĂ©nt hordoz (R1-R11). ElĹ‘zetes eredmĂ©nyeink alapján a White Lady szĂ©leskörű rezisztenciával rendelkezik a fitoftĂłra fertĹ‘zĂ©sĂ©vel szemben, azonban a rezisztenciáért felelĹ‘s gĂ©nek nem ismertek. A pályázat cĂ©lja volt a White Lady fajta S. demissum eredetű fitoftĂłra rezisztencia gĂ©njeinek meghatározása, a gĂ©nek tĂ©rkĂ©pezĂ©se, valamint kapcsolt markerek fejlesztĂ©se. KĂĽlönbözĹ‘ fitoftĂłra izolátumokkal valĂł mestersĂ©ges fertĹ‘zĂ©ssel megállapĂtottuk, hogy a White Lady rezisztenciájáért nagy valĂłszĂnűsĂ©ggel az R5-ös gĂ©n a felelĹ‘s. TesztkeresztezĂ©s utĂłdainak elemzĂ©sĂ©vel megállapĂtottuk, hogy a gĂ©n szimplex formában van jelen a fajtában. Molekuláris genetikai vizsgálatokkal igazoltuk, hogy a White Lady fajta az R5 gĂ©nen kĂvĂĽl az R2, R3a Ă©s R3b, korábban már klĂłnozott gĂ©neket is hordozza, mĂg R1 Ă©s R8 gĂ©neket nem. Kapcsoltsági tĂ©rkĂ©p vizsgálati eredmĂ©nyeink szerint az R5 gĂ©n valĂłszĂnűleg az V. kromoszĂłmán helyezkedik el, azonban hozzá kapcsolt markert nem sikerĂĽlt kifejlesztenĂĽnk. Az R2, R3a, R3b gĂ©nek kimutatása ugyanakkor lehetĹ‘sĂ©get ad ezen gĂ©nek marker alapĂş szelekciĂłjára, amivel lĂ©nyegesen hatĂ©konyabbá tehetĹ‘ a fitoftĂłrával szemben rezisztencia-nemesĂtĂ©s, megĹ‘rizhetĹ‘ a Központ kedvezĹ‘ nemzetközi versenyhelyzete. | The task of the project was to evaluate the late blight resistance of potato cv. White Lady by artificial infection and molecular genetic studies. The main source of late blight resistance of Keszthelys bred varieties is the wild species Solanum demissum that carries 11 different resistance genes to this pathogen (R1-11). Earlier studies showed that White Lady possess wide range of resistance to late blight, however the genes responsible for the resistance were not known. The goal of the project was to determine the S. demissum based resistance genes of White Lady, mapping of the genes and development of molecular markers. By artificial infections with different Phytophtora isolates we proved that most probably the gene R5 is responsible for the resistance in White Lady. Evaluating progenies of test crosses we proved that this gene is in simplex format in this variety. It was proved by molecular studies that above the gene R5, White Lady carry the genes of R2, R3a and R3b while not the gene R1 and R8. Based on our study with a highly saturated genetic map the R5 gene is most probably localized on chromosome V, however we were not able to develop linked markers to this gene yet. By the way the identification of the genes of R2, R3a and R3b gives an opportunity for the molecular selection of these genes in breeding lines making more effective the late blight resistance breeding of the Centre and helps to keep its international competitiveness
Application of direct PCR in rapid rDNA ITS haplotype determination of the hyperparasitic fungus Sphaeropsis visci (Botryosphaeriaceae)
Abstract Background The plant pathogenic fungus, Sphaeropsis visci a dark-spored species of Botryosphaeriaceae, which causes the leaf spot disease of the European mistletoe (Viscum album). This species seems to have potential as a tool for biological control of the hemiparasite. For the rapid detection of S. visci haplotypes we tested a direct PCR assay without prior DNA purification. This approach was based on a polymerase enzyme from the crenarchaeon Sulfolobus solfataricus engineered by fusion protein technology, which linked the polymerase domain to a sequence non-specific DNA binding protein (Sso7d). Findings Most isolates of Sphaeropsis visci grouped together in our phylogenetic analyses, indicating that isolates had a previously reported haplotype sequence, which is commonly found in the analyzed Hungarian population. This haplotype was also reported from diseased mistletoe bushes from other European countries. We further identified unique single nucleotide polymorphisms (SNPs) in the ITS region, which were specific to the only well resolved clade in the phylogenetic analysis. Conclusions The diPCR approach allowed amplification of ITS rRNA gene directly from small amounts of fungal samples without prior DNA extraction. This simple bioassay in plant disease management enables collection of genomic data from fungal plant pathogen populations.Peer reviewe
PICcalc: an online program to calculate polymorphic information content for molecular genetic studies
http://w3.georgikon.hu/pic/english/default.aspxPeer reviewe