Resistance Gene Analog Markers Research Articles

Genetic diversity and relationships among wild and cultivated Ficus carica L.: Usefulness of RGA markers

Disease resistance and the maintenance of genetic diversity in wild and cultivated populations are very important challenges to implement breeding program and markers assisted selection of Ficus carica L. facing climate change and its consequences. Resistance gene analogs (RGA) markers were used for variety discrimination and assessment of genetic structure and diversity of wild and cultivated Ficus carica L. species in Tunisia. The RGA markers were efficient and reliable markers for discriminating wild and cultivated fig. The high level of polymorphism (95.65) detected suggests the effectiveness of RGAs for both genetic fingerprinting and relationships assessment in wild and cultivated fig. The detected markers may represent candidate genes for disease resistance and could be further used to facilitate the identification of candidate genes and accelerate the genetic improvement of disease resistance in breeding programs of Ficus carica species.

Quantitative trait loci mapping of adult‐plant resistance to powdery mildew in triticale

AbstractPowdery mildew (PM) is a common disease caused by Blumeria graminis, which affects cereals and has recently adapted to triticale. Adult‐plant resistance (APR) genes provide durable protection of crops from the disease. Quantitative trait loci corresponding to the APR effects were mapped in an F2 population of “Lamberto” (susceptible) × “Moderto” (resistant). A genetic map of winter triticale was constructed based on the segregation of 863 DArT, 38 microsatellite and 10 resistance gene analogue markers. Composite interval mapping (CIM) was applied to identify three QTLs for maximum disease severity (MDS) and two for the area under disease progress curve (AUDPC) conferring resistance to the powdery mildew on chromosomes: 6A, 7A, 1B and 4R. The 39% variation in AUDPC was explained by the main QTL localised on chromosome 4R. Genes coding TRIUR3 proteins, serine/threonine protein kinase and cell wall associated kinases were localised in silico within the QTL and alternative DNA markers were proposed for flexible use in laboratories of diversified throughput.

Genetic Diversity Analysis Using Resistance Gene Analog-Based Markers to Support Morphological Characterization of Shallots

<p>Shallot (Allium cepa var. aggregatum) is one of the most important vegetable crops grown in Indonesia. The limited knowledge available on the genetic diversity and the threat of plant disease have been major problems to maintain high shallot production in Indonesia. Development of molecular markers linked to disease resistance is required for molecular breeding activity in this crop. This study aimed to assess the genetic diversity at conserved domain of resistance gene analog (RGA) in a set of 36 Indonesian shallot genotypes to complement morphological characterization. Twelve morphological and fifteen molecular markers traits were investigated in an attempt to characterize and to discriminate the Indonesian shallots genotypes. Characterization at orphological level indicated that phenotypic variance was highest for total bulb weight (TWB, cv = 99.39%) and the least for the plant height (PH, cv = 28.16%). The correlation analysis between traits showed that TWB and number of bulb (NB), TWB and bulb weight per plant (WB), NB and WB, and WB and PH were positively correlated. Molecular analysis revealed a total of 1,512 alleles with an average of 1.946 alleles per locus. The Polymorphism Information Content (PIC) values ranged from 0.253 to 0.676 and six out of 15 RGA markers were highly informative with PIC values ≥0.50. Based on cluster analysis, the 36 Indonesian shallot genotypes were clearly discriminated into six major groups. These results revealed that the RGA-based markers could support the morphological characterization in evaluating the genetic diversity of shallots. </p>

Relationship between Psidium species (Myrtaceae) by resistance gene analog markers: focus on nematode resistance.

Guava (Psidium guajava L.) crop is severely affected by the nematode Meloidogyne enterolobii. Native Psidium species have been reported as sources of resistance against this nematode. Knowledge on the molecular relationship between Psidium species based on plant resistance gene analogs (RGA) can be useful in the genetic breeding of guava for resistance to M. enterolobii. In this study, RGA markers from conserved domains, and structural features of plant R genes, were employed to characterize Psidium species and establish genetic proximity, with a focus on nematode resistance. SSR markers were also applied owing to their neutral nature, thus differing from RGA markers. For this, species reported as sources of resistance to M. enterolobii, such as P. cattleianum and P. friedrichsthalianum, as well as species occurring in the Atlantic Rainforest and susceptible genotypes, were investigated. In 10 evaluated Psidium species, high interspecific genetic variability was verified through RGA and SSR markers, with intraspecific variation in P. guajava higher with SSR, as was expected. Resistant species were clustered by RGA markers, and differential amplicons among genotypes resistant and susceptible to M. enterolobii were identified. Knowledge on the molecular relationships between Psidium species constitutes useful information for breeding of the guava tree, providing direction for hybridization and material for rootstocks. Additionally, the genetic relationship between native species, which have been little studied, and P. guajava were estimated by RGAs, which were confirmed as important markers for genetic diversity related to pathogen resistance.

Assessment of resistance gene analog, peroxidase, and WRKY genepolymorphisms in the genus Lens Miller

Resistance gene analogs (RGAs), peroxidases (POXs), and WRKY transcription factors are gene families related to stress-response pathways that regulate enhancement of stress tolerance in crop plants. In this study, the polymorphisms of these gene families were determined in the genus Lens Miller and Vicia montbretii (Fish. And Mey) Davis and Plitmann. A total of 32 genotypes of 7 lentil taxa, including 1 accession of cultivated lentil (Lens culinaris Medik. Subsp. culinaris), 11 accessions of L. Culinaris subsp. Orientalis (Boiss.) Ponert, 3 accessions of L. Culinaris subsp. Tomentosus Ladiz., 4 accessions of L. Culinaris subsp. Odemensis Ladiz., 3 accessions of L. Ervoides (Brign.) Grande, 4 accessions of L. Lamottei Czefr., 4 accessions of L. Nigricans (M.Bieb.) Godr., and 2 accessions of V. Montbretii, were used. Sixty-eight RGA, 22 POX, and 13 WRKY primers were used for molecular analysis. After using these degenerate primers in polymerase chain reaction, the highest polymorphism rate (90%) was observed in WRKY, followed by POX (89%) and RGA (79%). Lens culinaris Medik. Subsp. Culinaris had the lowest number of RGA and POX bands. Most RGA markers were obtained from L. Culinaris subsp. Tomentosus, L. Culinaris subsp. Orientalis, L. Culinaris subsp. Odemensis, L. Ervoides, and L. Nigricans. Similarly, the number of WRKY markers was the highest in L. Nigricans and L. Culinaris subsp. Orientalis. Lens nigricans had the highest number of POX markers among lentils. The highest numbers of RGA, POX, and WRKY markers in L. Nigricans and L. Culinaris subsp. Orientalis were in agreement with resistance for biotic and abiotic stresses, as these two taxa in the genus Lens Miller were resistant to many stresses. These results indicate that primers related to RGA, POX, and WRKY gene families may be considered potential markers for developing stress-related resistance genotypes.

Using RNA Sequencing and In Silico Subtraction to Identify Resistance Gene Analog Markers for Lr16 in Wheat.

Leaf rust, caused by Puccinia triticina Eriks., is one of the most widespread diseases of wheat and breeding for resistance is one of the most effective methods of control. Lr16 is a wheat leaf rust resistance gene (R-gene) that provides resistance at both the seedling and adult stages. Simple-sequence repeat (SSR) markers have been used to map Lr16 to the distal end of chromosome 2B. The objectives of this study were to use RNA sequencing (RNA-seq) and in silico subtraction to identify new R-gene analogs (RGAs) and use them as Lr16 markers. RNA was isolated from the susceptible wheat cultivar Thatcher (Tc) and the resistant Tc isolines TcLr10, TcLr16, TcLr21, and sequenced using Illumina technology. Using in silico subtraction, sequences from the resistant Tc isolines were aligned to a Tc reference expressed sequence tag (EST) set. Sequences not aligning to the Tc reference were assembled into contigs and analyzed using BLASTx to determine putative gene functions. Primer pairs were designed for 181 RGA sequences, of which, 137 amplified in at least one of the parents. A mapping population was developed with 165 F2 lines from a cross between the rust-susceptible cultivar Chinese Spring (CS) and TcLr16. Two RGA markers XTaLr16_RGA266585 and XTaLr16_RGA22128 were identified that mapped proximally 1.2 and 23.8 cM from Lr16, respectively. Three SSR markers Xwmc764, Xwmc661, and Xbarc35 mapped between these two RGA markers at distances of 5.0, 10.9, and 16.1 cM from Lr16, respectively. In silico subtraction is an effective technique for isolating RGAs linked to R-genes of interest.

Determination of the population structure of common bean (Phaseolus vulgaris L.) accessions using lipoxygenase and resistance gene analog markers

The common bean (Phaseolus vulgaris L.) is an important food legume throughout the world. Because of the conservation across different plant species, it is possible to evaluate the degree of genetic diversity in the common bean using gene-based marker techniques. The lipoxygenase (LOX) and resistance gene analog (RGA) genes play an important role in the response to biotic and abiotic stresses. Eighty-six common bean accessions were genotyped using gene-based LOX and RGA markers. The total number of polymorphic bands ranged from 193 for LOX to 17 for RGA markers. We detected considerable diversity with a mean of 8.7 alleles per primer for the LOX analysis. For the RGA markers, the number of alleles per polymorphic locus varied from 1 to 4 with an average allele number of 2.8. The genetic similarity between the accessions based on the LOX and RGA markers ranged from 0.12 to 0.55. Using STRUCTURE, 3 groups were revealed among the accessions. The results of this study should provide valuable information for future studies on the genetic diversity of common bean accessions and for association mapping studies examining the relationships between the genotypic and phenotypic traits related to the stress response.

Mining, genetic mapping and expression analysis of EST-derived resistance gene homologs (RGHs) in cotton.

BackgroundCotton is the dominant textile crop and also serves as an important oil crop. An estimated 15% economic loss associated with cotton production in China has been caused by diseases, and no resistance genes have been cloned in this crop. Molecular markers developed from resistance gene homologues (RGHs) might be tightly linked with target genes and could be used for marker-assisted selection (MAS) or gene cloning.ResultsTo genetically map expressed RGHs, 100 potential pathogenesis-related proteins (PRPs) and 215 resistance gene analogs (RGAs) were identified in the cotton expressed sequence tag database, and 347 specific primers were developed. Meanwhile, 61 cotton genome-derived RGA markers and 24 resistance gene analog polymorphism (RGAP) markers from published papers were included to view their genomic distribution. As a result, 38 EST-derived and 17 genome-derived RGH markers were added to our interspecific genetic map. These 55 markers were distributed on 18 of the 26 cotton chromosomes, with 34 markers on 6 chromosomes (Chr03, Chr04, Chr11, Chr17, Chr19 and Chr26). Homologous RGHs tended to be clustered; RGH clusters appeared on 9 chromosomes, with larger clusters on Chr03, Chr04 and Chr19, which suggests that RGH clusters are widely distributed in the cotton genome. Expression analysis showed that 19 RGHs were significantly altered after inoculation with the V991 stain of Verticillium dahliae. Comparative mapping showed that four RGH markers were linked with mapped loci for Verticillium wilt resistance.ConclusionsThe genetic mapping of RGHs confirmed their clustering in cotton genome. Expression analysis and comparative mapping suggest that EST-derived RGHs participate in cotton resistance. RGH markers are seemed to be useful tools to detected resistance loci and identify candidate resistance genes in cotton.

Identification of a candidate gene for resistance to root‐knot nematode in a wild peach and screening of its polymorphisms

AbstractRoot‐knot nematode disease, caused by Meloidogyne species, is an important soil‐borne disease of peach (Prunus persica L.) worldwide. To identify a major locus of genetic resistance to M. incognita, PkMi, in a wild peach species, we reconstructed a linkage group in a BC1 population of 187 lines using resistance gene analogue markers surrounding the PkMi locus. A resistance gene analogue marker, ppa021062m, co‐segregated with the PkMi locus and was therefore considered a strong candidate for PkMi. Phylogenetic analysis of the deduced protein sequences of ppa021062m, together with the other seven genes for nematode resistance, allowed ppa021062m to be assigned to the Toll/Interleukin1 Receptor‐Nucleotide Binding Site‐Leucine Rich Repeat class, similar to Ma in myrobalan plum (P. cerasifera). Comparative analysis of the candidate gene sequence in four genotypes that had different levels of resistance to root‐knot nematode disease showed that most non‐synonymous SNPs in the genic region were distributed in the TIR and NBS motifs. This study enhances our understanding of the genetic and molecular control of resistance to root‐knot nematode disease in peach.

Genetic characterization of diplodia resistant in somatic hybrids of Citrus unshiu and Citrus nobilis using resistance gene analogue (RGA)

Information about disease resistant in somatic hybrids is essential in breeding program. RGA markers are useful tools for identification, tagging, and mapping genes for disease resistant. About 4 RGA primers were used to analyze genetic character among 20 somatic hybrids of Citrus unshiu and Citrus nobilis that vary in their resistance to diplodia disease. S1/AS1, Pt8 and 18P33 primers didn't result expected band in amplification products. PCR amplification of RPS2 primers resulted expected bands size 660 bp in SS 1, SS 2 and SS 10 and unexpected bands size 110 bp of all samples. Expected band in SS 1 as resistance sample to diplodia was potential as resistance gene candidate. Protein sequences of expected band and unexpected band in amplification products of RPS2 primers showed amino acid similarity in 70% and there were kinase 2 and kinase 3a as homolog conserved domain.18P33 and RPS2 primers resulted polymorphic bands were selected for dendogram. Dendogram showed clustering of genetic similarity in 91% and all samples has genetic similarity in 100%. It was indicated that 18P33 and RPS2 primers could not prove distinction genetic character of diplodia resistant.

Comparative analysis of diversification and population structure of kenaf (Hibiscus cannabinus L.) and roselle (H. sabdariffa L.) using SSR and RGA (resistance gene analogue) markers

Multiple DNA marker systems and complementary analytical approaches are often useful in population genetic analysis and speciation of plants. We investigated population structure of kenaf (Hibiscus cannabinus) and roselle (H. sabdariffa) for gaining insight in evolution and geographic separation of these crop species using SSR and RGA (resistance gene analogues) markers through Bayesian clustering and principal coordinate analysis (PCoA) methods. Genotyping by 12 SSR and 16 RGA markers amplified a total of 172 loci in the study population. The RGA markers generated higher number of alleles per marker (8.2) as compared to SSR (3.4), but exhibited lower heterozygosity in the population. Genetic variance and heterozygosity in roselle population for both marker systems were lower than in kenaf. RGA markers revealed higher variation among populations. Bayesian structure as well as PCoA analysis using RGA marker revealed distinct cluster for roselle, while SSR-based classification revealed high admixture. Results indicate geographic isolation and natural selection for adaptive RGA alleles in kenaf. The Indian kenaf landraces were distinct from the exotic kenaf accessions, suggesting separate lineage formation by geographic separation. Possible origin and domestication of roselle in South India is proposed.

Application of resistance gene analog markers to analyses of genetic structure and diversity in rice

Plant disease resistance gene analog (RGA) markers were designed according to the conserved sequence of known RGAs and used to map resistance genes. We used genome-wide RGA markers for genetic analyses of structure and diversity in a global rice germplasm collection. Of the 472 RGA markers, 138 were polymorphic and these were applied to 178 entries selected from the USDA rice core collection. Results from the RGA markers were similar between two methods, UPGMA and STRUCTURE. Additionally, the results from RGA markers in our study were agreeable with those previously reported from SSR markers, including cluster of ancestral classification, genetic diversity estimates, genetic relatedness, and cluster of geographic origins. These results suggest that RGA markers are applicable for analyses of genetic structure and diversity in rice. However, unlike SSR markers, the RGA markers failed to differentiate temperate japonica, tropical japonica, and aromatic subgroups. The restricted way for developing RGA markers from the cDNA sequence might limit the polymorphism of RGA markers in the genome, thus limiting the discriminatory power in comparison with SSR markers. Genetic differentiation obtained using RGA markers may be useful for defining genetic diversity of a suite of random R genes in plants, as many studies show a differentiation of resistance to a wide array of pathogens. They could also help to characterize the genetic structure and geographic distribution in crops, including rice, wheat, barley, and banana.

Identification of RFLP and NBS/PK profiling markers for disease resistance loci in genetic maps of oats

Two of the domains most widely shared among R genes are the nucleotide binding site (NBS) and protein kinase (PK) domains. The present study describes and maps a number of new oat resistance gene analogues (RGAs) with two purposes in mind: (1) to identify genetic regions that contain R genes and (2) to determine whether RGAs can be used as molecular markers for qualitative loci and for QTLs affording resistance to Puccinia coronata. Such genes have been mapped in the diploid A. strigosa × A. wiestii (Asw map) and the hexaploid MN841801-1 × Noble-2 (MN map). Genomic and cDNA NBS-RGA probes from oat, barley and wheat were used to produce RFLPs and to obtain markers by motif-directed profiling based on the NBS (NBS profiling) and PK (PK profiling) domains. The efficiency of primers used in NBS/PK profiling to amplify RGA fragments was assessed by sequencing individual marker bands derived from genomic and cDNA fragments. The positions of 184 markers were identified in the Asw map, while those for 99 were identified in the MN map. Large numbers of NBS and PK profiling markers were found in clusters across different linkage groups, with the PK profiling markers more evenly distributed. The location of markers throughout the genetic maps and the composition of marker clusters indicate that NBS- and PK-based markers cover partly complementary regions of oat genomes. Markers of the different classes obtained were found associated with the two resistance loci, PcA and R-284B-2, mapped on Asw, and with five out of eight QTLs for partial resistance in the MN map. 53 RGA-RFLPs and 187 NBS/PK profiling markers were also mapped on the hexaploid map A. byzantina cv. Kanota × A. sativa cv. Ogle. Significant co-localization was seen between the RGA markers in the KO map and other markers closely linked to resistance loci, such as those for P. coronata and barley yellow dwarf virus (Bydv) that were previously mapped in other segregating populations.

Transferability of SSR and RGA Markers Developed in Cynodon spp. to Zoysia spp.

Bermudagrass (Cynodon spp.) and zoysiagrass (Zoysia spp.), which are both used as warm-season turfgrasses in the USA, are members of subfamily Chloridoideae and reported to be at least 55% genetically similar. To assess if molecular tools between the two species can be interchanged, 93 primer pairs corresponding to bermudagrass simple sequence repeats (SSRs) and resistance gene analogs (RGAs) were used to amplify DNA from eight zoysiagrass cultivars representing three species, Zoysia japonica, Z. matrella, and Z. japonica × Z. pacifica. From these 93 bermudagrass primer pairs, 11 produced clear amplicons in Zoysia cultivars. Alleles from the Zoysia accessions were scored, genetic similarities were calculated, and a cluster analysis was performed. Using these markers, genetic similarity estimates between zoysiagrass cultivars and bermudagrass accessions ranged from 0.16 to 0.22. The transferability of bermudagrass SSRs and RGAs to zoysiagrass cultivars was low (7–12%), but certain markers permitted detection of genetic variability among zoysiagrass cultivars.

Mapping resistance gene analogs (RGAs) in cultivated tetraploid cotton using RGA-AFLP analysis

Diseases cause significant losses in cotton production throughout the US Cotton Belt. Growing resistant cultivars can significantly improve cotton yields and effectively reduce production inputs. Disease resistance (R) genes have been isolated in numerous plant species and the R genes with domains of nucleotide binding sites (NB) and leucine rich repeats (LRR) represent the largest R gene family. Degenerate primers designed based on conserved motifs of plant disease resistance genes were used alone or in combination with AFLP primers to analyze disease resistance gene analogs (RGAs) in a recombinant inbred line (RIL) population of Pima (Gossypium barbadense) 3–79 and Upland cotton (G. hirsutum) line NM 24016. Eighty-eight polymorphic RGA markers were amplified by 8 pairs of RGA degenerate primers, while 131 polymorphic RGA-AFLP markers were produced from six pairs of RGA-AFLP primer combinations. Of the 219 polymorphic RGA and RGA-AFLP markers that were identified, 212 were assigned to 18 chromosomes and linkage groups based on existing SSR markers that are on known chromosomes. However, the RGA and RGA-AFLP markers are not evenly distributed among chromosomes in that 189 RGA and RGA-AFLP markers (88%) are assigned onto three “giant” chromosomes, i.e., C6, C12, and C15, suggesting RGA clusters in the cotton genome. Several RGA and RGA-AFLP markers were mapped to the same linkage group carrying a root-knot nematode resistance gene. The identification and mapping of RGA and RGA-AFLP markers provide a framework to facilitate marker-assisted selection of disease resistance in cotton breeding and to understand the physical relationship of cotton resistance genes.

Construction of a reference linkage map of Vitis amurensis and genetic mapping of Rpv8, a locus conferring resistance to grapevine downy mildew

Downy mildew, caused by the oomycete Plasmopara viticola, is one of the major threats to grapevine. All traditional cultivars of grapevine (Vitis vinifera) are susceptible to downy mildew, the control of which requires regular application of fungicides. In contrast, many sources of resistance to P. viticola have been described in the Vitis wild species, among which is V. amurensis Rupr. (Vitaceae), a species originating from East Asia. A genetic linkage map of V. amurensis, based on 122 simple sequence repeat and 6 resistance gene analogue markers, was established using S1 progeny. This map covers 975 cM on 19 linkage groups, which represent 82% of the physical coverage of the V. vinifera reference genetic map. To measure the general level of resistance, the sporulation of P. viticola and the necrosis produced in response to infection, five quantitative and semi-quantitative parameters were scored 6 days post-inoculation on the S1 progeny. A quantitative trait locus (QTL) analysis allowed us to identify on linkage group 14 a major QTL controlling the resistance to downy mildew found in V. amurensis, which explained up to 86.3% of the total phenotypic variance. This QTL was named 'Resistance to Plasmopara viticola 8' (Rpv8).

Suppressed recombination rate in 6VS/6AL translocation region carrying the Pm21 locus introgressed from Haynaldia villosa into hexaploid wheat

Pm21 is an effective gene for powdery mildew resistance transferred from Haynaldia villosa into common wheat cultivars. No virulence against this gene has been detected so far. A set of 42 powdery mildew isolates collected in Israel and tested in the current study also revealed no virulence against this gene. Pm21 was previously reported to be located on the short arm of 6VS/6AL translocation chromosome. We constructed a high-density genetic map of chromosome 6A, consisting of 28 PCR markers and the Pm21 gene. A comparison with previously published genetic maps of wheat chromosome 6A revealed that the recombination rate in the 6VS/6AL translocation region was poor. We assume that suppressed recombination caused by the alien H. villosa genetic material is the most reasonable explanation for the tight genetic linkage and the inadequacy between the Pm21 genetic map and the Pm21 physical map of 6A. A large number of sequence-tag sites (STS) and simple sequence repeat markers, which co-segregate with or are closely linked to the Pm21 gene, and the conversion of three resistance gene analog markers into new STS markers, provide a reliable and easy-to-use molecular tool for marker-assisted selection of Pm21 in wheat breeding programs. An additional gene, Pm31, previously reported to be derived from Triticum dicoccoides, was mapped into a similar genomic location to Pm21. Screening of the parental lines and the mapping population with Pm21 diagnostic markers clearly confirmed that the donor line of Pm31 is H. villosa and not T. dicoccoides. Therefore, we conclude that Pm21 and Pm31 refer to the same gene, derived from H. villosa, and that the designation of Pm31 as a new Pm gene was erroneous.

Caracterização genética de espécies de Passiflora por marcadores moleculares análogos a genes de resistência

O cultivo comercial do maracujá é afetado por diversos problemas fitossanitários, os quais contribuem para quebras de produção e significativa redução da vida útil dos plantios. Em algumas situações, a incidência de doenças pode inviabilizar o cultivo do maracujá. Fontes de resistência a distintas doenças têm sido identificadas em acessos de espécies de Passiflora. Neste trabalho, buscou-se avaliar a diversidade genética de acessos de oito espécies silvestres (P. setacea, P. nitida, P. serratodigitata, P. caerulea, P. gibertii, P. odontophyla, P. edulis e P. coccinea) e de um híbrido interespecífico (P. setacea x P. coccinea), utilizando marcadores moleculares análogos a genes de resistência (RGAs). Verificou-se uma grande diversidade no perfil eletroforético de RGAs nos acessos de Passiflora, permitindo a anotação de 96 amplicons polimórficos entre, pelo menos, um par de acessos. Os níveis de dissimilaridade genética (calculados exclusivamente com os marcadores RGAs) variaram entre 0,40 e 0,89 nos acessos das espécies de Passiflora avaliadas. A análise de sequência de um subgrupo destes amplicons obtidos com primers RGAs indicou que estas bandas correspondem a regiões genômicas que contêm segmentos (motivos) com identidade aos encontrados em genes de resistência previamente caracterizados em outras espécies vegetais. Desta forma, os dados indicam a existência de um repertório variado de marcadores do tipo RGA em Passiflora que podem ser potencialmente úteis em sistemas de caracterização molecular de germoplasma e em programas de melhoramento genético visando à resistência a doenças nesta cultura.

SPECIFIC MAPPING OF DISEASE RESISTANCE GENES IN TETRAPLOID CUT ROSES

Control of fungal diseases is a major constraint of cut-rose cultivation in greenhouses and in transportation around the world. Therefore, development of resistant cultivars is a promising way to reduce the use of chemicals required for controlling the diseases. Genetic analyses and breeding for resistance, however, are hampered by the high degree of heterozygosity and the polyploid nature of cultivated roses. Nucleotide-binding site (NBS) profiling of Van der Linden et al. (2004) was used as a tool enabling a more directed way of studying the genetics of resistance to pathogens responsible for diseases such as powdery mildew. NBS profiling is a multiplex screening technique, producing amplified resistance gene (R-gene) and resistance gene analogue (RGA) fragments by using degenerated primers based on the conserved motifs present in the NBS domain of resistance genes. Since RGAs are abundantly distributed and highly polymorphic within the plant genome, NBS profiling generates multiple markers of putative resistance genes. Twelve NBS degenerated primer/ restriction enzyme combinations were used to genotype the whole rose tetraploid K5 population (Yan, 2005) and its parents. To generate RGA profiles, the restriction enzymes: AluI, HaeIII, Mse and RsaI were used in combination with degenerated primers NBS1, NBS3, and NBS5a6. The profiles were dominantly scored resulting in 106 polymorphic RGA markers which segregated in a 1:1 or 3:1 ratio. Uni-and bi-parental simplex markers will be mapped on the two available AFLP/SSR K5 maps (Yan, 2005) with Joinmap 4.0. The resulting parental tetraploid maps will be used to dissect the genetic variation for resistance to powdery mildew resistance. Additional Rosaceae SSRs mentioned in the literature are currently tested on the K5 population to obtain allelic bridges between the tetraploid and diploid genetic maps in rose and related species in order to align them. These bridges will improve cross-ploidy comparisons in roses in order to strengthen cut rose breeding

UNDERSTANDING PLANT RESPONSES TO BIOTIC STRESS: ONGOING RESEARCH IN MUSA

ISHS III International Symposium on Banana: ISHS-ProMusa Symposium on Recent Advances in Banana Crop Protection for Sustainable Production and Improved Livelihoods UNDERSTANDING PLANT RESPONSES TO BIOTIC STRESS: ONGOING RESEARCH IN MUSA