Browsing by Subject "RAPD"

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Diversity in the tropical multipurpose shrub legumes Cratylia argentea (Desv.) O. Kuntze and Flemingia macrophylla (Willd.) Merrill.
(2006) Andersson, Meike S.; Schultze-Kraft, Rainer
Cratylia argentea (Desv.) O. Kuntze and Flemingia macrophylla (Willd.) Merrill are promising tropical multipurpose shrub legumes. Both are drought-tolerant, well adapted to low-fertility, acid soils, and especially suited for low-input smallholder production systems in the sub-humid and humid tropics. They can be used e.g. as dry season forage supplementation, live soil cover or mulch, erosion barrier hedges, and shade-providing shrubs in young coffee and cocoa plantations. Germplasm collections were assembled from the wild-legume flora in Brazil (C. argentea) and Southeast Asia (F. macrophylla), but research and development are so far based on only a few accessions. Knowledge about the extent of genetic diversity within these collections is very limited. In addition, the potential utilization of F. macrophylla is so far limited by poor forage quality and acceptability of the few evaluated accessions. The objective of the present study, conducted in a research cooperation with the International Centre for Tropical Agriculture (CIAT), Cali, Colombia, was to assess the diversity in the germplasm collections of C. argentea (38 accessions) and F. macrophylla (69 accessions) in terms of morphological and phenological traits, agronomic and forage quality traits, and molecular markers, and to identify superior genotyes. Based on these different characterization approaches, the objective was furthermore to establish core collections for F. macrophylla, and to compare and validate the different strategies, giving particular consideration to their practical implications (time and cost efficiency) for the application to small collections of perennial wild tropical legumes. Cratylia argentea High diversity in terms of phenological and agronomic as well as forage quality traits was detected in the collection, with scope for plant improvement in terms of higher dry season DM production. Accessions CIAT 18674 and 22406 were identified as promising for further evaluation since they were similar to the commercial cultivar "Veraniega" in terms of forage quality, and superior in terms of DM production, particularly in the dry season. Molecular marker analysis with RAPDs showed that the genetic diversity in the collection was relatively low and fairly homogeneously distributed. Accessions CIAT 22373, 22378, 22380, 22381 and 22411 were identified as possible duplicates. Flemingia macrophylla High diversity in terms of morphological and agronomic as well as forage quality traits was detected among the 69 accessions. The identification of four morphotypes in the collection probably has taxonomic implications. Scope for plant improvement was identified with respect to forage quality - one of the species´ main limitations. Accessions CIAT 18437, 21083 and 21090 had similar DM production and higher digestibility than the control accession, and were virtually free of extractable condensed tannins. Problems with low palatability and low seed production of these promising accessions need to be further studied. Genetic diversity in F. macrophylla was higher than in C. argentea, and corresponded closely to the four morphotypes revealed by conventional characterization. Various duplicate accessions were identified, and evidence was provided that the non-Asian accessions are not native to their collection site regions, but rather introduced from Southeast Asia. The results have direct applications for plant improvement of these promising multipurpose legumes. The superior genotypes selected in this study will be used in work with farmers in CIAT-research sites in Central America and distributed to partners. It must be recognized, however, that the diversity assessed is influenced by the climatic and edaphic conditions at the site where the studies were conducted. Therefore, multilocational trials should be considered with a selected subset (including the promising accessions) of C. argentea and F. macrophylla i) to assess the extent of genotype x environment interaction, and ii) to identify genotypes with consistently high performance in a range of distinct environments. Research on the reproduction system of both species is urgently required to determine the potential extent and impact of outcrossing. Beyond the immediate application of these species for farmer utilization, the results of the use and comparison of different approaches to assess diversity and to establish core collections can help to improve germplasm management and characterization of wild tropical legume species in general. Random sampling has been identified as a valuable and resource-efficient strategy for the creation of core collections when no additional information about accessions is available, and in the absence of adequate funds. The validation of the findings of this study with a broader range of perennial tropical wild legumes is necessary to assess their applicability to other species.
Molecular and phenotypic analyses of pathogenicity, aggressiveness, mycotoxin production, and colonization in the wheat-Gibberella zeae pathosystem
(2004) Cumagun, Christian Joseph R.; Miedaner, Thomas
Fusarium head blight (FHB), caused by Gibberella zeae (Schwein.) Petch (anamorph: Fusarium graminearum Schwabe), is one of the principal diseases responsible for extensive damage in wheat fields and contamination of grain with the mycotoxins deoxynivalenol (DON) and nivalenol (NIV), rendering the harvest unsafe for human and animal consumption. Control of FHB is difficult because of the complex nature of host-pathogen-environment interaction and the nonavailability of highly effective fungicides. Agronomic practices and resistance breeding, therefore, offer the best strategies for disease management. Mapping by molecular markers provides an accurate approach for genetic analyses of simple and complex traits particularly pathogenicity, aggressiveness, and mycotoxin production. Pathogenicity, as defined here, is the ability to cause disease whereas aggressiveness is the quantity of disease induced by a pathogenic isolate on a susceptible host in which isolates do not interact differentially with host cultivars. The project aims to (1) map pathogenicity and aggressiveness of G. zeae based on a published genetic map (2) estimate genetic diversity of four parent isolates by PCR-based markers (3) examine the inheritance of pathogenicity, aggressiveness, mycotoxin type (DON/NIV), and DON production on a phenotypic basis, (4) analyse genetic covariation among aggressiveness, DON, and fungal colonization, (5) and compare aggressiveness of 42 isolates in greenhouse and field environments. Two crosses of G. zeae using nit (nitrate nonutilizing) marker technique were performed: (1) pathogenic DON-producing Z-3639 (Kansas, USA) x nonpathogenic NIV-producing R-5470 (Japan) belonging to lineage 7 and 6, respectively, and (2) DON-producing FG24 (Hungary) x FG3211 (Germany), both aggressive lineage 7 isolates. For the first cross, 99 progeny segregated in a consistent 61:38 for pathogenicity: nonpathogenicity in a two-year greenhouse experiment. Among the 61 pathogenic progeny, disease severity, measured as percentage infected spikelets, varied significantly (P = 0.01). Heritability for aggressiveness was high. Pathogenicity locus was mapped on linkage group IV near loci PIG1 (red pigment production), TOX1 (trichothecene toxin amount), and PER1 (perithecial production) explaining 60%, 43%, and 51% of the phenotypic variation, respectively. Two large aggressiveness QTLs were mapped on linkage group I linked to the locus TRI5 (trichodiene synthase in the trichothecene gene cluster) and an amplified fragment length polymorphism (AFLP) marker (EAAMTG0655K), explaining 51% and 29% of the observed phenotypic variation, respectively. These unlinked loci suggest that genetic basis between pathogenicity and aggressiveness were different. TRI5 is located in the same gene cluster as a previously identified gene known as TRI13, which determines whether DON or NIV will be produced. DON-producing progeny were, on average, twice as aggressive as were those producing NIV. Loci were only detected in the two linkage groups mentioned from the nine linkage groups present in the map. For the second cross FG24 x FG3211 with 153 progeny, head blight rating and relative plot yield were used as aggressiveness traits. DON production was measured by a commercial kit enzyme immunoassay. These three traits were quantitatively inherited among 153 progeny across three environments. Repeatabilities within each environment were medium to high but heritabilities across environments were medium only due to high progeny-environment interaction. DON was a less environmentally stable trait than aggressiveness. Transgressive segregants were detected frequently. This implies that even a cross within a lineage could lead to an increase in aggressiveness. Mapping of this cross was not initiated because the parents were not polymorphic enough to construct a genetic map. Instead, the parents were analysed for polymorphism in comparison to the parents of the first cross using 31 AFLP primer combinations and 56 random amplified polymorphic DNA (RAPD) primers. Polymorphism between Z-3639 and R-5470 was about three to four times higher than between FG24 and FG3211. Cluster analysis revealed that R-5470 was genetically separated from the other three parents, thus confirming the lineage assignments. Among preselected 50 progeny from the same field experiments that showed normal distribution for aggressiveness - head blight rating, fungal colonization, and DON production were correlated (r = 0.7, P = 0.01). Fungal colonization measured as Fusarium exoantigen (ExAg) content using enzyme-linked immunosorbent assay (ELISA) varied also quantitatively, but heritability was lower due to high progeny-environment interaction and error. Strong correlations among all traits indicate control by similar genes or gene complexes. No significant variation was observed for DON/ExAg ratio. Aggressiveness traits and DON production were more environmentally stable compared to Fusarium ExAg content. Our findings imply that aggressiveness may have other components apart from mycotoxin production. Genotypic variation for aggressiveness among the 42 progeny in one greenhouse and three field environments was significant and their correlation was moderate (r = 0.7, P = 0.01). High heritability in both environments again indicates that aggressiveness was a relatively stable trait, although methods of inoculation differed, i.e., injection for greenhouse and spraying for field experiments. Greenhouse aggressiveness could predict aggressiveness in the field, and thereby should reduce costs for resistance and phytopathological studies. In conclusion, we consider G. zeae as medium-risk pathogen with the potential to evolve to a higher level of aggressiveness due to sexual recombination. Erosion of quantitative resistance in FHB cannot be ignored, especially if host resistances with oligogenic inheritance, e.g. Sumai 3 from China, are used on a large acreage. Consequently, the rather simple inheritance of pathogenicity and aggressiveness in G. zeae could lead to a gradual increase of aggressiveness. These results should enhance efforts of plant breeders to use several, genetic distinct sources of resistance in order to avoid possible FHB outbreaks in the future.