Inheritance of time to flowering in cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.)

Time to flowering and maturity is an important adaptive feature in annual crops, including cowpeas (Vigna unguiculata (L.) Walp.). In West and Central Africa, photoperiod is the most important environmental variable affecting time to flowering in cowpea. The inheritance of time from sowing to flowering (f) in cowpeas was studied by crossing a photoperiod-sensitive genotype Kanannnado to a photoperiod-insensitive variety IT97D-941-1. Sufficient seed of F₁, F₂, F₃ and backcross populations were generated. The parental, F₁, F₂, F₃ and the backcross populations were screened for f under long natural days (mean daylength 13.4 h per day) in the field and the parents, F₁, F₂ and backcross populations under short day (10 h per day) conditions. The result of the screening showed that photoperiod in the field was long enough to delay flowering of photoperiod-sensitive genotypes. Photoperiod-sensitivity was found to be partially dominant to insensitivity. Frequency distribution of the trait in the various populations indicated quantitative inheritance. Additive (d) and additive × dominance (j) interactions were the most important gene actions conditioning time to flowering. A narrow sense heritability of 86% was estimated for this trait. This will result in 26 days gain in time to flowering with 5% selection intensity from the F₂ to F₃ generation. At least seven major gene pairs, with an average delay of 6 days each, were estimated to control time to flowering in this cross.     

A SNP-based association analysis for plant growth habit in worldwide cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp) Germplasm

Cowpea is a legume widely grown in Africa, North, Central and South America, and Asia. The cowpea plant growth habits consist of erect, semi-prostrate, and prostrate types. Developing a cultivar while considering plant growth habit is essential within a breeding program since the need for a particular growth habit is region-specific, and significantly depends on the end user preference. Some cowpea growers might prefer erect types over semi-prostrate and prostrate types, while others would chose prostrate types, which provide more leaves for feed supplies to livestock. However, very little is known regarding the genetics of plant growth habit in cowpea to assist plant breeders in developing suitable cowpea cultivars having the desired growth habit plus the other required features. Therefore, the objectives of this study were to conduct an association mapping for cowpea growth habit, and to identify SNP markers associated with this trait. A total of 487 cowpea genotypes were evaluated for growth habit and a total of 1031 SNPs postulated from genotyping-by-sequencing to conduct association analysis study for cowpea growth habit. Our results showed that: (1) significant differences in cowpea growth habit were identified between countries, (2) the cowpea erect-type was prevalent, and (3) ten SNP markers, C35060651_729, C35061339_799, C35062457_1855, C35072764_1384, C35080248_2355, Scaffold2771_4351, Scaffold29522_3213, Scaffold35913_2678, Scaffold53560_188, and Scaffold58098_4297, were significantly associated with cowpea growth habit. These results could be used for enhancing marker-assisted selection (MAS) in breeding programs aimed at developing cowpea cultivars having a particular growth habit.     

Genetic analysis of thrips resistance in cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> [L.] Walp.)

Flower bud thrips, Megalurothrips sjostedti is the most severe field pest of cowpea that causes massive flower abortion which eventually results to substantial yield reduction in Africa. There is paucity of information on the mode of gene actions controlling inheritance of resistance to flower bud thrips in cowpea in the literature. The objectives of study were to assess the genetic variability for thrips resistance among the cowpea germplasm, determined the mode of inheritance of genes that conferred resistance and both broad and narrow-sense heritability estimates for the inheritance of thrips resistance in cowpea. Twelve cowpea lines were used in crosses in the screen house at IITA, Ibadan. The mating was accomplished using North Carolina design II to generate 48F₁ hybrids, which were eventually evaluated with the parents. Data on number of peduncles, number of pods and number of thrips per flower were recorded and subjected to analysis of variance using random model by SAS 9.2. Significant variability was observed for most agronomic and thrip-adaptive traits among the cowpea germplasm, parental-lines and F₁ genotypes evaluated. General combining ability (GCA) and specific combining ability (SCA) mean squares were significant (P < 0.01) for number of pods per plant and other traits under the research environment. The GCA effect accounted for 68.82–80.07% of the total variation among hybrids for all traits except days to flowering; SCA explained less than 50% of the total variation. Narrow-sense heritability estimates ranged from 7.53 (days to flower) to 63.92% (number of peduncles per plant). Additive gene action largely controlled the inheritance of yield components and other traits under thrips infestation and these traits were moderately heritable.     

Genetic differentiation and diversity upon genotype and phenotype in cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> L. Walp.)

The evolution of species is complex and subtle which always associates with the genetic variation and environment adaption during active/passive spread or migration. In crops, this process is usually driven and influenced by human activities such as domestication, cultivation and immigration. One method to discover this process is to analyze the genetic diversity of those crops in different regions. This research first assessed the similarity and differentiation between genetic diversity of genotype and phenotype in 768 world-wild cowpea germplasm which were collected by USDA and US breeding programs. Totally 1048 genotyping by sequencing (GBS) derived single nucleotide polymorphisms (SNPs) and 17 agronomic traits were used to analyze the genetic diversity, distance, cluster and phylogeny. The group differentiation was analyzed based on both the genotype distances from 1048 SNP markers and the phenotypic (Mahalanobis) distance D2 from 11 traits. A consistent result of diversity in genotype (polymorphism information content, PIC) and phenotype (Shannon and Simpson index) indicated that the East Africa and South Asia sub-continents were the original and secondary regions of cowpea domestication. Both dendrograms built by genetic distance present relationship among different regions, and the Mantel coefficient showed medium correlation level (r = 0.58) between genotype and phenotype. The information of both genotypic and phenotypic differentiations may help us to understand evolution and migration of cowpea more comprehensively and also will inform breeders how to use cowpea germplasm in breeding programs.     

Genetic analysis of resistance to flower bud thrips (<Emphasis Type="Italic">Megalurothrips sjostedti</Emphasis>) in cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> [L.] Walp.)

Cowpea is an important legume in sub-Saharan Africa where its protein rich grains are consumed. Insect pests constitute a major constraint to cowpea production. Flower bud thrips (FTh) is the first major pest of cowpea at the reproductive stage and if not controlled with insecticides is capable of reducing grain yield significantly. Information on the inheritance of resistance to FTh is required to facilitate breeding of resistant cultivars. The genetics of resistance was studied in crosses of four cowpea lines. Maternal effect was implicated while frequency distributions of the F₂ and backcross generations suggest quantitative inheritance. Additive, dominance and epistatic gene effects made large contributions and since improved inbred lines are the desired product, selection should not be too severe in the early generations to allow for desirable gene recombination. This study suggested that some of the genes involved in the control of resistance to FTh are different in TVu1509 and Sanzi. Broad sense heritability ranged from 56% to 73%. Choice of maternal parent in a cross will be critical to the success of resistance breeding.     

Association mapping revealed SNP markers for adaptation to low phosphorus conditions and rock phosphate response in USDA cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.) germplasm

Cowpea (Vigna unguiculata (L.) Walp) is a legume of economic importance world-wide, especially in Western Africa, where it is an important part of the population’s diet. The rapidly increasing population growth in Africa requires substantial increase in cowpea production, which can be achieved by expanding land areas for agricultural purposes. In addition, prevalence of soil acidity in Africa constrains such an alternative since phosphorus availability, a key element for plant growth and development, is limited, thus resulting in poor cowpea production. The objectives of this study were to conduct an association analysis for adaptation to low phosphorus conditions and rock phosphate response in cowpea, and to identify SNP markers associated with these two traits. A total of 357 cowpea accessions, collected worldwide, was evaluated for phosphorus stress and response to addition of rock phosphate. Association analysis was conducted using 1018 SNPs obtained using genotyping-by-sequencing (GBS). TASSEL 5 and R were used for association mapping studies based on six different models. The results indicated that: (1) substantial variability in adaptation to low phosphorus conditions and rock phosphate response exists in the USDA cowpea accession panel; (2) ten SNP markers, C35006753_110, C35028233_482, C35072764_1384, C35084634_455, Scaffold21750_4938, Scaffold26894_5408, Scaffold41885_14420, Scaffold45170_4650, Scaffold50732_679; and Scaffold88448_741 were found to be associated with tolerance to low phosphorus conditions in cowpea, and (3) eight SNP markers, C35028233_482, C35058535_121, Scaffold26894_5408, Scaffold45170_4650, Scaffold51609_507, Scaffold53730_7339, Scaffold74389_5733, and Scaffold87916_4921 were highly associated with rock phosphate response. These SNP markers can be used in a marker-assisted breeding (MAS) program to improve cowpea tolerance to phosphorus stress.     

Efficiency of SNP and SSR-based analysis of genetic diversity,population structure,and relationships among cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.) germplasm from East Africa and IITA inbred lines

The extent of genetic diversity and relatedness of cowpea germplasm from East Africa are poorly understood. A set of 13 microsatellites (SSR) and 151 single nucleotide polymorphisms (SNPs) markers were applied to assess the levels of genetic diversity in a sample of 95 accessions of local cowpea germplasm and inbred lines of Vigna unguiculata. The average genetic diversity (D), as quantified by the expected heterozygosity, was higher for SSR loci (0.52) than for SNPs (0.34). The polymorphic information content was 0.48 for SSR and 0.28 for SNP while the fixation index was 0.095 for SSR and 0.15 for SNPs showing moderate differentiation and high gene flow among cowpea accessions from East African countries. The results of data analysis of both SSR and SNP markers showed similar clustering patterns suggesting a substantial degree of association between origin and genotype. Principal coordinate analysis (PCoA) with SSR and SNP markers showed that accessions were grouped into two and three broad groups across the first two axes, respectively. Our study found that SNP markers were more effective than SSR in determining the genetic relationship among East African local cowpea accessions and IITA inbred lines. Based on this analysis, five local cowpea accessions Tvu-13490, Tvu-6378, Tvu-13448, Tvu-16073, and 2305675 were identified to be tightly clustered sharing several common alleles with the drought tolerant variety Danila when analyzed with SSR and SNP markers. The findings will assist and contribute to future genetic diversity studies aimed at the genetic improvement of local Eastern Africa cowpea accessions for improved overall agronomic performance in general and breeding for drought tolerant in particular.     

Genome-wide SNP-based association mapping of resistance to <Emphasis Type="Italic">Phytophthora sojae</Emphasis> in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.)

Phytophthora root rot (PRR) is among the most important soybean (Glycine max (L.) Merr.) diseases worldwide, and the host displays complex genetic resistance. A genome-wide association study was performed on 337 accessions from the Yangtze-Huai soybean breeding germplasm to identify resistance regions associated with PRR resistance using 60,862 high-quality single nucleotide polymorphisms markers. Twenty-six significant SNP-trait associations were detected on chromosomes 01 using a mixed linear model with the Q matrix and K matrix as covariates. In addition, twenty-six SNPs belonged to three adjacent haplotype blocks according to a linkage disequilibrium blocks analysis, and no previous studies have reported resistance loci in this 441 kb region. The real-time RT-PCR analysis of the possible candidate genes showed that two genes (Glyma01g32800 and Glyma01g32855) are likely involved in PRR resistance. Markers associated with resistance can contribute to marker-assisted selection in breeding programs. Analyses of candidate genes can lay a foundation for exploring the mechanism of P. sojae resistance.     

Molecular diversity and association mapping of quantitative traits in Tibetan wild and worldwide originated barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) germplasm

Molecular diversity of 40 accessions of Tibetan wild barley (TB), 10 Syrian (SY), 72 North American (NA), 36 European (EU), 9 South American (SA) and 8 Australian (AU) varieties were characterized using multiple microsatellite loci. The 42 SSR primers amplified 278 alleles across the 175 barley accessions tested in the present study. The average gene diversity for the whole sample was 0.3387 whereas the mean value for the each population was as follows: TB = 0.3286, SY = 0.2474, EU = 0.299, AU = 0.2867, NA = 0.3138, SA = 0.2536. Clustering analysis based on Nei’s original genetic distance showed that the EU and NA barley populations were grouped together. The TB population was well separated from the other 5 barley populations. Associations between microsatellite markers and 14 quantitative traits were also investigated. Significant associations were found for 18 microsatellite marker loci. The number of marker loci associated with each trait ranged from one (stem diameter, filled grains per plant, grain weight per plant, length of main spike and awn length) to seven (plant height). The percentage of the total variation explained by each marker ranged from 4.59% (HVM2 associated with plant height) to 17.48% (Bmac90 associated with density of main spike). This study provides candidate markers for further QTL mapping of these traits and for marker-assisted selection.     

Fine mapping of a <Emphasis Type="Italic">Xantha</Emphasis> mutation in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The recessive mutation of the XANTHA gene (XNT) transforms seedlings and plants into a yellow color, visually distinguishable from normal (green) rice. Thus, it has been introduced into male sterile lines as a distinct marker for rapidly testing and efficiently increasing varietal purity in seed and paddy production of hybrid rice. To identify closely linked markers and eventually isolate the XNT gene, two mapping populations were developed by crossing the xantha mutant line Huangyu B (indica) with two wild type japonica varieties; a total of 1,720 mutant type F₂ individuals were analyzed for fine mapping using polymorphic InDel markers and high dense microsatellite markers. The XNT gene was mapped on chromosome 11, within in a fragment of ~100 kb, where 13 genes are annotated. The NP_001067671.1 gene within the delimited region is likely to be a candidate XNT gene, since it encodes ATP-dependent chloroplast protease ATP-binding subunit clp A. However, no sequence differences were observed between the mutant and its parent. Bioinformatics analysis demonstrated that four chlorophyll deficient mutations that were previously mapped on the same chromosome are located outside the XNT region, indicating XNT is a new gene. The results provide useful DNA markers not only for marker assisted selection of the xantha trait but also its eventual cloning.     

Identification of genetic sources with attenuated <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>-induced symptoms in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm

The whitefly-transmitted Tomato chlorosis virus (ToCV) (genus Crinivirus) is associated with yield and quality losses in field and greenhouse-grown tomatoes (Solanum lycopersicum) in South America. Therefore, the search for sources of ToCV resistance/tolerance is a major breeding priority for this region. A germplasm of 33 Solanum (Lycopersicon) accessions (comprising cultivated and wild species) was evaluated for ToCV reaction in multi-year assays conducted under natural and experimental whitefly vector exposure in Uruguay and Brazil. Reaction to ToCV was assessed employing a symptom severity scale and systemic virus infection was evaluated via RT-PCR and/or molecular hybridization assays. A subgroup of accessions was also evaluated for whitefly reaction in two free-choice bioassays carried out in Uruguay (with Trialeurodes vaporariorum) and Brazil (with Bemisia tabaci Middle-East-Asia-Minor1—MEAM1?=?biotype B). The most stable sources of ToCV tolerance were identified in Solanum habrochaites PI 127827 (mild symptoms and low viral titers) and S. lycopersicum ‘LT05’ (mild symptoms but with high viral titers). These two accessions were efficiently colonized by both whitefly species, thus excluding the potential involvement of vector-resistance mechanisms. Other promising breeding sources were Solanum peruvianum (sensu lato) ‘CGO 6711’ (mild symptoms and low virus titers), Solanum chilense LA1967 (mild symptoms, but with high levels of B. tabaci MEAM1 oviposition) and Solanum pennellii LA0716 (intermediate symptoms and low level of B. tabaci MEAM1 oviposition). Additional studies are necessary to elucidate the genetic basis of the tolerance/resistance identified in this set of Solanum (Lycopersicon) accessions.     

Heritability and gene effects for incorporating pubescence into cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L) Walp.) from <Emphasis Type="Italic">V. rhomboidea</Emphasis> Burtt. Davy

Cowpea, an important subsistence pulse crop in the tropics and subtropics is susceptible to several insect pests that seriously limit its yield potential. Pubescence (hairiness) on the parts of some of its wild relatives has been reported to reduce the damage caused by some of the insect pests. A generation mean analysis was conducted between a cowpea variety IT82D-716 and two accessions of Vigna rhomboidea to investigate the gene effects and heritability for incorporating pubescence into cultivated cowpea from V. rhomboidea. The additive-dominance model that was adopted in the analysis was observed to sufficiently explain the mode of inheritance of leaf and stem pubescence with the additive effect being more important than the dominance effect. A six-parameter model with epistatic gene interactions was adequate for explaining the inheritance of pod pubescence. Heritability estimates, in the narrow sense were high for pubescence density and pubescence length. Inheritance of pubescence in crosses between cowpea and V. rhomboidea was governed by one and two genes. Significant and higher additive gene effects and high-heritability suggest that backcross selection schemes should be responsive in the development of pubescent cowpea lines.     

Phenotypic diversity in cold-tolerant peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.) germplasm

Tolerance to low temperature is an important prerequisite for optimal performance of peanut (Arachis hypogaea L.) in a number of temperate peanut-growing environments. One hundred fifty-eight peanut accessions belonging to five botanical types, known to be tolerant to low temperature (12°C) at germination, were evaluated for phenotypic diversity for 15 morphological traits in the 2001 rainy season and for 15 agronomic and two seed quality traits in the 2001 rainy and 2001/2002 post-rainy seasons. Analysis of data, using the residual maximum-likelihood approach indicated that variance components due to genotypes were significant for all traits in the rainy and for all but two traits in the post-rainy season. Clustering based on scores of nine principle components delineated four clusters. The cold-tolerant genotypes and the standard control cultivars in the four clusters differed in mean, variance, and range both during rainy and post-rainy seasons for a range of agronomic traits, indicating the diversity among the clusters. The cold-tolerant accessions were superior to control cultivars for several agronomic traits compared with their respective controls in both the rainy and post-rainy seasons, so their use in breeding should result in genetically diverse cold-tolerant high-yielding peanut cultivars.     

Successful induction of trigenomic hexaploid <Emphasis Type="Italic">Brassica</Emphasis> from a triploid hybrid of <Emphasis Type="Italic">B.</Emphasis><Emphasis Type="Italic">napus</Emphasis> L. and <Emphasis Type="Italic">B. nigra</Emphasis> (L.) Koch

A triploid hybrid with an ABC genome constitution, produced from an interspecific cross between Brassica napus (AACC genome) and B. nigra (BB genome), was used as source material for chromosome doubling. Two approaches were undertaken for the production of hexaploids: firstly, by self-pollination and open-pollination of the triploid hybrid; and secondly, by application of colchicine to axillary meristems of triploid plants. Sixteen seeds were harvested from triploid plants and two seedlings were confirmed to be hexaploids with 54 chromosomes. Pollen viability increased from 13% in triploids to a maximum of 49% in hexaploids. Petal length increased from 1.3 cm (triploid) to 1.9 cm and 1.8 cm in the two hexaploids and longest stamen length increased from 0.9 cm (triploid) to 1.1 cm in the hexaploids. Pollen grains were longer in hexaploids (43.7 and 46.3 μm) compared to the triploid (25.4 μm). A few aneuploid offsprings were also observed, with chromosome number ranging from 34 to 48. This study shows that trigenomic hexaploids can be produced in Brassica through interspecific hybridisation of B. napus and B. nigra followed by colchicine treatment.     

Genetic effects of introgression genomic components from Sea Island cotton (<Emphasis Type="Italic">Gossypium barbadense</Emphasis> L.) on fiber related traits in upland cotton (<Emphasis Type="Italic">G</Emphasis>. <Emphasis Type="Italic">hirsutum</Emphasis> L.)

The germplasm with exotic genomic components especially from Sea Island cotton (Gossypium barbadense L. Gb) is the dominant genetic resources to enhance fiber quality of upland cotton (G. hirsutum L., Gh). Due to low efficiency of phenotypic evaluation and selection on fiber quality, genetic dissection of favorable alleles using molecular markers is essential. Genetic dissection on putative Gb introgressions related to fiber traits were conducted by SSR markers with mapping populations derived from a cross between Luyuan343 (LY343), a superior fiber quality introgression line (IL) with genomic components from Gb, and an elite Upland cotton cv. Lumianyan#22 (LMY22). Among 82 polymorphic loci screened out from 4050 SSRs, 42 were identified as putative introgression alleles. A total of 29 fiber-related QTLs (23 for fiber quality and six for lint percentage) were detected and most of which clustered on the putative Gb introgression chromosomal segments of Chr.2, Chr.16, Chr.23 and Chr.25. As expected, a majority of favorable alleles of fiber quality QTLs (12/17, not considering the QTLs for fiber fineness) came from the IL parent and most of which (11/12) were conferred by the introgression genomic components while three of the six (3/6) favorable alleles for lint percentage came from the Gh parent. Validation of these QTLs using an F₈ breeding population from the same cross made previously indicated that 13 out of 29 QTLs showed considerable stability. The results suggest that fiber quality improvement using the introgression components could be facilitated by marker-assisted selection in cotton breeding program.     

Genetic analysis of tropical quality protein maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm

Maize (Zea mays L.) is an important source of carbohydrates and protein in the diet in sub-Saharan Africa. The objectives of this study were to (i) estimate general (GCA) and specific combining abilities (SCA) of 13 new quality protein maize (QPM) lines in a diallel under stress and non-stress conditions, (ii) compare observed and predicted performance of QPM hybrids, (iii) characterize genetic diversity among the 13 QPM lines using single nucleotide polymorphism (SNP) markers and assess the relationship between genetic distance and hybrid performance, and (iv) assess diversity and population structure in 116 new QPM inbred lines as compared to eight older tropical QPM lines and 15 non-QPM lines. The GCA and SCA effects were significant for most traits under optimal conditions, indicating that both additive and non-additive genetic effects were important for inheritance of the traits. Additive genetic effects appeared to govern inheritance of most traits under optimal conditions and across environments. Non-additive genetic effects were more important for inheritance of grain yield but additive effects controlled most agronomic traits under drought stress conditions. Inbred lines CKL08056, CKL07292, and CKL07001 had desirable GCA effects for grain yield across drought stress and non-stress conditions. Prediction efficiency for grain yield was highest under optimal conditions. The classification of 139 inbred lines with 95 SNPs generated six clusters, four of which contained 10 or fewer lines, and 16 lines of mixed co-ancestry. There was good agreement between Neighbor Joining dendrogram and Structure classification. The QPM lines used in the diallel were nearly uniformly spread throughout the dendrogram. There was no relationship between genetic distance and grain yield in either the optimal or stressed environments in this study. The genetic diversity in mid-altitude maize germplasm is ample, and the addition of the QPM germplasm did not increase it measurably.     

Search in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm for sources of broad-spectrum resistance to four <Emphasis Type="Italic">Tospovirus</Emphasis> species

The genus Tospovirus was considered as monotypic with Tomato spotted wilt virus (TSWV) being the only assigned species. However, extensive studies with worldwide isolates revealed that this genus comprises a number of species with distinct virulence profiles. The Neotropical South America is one center of Tospovirus diversity with many endemic species. Groundnut ringspot virus (GRSV), TSWV, Tomato chlorotic spot virus (TCSV), and Chrysanthemum stem necrosis virus (CSNV) are the predominant tomato-infecting species in Brazil. Sources of resistance were found in Solanum (section Lycopersicon) mainly against TSWV isolates from distinct continents, but there is an overall lack of information about resistance to other viral species. One-hundred and five Solanum (section Lycopersicon: Solanaceae) accessions were initially evaluated for their reaction against a GRSV isolate by analysis of symptom expression and systemic virus accumulation using DAS-ELISA. A subgroup comprising the most resistant accessions was re-evaluated in a second assay with TSWV, TCSV, and GRSV isolates and in a third assay with a CSNV isolate. Seven S. peruvianum accessions displayed a broad-spectrum resistance to all viral species with all plants being free of symptoms and systemic infection. Sources of resistance were also found in tomato cultivars with the Sw-5 gene and also in accessions of S. pimpinellifolium, S. chilense, S. arcanum, S. habrochaites, S. corneliomuelleri, and S. lycopersicum. The introgression/incorporation of these genetic factors into cultivated tomato varieties might allow the development of genetic materials with broad-spectrum resistance, as well as with improved levels of phenotypic expression.     

Molecular mapping of <Emphasis Type="Italic">Pc68</Emphasis>, a crown rust resistance gene in <Emphasis Type="Italic">Avena</Emphasis><Emphasis Type="Italic">sativa</Emphasis>

Crown rust, which is caused by Puccinia coronata f. sp. avenae, P. Syd. & Syd., is the most destructive disease of cultivated oats (Avena sativa L.) throughout the world. Resistance to the disease that is based on a single gene is often short-lived because of the extremely great genetic diversity of P. coronata, which suggests that there is a need to develop oat cultivars with several resistance genes. This study aimed to identify amplified fragment length polymorphism AFLP markers that are linked to the major resistance gene, Pc68, and to amplify the F₆ genetic map from Pc68/5*Starter × UFRGS8. Seventy-eight markers with normal segregation were discovered and distributed in 12 linkage groups. The map covered 409.4 cM of the Avena sativa genome. Two AFLP markers were linked in repulsion to Pc68: U8PM22 and U8PM25, which flank the gene at 18.60 and 18.83 centiMorgans (cM), respectively. The marker U8PM25 is located in the linkage group 4_12 in the Kanota × Ogle reference oat population. These markers should be useful for transferring Pc68 to genotypes with good agronomic characteristics and for pyramiding crown rust resistance genes.     

<Emphasis Type="Italic">Agrobacterium</Emphasis> mediated transformation of indica rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) for insect resistance

The rice leaffolder (RLF), Cnaphalocrocis medinalis is an important pest of rice that causes severe damage in many areas of the world. The plants were transformed with fully modified (plant codon optimized) synthetic Cry1C coding sequences as well as with the hpt and gus genes, coding for hygromycin phosphotransferase and β-glucuronidase, respectively. Cry1C sequences placed under the control of doubled 35S promoter plus the AMV leader sequence, and hpt and gus genes driven by cauliflower mosaic virus 35S promoter, were used in this study. Embryogenic calli after cocultivation with Agrobacterium were selected on the medium containing hygromycin B. A total of 67 hygromycin-resistant plants were regenerated. PCR and Southern blot analyses of primary transformants revealed the stable integration of Cry1C coding sequences into the rice genome with predominant single copy integration. R₁ progeny plants disclosed a monogenic pattern (3:1) of transgene segregation as confirmed by molecular analyses. These transgenic lines were highly resistant to rice leaffolder (RLF), Cnaphalocrocis medinalis as revealed by insect bioassay.     

Identification of quantitative trait loci for bruchid (<Emphasis Type="Italic">Callosobruchus maculatus</Emphasis>) resistance in black gram [<Emphasis Type="Italic">Vigna mungo</Emphasis> (L.) Hepper]

An inter-subspecific mapping population was generated by crossing V. mungo var. mungo (cv. TU 94-2, bruchid susceptible) and V. mungo var. silvestris (bruchid resistant). About 37.8% of the bruchid completed their lifecycle on seeds of V. mungo var. silvestris compared with 100% on the susceptible variety TU 94-2. The total developmental period of C. maculatus on Vigna mungo var. silvestris was considerably extended (88 days as compared with 34 days on TU 94-2). A genetic linkage map constructed using recombinant inbred lines (RILs) in F₉ generation with 428 markers [86 random amplified polymorphic DNA (RAPD), 47 simple sequence repeat (SSR), 41 inter-SSR (ISSR), 254 amplified fragment length polymorphism (AFLP)] was used for QTL detection. One hundred four individuals were used for detection of QTLs associated with bruchid resistance. The RILs exhibited a high level of variation in percentage adult emergence (0–100%) and developmental period (0–105 days). Two QTLs, Cmrae1.1 and Cmrae1.2, were identified for percentage adult emergence, on linkage group (LG) 3 and 4, respectively. For developmental period, six QTLs were identified, with two QTLs (Cmrdp1.1 and Cmrdp1.2) on LG 1, three QTLs (Cmrdp1.3, Cmrdp1.4, and Cmrdp1.5) on LG 2, and one QTL (Cmrdp1.6) on LG 10.