Mapping of QTLs detected in a <Emphasis Type="Italic">Brassica napus</Emphasis> DH population for resistance to <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> in multiple environments

Sclerotinia stem rot, caused by the fungus Sclerotinia sclerotiorum, is one of the most devastating diseases of rapeseed (Brassica napus L.) in China. The two major factors limiting the development of disease resistance are (1) the absence of accessions with complete resistance and (2) the lack of a single method that can be widely applied to assess tolerance—even though accessions with differential tolerance to S. sclerotiorum have been identified in China. In the study reported here, we have used one doubled haploid (DH) population consisting of 72 lines, which was derived from the F₁ generation of a cross between a partially resistant line (DH821) and a susceptible line (DHBao604), to identify quantitative trait loci (QTLs) involved in the resistance to S. sclerotiorum. Three inoculation methods, namely, mycelial toothpick inoculation (MTI), mycelial plug inoculation (MPI), and infected petal inoculation (IPI), were used to assess resistance at the adult plant stage. A genetic linkage map with 20 linkage groups covering 1746.5 cM, with an average space of 6.93 cM, was constructed using a total of 252 molecular markers, including 91 simple sequence repeats, 72 randomly amplified polymorphic DNA, 86 sequence-related amplified polymorphisms, two restriction fragment length polymorphisms, and one expressed sequence tag. Composite interval mapping identified ten, one and ten QTLs using MTI, MPI and IPI methods, respectively, at a LOD > 2.5. One QTL was detected in linkage group N12 by MTI in 2004 and 2005 and by IPI in 2005. Another QTL was detected in linkage group N3 and N4 by MPI in 2006 and 2007. There was one common QTL detected by MTI in 2005 and by MPI in 2006. These results provide information on the genetic control of resistance to S. sclerotiorum in oilseed rape.     

Papaya ringspot virus resistance in <Emphasis Type="Italic">Carica papaya</Emphasis> via introgression from <Emphasis Type="Italic">Vasconcellea quercifolia</Emphasis>

We report the first successful production of PRSV-P resistant backcross (BC) papaya plants following intergeneric hybridisation between C. papaya and a Vasconcellea species after 50 years of reports on unsuccessful attempts. This follows our previous reports of PRSV-P resistant F₁ hybrids developed by intergeneric hybridisation between C. papaya and V. quercifolia. One PRSV-P resistant BC 1 (BC₁) plant was produced after 114,839 seeds were dissected from 940 fruits. The seeds yielded 1,011 embryos and 733 germinated in vitro from which 700 developed into plantlets that were screened in a glasshouse and in the field under high disease pressure and exposure to inoculation by viruliferous aphids. From the PRSV-P resistant backcross 1 (BC₁) male plant, 1465 plants [137 BC₂, 546 SbC₂ (BC2 sib-crosses), 147 BC₃, 379 SbC₃ and 256 BC₄] were grown from seed and inoculated with PRSV-P and virus resistant BC₃ and BC₄ plants were selected from these generations. Presence or absence of virus was confirmed by ELISA serological tests. BC plants generally developed mild symptoms of PRSV-P after periods ranging from 5 to 18 months in the field but many showed the ability to produce new growth free of symptoms. All control plants developed severe symptoms after 3 months in the field. Some BC₃ and BC₄ plants were free from viral infection after 18 months in the field. Subsequently they developed very mild symptoms on their leaves and a few ringspots on their fruit. They continued to grow vigorously and produce fruit for 3 years under high disease pressure provided by the infected controls and other susceptible plants. Good quality marketable fruit were produced on these plants. Application of these results should lead to restoration of the papaya industry in virus-infested regions of the Philippines and worldwide.     

Genetic analysis reveals a dominant <Emphasis Type="Italic">S</Emphasis> locus and an <Emphasis Type="Italic">S</Emphasis> suppressor locus in natural self-compatible <Emphasis Type="Italic">Brassica napus</Emphasis>

A self-incompatible (SI) line, S-1300, and its maintainer 97-wen135, a self-compatible (SC) line, were used to study the inheritance of maintenance for self-incompatibility in B. napus. The ratio of SI plants to SC plants from S-1300 × 97-wen135 F₂ and (S-1300 × 97-wen135) × 97-wen135 was 346:260 and 249:232, fitting the expected ratio of 9:7 and 1:1, respectively. Based on these observations, here we propose a genetic model in which two independent loci, S locus and S suppressor locus (sp), are predicted to control the inheritance of maintenance for self-incompatibility in B. napus. The genotypes of S-1300 and 97-wen135 are S ₁₃₀₀ S ₁₃₀₀ sp ₁₃₀₀ sp ₁₃₀₀ and S ₁₃₅ S ₁₃₅ sp ₁₃₅ sp ₁₃₅ , respectively. S ₁₃₅ is dominant to S ₁₃₀₀ , but coexistence of sp ₁₃₀₀ and sp ₁₃₅ fails to suppress S locus. Both S ₁₃₀₀ and S ₁₃₅ can be suppressed by sp ₁₃₅ , while sp ₁₃₀₀ can suppress S ₁₃₅ but not S ₁₃₀₀ . The model contains two characteristics: that a dominant S locus exists in self-compatible B. napus, and that co-suppression will occur when sp loci are heterozygous. The model has been validated by the segregation of S phenotypes in the (S-1300 × 97-wen135) × S-1300, the progenies of SC S-1300 × 97-wen135 F₂ plants and DH population developed from S-1300 × 97-wen135 F₁. This is the first study to report co-suppression of S suppressor loci in B. napus. The genetic model will be very useful for developing molecular markers linked to maintenance for self-incompatibility and for dissecting the mechanism of SI/SC in B. napus.     

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.     

SSR markers for marker assisted selection of root-knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) resistant plants in cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L)

Cotton (Gossypium hirsutum L) cultivars highly resistant to the southern root-knot nematode (RKN) [Meloidogyne incognita (Kofoid and White) Chitwood] are not available. Resistant germplasm lines are available; however, the difficulty of selecting true breeding lines has hindered applied breeding and no highly resistant cultivars are available to growers. Recently, molecular markers on chromosomes 11 and 14 have been associated with RKN resistance, thus opening the way for marker assisted selection (MAS) in applied breeding. Our study aimed to determine the utility of these markers for MAS. Cross one was RKN resistant germplasm M240 RNR × the susceptible cultivar, FM966 and is representative of the initial cross a breeder would make to develop a RKN resistant cultivar. Cross two consists of Clevewilt 6 × Mexico Wild (PI563649), which are the two lines originally used to develop the first highly RKN resistant germplasm. Mexico Wild is photoperiodic. We phenotyped the F₂ of cross one for gall index and number of RKN eggs per plant and genotyped each plant for CIR 316 (chromosome 11) and BNL 3661 (chromosome 14). From this, we verified that MAS was effective, and the QTL on chromosome 14 was primarily associated with a dominant RKN resistance gene affecting reproduction. In the first F₂ population of cross two, we used MAS to identify 11 plants homozygous for the markers on chromosomes 11 and 14, and which also flowered in long days. Progeny of these 11 plants were phenotyped for RKN gall index and egg number and confirmed as RKN highly resistant plants. Generally about 7–10 generations of RKN phenotyping and progeny testing were required to develop the original RKN highly resistant germplasms. Our results show that commercial breeders should be able to use the markers in MAS to rapidly develop RKN resistant cultivars.     

Exploitation of the late flowering species <Emphasis Type="Italic">Brassica oleracea</Emphasis> L. for the improvement of earliness in <Emphasis Type="Italic">B. napus</Emphasis> L.: an untraditional approach

The oilseed Brassica rapa flowers and matures earlier than B. oleracea, as well as their amphidiploid B. napus. Therefore, earliness of B. rapa has been investigated as a source of variation for earliness in B. napus breeding programs. Variation for days to flower exists in B. oleracea; however, its earliest flowering variant B. alboglabra flowers 2–3 weeks later than B. napus. We hypothesized that the C genome of B. alboglabra carries alleles for early flowering which are different from the C-genome alleles of B. napus; and these alleles can be used for the improvement of B. napus. To test this, we examined flowering time in pedigree and DH populations from two B. napus × B. alboglabra crosses. A B. napus line with about a week earlier flowering than the B. napus parent was achieved through reconstitution of its C genome following pedigree selection. Introgression of the B. alboglabra allele in the early flowering pedigree lines is also evident from the presence of B. alboglabra-specific SSR alleles in this line. However, application of doubled haploidy failed to generate any line that flowered earlier than the B. napus parent, which is probably due to the difficulty of obtaining large numbers of euploid B. napus DH lines from this interspecific cross. Thus, we demonstrate that a trait of the diploid species, which apparently looks undesirable, might in fact be highly valuable for the improvement of amphidiploids; and knowledge from this research can also be applied for other traits.     

Genetics of resistance to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis> var. <Emphasis Type="Italic">hardwickii</Emphasis> R. Alef

The genetics of resistance to Cucumber mosaic virus (CMV) in Cucumis sativus var. hardwickii R. Alef, the wild progenitor of cultivated cucumber was assessed by challenge inoculation and by natural infection of CMV. Among the 31 genotypes of C. sativus var. hardwickii collected from 21 locations in India the lowest mean percent disease intensity (PDI) was recorded in IC-277048 (6.33%) while the highest PDI was observed in IC-331631 (75.33%). All the four cultivated varieties (DC-1, DC-2, CHC-1 and CHC-2) showed very high PDI and susceptible disease reaction. Based on mean PDI, 8 genotypes were categorized as resistant, 13 as moderately resistant, 9 as moderately susceptible and one as susceptible. A chi-square test of frequency distribution based on mean PDI in F₂ progenies of six resistant × susceptible crosses revealed monogenic recessive Mendelian ratio 1(R):3(S) to be the best fit. This monogenic recessive model was further confirmed by 1(R):1(S) ratio as the best fit for back cross with resistant parent and no fit for either 3:1 or 1:1 in the back cross with the susceptible parent. The results revealed that CMV resistance in C. sativus var. hardwickii was controlled by a single recessive gene. Considering the cross compatibility between C. sativus var. hardwickii and cultivated cucumber, the resistance trait can be easily transferred to cultivated species through simple backcross breeding.     

Screening global <Emphasis Type="Italic">Medicago</Emphasis> germplasm for weevil (<Emphasis Type="Italic">Hypera postica</Emphasis> Gyll.) tolerance and estimation of genetic variability using molecular markers

The genus Medicago encompasses many important forage species for both temperate and tropical regions. M. sativa L., commonly known as lucerne, is one of the most important forage species grown worldwide, but its production suffers seriously from weevil (Hypera postica Gyll.) infestation. The aim of this work was to identify species/accessions with tolerance to weevil and their molecular analysis using simple sequence repeat (SSR) markers. After screening 197 global germplasm encompassing 50 Medicago species for weevil tolerance, 22 lines representing 13 species were identified where leaf damage was ≤15% (P ≤ 0.05). In total, 37 accessions of the 22 lines, five Indian lucerne cultivars with leaf damage ≥75% and 10 accessions of the 13 Medicago species with low to high infestation (>25%) were molecularly assessed using 11 SSR markers (5 newly developed) to delineate closest to lucerne lines for breeding. In total, 33 bands were scored. The SAHN clustering using UPGM algorithm resulted into two main clusters supported with high boot strap values and with genetic similarity ranging from 0.33 to 0.96. Two accessions of M. tenoreana were observed closest to Indian lucerne cultivars. The rich variability revealed can be used as potential resource for transferring genes across species. Although the inter-specific hybridization is difficult preposition in genus Medicago largely due to post fertilization barrier, the identified species/accessions can be utilized on priority in breeding programs especially employing biotechnological tools like culturing of fertilized pods, ovule-embryo culture and electroporation.     

Production of mutants with high cold tolerance in spring canola (<Emphasis Type="Italic">Brassica napus</Emphasis>)

A major factor affecting spring canola (Brassica napus) production in Canada is killing frosts during seedling development in the spring and seed maturation in the fall. The objective of this study was to explore the possibility of producing spring canola lines with mutations that have altered biochemical pathways that increase cold tolerance. The approach was to generate UV point mutations in cultured microspores followed by chemical in vitro selection of individual mutant microspores or embryos resulting in measurable alterations to various biochemical pathways with elevated levels of key defense signaling molecules such as, salicylic acid (SA), p-Fluoro-d,l-Phenyl Alanine (FPA), and jasmonic acid (JA). In addition, since proline (Pro) is known to protect plant tissues in the cold-induced osmotic stress pathway, mutants that overproduce Pro were selected in vitro by using three Pro analogues: hydroxyproline (HP), azetidine-2-carboxylate (A2C); and, 3,4-dehydro-d,l-proline (DP). Of the 329 in vitro selected mutant embryos produced, 74 were identified with significant cold tolerance compared to their donor parents through indoor freezer tests at −6°C, and 19 had better winter field survival than winter canola checks. All chemically selected mutant doubled haploids with increased cold tolerance compared well with parent lines for all seed quality and agronomic parameters. Development of increased frost tolerant cultivars should allow for spring canola to be produced in western Canada without compromising seed quality.     

Identification and genetics of resistance to cercospora leaf spot (<Emphasis Type="Italic">Cercospora zonata</Emphasis>) in faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis>)

The fungal disease cercospora leaf spot CLS (Cercospora zonata) has affected major faba bean (Vicia faba) production regions in southern Australian in the last several years. This study offers the first report of sources of resistance to CLS in faba bean and describes techniques to evaluate resistance to C. zonata in faba bean genotypes within a controlled environment. The method was rapid (43 days), repeatable (R ² > 0.74) and demonstrated positive correlations (R ² > 0.45–0.80) to data collected from field disease nurseries under naturally established CLS epiphytotics. All faba bean cultivars currently adopted by the Australian industry were found to be susceptible to CLS and defoliation was found to be an important component of disease expression. Genetic analysis of segregation patterns in F ₂ derived F ₃ families of 1322/2*Farah (resistant*susceptible) showed the mode of inheritance of resistance to C. zonata was monogenic dominant. F ₃ families were shown to segregate in the ratio of 1:2:1 for homozygous resistant: heterozygous: homozygous susceptible (χ₂² = 2.78; P > 0.05) and individual plants within heterozygous F ₃ families segregated in the ratio of 3:1 for resistant: susceptible responses (χ₁² = 2.93; P > 0.05). Monogenic dominant inheritance also explained the change in frequency of resistant and susceptible plants within a population of cv. Cairo following one generation of self-pollination (χ² = 0.88, 0.3 < P < 0.5). The sources of resistance identified in this study are being used to transfer CLS resistance to adapted faba bean genotypes for future cultivar releases to the southern Australian industry.     

Identification of resistant sources against <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> in <Emphasis Type="Italic">Brassica</Emphasis> species with emphasis on <Emphasis Type="Italic">B. oleracea</Emphasis>

Stem rot caused by Sclerotinia sclerotiorum is one of the most devastating diseases of rapeseed (Brassica napus L.) which causes huge loss in rapeseed production. Genetic sources with high level of resistance has not been found in rapeseed. In this study, 68 accessions in six Brassica species, including 47 accessions of B. oleracea, were evaluated for leaf and stem resistance to S. sclerotiorum. Large variation of resistance was found in Brassica, with maximum differences of 5- and 57-folds in leaf and stem resistance respectively. B. oleracea, especially its wild types such as B. rupestris, B. incana, B. insularis, and B. villosa showed high level of resistance. Our data suggest that wild types of B. oleracea possess tremendous potential for improving S. sclerotiorum resistance of rapeseed.     

Genetic and histological characterization of a novel recessive genic male sterile line of <Emphasis Type="Italic">Brassica napus</Emphasis> derived from a cross with <Emphasis Type="Italic">Capsella bursa</Emphasis>-<Emphasis Type="Italic">pastoris</Emphasis>

In a previously made cross Brassica napus cv. Oro (2n = 38) × Capsella bursa-pastoris (2n = 4x = 32), one F₁ hybrid with 2n = 38 was totally male sterile. The hybrid contained no complete chromosomes from C. bursa-pastoris, but some specific AFLP (amplified fragment length polymorphism) bands of C. bursa-pastoris were detected. The hybrid was morphologically quite similar to ‘Oro’ except for smaller flowers with rudimentary stamens but normal pistils, and showed good seed-set after pollination by ‘Oro’ and other B. napus cultivars. The fertility segregation ratios (3:1, 1:1) in its progenies indicated that the male sterility was controlled by a single recessive gene. In the pollen mother cells of the male sterile hybrid, chromosome pairing and segregation were normal. Histological sectioning of its anthers showed that the tapetum was multiple layers and was hypertrophic from the stage of sporogenic cells, and that the tetrads were compressed by the vacuolated and disaggregated tapetum and no mature pollen grains were formed in anther sacs, thus resulting in male sterility. The possible mechanisms for the production of the male sterile hybrid and its potential in breeding are discussed.     

Pre and posthaustorial resistance to rusts in <Emphasis Type="Italic">Lathyrus cicera</Emphasis> L.

Lathyrus cicera has a high potential as fodder crop in dry areas, but can in particular environments be damaged by rust. Little is known on the availability of resistance against rust fungi and the underlying mechanisms in L. cicera germplasm. The present study assessed and characterised macro and microscopically the resistance to rust fungi Uromyces pisi and U. viciae-fabae, in a collection of L. cicera accessions. A wide range of disease reaction was found in the germplasm collection against the different rust species. L. cicera accessions were highly resistant to U. viciae-fabae being hypersensitive response the most frequent reaction. On the contrary, most accessions showed a compatible interaction with U. pisi, with varying levels of partial resistance, although cases of hypersensitivity were also identified. Differences on germination, orientated germ tube growth and appressoria differentiation were observed but were in general of marginal importance to explain the resistance to U. pisi among the L. cicera accessions. Resistance was due, to a combination of pre and post-haustorial mechanisms.     

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.     

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.     

Genetic analysis of resistance to <Emphasis Type="Italic">soil-borne wheat mosaic virus</Emphasis> derived from <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Genetic Analysis of Resistance to Soil-Borne Wheat Mosaic Virus Derived from Aegilops tauschii. Euphytica. Soil-Borne Wheat Mosaic Virus (SBWMV), vectored by the soil inhabiting organism Polymyxa graminis, causes damage to wheat (Triticum aestivum) yields in most of the wheat growing regions of the world. In localized fields, the entire crop may be lost to the virus. Although many winter wheat cultivars contain resistance to SBWMV, the inheritance of resistance is poorly understood. A linkage analysis of a segregating recombinant inbred line population from the cross KS96WGRC40 × Wichita identified a gene of major effect conferring resistance to SBWMV in the germplasm KS96WGRC40. The SBWMV resistance gene within KS96WGRC40 was derived from accession TA2397 of Aegilops taushcii and is located on the long arm of chromosome 5D, flanked by microsatellite markers Xcfd10 and Xbarc144. The relationship of this locus with a previously identified QTL for SBWMV resistance and the Sbm1 gene conferring resistance to soil-borne cereal mosaic virus is not known, but suggests that a gene on 5DL conferring resistance to both viruses may be present in T. aestivum, as well as the D-genome donor Ae. tauschii.     

Increased resistance to <Emphasis Type="Italic">Ustilago zeae</Emphasis> and <Emphasis Type="Italic">Fusarium verticilliodes</Emphasis> in maize inbred lines bred for <Emphasis Type="Italic">Fusarium graminearum</Emphasis> resistance

Preliminary field observations in our maize breeding nurseries indicated that breeding for improved resistance to gibberella ear rot (Fusarium graminearum) in maize may indirectly select for resistance to another ear disease, common smut (Ustilago zeae). To investigate this, we compared the disease severity ratings obtained on 189 maize inbreds, eight of which included our inbreds developed with selection for gibberella ear rot resistance after field inoculation and breeding for 8–10 years. No correlation was found between disease severities for the 189 inbreds but the eight gibberella-resistant lines were consistently more resistant to smut. To further examine this relationship and to determine if these eight inbreds would be useful for developing inbreds with either common smut or fusarium ear rot (F. verticilliodes) resistance, we conducted a Griffing’s diallel analysis on six inbreds of maize, four with high levels of gibberella ear rot resistance representing all of the pedigree groups in our eight gibberella lines, and two with very low levels. Our most gibberella ear rot resistant inbreds, CO433 and CO441, had the lowest disease ratings for all three diseases, the consistently largest general combining ability effects and several significant specific combining ability effects. It was concluded that some inbreds bred specifically for gibberella ear rot would also be useful in breeding for resistance to common smut and fusarium ear rot.     

Identification of levels of resistance to cassava root rot disease (<Emphasis Type="Italic">Botryodiplodia theobromae</Emphasis>) in African landraces and improved germplasm using <Emphasis Type="Italic">in vitro</Emphasis> inoculation method

Cassava root rot disease is an increasing problem in Africa where yield losses of about 80% have been recorded. We evaluated 290 African landraces and 306 improved genotypes from the germplasm collections of the International Institute of Tropical Agriculture (IITA), for sources of resistance using root slice laboratory assay. Disease severity was assessed quantitatively by direct percentage estimation (PS) and by use of a rating scale (RS). Both methods of assessment were compared for identification of variability in the germplasm, and genotypes were classified into response groups using an enlarged rank-sum method that combined the PS and RS assessments. The two scoring methods revealed continuous variation (P < 0.001) for resistance in the sets of germplasm. Disease assessments based on PS and RS were highly correlated in both the improved germplasm (r = 0.75) and the landraces (r = 0.72). Based on PS assessment, 50 improved genotypes (16.3%) and 53 landraces (18.3%) showed significantly lower disease scores than the resistant control. The rank-sum method separated each set of collections into highly resistant, resistant, moderately resistant, moderately susceptible, susceptible and highly susceptible groups. Fifty-nine improved genotypes (16.4%) and 61 African landraces (16.9%) were identified as either highly resistant or resistant. Generally, these genotypes exhibited resistance by limiting the growth of the pathogen (reduced amount of invaded surface area). This type of rate-reducing resistance is highly heritable and a quantitative trait which can be harnessed in breeding. Genotypes subsets were identified for further studies into the genetic basis of resistance to root rot disease.     

Germplasm evaluation and transfer of Verticillium wilt resistance from Pima (<Emphasis Type="Italic">Gossypium barbadense</Emphasis>) to Upland cotton (<Emphasis Type="Italic">G. hirsutum</Emphasis>)

Verticillium wilt (VW, Verticillium dahliae) is a worldwide destructive soil-borne fungal disease and employment of VW resistant cultivars is the most economic and efficient method in sustainable cotton production. However, information concerning VW resistance in current commercial cotton cultivars and transfer of VW resistance from Pima (Gossypium barbadense) to Upland (Gossypium hirsutum) cotton is lacking. The objective of the current study was to report findings in evaluating commercial cotton cultivars and germplasm lines for VW resistance in field and greenhouse (GH) experiments conducted in 2003, 2006, and 2007. In the study, 267 cultivars and germplasm lines were screened in the GH, while 357 genotypes were screened in the field. The results indicated that (1) VW significantly reduced cotton yield, lint percentage, 50% span length and micronaire, but not 2.5% span length and fiber strength, when healthy and diseased plants in 23 cultivars were compared; (2) some commercial cotton cultivars developed by major cotton seed companies in the US displayed good VW resistance; (3) many Acala cotton cultivars released in the past also had good VW resistance, but not all Acala cotton germplasm are resistant; (4) Pima cotton possessed higher levels of VW resistance than Upland cotton, but the performance was reversed when the root system was wounded after inoculation; (5) VW resistance in some conventional cultivars was transferred into their transgenic version through backcrossing; and (6) some advanced backcross inbred lines developed from a cross between Upland and Pima cotton showed good VW resistance. The successful development of VW resistant transgenic cultivars and transfer of VW resistance from Pima to Upland cotton implies that VW resistance is associated with a few genes if not a major one.     

Microsporogenesis of <Emphasis Type="Italic">Rps</Emphasis>8/<Emphasis Type="Italic">rps</Emphasis>8 heterozygous soybean lines

Phytophthora root and stem rot caused by Phytophthora sojae, is one of the most damaging diseases of soybean, for which management is principally done by planting resistant cultivars with race specific resistance which are conferred by Rps (Resistance to Phytophthora sojae) genes. The Rps8 locus, identified in the South Korean landrace PI 399073, is located in a 2.23 Mbp region on soybean chromosome 13. In eight cv. Williams (rps8/rps8) × PI 399073 (Rps8/Rps8) populations, this region exhibited strong segregation distortion. In a cross between the South Korean lines PI 399073 (Rps8/Rps8) and PI 408211B (multiple Rps genes) this region segregated in a Mendelian fashion. In this study, microsporogenesis was evaluated to identify meiotic abnormalities that may be associated with the segregation distortion of the Rps8 region. Pollen was collected from greenhouse-grown plants of the parental genotypes: Williams, PI 399073, and PI 408211B; as well as selected Rps8/rps8 RILs from Williams × PI 399073 BC₄F_2:3 and PI 399073 × PI 408211B F_4:5 populations. There were no differences for pollen viability among the genotypes. However, for PI 399073, a mix of dyads, triads, tetrads and pentads was observed. A high frequency of meiotic abnormalities including fragments, laggards, multinucleated microspores; and microcytes containing DNA was also observed in Rps8/rps8 Williams × PI 399073 BC₄F_2:3 RILs. These meiotic abnormalities may contribute to the high degree of segregation distortion present in the Williams × PI 399073 populations.