Quantitative trait locus mapping for Verticillium wilt resistance in a backcross inbred line population of cotton (Gossypium hirsutum × Gossypium barbadense) based on RGA-AFLP analysis

Verticillium wilt (VW), caused by Verticillium dahliae Kleb., is one of the most important diseases in cotton. The objective of this study was to map quantitative trait loci (QTLs) conferring VW resistance using resistance gene analog (RGA)-targeted amplified fragment length polymorphism (RGA-AFLP) markers in an interspecific backcross inbred line mapping population, consisting of 146 lines from a susceptible Sure-Grow 747 (Gossypium hirsutum L.) × resistant Pima S-7 (G. barbadense L.) cross. VW resistance was evaluated in replicated tests based on disease incidence in the field, and disease incidence and severity in the greenhouse. Of 160 polymorphic RGA-AFLP markers, 42 were significantly correlated with one or more VW traits and 41 were placed on a linkage map which covered 1,226 cM of the cotton genome and contained 251 other molecular markers. Three QTLs for VW resistance were detected, each of which explained 12.0–18.6 % of the phenotypic variation. Two of these QTLs for disease incidence and severity detected in the greenhouse inoculation tests using root wounding are located on chromosome c4. Both are closely linked to four RGA-AFLP markers and therefore considered as the same QTL for VW resistance. The other QTL detected in the field test was located on c19 and flanked by several RGA-AFLP markers. The desirable QTL allele on c4 for VW resistance detected in the greenhouse was from the VW susceptible Upland parent and absent from the resistant Pima parent which was more VW susceptible due to the disarmament of the first line of defense mechanism due to root wounding during inoculation. The other desirable VW resistance QTL allele, on c19, was from the resistant parent Pima S-7, consistent with the fact that Pima cotton was more resistant to VW when naturally infected in the field. The results should facilitate the development of more sequence specific markers and the transfer of VW resistance from Pima to Upland cotton through marker-assisted selection.     

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.     

Introgression of <Emphasis Type="Italic">Gossypium barbadense</Emphasis> L. into Upland cotton germplasm RMBUP-C4S1

Gossypium barbadense L. cotton has significantly better fiber quality than Upland cotton (G. hirsutum L.); however, yield and environmental adaptation of G. barbadense is not as wide as Upland. Most cotton in the world is planted to Upland cultivars. Many attempts have been made, over a considerable number of years, to introgress fiber quality alleles from G. barbadense into Upland. However, introgression barriers, primarily in the form of interspecific incompatibility, have limited these traditional approaches. The use of chromosome substitution lines (CSL) as a bridge should provide a more efficient way to introgress alleles from G. barbadense into Upland. We crossed 18 G. barbadense CSL to three cultivars and developed a random mated population. After five cycles of random mating followed by one generation of self-pollination to increase the seed supply, we grew the random mated population and used 139 G. barbadense chromosome specific SSR markers to assess a random sample of 96 plants for introgression. We recovered 121 of 139 marker loci among the 96 plants. The distribution of the G. barbadense alleles ranged from 10 to 28 alleles in each plant. Among the 96 plants we found individual plants with marker loci from 6 to 14 chromosomes or chromosome arms. Identity by descent showed little relatedness among plants and no population structure was indicated by a heat map. Using CSL we were able to develop a mostly Upland random mated population with considerable introgression of G. barbadense alleles which should be useful for breeding.     

Evaluation of Verticillium wilt resistance in commercial cultivars and advanced breeding lines of cotton

Verticillium wilt (VW), caused by Verticillium dahliae Kleb, is one of the most destructive diseases in cotton (Gossypium spp.). The most efficient and cost-effective method of controlling the disease is the use of resistant cotton cultivars. Most commercial cultivars and elite breeding lines are developed under non-VW conditions and their responses to the disease are currently unknown. This study was conducted to evaluate current commercial cotton cultivars and advanced breeding lines for VW resistance. In 2011–2013, a total of 84 cultivars from major US seed companies, 52 advanced breeding lines from the US public breeding programs, and 87 introgression lines from a cross between Acala 1517-99 × Pima PHY 76 from the New Mexico Cotton Breeding Program, were evaluated for VW resistance in the greenhouse. Cotton cultivars and breeding lines were evaluated in ten separate replicated tests by inoculation with a defoliating-type isolate of V. dahliae. While leaf severity rating and percentages of infected plants, infected leaves and defoliated leaves were found to be significantly and positively correlated with one another, leaf severity rating and percentage of infected leaves were best choices because of their relatively low coefficients of variation and higher resolutions to differentiate resistant genotypes from susceptible ones. The heritabilities for the VW resistance traits ranged from 0.58 to 0.80 with an average of 0.67, indicating that variation in VW resistance is predominantly due to genetic factors. Of the 223 commercial cultivars and advanced lines, six Upland cultivars (FM 9160B2F, FM 9170 B2F, NG 4010 B2RF, Nitro 44 B2RF, DP 1219 B2RF, and ST 4288 B2F), five advanced lines (Ark 0403-3, MD 10-5, MD 25ne, NC11AZ01, and PD 0504), two introgression lines from Upland × Pima (NM11Q1157 and 08N1618), and four Pima cultivars (COBALT, DP 357, PHY 800, and PHY 830) had higher levels of resistance to VW. The resistance shown by most of these cultivars in the greenhouse was consistent with their performance in previous field tests. Based on the initial VW resistance, 19 highly or moderately resistant genotypes were chosen for re-evaluation and 30 genotypes were also assessed more than once for VW resistance in different tests, most of which had concordant performance. These cultivars and advanced lines should be useful resources to improve VW resistance in cotton breeding.     

Morph-physiological responses of cotton interspecific chromosome substitution lines to low temperature and drought stresses

Limited knowledge about genetic and physiological traits associated with drought and low temperature stresses and narrow genetic diversity in Upland cotton (Gossypium hirsutum L.) are serious impediments in its genetic improvement. The objectives of this research were to determine the genetic and physiological traits associated with drought and low temperature effects and to identify chromosomal effects on these traits using chromosome substitution (CS) lines from three alien species of Gossypium, G. barbadense, G. tomentosum, and G. mustelinum, respectively. Two experiments were conducted to study low temperature and drought stress effects during seedling emergence and early growth stages in 21 cotton CS-lines with parent, Texas Marker (TM)-1. In Experiment I, plants were grown at optimum (30/22 °C) and low (22/14 °C) temperature conditions under optimum water and nutrient conditions. In Experiment II, plants were grown at optimum water (soil moisture content of 0.167 m³ m^?3) and in drought (soil moisture content 0.105 m³ m^?3) conditions under optimum temperature conditions. Above- and below-ground growth traits including several root traits of the CS lines were assessed at 25 days after sowing. The findings suggest which substituted chromosome or chromosome segment from the alien species likely harbors one or more genes for higher and lower tolerance to low temperature, respectively. CS-T04 and CSB08sh showed higher and lower tolerance to low temperature, respectively and CS-T04 and CS-B22sh showed higher and lower tolerance, respectively, to drought. CS lines are valuable analytical tool and useful genetic resources for targeted exploitation of beneficial genes for drought and low temperature stresses in Upland cotton.     

High-density genetic map construction and QTL mapping for fiber strength on Chr24 across multiple environments in a CCRI70 recombinant inbred lines population

Upland cotton is an important economic crop that produces high-quality fiber for the textile industry. With the development of next-generation sequencing technology and improvements in human living standards, it has become possible to improve the fiber quality and yield of cotton with high-throughput molecular markers. Upland cotton 901-001 is an excellent, high-quality, non-transgenic cultivar, while the sGK156 strain shows high resistance to verticillium wilt. The phenotype of F₁ plants, certified in 2008 as national variety CCRI70, shows positive transgressive characteristics such as high quality, high yield, and resistance to verticillium wilt. We developed a population of 250 recombination inbred lines from a cross between 901-001 and sGK156. The fiber strength trait of plants from nine environments was collected, and a genetic linkage map of Chr24 comprising 168 SNP marker loci covering a genetic distance of 107.46 cM and with an average distance of 0.64 cM was generated. QTLs were identified across the nine environments using the composite interval mapping method. A total of eight QTLs for FS were identified on Chr24, three of which were stably expressed in at least five environments. Some candidate genes located in qFS-c24-2 and qFS-c24-4 were functionally annotated as potentially playing important roles in fiber development, with homologous genes reported in Arabidopsis thaliana. These results suggest that QTLs identified in the present study could contribute to improving FS and may be applicable for marker-assisted selection.     

Quantitative trait locus mapping of drought and salt tolerance in an introgressed recombinant inbred line population of Upland cotton under the greenhouse and field conditions

Drought and salt tolerances are complex traits and controlled by multiple genes, environmental factors and their interactions. Drought and salt stresses can result in more than 50% yield loss in Upland cotton (Gossypium hirsutum L.). G. barbadense L. (the source of Pima cotton) carries desirable traits such as tolerance to abiotic and biotic stress along with high fiber quality. However, few studies have been reported on mapping quantitative trait loci (QTL) for abiotic stress tolerance using a permanent bi-parental population in multiple tests. The transfer of drought and salt tolerance from Pima to Upland cotton has been a challenge due to interspecific hybrid breakdown. This issue may be overcome by using introgression lines with genes transferred from Pima to Upland cotton. In this study, four replicated tests were conducted in the greenhouse each for drought and salt tolerance along with another test conducted in a field for drought tolerance using an Upland recombinant inbred line population of TM-1/NM24016 that has a stable introgression from Pima cotton. The objectives of the study were to investigate the genetic basis of drought and salt tolerance and to identify genetic markers associated with the abiotic stress tolerance. A total of 1004 polymorphic DNA marker loci including RGA-AFLP, SSR and GBS-SNP markers were used to construct a genetic map spanning 2221.28 cM. This population together with its two parents was evaluated for morphological, physiological, yield and fiber quality traits. The results showed that drought under greenhouse and field conditions and salt stress in the greenhouse reduced cotton plant growth at the seedling stage, and decreased lint yield and fiber quality traits in the field. A total of 165 QTL for salt and drought tolerance were detected on most of the cotton chromosomes, each explaining 5.98–21.43% of the phenotypic variation. Among these, common QTL for salt and drought tolerance were detected under both the greenhouse and field conditions. This study represents the first study to report consistent abiotic stress tolerance QTL from multiple tests in the greenhouse and the field that will be useful to understand the genetic basis of drought and salt tolerance and to breeding for abiotic stress tolerance using molecular marker-assisted selection in cotton.     

Genotypic comparisons of chromosomes 01, 04, and 18 from three tetraploid species of <Emphasis Type="Italic">Gossypium</Emphasis> in topcrosses with five elite cultivars of <Emphasis Type="Italic">G. hirsutum</Emphasis> L.

Upland cotton, Gossypium hirsutum L. is the most widely planted cultivated cotton in the United States and the world. The other cultivated tetraploid species G. barbadense L. is planted on considerable less area; however, it produces extra long, strong, and fine fibers which spins into superior yarn. The wild cotton tetraploid species G. tomentosum Nuttall ex Seemann, native to the Hawaiian Archipelago also exhibits traits, such as drought tolerance, that would also be desirable to transfer to Upland cotton. Long-term breeding efforts using whole genome crosses between Upland and these species have not been successful in transferring very many desirable alleles into Upland cotton. Our chromosome substitution lines (CSL) have one chromosome or chromosome arm from an alien species backcrossed into the Upland cotton line,TM-1, via aneuploid technology. Five Upland cultivars were crossed with CS-B01, CS-T01, CS-B04, CS-T04, CS-B18 and CS-T18 and TM-1 the recurrent parent of the CSLs. This provided an opportunity to determine the effects of chromosomes 01, 04, and 18 from the three species in crosses with the five cultivars. Predicted genotypic mean effects of the parents, F₂, and F₃ generations for eight agronomic and fiber traits of importance were compared. The predicted hybrid mean effects for the three chromosomes from each species were different for several of the traits across cultivars. There was no single chromosome or species that was superior for all traits in crosses. Parental and hybrid lines often differed in the effect of a particular chromosome among the three species. The predicted genotypic mean effects for F₂ and F₃, with a few exceptions, generally agree with our previous results for additive and dominance genetic effects of these CSL.     

Targeted transfer of trait for Verticillium wilt resistance from Gossypium barbadense into G. hirsutum using SSR markers

Verticillium wilt (VW) is a soil‐borne disease of cotton that is destructive worldwide. Transferring desired traits from Gossypium barbadense is challenging through traditional interspecific introgression. We previously demonstrated that a molecular marker, BNL3255‐208, is associated with VW resistance in G. barbadense. This breakthrough opens the way for marker‐assisted selection (MAS) breeding. Here, the highly resistant G. barbadense cv. ‘Pima90‐53’ and the severe diseased Gossypium hirsutum cv. ‘CCRI8’ were used as donor parent and recipient parent, respectively. Our goal was to transfer the disease resistance from donor to recipient via MAS. Among 71 MAS obtained lines, as many as 19 lines had enhanced resistance. Among those lines, 11 lines showed high resistance and four lines displayed resistance to VW. Moreover, seven lines displayed improved fibre quality. After combining the markedly improved resistance and fibre properties, we identified two elite innovated introgression lines – ZY2 and ZY31 – that did not seem to differ in other agronomic traits from the recipient parent. This study first successfully transferred of G. barbadense resistance into G. hirsutum by MAS.     

Hybridization between cotton and Malvaceae species as a tool for production of partial interspecific aneuploid cotton plants

Cotton although is an autogamous species could be cross pollinated under favorable climate conditions and/or in the presence of pollinators. The coexistence of cotton with Malvaceae species raises questions on the possibility pollen to be exchanged among Malvaceae species and on the resulted consequences. The present work was undertaken to evaluate the in situ response of cotton flowers (G.hirsutum L, G. barbadense L.) and their F₁ interspecific hybrids when are artificially pollinated with Malva sylvestris L., Hibiscus syriacus L. and Abelmoschus esculentus Moench. Furthermore, an in vitro protocol was attempted to support embryos’ growth in order to produce viable progenies originating from crosses between cotton and the aforementioned Malvaceae species. The obtained results gave evidence that pollen from the above Malvaceae species stimulated cotton ovaries without successful hybridization. The interaction between pollen and cotton’s stigmas was higher at early stages when M. sylvestris was used as pollinator but in crosses with H. syriacus and A. esculentus more ovules were activated as revealed by the percentage of carpodesis and life-time of bolls onto maternal plants. Only crosses between cotton with okra produced cotton seeds under in situ conditions. The in vitro embryo-ovule culture protocol, used, increased the number of regenerated cotton plants, especially in crosses among F₁ interspecific cotton hybrids and A. esculentus. In this case, regenerated plants were recombinant aneuploids, combining traits from both cotton species. This novel cotton germplasm possessing unique chromosome rearrangements, at aneuploid level could be proved useful after cytogenetic, molecular or QTL genetic analysis referring to important agronomic traits.     

Determining resistance to <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> and fumonisin accumulation in African maize inbred lines resistant to <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and aflatoxins

Fusarium verticillioides and Aspergillus flavus cause Fusarium ear rot (FER) and Aspergillus ear rot (AER) of maize, respectively. Both pathogens are of concern to producers as they reduce grain yield and affect quality. F. verticillioides and A. flavus also contaminate maize grain with the mycotoxins fumonisins and aflatoxins, respectively, which has been associated with mycotoxicosis in humans and animals. The occurrence of common resistance mechanisms to FER and AER has been reported. Hence, ten Kenyan inbred lines resistant to AER and aflatoxin accumulation were evaluated for resistance to FER, F. verticillioides colonisation and fumonisin accumulation; and compared to nine South African lines resistant to FER and fumonisin accumulation. Field trials were conducted at three localities in South Africa and two localities in Kenya. FER severity was determined by visual assessment, while F. verticillioides colonisation and fumonisin content were quantified by real-time PCR and liquid chromatography tandem mass spectrometry, respectively. Significant genotype x environment interactions was determined at each location (P ≤ 0.05). Kenyan inbred CML495 was most resistant to FER and F. verticillioides colonisation, and accumulated the lowest concentration of fumonisins across localities. It was, however, not significantly more resistant than Kenyan lines CML264 and CKL05015, and the South African line RO549 W, which also exhibited low FER severity (≤5%), fungal target DNA (≤0.025 ng μL^?1) and fumonisin levels (≤2.5 mg kg^?1). Inbred lines resistant to AER and aflatoxin accumulation appear to be promising sources of resistance to F. verticillioides and fumonisin contamination.     

Dynamic QTL analysis and validation for plant height using maternal and paternal backcrossing populations in Upland cotton

Plant height determines plant biomass yield, harvest index and economic yield. We analyzed quantitative trait loci (QTL) and gene action controlling plant height. We generated the maternal and paternal testcrossing (TC/M and TC/P) populations based on a recombinant inbred line population. Data for plant height at t1, t2, t3, t4 or t5 stages were collected over 2 years from 3 TC/M field trials and 2 TC/P field trials. At single-locus level, 32 QTLs at five stages and 24 conditional QTLs at four intervals were detected, and 14 QTLs shared in different years or populations or stages. Plant height displayed dynamic characteristics through expression of QTLs. A total of 21 novel QTLs were detected and 11 QTLs validated the previous results. And 19 QTLs explained over 10% of phenotypic variation, such as qPH-Chr9-2, qPH-Chr19-4 and qPH-Chr22-4. The region of NAU5330-NAU1269 on chromosome 19 may be a desired target for genetic improvement of plant height in Upland cotton. In addition, five and eight heterotic loci were identified in TC/M and TC/P populations, respectively. Additive, partial dominance and overdominance effects were observed in both TC populations. We also identified 43 epistatic QTLs and QTLs by environment interactions by inclusive composite interval mapping method. Taken together, additive, partial dominance and overdominance effects together with epistasis explained the genetic basis of plant height in Upland cotton.     

Simple genetic inheritance conditions resistance to <Emphasis Type="Italic">Liriomyza sativae</Emphasis> in melon

The leafminer Liriomyza sativae (Diptera: Agromyzidae) stands out as the main plant health problem in melon in the Northeast region of Brazil, which is the main region for production and export of the fruit. Genetic resistance of plants is an important strategy in management of this pest. The plant BAGMEL 56-R was selected as a new source of resistance to L. sativae through antibiosis; this resistance is characterized by the death of larvae soon after they begin feeding on the leaf mesophyll; the result is leaf mines that are small and insignificant in terms of yield reduction. Lines with contrasting levels of resistance were obtained from the progenies of this source of resistance through successive self-pollinations, conducted by the pedigree breeding method. Through the segregation pattern of the progenies and the test cross, the genetic nature of resistance was determined; one gene with complete dominance conditions resistance. The name Liriomyza sativae resistance and the symbol Ls are suggested to represent this new gene. In addition, through a non-preference test with lines in contrast for antibiosis and the susceptible hybrid ‘Goldex’, the presence of antixenosis was observed in this source of resistance. Probably, these different types of resistance in the source BAGMEL 56-R are associated with distinct defense mechanisms. Therefore, with this new source, introgression of resistance to L. sativae in elite lines or commercial hybrids of melon is possible.     

Stability of <Emphasis Type="Italic">Solanum aethiopicum</Emphasis> Shum accessions under varied water deficit stress levels and identification of pertinent breeding traits for resistance to water shortage

Drought is a major constraint to productivity of Solanum aethiopicum ‘Shum’ group due to loss in market and nutrient value of stressed plants. This study evaluated S. aethiopicum Shum group accessions to identify genotypes (G) that excel across moisture deficit stress levels (WLs). A split-plot arrangement composed of four WLs and twenty accessions of S. aethiopicum as main plot and sub-plot factors, respectively, was implemented in a screenhouse, and repeated for two experiments. In each experiment, there was a highly significant effect of at least two WLs on mean performance among at least two accessions for most of the traits at p < 0.05. Further, very highly significant WL × G interactions were obtained for leaf relative water content (LRWC), leaves per plant (LPP) and plant height (PH), and non-significant for leaf blade length and leaf blade width. The order of priority as breeding traits for stability superiority across WLs was suggested as LRWC > PH > LPP. Consequently, based on LRWC, the most superiorly stable accessions were identified as accession 160 followed by accessions 145, 137, 108P and 184G while the least stable ones were identified as accessions 163G, 141, 163 and 108. The broad sense heritability (H ²) for each of the three recommended traits for drought resistance breeding was above 0.9 thus supportive for a good response to selection. Drought stress negatively affected the performance of S. aethiopicum Shum group but the exhibited variation allowed for selection of superiorly stable genotypes.     

Inheritance of foliar resistance to <Emphasis Type="Italic">Phytophthora</Emphasis> species in cacao (<Emphasis Type="Italic">Theobroma cacao</Emphasis> L.): can it be influenced by CSSV infection?

Studies on quantitative genetics of foliar resistance to black pod disease in cacao could inadvertently use cocoa swollen shoot virus (CSSV) infected leaves which could bias the results especially in West Africa where the virus is prevalent. However, effects of CSSV on inheritance and heritability of foliar resistance to Phytophthora species is not known. Choice of an efficient breeding method requires an accurate estimation of genetic effects in selection schemes for foliar resistance to Phytophthora species in cacao. The objective of this study was to investigate the effect of CSSV infection on quantitative genetic parameters of foliar resistance to cocoa black pod disease in a population of 36 F1 hybrids developed by mating six cacao genotypes using a diallel method. The generated F1s and their parents were evaluated for foliar resistance to P. palmivora and P. megakarya using a randomized complete block design (RCBD) with three replications. 1A CSSV and Nsaba CSSV strains were used to infect the cacao genotypes using the patch graft method. The parents chosen showed significant variations for scores of leaf discs after inoculation with P. palmivora and P. megakarya. The leaf disc scores of CSSV infected crosses were lower than leaf disc scores of CSSV-free crosses. Genetic component analysis showed that the effects of GCA and SCA was significant for both CSSV-free and CSSV-infected crosses in resistance to P. palmivora and P. megakarya. The significant GCA and SCA for both CSSV-free and CSSV-infected crosses strongly suggest that both additive and non-additive genetic effects play an important role in the determination of inheritance of foliar resistance to Phytophthora species in cacao. There was significant variability in mean squares of GCA and SCA of CSSV-free and CSSV-infected crosses indicating that CSSV infection modifies GCA and SCA of affected plants. Narrow sense heritability was relatively low (0.31) for foliar resistance to P. palmivora and P. megakarya under CSSV-free and 1A CSSV strain infected conditions. However, heritability for foliar resistance to P. palmivora (0.43) and P. megakarya (0.36) was significantly higher under Nsaba CSSV infected condition. The modifications of mean squares of GCA and SCA and narrow sense heritability due to CSSV infection could mislead in choice of breeding methods indicating that attention must be paid to the infection status of plants when conducting quantitative genetics studies using diseased and healthy plants. CSSV status of leaf samples should be known before using them in leaf disc test. Genotypes Pa7/808 and Pound 7 had high negative GCA effects and are promising parents for enhancement of resistance to black pod disease in cacao.     

Resistance to <Emphasis Type="Italic">Cucumber green mottle mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Cucumber green mottle mosaic virus (CGMMV) is a severe threat for cucumber production worldwide. At present, there are no cultivars available in the market which show an effective resistance or tolerance to CGMMV infection, only wild Cucumis species were reported as resistant. Germplasm accessions of Cucumis sativus, as well as C. anguria and C. metuliferus, were mechanically infected with the European and Asian strains of CGMMV and screened for resistance, by scoring symptom severity, and conventional RT-PCR. The viral loads of both CGMMV strains were determined in a selected number of genotypes using quantitative RT-PCR. Severe symptoms were found following inoculation in C. metuliferus and in 44 C. sativus accessions, including C. sativus var. hardwickii. Ten C. sativus accessions, including C. sativus var. sikkimensis, showed intermediate symptoms and only 2 C. sativus accessions showed mild symptoms. C. anguria was resistant to both strains of CGMMV because no symptoms were expressed and the virus was not detected in systemic leaves. High amounts of virus were found in plants showing severe symptoms, whereas low viral amounts found in those with mild symptoms. In addition, the viral amounts detected in plants which showed intermediate symptoms at 23 and 33 dpi, were significantly higher in plants inoculated with the Asian CGMMV strain than those with the European strain. This difference was statistically significant. Also, the amounts of virus detected over time in plants did not change significantly. Finally, the two newly identified partially resistant C. sativus accessions may well be candidates for breeding programs and reduce the losses produced by CGMMV with resistant commercial cultivars.     

Heritability of the resistance to pepper huasteco yellow vein virus in wild genotypes of <Emphasis Type="Italic">Capsicum annuum</Emphasis>

Pepper huasteco yellow vein virus (PHYVV) is the main virus of pepper crop in Mexico. No resistant cultivars are available and resistance breeding is hampered by the lack of knowledge of heritability (h ²) of PHYVV resistance. This is a continuation of previous studies and the objectives were to analyze the h ² and the behavior of the resistant trait to PHYVV. Four resistant assays were done with three resistant wild lines (UAS12, UAS13 and UAS10) of Capsicum annuum in the S4, S5, S6 and S7 generation under greenhouse conditions. Plants from all tests were inoculated with PHYVV through Bemisia tabaci. Line UAS12 was the most resistant showing a significantly proportion of resistant plants, less disease symptoms and longer incubation time, followed by the lines UAS13 and UAS10 in all assays. Distribution of symptoms showed a bimodal tendency in all the trials, suggesting that two groups of genes are involved in this resistance trait. The lines UAS12, UAS13 and UAS10 showed the same pattern of response to selection with an average of h ² of 0.17, 0.06, 0.02 and 0.00 in the S4, S5, S6 and S7, respectively. These results indicate that all lines responded positively to the selection in the S4, S5 and S6, whereas in the S7 there was no response by the possible exhaustion of variation. Line UAS12 is the most promising genotype and the lines UAS13 and UAS10 are genetic resources that can be supplemented to breed the resistance of PHYVV. These results provides basic information for resistance breeding.     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

A SNP mutation affects rhizomania-virus content of sugar beets grown on resistance-breaking soils

Rhizomania is one of the most devastating biotic stresses affecting sugar beet (Beta vulgaris L.). It is caused by Beet necrotic yellow vein virus (BNYVV) vectored by the plasmodiophorid Polymyxa betae K. The only means available to control the disease is the use of genetically resistant varieties. “Rizor” or “Holly” (Rz1) and WB42 (Rz2) have been the most widely used resistance sources in the commercial varieties. Recently, naturally occurring resistance-breaking (RB) rhizomania strains have been identified causing major concerns. The aim of this study was to identify SNP mutations that show associations with resistance to rhizomania in sugar beet plants grown under resistance-breaking (RB)-BNYVV soils. Rhizomania virus content was evaluated by indirect triple-antibody sandwich-ELISA within two F ₂ segregating populations respectively grown on an AYPR and IV-BNYVV strain infected soils. Bulked segregant analysis (BSA) was performed. The resistant and susceptible plants were genotyped with a 384-SNPs panel. Of the 384 SNPs, SNP249 was found to associate with the resistance both to the AYPR strain (R ² = 0.37; P = 0.0004) and to the IV-BNYVV (R ² = 0.09; P = 0.0074). Our results suggested that the SNP249 could be readily applicable for marker-assisted breeding of resistance to AYPR strain of rhizomania.     

Genetic variability of faba bean genotypes for chocolate spot (<Emphasis Type="Italic">Botrytis fabae</Emphasis>) resistance and yield

Faba bean (Vicia faba L.) has high utility as a food and soil fertility improving crop. One of the major fungal pathogens of faba bean is Botrytis fabae, the causative agent of chocolate spot. The disease affects significantly the leaf, stem, pod and seed of faba bean compromise its productivity in the smallholder farming sector. Nonetheless, there are limited resistant/tolerant faba bean varieties available and disease control technology options. Therefore, it was prudent to evaluate faba bean landraces for chocolate spot resistance. Fifty landraces together with ten improved varieties were evaluated both in the field and in the greenhouse under natural and artificial inoculation with previously selected aggressive Botrytis fabae isolate (Iso-016) from West Gojjam, in Ethiopia. There were highly significant differences (p?<?0.001) among the landraces for reaction to the disease and agronomic traits. Significant positive correlation was recorded between reaction of genotypes in the field and greenhouse disease data. The overall mean disease epidemics varied from 92.5 to 697.5 for the area under disease progress curve (AUDPC). The highest level of resistance was found in the ICARDA lines, ILB-4726, ILB-938 and BPL-710. Of all 18 landrace collections displayed significantly lower disease reaction than the susceptible check. However the resistance was moderate. The selected eighteen landraces will be recommended for use in breeding for chocolate resistance. Overall, resistance was highly heritable, suggesting that phenotypic selection can be exploited to improve chocolate spot resistance in faba bean varieties.