Identification and marker‐assisted introgression of QTL conferring resistance to bacterial leaf blight in cowpea (Vigna unguiculata (L.) Walp.)

Bacterial leaf blight (BLB), caused by Xanthomonas axonopodis pv. vignicola (Xav), is widespread in major cowpea [Vigna unguiculata (L.) Walp.] growing regions of the world. Considering the resource poor nature of cowpea farmers, development and introduction of cultivars resistant to the disease is the best option. Identification of DNA markers and marker‐assisted selection will increase precision of breeding for resistance to diseases like bacterial leaf blight. Hence, an attempt was made to detect QTL for resistance to BLB using 194 F_2 : 3 progeny derived from the cross ‘C‐152’ (susceptible parent) × ‘V‐16’ (resistant parent). These progeny were screened for resistance to bacterial blight by the leaf inoculation method. Platykurtic distribution of per cent disease index scores indicated quantitative inheritance of resistance to bacterial leaf blight. A genetic map with 96 markers (79 SSR and 17 CISP) constructed from the 194 F₂ individuals was used to perform QTL analysis. Out of three major QTL identified, one was on LG 8 (qtlblb‐1) and two on LG 11 (qtlblb‐2 and qtlblb‐3). The PCR product generated by the primer VuMt337 encoded for RIN2‐like mRNA that positively regulate RPM1‐ and RPS2‐dependent hypersensitive response. The QTL qtlblb‐1 explained 30.58% phenotypic variation followed by qtlblb‐2 and qtlblb‐3 with 10.77% and 10.63%, respectively. The major QTL region on LG 8 was introgressed from cultivar V‐16 into the bacterial leaf blight susceptible variety C‐152 through marker‐assisted backcrossing (MABC).     

Identification and characterization of resistance to rust (Uromyces ciceris-arietini (Grognot) Jacz. & Boyd) in a germplasm collection of Cicer spp.

A collection consisting of 140 accessions of chickpea (Cicer arietinum L.) and 109 accessions of related wild species (Cicer spp.) was screened for resistance to chickpea rust (Uromyces ciceris-arietini (Grognot) Jacz. & Boyd). Varying levels of partial resistance were identified, based on a reduced disease severity and area under disease progress curve without any macroscopically visible host cell necrosis. Higher levels of resistance were observed in wild Cicer species, but neither associated with hypersensitivity. Components of resistance have been macros- and microscopically studied in selected C. arietinum accessions. Resistance was expressed as longer latent period and lower infection frequency, which were associated with increased percentage of early aborted colonies, reduced number of haustorial mother cells and haustoria per colony, and reduced colony size.     

Leaf Cell‐Wall Components as Influenced in the First Phase of Salt Stress in Three Maize (Zea mays L.) Hybrids Differing in Salt Resistance

The leaf cell wall (CW) chemical composition of three maize (Zea mays L.) hybrids (salt‐resistant SR 03 and SR 12, salt‐sensitive Pioneer 3906) was investigated in the first phase of salt stress (100 mm NaCl) compared with the control (1 mm NaCl) treatment to investigate whether changes in CW composition were responsible for shoot growth reduction. Salt treatment caused a strong inhibition in shoot growth with a concomitant increase in the ratio between CW dry mass (DM) and shoot fresh mass (FM) and a decrease in CW cellulose concentrations in all hybrids. NaCl caused a large increase in the concentrations of total and non‐methylated uronic acid (UA) in salt‐sensitive Pioneer 3906 and salt‐resistant SR 12. The onset of the accumulation of non‐methylated UA was delayed in SR 12, which indicates that this may be one reason for the better growth performance of this hybrid under salt stress compared with Pioneer 3906. It is concluded that a low accumulation of non‐methylated UA in leaf CW may, among other mechanisms, contribute to salt resistance in the first phase of salt stress.     

Mapping quantitative trait loci (QTLs) underlying seed vitamin E content in soybean with main,epistatic and QTL × environment effects

Soybean (Glycine max (L.) Merr.) seed contains small amounts of tocopherol, a non‐enzymatic antioxidant known as lipid‐soluble vitamin E (VE). Dietary VE contributes to a decreased risk of chronic diseases in humans and has several beneficial effects on resistance to stress in plants, and increasing VE content is an important breeding goal for increasing the nutritional value of soybean. In this study, quantitative trait loci (QTLs) underlying VE content with main, epistatic and QTL × environment effects were identified in a population of F_5 : 6 recombinant inbred lines from a cross between ‘Hefeng 25’ (a low‐VE cultivar) and ‘OAC Bayfield’ (a high‐VE cultivar). A total of 18 QTLs were detected that showed additive main effects (a) and/or additive × environment interaction effects (ae) in different environments. Moreover, 19 epistatic pairs of QTLs were found to be associated with α‐tocopherol (α‐Toc), γ‐tocopherol (γ‐Toc), δ‐tocopherol (δ‐Toc) and total VE (TE) contents. The QTLs identified in multienvironments could provide more information about QTL by environment interactions and could be useful for the marker‐assistant selection of soybean cultivars with high seed VE contents.     

Identification and characterization of sources of resistance to Erysiphe pisi Syd. in Pisum spp.

A collection of 67 accessions of Pisum species originating from different countries was screened in a glasshouse test for resistance to Erysiphe pisi. All Pisum fulvum accessions were completely resistant. Incomplete resistance was identified in some accessions of P. sativum subsp. sativum var. arvense and P. sativum subsp. elatius and abyssinicum. Microscopy revealed several distinct cellular mechanisms governing resistance. In P. fulvum, it was mainly due to a high frequency of cell death that occurred both as a rapid response to attempted infection and a delayed response that followed colony establishment. Cell death following colony establishment was also key to the incomplete resistance in some accessions of P. sativum subsp. sativum var. arvense. In addition, impaired spore germination, and to a lesser extent appressorium formation, contributed to pre‐penetration resistance in some accessions. In some cases, resistance also retarded colony growth, possibly through effects on haustorial development or function in cells that survived the attack. Thus, these wild pea accessions offer diverse resistances that could be introduced to cultivated peas to increase the efficacy of powdery mildew resistance.     

Virulence characteristics of a new race of the parasitic angiosperm,Striga gesnerioides,from southern Benin on cowpea (Vigna unguiculata)

An in vitro growth system was used to determine the virulence of two samples of Striga gesnerioides from Zakpota in southern Benin. Cowpea variety B301, previously considered resistant to all races of S. gesnerioides, was susceptible to both samples of the parasite. Two other cowpea varieties, 58–57 and IT81D-994, were totally resistant. Resistance in 58–57 was associated with a hypersensitive necrosis of infected roots, whilst IT81D-994 supported production of small S. gesnerioides tubercles with stems which failed to develop. Striga gesnerioides from southern Benin is the fourth race of the parasite to be identified, and the first with virulence on variety B301. The implications for breeding cowpeas with resistance to S. gesnerioides are discussed.Abbreviations IITA International Institute of Tropical Agriculture - LARS Long Ashton Research Station - SAFGRAD Semi-Arid Food Grain Research and Development     

The genetic background modulates the intensity of Rpv3‐dependent downy mildew resistance in grapevine

Grape varieties with resistance to downy mildew (DM) carry alien chromosome segments in Vitis vinifera backgrounds. We previously showed that the largest descent group shares a non‐vinifera haplotype at the locus Rpv3. Here, we performed a common garden experiment with 76 varieties to evaluate the level of field resistance across four years. All varieties exhibited effector‐triggered immunity (ETI)‐associated necrosis. On a scale of 1–9, the median OIV452 value for foliar resistance was 7.1 in the resistant lineage vs. 3.2 in vinifera controls. Genotype, year and their interaction significantly affected the level of resistance. Some resistant genotypes showed high mean values of OIV452 and low variance among years. Other resistant genotypes showed lower mean OIV452 and higher variance. They were capable of activating ETI, but the intensity was inadequate to restrict pathogen growth under highly conducive conditions. Rpv3‐dependent responses were stronger in highly native genetic backgrounds and tended to attenuate in late backcross generations. Genetic backgrounds donated by European winegrapes of the convarietas occidentalis provided on average higher levels of Rpv3 resistance than backgrounds of orientalis table grapes.     

Search for partial resistance against Puccinia hordei in barley landraces from the Fertile Crescent

A collection of 111 barley landraces from the Fertile Crescent was screened for resistance to barley leaf rust in the field and under controlled conditions. Large variation was observed for disease severity under field conditions. Accessions with high resistance because of hypersensitivity were identified. Also segregation was observed in some accessions, with individual plants showing hypersensitive reactions (IT ≤ 6). Partial resistance due to a reduction of infection in spite of a compatible infection was commonly found (19%). Resistance of 12 accessions selected for their low disease severity and high IT, was shown to be due to a prolonged latency period and increased percentage of early aborted colonies not associated with host cell necrosis. A high correlation was observed between the microscopic and macroscopic components of partial resistance.     

Quantitative trait loci for leafhopper (Empoasca fabae and Empoasca kraemeri) resistance and seed weight in the common bean

A population of 108 common bean recombinant inbred lines (RILs) (F_5:6‐9), derived from a leafhopper (Empoasca fabae and E. kraemeri)‐susceptible cultivar (‘Berna’) and a leafhopper‐resistant line (EMP 419) was used to identify molecular markers genetically linked to leafhopper resistance and seed weight. Bulked segregant analysis and quantitative trait analysis identified eight markers that were associated with resistance to E. fabae, and four markers that were associated with E. kraemeri resistance. Three markers were associated with resistance to both species. A partial linkage map of the bean genome was constructed. Composite interval mapping identified quantitative trait loci (QTL) for resistance to both leaf hopper species on core‐map linkage groups B1, B3 and B7. QTL for seed weight were found close to the locus controlling testa colour and an α‐phaseolin gene.     

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

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

Characterization of resistance response of pea (Pisum spp.) against rust (Uromyces pisi)

Components of resistance to the pea rust (Uromyces pisi) were studied at the histological level in seedlings of seven pea (Pisum spp.) accessions showing partial resistance under field conditions. Resistance was characterized macroscopically by an increased latent period and a decreased infection frequency in spite of a compatible interaction (high infection type). Histological investigations revealed little differences among accessions in spore germination and appressoria formation, differences being more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with significant levels of early aborted colonies in accessions IFPI3260, PI347321 and PI347347. Furthermore, a reduction in the number of haustoria per colony, hyphal tips per colony and smaller colony size were found in all genotypes studied. Resistance was not associated with early‐ or late‐acting hypersensitivity in any of the genotypes studied.     

Genetic markers linked to Striga gesnerioides resistance for the improvement of Ghanaian cowpea (Vigna unguiculata) cultivars

The parasitic plant, Striga gesnerioides (Willd.) Vatke, is one of the most important constraints of cowpea production and food security in West Africa. Currently, few Striga resistant cowpea varieties have been developed that are well‐adapted to the dry savannah regions of Ghana. While genes conferring resistance to Striga races SG1, SG3 and SG5 have been mapped, the genetic locus of resistance to the race of Striga found in Ghana (SG‐GH) has not been characterized. Here, we report identification of genetic markers linked to SG‐GH resistance and define the relationship of this locus to SG3 resistance in recombinant inbred line populations generated from crosses between Striga resistant IT97K‐499‐35 and the Striga susceptible varieties Apagbaala and SARC‐LO2. The populations were genotyped with five genetic markers associated with SG3 and SG5 Striga resistance genes and a genetic map was developed. Genes conferring resistance to SG‐GH and SG3 mapped 4.2 cM from each other on chromosome Vu11. The identification of genetic markers linked to SG‐GH resistance will facilitate the marker‐assisted development of high‐quality Striga resistant cowpea varieties in Ghana.     

Introducing host‐plant resistance to anthracnose in Kyrgyz common bean through inoculation‐based and marker‐aided selection

Common bean (Phaseolus vulgaris L.) is one of the most important legume crops for human consumption. However, its grain yield can be reduced by up to 90% by the seedborne disease, anthracnose. Fungicide treatment is costly and time‐consuming. The introduction of host plant resistance against this disease appears, therefore, to be crucial for enhancing the productivity of this crop in Kyrgyzstan. The use of DNA‐based markers in backcrossing programmes may help speed up the breeding for resistance. In this study, we used a combination of inoculation tests and a DNA marker (SCAreoli marker) to track the transfer of host‐plant resistance (Co‐2 gene) from two donor cultivars, ‘Vaillant’ and ‘Flagrano’, to susceptible Kyrgyz cultivars ‘Ryabaya’, ‘Kytayanka’ and ‘Lopatka’, which are widely grown in the country. The segregating offspring were evaluated to test the reliability of the SCAreoli marker as selection aid for host‐plant resistance to anthracnose. Our study showed that a co‐dominant DNA marker can successfully be used in backcross breeding to distinguish segregating material in different market classes of common bean.     

Identification and mapping of adult‐plant stripe rust resistance in soft red winter wheat cultivar ‘USG 3555’

Little is known about the extent or diversity of resistance in soft red winter wheat (Triticum aestivum L.) to stripe rust, caused by the fungal pathogen Puccinia striiformis f.sp. tritici. The soft red winter (SRW) wheat cultivar ‘USG 3555’ has effective adult‐plant resistance to stripe rust, which was characterized in a population derived from ‘USG 3555’/‘Neuse’. The mapping population consisted of 99 recombinant inbred lines, which were evaluated for stripe rust infection type (IT) and severity to race PST‐100 in field trials in North Carolina in 2010 and 2011. Genome‐wide molecular‐marker screenings with 119 simple sequence repeats and 560 Diversity Arrays Technology (DArT) markers were employed to identify quantitative trait loci (QTL) for stripe rust resistance. QTL on chromosomes 1AS, 4BL and 7D of ‘USG 3555’ explained 12.8, 73.0 and 13.6% of the variation in stripe rust IT, and 13.5, 72.3 and 10.5% of the variation in stripe rust severity, respectively. Use of these and additional diagnostic markers for these QTL will facilitate the introgression of this source of stripe rust resistance into SRW wheat lines via marker‐assisted selection.     

Drought responses of above‐ground and below‐ground characteristics in warm‐season turfgrass

Water shortages have become more chronic as periodic droughts prolong and water demand for urban and agricultural use increases. Plant drought responses involve coordinated mechanisms in both above‐ and below‐ground systems, yet most studies lack comparisons of root and canopy responses under water scarcity and recovery. This is particularly true of research focused on warm‐season turfgrasses in sandy soils with extremely low water holding capacity. To address the lack of examination of coordinated stress and recovery responses, this study compared the above‐ and below‐ground plant responses during a dry‐down period of 21 days and recovery among four warm‐season turfgrass species in the field. Canopy drought responses and recovery were quantified using digital image analysis. In situ root images were captured using a minirhizotron camera system. Common bermudagrass [Cynodon dactylon (L.) Pers.] endured the entire drought period without losing 50% green cover while other species lost 50% green cover in 11–34 days predicted from the regression. The interspecific differences in drought resistance were mainly due to root characteristics. Other drought mechanisms appear to be responsible for differences identified in drought resistance between “Zeon” and “Taccoa Green” manilagrass [Zoysia matrella (L.) Merr.]. Recovery was delayed for up to 2 weeks in the second year, warranting further evaluation for turfgrass persistence under long‐term drought. Three‐week drought posed no threat to the survival of zoysiagrass. Species and genotypic variations were found in achieving full post‐recovery, which can be used to develop water conservation strategies and to adjust consumer expectations.     

Evaluation of the Ph‐3 gene‐specific marker developed for marker‐assisted selection of late blight‐resistant tomato

Late blight caused by Phytophthora infestans is one of the most destructive diseases of tomato (Solanum lycopersicum L.) that mainly occurs in cool and wet environments. With the spread of the A2 mating type and new clonal lineages, fewer fungicides provide effective control of the disease, which has increased its worldwide threat. Host resistance could contribute significantly to sustainable disease control. Ph‐3 is a race‐specific late blight resistance gene commonly used in commercial tomato breeding. Availability of precise and easy to use gene‐based markers would facilitate selection. In this study, a Ph‐3 on‐gene cleaved amplified polymorphic sequence (CAPS) marker, Ph3.gsm/HincII, was developed based on the published gene sequence of Ph‐3. The effectiveness of the marker was evaluated along with other published Ph‐3 markers using an F₉ recombinant inbred line (RIL) population derived from NC 23E‐2(93) × L3708. Markers Ph3.gsm/HincII and TG328/BstNI accurately genotyped the RIL population for Ph‐3. In addition, Ph3.gsm/HincII was able to differentiate variable susceptible alleles. This reliable codominant DNA marker would be very useful in marker‐assisted selection, particularly for resistance gene pyramiding.     

Inheritance of resistance to the peach root‐knot nematode (Meloidogyne floridensis) in interspecific crosses between peach (Prunus persica) and its wild relative (Prunus kansuensis)

The peach root‐knot nematode, Meloidogyne floridensis (MF), infects majority of available nematode‐resistant peach rootstocks which are mostly derived from peach (Prunus persica) and Chinese wild peach (P. davidiana). Interspecific hybridization of peach with its wild relative, Kansu peach (P. kansuensis), offers potential for broadening the resistance spectrum in standard peach rootstocks. We investigated the inheritance of resistance to MF in segregating populations of peach (‘Okinawa’ or ‘Flordaguard’) × P. kansuensis. A total of 379 individuals from 13 F₂ and BC₁F₁ families were challenged with a pathogenic MF isolate “MFGnv14” and were classified as resistant (R) or susceptible (S) based on root galling intensity. Segregation analyses in F₂ progeny revealed the involvement of a major locus with a dominant or recessive allele determining resistance in progeny segregating 3R:1S and 1R:3S, respectively. Testcrosses with a homozygous‐susceptible peach genotype (‘Flordaguard’ or ‘UFSharp’) confirmed P. kansuensis as a source of new resistance and the heterozygous allelic status of P. kansuensis at the locus conferring resistance to MF. We propose a single‐locus dominant/recessive model for the inheritance of resistance.     

Adult plant leaf rust resistance from 111 wheat (Triticum aestivum L.) cultivars

Adult plant resistance against Indian leaf rust race 77 and five of its highly virulent variants have been identified from 111 bread wheat cultivars originating from 12 countries. The adult plant resistance of only 16 of these cultivars is due to hypersensitive seedling or adult plant resistance genes. All others expressed nonhypersensitive type of resistance characteristic of the genes Lr34 and Lr46.Forty five of the 111 cultivars showed tip necrosis on flag leaves, a trait linked to the gene Lr34. Therefore, the nonhypersensilive type of resistance of these 45 cultivars is attributed to Lr34. The nonhypersensitive resistance of the remaining cultivars is likely to be due to the gene(s) different than Lr34. The reaction pattern of these 111 cultivars to six races suggests the presence of at least six to seven new hypersensitive adult plant resistance genes and at least three new hypersensitive seedling resistance genes. The known genes Lr10, Lr23 and Lr26 were detected frequently but these genes did not contribute towards the adult plant resistance of any of the 111 cultivars. Based on the presence of new genes for hypersensitive and nonhypersensitive type of resistance, the 111 cultivars have been classified into 31 diverse resistance groups. This revised version was published online in July 2006 with corrections to the Cover Date.     

The resistance to leaf rust and powdery mildew of recombinant lines of barley (Hordeum vulgare L.) derived from H. vulgare × H. bulbosum crosses

A set of 23 recombinant lines (RLs) of barley ( Hordeum vulgare L.) derived from H. vulgare  ×  H. bulbosum L. crosses was inoculated with barley leaf rust ( Puccinia hordei ) and powdery mildew ( Blumeria graminis f.sp. hordei ) at the seedling stage to identify their levels and mechanisms of resistance. Eight RLs were studied further in glasshouse and field tests. All three barley parents ('Emir', 'Golden Promise' and 'Vada') were highly susceptible to powdery mildew and leaf rust isolates. Several RLs showed partial resistance expressed as high relative latency periods and low relative infection frequencies against leaf rust. This high level of partial resistance was due to a very high level of early aborting colonies without host cell necrosis. Several RLs showed hypersensitive resistance to some or all isolates. For powdery mildew, one RL was completely resistant to the CC1 isolate and had a hypersensitive resistance to the CO-02 isolate. Three RLs derived from 'Emir' were completely resistant to both powdery mildew isolates, and three more RLs tested in the field had higher levels of partial resistance than their parents. The results indicate that H. bulbosum contains major and minor gene(s) for resistance to leaf rust and powdery mildew that can be transferred to cultivated barley.     

Pyramiding three rust‐resistance genes confers a high level of resistance in soybean (Glycine max)

Pyramiding Asian soybean rust (ASR) resistance (Rpp) genes in a single genotype has been shown to increase ASR resistance in soybean. However, it remains unclear which combinations of Rpp genes are superior. Therefore, here, we developed six new Rpp‐pyramided lines carrying different combinations of Rpp genes and evaluated their resistance against 13 Bangladeshi rust (Phakopsora pachyrhizi) isolates (BdRPs) alongside seven previously developed Rpp‐pyramided lines. We found that lines carrying one, two and three Rpp genes had high ASR resistance without sporulation in 13.8%, 35.2% and 73.1% of the assays, respectively. Among the new lines that were developed, those with Rpp3 + Rpp4 and Rpp3 + Rpp4 + Rpp5 had high levels of ASR resistance, while the line containing Rpp2 + Rpp4 + Rpp5 showed immunity phenotype at two weeks after inoculation by the BdRP‐22 infection. Thus, pyramiding larger numbers of Rpp genes confers soybean with a higher level of resistance to ASR pathogens and can produce an immunity phenotype at two weeks after inoculation.