Phenotypic variability of Leptosphaeria lindquistii (anamorph: Phoma macdonaldii), a fungal pathogen of sunflower

Growth of 17 isolates of Phoma macdonaldii , the causal agent of sunflower black stem, was investigated for response to pH and temperature, and for morphology and asexual morphogenesis (pycnidiogenesis and pycnidium size). For all isolates, the optimum pH for growth was between 4 and 5, and the optimum temperature varied between 20 and 30°C and radial growth was slowest at 5 and 35°C. Significant differences in the number and size of pycnidia were observed between isolates. Pycniospore germination was investigated under various conditions in five isolates chosen for their geographical origins, pigmentation, optimum growth temperature and pycnidiogenesis. Increasing the concentration from 10⁶ to 10⁷ pycniospores per mL decreased the germination rate. The optimum temperature for pycniospore germination varied between 15 and 30°C, depending on the isolate, and the optimum and maximum pH values were 5 and 7, respectively. The optimum and minimum relative humidities allowing pycniospore germination were 100 and 95%, respectively. Pycniospore germination was photo-independent. An artificial inoculation method was developed and the aggressiveness of the pathogen was assessed on a susceptible sunflower cultivar, using a 1–9 scale that integrated the percentage of necrotic area on the cotyledon petiole at the stage when the first pair of leaves was fully developed. Significant differences in aggressiveness were observed among the 17 isolates. The parameters investigated clearly suggest the occurrence of a wide phenotypic variability in Phoma macdonaldii .     

Genotype-isolate interaction for resistance to black stem in sunflower (Helianthus annuus)

Two experiments were undertaken to determine the partial resistance of sunflower genotypes to seven isolates of Phoma macdonaldii . In the first experiment, 28 genotypes, including recombinant inbred lines and their parents, M6 mutant lines developed by gamma irradiation, and some genotypes from different geographical origins, were used. The experiment consisted of a split-plot design with three replications, each with 12 seedlings per genotype per isolate, in controlled conditions. Seven days after inoculation, plantlets were scored on a 1–9 scale for percentage necrotic area. Highly significant differences were observed among genotypes, isolates and their interactions. The presence of a differential interaction between genotypes and P. macdonaldii isolates was confirmed in a second experiment using 12 genotypes representing large variability for partial resistance to P. macdonaldii isolates, as identified in the first experiment. Inbred lines B454/03, ENSAT-B5 and LC1064C were the most susceptible sunflower genotypes, whereas two American lines SDR19 and SDR18 presented high partial resistance to all P. macdonaldii isolates studied. The least and most aggressive isolates were MA6 and MP3, respectively. Isolates interacted differentially with sunflower genotypes. This study identified two genotypes (AS613 and PAC2) presenting specific resistance to isolate MP8. The results also showed a wide range of isolate-nonspecific partial resistances among the lines tested. The information presented here could assist sunflower breeders to choose parents of crosses for breeding of durable resistance to phoma black stem disease.     

法国向日葵种子中向日葵黑茎病菌的首次截获与检测

从法国进境向日葵种子中分离到3株疑似向日葵黑茎病的菌株,对所有菌株进行形态学、致病性测定和分子序列比对分析。分离菌菌落乳白色或象牙色至灰白色,有大量黑褐色分生孢子器产生,分生孢子器球形至扁球形,内含无色单胞、卵圆形分生孢子,有明显或不明显油球;针刺接种4片真叶向日葵幼苗的下胚轴,7～9d后茎部产生典型黑茎病黑色椭圆形病斑,病斑上着生黑色分生孢子器;菌丝DNA用ActF1/R1和ITS1/ITS4扩增,序列与NCBI基因库中P.macdonaldii序列相似性为98%～100%。形态学、分子生物学及致病性检测结果显示,截获的3株菌均为向日葵黑茎病菌。     

Quantitative trait loci associated with resistance to a potato isolate of Verticillium albo‐atrum in Medicago truncatula

Verticillium albo‐atrum is responsible for considerable yield losses in many economically important crops, among them alfalfa (Medicago sativa). Using Medicago truncatula as a model for studying resistance and susceptibility to V. albo‐atrum, previous work has identified genetic variability and major resistance quantitative trait loci (QTLs) to Verticillium. In order to study the genetic control of resistance to a non‐legume isolate of this pathogen, a population of recombinant inbred lines (RILs) from a cross between resistant line F83005.5 and susceptible line A17 was inoculated with a potato isolate of V. albo‐atrum, LPP0323. High genetic variability and transgressive segregation for resistance to LPP0323 were observed among RILs. Heritabilites were found to be 0·63 for area under the disease progress curve (AUDPC) and 0·93 for maximum symptom score (MSS). A set of four QTLs associated with resistance towards LPP0323 was detected for the parameters MSS and AUDPC. The phenotypic variance explained by each QTL (R²) was moderate, ranging from 4 to 21%. Additive gene effects showed that favourable alleles for resistance all came from the resistant parent. The four QTLs are distinct from those described for an alfalfa V. albo‐atrum isolate, confirming the existence of several resistance mechanisms in this species. None of the QTLs co‐localized with regions involved in resistance against other pathogens in M. truncatula.     

Inheritance of resistance to stem canker (Phomopsis helianthi) in sunflower

Six male sterile sunflower lines were crossed with seven restorers in a factorial mating design. The 13 inbred lines and their 42 F1 hybrids were planted in a randomized block design with three replicates. Each replicate consisted of two rows, 5 m long (30–35 plants per replicate). Resistance to natural Phomopsis infection, presented as the percentage of plants with no encircling necrosis lesions of the fungus on the main stem, was determined at physiological maturity. Analysis of variance showed that female and male general combining abilities (GCA) and specific combining abilities (SCA) of F1 hybrids were significant. The ratio of additive variance to total variance was 0.662, a high value which indicates prevailing additive effects. The additive variance due to females was more important than that of males, probably because of the existence of maternal effects or more effective genes for resistance in the female lines used in this experiment. The estimates of GCA were significant and positive for LC1004A, KO549A, 50KD8 and LC1064C inbred lines. These lines should be considered in developing hybrids with improved resistance to Phomopsis in sunflower breeding programmes.     

Locating quantitative trait loci associated with Orobanche crenata resistance in pea

Orobanche crenata (broomrape) is a root parasite that represents the major constraint for pea (Pisum sativum) cultivation in the Mediterranean area and the Middle East. The efficacy of available control methods is minimal and breeding for O. crenata resistance is considered the most promising strategy of control. Only moderate levels of incomplete resistance are available in pea germplasm. In order to identify and map the quantitative trait loci (QTL) controlling the trait, 115 F₂ plants derived from the cross between a susceptible and a resistant parent were analysed using isozymes, random amplified polymorphic DNA and sequence tagged site markers. F₂‐derived F₃ lines were studied for O. crenata resistance under field conditions. The linkage map was constructed with MAPMAKER V2.0. Of 217 markers, 120 could be mapped into 21 linkage groups. Linkage groups consisted of 13 to two marker loci covering 1770 cM. The mean inter‐marker distance was 17.64 cM. Simple interval mapping (SIM) and composite interval mapping (CIM) were performed using QTL Cartographer. The CIM approach using five cofactors was clearly the most efficient way to locate putative QTL. Two QTL for O. crenata resistance (Ocp1 and Ocp2), explaining only a moderate portion of the observed variation (9.6% and 11.4% respectively), were detected.     

Methodology of virulence screening and race characterization of Plasmopara halstedii,and resistance evaluation in sunflower – a review

Sunflower downy mildew is a disease of high global economic impact as well as a causal agent that is extremely difficult to eradicate. During the past decades, several approaches for the determination of Plasmopara halstedii (Ph) races have been used worldwide and are discussed in this review. Procedures of isolation, cultivation and maintenance of Ph isolates, as well as the screening of sunflower for resistance, are also critically reviewed. The predominant, globally used resistance screening protocol is a ‘whole seedling immersion’ inoculation. ‘Soil drench’ inoculation allows more precise control of the number of Ph zoosporangia applied to a single sunflower seedling. A detached leaf assay has been described, but it has been used mainly for Ph subcultivation and fungicide tests. For race determination, a differential set consisting of nine sunflower genotypes has been used since 1988, coupled with a numerical triplet code for virulence phenotyping of Ph. The increasing variability in global Ph populations has demonstrated the inadequacy of the current set of differentials, and several researchers have proposed additional public lines as new differentials. Furthermore, bulk isolates may show different results in repeated tests, as Ph may contain genetically distinct zoospores within a single zoosporangium. For precise race determination, single zoosporangia or single zoospore isolates are advisable. However, due to low success of isolation, approximately 1–2%, this method cannot be applied in routine Ph race screening. Methods surveyed in this review have a broad spectrum of applications, including taxonomic studies.     

Selection for quantitative trait loci associated with resistance to Stewart's wilt in sweet corn

The objectives of this research were to identify quantitative trait loci (QTL) for Stewart's wilt resistance from a mapping population derived from a sweet corn hybrid that is highly resistant to Pantoea stewartii and to determine if marker-based selection for those QTL could substantially improve Stewart's wilt resistance in a population derived from a cross of resistant lines and a highly susceptible sweet corn inbred. Three significant QTL for Stewart's wilt resistance on chromosomes 2 (bin 2.03), 5 (bin 5.03), and 6 (bin 6.06/6.07) explained 31% of the genetic variance in a population of 110 F(3:4) families derived from the sweet corn hybrid Bonus. The three QTL appeared to be additive in their effects on Stewart's wilt ratings. Based on means of families that were either homozygous or heterozygous for marker alleles associated with the resistance QTL, the QTL on chromosomes 2 and 6 appeared to have dominant or partially dominant gene action, while the QTL on chromosome 5 appeared to be recessive. A population of 422 BC(2)S(2) families was derived from crosses of a sweet corn inbred highly susceptible to Stewart's wilt, Green Giant Code 88 (GG88), and plants from two F(3:4) families (12465 and 12467) from the Bonus mapping population that were homozygous for marker alleles associated with Stewart's wilt resistance at the three QTL. Mean Stewart's wilt ratings for BC(2)S(2) families were significantly (P < 0.05) lower for families that were homozygous for the bnlg1902 marker allele (bin 5.03) from resistant lines 12465 or 12467 than for families that were heterozygous at this marker locus or homozygous for the bnlg1902 marker allele from GG88. Resistance associated with this QTL was expressed only if F(3:5) or BC(2)S(2) families were homozygous for marker alleles associated with the resistant inbred parent (P(1)). Marker alleles identified in the F(3:5) mapping population that were in proximity to the resistance QTL on chromosomes 2 and 6 were not polymorphic in crosses of GG88 with 12465 and 12467. Selection for other polymorphic marker loci adjacent to these two regions did not improve Stewart's wilt resistance of BC(2)S(2) families.     

Quantitative trait loci for seedling and adult plant resistance to Stagonospora nodorum in wheat

Stagonospora nodorum blotch (SNB) caused by Stagonospora nodorum is a severe disease of wheat (Triticum aestivum) in many areas of the world. S. nodorum affects both seedling and adult plants causing necrosis of leaf and glume tissue, inhibiting photosynthetic capabilities, and reducing grain yield. The aims of this study were to evaluate disease response of 280 doubled haploid (DH) individuals derived from a cross between resistant (6HRWSN125) and susceptible (WAWHT2074) genotypes, compare quantitative trait loci (QTL) for seedling and adult plant resistance in two consecutive years, and assess the contribution of QTL on grain weight. Flag leaves and glumes of individuals from the DH population were inoculated with mixed isolates of S. nodorum at similar maturity time to provide accurate disease evaluation independent of morphological traits and identify true resistance for QTL analysis. Fungicide protected and inoculated plots were used to measure relative grain weight (RGW) as a yield-related trait under pathogen infection. The lack of similar QTL and little or no correlation in disease scores indicate different genes control seedling and adult plant disease and independent genes control flag leaf and glume resistance. This study consistently identified a QTL on chromosome 2DL for flag leaf resistance (QSnl.daw-2D) and 4BL for glume resistance (QSng.daw-4B) from the resistant parent, 6HRWSN125, explaining 4 to 19% of the phenotypic variation at each locus. A total of 5 QTL for RGW were consistently detected, where two were in the same marker interval for QSnl.daw-2D and QSng.daw-4B indicating the contribution of these QTL to yield related traits. Therefore, RGW measurement in QTL analysis could be used as a reliable indicator of grain yield affected by S. nodorum infection.     

Genetic control of partial resistance to ‘collar’ and ‘root’ isolates of <Emphasis Type="Italic">Phoma macdonaldii</Emphasis> in sunflower

Phoma macdonaldii is one of the most important pathogens of sunflower (Heliantus annuus) in France. In order to determine the inheritance of resistance to the disease, five sunflower genotypes with wide genetic variability for resistance to two ‘collar’ and two ‘root’ Phoma isolates were crossed in a diallel programme. Four separate experiments were undertaken under controlled conditions. In each one, the response of parental genotypes and their F1 hybrids were evaluated with one of the four Phoma isolates. Analysis of variance was performed to determine the effects of genotype on disease severity score when inoculated with ‘collar’ or ‘root’ Phoma isolates and showed significant variability among parents and F1 hybrids for disease severity score. Diallel analysis showed that general combining ability (GCA) and specific combining ability (SCA) effects for resistance to ‘collar’ and ‘root’ Phoma isolates were highly significant for each of the four isolates indicating that both kinds of gene effects were important in controlling the resistance. The GCA/SCA ratios were more than one for three out of four isolates showing that additive genetic effects were more important than non-additive effects for resistance to three of the studied Phoma isolates. Hence, conventional breeding methods could be recommended to achieve genetic improvement to such ‘collar’ and ‘root’ Phoma isolates.     

Quantitative trait loci for resistance to fusarium head blight and deoxynivalenol accumulation in Wangshuibai wheat under field conditions

Wangshuibai is a Chinese landrace wheat with a high level of resistance to fusarium head blight (FHB) and deoxynivalenol (DON) accumulation. Using an F7 population of recombinant inbred lines (RILs) derived from the cross between Wangshuibai and Annong 8455 for molecular mapping of quantitative trait loci (QTL) for FHB resistance, the proportion of scabbed spikelets (PSS) and DON content were assessed under field conditions. Composite interval mapping revealed that two and three QTL were significantly associated with low PSS and low DON content, respectively, over 2 years. QTL on chromosomes 3B and 2A explained 17 and 11·5%, respectively, of the phenotypic variance for low PSS, whereas QTL on chromosomes 5A, 2A and 3B explained 12·4, 8·5 and 6·2%, respectively, of the phenotypic variance for low DON content. The 3B QTL appeared to be associated mainly with low PSS, and the 5A QTL primarily with low DON content in Wangshuibai. The 2A QTL had minor effects on both low PSS and DON content. Microsatellite and AFLP markers linked to these QTL should be useful for marker-assisted selection of QTL for low PSS and low DNA content from Wangshuibai.     

Identification of isolate-specific and partial resistance to septoria tritici blotch in 238 European wheat cultivars and breeding lines

From a total of 238 European cultivars and breeding lines screened for isolate-specific resistance to septoria tritici blotch (STB) with eight Mycosphaerella graminicola isolates from five different countries, 142 lines were resistant to Ethiopian isolate IPO88004, and 43 lines were specifically resistant to IPO323, with little or no leaf area bearing pycnidia of M. graminicola . These lines probably all have the resistance gene Stb6 . Specific resistances to isolates CA30JI, IPO001, IPO89011, IPO92006 and ISR398 were less common. Seventy-three per cent of the lines were specifically resistant to at least one isolate and 36 lines were resistant to more than one isolate. The line with the greatest number of specific resistances was the spring cultivar Raffles, with five. The most resistant line in which no specific resistance was identified was the Italian landrace Rieti, an ancestor of many modern European wheat cultivars. There was also a wide range of partial resistance among the lines tested, expressed in detached seedling leaves. Information about the resistance of wheat lines to M. graminicola isolates will assist breeders to choose parents of crosses from which progeny with superior resistance to STB may be selected.     

Notes on the biology and ecology of Ptecticus testaceus (Diptera: Stratiomyidae) a soldier fly associated with mango fruit (m. indica) in Costa Rica

Abstract

The association between Ptecticus testaceus Fabr. (Dipt.: Stratiomyidae) and mango fruits attached to the tree is reported, with some observations on the ecology and behaviour on the soldier fly. P.testaceus apparently synchronized its biological cycle to mango fruit set in Costa Rica, and its adults feed on nectar of sunflower inflorescences. More detailed studies to determine the status of this dipteran, and its association with mango fruits, are suggested.     

Strategy to identify resistance to white mould associated with high yield for irrigated common bean in Brazil

Development of common bean cultivars with partial white mould resistance through breeding techniques has been a challenge in Brazil. As yet, lines/cultivars from breeding programmes have not been investigated for resistance; therefore, this study screened 107 lines/cultivars for their reactions to white mould in 14 preliminary trials conducted under irrigation. Thirteen resistant lines/cultivars (three of Andean origin) and six Mesoamerican cultivars (three intermediately resistant and three susceptible) were selected for further investigation. These lines/cultivars and the resistant control A195 were evaluated in six advanced trials and two straw tests to assess the effectiveness of the screening procedure. In 11 preliminary trials, screenings were performed under moderate/high or higher disease pressure. These pressures occurred in two advanced trials in which, when yields were averaged across moderate/high and high pressures, 10 Mesoamerican lines/cultivars selected for resistance yielded 14%, 23%, and 38% more than intermediately resistant cultivars, A195, and susceptible cultivars, with median disease ratings (1–9 scale) of 4.5, 5.7, 5.7, and 6.7, respectively. In the straw test, three Andean lines/cultivars (A195 included) and two susceptible cultivars in the field were among those with the highest levels of physiological resistance. Thus, field rating under high disease pressure and greenhouse rating did not correlate significantly, suggesting that field trials are critical to evaluating resistance and to identifying high-yielding beans. Therefore, lines/cultivars from breeding programmes assessed in field trials may provide a low cost and fast way to identify high-yielding bean cultivars with partial resistance to white mould in the subtropical southern hemisphere.     

Susceptibility of sunflower inbreds to Melanagromyza minimoides in Argentina and potential association with plant resistance traits

Seed-feeding by larvae of the mosquito head fly, Melanagromyza minimoides, can substantially reduce yields of late-planted sunflower in South America and its management with insecticides or early-planting has other undesirable costs. Evaluation of inbred lines over three seasons indicated differences in the emergence of adult flies, particularly between restorer (male) and maintainer (female) lines used for hybrid seed production; when fly emergence (≈damage) was adjusted for differences in head size (i.e. to flies/100 cm²), restorer lines averaged 3–12 times more damage than maintainer lines. Fly emergence in each of two seasons was tested for correlations with three sunflower traits (glandular trichome number, pericarp strength, and sesquiterpene lactone content) believed to provide host plant resistance to other insects. In one of two years, fly emergence was negatively correlated with the relative amount of sesquiterpene lactones in florets of the tested lines. Also, among maintainer lines, severity of infestation was much greater in multi-headed inbreds compared to single-headed lines. Additional research is needed to establish the cause(s) of reduced susceptibility of inbreds and whether parental lines influence M. minimoides damage to hybrids.     

Mating and dispersal behaviour of Scirpophaga incertulas and Chilo suppressalis (Lepidoptera; Pyralidae) in relation to resistance management for rice transformed with Bacillus thuringiensis toxin genes

Information on the mating and dispersal behaviour of the rice stem borers Scirpophaga incertulas and Chilo suppressalis is necessary for the design of resistance management strategies for rice varieties genetically engineered with toxin genes from Bacillus thuringiensis (Bt). We investigated whether S. incertulas and C. suppressalis move from the site of adult eclosion before mating to determine the likelihood that random mating will occur between adults eclosing in fields of Bt rice and refuge fields of non-Bt rice. Observations were made under laboratory, screenhouse and field conditions. Almost all S. incertulas males and females flew out of sight after cuticle expansion and hardening, as did most C. suppressalis. Approximately 15% of C. suppressalis females called and mated within 3?m of the site of eclosion, and approximately 5% of C. suppressalis males mated within 5?m of the site of eclosion. These results suggest that random mating is likely to occur between S. incertulas eclosing in Bt rice fields and nearby refuge fields. Although a small proportion of C. suppressalis did not move far before mating, random mating of moths from Bt and nearby refuge fields is also likely to occur in this species if Bt-resistant moths are rare.