The role of selection on the genetic structure of pathogen populations: Evidence from field experiments with Mycosphaerella graminicola on wheat

Summary Coevolution refers to reciprocal genetic changes that occur in two or more ecologically interacting species. In agricultural ecosystems, we are especially concerned with the genetic response of pathogen populations to resistant cultivars produced by plant breeding programs. It would be useful to be able to predict whether disease resistance is likely to be durable or ephemeral before a cultivar is widely grown. Though it may not be possible to predict durability in advance, knowledge of the genetic structure of pathogen populations may prove useful for making predictions about the rate at which pathogens adapt to resistant varieties. Much has been learned about the genetic structure of populations of obligate fungal pathogens such as rusts and mildews, which have become paradigms for plant pathology. We have focused our effort on the population genetics of the less known, non-specialized, necrotrophic pathogens, such as the Septorias of small grains. Our approach has been to use DNA fingerprinting and RFLP analysis to conduct field experiments that elucidate how populations of fungal pathogens adapt in agroecosystems. Our results suggest that mating system may have a greater impact than natural selection on the genetic structure of populations of Mycosphaerella graminicola (anamorph Septoria tritici).     

Screening techniques and sources of resistance against parasitic weeds in grain legumes

Summary A number of parasitic plants have become weeds, posing severe constraints to major crops including grain legumes. Breeding for resistance is acknowledged as the major component of an integrated control strategy. However, resistance against most parasitic weeds is difficult to access, scarce, of complex nature and of low heritability, making breeding for resistance a difficult task. As an exception, resistance against Striga gesnerioides based on a single gene has been identified in cowpea and widely exploited in breeding. In other crops, only moderate to low levels of incomplete resistance of complex inheritance against Orobanche species has been identified. This has made selection more difficult and has slowed down the breeding process, but the quantitative resistance resulting from tedious selection procedures has resulted in the release of cultivars with useful levels of incomplete resistance. Resistance is a multicomponent event, being the result of a battery of escape factors or resistance mechanisms acting at different levels of the infection process. Understanding these will help to detect existing genetic diversity for mechanisms that hamper infection. The combination of different resistance mechanisms into a single cultivar will provide durable resistance in the field. This can be facilitated by the use of in vitro screening methods that allow highly heritable resistance components to be identified, together with adoption of marker-assisted selection techniques.     

Genetic variability for resistance to sorghum downy mildew (Peronosclerospora sorghi) and Rajasthan downy mildew (P. heteropogoni) in the tropical/sub-tropical Asian maize germplasm

Downy mildews cause considerable damage to maize (Zea mays L.) worldwide, particularly in the tropical Asia. We have evaluated a set of 42 tropical/sub-tropical maize inbred lines developed in different countries in Asia (India, Thailand and Philippines), and Mexico, for analysing the genetic variability for resistance to sorghum downy mildew [Peronosclerospora sorghi; SDM] and Rajasthan downy mildew [P. heteropogoni; RDM]. Experiments were carried out in replicated trials under artificial infection in field conditions against SDM and RDM at Mandya in Karnataka, India, and Udaipur in Rajasthan, India, respectively, during 1999 and 2000. The study resulted in identification of five inbred lines offering consistent and strong resistance to both SDM and RDM, while several inbred lines revealed resistance only to RDM. It was also revealed that the SDM-resistant inbreds are invariably resistant to RDM, while the RDM-resistant inbreds might show differential responses to the SDM. The maize inbred lines identified in this study with broad-spectrum resistance to downy mildews could be potentially useful for basic and applied research work on downy mildews in tropical Asia. This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening techniques and sources of resistance to foliar diseases caused by major necrotrophic fungi in grain legumes

Summary Necrotrophic pathogens of the cool season food legumes (pea, lentil, chickpea, faba bean and lupin) cause wide spread disease and severe crop losses throughout the world. Environmental conditions play an important role in the development and spread of these diseases. Form of inoculum, inoculum concentration and physiological plant growth stage all affect the degree of infection and the amount of crop loss. Measures to control these diseases have relied on identification of resistant germplasm and development of resistant varieties through screening in the field and in controlled environments. Procedures for screening and scoring germplasm and breeding lines for resistance have lacked uniformity among the various programs worldwide. However, this review highlights the most consistent screening and scoring procedures that are simple to use and provide reliable results. Sources of resistance to the major necrotrophic fungi are summarized for each of the cool season food legumes. Marker-assisted selection is underway for Ascochyta blight of pea, lentil and chickpea, and Phomopsis blight of lupin. Other measures such as fungicidal control and cultural control are also reviewed. The emerging genomic information on the model legume, Medicago truncatula, which has various degrees of genetic synteny with the cool season food legumes, has promise for identification of closely linked markers for resistance genes and possibly for eventual map-based cloning of resistance genes. Durable resistance to the necrotrophic pathogens is a common goal of cool season food legume breeders.     

Resistance of Triticum dicoccoides to infection with Erysiphe graminis tritici

Summary The reactions of 233 Triticum dicoccoides acessions, collected at 10 sites in Israel and elsewhere, to infection with cultures of Erysiphe graminis tritici, were determined. The reactions indicated that the number of sources of resistance to E. graminis tritici which can be obtained from T. dicoccoides plants growing wild in Israel and elsewhere is almost unlimited. One hundred and fourteen or 49% of the accessions were resistant, and 137 or 59% of the accessions were resistant or moderately resistant to infection with four cultures of E. graminis tritici which possess the virulence genes corresponding to most of the identified resistance genes in wheat. Accessions collected at sites with marginal habitats where T. dicoccoides grows poorly and has lower grain weight, were more susceptible than were accessions collected at sites with an optimal habitat for growth of T. dicoccoides. The results agreed with those in a previous study with Hordeum spontaneum, and indicate that to obtain H. spontaneum or T. dicoccoides accessions with the highest level of resistance to the powdery mildew pathogens, plants should be collected at sites in ecological and geographic regions where those two species occupy optimum habitats and are exposed to the powdery mildew pathogens.     

Evaluation of resistance sources and inheritance of resistance in kidney bean to Indian virulences of Colletotrichum lindemuthianum

Summary Forty nine common bean lines comprising of exotic accessions and locally grown cultivars evaluated against Colletotrichum lindemuthianum exhibited differential resistance to its races in Himachal Pradesh, a north-western Himalayan state of India. Some exotic accessions like G 2333, Cornell 49242, PI 207262, Mexique 222, TO, Perry Marrow, Kaboon and Widusa were resistant to more than five Indian races, whereas two Indian accessions KRC-5 and Hans showed resistance to six and four races, respectively. However, nine accessions KRC-8, KR-40, KR-43, KR-81, KR-62-2, KR-90, KR-142, KR-148, and KR-216 were resistant to three races. Race specific resistance has been observed in different bean cultivars. Studies on inheritance of resistance in exotic accession G 2333 and Indian accession, KRC-5 showed that two independent dominant genes conferred resistance in G 2333 to race 3 and 515 and a single dominant gene controlled resistance in KRC-5 to race 775, indicating resistance from these sources is easily transferable to the locally adapted susceptible cultivars.     

The location and characteristics of novel sources of resistance to Bremia Lactucae Regel (downy mildew) in wild Lactuca L. Species

Summary Twenty-one accessions of 3 wild Lactuca species which could be hybridised with L. sativa, the cultivated lettuce, were inoculated at different stages of plant development with 3 multivirulent isolates of Bremia lactucae. Nineteen sources of resistance to B. lactucae, not attributable to the previously recognised resistance factors 1–11 were identified. Two lines of L. serriola showed similar resistance patterns as lines carrying R11. The resistance of some accession was incomplete particularly at the seedling stage and this phenomenon may be race specific.Tests on segregating F₂ populations of crosses between 2 different L. serriola accessions and L. sativa cultivars showed that the resistance in one line (LSE/18) appears to be inherited as a single dominant gene, which is sometimes incomplete in expression and allelic to either Dm6 or R7. The segregation patterns for resistance in PI 281876 did not give readily interpretable ratios.To assess the frequency of occurrence in B. lactucae populations of virulence factors to overcome this novel resistance, 11 of the novel sources of resistance were inoculated with numerous collections of the pathogen from the UK, Czechoslovakia and elsewhere and found to show a high level of resistance.     

Resistance to powdery mildew in wild emmer (Triticum dicoccoides Körn.)

Summary Wild emmer from 73 collection sites, including 107 accessions from Israel, two from Lebanon and one from Turkey, were evaluated for resistance to powdery mildew in field nurseries in Israel and the Netherlands.The wild emmer entries displayed a diversity of responses to powdery mildew infection, ranging from high resistance to complete susceptibility. Most entries were resistant in at least one of the nurseries; several entries proved to be resistant in all the tests.Comparing the reactions of 47 wild emmer accessions tested in six nurseries, 11 markedly different patterns were discerned, indicating the probable presence of several different resistance genes.Genes for resistance to powdery mildew appear to be very common in wild emmer indigenous to Israel. Resistance was found in accessions from most collection sites, in all the geographic regions represented in the collection.The common occurrence of resistance and the apparent diversity of genotypes makes wild emmer a rich gene-pool for resistance to powdery mildew. Since genes for resistance to wheat pathogens can be quite readily transferred to cultivated wheat, wild emmer may be utilized as a valuable source of powdery mildew resistance in wheat breeding.     

Relation in the inheritance of resistance to Pseudoperonospora cubensis rost and Sphaerotheca fuliginea poll. in cucumber (Cucumis sativusl. L.)

Summary Among the genes for resistance against Sphaerotheca fuliginea, the gene s for resistance of the hypocotyl is linked to or identical with gene p for resistance to Pseudoperonospora cubensis. This gene s is present in the cultivars Ashley, Poinsett, Natsufushinari, Pixie, Taipei and in accessions PI 179676. PI 212233 and PI 234517.Although gene s initially brings about some resistance of the true leaf, it appears that when s is present. other genes are necessary for complete resistance to S. fuliginea.     

Genetic linkage of QTLs for late blight resistance and foliage maturity type in six related potato progenies

A set of test crosses of diploid potatoes was used to identify QTLs for foliage resistance against Phytophthora infestans and QTLs for foliage maturity type, and to assess their genetic relationship. The most important locus for both traits was found on chromosome 5 near marker GP21: the allele of marker GP21 that is associated with resistance to late blight is also associated with late foliage maturity. An additional QTL with a small effect on foliage maturity type was identified on chromosome 3, and additional QTLs for late blight resistance were found on chromosomes 3 and 10. Another QTL was detected on chromosome 7 when resistance was adjusted for the effect of foliage maturity type. The additional QTLs for resistance against P. infestans on chromosomes 3 and 10 seem to be independent of foliage maturity type and are not affected by epistatic effects of the locus on chromosome 5. The effects of the additional QTLs for resistance are small, but early maturing genotypes that necessarily have the allele for susceptibility for late blight on chromosome 5 may benefit from the resistance that is provided by these QTLs on chromosomes 3 and 10.     

A rapid screening technique to combine resistance to halo blight and bean common mosaic virus in Phaseolus vulgaris L.

Summary Seven bean lines (Phaseolus vulgaris L.) with differential resistance or susceptibility to race 2 of halo blight (Pseudomonas syringae pv. phaseolicola) and a necrosis-inducing isolate of bean common mosaic virus were inoculated with one or both pathogens in combination, to determine the feasibility of dual screening to identify resistance to both pathogens simultaneously. Dual screening yielded the same results as separate screenings. Neither pathogen affected the disease expression of the other. Simultaneously screening for resistance to both pathogens will shorten the recurrent screening-selection cycle of hybridization programs, and accelerate development of resistant cultivars.Abbreviations BCMV Bean Common Mosaic Virus - cvs Cultivars - HB Halo blight - Inoc. Pt. Inoculation point - NLL Necrotic local lesion - React Reaction - SVN Spreading veinal necrosis, System chloro-Systemic chlorosis - VN Vascular necrosis     

Leaf rust and stripe rust resistance genes transferred to common wheat from Triticum dicoccoides

Linked leaf rust and stripe rust resistance genes introduced from Triticum dicoccoides protected common wheat seedlings against a range of pathotypes of the respective pathogens. The genes were chromosomally mapped using monosomic and telosomic analyses, C-banding and RFLPs. The data indicated that an introgressed region is located on wheat chromosome arm 6BS. The introgressed region did not pair with the ‘Chinese Spring’ 6BS arm during meiosis possibly as a result of reduced homology, but appeared to pair with 6BS of W84-17 (57% of pollen mother cells) and ‘Avocet S’. The introgressed region had a very strong preferential pollen transmission (0.96–0.98) whereas its transmission through egg cells (0.41–0.66) varied with the genetic background of the heterozygote. Homozygous resistant plants had a normal phenotype, were fertile and produced plump seeds. Symbols Lr53 and Yr35 are proposed to designate the respective genes.     

Disease resistance in protected crops and mushrooms

Summary Cultivars of tomatoes, cucumbers, lettuce and peppers have been bred for resistance to one or more pathogens. Some tomato and cucumber cultivars have resistance to a wide range of diseases. Resistance has been transient in many cases and a succession of cultivars with new genes or new combinations of resistance genes has been necessary to maintain control. There has been a number of notable exceptions and these have included durable resistance to such pathogens asFulvia fulva and tomato mosaic virus. With lettuce the resistance situation is complicated by the occurrence of fungicide resistant pathotypes. There are no strains ofAgaricus bisporus purposely bred for disease resistance.In protected flower crops only resistance to Fusarium wilt in carnations has been purposely bred but differences in disease resistance are apparent in cultivars of many ornamental crops. This is particularly so in chrysanthemums where there are cultivars with resistance to many of the major pathogens. Similar situations occur with other flower crops and pot plants. Cultivars of some species have not been systematically investigated for resistance.The need for genetic resistance will increase with the further reduction, in the limits on pesticide use and an increasing public awareness and importance of pesticide pollution.ADAS is an executive agency of the Ministry of Agiculture, Fisheries and Food and the Welsh Office.     

Search for novel genes for resistance to powdery mildew (Sphaerotheca fuliginae) in cucumber (Cucumis sativus)

Summary The current powdery mildew (Sphaerotheca fuligninea) resistant cucumber varieties suffer from leaf chlorosis during autumn, winter and early spring cultivation in the Netherlands. Therefore screening was carried out for novel powdery mildew resistance genes. From 177 accessions, derived from different sources, 108 accessions proved to be partially resistant to S. fuliginea. Crosses were made with 53 resistant accessions to distinguish the presence of novel genes. It is likely that the accessions C. sativus 2145, C. sativus LV 41, PI 188807, Vladivostokij, White and Yellow 1 have one or more recessive powdery mildew resistance genes, different from powdery mildew resistance genes of the line NPI, which was used for variety breeding. Powdery mildew resistance tests with S. fuliginea give similar results in different regions of the world.Abbreviations pmr powdery mildew resistance     

Induced mutagenesis for the development of multiline cultivars in bread wheat

Summary Variation for resistance toPuccinia graminis f.sp.tritici, P. recondita f.sp.tritici andP. striiformis was induced in theTriticum aestivum cultivar Lalbahadur using nitrosomethyl urea. Variations were isolated from the M₂ population in the post-seedling stage in the field when infected with a mixture of races of each of the three rusts. Plants exhibiting simultaneous resistance to stem rust, leaf rust and yellow rust were indentified. Repeated screening in the subsequent generations confirmed the resistance of the mutant lines that are morphologically similar to the parental cultivar. The rust resistance of 20 mutant lines was also confirmed at the seedling stage using individual races of stem rust and leaf rust. The different patterns observed in the mutant lines tested against a wide range of races show that these lines can be used as components of a multiline. The patterns of variation compared with those of the known genes for resistance against the Indian races of the pathogens suggest that the mutations for rust resistance are due to factor different from those already known in bread wheat, providing a broadened genetic base for future breeding programmes.     

Molecular markers and allelic relationships of anthracnose resistance gene cluster B4 in common bean

Angular leaf spot is one of the major diseases of the common bean. The extensive genetic variability of this pathogen requires the constant development of new resistant cultivars. Different sources of resistance have been identified and characterized. For the State of Minas Gerais, Brazil, four main resistance sources were found: Mexico 54, AND 277, MAR 2 and Cornell 49-242. Independent characterization of these genotypes demonstrates that resistance in all four sources is dominant and monogenic. However, there are no studies on the relationship and independence of these genes. In the present work, allelism tests were carried out to understand the relationship among the resistance genes present in these four resistance sources. The data revealed a much higher complexity in the resistance inheritance of these genes than previously reported. It was demonstrated that Cornell 49-242 possesses a dominant gene (Phg-3); Mexico 54 possesses three genes, denominated Phg-2, Phg-5 and Phg-6. In MAR 2, two genes were found, one independent designated Phg-4 and the other, an allelic form of Phg-5, denominated of Phg-5². Allelic forms were also found in AND 277, Phg-2², Phg-3² and Phg-4². These results have special importance for breeding programs aiming to pyramid resistance genes.     

Screening of Portuguese cole landraces (Brassica oleracea L.) with Peronospora parasitica and Plasmodiophora brassicae

Summary Forty-four landraces of Portuguese coles (Brassica oleracea L.), representing different morphotypes, were screened at the cotyledon stage for downy mildew resistance, caused by Peronospora parasitica. Two isolates of the pathogen, PHW 630 and PHW 828, were used in the experiment. The mean disease interaction phenotype varied among the landraces. Overall, the Portuguese coles have shown resistance to both isolates, particularly to isolate PHW 828. Some landraces were identified with differential reaction to the two P. parasitica isolates. Some of the Portuguese coles are, therefore, good sources of resistance genes to P. parasitica, and could be used in breeding programs or in the development of a downy mildew host differential set. Among the morphotypes tested, Couve Algarvia (ISA 207), Couve Galega (ISA 243 and 187) and Couve Glória de Portugal (ISA 84) have presented the highest resistance interaction. For the development of host differential sets, the landraces ISA 2 (Penca de Mirandela), ISA 35 (Couve de Valhascos) and ISA 600 (Penca Espanhola) could be further selected for the differentiation of P. parasitica isolates. They are resistant to PHW 828 and susceptible to PHW 643.The same morphotypes and landraces were also screened at the seedling stage for resistance to clubroot, caused by Plasmodiophora brassicae Woron.. Although some variation for resistance could be noticed in some the landraces tested, mainly Galega kale types, none of them presented a sufficient level of resistance as to be directly be useful in breeding for resistance to race 6 of P. brassicae.     

Screening techniques and sources of resistance to foliar diseases caused by fungi and bacteria in cool season food legumes

Screening techniques are an important component of the overall strategy of breeding for resistance to diseases in cool season food legumes. Suitable screening methods have been developed for several major foliar diseases of chickpea, pea, faba bean, and lentil, and sources of resistance have been identified. International cooperation plays an important role in promoting research and keeping collections of cultivated species and their wild relatives. New biotechnological approaches are promising for enhancing the practical use of genes for resistance.     

Development of common bean (Phaseolus vulgaris L.) lines resistant to the bean pod weevil,Apion godmani Wagner,in Central America

Summary The larva of the bean pod weevil (BPW), Apion godmani Wagner (Coleoptera: Curculionidae), causes serious yield losses in common bean (Phaseolus vulgaris L.) in Mexico and Central America, by consuming the seed as it develops in the immature pod. Resistance to the BPW was identified in bean germplasm of highland Mexican origin, and these sources of resistance were incorporated into a pedigree breeding program to recover locally adapted lines resistant to Bean Common Mosaic Virus and BPW, with commercial grain for Guatemala, Honduras and El Salvador. These lines yielded as well as or better than local cultivars in the absence of the insect, and better than local cultivars when the BPW was present. Resistance appeared to be governed by several genes, and was stable across geographic areas, seasons and planting systems.     

兼抗全蚀病和白粉病小麦新种质的创制与鉴定

TaLTP5是从小麦中分离到的一个脂质转移蛋白编码基因。利用基因枪介导法将TaLTP5表达载体pA25-TaLTP5转入抗白粉病的小麦品种扬麦18 (含抗白粉病基因Pm21)中, 旨在选育兼抗全蚀病和白粉病的小麦新种质。对转基因小麦T₀~T₃代植株中引入TaLTP5基因进行分子检测和抗病性鉴定。PCR检测、Southern杂交分析结果表明, 外源TaLTP5基因已转入、整合到3个转基因小麦株系的基因组中, 并能稳定遗传; 荧光定量RT-PCR的分析以及全蚀病菌的接种与鉴定结果表明, 与受体小麦扬麦18相比, 这3个转基因小麦株系中TaLTP5表达量显著提高, 其对全蚀病的抗性也明显增强。对3个转基因株系的Pm21分子标记和白粉病抗性鉴定表明, 外源TaLTP5基因的导入没有影响受体小麦对白粉病抗性, 说明这些转基因株系为兼抗全蚀病和白粉病小麦新种质。