蚕豆和豌豆锈病病原菌的分子鉴定

为明确我国热带和亚热带地区蚕豆Vicia faba和豌豆Pisum sativum锈病的病原菌种类,通过致病性测定和ITS序列系统发育分析对来自我国云南省玉溪市的4份豌豆锈菌分离物及云南、广西、重庆和四川省(区、市)的5份蚕豆锈菌分离物进行系统鉴定。结果显示,分离自豌豆的锈菌WX1分离物对蚕豆和豌豆均具有高致病性,在侵染叶片上产生大量锈子器;分离自蚕豆的锈菌CX3分离物仅对蚕豆具有高致病性,能在叶片上产生大量夏孢子,而对豌豆的致病性相对较低,仅产生少量的夏孢子堆;分离物WX1和CX3对小扁豆和鹰嘴豆不具有致病性。基于ITS序列系统发育分析表明,所有不同寄主来源的蚕豆单胞锈菌分离物均聚类于一个系统发育组,但分离自蚕豆和豌豆的分离物分别聚类在不同的亚组。表明分离自云南省玉溪市豌豆上的蚕豆单胞锈菌Uromyces viciae-fabae应为豌豆专化型,定名为U. viciae-fabae ex P. sativaum,而来源于云南、广西、重庆和四川省(区、市)的蚕豆锈病病原菌为蚕豆专化型U. viciae-fabae ex V. faba。     

Australian Uromyces viciae-fabae: Host and nonhost interaction among cultivated grain legumes

Uromyces viciae-fabae, rust of faba bean, parasitizes other legume crops such as lentils (Lens culinaris) and field peas (Pisum sativum) in some environments. In this study we examined the host range of two Australian isolates of U. viciae-fabae collected and purified from a faba bean crop and classified as U. viciae-fabae ex V. faba. Field pea (P. sativum), chickpea (Cicer arientinum), lupin (Lupinus spp.), lentil (L. culinaris), and mung bean (Vigna radiata) genotypes were tested with these isolates, as well as resistant and susceptible genotypes of the faba bean host. Race specificity for these two pathogen isolates was observed on Vicia faba, with two faba bean genotypes showing partial resistance. Both U. viciae-fabae isolates also colonized field pea seedlings and successfully produced uredinia under glasshouse conditions, despite this fungus not being known as a pathogen of Australian field pea crops. No sporulation of either isolate of U. viciae-fabae ex V. faba was observed on any of the remaining legume species tested. However, obvious differences in fungal growth were observed, ranging from small infection sites with very rare haustorium formation in mung bean to more extensive growth and the development of potential uredinial structures in chickpea. These observations are discussed in relation to the phylogenetic relationship of these host and nonhost species.     

Pea seed-borne mosaic virus: occurrence in faba bean (Vicia faba) and lentil (Lens culinaris) in West Asia and North Africa,and further information on host range,transmission characteristics,and purification

In a survey for viruses of cultivated legumes in West Asia and North Africa, pea seed-borne mosaic virus (PSbMV) was found in faba bean, lentil and pea. Using ELISA, it was detected in 107 out of 1554 faba bean samples and 40 out of 496 lentil samples with virus-like symptoms collected in Algeria, Egypt, Ethiopia, Jordan, Lebanon., Libya, Morocco, Sudan, Tunisia and Turkey.A pea isolate (SP9-88) from Syria was further characterized. Out of 57 plant species tested, 35 were found susceptible, 19 of which are newly reported hosts of the virus. The virus was transmitted efficiently in the non-persistent manner by five aphid species, especiallyMyzus persicae. Purification from systemically infected faba bean plants yielded 10–15 mg of purified virus per kg of infected tissue. Sap-inoculation of the food and forage legume species chickpea, faba bean, lentil, pea,Vicia narbonensis, V. sativa, Lathyrus ochrus andL. sativus at flowering stage led to 66.0, 40.5, 44.6, 49.2, 31.7, 7.5, 35.7 and 12.0% yield loss, respectively, and to seed-transmission, rates of 0.7, 6.0, 10.8, 1.1, 0.3, 0.2 and 0.4%, respectively. No transmission was detected in chickpea seed embryo axes. However, the virus was detected in the seed coat of SPbMV-infected chickpea at an estimated rate of 1.81%.     

Reaction of genotypes from several species of grain and forage legumes to infection with a French pea isolate of the oomycete <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis>

The susceptibility/resistance to Aphanomyces euteiches of various genotypes (cultivars and breeding lines) of several grain legume species was assessed in controlled conditions. A total of 279 genotypes from the major grain legumes grown in temperate climates (faba bean, chickpea, lentil, lupin and common vetch) and three other legumes frequently cultivated in France (French bean, clover and alfalfa) were screened with one pea-infecting isolate from France. Four different categories of susceptibility/resistance were identified among the legume species/cultivars tested with the pea A. euteiches isolate: (1) susceptible legume species (lentil, alfalfa, French bean) among which low levels of partial resistance was observed; (2) legume species including susceptible genotypes and genotypes with high levels of resistance (common vetch, faba bean and clover), (3) species with a very high level of resistance (chickpea) and (4) species displaying no symptoms (lupin). It is therefore important to consider pathogen-species and pathogen-genotype interactions when defining the host specificity of A. euteiches and considering the possible role of different legume species in increasing or decreasing the soil inoculum potential.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

Broad bean mottle virus in Morocco; curculionid vectors,and natural occurrence in food legumes other than faba bean (Vicia faba)

Broad bean mottle virus (BBMV) was transmitted from infected to healthy faba-bean plants by the curculionid weevilsApion radiolus Kirby,Hypera variabilis Herbst,Pachytychius strumarius Gyll,Smicronyx cyaneus Gyll, andSitona lineatus L. The latter appeared to be an efficient vector: acquisition and inoculation occurred at the first bite, the rate of transmission was c. 41%, and virus retention lasted for at least seven days.S. lineatus transmitted the virus from faba bean to lentil and pea, but not to the three genotypes of chickpea tested. This is the first report on the generaHypera, Pachytychius, andSmicronyx as virus vectors, and onA. radiolus, H. variabilis, P. strumarius, andS. cyaneus as vectors of BBMV.Out of 351 samples of food legumes with symptoms suggestive of virus infection, 16, 11, 19, and 17% of the samples of chickpea, lentil, pea, and common bean, respectively, were found infected when tested for BBMV in DAS-ELISA. This is the first report on the natural occurrence of BBMV in chickpea, lentil, pea, and common bean. The virus should be regarded as a food-legume virus rather than a faba-bean virus solely, and is considered an actual threat to food legume improvement programmes.     

Molecular identification of 16SrII-D subgroup phytoplasmas associated with chickpea and faba bean in Sudan

In January 2011, symptomatic chickpea and faba bean plants were observed in fields located in the Gezira state (Sudan). Faba bean plants showed yellowing and stunting, whereas chickpea plants presented yellowing, reddening and little leaves. The disease etiology was investigated using nested polymerase chain reaction (PCR) with phytoplasma-specific primers which amplify a fragment of the 16S rRNA gene. Sequencing and restriction fragment length polymorphism (RFLP) analyses revealed that the tested phytoplasmas belonged to the group 16SrII. Phylogenetic analyses of the 16S rRNA gene of the obtained sequences indicated that the chickpea and faba bean phytoplasmas from Sudan were more closely related to the phytoplasmas subgroup 16SrII-D. To our knowledge, this is the first report of phytoplasmas from the group 16SrII-D infecting chickpea in Sudan, and faba bean worldwide.     

Broad bean mottle virus in Morocco; variability,interaction with food legume species,and seed transmission in faba bean,pea, and chickpea

Biological indexing of faba-bean samples collected during an earlier virus survey in Morocco revealed variation in symptom severity among isolates of broad bean mottle virus (BBMV). When seven selected isolates from Morocco and three from Algeria, Sudan, and Tunisia were further compared, they could be divided into mild, severe, and intermediate isolates, according to their pathogenicity on a number of food-legume genotypes tested. The Moroccan isolate SN1 and the Sudanese SuV256 were very mild, and deviant also in their effect onGomphrena globosa, whereas the Tunisian TV75-85 and the Moroccan VN5 were virulent. Representative isolates were indistinguishable, however, in coat-protein molecular weights, and they reacted similarly to the antisera to a Moroccan and a Syrian isolate in electro-blot immunoassay.Promising ICARDA breeding lines and accessions—ten each of pea and lentil, nine of chickpea, and twelve of faba bean-were all found vulnerable (susceptible and sensitive) to all isolates. Within each food-legume species, vulnerability varied from high to moderate, and no immunity was detected. Virus concentrations in faba-bean lines suggest that isolates differ in virulence rather than in aggressiveness, and that the differences in vulnerability among the lines are due to differences in sensitivity rather than in susceptibility.When pooled seed samples were germinated and seedlings were tested for BBMV in DASELISA, the virus was found seed-transmitted in faba bean, chickpea, and pea at transmission rates of ca 1.2, 0.9, and 0.1%, respectively. This is the first report on seed transmission of BBMV in faba bean, when occurring on its own, and the first record of such seed transmission in chickpea and pea.     

Nematodes of food legumes in the Mediterranean Basin1

Several endoparasitic nematodes have been reported on leguminous plants in the Mediterranean area. The most widespread are the root-lesion nematodes Pratylenchus mediterraneus, P. penetrans and P. thornei. Symptoms induced by these nematodes usually are not very impressive, but 50% yield loss of chickpea may occur. Among root-knot nematodes, Meloidogyne artiellia is associated with severe yield losses of chickpea in Italy, Spain and especially Syria. Tolerance limits of 0.14 and 0.02 of this nematode per ml soil are reported for winter and spring-sown chickpea, respectively. Meloidogyne incognita and M. javanica can be noxious to French bean and cowpea in sandy soil. The cyst nematode Heterodera goettingiana reduces yields of pea, broad bean and vetch when its population densities exceed 0.5, 1, and 2.1 eggs per g of soil, respectively. Heterodera ciceri occurs in northern Syria and Turkey and is responsible for economic yield losses of chickpea and lentil in fields infested with more than 1 or 2.5 eggs per g of soil, respectively. Pea and grass pea also suffer from infestation of this nematode. The stem and bulb nematode Ditylenchus dipsaci causes severe decline of broad bean, pea and probably lentil during wet seasons. Other nematodes, although present in moderate numbers, appear to have little importance.     

Towards identifying pathogenic determinants of the chickpea pathogen <Emphasis Type="Italic">Ascochyta rabiei</Emphasis>

Ascochyta blight is a serious disease of cool-season grain legumes (chickpea, faba bean, lentil and pea) caused by fungal species of the anamorphic genus Ascochyta and related genera. Despite extensive studies on the biology, ecology, epidemiology and management of the disease, little is known about the pathogenic determinants of these pathogens. This research aims at using Ascochyta rabiei as a model for the genus in investigating genetic factors of pathogenicity, with the ultimate goal of elucidating pathogenic mechanisms. Three advances were made: (1) insertional mutants with altered pathogenicity were identified through in vivo screening, and genomic regions adjacent to the insertion sites in selected mutants were determined; (2) a phage library of A. rabiei genomic DNA was constructed, and the library was estimated to provide complete coverage of the A. rabiei genome. This library was used successfully to recover clones with DNA adjacent to insertional mutation sites and to isolate specific genes; (3) DNA probes specific for an acyl-CoA ligase (cps1) and a polyketide synthase gene (pks1) were developed and library clones containing the corresponding genomic regions were identified from the phage library. These advances provide the foundation and necessary tools for experimentation of ectopic complementation assays and targeted mutagenesis to elucidate the genetic mechanisms of pathogenicity of A. rabiei.     

番茄黄化曲叶病毒在湖北首次报道

2014年春季,在湖北省武汉市发现种植的番茄表现植株矮缩,叶片上卷,叶缘黄化等症状。PCR检测结果显示,所有采集的病样中均存在菜豆金色花叶病毒属病毒。进一步通过滚环扩增方法获得了该病毒的湖北分离物HB01的全基因组。基因克隆及序列分析结果表明,该病毒基因组全长为2 781nt,与已报道的番茄黄化曲叶病毒(TYLCV)各分离物同源性在89.0%以上,而与来自中国不同地区的TYLCV分离物的同源率均在97.0%以上。因此,HB01属于TYLCV的一个分离物。     

A mild strain of broad bean mottle virus from faba bean (Vicia faba L.) in the Sudan

A new strain of broad bean mottle virus, isolated from faba bean (Vicia faba L.) in the Sudan, is described. It differs considerably from known isolates by its nearly symptomless infection of faba bean in spite of high concentrations of the virus in infected plants. It does not differ from regular isolates in gel-diffusion serology, light and electron microscopy, host range and symptoms in major hosts other than faba bean. It may constitute a potential threat to other food legumes in the region.     

Broad bean mottle virus: Identification,host range,serology, and occurrence on faba bean (Vicia faba) in West Asia and North Africa

One of the faba bean viruses found in West Asia and North Africa was identified as broad bean mottle virus (BBMV) by host reactions, particle morphology and size, serology, and granular, often vesiculated cytoplasmic inclusions. Detailed research on four isolates, one each from Morocco, Tunisia, Sudan and Syria, provided new information on the virus.The isolates, though indistinguishable in ELISA or gel-diffusion tests, differed slightly in host range and symptoms. Twenty-one species (12 legumes and 9 non-legumes) out of 27 tested were systemically infected, and 14 of these by all four isolates. Infection in several species was symptomless, but major legumes such as chickpea, lentil and especially pea, suffered severely from infection. All 23 genotypes of faba bean, 2 of chickpea, 4 of lentil, 11 out of 21 ofPhaseolus bean, and 16 out of 17 of pea were systemically sensitive to the virus. Twelve plant species were found to be new potential hosts and cucumber a new local-lesion test plant of the virus.BBMV particles occurred in faba bean plants in very high concentrations and seed transmission in this species (1.37%) was confirmed.An isolate from Syria was purified and two antisera were produced, one of which was used in ELISA to detect BBMV in faba bean field samples. Two hundred and three out of the 789 samples with symptoms suggestive of virus infection collected in 1985, 1986 and 1987, were found infected with BBMV: 4 out of 70 (4/70) tested samples from Egypt, 0/44 from Lebanon, 1/15 from Morocco, 46/254 from Sudan, 72/269 from Syria and 80/137 from Tunisia. This is the first report on its occurrence in Egypt, Syria and Tunisia. The virus is a potential threat to crop improvement in the region.Samenvatting Eén van de in West-Azië en Noord-Afrika in faba-boon aangetroffen virussen werd geïdentificeerd als het tuinbonevlekkenvirus (broad bean mottle virus) op grond van waardplantreacties, deeltjesvorm en-grootte, serologische eigenschappen en granulaire, vaak gevacuoliseerde celinsluitsels. Verder onderzoek aan vier isolaten uit respectievelijk Marokko, Tunesië, Soedan en Syrië verschafte nieuwe informatie, over het virus.De in ELISA of gel-diffusietoetsen serologisch niet te onderscheiden isolaten verschilden enigszins in waardplantenreeks en symptomen. Van 27 getoetste plantesoorten werden 21 systemisch geïnfecteerd (12 vlinderbloemigen, en 9 niet-vlinderbloemigen) waarvan 14 door alle vier isolaten. In vele ervan was de infectie symptoomloos, maar belangrijke als gewas geteelde vlinderbloemigen, zoals erwt, linzen en kekererwt, leden ernstig onder aantasting. Alle 23 getoetste faba-boongenotypen, beide van kekererwt, alle vier van linzen, 11 van de 21 getoetste vanPhaseolus-boon en 16 van de 17 van erwt bleken systemisch gevoelig voor het virus. Twaalf plantesoorten, bleken nieuwe potentiële waardplanten en komkommer een nieuwe lokale-lesietoetsplant voor het virus te zijn.In faba-boneplanten kwam, het virus in hoge concentratie voor en overdracht met zaad (1.37%) in deze soort kon worden bevestigd.Een Syrisch isolaat werd gezuiverd en twee antisera werden bereid, waarvan één werd gebruikt voor de detectie van het virus in te velde verzamelde monsters. Van 789 in 1985 tot en met 1987 verzamelde bladmonsters, met symptomen die deden denken aan virusinfectie, bleken 203 het virus te bevatten en wel 4 van de 70 (4/70) uit Egypte, 0/44 uit Libanon, 1/15 uit Marokko, 46/254 uit Soedan, 72/269 uit Syrië en 80/137 uit Tunesië. Het virus was nog niet eerder aangetoond in Egypte, Syrië en Tunesië.De grote verbreiding, grote kunstmatige waardplantenreeks, overdracht met zaad, en pathogeniteit voor een aantal belangrijke vlinderbloemige gewassen maken het virus tot een potentiële bedreiging van de programma's tot verbetering van de teelt van de bedoelde gewassen in het betrokken gebied.     

Cryptic diversity,multilocus phylogeny,and pathogenicity of cercosporoid fungi associated with common bean and cowpea

In recent years, common bean (Phaseolus vulgaris) and cowpea (Vigna unguiculata) plants in the north of Iran have exhibited symptoms resembling Cercospora leaf spot (CLS) disease. This study was initiated to elucidate the taxonomy and pathogenicity of cercosporoid taxa associated with leaf spot diseases of these two legume crops in Iran. A total of 138 samples with CLS symptoms were collected from cultivated common bean and cowpea species in northern Iran and subjected to microscopic examination, resulting in identification of 98 Cercospora and 59 Pseudocercospora samples. A six-locus phylogenetic analysis (ITS, actA, tef1, gapdh, his3, and cmdA) coupled with examination of the morphology of 42 representative isolates from these samples confirmed that several cercosporoid fungi occur on common bean and cowpea in Iran. Five Cercospora species (C. iranica, C. cf. flagellaris, Cercospora sp. G, Cercospora sp. T, and C. vignigena) and two Pseudocercospora species (P. griseola f. griseola and P. cf. cruenta) were found; of these, C. cf. flagellaris was the dominant species, occurring on both common bean and cowpea. Pathogenicity tests confirmed that all seven species could infect leaves of common bean and/or cowpea. This is the first report of C. iranica, Cercospora sp. G, and Cercospora sp. T associated with common bean and/or cowpea in the world. In addition, C. vignigena was recorded for the first time in Iran. Results achieved in this study will assist strategies for the management of CLS disease of common bean and cowpea.     

Screening faba bean for chocolate spot resistance: evaluation methods and effects of age of host tissue and temperature

Chocolate spot is an important disease of faba bean (Vicia faba) caused by the necrotrophic fungus Botrytis fabae. The aims of this work were: i) to compare different methods of screening for resistance; ii) to assess the influence of the age of host tissue and temperature on this pathosystem. To this effect, a collection of 42 faba bean accessions was evaluated in mature plant stage in the field in Cordoba (Southern Spain) and in detached leaflet and whole plant tests under controlled conditions. Field results correlated better with those of the whole plant test than with those of the detached leaflet assay. Integration of results from the field and whole plant experiments resulted in the selection of six accessions of interest as sources of resistance. Influence of leaf age on disease development was found to be genotype dependent. Older leaves were more susceptible than younger ones in 23 accessions, while no difference between leaf ages was detected in the remaining accessions. The effects of plant age and temperature were assessed by a whole plant test on seven accessions at two plant ages (4 and 7 weeks) and three temperatures (13, 20, and 25°C). Results showed that the differential genotypic responses to B. fabae were not significantly influenced by either plant age or temperature, although there was a tendency towards lower susceptibility to chocolate spot in faba bean plants as they become older. Further, a partial high-temperature, young-plant resistance was detected.     

Agroinoculation Shows Tobacco leaf curl Yunnan virus is a Monopartite Begomovirus

We demonstrated that only 2 out of 15 isolates of Tobacco leaf curl Yunnan virus (TbLCYNV) were associated with the satellite DNAβ molecules. To investigate the infectivity of this virus, an infectious clone of TbLCYNV isolate Y143 (TbLCYNV-Y143) was agroinoculated or whitefly transmitted into Nicotiana benthamiana, N. glutinasa, Petunia hybrida and N. tabacum. TbLCYNV-Y143 alone was able to induce severe upward leaf curling, vein thickening or stunt symptoms in these plants. Co-inoculation of TbLCYNV-Y143 with DNAβ molecules associated with other begomoviruses induced similar symptom types on these plants. This indicates that TbLCYNV is a monopartite begomovirus. The relevance of results that only two isolates of TbLCYNV were associated with DNAβ molecules is discussed.     

Mitigation of vanillic acid-promoted faba bean Fusarium wilt by faba bean–wheat intercropping

Long-term continuous monocropping of faba beans increases the incidence of faba bean wilt, while faba bean–wheat intercropping can effectively control it. This study aimed to understand the underlying mechanism of faba bean–wheat intercropping for the control of Fusarium oxysporum and vanillic acid (VA)-promoted occurrence of faba bean wilt. The occurrence of faba bean wilt was investigated among the monocropped and intercropped plants of faba beans in a field experiment. The contents and types of phenolic acids were examined in the rhizosphere soil. Monocropped and intercropped faba beans were examined under the dual stress of F. oxysporum and different concentrations of VA (0, 50, 100, 200 mg/L) to understand the alleviating mechanism of faba bean–wheat intercropping. Exogenous addition of high concentrations of VA significantly inhibited the growth and reproduction of F. oxysporum, but under the dual stress of F. oxysporum and different concentrations of VA, it significantly inhibited the defence enzymes of faba bean roots, stems, and leaves, and rhizosphere soil enzymes. Interestingly, faba bean–wheat intercropping alleviated VA stress and thereby the incidence and disease index of faba bean Fusarium wilt by improving plant resistance and soil enzyme activity. The dual stress of F. oxysporum and VA promotes the occurrence of Fusarium wilt by damaging the defence system of the faba bean root system and rhizosphere soil environment. However, faba bean–wheat intercropping effectively alleviates the autotoxicity of VA by improving the physiological and biochemical resistance of faba beans and soil enzyme activities, and thus controls the occurrence of Fusarium wilt.     

Leaf Spot Disease of Broad Bean (vicia faba L.) Caused by Alternaria tenuissima— A New Disease in Japan

Farmers' fields were surveyed for a new disease on broad bean in different parts of Shimane Prefecture. The disease was found in all surveyed fields. The fungus frequently associated with diseased broad bean leaves having the characteristic leaf spot symptom was identified as Alternaria tenuissima, based on morphological characteristics. The initial lesion was brown, water soaked, circular to slightly irregular. Then the lesion enlarged and became concentric. Mature leaves had coalescing necrosis surrounded by yellowing. Older leaves of the plant were particularly affected. In a later stage of the disease, the leaves became blighted from the margin to the center and most of the diseased plants defoliated. In severely infected fields, circular to irregularly shaped lesions also appeared on stems and pods, and plants defoliated completely. All tested isolates were pathogenic to broad bean leaves. Different isolates of the fungus varied in pathogenicity on different broad bean cultivars. In a comparison of symptoms of the disease and morphological characteristics of the isolated fungus with other broad bean diseases, this disease represented a new, as yet undescribed disease of broad bean in Japan. Therefore, we propose the name "leaf spot of broad bean" and "Soramame hanten-by?" in Japanese. Received 12 April 2001/ Accepted in revised form 18 October 2001