Simultaneous temporal progress of sorghum anthracnose and leaf blight in crop mixtures with disparate patterns

Field studies were conducted at Alupe in western Kenya in 1995 and 1996 to evaluate the efficacy of crop and species mixtures for the management of sorghum anthracnose (caused by Colletotrichum sublineolum) and leaf blight (caused by Exserohilum turcicum). The progress of these diseases developing simultaneously on a susceptible sorghum cultivar planted in inter- or intra-row mixtures of varying proportions with either maize or resistant sorghum was monitored. The effects of host type and mixture patterns on disease progress were compared by parameter estimates derived from fitted Lotka-Volterra competition equations and nonlinear logistic models. Competition coefficients were not significant and their confidence intervals included zero in most cases, suggesting that interactions between C. sublineolum and E. turcicum did not occur. Mixtures of the susceptible sorghum with either the nonhost maize or the resistant sorghum delayed the time when disease is first observed and reduced the rate of disease progress and carrying capacity for both anthracnose and leaf blight, with a more pronounced effect on the latter disease. The lower efficacy of mixtures in reducing anthracnose was attributed to an aggregated spatial pattern, coupled with higher rates of progress for this disease. Intra-row mixtures were more efficient than inter-row mixtures in reducing disease development in all years. The implications of these observations for the management of sorghum diseases under small-scale farming systems are discussed.     

Genetic differentiation in populations of Exserohilum turcicum from maize and sorghum in South Africa

Exserohilum turcicum is the causal agent of northern leaf blight, a devastating foliar disease of maize and sorghum. Specificity of E. turcicum to either maize or sorghum has been observed previously, but molecular evidence supporting host specialization is lacking. The aim of this study was to compare the genetic structure of E. turcicum isolates collected from adjacent maize and sorghum fields in Delmas and Greytown in South Africa. In addition, the mode of reproduction of this pathogen was investigated. Isolates from maize (N = 62) and sorghum (N = 64) were screened with 12 microsatellite markers as well as a multiplex mating type PCR assay. No shared haplotypes were observed between isolates from different hosts, although shared haplotypes were detected between isolates from maize from Delmas and Greytown. Population structure and principal coordinate analyses revealed genetic differentiation between E. turcicum isolates from maize and sorghum. Analysis of molecular variance indicated higher among‐population variation when comparing populations from different hosts, than comparing populations from different locations. Lack of shared haplotypes, high proportion of private alleles, greater among‐population variance between hosts than locations and significant pairwise population differentiation indicates genetic separation between isolates from maize and sorghum. The high haplotypic diversity in combination with unequal mating type ratios and significant linkage equilibrium indicates that both sexual and asexual reproduction contributes to the population genetic structure of E. turcicum in South Africa.     

Inheritance of resistance in <Emphasis Type="Italic">Solanum nigrum</Emphasis> to <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Solanum nigrum, black nightshade, is a wild non-tuber bearing hexaploid species with a high level of resistance to Phytophthora infestans (Colon et al. 1993), the causal agent of potato late blight, the most devastating disease in potato production. However, the genetic mode of resistance in S. nigrum is still poorly understood. In the present study, two S. nigrum accessions, 984750019 (N19) and #13, resistant (R) and susceptible (S), respectively, to three different isolates of P. infestans, were sexually crossed. The various kinds of progeny including F1, F2, F3, and backcross populations (BC₁; F1 × S), as well as two populations produced by self-pollinating the R parent and S parent, were each screened for susceptibility to P. infestans isolate MP 324 using detached leaf assays. Fifty seedling plant individuals of the F1 progeny were each resistant to this specific isolate, similarly to the seedling plants resulting from self-pollination of the resistant R parent. Thirty seedling plants obtained from self-pollination of the S parent were susceptible. Among a total of 180 F2 plants, the segregation ratio between resistant and susceptible plants was approximately 3: 1. Among the 66 seedling plants of the BC₁ progeny originating from crossing an F1 plant with the susceptible S parent, there were 26 susceptible and 40 resistant plants to P. infestans. The segregation patterns obtained indicated monogenic dominant inheritance of resistance to P. infestans isolate MP 324 in S. nigrum acc. 984750019. This gene, conferring resistance to P. infestans, may be useful for the transformation of potato cultivars susceptible to late blight.     

Host–pathogen interaction between Phytophthora infestans and Solanum nigrum, S. villosum, and S. scabrum

Potato and tomato are the two major hosts for Phytophthora infestans causing late blight. The susceptibility of leaves and whole plants of Solanum nigrum, S. villosum, and S. scabrum to infection by P. infestans was tested under laboratory conditions. Out of 39 plants representing 38 different S. nigrum accessions, 16 were highly resistant (seven accessions did not show any symptoms of infection, nine were highly resistant showing necrotic lesions in the place of infection), and 23 plants of S. nigrum were colonized by, at least, 1 of the 2 isolates of P. infestans (17 accessions were infected with two P. infestans isolates, and 6 accessions showed different reactions depending on the isolate used for inoculation). Three accessions of S. villosum, and one accession of S. scabrum were tested and did not show any symptoms of infection. The majority of S. nigrum accessions infected by P. infestans in a detached leaf assay were also infected in the whole plant assay. The reaction of field- and greenhouse-grown plants to inoculation with P. infestans in detached leaf assays was similar, but in some cases leaves from field-grown plants reacted as resistant in comparison with the leaves from greenhouse-grown plants, which were susceptible.     

Histological characterization of the early-stage infection events of Setosphaeria turcica in maize

Northern corn leaf blight (NCLB) caused by Setosphaeria turcica is a major foliar disease of maize. The early-stage infection events of this pathogen on maize leaves are unclear. We investigated the optimum temperature for conidial germination and appressorium formation, and characterized penetration and growth of S. turcica in maize leaf sheath and onion epidermis cells, including use of histological staining to assess plant cell viability. The results showed that the optimum temperature for conidial germination and appressorium formation was 20°C. On the maize leaf sheath, the appressoria were formed by germinated conidia, and penetration on the epidermal cells occurred at 8 h postinoculation (hpi). Round vesicles developed beneath the appressoria. Between 16 and 24 hpi, the branched invasive hyphae invaded three to five adjacent cells at most infection sites. The invasive hyphae tended to move along the cell wall and crossed from one cell to another. In the onion epidermis cells, the appressoria formed at 8 hpi, and in most cases the epidermal cells were penetrated through the juncture of the cell walls. At 16–24 hpi, the primary hyphal terminus swelled to a vesicle. The maize leaf sheath cells died at 8 hpi, whereas the onion cells did not. Our findings documented in detail the penetration and invasive hyphal growth in maize leaf sheath and onion epidermis, as well as viability of plant cells, at the early stages of infection, and provide a foundation for elucidating the underlying mechanism of S. turcica–maize interactions.     

Temporal Variation in Setosphaeria turcica Between 1974 and 1994 and Origin of Races 1, 23, and 23N in the United States

ABSTRACT Setosphaeria turcica causes northern leaf blight, an economically important disease of maize throughout the world. Survey collections of S. turcica isolates from 1974 to 1994 provided a unique opportunity to examine temporal diversity in the eastern United States. Two hundred forty-two isolates of S. turcica from maize were studied with random amplified polymorphic DNA (RAPD) markers, mating type, and virulence on maize differential inbred lines with known Ht resistance genes to examine changes over time. One hundred forty-nine RAPD haplotypes were identified. Nearly 20% of haplotypes recurred in more than one year. Race 0 isolates declined in frequency from 83% in 1974 to near 50% in the 1990s, most likely in response to the widespread deployment of Ht1 in commercial maize hybrids. Races 23 and 23N were present in the collection at low levels throughout the study period and were also found among isolates from Virginia in 1957. The frequency of MAT1-2 isolates increased sharply after 1979 and was associated with the emergence of race 1 during the same period. RAPD markers were used to investigate the genetic diversity among a subset of isolates collected in the United States from 1976 to 1982, the period in which this dramatic shift in race frequency occurred. Multilocus haplotypes were not exclusively associated with known races of S. turcica. Based on shared haplotypes and cluster analysis, race 1 isolates share greater similarity with race 0 than with 23 or 23N isolates, indicating race 1 probably evolved from multiple lineages of race 0. Sorghum spp.-infecting isolates share greater similarity with one another than with maize-infecting isolates and represent a distinct subgroup.     

Assessment of early blight (<Emphasis Type="Italic">Alternaria solani</Emphasis>) resistance in tomato using a droplet inoculation method

A droplet inoculation method was used for evaluation of tomato resistance to early blight, a destructive foliar disease of tomato caused by Alternaria solani (Ellis and Martin) Sorauer. In this test method, leaflets are inoculated with small droplets of a spore suspension in either water or a 0.1% agar solution. Early blight resistance was evaluated based on lesion size. The droplet method better discriminated the level of resistance (P < 0.001) for a range of spore densities in comparison with the more commonly used spray inoculation method. Lesions generated by droplet inoculation at 7 days after inoculation ranged from small flecks to almost complete blight with an exponential-like distribution of lesion sizes. Significant correlations (r = 0.52, 0.58, and 0.63, P < 0.001) were observed across three glasshouse tests of 54 accessions including wild species using the droplet method. The most resistant accessions included wild species: one accession of Solanum arcanum, three accessions of Solanum peruvianum, one accession of Solanum neorickii, and one of Solanum chilense. Solanum pennellii and Solanum pimpinellifolium accessions were susceptible, whereas Solanum habrochaites and Solanum lycopersicum accessions ranged from susceptible to moderately resistant. The droplet test method is simple to apply, offers a fine discrimination of early blight resistance levels, and allows objective evaluation.     

Differential interactions ofPhytophthora capsici isolates with pepper genotypes at various plant growth stages

Pepper cultivars from diverse geographic origins were evaluated for resistance to different isolates ofPhytophthora capsici under controlled environmental conditions. All accessions tested were susceptible at the four-leaf stage to the six isolates ofP. capsici. Inoculation at the eight-leaf stage resulted in significantly different interactions among the accessions andP. capsici isolates. The Korean and U.S. cultivars tested were highly susceptible to the isolates ofP. capsici at this stage. In contrast, PI 201234 and PI 201238 had a differential interaction with someP. capsici isolates. At the twelve-leaf stage, Phytophthora blight developed slowly in the Korean and U.S. cultivars that were highly susceptible at the eight-leaf stage. Furthermore, the accessions from the Asian Vegetable Research and Development Center (AVRDC) became highly resistant toP. capsici at this stage.     

山东省玉米生产品种和部分自交系对纹枯病抗性鉴定

连续两年对山东省72份玉米生产品种和30份自交系材料在田间进行纹枯病抗性鉴定,采用被病原菌侵染的高粱粒作为接种物,在玉米拔节中后期接种,按照国家玉米区域试验抗性鉴定标准进行病害分级评定和抗性评价。在102份材料中抗性级别为高抗、抗、中抗、感、高感的比例分别为1.96%、20.59%、24.51%、33.33%、19.61%,生产品种‘登海3号’和‘聊玉22号’对纹枯病表现高抗。     

Levels of fire blight (<Emphasis Type="Italic">Erwinia amylovora</Emphasis>) susceptibility of native apple,pear and quince germplasm from Lake Van Basin,Turkey

Fire blight resistance of apple, pear and quince genetic resources from Lake Van Basin (eastern Turkey) was tested using Erwinia amylovora strain Ea Van. Shoot tips of 92 native accessions (48 accessions for apple, 38 accessions for pear and 6 accessions for quince) were wounded for inoculation, and artificially inoculated with pathogenic bacteria under greenhouse conditions. The levels of resistance of accessions were classified in comparison with control varieties according to the genotype susceptibility index (GSI%) scores based on the lesion length on shoots of each genotype. Fire blight resistance of accessions consisted of five classes: resistant (R), moderately resistant (MR), moderately susceptible (MS), susceptible (S) and highly susceptible (HS). GSI% scores differed significantly among accessions from each fruit species (p < 0.01). GSI values ranged from 12.4% to 64.1% for apple genotypes, from 17.2% to 55.1% for pear genotypes, and from 17.8% to 43.4% for quince genotypes. No resistant genotypes of apple, pear and quince were observed. Seven accessions of apple, two accessions of pear and one accession of quince were MR. 25 accessions of apple, 14 accessions of pear and one accession of quince were MS. These findings indicate a considerable variation in fire blight resistance and could contribute to breeding efforts regarding fire blight resistance in apple, pear and quince.     

Identification and characterization of RB-orthologous genes from the late blight resistant wild potato species Solanum verrucosum

Late blight, caused by the oomycete pathogen, Phytophthora infestans, is a devastating disease of potatoes and tomatoes. A key long-term management strategy for combating this disease is to develop potato cultivars with high levels of durable resistance through identification and integration of major resistance genes. The RBgene, cloned from the Mexican diploid potato species Solanum bulbocastanum, confers broad-spectrum resistance to potato late blight. Here, we have determined the late blight resistance phenotypes of eight accessions of Solanum verrucosum, another wild diploid potato species, using greenhouse inoculations and discovered variability among the accessions. While most accessions were resistant, one accession was notably more susceptible. Transcribed orthologs of the RB gene from the eight S. verrucosum accessions were cloned using a homology-based PCR approach. Sequence analysis revealed that the RB^ver orthologs share up to 83.5% nucleotide identity with RB^blb. Stable introduction of the RB ortholog from late blight resistant S. verrucosum PI275260 into susceptible S. tuberosum confers resistance to P. infestans. Interestingly, this functional RB^ver ortholog contains an insertion of a complete leucine rich repeat when compared to RB^blb, and differs from the RB^ver ortholog from a susceptible accession at only four amino acid residues.     

玉米大斑病菌人工接种方法研究

 The efficient and accurate method of inoculating Exserohilum turcicum on maize is most important for studying the corn leaf blight. In this study, four approaches including spore suspension spraying, inoculation with grinding infected maize leaves, inoculation liquid inoculum into the leaf whorl and inoculation with inoculated sorghum seeds were compared with natural induction in fields during different growth stages of maize in the greenhouse and field. The results showed that disease incidence of all varieties tested were up to 100% by inoculating with inoculated sorghum seeds during the booting stage (11 to 12 leaf stages). This method will not only guarantee the enough pathogen inocula but also be easy to use, and could be an effective way for disease resistance identification in field and greenhouse.      

A detailed evaluation method to identify sources of quantitative resistance to Fusarium oxysporum f. sp. pisi race 2 within a Pisum spp. germplasm collection

Fusarium oxysporum f. sp. pisi (Fop) is an important pathogen of field pea (Pisum sativum) worldwide. The constant evolution of the pathogen drives the necessity to broaden the genetic basis of resistance to Fop. To achieve this, it is important to have a large germplasm collection available and an accurate and efficient method for disease assessment. Here, a detailed evaluation method coupling disease incidence, disease rating over time and its related area under the disease progression curve (AUDPC) was established and used to screen a Pisum spp. germplasm collection against one isolate of Fop race 2. A large variation in the disease response of specific pea accessions ranging from highly resistant to susceptible was observed within the collection, indicating the quantitative expression of the resistance. The repetition of the inoculation experiments on a subset of 19 accessions, including two susceptible accessions, indicated that the scoring method was robust and reproducible and confirmed the highly resistant phenotypes of 11 accessions. To initiate the characterization of resistance mechanisms within these accessions, the external and internal stem symptoms were compared between these selected pea accessions, together with the extent of fungal colonization within plants. All these tests indicated that, in all resistant accessions, the resistance mechanisms efficiently stopped pathogen progression at the crown. Incorporation of these sources of resistance to breeding programmes will contribute to improved Fop resistance in pea cultivars.     

Sources of resistance to ascochyta blight in wild Cicer species

Ascochyta blight [Ascochyta rabiei (Pass.) Lab.] is the major foliar disease of chickpea (Cicer arietinum L.). In search of better sources of resistance to ascochyta blight, 201 accessions of 8 annual wildCicer species were evaluated in field and greenhouse for 3 years (1988 to 1991) at Tel Hadya, Syria. One accession each ofC. judaicum Boiss (ILWC 165) andC. pinnatifidum Jaub. & Spach. (ILWC 159) were consistently rated resistant in both field and greenhouse evaluations. Another three accessions ofC. judaicum (ILWC 61, ILWC 154, ILWC 199) and six accessions ofC. pinnatifidum (ILWC 78, ILWC 88, ILWC 155, ILWC 160, ILWC 162, ILWC 203) were resistant or moderately resistant. The blight-resistant accessions ofC. judaicum originated from Jordan, Lebanon, Syria, and Turkey; and those ofC. pinnatifidum from Syria and Turkey. None of the accessions ofC. bijugum, C. chorassanicum, C. cuneatum, C. echinospermum, C. reticulatum andC. yamashitae were resistant to blight.     

Genetic diversity of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan based on phylogenetic analyses of rDNA-IGS and <Emphasis Type="Italic">MAT1</Emphasis> sequences

Twenty-eight isolates of Fusarium oxysporum f. sp. spinaciae (FOS; the causal agent of spinach wilt) collected from Japan were assessed for mating type and subjected to phylogenetic analysis. Mating type analysis revealed all isolates to be MAT1-2, suggesting that there is no sexual recombination within the population. Phylogenetic analyses based on nucleotide sequences of the ribosomal DNA intergenic spacer (IGS) and the mating type locus (MAT1) suggested that FOS is polyphyletic. The cluster analysis based on IGS showed four phylogenetic groups (S1–S4) among the isolates. Two distinct lineages, S1 and S3, included FOS isolates both of the vegetative compatibility group (VCG) types, 0330 and 0331, demonstrating that VCG differentiation in FOS may not necessarily reflect the phylogenetic relationships based on IGS and MAT1-2-1.     

Identification and pathogenicity of Fusarium species associated with pokkah boeng of sugarcane in Brazil

Brazil is the world’s biggest producer of sugarcane (Saccharum spp. hybrids). Pokkah boeng is an important fungal disease in this crop caused by members of the Fusarium fujikuroi species complex (FFSC) and characterized by deformation of the aerial part of the plant and stem rot. While the occurrence of symptoms has been reported in plantations in Brazil, no official reports of the disease exist. In this study, species of the FFSC were identified that are associated with sugarcane plants with symptoms of pokkah boeng in Brazil. This was achieved using two-loci molecular phylogeny, sexual compatibility and analysis of morphological markers. The ability of strains to cause disease in plants of sugarcane, maize, sorghum and millet was also evaluated. The 39 isolates studied were identified as F. sacchari, F. proliferatum and another, still unknown, phylogenetic lineage that is sister to F. andiyazi. Crossing field isolates of F. sacchari and F. proliferatum with their respective tester strains produced fertile perithecia and viable ascospores. All three species induced symptoms of pokkah boeng on inoculated sugarcane plants and caused stem rot in maize, sorghum and millet. Symptoms on sugarcane are chlorosis and necrosis of leaves, punctured lesions, twisted leaves, reduction of the total leaf area, death of the top of the plant and stalk rot. The findings confirmed the aetiology of the disease in Brazil, generated basic knowledge for the development of strategies for diagnosis and monitoring of the disease and support breeding programmes for selecting resistant germplasm.     

Anthracnose of <Emphasis Type="Italic">Polygonatum falcatum</Emphasis> caused by <Emphasis Type="Italic">Colletotrichum dematium</Emphasis>

Severe spotting, blight and drop of leaves caused by Colletotrichum dematium were found on potted plants of Polygonatum falcatum, a liliaceous ornamental, in open fields in Kagawa Prefecture, Japan, in May 2001. This new disease was named anthracnose of P. falcatum. Keisuke Tomioka, Jouji Moriwaki, Toyozo Sato contributed equally to this work. The fungal isolate and its nucleotide sequence data obtained in this study were deposited in Genebank, National Institute of Agrobiological Sciences and the DDBJ/EMBL/GenBank databases under accessions MAFF239500 and AB334523, respectively.     

凸脐蠕孢菌玉米、高粱专化型的细胞壁降解酶活性及基因表达分析

为探明玉米专化型和高粱专化型凸脐蠕孢菌的细胞壁降解酶在致病过程中的作用,采用酶活性检测方法测定了2种专化型的细胞壁降解酶活性,并检测了相关基因的表达。结果表明:高粱专化型凸脐蠕孢菌的聚甲基半乳糖醛酸酶(PMG)活性为115.84 U/mg,略高于玉米专化型;玉米专化型的多聚半乳糖醛酸酶(PG)和纤维素酶(Cx)的活性分别为151.76 U/mg和168.53 U/mg,略高于高粱专化型;且同一种专化型菌株的细胞壁降解酶活性存在差异。2种专化型的细胞壁降解酶基因表达量存在差异,Cx基因在2种专化型互作过程中均随病程的延长而大幅度上调表达;高粱专化型的PG基因随病程的延长大幅度上调表达,而玉米专化型的PG基因随病程的延长上调表达量有所下降;高粱专化型的PMG基因随病程的延长大幅度上调表达,而玉米专化型的PMG基因随病程的延长下调表达。推测产酶能力、基因表达和基因时间表达的差异可能是引起凸脐蠕孢菌专化型致病专化性的诱因之一。     

Aggressiveness of eight <Emphasis Type="Italic">Didymella rabiei</Emphasis> isolates from domesticated and wild chickpea native to Turkey and Israel,a case study

Ascochyta blight, caused by Didymella rabiei, affects both domesticated chickpea and its congeneric wild relatives. The aim of this study was to compare the aggressiveness of D. rabiei isolates from wild and domesticated Cicer spp. in Turkey and Israel on wild and domesticated hosts from both countries. A total of eight isolates of D. rabiei sampled from C. pinnatifidum, C. judaicum and C. arietinum in Turkey and Israel was tested on two domesticated chickpea cultivars and two wild Cicer accessions from Turkey and Israel. Using cross-inoculation experiments, we compared pathogen aggressiveness across the different pathogen and host origin combinations. Two measures of aggressiveness were used, incubation period and relative area under the disease progress curve. The eight tested isolates infected all of the host plants, but were more aggressive on their original hosts with one exception; Turkish domesticated isolates were less aggressive on their domesticated host in comparison to the aggressiveness of Israeli domesticated isolates on Turkish domesticated chickpea. C. judaicum plants were highly resistant against all of the isolates from different origins except for their own isolates. Regardless of the country of origin, the wild isolates were highly aggressive on domesticated chickpea while the domesticated isolates were less aggressive on the wild hosts compared with the wild isolates. These results suggest that the aggressiveness pattern of D. rabiei on different hosts could have been shaped by adaptation to the distinct ecological niches of wild vs. domesticated chickpea.     

Differences in the responses of melon accessions to fusarium root and stem rot and their colonization by Fusarium oxysporum f. sp. radicis‐cucumerinum

Fusarium root and stem rot caused by the fungus Fusarium oxysporum f. sp. radicis‐cucumerinum is a major disease in greenhouse cucumbers. Over the past decade, the disease has been documented in melon greenhouses in Greece, and recently it has been sporadically recorded in greenhouse melons in Israel. Variations in disease response were found among 41 melon accessions artificially inoculated with the pathogen: 10 accessions were highly susceptible (90–100% mortality), 23 exhibited an intermediate response (20–86%) and eight were resistant (0–4%). Two melon accessions – HEM (highly resistant) and TAD (partially resistant) – were crossed with the susceptible accession DUL. The responses of the three accessions and F₁ crosses between the resistant and susceptible parents were evaluated. HEM contributed higher resistance to the F₁ hybrid than TAD. Roots of susceptible and resistant accessions were 100 and 79% colonized, respectively, following artificial inoculation. However, only susceptible plants showed colonization of the upper plant tissues. Microscopic evaluation of cross sections taken from the crown region of the susceptible DUL revealed profuse fungal growth in the intercellular spaces of the parenchyma and in xylem vessels. In the resistant cultivar HEM, very little fungal growth was detected in the intercellular spaces of the parenchyma, and none in the xylem or any other vascular tissue. Finding resistant accessions may create an opportunity to study the genetics of resistance inheritance and to develop molecular markers that will facilitate breeding resistant melon cultivars.