Bacterial Streak Disease of Maize (Zea mays L.) in South Africa

Bacterial streak disease of maize is currently causing some concern among breeders in South Africa. The causal organism of this previously undescribed disease was successfully isolated and its pathogenicity established using KoCH's postulates. Standard physiological and biochemical tests used to identify phytopathogenic bacteria indicated that the bacterium is a Xanthomonas campestris pathovar. Comparisons between this organism and other recognized X. campestris pathovars of the Poaceae indicated that apart from some minor differences the maize streak pathogen is physiologically similar to X. campestris pv. holcicola. However, in repeated reciprocal inoculation experiments all attempts to induce disease symptoms in sorghum with the maize streak pathogen were unsuccessful. Conversely, X. campestris pv. holcicola did produce symptoms in maize leaves. In all the maize cultivars tested the symptoms produced by the maize streak pathogen were, however, always considerably more severe than those caused by X. campestris pv. holcicola. Notwithstanding its physiological similarity to X. campestris pv. holicola it would appear that on the grounds of host specificity the maize streak pathogen warrants new pathovar status. The name X. campestris pv. zeae is proposed.     

Survival and retention of infectivity of bacterial stalk rot pathogen of maize and its perpetuation on varied cropping pattern

Summary The sterilized environment prompted the maize stalk rot pathogen Erwinia carotovora f. sp. zeae Sabet to retain its infectivity for a longer period than the unsterilized one. The survival as well as infectivity potency of the pathogen present in the soil having naturally-infested or artificially-inoculated host plant as debris was superior to that in the soil or even the soil containing maize stalk tissue. ‘Maize-potato-maize’ cropping pattern effected a considerable increase in the disease incidence due to maintainance of continuity of life of the pathogen in the intervening potato crop. re]19741113     

Antifungal metabolites (monorden, monocillins I, II, III) from Colletotrichum graminicola, a systemic vascular pathogen of maize

Colletotrichum graminicola is a systemic vascular pathogen that causes anthracnose stalk rot and leaf blight of maize. In the course of an effort to explore the potential presence and roles of C. graminicola metabolites in maize, ethyl acetate extracts of solid substrate fermentations of several C. graminicola isolates from Michigan and Illinois were found to be active against Aspergillus flavus and Fusarium verticillioides, both mycotoxin-producing seed-infecting fungal pathogens. Chemical investigations of the extract of one such isolate (NRRL 47511) led to the isolation of known metabolites monorden (also known as radicicol) and monocillins I–III as major components. Monorden and monocillin I displayed in vitro activity against the stalk- and ear-rot pathogen Stenocarpella maydis while only the most abundant metabolite (monorden) showed activity against foliar pathogens Alternaria alternata, Bipolaris zeicola, and Curvularia lunata. Using LC–HRESITOFMS, monorden was detected in steam-sterilized maize stalks and stalk residues inoculated with C. graminicola but not in the necrotic stalk tissues of wound-inoculated plants grown in an environmental chamber. Monorden and monocillin I can bind and inhibit plant Hsp90, a chaperone of R-proteins. It is hypothesized that monorden and monocillins could support the C. graminicola disease cycle by disrupting maize plant defenses and by excluding other fungi from necrotic tissues and crop residues. This is the first report of natural products from C. graminicola, as well as the production of monorden and monocillins by a pathogen of cereals.     

Detection and Molecular Diversity of Pantoea ananatis Associated with White Spot Disease in Maize,Sorghum and Crabgrass in Brazil

Bacteria of the genus Pantoea have become important plant pathogens worldwide in recent years. Pantoea ananatis was reported as the cause of maize white spot, a serious maize disease in Brazil, causing significant yield losses. However, very little information is available about how to detect this pathogen, its genetic variability and the putative alternative hosts in maize‐growing areas. To address these issues, we implemented a rapid and efficient PCR‐based method to identify P. ananatis isolated from leaves showing white spot symptoms and evaluated its genetic diversity in maize, sorghum and crabgrass. Of the 29 bacteria isolated from typical water‐soaked lesions of white spot disease that produced yellow colonies, 15 isolates were identified as P. ananatis by 16S rDNA sequencing and correctly detected by the PCR reaction, amplifying a specific fragment of the ice nucleation gene (ina). These P. ananatis isolates included 13 from maize, one from sorghum and one from crabgrass, while the other 14 yellow colony isolates were from other bacterial species, including two Pantoea species (Pantoea dispersa and Pantoea agglomerans) that were not amplified by the ina primers. These results indicate that the optimized PCR assay can be used to detect P. ananatis isolated from white spot lesions and could be used as a large‐scale and cost‐effective method of detecting this pathogen in leaf lesions on maize and other grasses. All isolates were evaluated for hypersensitive response (HR) on tobacco, revealing that some P. ananatis were able to induce HR. The high genetic variability revealed by rep‐PCR did not differentiated the P. ananatis isolates based on their hosts or HR reaction. The detection, characterization and diversity of P. ananatis from maize, sorghum and crabgrass in our study can be applied in understanding epidemiology and designing control strategies for maize white spot disease in Brazil.     

Rhizospheric Enterobacter enhanced maize seedling health and growth

Phyto-beneficial effects of rhizobacteria specifically Enterobacter species were evaluated on maize seedling health and growth. Out of the 19 isolates examined, two were remarkable in their phosphate solubilization efficiency (PSE > 70%), chitinase enzyme activity (CEA > 85%) and antifungal activity with evidence of no or low disease expression in maize seedling. The selected isolates (OSR7 and IGGR11) were identified and 16S rDNA revealed both isolates as Enterobacter species. Re-evaluation of both isolates ascertains that their combined effects are more effective on maize seedling than their individual effects. Their combinations completely suppressed pathogenic activity of Fusarium verticillioides on maize seedling with evidence of no disease symptoms. Other treatments significantly (p < 0.05) expressed varied maize seedling diseases such as leaf curl and stem rot. Apart from treatment T2 (maize + pathogen), other treatments most especially their combinations significantly (p < 0.05) enhanced seedling height, stem girth, leaf area, nitrogen and potassium contents. The phyto-beneficial effects of these Enterobacter species suggest that they could be employed as bio-inoculant for maize seedling health and growth.     

Clavibacter nebraskensis causing Goss's wilt of maize: Five decades of detaining the enemy in the New World

Goss's bacterial wilt and leaf blight of maize (Zea mays) caused by the gram-positive coryneform bacterium Clavibacter nebraskensis is an economically important disease in North America. C. nebraskensis is included within the high-risk list of quarantine pathogens by several plant protection organizations (EPPO code: CORBMI), hence it is under strict quarantine control around the world. The causal agent was reported for the first time on maize in Nebraska (USA) in 1969. After an outbreak during the 1970s, prevalence of the disease decreased in the 1980s to the early 2000s, before the disease resurged causing a serious threat to maize production in North America. The re-emergence of Goss's wilt in the corn belt of the United States led to several novel achievements in understanding the pathogen biology and disease control. In this review, we provide an updated overview of the pathogen taxonomy, biology, and epidemiology as well as management strategies of Goss's wilt disease. First, a taxonomic history of the pathogen is provided followed by symptomology and host range, genetic diversity, and pathogenicity mechanisms of the bacterium. Then, utility of high-throughput molecular approaches in the precise detection and identification of the pathogen and the management strategies of the disease are explained. Finally, we highlight the role of integrated pest management strategies to combat the risk of Goss's wilt in the 21st century maize industry.

Disease symptoms

Large (2–15 cm) tan to grey elongated oval lesions with wavy, irregular water-soaked margins on the leaves. The lesions often start at the leaf tip or are associated with wounding caused by hail or wind damage. Small (1 mm in diameter), dark, discontinuous water-soaked spots, known as “freckles”, can be observed in the periphery of lesions. When backlit, the freckles appear translucent. Early infection (prior to growth stage V6) may become systemic and cause seedlings to wilt, wither, and die. Coalescence of lesions results in leaf blighting.

Host range

Maize (Zea mays) is the only economic host of the pathogen. A number of Poaceae species are reported to act as secondary hosts for C. nebraskensis.

Taxonomic status of the pathogen

Class: Actinobacteria; Order: Micrococcales; Family: Microbacteriaceae; Genus: Clavibacter; Species: Clavibacter nebraskensis.

Synonyms

Corynebacterium nebraskense (Schuster, 1970) Vidaver & Mandel 1974; Corynebacterium michiganense pv. nebraskense (Vidaver & Mandel 1974) Dye & Kemp 1977; Corynebacterium michiganense subsp. nebraskense (Vidaver & Mandel 1974) Carlson & Vidaver 1982; Clavibacter michiganense subsp. nebraskense (Vidaver & Mandel 1974) Davis et al. 1984; Clavibacter michiganensis subsp. nebraskensis (Vidaver & Mandel 1974) Davis et al. 1984.

Type materials

ATCC 27794^T; CFBP 2405^T; ICMP 3298^T; LMG 3700^T; NCPPB 2581^T.

Microbiological properties

Cells are gram-positive, orange-pigmented, pleomorphic club- or rod-shaped, nonspore-forming, nonmotile, and without flagella, approximately 0.5 × 1–2.0 μm.

Distribution

The pathogen is restricted to Canada and the United States.

Phytosanitary categorization

EPPO code CORBNE.     

Distribution,Etiology, Molecular Genetics and Management Perspectives of Northern Corn Leaf Blight of Maize (<i>Zea mays</i> L.)

Maize is cultivated extensively throughout the world and has the highest production among cereals. However, Northern corn leaf blight (NCLB) disease caused by Exherohilum turcicum, is the most devastating limiting factor of maize production. The disease causes immense losses to corn yield if it develops prior or during the tasseling and silking stages of crop development. It has a worldwide distribution and its development is favoured by cool to moderate temperatures with high relative humidity. The prevalence of the disease has increased in recent years and new races of the pathogen have been reported worldwide. The fungus E. turcicum is highly variable in nature. Though different management strategies have proved effective to reduce economic losses from NCLB, the development of varieties with resistance to E. turcicum is the most efficient and inexpensive way for disease management. Qualitative resistance for NCLB governed by Ht genes is a race-specific resistance which leads to a higher level of resistance. However, some Ht genes can easily become ineffective under the high pressure of virulent strains of the pathogen. Hence, it is imperative to understand and examine the consistency of the genomic locations of quantitative trait loci for resistance to NCLB in diverse maize populations. The breeding approaches for pyramiding resistant genes against E. turcicum in maize can impart NCLB resistance under high disease pressure environments. Furthermore, the genome editing approaches like CRISPR-cas9 and RNAi can also prove vital for developing NCLB resistant maize cultivars. As such this review delivers emphasis on the importance and current status of the disease, racial spectrum of the pathogen, genetic nature and breeding approaches for resistance and management strategies of the disease in a sustainable manner.     

Cotton,cowpea and sesame are alternative crops to cucurbits in soils naturally infested with Monosporascus cannonballus

Monosporascus cannonballus is an important cucurbit root pathogen, which has been reported in the main production areas of melon and watermelon in Brazil and worldwide and potentially capable to colonize roots of different species. Crop rotation is considered an effective management strategy to prevent this disease. The aim of this study was to evaluate the response of different crops, pumpkin, cotton, cowpea, sesame, watermelon, melon, corn, cucumber, sorghum and tomato, to the infection of this pathogen. Seedlings were transplanted into plastic containers with an inoculum concentration of 20 colony‐forming units (CFU) g^?1 of M. cannonballus. Fifty days after transplanting, the variables analysed were the degree of disease severity on the root system and the frequency of reisolation. On cucurbits, the results demonstrated different degrees of susceptibility among crops and cultivars, being melon and watermelon the most sensitive species. In contrast, Cucurbita cultivars were the most tolerant. Regarding non‐cucurbit crops, maize, sorghum and tomato presented root discoloration and M. cannonballus was reisolated from roots. Cotton, cowpea and sesame cultivars were not affected by the pathogen, so they can be considered as alternative crops to be cultivated, or in rotation with cucurbits, in M. cannonballus infested soils.     

I. Pathogenic variation in fungi and bacteria and mycorrhizal compatibility: The genetic architecture of aggressiveness in Ustilago maydis

Isolates of Ustilago maydis (D.C.) Cda are known to differ in the degree to which they can parasitise maize and, as no differential reaction occurs between pathogen and host, these differences must be differences in aggressiveness. Inoculation of juvenile maize plants induced morphological changes which provided a means of assessing and distinguishing U. maydis isolates in respect of their aggressiveness. Thus it was possible to carry out a survey of the genetic architecture of aggressiveness in natural populations of the pathogen. The distribution of aggressiveness within a single gall suggested that selection had taken place over a number of generations in favour of genes or gene systems tending to increase aggressiveness. Diallele analyses of isolates from two natural populations revealed that aggressiveness had a low heritability and was determined by genes exhibiting mainly non-allelic interaction. It has been proposed that horizontal resistance in host cultivars would have a more long lasting protective effect against disease. However, the genetic architecture of aggressiveness in U. maydis should enable the pathogen to respond rapidly to changes in host resistance and the skewed distribution found within a single gall tends to support this. Therefore, the use of higher levels of host resistance should rapidly select for higher aggressiveness in the pathogen.     

Use of virus-induced gene silencing to characterize genes involved in modulating hypersensitive cell death in maize

Plant disease resistance proteins (R-proteins) detect specific pathogen-derived molecules, triggering a defence response often including a rapid localized cell death at the point of pathogen penetration called the hypersensitive response (HR). The maize Rp1-D21 gene encodes a protein that triggers a spontaneous HR causing spots on leaves in the absence of any pathogen. Previously, we used fine mapping and functional analysis in a Nicotiana benthamiana transient expression system to identify and characterize a number of genes associated with variation in Rp1-D21-induced HR. Here we describe a system for characterizing genes mediating HR, using virus-induced gene silencing (VIGS) in a maize line carrying Rp1-D21. We assess the roles of 12 candidate genes. Three of these genes, SGT1, RAR1, and HSP90, are required for HR induced by a number of R-proteins across several plant–pathogen systems. We confirmed that maize HSP90 was required for full Rp1-D21-induced HR. However, suppression of SGT1 expression unexpectedly increased the severity of Rp1-D21-induced HR while suppression of RAR1 expression had no measurable effect. We confirmed the effects on HR of two genes we had previously validated in the N. benthamiana system, hydroxycinnamoyltransferase and caffeoyl CoA O-methyltransferase. We further showed the suppression the expression of two previously uncharacterized, candidate genes, IQ calmodulin binding protein (IQM3) and vacuolar protein sorting protein 37, suppressed Rp1-D21-induced HR. This approach is an efficient way to characterize the roles of genes modulating the hypersensitive defence response and other dominant lesion phenotypes in maize.     

Expression of a maize proteinase inhibitor gene is induced in response to wounding and fungal infection: systemic wound-response of a monocot gene

The isolation and characterization of cDNA and genomic clones encoding a proteinase inhibitor protein (MPI) in maize is reported. Accumulation of the MPI mRNA is induced in response to fungal infection in germinating maize embryos. The expression pattern of the MPI gene, in healthy and fungal infected maize tissues, was examined and compared with the expression pattern of a gene that codes for a pathogenesis-related protein (the PRms protein) from maize. These two genes are induced by fungal infection, however different signals trigger their activation. Accumulation of the proteinase inhibitor mRNA is more a consequence of the wound produced by the penetration and colonization of the host tissues by the pathogen, than the result of a direct molecular recognition of the pathogen by the plant, as is the case for the induction of the PRms gene. Wounding, or treatment with abscisic acid or methyl jasmonate, stimulate MPI mRNA accumulation, but not PRms mRNA accumulation. Local and systemic induction of the MPI gene expression in response to wounding occurs in maize plants. To the authors' knowledge, this is the first example of a gene from a monocotyledonous species that clearly shows a systemic wound response. The possible functional implications for the existence of different signal transduction pathways that simultaneously activate a battery of defense mechanisms against potential pathogens are discussed.     

Understanding resistant germplasm‐induced virulence variation through analysis of proteomics and suppression subtractive hybridization in a maize pathogen Curvularia lunata

Curvularia lunata is an important pathogen causing Curvularia leaf spot in maize. Significant pathogenic variation has been found in C. lunata. To better understand the mechanism of this phenomenon, we consecutively put the selective pressures of resistant maize population on C. lunata strain WS18 (low virulence) artificially. As a result, the virulence of this strain was significantly enhanced. Using 2DE, 12 up‐regulated and four down‐regulated proteins were identified in virulence‐increased strain compared to WS18. Our analysis revealed that melanin synthesis‐related proteins (Brn1, Brn2, and scytalone dehydratase) and stress tolerance‐related proteins (HSP 70) directly involved in the potential virulence growth as crucial markers or factors in C. lunata. To validate 2DE results and screen differential genes at mRNA level, we constructed a subtracted cDNA library (tester: virulence‐increased strain; driver: WS18). A total of 188 unigenes were obtained this way, of which 14 were indicators for the evolution of pathogen virulence. Brn1 and hsp genes exhibited similar expression patterns corresponding to proteins detected by 2DE. Overall, our results indicated that differential proteins or genes, being involved with melanin synthesis or tolerance response to stress, could be considered as hallmarks of virulence increase in C. lunata.     

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

Various chitinases have been shown to inhibit the growth of fungal pathogens in in vitro as well as in planta conditions. chi194, a wheat chitinases gene encoding a 33-kDa chitinase protein, was overexpressed in tomato plants (cv. Pusa Ruby) under the control of maize ubiquitin 1 promoter. The integration of transgene in tomato plants was confirmed with polymerase chain reaction (PCR) and Southern blot analysis. The inheritance of the transgene in T₁ and T₂ generations were shown by molecular analysis and the hygromycin sensitivity test. The broad range of chitinase activity was observed among the transgenic lines in T₀ and a similar range was retained in the T₁ and T₂ generations. Most importantly, the transgenic tomato lines with high chitinase activity were found to be highly resistant to the fungal pathogen Fusarium oxysporum f. sp. lycopersici. Thus, the results demonstrated that the expression of the wheat endochitinase chi194 in tomato plants confers resistance against Fusarium wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. lycopersici.     

Biological Control of Stenocarpella maydis in Maize Seed with Antagonistic Streptomyces sp. Isolates

The effectiveness of two Streptomyces sp. isolates, isolated from maize rhizosphere soil and designated as DAUFPE 11470 and DAUFPE 14632, was evaluated in vitro and under greenhouse conditions for control of Stenocarpella maydis in maize seeds. Stenocarpella maydis incidence was detected in all subsamples of disease‐free maize seeds by in vitro survey test, and ranged from 10.8% to 65.2%. In a filter paper test with surface‐disinfected seeds inoculated with S. maydis, Streptomyces sp. isolates DAUFPE 11470 and DAUFPE 14632 significantly reduced (P ≤ 0.05) the pathogen incidence by 93.2% and 92.3%, respectively. Seed germination in the same treatments was increased by 30.0% and 28.2%, respectively. Treatments of non‐disinfected seeds with the isolates DAUFPE 11470 and DAUFPE 14632, under greenhouse conditions reduced disease incidence in the seedlings by 87.3% and 85.6%, respectively. The reductions in disease incidence in surface‐disinfected seeds were 85.0% and 83.0% for the same isolates. Seedling emergence significantly (P ≤ 0.05) increased in disinfected and non‐disinfected seeds inoculated with the Streptomyces sp. isolates. The results indicate the potential of using Streptomyces sp. isolates as an additional tool to control Stenocarpella ear rot by significantly reducing the incidence of S. maydis in maize seeds and seedlings.     

Survival in Dalbulus leafhopper vectors improves after exposure to maize stunting pathogens

Using D. maidis and pathogen isolates collected at three field sites along an altitudinal gradient in Mexico, we compared survival in leafhoppers exposed to healthy maize to those exposed to maize infected with one of four isolates of maize stunting pathogens: two isolates of the corn stunt spiroplasma (CSS, Spiroplasma kunkelii) and two of the maize busby stunt phytoplasma (MBSP). Survival improved after exposure to either plant pathogen under both the cooler and warmer environmental conditions D. maidis is likely to encounter during the dry season. Survival varied among leafhoppers from the different field sites, suggesting that gene flow between these populations is limited. The leafhoppers responded differently to the four isolates (i.e., we noted significant population by exposure interactions), but we found no difference between MBSP and CSS exposure. Finally, we found evidence of local adaptation in one leafhopper population to sympatric, as compared to allopatric, plant pathogens. We have shown with this and our earlier study that aspects of the interaction phenotype in the association between D. maidis and the plant pathogens are mutualistic and that this association has considerable potential as a model for studies of local adaptation.     

Ustilago maydis Produces Cytokinins and Abscisic Acid for Potential Regulation of Tumor Formation in Maize

The infection of maize (Zea mays) by the basidiomycete fungus Ustilago maydis leads to common smut of corn characterized by the production of tumors in susceptible aboveground plant tissues. LC-(ES)MS/MS profiles of abscisic acid (ABA) and 12 different cytokinins (CKs) were determined for infected and uninfected maize tissues over a time course following fungal exposure. Samples were taken at points corresponding to the appearance of disease symptoms. Axenic cultures of haploid and dikaryon forms of U. maydis were also profiled. This study confirmed the capability of Ustilago maydis to synthesize CKs, ABA, and auxin (IAA). It also provided evidence for the involvement of CK and ABA in the U. maydis-maize infection process. Significant quantities of CKs and ABA were detected from axenic cultures of U. maydis as was IAA. CKs and ABA levels were elevated in leaves and stems of maize after infection; notable was the high level of cis-zeatin 9-riboside. Variation among hormone profiles of maize tissues was observed at different time points during infection and between infections with nonpathogenic haploid and pathogenic dikaryon strains. This suggested that CKs and ABA accumulate and are likely metabolized in maize tissue infected with U. maydis. Because U. maydis produced these phytohormones at significant levels, it is possible that the fungal pathogen is a source of these compounds in infected tissue. This is the first study to confirm the production of CKs and document the production of ABA by U. maydis. This study also established an involvement of these phytohormones and a possible functional role for ABA in U. maydis infection of maize.     

Sequence variation of intergenic mitochondrial DNA spacers (mtDNA‐IGS) of Phytophthora infestans (Oomycetes) and related species

The potato late‐blight disease is caused by the pseudofungus Phytophthora infestans (Oomycetes). This pathogen was of historical importance as it caused the Irish Potato Famine. There is currently a worldwide resurgence of the disease. Following worldwide migrations as well as being able to discriminate P. infestans from related species are key issues. We present sequence variation of five inter‐genic mitochondrial DNA spacers (mtDNA‐IGS) for P. infestans and four related taxa. Intra and inter‐taxon variation was observed showing potential for both molecular ecology and molecular systematic.