Identification of a biocontrol agent Bacillus vallismortis BV23 and assessment of effects of its metabolites on Fusarium graminearum causing corn stalk rot

Corn stalk rot, caused by Fusarium graminearum, is one of the most destructive diseases of maize in many regions of the world. A bacterial strain BV23 was isolated from corn rhizosphere that reduced corn stalk rot significantly in greenhouse studies in 2016 and 2017. BV23 was identified as Bacillus vallismortis, which showed antagonistic effects against a number of fungal pathogens, including F. graminearum, Rhizoctonia solani, Athelia rolfsii, and Thanatephorus cucumeris. BV23 had the greatest fungistatic effect on F. graminearum, inhibiting mycelial growth by 66.2%, conidial germination by 90.1%, and conidial production by 86.7%. The probable antifungal mechanism was assessed by examining the morphology and ultrastructure of F. graminearum hyphae. Treatment by BV23 culture supernatant resulted in coarser hyphae, induced cytoplasmic granulation, and increased cell membrane permeability of F. graminearum, causing cytoplasm leakage. These effects became increasingly obvious with increasing concentration (1%, 5% and 10%). Furthermore, the antifungal active substances were sensitive to heat.     

Stalk Rot of Maize: Host-pathogen Interaction

Four maize cultivars were each separately inoculated with 10 different isolates of Diplodia zeae, Fusarium moniliforme, F. graminearum and Macrophomina phaseolina. D. zeae consistently caused most stalk rot, followed in decreasing order by M. phaseolina, F. graminearum and F. moniliforme. Significant intraspecific differences in stalk rot were recorded. No significant stalk rot x cultivar interaction occurred. Stalk rot correlated non-significantly with yield loss but significantly with cellulase production in vitro.     

Effect of soil solarization on corn stalk rot

Seven weeks solarization of irrigated soil raised its temperature by 11.5°C over non-solarized soil at 10 cm depth and effectively controlled weeds (98.5%), stalk borer (8.9%) and stalk rot disease (69.1%) in corn. Solarization also reduced symptoms of Fusarium moniliforme and Macrophomina phaseolina significantly by 64.2% and 78.4%, respectively, and completely controlled M. phaseolina in corn cultivars, viz. Pool-10, Shaheen and Gauher. Whereas symptoms of F. moniliforme were observed in these cultivars, Fusarium graminearum was not observed except in two cultivars, Shaheen and Akbar. Growth of crop planted in solarized plots was better and it yielded almost one to three times more grains in cultivars under test. Soil analysis immediately following solarization revealed that essential elements were readily available in simpler forms, which may have increased pest resistance and reduced stalk breakage.     

Pathogenicity of Fusarium graminearum Chemotypes Towards Corn, Wheat, Triticale and Rye

The pathogenicity of nine Fusarium graminearum isolates belonging to three different trichothecene-producing chemotypes, i.e. NIV + FUS, DON + 3-AcDON, and DON + 15-AcDON was tested on corn, wheat, triticale and rye. Although these compounds are known to have different toxicity, no difference in pathogenicity was observed among the mean values for chemotypes. However, within each chemotype there were isolates with different pathogenicity which was not correlated with the ability to produce trichothecenes in vitro. The pathogenicity of each isolate was almost the same for all tested cereals. The involvement of trichothecenes in pathogenesis was investigated in infected corn stalk tissues. All the expected trichothecenes were detected with the exception of NIV and FUS.     

Use of molecular markers for monitoring fungi involved in stalk rot of corn

Summary Separate genes conferring antibiotic drag resistance have been inserted into Fusarium graminearum and Fusarium moniliforme. These organisms are associated with stalk rot of corn, a disease of uncertain cause. Antibiotic resistant fungi were obtained by developing a gene transfer system using whole cells as recipients for DNA. Hygromycin B and benomyl-resistant colonies were isolated by treating fungal tissue with lithium acetate and adding plasmid vectors containing the respective genes which give drug resistance. The DNA was stably integrated into the fungal chromosome. Following plant inoculation, disease symptoms developed and the isolates were recovered on selective medium. In each case, these fungi retained the transformed phenotype, although extensive rearrangements and/or deletions occurred. Specific molecular tagging allows detailed studies of this interaction and should be of general use in situations involving complex multiple pathogen diseases.     

Potential bio-control agent for management of post-flowering stalk rot complex and maize plant health

The of aim of this study was to evaluate the potentiality of the native isolate of Trichoderma harzianum from IARI farm for the management of post-flowering stalk rot of maize and improvement of plant health. The treatment was tried as a seed treatment with the potent commercial formulation Kalisena SD (Aspergillus niger), neem kernel powder and effective fungicides (Thiram, Carbendazim, Captan) for comparison of its efficacy. The minimum disease was recorded in plots treated with native isolate both due to Fusarium moniliforme and Macrophomina phaseolina. The fungicides Carbendazim and Thiram ranked second in controlling the disease due to F. moniliforme and M. phaseolina, respectively. Plant health with respect to vigour and plant stand were found to be better than those of all the other treatments. It is concluded that the native isolate of T. harzianum may be used for maintaining good plant health and managing post-flowering stalk rot of maize.     

Comparing lignocellulose physiochemistry after decomposition by brown rot fungi with distinct evolutionary origins

Among wood‐degrading fungi, lineages holding taxa that selectively metabolize carbohydrates without significant lignin removal (brown rot) are polyphyletic, having evolved multiple times from lignin‐removing white rot fungi. Given the qualitative nature of the ‘brown rot’ classifier, we aimed to quantify and compare the temporal sequence of carbohydrate removal among brown rot clades. Lignocellulose deconstruction was compared among fungi using distinct plant substrates (angiosperm, conifer, grass). Specifically, aspen, pine and corn stalk were harvested over a 16‐week time series from microcosms containing Gloeophyllum trabeum, Fomitopsis pinicola, Ossicaulis lignatilis, Fistulina hepatica, Serpula lacrymans, Wolfiporia cocos or Dacryopinax sp. After quantifying plant mass loss, a thorough compositional analysis was complemented by a saccharification test to determine wood cell wall accessibility. Mass loss and accessibility varied depending on fungal decomposer and substrate, and trajectories of loss for hemicellulosic components and cellulose differed among plant tissue types. At any given stage of decomposition, however, lignocellulose accessibility and the fraction remaining of carbohydrates and lignin within a plant tissue type were generally the same, regardless of fungal isolate. This suggests that the sequence of plant component removal at this typical scale of characterization is shared among these brown rot lineages, despite their diverse genomes and secretomes.     

First Report of Corn Whorl Rot Caused by Serratia marcescens in China

Whorl rot is a novel disease of corn found in the Huang‐Huai‐Hai Plain, China. Common symptoms of the disease in fields include yellowing and water‐soaked brown necrosis of young leaves in the whorl of corn plants, which often results in rot of the whorl. Bacterial streaming was always observed from diseased samples. Bacterial isolates were obtained from symptomatic tissue and further confirmed to be the casual agent of the disease using Koch's Postulates. Sequence analysis of 16S rDNA, housekeeping gene groES‐groEL and BIOLOG tests revealed that the isolate B3R3 belongs to the bacterium Serratia marcescens. None of the corn cultivars evaluated showed acceptable resistance to the disease. To our knowledge, this is the first report on corn whorl rot caused by Serratia marcescens.     

Fusarium species and their mycotoxins in infected corn in Italy

Surveys of corn (infected plants and commercial kernels) forFusarium species and their mycotoxins were carried out on samples collected all over Italy and from some European and mediterranean countries.Investigations on samples of corn stalk and ear rot standing in the field, mainly collected in southern Italy, proved to be contaminated with zearalenone (ZON), zearalenols (ZOL), and deoxynivalenol (DON). TheFusarium species most frequently isolated, and their recorded toxigenic capability (in parentheses), were:F. moniliforme;F. culmorum (ZON, ZOL, DON, 3AcDON);F. equiseti (ZON, ZOL); andF. proliferatum (MF). Along with these species,F. graminearum group 2 (ZON, DON and/or 3AcDON or 15AcDON);F. chlamydosporum;F. acuminatum (type-A trichothecene derivatives); andF. semitectum were often found to be associated.F. heterosporum (ZON, ZOL);F. solani;F. crookwellense (ZON, ZOL, FUS, NIV);F. oxysporum (MF);F. avenaceum (MF);F. sporotrichioides (T-2 toxin and derivatives); andF. poae (DAS, MAS) were occasionally isolated.     

Variation in 8-ketotrichothecenes and zearalenone production by Fusarium graminearum isolates from corn and barley in Korea

A total of 214 Fusarium graminearum isolates were obtained from corn and barley which were collected from Kangwon province and the southern part of Korea, respectively, and were tested for 8-ketotrichothecenes and zearalenone (ZEA) production on rice grains. The incidences of trichothecene production by 105 isolates of F. graminearum from corn were 59.0% for deoxynivalenol (DON), 37.1% for 15-acetyldeoxynivalenol(15-ADON), 13.3% for 3-acetyldeoxynivalenol (3-ADON), 7.6% for 3,15-diacetyldeoxynivalenol (3,15-DADON), 20.0% for nivalenol (NIV), 6.7% for 4-acetylnivalenol (4-ANIV), and 1.0% for 4,15-diacetylnivalenol (4,15-DANIV). DON chemotypes frequently produced 15-ADON as the major isomer rather than 3-ADON and 9 of the 61 DON chemotypes produced low levels of NIV. On the other hand, the incidences of trichothecene production of 109 isolates by F. graminearum from barley were 24.8% for DON, 72.5% for NIV, 62.4% for 4-ANIV, and 10.1% for 4,15-DANIV. Of these isolates, 78 were NIV chemotypes and only one isolate produced DON and 3-ADON as major toxins. In addition, 26 of the 78 NIV chemotypes produced low levels of DON. ZEA was frequently produced by the trichothecene-producing isolates and the incidences of ZEA were 51.4% and 31.2% for the isolates from corn and barley, respectively. There was a great regional difference in trichothecene production by F. graminearum isolates between corn- and barley-producing areas in Korea.     

A major QTL for resistance to Gibberella stalk rot in maize

Fusarium graminearum Schwabe, the conidial form of Gibberella zeae, is the causal fungal pathogen responsible for Gibberella stalk rot of maize. Using a BC₁F₁ backcross mapping population derived from a cross between ‘1145’ (donor parent, completely resistant) and ‘Y331’ (recurrent parent, highly susceptible), two quantitative trait loci (QTLs), qRfg1 and qRfg2, conferring resistance to Gibberella stalk rot have been detected. The major QTL qRfg1 was further confirmed in the double haploid, F₂, BC₂F₁, and BC₃F₁ populations. Within a qRfg1 confidence interval, single/low-copy bacterial artificial chromosome sequences, anchored expressed sequence tags, and insertion/deletion polymorphisms, were exploited to develop 59 markers to saturate the qRfg1 region. A step by step narrowing-down strategy was adopted to pursue fine mapping of the qRfg1 locus. Recombinants within the qRfg1 region, screened from each backcross generation, were backcrossed to ‘Y331’ to produce the next backcross progenies. These progenies were individually genotyped and evaluated for resistance to Gibberella stalk rot. Significant (or no significant) difference in resistance reactions between homozygous and heterozygous genotypes in backcross progeny suggested presence (or absence) of qRfg1 in ‘1145’ donor fragments. The phenotypes were compared to sizes of donor fragments among recombinants to delimit the qRfg1 region. Sequential fine mapping of BC₄F₁ to BC₆F₁ generations enabled us to progressively refine the qRfg1 locus to a ~500-kb interval flanked by the markers SSR334 and SSR58. Meanwhile, resistance of qRfg1 to Gibberella stalk rot was also investigated in BC₃F₁ to BC₆F₁ generations. Once introgressed into the ‘Y331’ genome, the qRfg1 locus could steadily enhance the frequency of resistant plants by 32–43%. Hence, the qRfg1 locus was capable of improving maize resistance to Gibberella stalk rot.     

Tillage Effects on the Incidence and Severity of Root and Stalk Rot of Maize Caused by Fusarium moniliforme and Macrophomina phaseolina in South-Western Nigeria

Conventional tillage methods were compared with no tillage systems for the control of root and stalk rot disease of maize caused by Macrophomina phaseolina and Fusarium moniliforme in a field with a recent history of high stalk rot incidence in south-western Nigeria. The incidence of stalk rot was significantly less under no-tillage practices than in conventionally tilled plots during the twoseason trial. Application of paraquat or burning of crop residue had no apparent influence ond isease incidence. None of the treatments affected the severity of the diseases.     

Pathogenicity of<Emphasis Type="Underline">Fusarium</Emphasis><Emphasis Type="Underline">SPP</Emphasis>. contributing to the stalk rot of Maize in Poland

During 1985–1989 a stalk rot of early maturity hybrids of maize was studied in Radzikow (Central Poland). It was found thatFusarium species were dominant on plants with stalk rot symptoms. Spectrum ofFusarium spp. had changed within the years. The most frequently isolated were:F.subglutinans,F.culmorum andF.crookwellense. When the disease developed early in the seasonF.graminearum was also present.Predominant species were examined for their pathogenicity according to the modified method of Molot, Simone (1967). Isolates ofF.graminearum andF.culmorum were found to be strong pathogens,F.crookwellense andF.subglutinans — moderate andF.oxysporum andF.eguiseti were the weak ones     

Mapping quantitative trait loci (QTLs) for resistance to Gibberella zeae infection in maize

The basic prerequisite for an efficient breeding program to improve levels of resistance to pathogens in plants is the identification of genes controlling the resistance character. If the response to pathogens is under the control of a multilocus system, the utilization of molecular markers becomes essential. Stalk and ear rot caused by Gibberella zeae is a widespread disease of corn: resistance to G. zeae is quantitatively inherited. Our experimental approach to understanding the genetic basis of resistance to Gibberella is to estimate the genetic linkage between available molecular markers and the character, measured as the amount of diseased tissue 40 days after inoculation of a suspension of Fusarium graminearum, the conidial form of G. zeae, into the first stalk internode. Sensitive and resistant parental inbreds were crossed to obtain F₁ and F₂ populations: the analysis of the segregation of 95 RFLP (restriction fragment length polymorphism) clones and 10 RAPD (random amplified polymorphic DNA) markers was performed on a population of 150 F₂ individuals. Analysis of resistance was performed on the F₃ families obtained by selfing the F₂ plants. Quantitative trait loci (QTL) detection was based either on analysis of regression coefficients between family mean value and allele values in the F₂ population, or by means of interval mapping, using MAPMAKER-QTL. A linkage map of maize was obtained, in which four to five genomic regions are shown to carry factors involved in the resistance to G. zeae.     

Effect of organic soil amendments on the incidence of stalk rot of maize

Five organic materials, Calopogonium sp. leaves, rice straw, mixed wood saw dust of Terminalia sp. Triplochyton sp. and Kaya sp., fresh guinea grass and poultry manure were added to soil with a recent history of high stalk rot of maize caused by Macrophomina phaseolina and Fusarium moniliforme. The two stalk rot organisms reacted differently to the organic amendments. All the amendments produced significantly less Fusarium stalk rot disease than the unamended control. Amendment of soil with fresh Calopogonium leaves was almost ineffective in controlling the incidence of Macrophomina phaseolina. Organic amendment with poultry manure increased the disease casued by M. phaseolina. The other amendments appeared to be equally effective in checking the disease. Increased microbial population due to amendment may play some role in disease suppression.     

Reprogrammed endophytic microbial community in maize stalk induced by Trichoderma asperellum biocontrol agent against Fusarium diseases and mycotoxin accumulation

Maize stalk rot and ear rot, caused by Fusarium graminearum and Fusarium verticillioides, respectively, are major diseases that threaten the sustainable production of maize. In this study, an artificial inoculation assay demonstrated that the control efficacy of maize stalk rot and ear rot by Trichoderma asperellum granules were 49.83 % and 39.63 %, respectively. By high-throughput sequencing of maize plants, a total of 76 196 ITS1 sequences and 887 226 V3V4 16S rRNA sequences were analyzed and were grouped into 2934 fungal and 24 248 bacterial operational taxonomic units (OTUs), respectively. It revealed a significantly higher endophytic microbial abundance in the stem tissue of plants grown in T. asperellum-treated soil than in those grown in the control, with the largest increase observed in the basal stem section. In addition, the endophytic microbial diversity and corresponding control effects all gradually decreased from the basal to apical parts of the stem in plants grown in Trichoderma-treated soil, indicating that Trichoderma stimulated a more significant effect on the defense system in the basal section of the stalk than in the apical parts of plants. Furthermore, the accumulation of deoxynivalenol (DON) and fumonisin B1 (FB1) decreased in the stem and ear of maize grown in T. asperellum-treated soil.     

内生真菌印度梨形孢诱导小麦抗根腐病作用研究

【目的】明确印度梨形孢(Piriformospora indica)诱导小麦对根腐病产生抗性的作用机制。【方法】用印度梨形孢悬液浸种,以无菌培养液为对照,用病原菌禾谷镰孢菌(Fusarium graminearum)侵染小麦,对其相关生理生化指标及转录组变化进行分析。【结果】禾谷镰孢菌能诱导小麦产生过氧化氢,降低细胞内水含量,破坏细胞膜的稳定性;根部定殖印度梨形孢的小麦细胞内抗氧化酶活性增强,活性氧自由基含量降低,胞内水含量提高,细胞膜稳定性增强;印度梨形孢定殖能改变由于病原菌引起的mRNA转录组变化,抗性相关基因的表达增强。综合表明印度梨形孢定殖能有效地提高小麦对禾谷镰孢菌的抗性。【结论】研究结果为深入理解植物与微生物互作、开发新型高效环保抗根腐病生物制剂提供理论和实验依据。     

玉米感染禾谷镰刀菌后PAL、POD活性和同工酶谱的变化

为了明确苯丙氨酸解氨酶活和过氧化物酶活与玉米对镰刀菌茎腐病的抗性关系,探讨玉米对镰刀菌茎腐病的抗病机制及其在玉米抗病性鉴定中的利用,用禾谷镰刀菌孢子悬浮液对抗病品种陕单931和感病品种西农11号在抽雄初期进行接种,并于接种后测定茎秆髓部组织内的PAL,POD活性变化以及POD同工酶谱的变化。结果表明,玉米植株受镰刀菌侵染后,抗病品种的PAL酶活上升快,活性强,且形成两个活性高峰,高活性时间持续长;感病品种PAL酶活上升慢,活性相对较弱,且只形成一个峰,高活性持续时间短。抗病品种POD酶活峰值高,感病品种峰值低;抗病品种高酶活持续的时间长,感病品种高酶活持续的短。POD同工酶谱研究表明,抗、感品种POD同工酶带都有增多。抗病品种增5条,感病品种新增2条。PAL活性变化、POD活性变化及同工酶谱酶活变化与其对茎腐病的抗性有密切的关系,可作为抗病育种的生理生化指标。