Graminis Var Research Articles

Catalyst-free synthesis of novel α-trifluoromethylated tertiary alcohols bearing azaarenes as potential antifungal agents

Herein, α-trifluoromethylated tertiary alcohols bearing azaarenes were successfully synthesized via the reaction of 2-methyl azaarenes and CF3-substituted 1,3-diols with high yields under catalyst-free, mild conditions. The antifungal activities against Gaeumannomyces graminis var tritici, Rhizoctonia cerealis, Fusarium graminearum, Rhizoctonia solani, Fusarium moniliforme, Sclerotium rolfsii Sacc were investigated. The bioassay results indicated that the antifungal activity against Gaeumannomyces graminis var tritici was obviously better compared to other five fungi and compound 3dd displayed the highest activity, with an EC50 value of 27.73 μg/mL in vitro.

Associations Among Cultivar Cropping Sequence, 2,4-Diacetlyphloroglucinol-Producing Pseudomonad Populations, and Take-All Disease of Winter Wheat in Oregon.

Take-all of wheat (Triticum aestivum L.), caused by Gaeumannomyces tritici (syn. G. graminis var. tritici), is perhaps the most important soil-borne disease of wheat globally and can cause substantial yield losses under several cropping scenarios in Oregon. Though resistance to take-all has not been identified in hexaploid wheat, continuous cropping of wheat for several years can reduce take-all severity through the development of suppressive soils, a process called "take-all decline" (TAD). Extensive work has shown that TAD is driven primarily by members of the Pseudomonas fluorescens complex that produce 2,4-diacetlyphloroglucinol (DAPG), an antibiotic that is associated with antagonism and induced host resistance against multiple pathogens. Field experiments were conducted to determine the influence of agronomically relevant first year wheat cultivars on take-all levels and ability to accumulate DAPG-producing pseudomonads within their rhizospheres in second-year field trials and in greenhouse trials. One first year wheat cultivar consistently resulted in less take-all in second-year wheat and accumulated significantly more DAPG-producing pseudomonads than other cultivars, suggesting a potential mechanism for take-all reduction associated with that cultivar. An intermediate level of take-all suppression in other other cultivars was not clearly associated with population size of DAPG-producing pseudomonads, however. The first year cultivar effect on take-all dominated in subsequent plantings, and its impact was not specific to the first year cultivar. Our results confirm that wheat cultivars may be used to suppress take-all when deployed appropriately over cropping seasons, an approach that is cost effective, sustainable, and currently being utilized by some wheat growers in Oregon to reduce take-all.

Discovery of Novel Cinnamic Acid Derivatives as Fungicide Candidates.

Structural diversity derivatization from natural products is an important and effective method of discovering novel green pesticides. Cinnamic acids are abundant in plants, and their unparalleled structures endow them with various excellent biological activities. A series of novel cinnamic oxime esters were designed and synthesized to develop high antifungal agrochemicals. The antifungal activity, structure-activity relationship, and action mechanism were systematically studied. Compounds 7i, 7u, 7v, and 7x exhibited satisfactory activity against Gaeumannomyces graminis var. tritici, with inhibition rates of ≥90% at 50 μg/mL. Compounds 7z and 7n demonstrated excellent activities against Valsa mali and Botrytis cinerea, with median effective concentration (EC50) values of 0.71 and 1.41 μg/mL, respectively. Compound 7z exhibited 100% protective and curative activities against apple Valsa canker at 200 μg/mL. The control effects of 7n against gray mold on tomato fruits and leaves were all >96%, exhibiting superior or similar effects to those of the commercial fungicide boscalid. Furthermore, the quantitative structure-activity relationship was established to guide the further design of higher-activity compounds. The preliminary results on the action mechanism revealed that 7n treatment could disrupt the function of the nucleus and mitochondria, leading to reactive oxygen species accumulation and cell membrane damage. Its primary biochemical mechanism may be inhibiting fungal ergosterol biosynthesis. The novel structure, simple synthesis, and excellent activity of cinnamic oxime esters render them promising potential fungicides.

Impact of cropping and tillage system on take-all disease of winter wheat (Gaeumannomyces graminis var. tritici)

Diseases induced by Gaeumannomyces graminis var. tritici (G. g. tritici) are the most important wheat root diseases in all cropping areas around the world and significantly reduces yield and quality of wheat grain. A multi-year field experiment was used to evaluate infestation of winter wheat roots by G. g. tritici. The experimental factors included: I. cropping system (CS): 1) crop rotation (CR): pea – winter wheat – spring barley; 2) monoculture of winter wheat (MON); and II. tillage system (TS): 1) CT – conventional tillage, 2) RT – reduced tillage, and 3) NT – no-tillage. The following cultivation treatment were applied: a shallow ploughing after previous harvest and pre-sowing ploughing in the CT system; double cultivation instead of both aforementioned ploughing treatments in the RT system; and glyphosate in NT system. A cultivation set, consisting of a cultivator, a string roller, and a harrow, was used before wheat sowing. The value of the index of winter wheat roots infestation by G. g. tritici was significantly higher in the monoculture than in crop rotation. The value was also higher in NT than in CT system. In wheat monoculture with NT system, the percentage of highly and severely infested plants was higher than in crop rotation with CT system.

A novel biopolymer technique for encapsulation of Bacillus velezensis BV9 into double coating biopolymer made by in alginate and natural gums to biocontrol of wheat take-all disease

Bacillus velezensis has been known for its high potential in controlling agricultural diseases. Technological advances have opened new perspectives for producing effective formulations by reducing some of the obstacles to their use, such as instability and loss of activity due to exposure to adverse environmental conditions. Encapsulation is one of the new approaches in agricultural science. This research describes discoveries related to processes for the microencapsulation of B. velezensis with natural gums. The efficiency, survival, and controlled release of B. velesensis BV9 encapsulated with alginate mixed with zedo gum, mastic gum, and tragacanth gum were evaluated for this aim. Furthermore, under greenhouse conditions, the encapsulated cells were assessed to control Gaeumannomyces graminis var. tritici in wheat. The results indicated that all tested microcapsules protected >60 % of the bacterial cells. The Alginate-Zedo Gum (Alg-ZG) microcapsules showed a better-controlled release over two months. The greenhouse study indicated that treating wheat plants with Alg-ZG microcapsules was the most efficient treatment, suppressing 100 % of the pathogen. The results indicated that Alg-ZG is the most promising mixture to improve the survivability of B. velezensis BV9. Also, using natural gums and great potential of this formulation provides an effective and affordable fertilizers for agriculture.

1,2,4-Triazole benzamide derivative TPB against Gaeumannomyces graminis var. tritici as a novel dual-target fungicide inhibiting ergosterol synthesis and adenine nucleotide transferase function.

Isopropyl 4-(2-chloro-6-(1H-1,2,4-triazol-1-yl)benzamido)benzoate (TPB) was a 1,2,4-triazole benzoyl arylamine derivative with excellent antifungal activity, especially against Gaeumannomyces graminis var. tritici (Ggt). Its mechanism of action was investigated by transmission electron microscopy (TEM) observation, assays of sterol composition, cell membrane permeability, intracellular ATP and mitochondrial membrane potential, and mPTP permeability, ROS measurement, RNA sequencing (RNA-seq) analysis. TPB interfered with ergosterol synthesis, reducing ergosterol content, increasing toxic intermediates, and finally causing biomembrane disruption such as increasing cell membrane permeability and content leakage, and destruction of organelle membranes such as coarse endoplasmic reticulum and vacuole. Moreover, TPB destroyed the function of adenine nucleotide transferase (ANT), leading to ATP transport obstruction in mitochondria, inhibiting mPTP opening, inducing intracellular ROS accumulation and mitochondrial membrane potential loss, finally resulting in mitochondrial damage including mitochondria swelled, mitochondrial membrane dissolved, and cristae destroyed and reduced. RNA-seq analyses showed that TPB increased the expression of ERG11, ERG24, ERG6, ERG5, ERG3 and ERG2 genes in ergosterol synthesis pathway, interfered with the expression of genes (NDUFS5, ATPeV0E, NCA2 and Pam17) related to mitochondrial structure, and inhibited the expression of genes (WrbA and GST) related to anti-oxidative stress. TPB exhibited excellent antifungal activity against Ggt by inhibiting ergosterol synthesis and destroying ANT function. So, TPB was a novel compound with dual-target mechanism of action and can be considered a promising novel fungicide for the control of wheat Take-all. The results provided new guides for the structural design of active compounds and powerful tools for pathogen resistance management. © 2023 Society of Chemical Industry.

Melanin Induction Restores the Pathogenicity of Gaeumannomyces graminis var. tritici in Wheat Plants.

One of the most challenging aspects of long-term research based on microorganisms is the maintenance of isolates under ex situ conditions, particularly the conservation of phytopathological characteristics. Our research group has worked for more than 10 years with Gaumannomyces graminis var. tritici (Ggt), the main biotic factor affecting wheat. In this sense we preserved the microorganisms in oil overlaid. However, several strains preserved for a long time lost their pathogenicity. These strains show white and non-infective mycelia. In this sense, we hypothesized that this is attributable to low melanin content. Melanin is a natural pigment mainly involved in UV protection, desiccation, salinity, oxidation, and fungal pathogenicity. Therefore, understanding the melanin role on Ggt pathogenicity is fundamental to developing melanin activation strategies under laboratory studies. In this study, we induce melanin activation by UV-A light chamber, 320 to 400 nm (T1) and temperature changes of 30 °C, 15 °C, and 20 °C (T2). Fungal pathogenicity was evaluated by determination of blackening roots and Ggt was quantified by real-time PCR in inoculated wheat plants. Results revealed that Ggt grown under UV-A (T1) conditions showed around 40% higher melanin level with a concomitant effect on root infection (98% of blackened roots) and 4-fold more Ggt genome copy number compared with the control (non-infective mycelia) being T1, a more inductor factor compared with T2. These findings would support the role of melanin in pathogenicity in darkly pigmented fungi such as Ggt and could serve as a basis for activating pathogenicity under laboratory conditions.

Management of take-all disease caused by Gaeumannomyces graminis var. tritici in wheat through Bacillus subtilis strains.

Wheat (Triticum aestivum) is the second largest grain crop worldwide, and one of the three major grain crops produced in China. Take-all disease, caused by Gaeumannomyces graminis var. tritici (Ggt) infection, is a widespread and devastating soil-borne disease that harms wheat production. At present, the prevention and control of wheat take-all depend largely on the application of chemical pesticides. Chemical pesticides, however, not only lead to increased drug resistance of pathogens but also leave significant residues in the soil, causing serious environmental pollution. In this study, we investigated the application of Bacillus subtilis to achieve take-all disease control in wheat while reducing pesticide application. Antagonistic bacteria were screened by plate test, species identification of strains was performed by Gram staining and sequencing of 16s rDNA, secondary metabolite activity of strains was detected by clear circle method, strain compatibility and effect of compounding on Ggt were detected by plate, and the application prospects of specific strains were analyzed by greenhouse and field experiments. We found that five B. subtilis strains, JY122, JY214, ZY133, NW03, Z-14, had significant antagonistic effects against Ggt, and could secrete antimicrobial proteins including amylase, protease, and cellulase. Furthermore, Z-14 and JY214 cultures have also been shown to change the morphology of Ggt mycelium. These results also showed that Z-14, JY214, and their combination can control take-all disease in wheat at a reduced level of pesticide use. In summary, we screened two Bacillus spp. strains, Z-14 and JY214, that could act as antagonists that contribute to the biological control of wheat take-all disease. These findings provide resources and ideas for controlling crop diseases in an environmentally friendly manner.

Potential Biocontrol Activities of Populus Endophytes against Several Plant Pathogens Using Different Inhibitory Mechanisms

The plant microbiome can be used to bolster plant defense against abiotic and biotic stresses. Some strains of endophytes, the microorganisms within plants, can directly inhibit the growth of plant fungal pathogens. A previously isolated endophyte from wild Populus (poplar), WPB of the species Burkholderia vietnamiensis, had robust in vitro antifungal activity against pathogen strains that are highly virulent and of concern to Pacific Northwest agriculture: Rhizoctonia solani AG-8, Fusarium culmorum 70110023, and Gaemannomyces graminis var. tritici (Ggt) ARS-A1, as well as activity against the oomycete, Pythium ultimum 217. A direct screening method was developed for isolation of additional anti-fungal endophytes from wild poplar extracts. By challenging pathogens directly with dilute extracts, eleven isolates were found to be inhibitory to at least two plant pathogen strains and were therefore chosen for further characterization. Genomic analysis was conducted to determine if these endophyte strains harbored genes known to be involved in antimicrobial activities. The newly isolated Bacillus strains had gene clusters for production of bacillomycin, fengicyn, and bacillibactin, while the gene cluster for the synthesis of sessilin, viscosin and tolaasin were found in the Pseudomonas strains. The biosynthesis gene cluster for occidiofungin (ocf) was present in the Burkholderia vietnamiensis WPB genome, and an ocf deletion mutant lost inhibitory activity against 3 of the 4 pathogens. The new isolates lacked the gene cluster for occidiofungin implying they employ different modes of action. Other symbiotic traits including nitrogen fixation, phosphate solubilization, and the production of auxins and siderophores were investigated. Although it will be necessary to conduct in vivo tests of the candidates with pathogen-infected agricultural crops, the wild poplar tree microbiome may be a rich source of beneficial endophyte strains with potential for biocontrol applications against a variety of pathogens and utilizing varying modes of action.

Influence of Bacillus subtilis strain Z-14 on microbial communities of wheat rhizospheric soil infested with Gaeumannomyces graminis var. tritici.

Wheat take-all disease caused by Gaeumannomyces graminis var. tritici (Ggt) spreads rapidly and is highly destructive, causing severe reductions in wheat yield. Bacillus subtilis strain Z-14 that significantly controlled wheat take-all disease effectively colonized the roots of wheat seedlings. Z-14 increased the metabolic activity and carbon source utilization of rhizospheric microorganisms, thus elevating average well-color development (AWCD) values and functional diversity indexes of soil microbial communities. Z-14 increased the abundance of Bacillus in the rhizosphere, which was positively correlated with AWCD and functional diversity indexes. The Z-14-treated samples acquired more linkages and relative connections between bacterial communities according to co-occurrence network analyses. After the application of Ggt, the number of linkages between fungal communities increased but later decreased, whereas Z-14 increased such interactions. Whole-genome sequencing uncovered 113 functional genes related to Z-14’s colonization ability and 10 secondary metabolite gene clusters in the strain, of which nine substances have antimicrobial activity. This study clarifies how bacterial agents like Z-14 act against phytopathogenic fungi and lays a foundation for the effective application of biocontrol agents.

Structure-Activity Studies of N-Heterocyclic Benzoyl Arylamine Derivatives Led to a Highly Fungicidal Candidate against Gaeumannomyces graminis var. tritici and Four Fusarium Wheat Pathogens.

Wheat root diseases can seriously reduce yields and quality of wheat. 1,2,4-Triazole benzoyl arylamine derivatives previously showed good activities against some wheat root fungal pathogens. To further systematically disclose the structure-activity relationship, a series of benzoyl arylamines were designed and prepared. Their structures were characterized and fungicidal activities against Gaeumannomyces graminis var. tritici and Fusarium graminearum were evaluated. The results indicated that the structure of the N-heterocyclic group and the substituted group and their position on the benzamide scaffold had an important influence on the activities, as predicted. Finally, compound 18f was found to show excellent activities against G. graminis var. tritici, F. graminearum, Fusarium culmorum, Fusarium pseudograminearum, and Fusarium moniliforme with half-maximum effective concentrations of 0.002, 0.093, 0.011, 0.881, and 0.287 μg/mL, respectively. These results proposed that compound 18f deserved serious consideration as a novel fungicide candidate for the control of wheat root diseases.

Effects of dryland summer cover crops and a weedy fallow on soil water, disease levels, wheat growth and grain yield in a Mediterranean-type environment

Numerous studies have reported wide-ranging benefits of cultivating cover crops during the typical fallow period in temperate cropping systems. However, data on the impacts of summer cover crops in water-limited, Mediterranean-type climates, is lacking. A 3-year field experiment was conducted near the town of Northam in the wheat-belt of Western Australia to test the hypothesis that inclusion of summer cover crops would increase ground cover, water storage and grain yield in subsequent wheat crops. White French millet ( Panicum miliaceum ) and cowpea ( Vigna unguiculata ) cover crops and a weedy fallow were compared to a typical chemical fallow over the summer period. Autumn groundcover was 10–30% higher when summer weeds were left uncontrolled compared to the chemical fallow treatment. Dry-sown and late established (February) millet failed to increase groundcover, whereas cowpea demonstrated a small increase (~9–10%) in one season, when cowpea biomass approached 1 t ha −1 . Cover crops and weeds growing over summer had no significant effect on total plant available water (0–50 cm depth) prior to sowing wheat in any season. However, there was less readily available soil N in the cover crop (less 10–30 kg N ha −1 ) and weedy fallow (less 50 kg N ha −1 ) treatments prior to the second wheat crop. Despite some significant effects of weedy fallow and cowpeas on Gaeumannomyces graminis var tritici (take-all) and Didymella pinodes (causing black pea spot), disease levels were low and thought to pose little risk to following winter crops. Wheat grain yields were not significantly different between treatments, other than a low yield after cowpeas in 2015 due to a low wheat emergence count. Grain protein and screenings were not generally affected by summer fallow treatment in any year, except for higher screenings after the weedy fallow in one year. Given the limited impacts of cover crops and weeds on soil water and disease, it is possible that summer cover crops may be able to be integrated with cereal winter rotations in Western Australia without yield reductions in wheat crops. In light of current chemical usage, issues around herbicide residues in soil and herbicide-resistant weeds, it would be worthwhile revisiting the need for chemical fallow, particularly in the higher rainfall areas.

Identification, frequency of occurrence, and inoculum density of select ectotrophic root‐infecting fungi within ultradwarf hybrid bermudagrass greens in Mississippi

AbstractGaeumannomyces graminis var. graminis (Sacc.) Arx & Oliver was described as a pathogen of bermudagrass decline [takeall root rot (TARR)]; however, recent studies have reported that multiple ectotrophic root‐infecting (ERI) fungi are associated with root rot in ultradwarf hybrid bermudagrass (UDHB) [Cynodon dactylon (L.) Pers. × Cynodon transvaalensis Burtt Davy] putting greens. This study surveyed UDHB roots from a healthy and a TARR‐symptomatic green. Real‐time polymerase chain reaction multiplex assays facilitated the rapid identification and quantification of four ERI fungi: G. graminis, G. nanograminis P.L Vines & M. Tom.‐Pet., Magnaporthiopsis cynodontis P.L Vines & M. Tom.‐Pet., and Candidacolonium cynodontis P.L Vines & M. Tom.‐Pet. Gaeumannomyces nanograminis was identified more frequently than G. graminis in roots from the healthy green and had the greatest inoculum density in 2018. In contrast, G. graminis was identified in 80% more root samples than G. nanograminis in the TARR‐symptomatic green in 2017, and the inoculum density of G. graminis was >2 M DNA copies per 2‐µl DNA sample. Magnaporthiopsis cynodontis was the predominant fungus in both greens throughout the study; however, its inoculum density was the lowest among all four ERI fungi. Two or more ERI fungi were frequently identified as co‐colonizing UDHB roots. This study showed that multiple ERI pathogens were widely distributed throughout the roots of two UDHB greens regardless of symptom expression and confirmed that ERI complexes co‐colonize UDHB roots. This is the first study documenting the widespread occurrence and inoculum density levels of multiple ERI fungi colonizing UDHB roots.

Delving into defence: identifying the Pseudomonas protegens Pf-5 gene suite involved in defence against secreted products of fungal, oomycete and bacterial rhizosphere competitors.

Competitive behaviours of plant growth promoting rhizobacteria (PGPR) are integral to their ability to colonize and persist on plant roots and outcompete phytopathogenic fungi, oomycetes and bacteria. PGPR engage in a range of antagonistic behaviours that have been studied in detail, such as the production and secretion of compounds inhibitory to other microbes. In contrast, their defensive activities that enable them to tolerate exposure to inhibitory compounds produced by their neighbours are less well understood. In this study, the genes involved in the Pseudomonas protegens Pf-5 response to metabolites from eight diverse rhizosphere competitor organisms, Fusarium oxysporum, Rhizoctonia solani, Gaeumannomyces graminis var. tritici, Pythium spinosum, Bacillus subtilis QST713, Pseudomonas sp. Q2-87, Streptomyces griseus and Streptomyces bikiniensis subspecies bikiniensi, were examined. Proximity induced excreted metabolite responses were confirmed for Pf-5 with all partner organisms through HPLC before culturing a dense Pf-5 transposon mutant library adjacent to each of these microbes. This was followed by transposon-directed insertion site sequencing (TraDIS), which identified genes that influence Pf-5 fitness during these competitive interactions. A set of 148 genes was identified that were associated with increased fitness during competition, including cell surface modification, electron transport, nucleotide metabolism, as well as regulatory genes. In addition, 51 genes were identified for which loss of function resulted in fitness gains during competition. These included genes involved in flagella biosynthesis and cell division. Considerable overlap was observed in the set of genes observed to provide a fitness benefit during competition with all eight test organisms, indicating commonalities in the competitive response to phylogenetically diverse micro-organisms and providing new insight into competitive processes likely to take place in the rhizosphere.

Efficient Biocontrol of Gaeumannomyces graminis var. Tritici in Wheat: Using Bacteria Isolated from Suppressive Soils

“Take-all” disease is the most important biotic factor affecting cereal productivity, causing 30–50% of crop losses. The causal agent is the ascomycete soil-borne pathogen Gaeumannomyces graminis var. tritici (Ggt). Current control measures are ineffective, because Ggt can remain saprophytic in soils for long periods. Therefore, the study of the microbiome residing in suppressive soils (SS) is a promising niche of Ggt biocontrol. Here, we evaluated the efficiency of Serratia sp., Bacillus sp., and Acinetobacter sp. isolated from SS against the incidence of Ggt on wheat. Our results demonstrated that plants inoculated with the bacterial consortium in both greenhouse and field conditions were highly efficient in Ggt biocontrol, more so than individual strains. The disease reduction was evidenced by higher biomass production, fewer copies of the Ggt genome with a concomitant curtailment of blackening of roots, a decrease of lipid peroxidation, and an increase of superoxide dismutase activity. The ability of the microbial consortium over that of single strains could be attributable to interspecies communication as a strategy to biocontrol; i.e., higher chitinase activity. In conclusion, bacterial consortia from SS are an important niche of Ggt biocontrol, serving as a model for other soil-borne pathogens.

4-Ethylphenol, A Volatile Organic Compound Produced by Disease-Resistant Soybean, Is a Potential Botanical Agrochemical Against Oomycetes.

Oomycetes, represented by Phytophthora, are seriously harmful to agricultural production, resulting in a decline in grain quality and agricultural products and causing great economic losses. Integrated management of oomycete diseases is becoming more challenging, and plant derivatives represent effective alternatives to synthetic chemicals as novel crop protection solutions. Biologically active secondary metabolites are rapidly synthesized and released by plants in response to biotic stress caused by herbivores or insects, as well as pathogens. In this study, we identified groups of volatile organic compounds (VOCs) from soybean plants inoculated with Phytophthora sojae, the causal agent of soybean root rot. 4-Ethylphenol was present among the identified VOCs and was induced in the incompatible interaction between the plants and the pathogen. 4-Ethylphenol inhibited the growth of P. sojae and Phytophthora nicotianae and had toxicity to sporangia formation and zoospore germination by destroying the pathogen cell membrane; it had a good control effect on soybean root rot and tobacco black shank in the safe concentration range. Furthermore, 4-Ethylphenol had a potent antifungal activity against three soil-borne phytopathogenic fungi, Rhizoctonia solani, Fusarium graminearum, and Gaeumannomyces graminis var tritici, and four forma specialis of Fusarium oxysporum, which suggest a potential to be an eco-friendly biological control agent.

Biological Control of Take-All and Growth Promotion in Wheat by Pseudomonas chlororaphis YB-10.

Wheat is a worldwide staple food crop, and take-all caused by Gaeumannomyces graminis var. tritici can lead to a tremendous decrease in wheat yield and quality. In this study, strain YB-10 was isolated from wheat rhizospheric soil and identified as Pseudomonas chlororaphis by morphology and 16S rRNA gene sequencing. Pseudomonas chlororaphis YB-10 had extracellular protease and cellulase activities and strongly inhibited the mycelium growth of Gaeumannomyces graminis var. tritici in dual cultures. Up to 87% efficacy of Pseudomonas chlororaphis YB-10 in controlling the take-all of seedlings was observed in pot experiments when wheat seed was coated with the bacterium. Pseudomonas chlororaphis YB-10 was also positive for indole acetic acid (IAA) and siderophore production, and coating wheat seed with the bacterium significantly promoted the growth of seedlings at 107 and 108 CFU/mL. Furthermore, treatment with Pseudomonas chlororaphis YB-10 increased activities of the wheat defense-related enzymes POD, SOD, CAT, PAL and PPO in seedlings, indicating induced resistance against pathogens. Overall, Pseudomonas chlororaphis YB-10 is a promising new seed-coating agent to both promote wheat growth and suppress take-all.

First Report of Fusarium culmorum and Microdochium bolleyi Causing Root Rot on Triticale in Kazakhstan.

Triticale (×Triticosecale Wittmack) is obtained from wheat × rye crossing. It is positioned between wheat and rye in terms of resistance to soilborne pathogens including Gaeumannomyces graminis var. tritici, Fusarium culmorum, F. avenaceum, and Bipolaris sorokiniana (Arseniuk and Góral 2015). In 2019, seven triticale fields were surveyed in Almaty Province, Kazakhstan to examine soil-borne fungal pathogens. A total of 28 symptomatic plants with stunting, rot or discolored root were collected to identify causal agents. The overall disease incidence was approximately 8 to 10% in the fields. Fungi were isolated from 3-5 mm pieces excised from symptomatic tissues. The pieces were exposed to surface disinfection in 1% sodium hypochlorite solution for 2 min, rinsed three times with sterile distilled water, blotted dry, and plated on 1/5 strength potato dextrose agar (PDA) amended with 0.01% streptomycin. Plates were left in the dark at 23°C for 7 days. A total of 34 fungal colonies were isolated of which nineteen isolates, originally from six fields showed the cultural characteristics of B. sorokiniana. This species was previously reported to cause common root rot on triticale in Kazakhstan (Özer et al. 2020). Ten isolates from four fields produced pale orange and cottony mycelium with red pigmentation on the agar, which is typical of Fusarium-like growth. The remaining isolates (n=5) from two fields produced salmon-colored and scarce aerial mycelium with no soluble pigmentation, similar to Microdochium spp. Fusarium isolates produced thick-walled and curved macroconidia with 3-4 septa (n=50, 25.7 to 37.6 × 4.1 to 7.3 μm in size) and notched basal cell on PDA, but microconidia were absent, which matches the description of F. culmorum (Wm.G. Sm.) Sacc. (Leslie and Summerell 2006). Microdochium isolates produced swollen, brown, and thick-walled chlamydospores and hyaline, one-celled, and thin-walled conidia (n=50, 5.4 to 9.3 × 1.5 to 3.0 μm in size) formed on ampullate and cylindrical conidiogenous cells on oatmeal agar (OA). These morphological features are consistent with previous observations for Microdochium bolleyi (R. Sprague) de Hoog & Herm.-Nijh. (Hong et al. 2008). To confirm morphological preliminary identifications, the portion of the translation elongation factor 1-alpha (EF1-α) gene was amplified with EF1/EF2 primers (O'Donnell et al. 1998) for representative Fusarium isolates (n=4) for each field. Additionally, the internal transcribed spacer (ITS) of ribosomal DNA was amplified with ITS1/ITS4 primers (White et al. 1990) for representative Microdochium isolates (n=2) for each field. BLASTn queries against NCBI GenBank revealed that the EF1-α sequences of Fusarium isolates (MW311081-MW311084) shared 100% identity with F. culmorum strain CBS 110262 (KT008433). The ITS sequences of M. bolleyi isolates (MW301448-MW301449) matched that of M. bolleyi strain CBS 137.64 (AM502264) with 100% sequence similarity. Pathogenicity test was conducted on pregerminated seeds of triticale cv. Balausa. A plastic pot (17 cm height, 9 cm in diam) was filled with a sterile mixture of vermiculite, peat, and soil (1:1:1, v/v/v). Mycelial plugs (1 cm in diam) were cut from the margin of a growing culture of representative isolates (Kaz_Fus123 and Kaz_Mb01) and placed onto the mixture in the pot. A sterile agar plug was employed as a control treatment. One pregerminated seed was put on the plug and covered with the mixture. The pots were transferred to a growth chamber set at 23 ± 2°C and a photoperiod of 14 hours. The experiment was performed twice using 5 replication pots per isolate. Four weeks after inoculation, discoloration of the crown was observed on all the inoculated roots, whereas no symptoms were observed on the control plants. Koch's postulates were fulfilled by reisolating and identifying the pathogen based on the morphology described above. This is the first report of M. bolleyi and F. culmorum causing root rot on triticale in Kazakhstan. Although B. sorokiniana is the most primary pathogen that may limit yield in the production of triticale in Kazakhstan, F. culmorum and M. bolleyi have been found to be less frequent and less aggressive pathogens, respectively. Further studies are needed to better understand the potential distribution and impact of these pathogens on triticale.

Activity of Fengycin and Iturin A Isolated From Bacillus subtilis Z-14 on Gaeumannomyces graminis Var. tritici and Soil Microbial Diversity.

Bacillus subtilis Z-14 can inhibit phytopathogenic fungi, and is used as a biocontrol agent for wheat take-all disease. The present study used the soil-borne fungus Gaeumannomyces graminis var. tritici (Ggt), which causes wheat take-all disease, and the soil microbial community as indicators, and investigated the antifungal effects of fengycin and iturin A purified from strain Z-14 using high performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, respectively. The results showed that fengycin destroyed the internal structure of Ggt cells by digesting the cytoplasm and organelles, forming vacuoles, and inducing hyphal shrinkage and distortion. Iturin A induced cell wall disappearance, membrane degeneration, intracellular material shrinkage, and hyphal fragmentation. A biocontrol test demonstrated a 100% control effect on wheat take-all when wheat seedlings were treated with fengycin at 100 μg/ml or iturin A at 500 μg/ml. Iturin A and fengycin both reduced the relative abundance of Aspergillus and Gibberella. At the genus level, iturin A reduced the relative abundance of Mortierella and Myrothecium, while fengycin reduced that of Fusarium. Only fengycin treatment for 7 days had a significant effect on soil bacterial diversity.

Anti-phytopathogen terpenoid glycosides from the root bark of Chytranthus macrobotrys and Radlkofera calodendron

Chytranthus macrobotrys and Radlkofera calodendron are two Sapindaceae characterized by a lack of phytochemical data. Both root barks from the two Sapindaceae species were processed by ethanol extraction followed by the isolation of their primary constituents by liquid chromatography. This process yielded four previously undescribed terpenoid glycosides together with eight known analogues. Extracts and isolated compounds from C. macrobotrys and R. calodendron were then screened for antimicrobial activity against fifteen phytopathogens. The biological screening also involved extracts and pure compounds from Blighia unijugata and Blighia welwitschii, two Sapindaceae previously studied by our group. Phytopathogens were chosen based on their economic impact on agriculture worldwide. The selection was composed primarily of fungal species including; Pyricularia oryzae, Gaeumannomyces graminis var. tritici, Zymoseptoria tritici, Fusarium oxysporum, Botrytis cinerea, Pythium spp., Trichoderma spp. and Rhizoctonia solani. Furthermore, pure terpenoid glycosides were tested for the first time against wood-inhabiting phytopathogens such as; Phaeomoniella chlamydospora, Phaeoacremonium minimum, Fomitiporia mediterranea, Eutype lata and Xylella fastidiosa. Raw extracts exhibited different levels of activity dependent on the organism. Some pure compounds, including 3-O-α-L-arabinopyranosyl-(1 → 4)-β-D-xylopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin (α-hederin), 3-O-β-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin (macranthoside A) and 3-O-α-L-arabinopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin (clemontanoside C), exhibited significant growth inhibitions on Pyricularia oryzae, Gaeumannomyces graminis var. tritici, Fomitiporia mediterranea and Zymoseptoria tritici. Monodesmoside triterpene saponins, in particular, exhibited MIC (IC100) values as low as 25 μg/ml and IC50 values as low as 10 μg/ml against these phytopathogens. Structure-activity relationships, as well as plant-microbe interactions, were discussed.