Seedling Blight Research Articles

Characterization of differences in seed endophytic microbiome in conventional and organic rice by amplicon-based sequencing and culturing methods.

In this paper, we studied the differences in the endophytic microbial composition of rice seeds grown in conventional and organic farming systems. Our results demonstrate a greater bacterial diversity in conventional farming, while organic farming showcases a higher fungal diversity. Additionally, our research reveals the ability of seed bacterial endophytes to inhibit the growth of three fungal pathogens responsible for causing seedling blight in rice. This study provides valuable insights into the potential use of beneficial seed microbial endophytes for developing a novel microbiome-based strategy in the management of rice diseases. Such an approach has the potential to enhance overall plant health and improve crop productivity.

Impact of Quorum Sensing on the Virulence and Survival Traits of Burkholderia plantarii.

Quorum sensing (QS) is a mechanism by which bacteria detect and respond to cell density, regulating collective behaviors. Burkholderia plantarii, the causal agent of rice seedling blight, employs the LuxIR-type QS system, common among Gram-negative bacteria, where LuxI-type synthase produces QS signals recognized by LuxR-type regulators to control gene expression. This study aimed to elucidate the QS mechanism in B. plantarii KACC18965. Through whole-genome analysis and autoinducer assays, the plaI gene, responsible for QS signal production, was identified. Motility assays confirmed that C8-homoserine lactone (C8-HSL) serves as the QS signal. Physiological experiments revealed that the QS-defective mutant exhibited reduced virulence, impaired swarming motility, and delayed biofilm formation compared to the wild type. Additionally, the QS mutant demonstrated weakened antibacterial activity against Escherichia coli and decreased phosphate solubilization. These findings indicate that QS in B. plantarii significantly influences various pathogenicity and survival traits, including motility, biofilm formation, antibacterial activity, and nutrient acquisition, highlighting the critical role of QS in pathogen virulence and adaptability.

Biocontrol of fungal pathogens and growth promotion in the Korean fir (Abies koreana E.H.Wilson) seedling using Bacillus velezensis CE 100

The restoration of the endangered Korean fir (Abies koreana E.H.Wilson) forests requires mass production of high-quality and well-developed seedlings for outplanting. However, phytopathogenic fungal infections lower the survival rate and growth vigor of A. koreana seedlings, frustrating the re-afforestation efforts. This study isolated and identified the phytopathogenic fungi causing seedling blight and wilt diseases in A. koreana seedlings and investigated the potential of Bacillus velezensis CE 100 in controlling the fungal pathogens and promoting seedling growth. Phomopsis mali and Fusarium oxysporum were confirmed as the causal agents of blight and wilt diseases, respectively, and both fungi reduced the survival rate of A. koreana seedlings. B. velezensis CE 100 produced cell wall-degrading enzymes: chitinase, protease, and β-1,3-glucanase, and the bacterial crude enzyme fraction inhibited the spore germination and mycelial growth of P. mali and F. oxysporum in a concentration-dependent manner, causing severe morphological deformations. Consequently, treatment with B. velezensis CE 100 improved the survival rate of seedlings infected with either P. mali or F. oxysporum compared to the control and PB media treatment. The bacterium also increased nutrient availability through ammonia–nitrogen production and phosphate solubilization and produced indole-3-acetic acid (IAA), and its inoculation remarkably improved seedling growth (root and shoot dry weight) compared to the control and PB media treatment group. This is the first study to report P. mali and F. oxysporum as the pathogens causing seedling blight and wilt diseases in A. koreana nursery seedling, and the possibility of using B. velezensis CE 100 to control the fungal pathogens and improve the seedling growth without chemical use.

Development of a qPCR Assay for the Detection and Quantification of the Fungal Pathogen Calonectria canadiana on Conifers

ABSTRACTA real‐time PCR TaqMan assay was developed for the detection of Calonectria canadiana, a fungal pathogen responsible for damping off, root rot and seedling blight in conifer forest nurseries in central and eastern North America. While highly significant in Quebec Forest nurseries, coniferous seedling mortality decreased significantly when nurseries transitioned from bare root to container seedling production. However, over the past few years, this pathogen has re‐emerged as a threat and millions of container white spruce seedlings were culled in two nurseries in eastern Quebec. A sensitive detection and quantification assay for C. canadiana was essential to investigate the biological and environmental factors driving this new epidemic. We designed primers and a TaqMan probe targeting the internal transcribed spacer (ITS) of C. canadiana. The resulting Ccan TaqMan assay successfully differentiated C. canadiana from other soil‐borne pathogens of the Nectriaceae encountered in Quebec Forest nurseries. The limit of detection of the assay was established at eight copies of C. canadiana ITS. The Ccan TaqMan assay quickly identified the presence of the pathogen in both symptomatic and asymptomatic white spruce (Picea glauca) seedlings. Furthermore, we demonstrated that the pathogen was more easily detected when DNA was extracted from necrotic needles at the base of the stem rather than from necrotic roots. This molecular tool will greatly aid in understanding the biology and epidemiology of C. canadiana.

Streptomyces pratensis S10 Promotes Wheat Plant Growth and Induces Resistance in Wheat Seedlings against Fusarium graminearum.

Fusarium graminearum, a devastating fungal pathogen, causes great economic losses to crop yields worldwide. The present study investigated the potential of Streptomyces pratensis S10 to alleviate F. graminearum stress in wheat seedlings based on plant growth-promoting and resistance-inducing assays. The bioassays revealed that S10 exhibited multiple plant growth-promoting properties, including the production of siderophores, 1-aminocyclopropane-1-carboxylic acid deaminase (ACC), and indole-3-acetic acid (IAA), phosphate solubilization, and nitrogen fixation. Meanwhile, the pot experiment demonstrated that S10 improved wheat plant development, substantially enhancing wheat height, weight, root activity, and chlorophyll content. Consistently, genome mining identified abundant genes associated with plant growth promotion. S10 induced resistance against F. graminearum in wheat seedlings. The disease incidence and disease index reduced by nearly 52% and 65% in S10 pretreated wheat seedlings, respectively, compared with those infected with F. graminearum only in the non-contact inoculation assay. Moreover, S10 enhanced callose deposition and reactive oxygen species (ROS) accumulation and induced the activities of CAT, SOD, POD, PAL, and PPO. Furthermore, the quantitative real-time PCR (qRT-PCR) results indicated that S10 pretreatment increased the expression of SA- (PR1.1, PR2, PR5, and PAL1) and JA/ET-related genes (PR3, PR4a, PR9, and PDF1.2) in wheat seedlings upon F. graminearum infection. In summary, S. pratensis S10 could be an integrated biological agent and biofertilizer in wheat seedling blight management and plant productivity enhancement.

A Large-Scale Hydroponic Evaluation of Rice Mutants for Pythium Resistance.

Rice is a major staple crop worldwide. However, the occurrence of rice diseases during cultivation poses a significant challenge to achieving optimal yields. Among the major pathogens, Pythium species, which cause seedling blight, are of particular concern. Pythium infects rice roots through zoospores, mycelia, or oospores, leading to root rot, stunting, yellowing, and ultimately seedling damping-off. While many disease resistance-related genes have been reported in rice, only very limited research has been associated with resistance to Pythium infection. In this study, we aimed to establish a rapid screening system to identify rice lines that are resistant or susceptible to the Pythium pathogen in rice nurseries. We conducted evaluations on important factors, including virulence, inoculation method, seed-soaking period, and the measurement of disease severity. As a result, we successfully developed a screening system that allows for high-throughput and rapid screening of the Taiwan Rice Insertional Mutant (TRIM) library for mutant lines exhibiting resistance to P. arrhenomanes. Furthermore, we identified a slightly resistant TRIM line and explored potential genes encoding endoglucanase-1 precursor and malonyl-CoA decarboxylase that may be involved in conferring resistance to P. arrhenomanes.

Direct PCR Based Detection of Macrophomina Phaseolina From Soil of Jute Field

Macrophomina phaseolina (Tassi) Goid. is a devastating fungal pathogen which infects a large number of economically important crops. In jute, it mainly causes seedling blight, leaf spot, and stem rot. The pathogen is seed borne as well as soil borne. Macrophomina phaseolina pathogen was detected in soil samples collected from jute field by a direct PCR method (dPCR) without DNA isolation using ITS2 and ITS3 region specific primers designed during the present study. The soil samples were prepared in a lysis buffer and were incubated at 650C for 15 min. The optimized buffer system was composed of 200 mM tris (hydroxymethyl aminomethane (Tris)-Cl (pH 8.0), 3.5 mM ethylene diamine tetra acetic (EDTA) (pH 8.0), 4 M NaCl and 200 μg/ml Proteinase K. Further, β-marcaptoethanol (1% v/v) was added into the buffer. Bangladesh J. Bot. 53(2): 401-404, 2024 (June)

Effect of abscisic acid on rice defense mechanism against Fusarium oxysporum

Fusarium oxysporum is one of the most destructive pathogens which causes rice seedling blight. ABA is part of a large signaling system that provides an effective system against microbial and environmental manipulations. The role of ABA in plant defense mechanisms is not clear. In this experiment, we prove the role of abscisic acid (ABA) in boosting rice plant resistance against F. oxysporum and optimizing ABA concentrations against F. oxysporum. This study is divided into two experiments. In the first experiment, we used various ABA concentrations of 0.0, 0.05, 0.1, 0.2, and 0.25 mmol/L under F. oxysporum stress. In the second experiment, we use Fluridone FLD as an ABA inhibitor with the following treatments, (F) is only applied with F. oxysporum (ABA+F), abscisic acid with F. oxysporum (ABAI+F), ABA inhibitor Fluridone with F. oxysporum (ABAI), where only ABA inhibitor Fluridone was applied and CK was used as a control. The results revealed that all the plants treated with ABA exhibit better performance against F. oxysporum, except those treated without ABA. ABA concentrations of 0.2 mmol/L effectively decreased the disease index and disease incidence rate as well as improved the quality of seedlings. ABA effectively increased the activity of defense-related enzymes like PPO, POD, PAL and SOD. ABA also lowers down the MDA content which proves its effectiveness against F. oxysporum. ABA resistance was also proved by plants treated with the abscisic acid inhibitor ABAI (Fluridone FLD). The ABA inhibitor reduced the rice resistance to F. oxysporum, by conforming the expression of defense-related genes PRB1-3, PRBI-2 and Xa39(t). These gene expressions indicate the involvement of ABA in plant defense system.

Data-Driven Characterization of Metabolome Reprogramming during Early Development of Sorghum Seedlings.

Specialized metabolites are produced via discrete metabolic pathways. These small molecules play significant roles in plant growth and development, as well as defense against environmental stresses. These include damping off or seedling blight at a post-emergence stage. Targeted metabolomics was followed to gain insights into metabolome changes characteristic of different developmental stages of sorghum seedlings. Metabolites were extracted from leaves at seven time points post-germination and analyzed using ultra-high performance liquid chromatography coupled to mass spectrometry. Multivariate statistical analysis combined with chemometric tools, such as principal component analysis, hierarchical clustering analysis, and orthogonal partial least squares-discriminant analysis, were applied for data exploration and to reduce data dimensionality as well as for the selection of potential discriminant biomarkers. Changes in metabolome patterns of the seedlings were analyzed in the early, middle, and late stages of growth (7, 14, and 29 days post-germination). The metabolite classes were amino acids, organic acids, lipids, cyanogenic glycosides, hormones, hydroxycinnamic acid derivatives, and flavonoids, with the latter representing the largest class of metabolites. In general, the metabolite content showed an increase with the progression of the plant growth stages. Most of the differential metabolites were derived from tryptophan and phenylalanine, which contribute to innate immune defenses as well as growth. Quantitative analysis identified a correlation of apigenin flavone derivatives with growth stage. Data-driven investigations of these metabolomes provided new insights into the developmental dynamics that occur in seedlings to limit post-germination mortality.

Biological Control Ability and Antifungal Activities of Bacillus velezensis Bv S3 against Fusarium oxysporum That Causes Rice Seedling Blight

Fusarium oxysporum, a primary soil-borne fungus that affects rice seedlings globally, is responsible for rice seedling blight (RSB), which reduces seedling quality and survival rates. The synthetic fungicides used to treat this disease negatively affect human health and the environment. A biocontrol bacterial isolate, Bacillus velezensis Bv S3, isolated from the rice rhizosphere, showed a strong antagonistic effect on RSB-causing F. oxysporum. The ratio of the longest to the shortest radius of F. oxysporum following Bv S3 inoculation was 2.52 (cm/cm) in a plate standoff experiment. This was different from the other biocontrol strains. Bv S3 exhibits a wide spectrum of antifungal activity against various pathogenic fungi that cause RSB. When 10% Bv S3 liquid culture filtrate was applied, it dramatically reduced F. oxysporum spore germination and mycelial growth, with inhibition rates of 66.7%, and 45.7%, respectively, and caused hyphal malformations. Furthermore, the Bv S3 suspension (1 × 108 CFU/mL) reduced RSB by 65.5% and 76.5% in pot experiments, effectively promoted the growth of rice seedlings, and improved the activities of neutral phosphatase, urease, invertase, and catalase in rice rhizosphere soil. The active substances produced by Bv S3 were sensitive to temperature and ultraviolet irradiation, and the antifungal effect significantly increased after 90 min of exposure, with antifungal effect observed at pH 7. Bv. S3 effectively reduced the incidence of RSB and showed potential as a biocontrol agent.

Biopriming with Trichoderma harzianum: an effective tool for management of soil borne diseases of safflower

An effect of biopriming with Trichoderma harzianum on different soil borne diseases of safflower was studied at three consecutive year 2018-19, 2019-20 and 2020-21 using safflower variety safflower variety PBNS-12. Safflower seed biopriming with Trichoderma harzianum @ 10g/litre water for 12 hrs (10 gm Thz + 1 ltr water + 1 kg seed + 12 hrs soaking) before sowing was found very effective and economical management of Phytophthora seedling blight, Fusarium wilt and Rhizoctonia root rot diseases of Safflower.

Isolation and Cultural, Morphological Characterization of Sclerotium rolfsii Causing Seedling Blight in Rice

In Telangana state, India, around June 2021, the seedling blight of rice was seen in nursery beds 10 days after sowing (DAS). Sclerotium rolfsii was the disease's causative agent. Infected plant samples were gathered in farmer's paddy nurseries in the Telangana state of India during kharif 2021 as part of the Rice Production Oriented Survey (POS) in farmer fields in Katkur (Village), Akkannapeta (Mandal), Siddipet (District). The farmer uses a seed called PAN 152 from a private company. The affected rice seedlings showed the typical signs of soft rot, yellowing and white cottony mould growth. Spherical-shaped dark brown sclerotial bodies were also seen in the basal stem region of the infected seedlings. Cultural and morphological identification was done for identifying the microbe.

First Report of Colletotrichum graminicola Causing Maize Anthracnose in Galicia, Northwestern Spain.

Maize (Zea mays) is one of the most important crops worldwide, and fungal diseases are responsible for major losses in food production. Anthracnose caused by Colletotrichum graminicola can infect all maize tissues, although stalk rot and seedling blight cause more significant economic damage (Munkvold and White, 2016). Anthracnose stalk rot is characterized by a distinctive external blackening of the lower stalks resulting in large black streaks, while the pith turns dark brown and has a shredded appearance. Like most stalk rots, the most obvious symptom is a sudden death of plants before grain maturity, and plant lodging. Symptoms commonly appear late in the season, suspicious maize stems of cultivar Tuy exhibiting symptoms of anthracnose stalk rot were collected from a field in Pontevedra, Galicia, Spain (Geographical coordinates: 42°23'27.1" N - 8°30'46.3" W) between June and December of 2022. Stem samples, approximately 50 mm2, were dissected and surface-disinfected for 90 seconds in 20% sodium hypochlorite (v/v) and rinsed three times in sterile distilled water. The samples were transferred to one half-strength acidified potato dextrose agar (PDA) supplemented with ampicillin (100 µg/mL) and lactic acid 90% (1.5 mL/L) and incubated for 5 days at 25 ºC (Sukno et al. 2008). Single spores were transferred to fresh PDA plates to obtain pure culture isolates. A total of six isolates were obtained, and among them, two were selected for further characterization (SP-36820-1 and SP-36820-3). Colonies grown on PDA have dark gray aerial mycelium with orange-colored spore masses. Conidia are falcate, slightly curved, tapered toward the tips, and are produced in acervuli with setae, measuring 37.65 to 24.84 x 8.02 to 4.67 µm, respectively (n = 100). These morphological characteristics are in agreement with C. graminicola previously described by Bergstrom and Nicholson (1999). Isolates were grown in potato dextrose broth (PDB) for 3 days at 25 ºC and total genomic DNA was extracted using a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA, USA). The internal transcribed spacer region of rDNA and the manganese-type superoxide dismutase gene (SOD2) were amplified using primers ITS4/ITS5 (White et al. 1990) and SOD625/SOD507 (Fang et al. 2002) and consequently sequenced. GenBank BLAST analysis revealed that the sequences were 100% identical to strains of C. graminicola. All sequences were deposited in GenBank (see e-Xtra 1 for accession numbers). To confirm Koch's postulates, plants of a derivative of maize inbred line Mo940 (developmental stage V3) were placed horizontally in a tray for inoculation and 20 droplets (7.5 µL) of a suspension of 3 x 105 conidia per milliliter were placed on the surface of the third leaf. The trays were closed to retain moisture and incubated overnight at 23ºC. The next day, the plants were returned to a vertical position and incubated in a growth chamber at 25ºC with 80% humidity and a light cycle of 16 h of light and 8 h of dark (Vargas et al. 2012). After four days inoculated leaves presented brown elongated lesions with necrotic centers consistent with C. graminicola infection, whereas control plants remained asymptomatic. The strains reisolated from infected leaves were morphologically identical to the original isolates. To our knowledge, this is the first report of Colletotrichum graminicola causing maize anthracnose in Spain. Recently, maize anthracnose was also reported in Bosnia and Herzegovina and China (Duan et al. 2019; Cuevas-Fernández et al. 2019), suggesting the pathogen's geographic range is increasing, which may be a threat to maize cultivation in locations with optimal humid conditions for disease development.

First report of seedling blight caused by Fusarium tricinctum on Cynanchum stauntonii in China.

Cynanchum stauntonii is a perennial herb plant of the Asclepiadaceae family. The dried roots and rhizomes have been used as medicine in China for 1500 years and are considered a remedy for cough and phlegm. In recent years, the wild C. stauntonii resources have not been sufficient for market demand, therefore, a large artificial cultivation area was established in Xinzhou, Tuanfeng and Macheng in Hubei province. In March and April 2022, serious outbreaks of seedling blight were observed in C. stauntonii in Xinzhou county (N30°48'12″, E114°49'24″), and the disease occurred on 10 to 15% of plants in five C. stauntonii nursery beds. Early symptoms included withered tips, chlorosis, stunting, yellow leaves and leaf drop, and later, seedlings die in patches. To determine the causal agent of disease, pieces (5 mm × 5 mm) of diseased tissue at the junction of disease and healthy tissue were surface disinfected by soaking in 75% ethanol for 3 min, rinsed three times with sterilized water, and pieces were placed on PDA at 25°C. Fungal isolates obtained were yellow-brown at the center and pink to white toward the periphery, and dark red pigments were observed in the agar. Isolates were cultured in synthetic low nutrient agar (SNA) and carnation leaf agar to observe the spore morphology. The macroconidia were sickle-shaped with 3-4 septate, with sizes of 30.26±2.36×3.77±0.53 μm on SNA and 33.52±2.20×3.81±0.48 μm on carnation leaf agar (n=30). Morphological characteristics of the isolates were consistent with those of Fusarium sp in the Fusarium Laboratory Manual (Leslie et al. 2006). Furthermore, the genomic DNA from a representative isolate BQ-2 was extracted, the ITS, TEF-1α, RPB1 and RPB2 genes were amplified with primers ITS1/ITS4, EF1/EF2, Fa/G2R and 5F2/7cr, respectively (Zhang et al. 2022). BLAST analysis showed that the ITS (ON935780.1), TEF-1α (OP985126.1), RPB1 (OP985125.1) and RPB2 (OP985124.1) amplicon sequence were 99.44%, 98.94%, 99.88% and 100% identical to the sequences of F. tricinctum strain (KU350724.1, AB674264.1, LC701712.1, MW474678.1), respectively. A phylogenetic tree constructed based on a concatenated sequence (ITS, TEF-1α, RPB1, RPB2) using the neighbor-joining and maximum likelihood method in MEGA7 revealed that BQ-2 grouped with concatenated sequences from four representative F. tricinctum isolates in GenBank. Based on the morphological characteristics and molecular identification, the strain BQ-2 was identified as F. tricinctum. For pathogenicity tests, 5 mm pieces of a BQ-2 colony on PDA were placed on excised leaves of healthy C. stauntonii wounded with a needle (n=5) and kept at 25±2℃. Leaves treated PDA were used as a control (Li et al.2020). After three days inoculation, the mycelia proliferated and began to infect leaf tissues. Ten days later, large parts of the detached leaves were extensively infested with the pathogen and brown. For live plant inoculation, stem bases of five healthy seedlings were punctured with sterile needle and then inoculated with BQ-2 mycelia from PDA. Controls were treated with only PDA. The seedlings began wilting after three days and at five days showed typical disease symptoms, similar to those observed in the field. The controls were asymptomatic. The pathogen was reisolated from the diseased tissues, and the colonies and microscopic characteristics were similar to those of BQ-2. To the best of our knowledge, this is the first report of F. tricinctum causing seedling blight on C. stauntonii in China. This report will provide resources and reference for controlling the increased incidence and economic losses of seedling blight on C. stauntonii.

Biocontrol Efficacy of Endophyte Pseudomonas poae to Alleviate Fusarium Seedling Blight by Refining the Morpho-Physiological Attributes of Wheat.

Some endophyte bacteria can improve plant growth and suppress plant diseases. However, little is known about the potential of endophytes bacteria to promote wheat growth and suppress the Fusarium seedling blight pathogen Fusarium graminearum. This study was conducted to isolate and identify endophytic bacteria and evaluate their efficacy for the plant growth promotion and disease suppression of Fusarium seedling blight (FSB) in wheat. The Pseudomonas poae strain CO showed strong antifungal activity in vitro and under greenhouse conditions against F. graminearum strain PH-1. The cell-free supernatants (CFSs) of P. poae strain CO were able to inhibit the mycelium growth, the number of colonies forming, spore germination, germ tube length, and the mycotoxin production of FSB with an inhibition rate of 87.00, 62.25, 51.33, 69.29, and 71.08%, respectively, with the highest concentration of CFSs. The results indicated that P. poae exhibited multifarious antifungal properties, such as the production of hydrolytic enzymes, siderophores, and lipopeptides. In addition, compared to untreated seeds, wheat plants treated with the strain showed significant growth rates, where root and shoot length increased by about 33% and the weight of fresh roots, fresh shoots, dry roots, and dry shoots by 50%. In addition, the strain produced high levels of indole-3-acetic acid, phosphate solubilization, and nitrogen fixation. Finally, the strain demonstrated strong antagonistic properties as well as a variety of plant growth-promoting properties. Thus, this result suggest that this strain could be used as an alternate to synthetic chemicals, which can serve as an effective method of protecting wheat from fungal infection.

Assessment of Tunisian Trichoderma Isolates on Wheat Seed Germination, Seedling Growth and Fusarium Seedling Blight Suppression.

Beneficial microorganisms, including members of the Trichoderma genus, are known for their ability to promote plant growth and disease resistance, as well as being alternatives to synthetic inputs in agriculture. In this study, 111 Trichoderma strains were isolated from the rhizospheric soil of Florence Aurore, an ancient wheat variety that was cultivated in an organic farming system in Tunisia. A preliminary ITS analysis allowed us to cluster these 111 isolates into three main groups, T. harzianum (74 isolates), T. lixii (16 isolates) and T. sp. (21 isolates), represented by six different species. Their multi-locus analysis (tef1, translation elongation factor 1; rpb2, RNA polymerase B) identified three T. afroharzianum, one T. lixii, one T. atrobrunneum and one T. lentinulae species. These six new strains were selected to determine their suitability as plant growth promoters (PGP) and biocontrol agents (BCA) against Fusarium seedling blight disease (FSB) in wheat caused by Fusarium culmorum. All of the strains exhibited PGP abilities correlated to ammonia and indole-like compound production. In terms of biocontrol activity, all of the strains inhibited the development of F. culmorum in vitro, which is linked to the production of lytic enzymes, as well as diffusible and volatile organic compounds. An in planta assay was carried out on the seeds of a Tunisian modern wheat variety (Khiar) by coating them with Trichoderma. A significant increase in biomass was observed, which is associated with increased chlorophyll and nitrogen. An FSB bioprotective effect was confirmed for all strains (with Th01 being the most effective) by suppressing morbid symptoms in germinated seeds and seedlings, as well as by limiting F. culmorum aggressiveness on overall plant growth. Plant transcriptome analysis revealed that the isolates triggered several SA- and JA-dependent defense-encoding genes involved in F. culmorum resistance in the roots and leaves of three-week-old seedlings. This finding makes these strains very promising in promoting growth and controlling FSB disease in modern wheat varieties.

First report of seedling stem blight of mango caused by Neofusicoccum parvum in Italy.

Seedling blight of mango (Mangifera indica L.) was observed in a commercial nursery located in Messina province (eastern Sicily, Italy) during winter of 2021. More than 30% of 3,000 seedlings, about three to six months old, of mango cv. Gomera 3 showed symptoms of basal stem blight. The symptoms started from seed, led to the decline and subsequent death of the plants. Necrotic lesions appeared at crown level two months after sowing. The stem tissues of ten symptomatic plants were cut, surface sterilized, dipped in 1.5% sodium hypochlorite for 1 min and transferred onto potato dextrose agar medium (PDA) and incubated at 25°C for four days. Approximately 60% of stem tissues developed very similar fungal colonies, resembling to Botryosphaeriaceae. A total of four representative isolates were collected through single hyphal-tip and stored at 4 °C. The internal transcriber spacer region (ITS) was amplified with primers, ITS5/ITS4 (White et al., 1990), and EF1-728F and EF1-986R (Carbone and Kohn, 1999) were used to amplify part of the translation elongation factor 1alpha gene (tef1-α), and primers Bt2a/Bt2b (Glass and Donaldson, 1995) were used for the partial β-tubulin (tub2). The obtained sequences were deposited in GenBank with accession numbers: ON911292-95 for the ITS, ON933621-24 for tef1-α and ON933625-28 for tub2.To compare the results, 50 additional sequences were selected and inserted in the alignment according to the recent literature on the Botryosphaeriaceae (Bezerra et al., 2021; Zhang et al., 2021). Maximum parsimony analysis (MP) of concatenated dataset (ITS + tef1-α + tub2) was performed in PAUP v.4.0a. Clade support was assessed by 1,000 bootstrap replicates and Botryosphaeria dothidea was used as an outgroup. Our isolates clustered within the group of Neofusicoccum parvum (71% bootstrap value) (ex-type CMW9081). Based on these results, and morphological data (50 conidia length × width average: 18.1 × 6.6, respectively) our isolates (named MC) were identified and confirmed as Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. Pathogenicity tests were also conducted on 18 mango cv. Gomera 3 seedlings. The crown roots of each seedling were mechanically wounded and a mycelial plug of the isolate MC14 was placed onto them and covered with soil. Controls (three seedlings) were inoculated with sterile PDA only. Seedlings were maintained in a growth chamber with a 12 hrs photoperiod at 25°C ± 1°C and watered regularly. After five days, stem lesions appeared externally (1.6 cm) and one month after the inoculation, all the inoculated seedlings died. However, controls did not show any obvious symptoms. Re-isolations were conducted as described above and fulfilled Koch's postulates confirming pathogenicity. Among the diseases affecting mango plants, Botryosphaeriaceae represent a serious threat in Sicily as reported by Aiello et al., 2022. The endophytic behaviour of Botryosphaeriaceae is well known, making them latent pathogens (Slippers and Wingfield, 2007). In Italy, N. parvum was detected in mango orchards since 2013 (Ismail et al. 2013), but symptoms of seedlings stem blight have never been reported in the nursery. In Sicily, an increase of Botryosphaeriaceae infection has been observed recently, especially in nurseries, where N. parvum has been identified as a most destructive pathogen (Aiello et al., 2020; Gusella et al., 2021). To our knowledge, this is the first report worldwide of N. parvum causing mango seedling blight. The high incidence of infected seedlings detected in this study highlights the potential risk during propagation in the nursery, representing a significant source of inoculum for the field.

DETECTION OF Acidovorax citrulli IN WATERMELON SEEDLINGS IN HOPELCHÉN, CAMPECHE, MEXICO

In Mexico, there are no reports of the presence of Acidovorax citrulli, the causal agent of bacterial fruit blotch and seedling blight in cucurbits. The introduction of A. citrulli is a latent risk due to the importation of seeds and seedlings. Fruits and plants with typical symptoms of this disease have been observed in watermelon (Citrullus lanatus) production fields of small growers and commercial nurseries in the municipality of Hopelchén, Campeche. The objective was to detect A. citrulli by PCR in watermelon seedlings in a production area in Hopelchén, with the hypothesis that the bacterium is present in this municipality. In December 2018, 51 samples of nine watermelon varieties were collected from two seedling production nurseries. Each sample contained ten symptomatic seedlings ranging in age from 12 to 41 days. End-point PCR analysis with primers WFB1 and WFB2 indicated the presence of A. citrulli in 68.6 % of the samples; immuno-PCR results confirmed the presence of the bacterium in 80.9 % of these samples. Consensus 16S rRNA gene fragment sequences from eight samples aligned with 98 % similarity on average to A. citrulli sequences deposited in the GenBank-NCBI database. The dark, sunken, elongated lesions on cotyledonal leaves, as well as the black, spreading lesions from the margin to the base of true leaves observed on watermelon seedlings, were caused by A. citrulli. This study constitutes a new report of A. citrulli in Campeche, which should be considered to delimit and prevent the spread of the bacterium to other cucurbit-producing areas in Mexico.

Emergence and Morphological Response of Cashew (Anacardium occidentale L.) Treated Nuts as Influenced by Some Fungicides on Young Seedlings

Cashew (Anacardium occidentale L.) seedlings are attacked by fungi diseases such as damping off and seedling blight caused by fungi such as Fusarium spp. and Rhizoctonia spp. which can amount to about 60-65% loss in the nursery. Cashew nut seeds are majorly sown by farmers untreated. Fungicides have also been observed to delay seedling emergence and negatively influence morphological traits in some crops. This experiment aims at observing the effect of using fungicide seed dressings on cashew seedling emergence and morphology before transplanting. Medium cashew nut biotype and three commonly used fungicides were used. The 3 months experiment was set up in the nursery using a Completely Randomized Design CRD. The treatments are; Control + Medium (Ct); Apron Star + Medium (AS), Dress Force + Medium (DF) and Seed Care + Medium (SC). Topsoil filled perforated nursery polythene bags were used. No significant difference was identified in all morphological traits observed in the experiment among the treatments all through the duration of the experiment. From a maximum vigour scale of 5, Ct, AS and DF all had a seedling vigour of 4.7 while SC had 4.2. As a precautionary measure against fungi diseases, Apron Star, Dress Force and Seed Care have been observed not to have a negative effect on the emergence and growth of young cashew seedlings when nuts are treated with them before sowing.

Aggressiveness and mycotoxin profile of Fusarium avenaceum isolates causing Fusarium seedling blight and Fusarium head blight in UK malting barley.

Fusarium avenaceum causing Fusarium seedling blight (FSB) and Fusarium head blight (FHB) on barley is associated with economic losses of crop yield and quality, and the accumulation of mycotoxins including the enniatins (ENNs) A, A1, B and B1. Although F. avenaceum is the main producer of ENNs, studies on the ability of isolates to cause severe Fusarium diseases or produce mycotoxins in barley are limited. In this work, we investigated the aggressiveness of nine isolates of F. avenaceum to two cultivars of malting barley, Moonshine and Quench, and defined their ENN mycotoxin profiles in in vitro and in planta experiments. We assessed and compared the severity of FSB and FHB caused by these isolates to disease severity by F. graminearum, F. tricinctum and F. poae. Quantitative real-time polymerase chain reaction and Liquid Chromatography Tandem Mass Spectrometry assays were used to quantify pathogen DNA and mycotoxin accumulation, respectively, in barley heads. Isolates of F. avenaceum were equally aggressive to barley stems and heads and caused the most severe FSB symptoms resulting in up to 55% reductions of stem and root length. Fusarium graminearum caused the most severe FHB disease, followed by the isolates of F. avenaceum with the most aggressive F. avenaceum isolates capable of causing similar bleaching of barley heads as F. avenaceum. Fusarium avenaceum isolates produced ENN B as the predominant mycotoxin, followed by ENN B1 and A1 in vitro. However, only the most aggressive isolates produced ENN A1 in planta and none produced ENN A or beauvericin (BEA) either in planta or in vitro. The capacity of F. avenaceum isolates to produce ENNs was related to the accumulation of pathogen DNA in barley heads, whilst FHB severity was related to the synthesis and accumulation of ENN A1 in planta. Cv. Moonshine was significantly more resistant than Quench to FSB or FHB, caused by any Fusarium isolate, and to the accumulation of pathogen DNA, ENNs or BEA. In conclusion, aggressive F. avenaceum isolates are potent ENN producers causing severe FSB and FHB with ENN A1 requiring further investigation as potential virulence factor for F. avenaceum in cereals.