First Report of Fusarium meridionale Causing Stalk Rot of Ryegrass in China

Members of the Fusarium graminearum species complex (FGSC) are the main causing agents of head blight, seedling blight, or stalk rot in wheat and other cereals worldwide. Surveys on species composition and mycotoxin production of FGSC populations have mainly focused on food crops such as wheat, maize, and barley, but little is known about the identity of FGSC pathogens present in pasture grass. In April 2021, a survey of grass diseases in the Hongya County (29.90661 N; 103.37313 E) in Sichuan Province was conducted to understand the etiology of stalk rot in perennial ryegrass (Lolium perenne). It was observed in several pastures that about 10% of yield loss in perennial ryegrass was caused by stalk rot. Affected plant stalks were brown to dark brown in colour and appeared soggy. As infections continued or under conditions of high humidity, some plant stalks also became flattened. Perennial ryegrass samples with symptoms of stalk rot or browning of the stem were collected. Symptomatic tissues were cut into short segments (approximately 5 mm), surface-sterilized in 3% sodium hypochlorite solution for 2 min, rinsed three times with sterile distilled water, air dried, plated onto potato dextrose agar (PDA), and then incubated in the dark at 28 °C. After 3 to 5 days, Fusarium-like fungal colonies with reddish-orange mycelium were collected and transferred to new PDA plates for further purification, and the purified cultures were obtained by single spore isolation. Four uniform isolates were obtained and their colonies on PDA resembled typical FGSC colonies (Leslie and Summerell 2006; O’Donnell et al. 2004). Colonies had an average radial growth rate of 8.5 to 11.0 mm/day at 28 °C in the dark on PDA. Conidial characteristics were studied on Spezieller Nährstoffarmer agar (SNA) as described by Wang et al. (2014). Macroconidia were falcate to almost straight, usually with parallel dorsal and ventral lines, 3- to 5-septate, 20.65 to 55.22 μm in length (average 39.16 μm), and 2.38 to 6.93 μm in width (average 4.42 μm) (n = 200). No microconidia were observed. The pathogenicity of the isolated Fusarium strains was then tested on healthy perennial ryegrass (variety Changjiang 1). Ryegrass plants grown for 2 months were inoculated by punching a hole in the stem using a sterile toothpick, followed by an injection of 20 μL macroconidia suspension at a concentration of 105 spores/mL. Ryegrass stems treated with water served as the control. Twenty plants were included in each treatment. After inoculation, the plants were grown in a growth chamber at 25 °C and 90% humidity for 24 h. Stalk tissues at the wound site turned brown after 3 days and the brown area then extended to regions above and below. No symptoms were observed in the water-treated controls. As well, the same pathogen was reisolated from the infected grass stems, but not from the controls. Thus, the isolated Fusarium spp. are a cause of stalk rot in perennial ryegrass based on the fulfillment of Koch’s postulates. To identify the Fusarium spp. to species level, portions of the translation elongation factor 1-α (TEF) gene sequences from all four strains were amplified and sequenced as described by Wang et al. (2015). The obtained sequences were identical, and a sequence of isolate SC1 was submitted to GenBank (accession no. MZ964308). BLASTn searches were conducted on the TEF sequence (607 bp) in two databases, revealing it had 100% similarity to the sequence of Fusarium meridionale strain DS27 (accession no. MN629330) in NCBI and strain NRRL28723 from FUSARIUM-ID (http://isolate.fusariumdb.org/). A concatenated four-gene phylogeny (supplementary figure) resolved SC1 and the type specimens of F. meridionale (NRRL28723, 29010, and 28436) in a monophyletic clade with 100% bootstrap support, confirming that the strain SC1 belongs to F. meridionale. Finally, trichothecene productions of F. meridionale strains were evaluated using rice cultures kept at 28 °C in the dark for two weeks, as described by Desjardins and Proctor (2011). LC-MS/MS analysis indicated that the fungus could produce NIV and 4ANIV in rice cultures with average concentrations of 1400.44 and 3144.10 μg/kg, respectively. To the best of our knowledge, this is the first report of F. meridionale causing disease in perennial ryegrass in China. Further research will be necessary to determine its distribution, aggressiveness, and trichothecene production.

Sorghum sheath blight caused by Fusarium spp. in Sinaloa, Mexico

Sorghum (Sorghum bicolor) leaf sheath blight was observed for the first time in Sinaloa, Mexico in the summer of 2020. Fungal isolates were obtained from symptomatic tissue in PDA. Fusarium spp. were associated with symptomatic plants in ten sampling sites under field conditions. No root and stalk rot were observed during the sampling period. Analysis of fragments of the EF-1a and RPB2 genes indicated that all isolates belong to the Fusarium fujikuroi species complex (FFSC). Five groups were delineated from this complex: F. thapsinum, F. verticillioides, Fusarium sp. (four isolates), Fusarium sp. (Fus4), and Fusarium sp. (Fus16), which is closely related to Fusarium madaense. The morphological characteristics (colony color and morphometry of conidia) of isolates with sequence similarities to those of F. thapsinum and F. verticillioides were in the expected range for these species. The morphology of isolates Fus7a, Fus7b, Fus11 and Fus17, as well as Fus4 and Fus16, were similar to those of the FFSC, specially to F. andiyazi and F. madaense. Inoculations of sorghum with representative isolates of F. thapsinum, F. verticillioides and the unidentified Fusarium species resulted in reddish brown lesions similar to those observed under field conditions; the original isolates inoculated were reisolated fulfilling the Koch's postulates. Although leaf sheaths on sorghum plants were heavily damaged, root and stalk rot were not observed in the greenhouse inoculations or under field conditions. Future research should focus on determining the identity of the unknown Fusarium spp. in order to design control measures of the disease. This is the first report of Fusarium spp. causing sorghum leaf sheath blight in Mexico.

Multiple Species of Asteraceae Plants are Susceptible to Root Infection by the Necrotrophic Fungal Pathogen Sclerotinia sclerotiorum

The necrotrophic fungal pathogen Sclerotinia sclerotiorum can cause disease on numerous plant species, including many important crops. Most S. sclerotiorum-incited diseases of crop plants are initiated by airborne ascospores produced when fungal sclerotia germinate to form spore-bearing apothecia. However, basal stalk rot of sunflower occurs when S. sclerotiorum sclerotia germinate to form mycelia within the soil which subsequently invade sunflower roots. To determine if other plant species in the Asteraceae family are susceptible to root infection by S. sclerotiorum, cultivated sunflower (Helianthus annuus L.) and seven other Asteraceae species were evaluated for S. sclerotiorum root infection by inoculation with either sclerotia or mycelial inoculum. Additionally, root susceptibility of sunflower was compared to that of dry edible bean and canola, two plant species susceptible to S. sclerotiorum but not known to display root-initiated infections. Results indicated that multiple Asteraceae family plants are susceptible to S. sclerotiorum root infection after inoculation with either sclerotia or mycelium. These observations expand the range of plant hosts susceptible to S. sclerotiorum root infection, elucidate differences in root inoculation methodology, and emphasize the importance of soil-borne infection to Asteraceae crop and weed species.

The Impact of Climate Change on Changing Patten of Maize Diseases in Indian Subcontinent: A Review

Climate change influences the occurrence, prevalence, and severity of plant pathogens. Global temperatures are predicted to rise by 2–4°C due to human activities and increased market globalization, coupled with rising temperatures, leads to a situation favorable to pest movement and establishment. Maize is an important crop after wheat and rice. Changes in rainfall distribution and temperature may result in temporary excessive soil moisture or water logging or drought in some maize producing areas leading to alterations in biotic stress factors. In Indian subcontinent warming trend in climate along the west coast, central, interior peninsula and northeast regions creates favorable conditions for diseases in maize like sorghum downy mildew (SDM) and Turcicum leaf blight (TLB). The decreasing trend of monsoon, seasonal rainfall in North India, Central India, parts of Gujarat and Kerala is suitable for post flowering stalk-rot (PFSR) which is gaining importance in maize. The outcome for any host-pathogen interaction under changing climate is not readily predictable. This review assesses the potential effects of climate change on maize pathogens and consequently on plant health. The evidence assessed indicates that climate change has already expanded pathogen’s host range and geographical distribution increasing the risk of introduction of pathogens into new areas.

Genetic Testing of Inbred Lines and Single Cross Hybrids against Fusarium Stalk Rot Caused by Fusarium moniliforme in Maize (Zea mays L.)

Globally, Maize (Zea mays L.) is a third major cereal food crop. It is a multipurpose crop with 26% of its production is used as food by human beings. Maize is known as “queen of cereals”, because of its high genetic yield potential, efficient utilization of radiant energy and wider adaptability. About 65 different phytopathogens affect the maize production in different stages of life cycle. Among which Fusarium moniliforme is one such soil borne pathogen causes Fusarium stalk rot (FSR) disease that ultimately reduces maize yield potential over the world. In any breeding program, screening and genetic testing of available germplasm resources against pathogens is necessary to prevent yield losses. Hence, the present research screened around 114 maize inbred lines and 45 single cross hybrids (SCHs) against FSR under artificial epiphytotic conditions. Among 114 inbreds, only four inbreds viz., CM 202, 10878, MAI-759 and MAI-766 (mean disease score of 3-4) showed moderately resistant reaction and out of 45 SCHs, only one hybrid combination i.e., MAI329 × CM202 (mean disease score was 2.60) exhibited resistance reaction against Fusarium stalk rot. Nevertheless, these resistance sources could be utilized in maize breeding programs for obtaining high yielding cultivars with resistance towards FSR disease.

Physiological Influence of Stalk Rot on Maize Lodging after Physiological Maturity

The stalk lodging caused by stalk rot after physiological maturity (PM) is a major factor restricting further development of mechanical grain harvesting in China. The physiological mechanism of stalk rot on maize stalk lodging after PM is not clear. This study, based on investigating stalk rot under natural field conditions, demonstrated the relation between stalk rot caused by Fusarium spp. and lodging of 35 maize cultivars after PM. In addition, three widely-planted maize cultivars were inoculated with Fusarium spp. at PM to analyze the pathogen of stalk rot causing lodging, by measuring the infection process, carbohydrate contents, and mechanical strength of stalks. Stalk lodging increased by 0.11–0.32% for each 1% incidence of stalk rot. The stalk rot pathogen infected stalks from the pith to the rind. At the level of longitudinal section, the stalk rot pathogen spread from the inoculation internode upwardly and downwardly. These infections gradually increased with the days after PM. Inoculated plants had decreased soluble sugar content; however, cellulose and lignin contained in the inoculated plants were both higher than that in the non-inoculated treatment. Crushing strength was significantly and positively correlated with percentage of soluble sugar. This indicated that the reduction of soluble sugar content during the natural senescence of maize stalk after PM was an important factor for the decrease of stalk strength and the increase of stalk lodging. The occurrence of stalk rot accelerated the decomposition of soluble sugar, which accelerated the decrease of stalk strength and greatly increased risk of stalk lodging.

The presence of bacterial stalk rot disease on corn in Indonesia: A review

Bacterial stalk rot disease in corn results in a significant reduction in yield due to the interruption of the flow of nutrients from the roots to other parts of the plant. Pathogenic bacteria infect the inner tissue of the stalk until it rots. This disease has been reported to attack corn crops in Asia and Europe such as India, Korea, Thailand, Philippines, Nepal, Mexico, Serbia, and China. In Indonesia, this disease was first reported to attack corn in the West Sulawesi region by the Mamuju Class II Quarantine Station. The results of molecular identification indicated that this disease is caused by the bacterium Dickeya zeae, previously known as Erwinia chrysanthemi pv. zeae that previously reported attacked pineapple and aloe vera in Indonesia. The potential for economic losses due to this disease is quite high, so appropriate and efficient control measures are needed. Based on those, this research study about the symptom, the characteristic of the bacteria agent caused the stalk rot disease, the distribution and the impact to the maize production in Indonesia.

First Report of Fusarium culmorum Causing Maize Stalk Rot in China

Maize stalk rot has become one of the most important diseases in maize production in China. From 2017 to 2019, a survey was conducted to determine the population diversity of Fusarium species associated with maize diseases in 18 cities across Henan Province. Maize stalk rot with an incidence of more than 20% that caused yield losses up to 30% was observed on maize variety Zhengdan958, which was grown in two continuous maize fields in Zhumadian City, Henan Province. The stem tissues from the boundary between diseased and healthy pith were chopped into small pieces (3 × 8 mm), disinfected (70% ethanol for 1 min) and then placed onto potato dextrose agar (PDA) amended with L-(+)-Lactic-acid (1 g/L) and incubated at 25°C for 4 days. Colonies on PDA produced fluffy, light yellow aerial mycelium and purple to deep brick red pigment at 25°C (Fig 1A, 1B). On carnation leaf agar (CLA), macroconidia in orange sporodochia formed abundantly, but microconidia were absent. Macroconidia were short and thick-walled, had 3 to 5 septa, a poorly developed foot cell and rounded apical cell (Fig 1C). These characteristics matched the description of Fusarium culmorum (Leslie and Summerell 2006) and isolates DMA268-1-2 and HNZMD-12-7 were selected for further identity confirmation. Species identification was confirmed by partial sequences of three phylogenic loci (EF1-α, RPB1, and RPB2) using the primer pairs EF1/EF2, CULR1F/CULR1R, and CULR2F/CULR2R, respectively (O'Donnell et al., 1998). The consensus sequences from the two isolates were deposited in GenBank (MZ265416 and MZ265417 for TEF, respectively; MZ265412 and MZ265414 for RPB1, respectively; MZ265413 and MZ265415 for RPB2). BLASTn searches indicated that the nucleotide sequences of the three loci of the two isolates revealed 99% to 100% similarity to those of F. culmorum strains deposited in the GenBank, Fusarium-ID, and MLST databases (Supplementary Table 1~3). Pathogenicity test was conducted at the flowering-stage using Zhengdan958 and Xundan20 plants according to previously described method (Zhang et al., 2016; Cao et al., 2021; Zhang et al., 2021). The second or third internodes of thirty flowering plants were drilled to make a wound approximately 8 mm in diameter using an electric drill. Approximately 0.5 mL inoculum (125 mL colonized PDA homogenized with 75 mL sterilized distilled water) was injected into the wound and sealed with Vaseline and Parafilm to maintain moisture and avoid contamination. Sterile PDA slurry was used as a control. Thirty days after inoculation, the dark-brown, soft rot of pith tissues above and below the injection sites were observed, and some plants were severely rotten and lodged (Fig 1D, 1E). These symptoms were similar to those observed in the field. No symptoms were observed on control plants. The same pathogen was re-isolated from the inoculated stalk lesions but not from the control, thereby fulfilling Koch's postulates. To our knowledge, this is the first report of F. culmorum as the causal agent of stalk rot on maize plants in China. Also, this fungus has been reported to cause maize ear rot in China (Duan et al. 2016) and produce mycotoxins such as trichothecenes, nivalenol, and zearalenone that cause toxicosis in animals (Leslie and Summerell 2006). The occurrence of maize stalk rot and ear rot caused by F. culmorum should be monitored due to the potential risk for crop loss and mycotoxin contamination.

First Report of Stalk Rot of Celery Caused by Erwinia rhapontici in China

Species belonging to the genus Erwinia cause diseases in many economically important plants (Mansfield et al. 2012). In May 2021, celery plants (Apium graveolens var. dulce) showing soft rot symptoms were observed in greenhouses (cv. Queen of France) in Boye County, Baoding, Hebei Province (North China). Disease symptoms began with pinkish water-soaked lesions on the midrib of celery stalks, but at the same time the leaves and root did not show symptoms. The infected celery plants rapidly developed brownish rotten stalks and leaves turned dry and yellow, but root remained asymptomatic. The disease incidence in two greenhouses (0.15 ha in size) was more than 50%. Affected celery stalk tissues were cut into 0.5 cm pieces, followed by surface sterilization using 75% ethanol for 60 sec and then three successive rinses with sterile distilled water. Then, the tissues were immersed in 200 µl 0.9% saline for 15 min. Aliquots of two tenfold dilutions of the tissue specimen soaking solution were plated onto Luria-Bertani (LB) agar plates and incubated at 28°C for 24 h. Single colonies were picked and restreaked onto LB agar three times for purity. The bacterial gDNA was extracted using the EasyPure Bacteria Genomic DNA Kit (TransGen Biotech). The 16S rDNA region was amplified by PCR using the universal primers 27F/1492R and sequenced. Result of blastn analysis of the 16S rDNA amplicons (MZ489246-MZ489247) indicated that the bacterial isolates (BY21311 and BY21312) belonged to the genus Erwinia. Biolog analysis (GEN III Microplate) identified the two isolates BY21311 (SIM=0.668) and BY21312 (SIM=0.638) as E. rhapontici. Housekeeping genes including acnA, gapA, icdA, mdh and rpoS were also amplified using a set of PCR primers (Ma et al. 2007; Waleron et al. 2008) followed by sequencing (MZ463029-MZ463038). To determine the species of the Erwinia isolates BY21311 and BY21312, multi-locus sequence analysis (MLSA) was performed with five housekeeping genes, and phylogenetic tree was reconstructed using RAxML v8.2.12 (Stamatakis et al. 2005). No sequence variation was observed at any MLSA locus between BY21311 and BY21312. The result of phylogenetic analysis showed that the celery stalk rot isolates BY21311 and BY21312 were clustered with E. rhapontici isolates. These celery isolates are closely related to the cabbage (Brassica rapa) isolate MAFF311153 (AP024329.1) in Japan. When celery plants have eight to nine true leaves, plants (cv. Queen of France) were inoculated with the isolate BY21311 by injecting 20 µl of bacterial suspensions (106 CFU·mL-1) into the celery stalks, or injected with 20 µl of 0.9% saline as control. The seedlings were grown at 25 °C and 50% relative humidity. Three days after inoculation, only infected seedling showed disease symptoms resembled to those observed in greenhouses. Bacterial colonies were obtained from the infected stalks and were identified using the same PCR primers of housekeeping genes as described above, fulfill Koch’s postulates. E. rhapontici has been reported to cause pink seed, crown and stem rot, soft rot or leaf spot on many plant hosts including pea (Pisum sativum), chickpea (Cicer arietinum), lentil (Lens culinaris), common bean (Phaseolus vulgaris), lucerne (Medicago sativa), wheat (Triticum aestivum), hyacinth (Hyacinthus orientalis), onion (Allium cepa), kiwifruit (Actinidia chinensis) and peach (Prunus persica) (Huang et al. 2003; Wang et al. 2017; Zhang et al. 2018; Kovács et al. 2020). To our knowledge, this is the first report of E. rhapontici causing stalk rot in celery. Stalk rot of celery has increased in prevalence over recent years in the Baoding region, it can cause significant yield loss and no cultivar has been found to be resistant to this disease so far. The stalk rot poses significant threat to local celery production, and further research on epidemiology and disease management options is needed.