Bulb Rot Research Articles

The Influence of Sprinkler and Drip Irrigation on the Incidence and Severity of Bacterial Disease in Onions Grown in Northeast California

Overhead sprinkler irrigation and its effect on leaf wetness are often referenced as a key factor for bacterial disease spread in onions grown in arid climates. A study was conducted in 2021 and 2022 in Tulelake, California, to compare the incidence and severity of bacterial disease under sprinkler and drip irrigation. The experiment was a split-plot randomized complete block design with drip and solid-set sprinkler irrigation as the main plot treatments and inoculation and no inoculation with plant-pathogenic bacteria as the split-plot treatments. The entire study area was irrigated with solid-set sprinklers from planting until the five-leaf stage to facilitate uniform onion germination and chemigation of herbicides for weed control. After the five-leaf stage, irrigation treatments were imposed for the remainder of the growing season. Total irrigation water amount and irrigation frequency were similar for both irrigation methods. Foliar disease severity and the incidence of bulb rot were 170 and 186% higher, respectively, for solid-set sprinkler compared with drip irrigation. Leaf wetness within the plant canopy during onion bulbing was on average 36% higher in the solid-set sprinkler compared with drip irrigation treatment, whereas relative humidity and temperature were similar between irrigation methods. Drip irrigation maintained lower soil water tension and increased average bulb size by 9% compared with solid-set sprinkler irrigation. This study confirms historical observations that overhead irrigation can extend leaf wetness and increase bacterial disease in onions in arid production areas and highlights the potential benefits of the adoption of drip irrigation.

Biological Control of Fusarium oxysporum, the Causal Agent of Fusarium Basal Rot in Onion by Bacillus spp.

<i>Fusarium oxysporum</i> is the main pathogen causing Fusarium basal rot in onion (<i>Allium cepa</i> L.), which incurs significant yield losses before and after harvest. Among management strategies, biological control is an environmentally safe and sustainable alternative to chemical control. In this study, we isolated and screened bacteria for antifungal activity against the basal rot pathogen <i>F. oxysporum</i>. Isolates 23-045, 23-046, 23-052, 23-055, and 23-056 significantly inhibited <i>F. oxysporum</i> mycelial growth and conidial germination. Isolates 23-045, 23-046, 23-052, and 23-056 suppressed the development of Fusarium basal rot in both onion seedlings and bulbs in pot and spray inoculation assays. Isolate 23-055 was effective in onion seedlings but exhibited weak inhibitory effect on onion bulbs. Based on analyses of the 16S rRNA and <i>rpoB</i> gene sequences together with morphological analysis, isolates 23-045, 23-046, 23-052, and 23-055 were identified as <i>Bacillus thuringiensis</i>, and isolate 23-056 as <i>Bacillus toyonensis</i>. All five bacterial isolates exhibited cellulolytic, proteolytic, and phosphate-solubilizing activity, which may contribute to their antagonistic activity against onion basal rot disease. Taken together <i>B. thuringiensis</i> 23-045, 23-046, 23-052, and 23-055 and <i>B. toyonensis</i> 23-056 have potential for the biological control of Fusarium basal rot in onion.

Examination of Volatile Signatures of Fusarium Bulb Rot in Garlic Using Proton-Transfer-Reaction Time-of-Flight Mass Spectrometry and Solid-Phase Microextraction Gas Chromatography/Mass Spectrometry

Fusarium bulb rot, caused by Fusarium proliferatum, is a worldwide disease of garlic, both in the open field and during storage. Early diagnosis of the disease during storage is difficult due to the morphology of the bulbs and cloves. Volatile organic compounds (VOCs) are secondary metabolites produced by several microorganisms, including phytopathogenic fungi and bacteria. In recent years, the development of several techniques for the detection and characterization of VOCs has prompted their use, among others, as a diagnostic tool for the early and non-destructive analysis of many diseases of species of agricultural interest. In this paper, proton-transfer-reaction time-of-flight mass spectrometry (PTR-ToF-MS) and solid-phase microextraction gas chromatography/mass spectrometry (SPME-GC/MS) were successfully utilized to characterize the volatolome of commercial garlic cloves, artificially and naturally infected with F. proliferatum, for the early discrimination between diseased and healthy ones. A partial least squares discriminant analysis (PLSDA) and a principal component analysis (PCA) allowed for the separation of infected and healthy cloves and the identification of specific VOCs produced by the fungus during the infection. The results obtained in this work could be utilized for the development of simpler, more economical, and more portable devices for the early detection of infected garlic bulbs during storage.

First report of Anthracnose caused by Colletotrichum theobromicola on Cyclamen persicum in South Carolina, USA.

Florist's cyclamen (Cyclamen persicum) is an herbaceous perennial native to the Mediterranean region and has become an increasingly popular plant around the world. Leaves of these plants are cordate-shaped with varying green and silver patterns. Flowers vary in color from white through different shades of pink, lavender, and red. In September 2022, symptoms of anthracnose including leaf spots and chlorosis, wilting, dieback, and crown and bulb rot were observed on 20 to 30% of approximately 1,000 cyclamen plants in an ornamental production nursery in Sumter County, SC. Tissue samples surrounding the necrotic crowns were excised and sterilized in 10% bleach for 1 min, rinsed in sterile water, placed onto acidified potato dextrose agar (APDA), and incubated at 25°C with 24-h photoperiod. A total of five Colletotrichum isolates, 22-0729-A, 22-0729-B, 22-0729-C, 22-0729-D, and 22-0729-E were obtained by transferring hyphal tips to new plates. The morphology of these five isolates was identical, observed as gray and black with aerial gray-white mycelia and orange-colored spore masses. Conidia (n=50) measured 19.4 ± 5.1 mm (11.7 to 27.1 mm) in length and 5.1 ± 0.8 mm (3.7 to 7.9 mm) in width. Conidia were tapered with rounded ends. Setae and irregular appressoria were infrequently observed in aged cultures (> 60-day-old). These morphological features resembled those of members of the Colletotrichum gloeosporioides species complex (Rojas et al. 2010; Weir et al. 2012). Sequence of the internal transcript spacer (ITS) region of a representative isolate 22-0729-E (GenBank accession No. OQ413075) is 99.8% (532 / 533 nt) and 100% (533 / 533 nt) identical to those of the ex-neotype of Co. theobromicola CBS124945 (JX010294) and the ex-epitype of Co. fragariae (= Co. theobromicola) CBS 142.31 (JX010286), respectively. Its glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene sequence is 99.6% (272 / 273 nt) identical to those of CBS124945 (JX010006) and CBS 142.31 (JX010024). Its actin (ACT) gene sequence shares 99.7% (281 / 282 nt) and 100% (282 / 282 nt) identities with those of CBS124945 (JX009444) and CBS 142.31 (JX009516), respectively. Lastly, its beta-tubulin 2 (TUB2) gene sequence is 99.6% (704 / 707 nt) and 100% (707 / 707 nt) identical to those of CBS124945 (JX010447) and CBS 142.31 (JX010373), respectively. The causal agent causing anthracnose on cyclamen in SC was identified as Co. theobromicola. To confirm the pathogenicity, cyclamen 'Verano Red' plants grown in 2.5-inch pots were used in two pathogenicity assays using different inoculation methods. In the first assay, three plants were inoculated by spraying a conidial suspension (1 × 106 conidia per ml; 30 ml per plant) of isolate 22-0729-E onto the foliage. Three non-inoculated control plants were sprayed with distilled water. All six plants were placed in a plastic tray with wet paper towels. The tray was placed at 22°C for an 8-h photoperiod and covered for 7 days to maintain humidity. Early symptoms including small spots, marginal necrosis, and chlorosis were observed on leaves and flowers 8 days after inoculation (DAI) and the entire aboveground tissues of inoculated plants were blighted 13 to 21 DAI. Non-inoculated plants remained asymptomatic. In the second assay, sterile toothpicks were used to slightly wound the crown and bulb surface of three plants and secure a mycelial APDA plug of isolate 22-0729-E (5×5 mm2) onto each wound (three wounds per plant). Three control plants were wounded in the same manner, while sterile APDA plugs were used in place of mycelial plugs. All six plants were maintained in the same manner as in the first assay. Apparent leaf yellowing and wilting symptoms appeared as early as 13 DAI. On 21 to 28 DAI, severe crown rot on inoculated plants caused the entire foliage to collapse. At least one third of the inner crown and bulb tissues of each inoculated plant were rotten, while those of non-inoculated plants appeared healthy. Each assay was repeated once. Colletotrichum isolates resembling morphological characters of 22-0729-E were recovered from leaves and inner crown tissues of all inoculated plants in both assays, respectively, but not from non-inoculated control plants. Anthracnose diseases on Cyclamen persicum caused by Co. theobromicola (syn. Co. fragariae) have been reported in NC, USA (Lui et al. 2011) and Israel (Sharma et al. 2016). This is the first report of anthracnose on cyclamen in SC, USA. Colletotrichum gloeosporioides (teleomorph Glomerella cingulate) species complex on cyclamen has also been reported in Argentina (Wright et al. 2006), South Africa, and several other U.S. states (Farr and Rossman 2022). However, it remains unknown whether these previous reports in fact attributed to Co. theobromicola due to lack of molecular identification (Weir et al. 2012). Colletotrichum theobromicola can cause diseases on at least 30 other agricultural and horticultural crops such as strawberry, cacao, and boxwood (Farr and Rossman 2022). It may pose a threat to cyclamen in greenhouse and nursery productions. Therefore, management strategies are warranted in the future.

First report of Fusarium clavum causing garlic bulb rot in Mexico.

In Mexico, the main states for garlic (Allium sativum L.) production are Zacatecas, Guanajuato, and Puebla. In the 2020 crop season, garlic cultivation encompassed 6,794 ha, yielding 85,505 tons (SIAP, 2021). In February 2020, 35 garlic samples showing basal rot symptoms were collected from the garlic-growing regions in the states of Zacatecas and Aguascalientes in the municipalities of San Antonio Tepezala 22°13'13.5''N, 102°15'55.3''W, Rincón de Romos 22°17'44.9''N, 102°13'06.8''W, and Calera 22°58'39.4''N and 102°41'29.9W, respectively. The sampling carried out was random sampling by conglomerates, dividing each field in groups with plants that showed similar symptoms. The infected plants were stunted in growth, with reddish dying leaves. The stalks and bulbs were soft, and their root system was poorly developed. The collected samples were placed in polyethylene bags and taken to laboratory. The roots and bulbs of 35 plants were cleaned, portions of the diseased tissue was cut into 0.5 cm pieces and disinfected in 1% sodium hypochlorite for 3 minutes. The samples were rinsed twice with sterile distilled water and dried on sterile paper towels. The tissues were cultured on Potato Dextrose Agar (PDA) medium and incubated in the dark at 25°C. Seven days after incubation, pure cultures were obtained using monoconidial cultures technique on Spezieller Nährstoffmmarmer agar (SNA) and subcultured on carnation leaf agar (CLA). Ten isolates were obtained that grew slowly, showing a white coloration, then turning yellow with abundant aerial mycelia. Microscopic traits of 30 characterized spores included slender macroconidia that were curved dorsiventrally, tapering towards both ends, with five to seven thin septa, measuring 36.4-56.6 µm × 4.0-4.9 µm in size and chlamydospores that were abundant, globose to oval, subhyaline and terminal or intercalary in chains measuring 8.8-4.5 µm in diameter. Microconidia, were single-celled, hyaline, nonseptate, and ovoid. The morphological traits matched the description of Fusarium clavum (Xia et al. 2019). To confirm the strain's identity, DNA was extracted from six monoconidial cultures and used as template to amplify translation elongation factor (TEF) gene 1α, RNA polymerase largest subunit (RPB1), and RNA polymerase second largest subunit (RPB2) (O'Donnell et al. 2010). The products were sequenced and deposited in GenBank as ON209360, OM640008 and OM640009, the homology analysis using BLASTn was similar to F. clavum with 99.46%, 99.49% and 98.82% respectively with E VALUE 0.0 in all cases with access numbers OP48709, HM347171 and OP486686. Koch postulate was performed to confirm the pathogenicity of the six isolates. Variegated garlic cloves were planted after being disinfected with sodium hypochlorite at 3% w/v in 2-kg pots under the greenhouse conditions. When the garlic plants developed 4 or 5 true leaves, their basal stalks were inoculated by pouring uniformly with 1 mL of a spore suspension at 108 conidia/mL prepared from 1-week-old colonies (Lai et al. 2020). Twenty-four plants were inoculated with six isolates (four plants per isolate), and four control plants were treated with sterile distilled water. Symptoms appeared 20 days post-inoculation. The leaves were reddish, and the stalks were soft. The leaves eventually developed foliar dieback disease symptoms, their root system showed brown lesions and rot, and all water-inoculated controls remained asymptomatic. Isolations were made on the diseased plants, and the inoculated pathogen was recovered and confirmed morphologically and molecularly by DNA extraction and PCR reactions. Koch's postulate was repeated twice, obtaining the same results. To our best knowledge, this is the first report of F. clavum infecting Allium sativum L. in Mexico. bulb rot caused by F. clavum is a severe threat to garlic cultivation, and identification of this pathogen is important for effective disease management and control.

Saffron Bulb Mite Rhizoglyphus robini (Claparede) Its Occurrence, Biology and Chemical Intervention Tactics on Saffron Corms in Kashmir, India

Saffron corms are attacked by a wide variety of arthropod pest species and mites are one of them. Among Arachnida class bulb mite causes considerable damage to saffron crop and presence of this pest is a regular feature in its cultivation. To record the occurrence of saffron bulb mite, field study was conducted at different saffron growing areas of Kashmir. For biology of the pest, laboratory study was conducted at Department of Entomology, Faculty of Horticulture, Sher-e-Kashmir University of Agricultural Sciences and Technology, Shalimar. The mean percent infestation of saffron bulb mite from all the locations revealed highest and lowest percent mite infestation of 25.83 and 12.50 recorded from Dhussu and Saffron research station respectively. Passing through five active stages viz., Egg, Larva, Protonymph, Tritonymph and adult completed the life cycle of Rhizoglyphus robini. The heteromorphic deutonymph or hypopal stage is sometimes produced for phoresis when population is overcrowded, but this stage was not observed during the study. The results revealed that the females laid their eggs on the corms. The average incubation period of egg was observed to be of 6.16 ±0.78 days while as other developmental periods Larva, Protonymph, and Tritonymph were of 5.22 ±0.68, 5.76 ±0.52, and 7.00 ±0.91 days at 25± 50C respectively. The longevity of adult male and female were 27.9 ±0.44 and 22.9 ±0.69 days respectively. The efficacy of different doses of acaricides against Saffron bulb mite revealed that drenching the soil with chlorpyrifos 20 EC + foliar spray with cyenopyrafen 30 SC showed best results with 4.23 percent leaves with stunted growth and chlorosis, 97.34 percent flower emergence and 2.23 percent bulb rot.

First Report of Pantoea agglomerans Causing a Leaf Blight and Bulb Rot of Onion in Chile.

Onion (Allium cepa L) is an important vegetable crop in Chile with 3.675 ha (ODEPA, www.odepa.gob.cl). Since 2017, a series of symptoms has been increasingly observed, mainly in mature bulbs which include water-soaked lesions on mature leaves and external and central cataphylls, which evolve to necrotic lesions and rot, reducing yields and affecting the postharvest life. As the season progresses over 90% incidence has been estimated in severely affected fields. Symptomatic plants were collected for disease identification in summer (February 2022) in Santa Cruz, O´Higgins, Chile, (34º41'32"S and 71º27'17"W). Isolations were done on semi-selective OEM medium (Zaid et al. 2012) and isolated colonies were transferred and maintained on King's B (KB) medium. Colonies on OEM medium were yellow slightly green in the center, circular to irregular shape, elevated, smooth margin, and mucoid texture (Tho et al. 2015). On KB medium colonies were yellow cream, slightly darker in the center, with smooth margins and circular or slightly irregular. Two isolates (CHS B3, CHS B1), obtaining in three different onion fields, were further identified by PCR amplification of partial 16S rRNA using primers 27F/1492R (Frank et al., 2008). The resulting sequencing were deposited in GenBank under accession numbers OP934962.1 and OP934961.1. A BLAST search of GenBank showed that these sequences exhibited 99,89 to 100 % nucleotides similarities with multiple Pantoea agglomerans sequences. Three isolates were used to test pathogenicity on detached cataphylls and onion plants tests in the laboratory and greenhouse, respectively. Cataphylls (n=10) were surface disinfected with 2% NaOCl for 2 min, placed in sterile Petri dishes, and injected with 50 µL (106 UFC/mL) inoculum suspension with the aid of a hypodermic syringe. Cataphylls were incubated in humid chamber at 22 + 2ºC for 14 days. Additionally, three-month-old onion plants (n=10), growing in pod, were inoculated by injecting leaves with 1 mL inoculum suspension (106 UFC/mL). Plant were incubated under greenhouse for 30 days to 28ºC. An equal number of cataphylls and onion plants, injected with sterile water, were left as controls. Water soaked lesions on inoculated cataphylls appeared five days after inoculation. While onions plants develop water soaked lesion 1- 4 days after inoculation. Blight and bulb rot were observed only on inoculated plants. Symptoms were identical to those previously observed in the field. Reisolations were accomplished for 100% of the inoculated cataphylls and leaves fulfilling Koch's postulates (Biochemical characterization and PCR amplification of partial 16S rRNA). Previously, Pectobacterium carotovorum was reported causing a similar disease in Chile (Acuña 2008). However, to our knowledge this is the first report of P. agglomerans causing onion leaf blight and bulb rot in O'Higgins Region, Chile. In agreement with other reports (De Armas et al. 2015, Tho et al. 2015), leaf blight and bulb has been a very aggressive disease in Chile. These results do not exclude the possibility that other Pantoea species (Armas et al. 2015) may be involved in the leaf blight and bulb rot of onions.

Evaluating Options to Increase the Efficacy of Biocontrol Agents for the Management of Pantoea spp. Under Field Conditions.

Center rot of onion is caused by a complex of plant pathogenic Pantoea species, which can lead to significant yield losses in the field and during storage. Conventional growers use foliar protectants such as a mixture of copper bactericides and an ethylene-bis-dithiocarbamate (EBDC) fungicide to manage the disease; however, organic growers have limited management options besides copper-protectants. Biocontrol agents (BCAs) provide an alternative; however, their efficacy could be compromised due in part to their inability to colonize the foliage. We hypothesized that pretreatment with peroxide (OxiDate 2.0: a.i., hydrogen peroxide and peroxyacetic acid) enhances the colonizing ability of the subsequently applied BCAs, leading to effective center rot management. Field trials were conducted in 2020 and 2021 to assess the efficacy of peroxide, BCAs (Serenade ASO: Bacillus subtilis and BlightBan: Pseudomonas fluorescens), and an insecticide program (tank mix of spinosad and neem oil) to manage center rot. We observed no significant difference in foliar area under the disease progress curve (AUDPC) between the peroxide pretreated P. fluorescens plots and only P. fluorescens-treated plots in 2020 and 2021. Peroxide pretreatment before B. subtilis application significantly reduced the foliar AUDPC as compared with the stand-alone B. subtilis treatment in 2020; however, no such difference was observed in 2021. Similarly, peroxide pretreatment before either of the BCAs did not seem to reduce the incidence of bulb rot as compared with the stand-alone BCA treatment in any of the trials (2020 and 2021). Additionally, our foliar microbiome study showed comparatively higher P. fluorescens retention on peroxide pretreated onion foliage; however, at the end of the growing season, P. fluorescens was drastically reduced and was virtually nonexistent (<0.002% of the total reads). Overall, the pretreatment with peroxide had a limited effect in improving the foliar colonizing ability of BCAs and consequently a limited effect in managing center rot.

First report of leaf rot on Lonicera japonica caused by Rhizopus arrhizus in China.

Lonicera japonica is a perennial shrub that has been used since ancient times as a medicine to clear heat and detoxify poisons. Its branches (the vine of L. japonica) and unopened flower buds (honeysuckle) can be used as medicine to treat external wind heat or febrile disease fever (Shang, Pan, Li, Miao, & Ding, 2011). In July 2022, a serious disease was observed in L. japonica individuals planted in an area of experimental base of Nanjing Agricultural University (N 32°02', E 118°86'), Nanjing, Jiangsu Province, China. More than 200 Lonicera plants were surveyed, and the incidence of leaf rot in Lonicera leaves was over 80%. The initial symptoms were of chlorotic spots and gradual development of visible white mycelia and powdery substances (fungal spores) were observed on the leaves. Both the front and back of the leaves gradually appeared as brown diseased spots. Thus, a combination of multiple disease spots causes leaf wilting and the leaves eventually fall off. Leaves with typical symptoms were collected and cut into approximately 5 mm square fragments. The tissues were sterilized in 1% NaOCl for 90 s and 75% ethanol for 15 s and then washed with sterile water three times. The treated leaves were cultured on Potato Dextrose Agar (PDA) medium at 25℃. When mycelia grew around the leaf pieces, fungal plugs were collected along the outer edge of the colony and transferred to fresh PDA plates using a cork borer. Eight fungal strains with the same morphology were obtained after three rounds of subculturing. The colony was initially white with a fast growth rate, and occupied a 9-cm-diameter culture dish within 24 h. The colony turned gray-black in the later stages. After 2 days, small black sporangia spots appeared on top of the hyphae. The sporangia were yellow when immature, and black at maturity. The spores were oval with an average size of 29.6 (22.4-36.9) × 35.3 (25.8-45.2) µm (n = 50) in diameter. To identify the pathogen, fungal hyphae were scraped, and the fungal genome was extracted using a kit (BioTeke, Cat#DP2031). The internal transcribed spacer region (ITS) of the fungal genome was amplified with primers ITS1/ITS4, and the results of ITS sequencing were uploaded to the GenBank database with accession number OP984201. The phylogenetic tree was constructed using the neighbor-joining method with MEGA11 software. Phylogenetic analysis based on ITS showed that the fungus was grouped together with Rhizopus arrhizus (MT590591) and had high bootstrap support. Thus, the pathogen was identified as R. arrhizus. To verify Koch's postulates, 60 ml of a spore suspension (1×104 conidia/ml) was sprayed onto the surface of 12 healthy Lonicera plants, and the other 12 plants were sprayed with sterile water as a control. All plants were kept in the greenhouse at 25°C with 60% relative humidity. After 14 d, the infected plants showed symptoms similar to those of the original diseased plants. The strain was isolated again from the diseased leaves of artificially inoculated plants and verified as the original strain by sequencing. The results showed that R. arrhizus was the pathogen responsible for Lonicera leaf rot. Previous studies have shown that R. arrhizus causes garlic bulb rot (Zhang et al., 2022) and Jerusalem artichoke tuber rot (Yang et al., 2020). To our knowledge, this is the first report of R. arrhizus causing Lonicera leaf rot disease in China. Information regarding the identification of this fungus may be helpful for controlling the leaf rot disease.

First report of Fusarium equiseti causing bulb rot on lily (Lilium 'White planet' ) in China.

Lily (Lilium spp.) is one of the main ornamental plants grown in the world. In addition, bulbs of lily have been extensively used as edible and medicinal herbs in northern and eastern Asia, especially in China (Yu et al. 2015; China Pharmacopoeia Committee 2020; Tang et al. 2021). In August of 2021, a disease of stem and leaf rot was observed on lily cultivar 'White planet' with approximately 25% disease incidence in the greenhouse and fields at the Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences (Beijing, China). The bulbs of symptomatic plants were brown and rotten, with sunken lesions. Symptomatic plants showed short, discolored leaves, and eventually lead to stem wilt and death of the whole plants. Infected bulbs were surface sterilized in 75% ethanol for 30 s, then in 2% sodium hypochlorite for 5 min, and rinsed three times with sterile distilled water. A 0.5×0.5 cm2 tissue piece was then placed on potato dextrose agar (PDA) medium and incubated at 25±1℃. After 5 days, the isolate was purified by single spore isolation technique. The singled-spored fungal colony was characterized by fluffy white aerial mycelia, and produced orange pigments with age. After seven days on Spezieller Nahrstoffarmer agar (SNA), conidia produced from simple lateral phialides. Macroconidia have pronounced dorsiventral curvature typical, significantly enlarged in the middle, a tapered whip-liked pointed apical cell and characteristic foot-shaped basal cell, 3 to 6 septate, measuring 18.71 to 43.01×2.89 to 5.56 μm with an average size of 26.98×3.90 μm (n=30). Microconidia were not observed. Typical verrucose thick chlamydospore with rough walls were profuse in chains or clumps, ellipsoidal to subglobose. These morphological characteristics were consistent with Fusarium spp. (Leslie et al. 2006). For molecular identification, the internal transcribed spacer (ITS), translation elongation factor subunit 1-alpha (TEF1-α) and RNA polymeraseⅡsubunit 2 (RPB2) genes were amplified using primers ITS1/ITS4, EF1/EF2 and 5F2/7cR respectively and sequenced (White et al. 1990; Jiang et al. 2018; O'Donnell et al. 2007). Sequences were submitted to GenBank under accession numbers OM078499 (ITS), Accession OM638086 (TEF1-α) and OM638085 (RPB2). BLAST analysis showed that ITS, TEF1-α and RPB2 sequences shared 100%, 99.8%, 99.2% identity to F. equiseti (OM956073, KY081599, MW364892) in GenBank, respectively. In addition, ITS, TEF1-α and RPB2 sequences shared 100%, 99.53%, 100% identity with Fusarium lacertarum (LC7927, Fusarium incarnatum-equiseti species complex) in the Fusarium-ID database. Based on the morphological characteristics and molecular sequences, the isolates were identified as Fusarium equiseti. A pathogenicity test was performed on potted lily ('White planet') under greenhouse conditions (25±1℃ with a 16 h light and 8 h dark cycle). Three healthy lily bulbs were selected and one bulb was planted in each pot filled with sterilized soil. Each pot was inoculated with 5 mL of conidia suspension (1×107 conidia/mL) in te soil around bulbs with a stem length of 3 cm, with an equal amount of sterilized water as a control. This test had three replicates. After 15 days of inoculation, typical symptoms of bulb rotten, like those observed in the greenhouse and fields, developed on the inoculated plants but not on the controls. The same fungus was consistently reisolated from the diseased plants. To our knowledge, this is the first report that F. equiseti caused bulb rot on Lilium in China. Our result should help with future monitoring and control of lily wilt disease.

Cultural Management Strategies to Reduce the Risk of Onion Bacterial Diseases

AbstractOnion production in the United States is massive in scale: 3.5 million tons of onions were produced on 130,000 acres and valued at approximately $1 billion, on average, each year from 2018 to 2022. Unfortunately, this highly valuable crop can be threatened by plant pathogenic bacteria that cause a variety of bacterial leaf blights and bulb rots, diseases for which there is no cure. An integrated approach is needed that emphasizes multiple preventative cultural management strategies to reduce the risk posed by onion bacterial diseases. Earn 0.5 CEUs in Integrated Pest Management by reading this article and taking the quiz at https://web.sciencesocieties.org/Learning‐Center/Courses.

First Report of Neck Rot Caused by Botrytis aclada on Onion (Allium cepa L.) in Gansu Province, China.

Onion (Allium cepa L.) is one of the most important cultivated vegetable crops in many countries, including China (Hanci 2018; Steentjes et al. 2021). Gansu, in the northwest region of China, is a major area of onion production (Zhang et al. 2022). In October 2021, typical symptoms of neck rot were observed on stored onion bulbs (cv. Honghe) in Jiuquan, Gansu Province, China. Further surveys of 20 bags of onion bulbs randomly selected in a storage facility with bulbs harvested from 73 ha indicated that approximately 5% of the bulbs had typical neck rot symptoms. The neck of infected bulbs developed a water-soaked decay, with softened and discolored inner scales with white to gray mycelium in the neck. Infection usually began in the neck and sometimes spread through the entire bulb. In severely affected bulbs, the fleshy scales were span style="font-family:'Times New Roman'">decayed and the bulbs had shrunk, with black sclerotia between the rotting scales in the neck and shoulders of the bulb. Small pieces cut from the margins of lesions were surface-disinfested with 75% ethanol for 10 s, and 1% NaClO for 1 min, rinsed three times in sterile distilled water, dried on sterile filter paper, placed onto potato dextrose agar (PDA), and incubated at 20 ± 1℃ for 5 days in the dark. Ten pure cultures were obtained by single-spore isolation (one from each of 10 bulbs). All 10 isolates produced colonies that initially were white, and became gray to dark in color with gray mycelium that was covered with abundant conidia resembling Botrytis species when cultured on PDA at 20 ± 1℃ for 10 days in the dark. Conidia (n = 100 per isolate) were one-celled, ellipsoid or ovoid, beige to dark brown, and 5.6 to 14.8 × 3.9 to 8.5 μm. Sclerotia were not produced by any of the isolates on PDA after incubation at 20 ± 1°C for 30 days. The internal transcribed spacer (ITS) region of ribosomal DNA, glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene, heat shock protein 60 (HSP60) gene, and DNA-dependent RNA polymerase II core subunit (RPB2) gene of two representative isolates, JQ21AC03 and JQ21AC09, were amplified and sequenced with primers ITS1/ITS4, G3PDH-F/G3PDH-R, HSP60-F/HSP60-R, and RPB2-F/RPB2-R, respectively (Staats et al. 2005), and deposited in GenBank (ITS: OP604277 and OP604283; G3PDH: OP627512 and OP627515; HSP60: OP627513 and OP627516; and RPB2: OP627514 and OP627517). BLASTn analysis of the resulting sequences showed 98 to 100% similarity with those of Botrytis aclada. A maximum likelihood phylogenetic tree generated by combining the sequenced loci using MEGA11.0, clustered isolates JQ21AC03 and JQ21AC09 with B. aclada with 99% bootstrap support. Based on these results, isolates JQ21AC03 and JQ21AC09 were identified as B. aclada. Pathogenicity of the two isolates on the onion cv. Honghe was confirmed by inoculating 30 heathy onion bulbs per isolate with 100 μL of conidial suspension (1×105 conidia/ml), while control bulbs were inoculated with 100 μL sterile distilled water. All 30 bulbs inoculated with each isolates developed neck rot symptoms after 30 days incubation at 15 ± 1ºC in the dark, with symptoms identical to those observed in the original storage facility, whereas control bulbs remained symptomless. The pathogen was re-isolated from the symptomatic tissue of inoculated bulbs, fulfilling Koch's postulates. hang et al (2008) documented this pathogen causing bulb rot of onions in Wuhan, Hubei Province, 1,700 km from Gansu. To the best of our knowledge, this is the first report of B. aclada causing neck rot on onion bulbs in storage in Gansu Province, which is the main onion production region in China. Considering that onion is the main source of income for growers in Gansu, further studies will be required to understand the epidemiology of this disease and foster appropriate disease management measures to avert disease outbreaks in the future.

Firstreportflat peach fruit rotcausedbyFusariumfujikuroi in China.

Flat peach (Prunus persica L. Batsch. var. compressa Bean) is a distinctive peach variety with flat shape. It is well known for its high nutritional value and pleasant flavor, and has become the primary cultivar in Shengzhou city, Zhejiang province, China. In July 2021, we discovered that 8%-10% of flat peach (data gained from a survey of fifteen trees in each orchard) fruits in three orchards (about 2.6 ha) had a fungal disease in Jinting Town, Shengzhou. The infected fruit by this pathogen showed soft, brown, and sunken lesions, accompanied by a sour odor and white mycelia. For pathogen isolation, ten infected fruits were surface sterilized (75% ethanol for 30 s and 2.0% NaClO for 3 min), then rinsed with sterile distilled water three times. The tissues from the margin of lesions were cut into 0.6×0.6 cm pieces and transferred to potato dextrose agar (PDA) medium containing 30.0 μg/mL chloramphenicol. The plates were incubated at 25 °C for 5 days in the dark. Mycelia growing from tissues were subcultured onto fresh PDA medium to get a pure isolate, which formed dense white hyphae after 4 days. The average growth rate of mycelium on PDA medium was 3.4 ± 0.2 mm/d, and the colonies were pale purple after 5 days. Macroconidia was slender, slightly curved, almost 2 to 3 septa, with a bend and tapering apical cell and poorly developed foot cell, and the size was 4.5 to 16.9 × 1.8 to 4.0 μm (n= 50) μm. No microconidia and chlamydospores were observed. This isolate's morphological and cultural characteristics were close toFusarium fujikuroi(Leslie and Summerell. 2006). To further get the phylogenetic evidence, the nuclear ribosomal internal transcribed spacer region (ITS) and translation elongation factor-1 (TEF-1α) genes of three representative isolations were amplified by primers ITS1 (F:5'-GGAAGTAAAAGTCGTAACAAGG-3') /ITS4 (R5'-TCCTCCGCTTATTGATATGC-3') (Groenewald et al. 2013), and EF1-728F (5'-TACAARTGYGGTGGTATYGACA-3')/ EF1-986R (5'-ACNGACTTGACYTCAGTRGT-3') (Carbone and Kohn. 1999), respectively. The amplified sequences were submitted to Genbank with accession numbers OP223318 for the IST and OP394152 for the ETF-1α region. Blastn results indicated that the ITS and TEF-1α sequences from the isolated strain shared 96.25% and 98.32% similarity with two strains of F.fujikuroi (GenBank Accession No. MF281286.2, and MK311296.1). The isolates were clustered with F.fujikuroi clade (Supplementary Fig.1 and Supplementary Fig.2), consistent with the morphological identification. To evaluate Koch's postulates, 20μL of spore suspension (1×106 spores/mL) were inoculated into ten healthy flat peach fruits with sterile syringes, while another ten healthy fruits were injected with sterilized water as controls. All fruits were kept in sealed plastic boxes at 25 °C with 90% relative humidity. All inoculated fruits showed symptoms similar to those of naturally infected fruits, while uninoculated fruits remained healthy after 4 days. In addition, F. fujikuroi was an important pathogen causing bakanae disease in rice (Hou et al. 2017). This pathogen was also reported to cause Lilium lacifolium Thunb bulb rot (Fang et al. 2022) and cause root rot in peanuts in China (Sun et al. 2022). However, to the best of our knowledge, this is the first record of F. fujikuroi causing fruit rot in flat peach in China. The discovery will provide helpful information about flat peach rot disease management.

Occurrence, Identification and Preliminary Biological Control of Bulb Rot of Onion (Allium cepa)

Onion bulb rot was observed in low-temperature warehouses in Mae Wang District, Chiang Mai, Thailand in 2022. The pathogens were isolated and identifi ed as Pantoea agglomerans and Burkholderia gladioli based on morphological and biochemical characterization, pathogenicity tests, and 16S rRNA gene sequences. Biological control is regarded as a safer and more sustainable strategy than chemical control and has elicited great attention for disease control. Bacillus sp. isolate BB22 was isolated from rhizosphere soil and was evaluated for its preliminary potential to inhibit the growth of onion bulb rot bacterial pathogens by the dual culture technique. The results showed inhibition zones of P. agglomerans of 30.1 mm and B. gladioli of 13.0 mm. Molecular identifi cation of Bacillus sp. isolate BB22 was performed by using 16S rRNA and gyrA gene sequences and confi rmed as Bacillus velezensis. PCR amplifi cation of the bmyB, fenD, ituA, and srfAA genes coding for bacillomycin, fengycin D, iturin A, and surfactin, respectively, showed the potential for the production of these antibiotics. This study revealed the potential application of B. velezensis as a biocontrol agent against bulb rot-causing bacterial pathogens of onions.

Assessment of the planting material infection and fungal pathogens identification of garlic

The research results of the main diseases that cause garlic bulb rot in the Moscow region during storage are presented. It was shown that long-term storage (17 months), as expected, increased the rots prevalence and development, both Penicilus: 10.0-12.5%, and Fusarium: 10.0-11.3%. Winter garlic cultivar Nina Arsentyevna’s Memory bulbs by 2 species of Fusarium hade been infected: F. fujikuroi and F. roliferatum identified by PCR analysis and subsequent sequencing of the two isolates target genome. Winter garlic cultivar Pamyati Nina Arsentievna is highly resistant to rots of fungal origin, even after long-term storage (more than a year), in comparison with the reference sample from the Federal Scientific Center for Vegetable Growing, the bulbs of which were completely affected, and corresponded to the fifth defeat point. On winter garlic, the causative agent of penicillium rot, Penicillium glaucum Link, was also found. On collection samples of winter garlic, during selective annual studies, penicillary rot was not found.

Aphelenchoides varicaudatus (Nematoda: Aphelenchoididae) and Helicotylenchus erythrinae (Nematoda: Hoplolaimidae) from Garlic Plantation in Magelang, Central Java, Indonesia.

There were two most found genera of plant parasitic nematodes from garlic plantation in Magelang, Central Java, Indonesia which suffered losses due to bulb rot, Aphelenchoides and Helicotylenchus. Polymerase Chain Reaction (PCR) was conducted using a pair of universal nematode primer (D2A/D3B) to determine the Aphelenchoides and Helicotylenchus species from those host. Both genera were amplified at ~780 bp. The Blast-N results for the Aphelenchoides showed high identity to Aphelenchoides varicaudatus from Yunnan China (HQ283353) with 99.47 % identity, while the Helicotylenchus showed 95.22 % identity to Helicotylenchus erythrinae from Colombia (MT321739). From morphological and molecular data, we confirm that the Aphelenchoides species is A. varicaudatus. Based on female morphological character, Helicotylenchus species refers to H. erythrinae. Which is also supported by its nucleotide alignment which has same region character as H. erythrinae (MT321739). This is the first report of molecular characterization of H. erythrinae in Indonesia.

Reducing the risk of onion bacterial diseases: A review of cultural management strategies

AbstractOnion (Allium cepa L.) bacterial diseases can cause catastrophic crop losses. Chemical and biological control products are not curative and have mixed or limited efficacy at preventing onion bacterial diseases. Cultural management strategies, such as appropriate irrigation and nitrogen (N) management practices, can reduce the risk for bacterial infection and disease progression. This review synthesizes available literature on cultural management strategies to mitigate onion bacterial diseases and identifies knowledge gaps, promising strategies, and limitations. This synthesis revealed that high N fertilizer application rates, the use of overhead irrigation, and late‐season irrigation or rainfall, especially during field curing, can increase bacterial disease pressure. These factors contribute to conducive conditions for bacteria to colonize the foliage and neck of onion plants. Other in‐season strategies, such as reducing the total amount of irrigation or adjusting application rates of nutrients other than N, have not limited losses to bacterial bulb rots while maintaining desired yields. At the end of the growing season, appropriate timing of lifting bulbs (undercutting to sever the roots) and other practices that speed field curing can reduce the risk of bacterial bulb rots. However, it is less clear if topping (removing foliage) before or after curing influences bacterial bulb rots. Conflicting results reported for some cultural management strategies indicate that local conditions, such as climate and soil types, must be considered for optimizing practices to manage onion bacterial diseases. Advancing our scientific understanding and grower awareness of strategies that reduce onion bacterial diseases is a cost‐effective opportunity to avoid losses.

First Report of Fusarium acuminatum Causing Leaf Blight on Garlic in China.

In June 2021, leaf blight symptoms were detected on garlic plants (Allium sativum) in southeastern Jiangsu (Nantong municipality; 120.61° E, 33.25° N) in China. Two-month-old garlic plants exhibited leaf tip die back and light brown lesions in new and old leaves (Figure 1). The symptoms were observed in 40% of the plants in a 60-square-meters commercial field surrounded by rice fields, and were similar to those reported for Botrytis porri, Septoria allii and Stemphylium eturmiunum causing leaf blight on garlic (Dumin et al. 2021; Park et al. 2013; Zhang et al. 2009). Six samples of symptomatic tissue collected in Nantong municipality, approximately 1 cm2 in size, were sterilized in 2% NaOCl for 15 min and washed twice with sterile ddH2O. The pathogen was isolated from all collected samples on PDA medium, containing 50 µg/mL chloramphenicol, at 26°C. Pink colonies with orange pigmentation were observed after 7 days. Internal transcribed spacer (ITS), elongation factor 1-α (EF1-α), RNA polymerase II largest subunit (RPB1) and RNA polymerase II second largest subunit (RPB2) genes were amplified using ITS1/ITS4, EF1-728F/EF1-986R, RPB1-R8/RPB1-F5 and fRPB2-7CF/fRPB2-11aR primers, respectively. A total of 17 isolates were obtained, with nine of the isolates sharing the same sequences (strain NJC21), six of the isolates sharing the same sequences (strain NJC22), and the other two isolates showing different sequences (strains NJC23 and NJC24). The obtained sequences were submitted in GenBank under accession numbers OL655398-OL655401 (ITS), and OL741712-OL741723 (EF1-α, RPB1, RPB2). The obtained ITS sequences shared >99% homology to the ITS gene from F. acuminatum IBE000006 (EF531232), the EF1-α sequences shared 99% homology to the EF1-α gene from F. acuminatum F1514 (LC469785), the RPB1 sequences shared >99% homology to the RPB1 gene from F. acuminatum JW 289003 (MZ921675), and the RPB2 sequences shared 100% homology to the RPB2 gene from F. acuminatum NL19-077002 (MZ921813) or 100% homology to the RPB2 gene from F. acuminatum MD1 (MW164629). A phylogenetic tree was constructed using MEGA7 with related Fusarium strains (Figure 2). Microscope observations after incubation in potato-sucrose-agar (PSA) medium showed the presence of oval microconidia, fusiform macroconidia, septate mycelium and chlamydospores, and agree with the morphology of F. acuminatum (Marek et al. 2013). The pathogenicity was screened with two-week-old wounded and non-wounded garlic plants using a 1 × 106 spores/mL solution (20 µL). Sterile ddH2O was used in the control experiment. The inoculated plants were incubated at 26°C and 60% relative humidity for 3 days, detecting similar lesions compared to those observed in the field. The pathogen was recovered from 5 different lesions, from different plants, and its identity was confirmed by sequence analysis. Recently, F. acuminatum was reported to cause garlic bulb rot in Serbia (Ignjatov et al. 2017). Although F. acuminatum is well known as a causal agent of root rot (Li et al. 2021; Tang et al. 2021), F. acuminatum has also been found causing leaf blight on onion (Parkunan et al. 2013) and muskmelon (Yu et al. 2021). This is the first report of F. acuminatum causing leaf blight on garlic, demonstrating the host and tissue promiscuity of this pathogen. China is the largest producer of garlic in the world with nearly 20 million tons harvested in 2020. This report will help to better understand the pathogens that are affecting garlic production in China.

Storability study of different short-day onion (Allium cepa L.) genotypes/varieties

A total of 25 short-day onion genotypes along with two recommended varieties viz. BARI Piaz-1 and BARI Piaz-4 were evaluated for 5 and ½ months at the bamboo-made ambient storage. The study was conducted at Spices Research Sub-Centre, Bangladesh Agricultural Research Institute, Faridpur, Bangladesh in 2020 with the geographic coordinates- 23̊ 11̍ N and 89̊ 09̍ E. for assessing their storability. All genotypes in the trial were evaluated under 25 accession lines such as AC Bog 409, AC Bog 410, AC Bog 411, AC Bog 412, AC Bog 413, AC Bog 414, AC Bog 415, AC Bog 416, AC Bog 417, AC Bog 419, AC Bog 420, AC Bog 421, AC Bog 422, AC Bog 423, AC Bog 424, AC Bog 425, AC Bog 426, AC Bog 427, AC Bog 428, AC Bog 429, AC Bog 430, AC Bog 431, AC Bog 433, AC Bog 434 and AC Bog 435. The experiment was laid out in a completely randomized design with three replications. The storage data were recorded periodically at 20 days interval (20 May, 08 June, 27 June, 16 July, 04 August, 23 August, 11 September, 30 September and 19 October) on rotting (%), sprouting (%), physiological loss in weight (PLW, %) and total loss (%) of bulbs. The results revealed that the genotypes/varieties had significant difference on the rotting, sprouting, PLW and total loss of bulbs. The genotype AC Bog 430 showed the lowest cumulative rotting loss (10.47%) but AC Bog 429 exhibited the highest cumulative rotting loss (39.47%). The maximum and minimum cumulative sprouting losses were recorded from AC Bog 434 (23.03%) and AC Bog 429 (1.47%). The BARI Piaz-1 had the lowest cumulative PLW (19.39%). However, the AC Bog 420 had the highest cumulative PLW (44.68%). The highest and the lowest cumulative total losses were calculated from the AC Bog 433 (79.99%) and AC Bog 430 (44.62%). The rate of storage loss was higher in May to July than that of August to October. The rotting, sprouting, PLW and total losses were increased with the increase of the onion storage period. The average rotting, sprouting, PLW and total losses were 20.71%, 13.71%, 29.87% and 64.29%, respectively for 5 and ½ months storage of onion under the study. The storage capacity was not correlated with the skin colour of onion. The dry matter content of bulb was found partial associated with the storage capacity. Finally, under the present storability study, AC Bog 430 (44.62%), AC Bog 426 (45.28%), AC Bog 413 (47.13%) and BARI Piaz-1 (45.31%) showed good performance on the basis of total weight loss.

Rouxiella badensis, a new bacterial pathogen of onion causing bulb rot.

Onion bulb rot can be caused by multiple plant pathogens including bacterial pathogens. During our routine survey of commercial onion farms in 2014, 2020, and 2021, seven putative Rouxiella spp. strains were isolated from symptomatic onion samples in Georgia, United States. Upon fulfilling Koch's postulates on onion, a genome analysis was conducted. Whole-genome indices (ANI and dDDH) showed that the strains belonged to Rouxiella badensis. Although the seven R. badensis strains were not pathogenic on onion foliage, the strains were able to cause bulb rot and could also produce necrotic lesions in a red onion scale assay. R. badensis populations increased significantly and to a level comparable to P. ananatis PNA 97-1R in a red onion scale infection assay. The core-genome analysis grouped all onion R. badensis strains from Georgia together, and distinct from R. badensis strains isolated from other sources and locations. Based on the genome analysis of strains (from the current study and available genomes in the repository), type I, III (Ssa-Esc and Inv-Mxi-Spa types), and V secretion systems are present in R. badensis genomes, while type II, IV, and VI secretion systems are absent. However, various secondary metabolite gene clusters were identified from R. badensis genomes, and a thiol/redox-associated enzyme gene cluster similar to the Pantoea alt cluster mediating thiosulfinate tolerance was also present in onion strains of R. badensis. This is the first report of R. badensis as a plant pathogen.