First Report of Berkeleyomyces basicola Causing Black Root Rot on Lisianthus (Eustoma grandiflorum) in China

Lisianthus (Eustoma grandiflorum (Raf.) Shinn.) is an important ornamental plant ranking in the top 10 cut flowers worldwide (Xiao et al., 2018). In 2020 and 2021, black root rot was found as a major disease limiting lisianthus production in Yunnan Province, China. Black root rot was first observed in early July 2020 on lisianthus grown in a commercial flower-production plantation, with nearly 60% plants infected. Symptoms appeared as coalescing necrotic lesions leading to black discoloration of the roots. Root damage induced by disease resulted in insufficient water and nutrient uptake by the plant, causing stunting and whole-plant wilting. The pathogen could not infect the intact endodermis, and vascular tissues below the discolored cortical tissue remained healthy. Symptomatic roots were surface sterilized using 1% NaClO for 1 min, rinsed three times in sterile water, placed onto potato dextrose agar (PDA), and incubated at 25°C for 7 days in the dark. The morphological characteristics were basically consistent: the colonies were white to gray in color, and the conidiophores were colorless to brown, solitary or clustered. Conidia were single-celled, colorless, rod-shaped, and obtuse at both ends. Chlamydospores were dark brown, clustered or solitary. The morphological characteristics of the pathogen were similar to those of Berkeleyomyces basicola (Berk. & Broome) W.J. Nel, Z.W. de Beer, T.A. Duong & M.J. Wingf. (Nakane et al. 2019). DNA was extracted from mycelia of representative isolate TB using the Plant Genomic DNA Kit (Tiangen, Beijing, China). The internal transcribed spacers (ITS), DNA replication licensing factor (MCM7), ribosomal large subunit (LSU), and 60S ribosomal protein RPL10 (60S) regions were amplified with primer pairs ITS1/ITS4 (Groenewald et al. 2013), MCM7-for/MCM7-rev, LR0R/LR5, and 60S-506F/60S-908R, respectively (Nel et al. 2018). Phylogenetic analysis of multiple genes (Bakhshi et al. 2018) was conducted with the maximum likelihood method using MEGA7. The sequences of our isolate (TB) and three published sequences of B. basicola were clustered into one clade with a 100% bootstrapping value. The accession numbers of B. basicola reference sequences are MF952423 (ITS), MF967079 (MCM7), MF948658 (LSU), and MF967072 (60S) of isolate CMW6714; MF952428 (ITS), MF967088 (MCM7), MF948661 (LSU), and MF967073 (60S) of isolate CMW25440; MF952429 (ITS), MF967102 (MCM7), MF948659 (LSU), and MF967075 (60S) of isolate CMW49352. The sequences of TB have been deposited in GenBank with accession numbers MZ351733 for ITS, MZ695817 for MCM7, MZ695816 for LSU, and MZ695815 for the 60S region. To verify the pathogenicity of the fungus, inoculations were performed on ten 2-month-old potted lisianthus plants by dipping the roots into a conidial suspension (105 spores/ml) for 2 h. Ten plants were mock inoculated with distilled water as a control. Symptoms of black root rot were observed 30 days after inoculation, whereas the control roots remained healthy. The causal fungus has a host range of over 230 species and is a destructive pathogen of many crops and ornamental plants, including cotton (Gossypium barbadense L.), tobacco (Nicotiana tabacum L.) and mango (Mangifera indica L.) (Shukla et al. 2021; Toksoz and Rothrock 2009). This is the first report worldwide of B. basicola infecting lisianthus. This discovery is of great importance for Chinese flower growers because this fungus is well established in the observed area, and effective measures are needed to manage this disease.

First report of Berkeleyomyces basicola (synonymous: Thielaviopsis basicola) on roots of sweet potato (Ipomoea batatas (L.) Lam) in Argentina

Symptomatic sweet potato cv Arapey INIA samples were collected from a commercial production field in Colonia Molina, Guaymallén department, Mendoza province, Argentina. They showed dark rounded lesions, sometimes coalescing with white granular mycelium. Fungus was obtained from symptomatic sweet potatoes, which represented the generalized infection that affected the crop. They were seeded in PDA with streptomycin sulfate and incubated for seven days at 21°C, alternating white/black (UV400nm) light. Observations with an optical microscope revealed the presence of hyaline, not septated, cylindrical endoconidia with rounded ends. They were 8-16 μm length and 4–6 μm width. Phialides were 43-46 μm length, rounded bases (7-9 μm width) and tapering to the neck´s tip (4-6 μm width). Brown chlamydospores (aleuriospores), 9-13 μm length and 8-12 μm width, in chains of 2-8 spores were observed. For molecular identification, total genomic DNA was extracted. ITS fragment of 565 pb was amplified using ITS5/ITS4 primers and sequenced. The sequence indicated 99% identity with Berkeleyomyces basicola (synonymous: Thielaviopsis basicola). This was deposited in GenBank as (KX580957) (CBS: C430.74, Gen Bank accession number AF275482.1). This is the first report of B. basicola in sweet potato in Argentina, a potential threat to storage root yields. Highlights: Sweet potato black root rot, new disease in Argentina. First report of Berkeleyomyces basicola causing black root rot on sweet potato in Mendoza, Argentina.

In-vitro compatibility assay of indigenous Trichoderma and Pseudomonas species and their antagonistic activities against black root rot disease (Fusarium solani) of faba bean (Vicia faba L.)

Background Faba bean (Vicia faba L.) cultivation is highly challenged by faba bean black root rot disease (Fusarium solani) in high lands of Ethiopia. To ensure sustainable production of faba beans, searching for eco-friendly disease management options is necessary to curb the progress of the disease timely. The indigenous biocontrol agents that suit local environments may effectively strive with in-situ microorganisms and suppress local pathogen strains. This study aimed to screen antagonistic indigenous compatible Trichoderma and Pseudomonas strains against Fusarium solani. In the pathogenicity test, soil-filled pots were arranged in complete random block design and sown with health faba bean seeds. The effect of some fungicides was evaluated against Fusarium by food poisoning methods to compare with the biocontrol agents. The antagonistic efficacy of biocontrol agents and their compatibility was investigated on Potato dextrose agar medium. Results Fusarium solani AAUF51 strain caused an intense root rotting in faba bean plant. The effect of Mancozeb 80% WP at 300 ppm was comparable with Trichoderma and Pseudomonas strains against Fusarium. The mycelial growth of test the pathogen was significantly (P ≤ 0.05) reduced to 86.67 and 85.19% by Trichoderma harzianum AAUW1 and Trichoderma viridae AAUC22 strains in dual culture, respectively. The volatile metabolites of Pseudomonas aeruginosa AAUS31 (77.78%) found the most efficient in reducing mycelial growth of Fusarium followed by Pseudomonas fluorescens AAUPF62 (71.11%) strains. The cell-free culture filtrates of Pseudomonas fluorescens AAUPF62 and Pseudomonas aeruginosa AAUS31 were more efficient than the Trichoderma strain in reducing the growth of Fusarium isolates. There was no zone of inhibition recorded between Trichoderma harzianum AAUW1, Trichoderma viridae AAUC22, Pseudomonas aeruginosa AAUS31, and Pseudomonas fluorescens AAUPF62 strains, hence they were mutually compatible. Conclusions The compatible Trichoderma and Pseudomonas strains showed antagonistic potentiality that could be explored for faba bean protection against black root rot disease and might have a future dual application as biocontrol agents.

Rosellinia bunodes (black root rot).

Given the difficulty in attributing crop losses solely to R. bunodes, meaningful economic estimates are rare. In coffee (Coffea arabica) in Colombia, losses of $489 ha-1 yr-1 due to Rosellinia have been reported (Bautista and Magdiel, 2000). They were most likely caused by R. bunodes and/or R. pepo. In potato in the Andes, losses due to Rosellinia spp. vary from 20% to 80% and, in prolonged monoculture, can reach 100% (Orellana, 1978).

Characterization and Genetic Mapping of Black Root Rot Resistance in Gossypium arboreum L.

Black root rot (BRR) is an economically important disease of cotton and other crops, especially in cooler regions with short growing seasons. Symptoms include black discoloration of the roots, reduced number of lateral roots and stunted or slow plant growth. The cultivated tetraploid Gossypium species are susceptible to BRR. Resistance to BRR was identified in G. arboreum accession BM13H and is associated with reduced and restricted hyphal growth and less sporulation. Transcriptome analysis indicates that BM13H responds to infection at early time points 2- and 3-days post-inoculation, but by day 5, few differentially expressed genes are observed between infected and uninfected roots. Inheritance of BM13H resistance to BRR was evaluated in an F6 recombinant inbred population and shows a single semi-dominant locus conferring resistance that was fine mapped to a region on chromosome 1, containing ten genes including five putative resistance-like genes.

Application of Bacillus pumilus isolates for management of black rot disease in strawberry

Background Black root rot of strawberry plants caused by Rhizoctonia solani, Fusarium solani, and Pythium sp. is a serious disease in Egypt. Biocontrol agents have frequently proved to possess paramount and safe tools against many diseases. The impact of soil treatments with 3 Bacillus pumilus isolates on black root rot disease of strawberry plants caused by R. solani, F., and Pythium sp. under laboratory and field conditions was examined herein on the commonly used ‘Festival’ strawberry cultivar. To increase the bacterial adhesion and distribution on the roots, each seedling was dipped in bacterial cell suspension at 1 × 108 colony-forming units/ml of each separate bacterial isolate for 30 min then mixed with 5% Arabic gum. Results The tested B. pumilus isolates significantly reduced the growth area of these 3 fungi. The two bacterial isolates Nos. 2 and 3 reduced the growth area by more than 85.2, 83.6, and 89.0% for R. solani, F. solani, and Pythium sp., respectively. Likewise, the 3 bacterial isolates significantly (P ≤ 0.05) inhibited the disease under field conditions. Isolates Nos. 2 and 3 suppressed the disease incidence by 64.4 and 68.9% and disease severity by 65.3 and 67.3%, respectively. The fungicide Actamyl had effect similar to that of the 2 isolates. B. pumilus isolates significantly enhanced growth parameters and yields of strawberry plants; isolates Nos. 2 and 3 raised the yield by 66.7 and 73.3%, respectively. Conclusions Bacillus pumilus isolates could effectively manage the black rot disease in strawberry herein. Due to the significant impact of the root rot disease on strawberry yield, B. pumilus should be further tested to manage the disease on strawberry on large scale in Egypt.