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Biocontrol efficacy of Trichoderma asperellum-enriched coconut fibre against Fusarium wilts of cherry tomato

Hasan ZAE, Mohd Zainudin NAI, Aris A, Ibrahim MH, Yusof MT

J Appl Microbiol, 2020 Oct;129(4):991-1003.
PMID: 32324939 DOI: 10.1111/jam.14674

AIMS: Agro-based wastes were evaluated as a medium for mass micropropagule production and optimal efficacy of Trichoderma asperellum B1092 in controlling Fusarium oxysporum f. sp. lycopersici and promoting tomato growth. This study focused on biological control because pathogen persistence in the soil makes the disease difficult to control.
METHODS AND RESULTS: Rice bran, biochar, empty fruit bunches, coconut fibres, compost, top soil and mixed soil were evaluated as media for mass multiplication of T. asperellum, which is effective in controlling plant pathogens. Yielding the most colony forming units (CFU) among the media, coconut fibre was deemed most suitable for promoting sporulation. After 120 days on the medium, T. asperellum B1902 produced 9·053 × 105 CFU per gram coconut fibre; oil palm empty fruit bunches was second highest (7·406 × 105 CFU per gram). In field tests of T. asperellum B1092 against F. oxysporum f. sp lycopersici (causing Fusarium wilt of cherry tomato), B1092 significantly promoted plant growth compared to the control. The efficacy of this formulation resulted in increased growth of roots and shoots tomato plants and total lycopene, sugar, K, N, Ca, P and Mg content after 120 days.
CONCLUSIONS: Trichoderma asperellum B1092 showed great field potential for improving productivity and quality of tomatoes and in controlling Fusarium wilt of cherry tomato.
SIGNIFICANCE AND IMPACT OF THE STUDY: This innovative approach using a cheap agro-waste to control the persistent soil-borne Fusarium pathogen of cherry tomato should increase soil survival rate of Trichoderma and has potential for upscaling in the field for other crops.
Genetic diversity and pathogenicity of Fusarium species associated with fruit rot disease in banana across Peninsular Malaysia

Abd Murad NB, Mohamed Nor NMI, Shohaimi S, Mohd Zainudin NAI

J Appl Microbiol, 2017 Dec;123(6):1533-1546.
PMID: 28891270 DOI: 10.1111/jam.13582

AIMS: The aims of this study were to identify the Fusarium isolates based on translation elongation factor (tef) 1α sequence, to determine the genetic diversity among isolates and species using selected microsatellite markers and to examine the pathogenicity of Fusarium isolates causing fruit rot disease of banana.
METHODS AND RESULTS: One-hundred and thirteen microfungi isolates were obtained from fruit rot infected banana in Peninsular Malaysia. However, this study was focused on the dominant number of the discovered microfungi that belongs to the genus Fusarium; 48 isolates of the microfungi have been identified belonging to 11 species of Fusarium, namely Fusarium incarnatum, Fusarium equiseti, Fusarium camptoceras, Fusarium solani, Fusarium concolor, Fusarium oxysporum, Fusarium proliferatum, Fusarium verticillioides, Fusarium sacchari, Fusarium concentricum and Fusarium fujikuroi. All Fusarium isolates were grouped into their respective clades indicating their similarities and differences in genetic diversity among isolates. Out of 48 Fusarium isolates tested, 42 isolates caused the fruit rot symptom at different levels of severity based on Disease Severity Index (DSI). The most virulent isolate was F. proliferatum B2433B with DSI of 100%.
CONCLUSIONS: All the isolated Fusarium species were successfully identified and some of them were confirmed as the causal agents of pre- and postharvest fruit rot in banana across Peninsular Malaysia.
SIGNIFICANCE AND IMPACT OF THE STUDY: Our results will provide additional information regarding new report of Fusarium species in causing banana fruit rot and in the search of potential biocontrol agent of the disease.
Fulltext First report of Aspergillus niger causing fruit rot of bilimbi in Malaysia

Mohd Zainudin NAI, Abd Murad NB, Aris A, Hussain NH

Plant Dis, 2022 Sep 19.
PMID: 36122197 DOI: 10.1094/PDIS-06-22-1291-PDN

In July 2019 to November 2021, symptoms of fruit rot of Averrhoa bilimbi (commonly known as bilimbi) fruits were observed in Serdang (3°00'05.8"N 101°42'18.4"E) and Tanjong Karang (3°25'55.3"N 101°12'55.7"E), Selangor, Malaysia. External decay showed some yellowish to brownish-red discoloration and inside the fruit there was black powdery sporulation and a brownish decay that measured between 8 - 15 mm wide. Twenty random symptomatic fruits were collected from each location. Small pieces (5 mm) of infected tissues from the fruit rot were surface sterilized for 1 min in 0.5% NaOCl, washed twice with sterile distilled water and cultured onto potato dextrose agar (PDA) and peptone pentachloronitrobenzene agar (PPA). The plates were incubated at 28 ± 1oC under 12 hours light/dark for 7 days. The fungal colonies growing from the plates were purified using hyphal tip technique (Leyronas et al. 2012). A total of 42 fungal isolates were obtained, and the morphology characteristics of six isolates were matched that of Aspergillus niger. The A. niger isolates were further identified based genus and species-specific Internal Transcribed Spacer (ITS) sequencing. Primers ITS1/ITS4, were used to amplify and subsequently sequenced the ITS1-5.8S-ITS2 region (White et al. 1990). Primer ASAP1 (5'-CAGCGAGTACATCACCTTGG-3'), ASAP2 (5'-CCATTGTTGAAAGTTTTAACTGATT-3') was used for Aspergillus species confirmation, primer ASPU (5'-ACTACCGATTGAATGGCTCG-3') / Ni1r (5'-ACGCTTTCAGACAGTGTTCG-3') for A. niger species-specific, ASPU / Af3r (5'-CATACTTTCAGAACAGCGTTCA-3') for A. fumigatus specific-specific and ASPU / Fl2r (5'-TTCACTAGATCAGACAGAGT-3') for A. flavus specific-specific (Sugita et al. 2004). In general, Aspergillus niger isolates grew rapidly on PDA and were visibly white initially then appearing black and powdery on the second day of incubation (Figure 1A). Some isolates grew rapidly (0.71-0.85 cm/day) and have a cottony appearance. The conidia were appeared brown to black, globose and rough with diameter ranging between 4.1-5.2 µm (Figure 1B). The vesicles were hyaline, globose, and brown in color with measurement of 30-75 µm in diameter with uniseriate sterigmata (Figure 1C). The conidial head was brownish black in color and split into several irregular and regular columns of conidial chains (Figure 1D-E). The conidiophores were hyaline, and brown in color. Phylogenetic trees of ITS (Figure 2A) and ASAP sequences (Figure 2B) were constructed using a Neighbor-Joining method showing isolates Aspergillus niger #11, #15, #32, #33, #41 and #42 were grouped into the same clade as A. niger (accession no. MT446087). To examine virulence of A. niger, pathogenicity tests were performed three times by inoculating an asymptomatic fruit with six isolates of A. niger (isolate #11, #15, #32, #33, #41 and #42) and a single isolate for each species of Aspergillus aculeatus, Lasiodiplodia theobromae and Penicillium gerundense. Ten fruits were inoculated by placing a mycelial disc (6 mm) (Kouame et al. 2010) from a 5-day-old culture of each fungal colony while control fruits were non-inoculated with any fungal colony (10 fruits were inoculated with a sterile agar disc and 10 were non-inoculated, respectively). After 3 days, typical symptoms of Aspergillus fruit rot were observed on A. niger inoculated fruits, whereas the control fruits remained asymptomatic (Figure 1F-P). Aspergillus niger was reisolated and reidentified based on morphological and molecular characterization from the inoculated, symptomatic fruits, thus confirming Koch's postulates. A. niger causing widespread diseases in various plant and it is a common contaminant of food. This study shows A. niger to be highly virulent on bilimbi fruits and leads to reduction of fruit quality and its production. To our knowledge, this is the first report of A. niger causing fruit rot on bilimbi and future work on its pathogenesis may provide strategies for disease control against the pathogen.
Fulltext Plant Growth-Promoting Bacteria as an Emerging Tool to Manage Bacterial Rice Pathogens

Ngalimat MS, Mohd Hata E, Zulperi D, Ismail SI, Ismail MR, Mohd Zainudin NAI, et al.

Microorganisms, 2021 Mar 26;9(4).
PMID: 33810209 DOI: 10.3390/microorganisms9040682

As a major food crop, rice (Oryza sativa) is produced and consumed by nearly 90% of the population in Asia with less than 9% produced outside Asia. Hence, reports on large scale grain losses were alarming and resulted in a heightened awareness on the importance of rice plants' health and increased interest against phytopathogens in rice. To serve this interest, this review will provide a summary on bacterial rice pathogens, which can potentially be controlled by plant growth-promoting bacteria (PGPB). Additionally, this review highlights PGPB-mediated functional traits, including biocontrol of bacterial rice pathogens and enhancement of rice plant's growth. Currently, a plethora of recent studies address the use of PGPB to combat bacterial rice pathogens in an attempt to replace existing methods of chemical fertilizers and pesticides that often lead to environmental pollutions. As a tool to combat bacterial rice pathogens, PGPB presented itself as a promising alternative in improving rice plants' health and simultaneously controlling bacterial rice pathogens in vitro and in the field/greenhouse studies. PGPB, such as Bacillus, Pseudomonas, Enterobacter, Streptomyces, are now very well-known. Applications of PGPB as bioformulations are found to be effective in improving rice productivity and provide an eco-friendly alternative to agroecosystems.
A laudable strategy to manage bacterial panicle blight disease of rice using biocontrol agents

Ngalimat MS, Mohd Hata E, Zulperi D, Ismail SI, Ismail MR, Mohd Zainudin NAI, et al.

J Basic Microbiol, 2023 Nov;63(11):1180-1195.
PMID: 37348082 DOI: 10.1002/jobm.202300182

Bacterial panicle blight (BPB) disease is a dreadful disease in rice-producing countries. Burkholderia glumae, a Gram-negative, rod-shaped, and flagellated bacterium was identified as the primary culprit for BPB disease. In 2019, the disease was reported in 18 countries, and to date, it has been spotted in 26 countries. Rice yield has been reduced by up to 75% worldwide due to this disease. Interestingly, the biocontrol strategy offers a promising alternative to manage BPB disease. This review summarizes the management status of BPB disease using biological control agents (BCA). Bacteria from the genera Bacillus, Burkholderia, Enterobacter, Pantoea, Pseudomonas, and Streptomyces have been examined as BCA under in vitro, glasshouse, and field conditions. Besides bacteria, bacteriophages have also been reported to reduce BPB pathogens under in vitro and glasshouse conditions. Here, the overview of the mechanisms of bacteria and bacteriophages in controlling BPB pathogens is addressed. The applications of BCA using various delivery methods could effectively manage BPB disease to benefit the agroecosystems and food security.