Genetic variations among the isolates of Bipolaris Maydis based on phenotypic and molecular markers

Maydis leaf blight, caused by Bipolaris maydis, is an important disease of maize crop in Khyber Pakhtunkhwa (KP) Pakistan. Fifteen isolates of the pathogen, collected across KP, were studied for variability based on phenotypic and molecular markers. Significant variability among the isolates was observed when assessed using phenotypic traits such as radial growth, spore concentration, fungicide sensitivity and virulence. The isolates were classified into six culture groups based on colour, texture and margins of the colony. Conidial morphology was also variable. These were either straight or slightly curved and light to dark brown in colour. Fungicide test showed significant variation in the degree of sensitivity against Carbendazim. Isolate Bm8 exhibited maximum radial growth on carbendazim spiked plates. Conversely, isolate Bm15 showed the lowest radial growth. Variations in virulence pattern of the isolates were evident when a susceptible maize variety Azam was inoculated with spores of B. maydis. Genetic variability amongst the isolates was also estimated by RAPD as well as sequencing of ITS region. The RAPD dendrogram grouped all the isolates into two major clusters. Average genetic distance ranged from 0.6% to 100%, indicating a diverse genetic gap among the isolates. Maximum genetic distance was found between isolates Bm9 and Bm10 as well as Bm2 and Bm8. Conversely, isolates Bm13 and Bm15 were at minimum genetic distance. Phylogenetic dendrogram based on sequencing of ITS region grouped all the isolates into a single major cluster. The clusters in both the dendrogram neither correlate to the geographical distribution nor to the morphological characteristics.

Fungal Species Causing Maize Leaf Blight in Different Agro-Ecologies in India

Foliar diseases of maize cause severe economic losses in India and around the world. The increasing severity of maize leaf blight (MLB) over the past ten years necessitates rigorous identification and characterization of MLB-causing pathogens from different maize production zones to ensure the success of resistance breeding programs and the selection of appropriate disease management strategies. Although Bipolaris maydis is the primary pathogen causing MLB in India, other related genera such as Curvularia, Drechslera, and Exserohilum, and a taxonomically distant genus, Alternaria, are known to infect maize in other countries. To investigate the diversity of pathogens associated with MLB in India, 350 symptomatic leaf samples were collected between 2016 and 2018, from 20 MLB hotspots in nine states representing six ecological zones where maize is grown in India. Twenty representative fungal isolates causing MLB symptoms were characterized based on cultural, pathogenic, and molecular variability. Internal Transcribed Spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase (GADPH) gene sequence-based phylogenies showed that the majority of isolates (13/20) were Bipolaris maydis. There were also two Curvularia papendorfii isolates, and one isolate each of Bipolaris zeicola, Curvularia siddiquii, Curvularia sporobolicola, an unknown Curvularia sp. isolate phylogenetically close to C. graminicola, and an Alternaria sp. isolate. The B. zeicola, the aforesaid four Curvularia species, and the Alternaria sp. are the first reports of these fungi causing MLB in India. Pathogenicity tests on maize plants showed that isolates identified as Curvularia spp. and Alternaria sp. generally caused more severe MLB symptoms than those identified as Bipolaris spp. The diversity of fungi causing MLB, types of lesions, and variation in disease severity by different isolates described in this study provide baseline information for further investigations on MLB disease distribution, diagnosis, and management in India.

Biosynthesis of Silver Nanoparticles by Conyza canadensis and Their Antifungal Activity against Bipolaris maydis

Silver nanoparticles were biosynthesized from Conyzacanadensis leaf extract with the help of a microwave oven. The UV-vis spectrum showed the maximum absorption at 441 nm, corresponding to the surface plasmon resonance of silver nanoparticles. Transmission electron microscope and scanning electron microscope images showed that the synthesized silver nanoparticles were spherical or near-spherical with an average diameter of 43.9 nm. X-ray diffraction demonstrated nanoparticles with a single-phase cubic structure. As-synthesized silver nanoparticles displayed prominent antifungal activity against Bipolaris maydis. The colony inhibition rate reached 88.6% when the concentration of nanosilver colloid was 100 μL·mL−1 (v/v). At such a concentration, no colony formation was observed on the solid plate. The diameter of the inhibition zone was 13.20 ± 1.12 mm. These results lay the foundation for the comprehensive control of plant pathogens using an environmentally friendly approach.

Compatibility of biocontrol agent formulas and synthetic fungicides in controlling maydis leaf blight on corn caused by Bipolaris maydis

Maydis leaf blight caused by Bipolaris maydis is one of the limiting factors in increasing maize production. The application of biological control agents (BCAs) as a biopesticide to control pests and plant diseases expected to decrease synthetic fungicide usage and its impacts. Several researches showed that the formula of Bacillus subtilis biopesticide suppress the development of several plant diseases including maydis leaf blight in maize. Nevertheless, biological control agents cannot completely replace the need for chemical fungicides in the agro-ecosystem. Therefore, this study was carried out to evaluate the effect and compatibility of several synthetic fungicides with antagonistic bacterial formulas in controlling B. maydis on maize. Five types of synthetic active ingredients used were difenoconazole, propinep, fluopicolide, metalaxyl, and dimethomorph. From the results of laboratory tests, 3 types of active ingredients with the highest inhibitory effectiveness be selected for the field test. Field treatments were arranged using a Randomized Block Design with 2 factors, factor I was biological control agents with 2 levels (T0 = without biological control agents, T1 = with biological control agents) and factor II was synthetic fungicide with different active ingredients with 4 levels (S0 = no synthetic fungicide, S1=diphenoconazole, S2=propinep, S3 = fluopicolide). Each treatment combination was repeated 4 times. The result showed that all the synthetic fungicides tested were compatible with the biological control agent. Fungicide with active ingredient of diphenoconazole was the most compatible with the BCAs showing the lowest disease severity value of 27%, while control treatment conditions without BCAs was 72%.

THÀNH PHẦN HÓA HỌC CỦA SINH KHỐI CỦA LOÀI NẤM BIPOLARIS MAYDIS

Dữ liệu thành phần hóa học của Bipolaris maydis ít được nghiên cứu. Loài nấm này được phân lập từ Kandelia candel. Ba hợp chất cis-cyclo(L-Val-L-Phe) (1), melithasterol B (2) và chrysophanol (3) đã được phân lập từ sinh khối của loài nấm Bipolaris maydis bằng nhiều phương pháp sắc kí khác nhau. Cấu trúc hóa học của các hợp chất được xác định bằng các phương pháp phổ nghiệm NMR cũng như so sánh với dữ liệu đã công bố. Ba hợp chất trên lần đầu tiên được phân lập từ chi Bipolaris.

An arabinose-induced enhancement of asexual reproduction and concomitant changes in metabolic state in the filamentous fungus Bipolaris maydis

l-Arabinose, a major constituent pentose of plant cell-wall polysaccharides, has been suggested to be a less preferred carbon source for fungi but to be a potential signalling molecule that can cause distinct genome-wide transcriptional changes in fungal cells. Here, we explore the possibility that this unique pentose influences the morphological characteristics of the phytopathogenic fungus Bipolaris maydis strain HITO7711. When grown on plate media under different sugar conditions, the mycelial dry weight of cultures on l-arabinose was as low as that with no sugar, suggesting that l-arabinose does not substantially contribute to vegetative growth. However, the intensity of conidiation on l-arabinose was comparable to or even higher than that on d-glucose and on d-xylose, in contrast to the poor conidiation under the no-sugar condition. To explore the physiological basis of the passive growth and active conidiation on l-arabinose, we next investigated cellular responses of the fungus to these sugar conditions. Transcriptional analysis of genes related to carbohydrate metabolism showed that l-arabinose stimulates carbohydrate utilization through the hexose monophosphate shunt (HMP shunt), a catabolic pathway parallel to glycolysis and which participates in the generation of the reducing agent NADPH (the reduced form of nicotinamide adenine dinucleotide phosphate). Then, the HMP shunt was impaired by disrupting the related gene BmZwf1, which encodes glucose-6-phosphate dehydrogenase in this fungus. The resulting mutants on l-arabinose showed remarkably decreased conidiation, but a conversely increased mycelial dry weight compared with the wild-type. Our study demonstrates that l-arabinose acts to enhance resource allocation to asexual reproduction in B. maydis HITO7711 at the cost of vegetative growth, and suggests that this is mediated by the concomitant stimulation of the HMP shunt.