Characterization of Fusarium proliferatum and Fusarium verticillioides based on species-specific gene and microsatellites analysis

Fusarium species are known to cause various diseases on plantations including fruits and vegetables. The most common Fusarium that can cause plant diseases are Fusarium proliferatum and Fusarium verticillioides. Ear rot disease on maize, wilt disease on cucurbits and fruit rot disease on tomato as well as banana are example of diseases caused by these two species. The objectives of this study were to identify F. proliferatum and F. verticillioides based on species-specific primers and polymerase chain reaction (PCR) amplification and to evaluate the genetic diversity of both species based on microsatellite markers. Fifty isolates of Fusarium species that were previously collected throughout Malaysia from different hosts were identified by using species-specific PCR amplification. Twenty-nine isolates were identified as F. proliferatum and 21 isolates were identified as F. verticillioides based on species-specific primer. The genetic diversity of all the fungal isolates was evaluated by using microsatellite analysis with six established primers. Five out of six primers amplified polymorphic bands with the most effective primer showing high polymorphism were (AG)7C and (TCC)5 meanwhile one primer (TTTC)4 gave negative result with no band amplified. The phylogenetic tree that was constructed showing two different clades distinguished between F. proliferatum and F. verticillioides

DISTRIBUTION AND DIVERSITY OF FUSARIUM SPECIES ASSOCIATED WITH GRASSES IN TEN STATES THROUGHOUT PENINSULAR MALAYSIA

Fusarium  is one of   the  important genera associated with grasses as saprophytes, endophytes and  pathogens. A  study was  carried  out  on  distribution  and  diversity  of  Fusarium  species associated  with  two  groups  of   grasses  in  10  states  throughout  Peninsular  Malaysia  i.e. agricultural  grasses  (Oryza  sativa  and  Saccharum  officinarum)  and  non-agricultural  grasses (Axonopus compressus, Centhotheca lappacea, Chloris barbata, Crysopogon aciculatus, Cyanadon dactylon, Dactyloctenium  aegyptium, Digitaria  ciliaris,  Echinochloa  colona,  Eleusine  indica,  Eragrostis  amabilis, Eragrostis malayana, Eragrostis uniloides, Ischaemum magnum, Panicum brevifolium, Panicum millaneum, Panicum repens, Paspalum  commersonii, Paspalum  conjugatum, Paspalum orbiculare, Pennisetum purpureum, Sacciolepis  indica, Sporobolus diander and Sporobolus  indicus). A  total of  474  isolates were single-spored and  identified by morphological characteristics. F.  semitectum was  frequently  isolated (23.6%), followed by F. sacchari and F. fujikuroi with 15.4% and 14.6%, respectively. The other nine species were F. solani (10.3%), F. proliferatum (8.9%), F. oxysporum (7.4%), F. subglutinans (6.5%), F.  equiseti  (5.5%), F.  verticillioides  (3.4%), F.  compactum  (2.5%), F.  chlamydosporum  (1.1%) and F. longipes (0.8%). Based on the Shannon-Weiner Index, F. solani was the highest (H' = 2.62) isolated from grasses. Species of  Fusarium from O. sativa were widely diverse with 11 species, followed by non-agricultural grasses with nine species and S. officinarum with only six species. This  is  the  first  report on diversity of  Fusarium associated with grasses  in Malaysia.Key words: Oryza  sativa,  Saccharum  officinarum,  non-agricultural  grass,  Fusarium  species, diversity  and Graminea

Morphological and genetic variabilities of Fusarium species isolated from Kenaf

The objectives of this study were to identify the Fusarium species associated with kenaf based on morphological, pathological and genetic characteristics. Twenty isolates of Fusarium were obtained from root, leaf and stem of kenaf in Kuala Terengganu,Malaysia. All Fusarium isolates were identified, and classified into four species such as F. oxysporum, F. proliferatum, F.semitectum and F. solani. Based on pathogenicity test, F. oxysporum was only pathogenic and caused Fusarium wilt disease in kenaf based on disease severity index (DSI), plant height and dry weight of plant. For vegetative compatibility groups (VCGs) study, 217 nitrate non-utilizing (nit) mutants were generated with nit1 was the highest mutants recovered. All the species are genetically diverse reflected from the VCG multiple groups. F. oxysporum isolates were grouped into 5 VCGs,F. proliferatum in 3 VCGs and F. semitectum in 4 VCGs. Fusarium species isolated from kenaf were diverse based on morphology and vegetative compatibility, however only F. oxysporum caused Fusarium wilt disease

Distribution and Diversity of Fusarium Species Associated with Grasses in Ten States Throughout Peninsular Malaysia

Fusarium is one of the important genera associated with grasses as saprophytes, endophytes and pathogens. A study was carried out on distribution and diversity of Fusarium species associated with two groups of grasses in 10 states throughout Peninsular Malaysia i.e. agricultural grasses (Oryza sativa and Saccharum officinarum) and non-agricultural grasses (Axonopus compressus, Centhotheca lappacea, Chloris barbata, Crysopogon aciculatus, Cyanadon dactylon, Dactyloctenium aegyptium, Digitaria ciliaris, Echinochloa colona, Eleusine indica, Eragrostis amabilis, Eragrostis malayana, Eragrostis uniloides, Ischaemum magnum, Panicum brevifolium, Panicum millaneum, Panicum repens, Paspalum commersonii, Paspalum conjugatum, Paspalum orbiculare, Pennisetum purpureum, Sacciolepis indica, Sporobolus diander and Sporobolus indicus). A total of 474 isolates were single-spored and identified by morphological characteristics. F. semitectum was frequently isolated (23.6%), followed by F. sacchari and F. fujikuroi with 15.4% and 14.6%, respectively. The other nine species were F. solani (10.3%), F. proliferatum (8.9%), F. oxysporum (7.4%), F. subglutinans (6.5%), F. equiseti (5.5%), F. verticillioides (3.4%), F. compactum (2.5%), F. chlamydosporum (1.1%) and F. longipes (0.8%). Based on the Shannon-Weiner Index, F. solani was the highest (H' = 2.62) isolated from grasses. Species of Fusarium from O. sativa were widely diverse with 11 species, followed by non-agricultural grasses with nine species and S. officinarum with only six species. This is the first report on diversity of Fusarium associated with grasses in Malaysia

Synthesis, crystal structure and Hirshfeld surface analysis of ethyl 4-hydroxy-2-(4-hydroxyphenyl)-1-methyl-5-oxo-2,5-dihydro- 1H-pyrrole-3-carboxylate

Ethyl 4 hydroxy-2-(4-hydroxyphenyl)-1-methyl-5-oxo-2,5-dihydro-1H-pyrrole-3-carboxylate, C14H15NO5 (1) was synthesized via multicomponent reaction (MCR) of sodium diethyl oxalacetate, methylamine, and 4-hydroxybenzaldehyde. The structure of 1 was elucidated by using FT-IR, NMR and GCMS. These results were further confirmed by means of single crystal X-ray crystallography. The results showed that 1 was crystallized in orthorhombic space group Pca21 where a = 17.102(4), b = 9.923(2), c = 16.037(4), Å. The quantification of intermolecular interactions in the crystal structure was obtained by Hirshfeld surface analysis and showed that the H•••H contacts were the most dominant interactions

Isolation of microfungi from Malay traditional vegetables and secondary metabolites produced by fusarium species

Microfungi isolated from Malay traditional vegetables such as Centella asiatica, Cosmos caudatus, Oenanthe javanica, Persicaria odorata and Psophocarpus tetragonolobus are well diverse. A total of 40 isolates of the fungi were identified and classified into four genera such as Aspergillus, Fusarium, Penicillium and Trichoderma. Five species of Fusarium were morphologically identified as F. oxysporum, F. semitectum, F. proliferatum, F. solani and F. konzum. Three species of Aspergillus were identified as A. niger, A. fumigatus and A. flavus. The highest number of microfungi was isolated from Cosmos caudatus (12 isolates), followed by Persicaria odorata (9 isolates), Oenanthe javanica (8 isolates), Centella asiatica (6 isolates) and Psophocarpus tetragonolobus (5 isolates). Four isolates of Fusarium species were able to produce moniliformin (MON) and five isolates were able to produce fumonisin B1 (FB1). This is the first report on diversity of microfungi associated with some Malay traditional vegetables

Diversity of fusarium species associated with post-harvest fruit rot disease of tomato

Fusarium species is one of the common pathogens of post-harvest disease to cause rot on tomato and other perishable vegetable fruits. The objectives of this study were to determine the diversity of Fusarium isolated species from post-harvest diseases of tomato fruit, to identify the causal organisms by using phenotype characteristics and to verify the pathogens of Fusarium fruit of tomato based on pathogenicity test. Carnation leaf-piece agar (CLA) and potato dextrose agar (PDA) media were used for phenotype-based identification of the Fusarium isolates with emphasis for characterizations of the shapes and sizes of the macroconidia and microconidia, colony features, growth rates, conidiogenous cells and chlamydospores. A total of 180 Fusarium isolates were obtained from 13 locations throughout Selangor. Fusarium solani was most abundantly isolated (34%) followed by F. semitectum (31%) and F. oxysporum (31%), F. subglutinans (3%) while the least was F. equiseti (1%). Twenty seven isolates were tested for pathogenicity test by injecting 1 mL of the conidial suspension onto healthy tomatoes. All the tested Fusarium isolates were pathogenic on tomato with different severity levels. The non-inoculated controls showed no symptoms of fruit rot. The most virulent was F. oxysporum isolate B711T with DSI 93.75%, while the least were isolates of F. solani (B647T) and F. oxysporum (B727T) with DSI 37.5%. Majority of the isolated Fusarium species can potentially produce mycotoxins as their secondary metabolites. The potential production of mycotoxins by pathogenic isolates of Fusarium species in contaminated tomato fruits could pose health hazards when consumed

Essential oil constituents assessment and antioxidant profiles of Stevia rebaudiana Bertoni leaves from Malaysia

Since the 15th century, essential oils (EO) were having a strong interest as therapeutic agents and recently EO has been widely used in pharmaceutical, cosmetic, food and agricultural industries. The previous study showed that EOs from Stevia rebaudiana Bertoni (SrB) possess high antioxidant, anti-inflammatory, and antimicrobial properties. In this study, the chemical constituents of EO from SrB leaves which grown in Selangor, Malaysia were obtained by ethanol Soxhlet extraction. Total polyphenolic content (TPC), total flavonoid content (TFC), and chemical composition using a gas chromatography-mass selective detector (GC-MSD) analyses of Stevia rebaudiana Bertoni essential oil (SrB EO) were carried out. The TPC and TFC of SrB EO were 14.44±0.63 mg gallic acid equivalent (GAE) /g sample extract and 32.19±1.38 mg quercetin equivalent (QE)/g sample extract, respectively. Based on GC-MSD analyses, one hundred peaks were obtained and 28 compounds were identified where lupeyl acetate occupied the highest percentage with 14.25% of the chemical constituents. The antioxidant activities of SrB EO were studied using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6- sulphonic acid (ABTS), and ferric reducing antioxidant power (FRAP) assays. The IC50 of SrB EO using DPPH and ABTS were 71.36±1.57 µg/mL Trolox equivalent (TE) and 8.88×102±0.16 µg/mL TE, respectively while the FRAP activity was 9.55±0.26 mg/g TE. These results show the promising antioxidant properties of SrB EO

DISTRIBUTION AND DIVERSITY OF FUSARIUM SPECIES ASSOCIATED WITH GRASSES IN TEN STATES THROUGHOUT PENINSULAR MALAYSIA

Fusarium is one of the important genera associated with grasses as saprophytes, endophytes and pathogens. A study was carried out on distribution and diversity of Fusarium species associated with two groups of grasses in 10 states throughout Peninsular Malaysia i.e. agricultural grasses (Oryza sativa and Saccharum officinarum) and non-agricultural grasses (Axonopus compressus, Centhotheca lappacea, Chloris barbata, Crysopogon aciculatus, Cyanadon dactylon, Dactyloctenium aegyptium, Digitaria ciliaris, Echinochloa colona, Eleusine indica, Eragrostis amabilis, Eragrostis malayana, Eragrostis uniloides, Ischaemum magnum, Panicum brevifolium, Panicum millaneum, Panicum repens, Paspalum commersonii, Paspalum conjugatum, Paspalum orbiculare, Pennisetum purpureum, Sacciolepis indica, Sporobolus diander and Sporobolus indicus). A total of 474 isolates were single-spored and identified by morphological characteristics. F. semitectum was frequently isolated (23.6%), followed by F. sacchari and F. fujikuroi with 15.4% and 14.6%, respectively. The other nine species were F. solani (10.3%), F. proliferatum (8.9%), F. oxysporum (7.4%), F. subglutinans (6.5%), F. equiseti (5.5%), F. verticillioides (3.4%), F. compactum (2.5%), F. chlamydosporum (1.1%) and F. longipes (0.8%). Based on the Shannon-Weiner Index, F. solani was the highest (H' = 2.62) isolated from grasses. Species of Fusarium from O. sativa were widely diverse with 11 species, followed by non-agricultural grasses with nine species and S. officinarum with only six species. This is the first report on diversity of Fusarium associated with grasses in Malaysia

DISTRIBUTION AND DIVERSITY OF FUSARIUM SPECIES ASSOCIATED WITH GRASSES IN TEN STATES THROUGHOUT PENINSULAR MALAYSIA

Fusarium  is one of   the  important genera associated with grasses as saprophytes, endophytes and  pathogens. A  study was  carried  out  on  distribution  and  diversity  of  Fusarium  species associated  with  two  groups  of   grasses  in  10  states  throughout  Peninsular  Malaysia  i.e. agricultural  grasses  (Oryza  sativa  and  Saccharum  officinarum)  and  non-agricultural  grasses (Axonopus compressus, Centhotheca lappacea, Chloris barbata, Crysopogon aciculatus, Cyanadon dactylon, Dactyloctenium  aegyptium, Digitaria  ciliaris,  Echinochloa  colona,  Eleusine  indica,  Eragrostis  amabilis, Eragrostis malayana, Eragrostis uniloides, Ischaemum magnum, Panicum brevifolium, Panicum millaneum, Panicum repens, Paspalum  commersonii, Paspalum  conjugatum, Paspalum orbiculare, Pennisetum purpureum, Sacciolepis  indica, Sporobolus diander and Sporobolus  indicus). A  total of  474  isolates were single- spored and  identified by morphological characteristics. F.  semitectum was  frequently  isolated (23.6%), followed by F. sacchari and F. fujikuroi with 15.4% and 14.6%, respectively. The other nine species were F. solani (10.3%), F. proliferatum (8.9%), F. oxysporum (7.4%), F. subglutinans (6.5%), F.  equiseti  (5.5%), F.  verticillioides  (3.4%), F.  compactum  (2.5%), F.  chlamydosporum  (1.1%) and F. longipes (0.8%). Based on the Shannon-Weiner Index, F. solani was the highest (H' = 2.62) isolated from grasses. Species of  Fusarium from O. sativa were widely diverse with 11 species, followed by non-agricultural grasses with nine species and S. officinarum with only six species. This  is  the  first  report on diversity of  Fusarium associated with grasses  in Malaysia. Key words: Oryza  sativa,  Saccharum  officinarum,  non-agricultural  grass,  Fusarium  species, diversity  and Graminea