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2 Journal of Plant Protection Research

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Characterization of Fusarium isolates from rice, sugarcane and maize using RFLP-IGS

100%

Hsuan H. M. , Zakaria L. , Salleh B.

Journal of Plant Protection Research

2010

tom 50

nr 4

Isolates of Fusarium from rice, sugarcane and maize were identified as F. verticillioides, F. sacchari, F. proliferatum F. subglutinans, F. fujikuroi and F. oxysporum. The species were then characterized by restriction analysis of intergenic spacer (RFLP-IGS) using AluI, Eco88I, RsaI and XhoI. Twenty-five haplotypes were identified among the isolates of Fusarium which indicated high levels of variations. UPGMA cluster analysis was conducted to cluster the isolates and to estimate the intraspecific and interspecific variability. Isolates of F. fujikuroi from rice were clustered together with isolates of F. proliferatum from rice and maize with a similarity value of 88–100%. Isolates of F. verticilliodes from maize and sugarcane were clustered together with a similarity value ranging from 92–100%, and two isolates from rice formed another cluster. Isolates of F. oxysporum from maize and rice were clustered together with a similarity value ranging from 87–95%. Isolates of F. subglutinans from rice and maize, and F. sacchari from rice and sugarcane were also clustered together with a similarity value of 77–100%. Based on RFLP-IGS analysis, variability was observed within and between species of Fusarium from rice, maize and sugarcane and the technique could be use to complement morphological characterization and to determine genetic relationships between the species.

Izolaty Fusarium z ryżu, trzciny cukrowej i kukurydzy, określono jako F. verticillioides, F. sacchari, F. proliferatum, F. subglutinans, F. fujikuroi oraz F. oxysporum. Gatunki te scharakteryzowano przy pomocy analizy restrykcyjnej międzygenowego niekodującego odcinka genu (RLFP-IGS), wykorzystując AluI, Eco881, RsaI i Hoi. Określono dwadzieścia pięć haplotypów wśród izolatów Fusarium, co wskazywało na wysoki poziom zmienności. Przeprowadzono analizę skupień w celu pogrupowania izolatów i określenia intraspecyficznej interspocyficznej zmienności. Izolaty F. fujikuroi z ryżu zgrupowano razem z izolatami F. proliferatum z ryżu i kukurydzy wykazującymi podobieństwo 88–100%. Izolaty F. verticillioides z kukurydzy i trzciny cukrowej zgrupowano w ramach wartości podobieństwa od 92 do 100%, a dwa izolaty z ryżu tworzyły inne zgrupowanie. Izolaty F. oxysporum z kukurydzy i ryżu zgrupowano również razem, z wartością podobieństwa od 87 do 95%. Izolaty F. subglutinans z ryżu i kukurydzy i F. sacchari z ryżu i trzciny cukrowej zostały też zgrupowane razem i miały wartość podobieństwa 77–100%. W oparciu o analizę RLFP–IGS obserwowano zmienność w ramach i pomiędzy gatunkami Fusarium z ryżu, kukurydzy i trzciny cukrowej. Technika RLFP–IGS mogłaby być wykorzystywana w celu uzupełnienia charakterystyki mikroskopowej i do określenia genetycznych związków pomiędzy gatunkami.

Molecular characterization and pathogenicity of Erwinia spp. associated with pineapple [Ananas comosus (L.) Merr.] and papaya (Carica papaya L.)

100%

Ramachandran K. , Manaf U. A. , Zakaria L.

Journal of Plant Protection Research

2015

tom 55

nr 4

The Erwinia species are well-known pathogens of economic importance in Malaysia causing serious damage to high-value fruit crops that include pineapple [Ananas comosus (L.) Merr.] and papaya (Carica papaya L.).The 16S rRNA sequence using eubacteria fD1 and rP2 primers, identified two bacteria species; Dickeya zeae from pineapple heart rot, and Erwinia mallotivora from papaya dieback. Phylogenetic analysis based on the neighbor-joining method indicated that all the bacterial isolates clustered in their own taxa and formed monophyletic clades. From the pathogenicity test, all isolates of D. zeae and E. mallotivora showed pathogenic reactions on their respective host plants. Genetic variability of these isolates was assessed using repetitive sequence-based PCR (rep-PCR) fingerprinting. The results indicated interspecies, and intraspecies variation in both species’ isolates. There were more polymorphic bands shown by rep-PCR fingerprints than enterobacterial repetitive intergenic consensus (ERIC) and BOX- PCRs, however both species’ isolates produced distinguishable banding patterns. Unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis indicated that all Dickeya and Erwinia isolates from the same species were grouped in the same main cluster. Similarity among the isolates ranged from 77 to 99%. Sequencing of 16S rRNA using eubacteria fD1 and rP2 primers, and rep-PCR fingerprinting revealed diversity among Dickeya and Erwinia isolates. But this method appears to be reliable for discriminating isolates from pineapple heart rot and papaya dieback.