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Articles by K. Sijam
Total Records ( 10 ) for K. Sijam
  M. Pasebani , H. Yaakub , K. Sijam and A. R. Alimon
  Problem statement: Bacteria can perform a variety of beneficial functions, for example many lactic acid bacteria are responsible for fermentation of silage in the process of forage conservation. In the making of silage, epiphytic lactic acid bacteria are usually insufficient in numbers to promote efficient lactate fermentation. This study was conducted to identify the predominant indigenous bacteria, with emphasis on lactic acid bacteria, from Guinea grass (Panicum maximum). Approach: Two different condition of growth using nutrient and MRS agar were prepared for isolation of the bacteria. In total, 18 purified isolates were identified by BIOLOG identification system which comprised of 9 bacterial species. Standard plate count in the both conditions was considered. Results: Three bacterial species based on the first condition of growth were identified which were belonging to Flavimonas oryzihabitans, Enerobacter cloacae, Sphingomonas paucimobilis B. Lactic acid bacteria based on the second condition of growth were belonging to Weissella confusa, Weissella paramesenteroides, Leuconostoc mesenteroides ssp. dextranicum, Lactococcus lactis ssp. hordniae. Result of plate count showed that 8.3x103 CFU lactic acid bacteria are available per gram of fresh guinea grass. Conclusion: Three hetero-fermentative and one homo-fermentative lactic acid bacteria were identified which would be suggested to use as bacterial inoculants because of the insufficient amount of epiphytic lactic acid bacteria and the availability of pathogenic bacteria in the grass.
  N. Nejat , K. Sijam , S.N.A. Abdullah , G. Vadamalai and M. Dickinson

Abstract Problem statement: Molecular methods have been used to detect phytoplasma in Malayan Red Dwarf (MRD) ecotype of coconut for the first time in Malaysia. Detect phytoplasma from coconut showing yellowing symptoms by nested PCR and Classify phytoplasma associated with disease of coconut palms, based on analysis of 16S rRNA gene operon sequences and virtual RFLP.
Approach: Twenty MRD symptomatic palms were tested. Leaf spear, inflorescence and wood shavings from trunks of coconut palms showing yellowing symptoms were harvested from Serdang located in Selangor state. PCR assays and sequence analysis were carried out.
Results: Nested PCR with primer pairs R16F2n/R16R2 and fU5/rU3 resulted in amplification of products of approximately 1.2 kb and 890 bp respectively, from 8 out of 20 MRD symptomatic palms tested. Sequence analysis of the 16S rDNA PCR products determined that the phytoplasma strain associated with Coconut Yellow Decline (CYD) in MRD ecotype belongs to the ‘Candidatus Phytoplasma cynodontis’ (16SrXIV) group of phytoplasmas. In addition, nested R16F2n/R16R2 PCR products from 6 spear leaves and 2 inflorescences from MRD palms showed high sequence similarity to the 16S rRNA gene from coconut chloroplasts, with a similar size (approximately 1.3 kb) and a further 5 R16F2n/R16R2 PCR products from MRD inflorescences showed high sequence similarities to Bacillus spp. 16S rRNA gene sequences.
These results indicate that sequencing is a reliable method for the detection. Furthermore, trunk borings are the most reliable source of DNA for phytoplasma detection in coconuts using 16S rRNA gene primers, since there is less co-amplification of PCR products from other organisms when compared to spear leaves and inflorescences, nor from the spear leaves and inflorescences of MRD palms when primers fU5/rU3 were used in nested PCR.

  Y. Noor Shahida , Y. Awang , K. Sijam , M.A. Noriha and M.G.M. Satar
  Background and Objective: Papaya (Carica papaya) is a well-known tropical fruit consumed worldwide but its expansion is seriously impaired by many diseases including dieback caused by Erwinia mallotivora. In this study, development of dieback and changes in total phenol, total sugar, total protein, peroxidase activity, polyphenol oxidase activity and leaf photosynthesis following the inoculation of E. mallotivora on two papaya cultivars, Eksotika dan Eksotika II were reported. Methodology: Erwinia mallotivora infection (1×108 CFU mL–1, 50 μL injection) to the 8 week old seedlings at the 5th node from the apex caused dieback disease to occur as early as 3 days after inoculation. The inoculated plants were completely destroyed on day 11 after inoculation. Results: Both papaya cultivars had similar trend in their physiological and biochemical changes toward the infection. Total sugar in leaves of infected plants was higher than those in the non-infected plants, but the stem and the roots of infected plants contained markedly less sugar than those of healthy plants. Erwinia mallotivora infected tissues contained higher concentration of total phenol and total protein and these changes were coupled with higher activity of peroxidase and polyphenol oxidase. Conclusion: Plant experiencing dieback disease also had a lower leaf photosynthetic rate with reduced stomatal conductance.
  K. Keshavarz , K. Sijam , M.H. Zainal Abidin , H. Habibudin and E. Nazerian
  Primer pairs XOR-F/XOR-R2 was used for rapid identification and differentiation of thirty strains of Xanthomonas oryzae pv. oryzae (Xoo) were collected from rice fields in Penang, Kedah, Selangor and Melaka during the period from 2008 to 2010. Purified DNA was extracted using a modified CTAB method and was used in the PCR. Four hundred and seventy bp product was amplified from DNA of 30 strain using XOR-F/XOR-R2 primer pairs. Sequence similarities of the intergenic region in the 16S-23S rDNA in the Malaysian strains were as high as 99-100%. Cluster analysis based on the sequencing shows that the strains are grouped one main cluster and four groups. The minimum role of varietal influence on strain variability is partly due to the almost homogenous planting of two popular rice varieties in Peninsular Malaysia during the period of the study. On the other hand, phylogenetic analysis by using intergenic region 16S-23S divided the strains into one main cluster and four groups. The first group is represented by isolates collected from Penang and the second groups from Selangor. The third and fourth groups represented strains collected from Melaka and Kedah, respectively. The present study confirmed that direct DNA extraction from infected rice tissue by using CTAB method, followed by PCR effective methods for the identification of Xoo. Further more the results indicated that strains differentiation may be affected by the geographical areas.
  M.M. Tahat , K. Sijam and R. Othman
  An in vitro experiment was conducted to study the effect of different plant root exudates on germination of Glomus mosseae and the growth of bacterial wilt Ralstonia solanacearum. Mycorrhizal spore germination increased when the volume of Mycorrhizal Tomato Root Exudates (MTRE) increased and in contrast, a negative relationship was recorded when the volume of Non-Mycorrhizal Tomato Root Exudates (NMTRE) increased. Similarly, the Mycorrhizal Corn Root Exudates (MCRE) was able to increase the percentage of germinated spores as compared to the Non-Mycorrhizal Corn Root Exudates (NMCRE). The antagonistic effect between Ralstonia solanacearum and Glomus mosseae was also studied in this research. There was no inhibition effect of mycorrhizal and non-mycorrhizal tomato and corn root exudates on growth of R. solanacearum. The study indicated that Glomus mosseae spore germination could be influenced by the host plant or pH medium.
  M. Ramin , A.R. Alimon , J.M. Panandam , K. Sijam , A. Javanmard and N. Abdullah
  The digestion and Volatile Fatty Acid (VFA) production from rice straw and oil palm fronds by cellulolytic bacteria isolated from the termite Coptotermes curvignathus were investigated. The bacteria were Acinetobacter strain Raminalimon, Enterobacter aerogenes strain Razmin C, Enterobacter cloacae strain Razmin B, Bacillus cereus strain Razmin A and Chryseobacterium kwangyangense strain Cb. Acinetobacter strain Raminalimon is an aerobic bacterium, while the other species are facultative anaerobes. There were significant differences (p<0.05) among the bacteria for Dry Matter (DM) lost and acetic acid production from rice straw and Acinetobacter strain Raminalimon showed the highest activity. The facultative bacteria C. kwangyangense strain Cb (cfu mL-1 231x10-6, OD: 0.5), E. cloacae (cfu mL-1 68x10-7, OD: 0.5) and E. aerogenes (cfu mL-1 33x10-7, OD: 0.5) were used for digestion study with the rumen fluid microflora. The in vitro gas production technique was applied for the comparative study and the parameters measured were pH, gas (volume), dry matter lost, acetic acid, propionic acid and butyric acid concentrations. pH was not significantly (p<0.05) different among the five treatments. The bacterium C. kwangyangense strain Cb showed the highest activity (p<0.05) for DM lost, acetic acid, propionic acid and butyric acid production from rice straw when compared to the other bacterial activities. There was no significance (p<0.05) difference between the three bacteria for the dry matter lost of oil palm fronds but the production of Volatile Fatty Acids (VFA) was significantly (p<0.05) high in the treatment which was inoculated with C. kwangyangense strain Cb. The Gen Bank NCBI/EMBL accession numbers for the bacterial strains are EU332791, EU305608, EU305609, EU294508 and EU169201.
  R. Khakvar , K. Sijam , M.Y. Wong , S. Radu , J. Jones and K.L. Thong
  Pulse-Field Gel Electrophoresis (PFGE) and BOX-PCR patterns were used to determine the phylogenetic relationships among 32 strains of Ralstonia solanacearum collected from banana farms in West Malaysia. Eighty two percent (25 stains) were biovar 3 and the remainders were biovar 4. Cluster analysis based on PFGE and BOX-PCR fingerprinting showed that there was significant genetic variation among all strains within this species in West Malaysia. Patterns of DNA fingerprinting in both methods were significantly correlated with sampling sites and revealed the regional similarity among the strains. The PFGE patterns varied from 78 to 99% while BOX-PCR patterns showed a high degree of variation among R. solanacearum strains. Genomic fingerprinting by PFGE also revealed one that was significantly correlated to biovar type while BOX-PCR pattern analysis was unable to differentiate various biovars. This study clearly showed that R. solanacearum strains were phylogenetically similar within a region but diverse between regions despite biovar designation.
  M. Masyahit , K. Sijam , Y. Awang and M.G.M. Satar
  Knowing the unfavorable environment for the growth of a pathogen can be utilized as the basic information in developing appropriate strategies to prevent disease occurrence on dragon fruit. Several environmental factors including temperature, pH and salinity, as well as biotic factor including three antagonistic bacteria species, namely Bukholderia cepacia, B. multivorans and Pseudomonas aeruginosa against Bipolaris sp., Colletotrichum gloeosporioides, Botryosphaeria sp. and Monilinia sp., were investigated. Mycelial growth of all tested fungi was constantly inhibited by a temperature of 35°C, while a temperature of 25°C was quite suitable for their growth. A temperature of 30°C was favorable for the growth of Colletotrichum gloeosporioides. Under different pH condition, the growth of tested fungi was mostly inhibited by extreme pH of 4 and 10. The salinity assay showed that Monilinia sp. was not affected by all treatments among tested fungi. Only concentration 100 ppm could reduce the growth of Bipolaris sp., though its inhibition statistically affected on 4 and 6 Days after Incubation (DAI). Meanwhile, the in vitro examination of antagonistic bacteria resulted in Bukholderia multivorans which was highly effective in inhibiting the growth of examined fungi, except Monilinia sp., which was more significantly influenced by B. multivorans and B. cepacia. The proper combination of environmental modification may be useful for the growth of crop in the field as well as the storage life of the fruit at postharvest preservation.
  M. Ramin , A.R. Alimon , K. Sijam and N. Abdullah
  Bacterial strains isolated from the gut of the local termite Coptotermes curvignathus were inoculated into a buffered medium containing minerals and Whatman filter paper as the sole carbon source to observe the ability of the bacteria to digest solid substrate. The bacteria were Bacillus cereus strain Razmin A, Enterobacter aerogenes strain Razmin B, Enterobacter cloacae strain Razmin C, Acinetobacter strain Raminalimon and Chryseobacterium kwangyangense strain Cb. The Gen Bank NCBI/EMBL accession numbers for the bacterial strains were EU294508, EU305608, EU305609, EU332791 and EU169201, respectively. The ability of bacterial cultures to grow in this medium as well as to digest the filter paper was determined by visual observation after 30 days. All bacterial cultures showed growth as the medium turned cloudy and the filter paper became macerated. Chryseobacterium kwangyangense strain Cb showed yellow pigmented colonies on the filter paper. Bacillus cereus strain Razmin A showed clumps of degraded filter paper with black dots.
  M. Ramin , A.R. Alimon , N. Abdullah , J.M. Panandam and K. Sijam
  In this study the lower termite Coptotermes curvignathus (Holmgren) and higher termite Macrotermes gilvus (Hagen) were identified from different parts in the vicinity of University Putra Malaysia (UPM). We isolated three enteric bacteria from the hindgut of Coptotermes curvignathus (Holmgren). All isolates were facultative anaerobes. The isolates were identified as Enterobacter aerogenes, Enterobacter cloacae and Clavibacter agropyri (Corynebacterium) by using BIOLOG assay and Bergey’s manual. The bacteria were able to assimilate carboxymethylcellulose (CMC) and cellobiose.

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