Showing papers in "Biotropia: The Southeast Asian Journal of Tropical Biology in 2009"

ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE

Abstract: Metallothionein is an important protein for detoxifying heavy metal ions. Th is research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affi ne. Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. Th e MaMt2 cDNA was successfully isolated by PCR technique. Th e MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5α. DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. Th is cDNA is identical to mRNA of AtMt2 from Arabidopsis thaliana. It does not contain any restriction sites found in the cloning sites of pGEM-T Easy. Th e deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs.

Distribution and diversity of fusarium species associated with grasses in ten states throughout peninsular malaysia

Abstract: 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 Gramineae

Feeding ecology of tree-climbing mangrove sesarmid crabs from luzon, philippines

Abstract: Despite the large ecological study of tree-climbing mangrove sesarmid crabs in other countries, the Philippine representatives appear to have not been investigated extensively. This paper presents the feeding ecology as to dependence on mangrove trees of sesarmids in different mangrove areas of southern Luzon. This is biased on the nature of the crab habitats, arboreal climbing skills and burrowing behavior of the sesarmids: Selatium elongatum and Episesarma versicolor - exclusive mangrove tree climbers (EMTC); Sarmatium germaini - occasional mangrove tree climber (OMTC); and the non-mangrove tree-climbing (NMTC) sesarmids- Neosarmatium smithii, Perisesarma bidens and Perisesarma eumolpe. Key words: mangrove crabs, sesarmid crabs, climbing skills, burrowing skills, arboreal climbing crabs, Catanduanes, Philippines

Physic nut (jatropha curcas l.) diseases in lampung province

Abstract: Intensifi ed cultivation of physic nut (Jatropha curcas L.) could raise the importance of plant diseases. Th e objectives of this research were to diagnose diseases occurring on physic nut in Lampung Province and to determine their intensity. Field observation was conducted in four districts: South Lampung, Tanggamus, Bandarlampung, and Tulang Bawang. Disease intensity, whether expressed as disease incidence or severity, was recorded from plant samples determined by making diagonal lines across the fi eld on which fi ve observation spots were made. On each spot, fi ve plant samples were observed. Specimens were also collected and placed individually in plastic bags for laboratory observation. Th e diseases found on physic nut in Lampung Province were cercospora leaf spot, alternaria leaf spot, fusarium wilt, and bacterial wilt. In addition, leaf malformation fi rst thought to be viral disease was commonly found in many locations. Further mechanical transmission failed to produce similar symptom on tested plants and higher population of mites were found on malformed leaves than that in normal leaves. Based on the disease distribution and intensity, the most likely threatening disease in physic nut cultivation is bacterial wilt. Fusarium also caused wilt, but it was only found in one subdistrict with low incidence.

Isolation And Characterization Of Methanotrophic Bacteria From Rice Fields

Abstract: Methane is a greenhouse gas and has potency to deplete ozone layer. Rice fields are a significantly sources of atmospheric methane. Chemical fertilizers application in rice fields can increase methane emission. Methanotrophic bacteria are unique in their ability to utilize methane as a sole carbon and energy source. This research was successfully isolating and characterizing methanotrophic bacteria from rice fields in Bogor and Sukabumi, West Java, Indonesia. Determination of methane oxidation activity using Gas Chromatography showed that all isolates performed methane oxidation activity. The highest methane oxidation activity was performed by BGM 9 isolate. And DNA amplification of BGM 9 genome was performed single band of mmoX in the size of 500 bp and three bands of pmoA in the size of 1000, 750 and 500 bp respectively

Viability and plasma membrane integrity of the spotted buffalo epididymal spermatozoa after thawing with the addition of dextrose into the extender

Abstract: The objective of this study was to obtain the viability and plasma membrane integrity of the spotted buffalo epididymal sperm after addition of dextrose into Andromed® extender. Spermatozoa that have been collected from cauda epididymis were diluted with Andromed® extender as control (K) and Andromed® + 0.2% dextrose (P1) and Andromed® + 0.4% dextrose (P2) as treatments. The results showed that the quality of epididymal spermatozoa decreased during cryopreservation process. The percentage of motility after thawing in P1 (46%) and P2 (46.67%) were significantly higher (P < 0.05) compared to K (41%) as well as the percentage of live sperm in P1 (58.8%) and P2 (60%) compared to K (52.2%). The percentage of membrane integrity in P1, P2 and K were 67.4; 66.8 and 68 %, respectively. In conclusion, the addition of 0.2 and 0.4% of dextrose into Andromed® acted as an extra cellular cryoprotectant and could maintain the viability and membrane integrity of the spotted buffalo epididymal spermatozoa after thawing. Pustakawan Ludya

Exopolysaccharide production and its bioactivities of the edible pleurotus ostreatus in submerged culture

Abstract: Submerged cultures were used to study the influence of various carbon sources on the mycelial biomass and exopolysaccharide production of Pleurotus ostreatus. The antimicrobial and antioxidant activity of the exopolysaccharide were also determined. The yield of mycelial biomass of 7.06 g/l, 5.12 g/l, 4.66 g/l, and 2.96 g/l was obtained by utilization of maltose, glucose, lactose and starch as a carbon source, respectively. Furthermore, in the medium containing maltose, lactose, glucose, and starch produce 100.05 mg/l, 97.73 mg/l, 78.63 mg/l, and 70.45 mg/l of exopolysaccharide, respectively. The assay of antimicrobial and antioxidant activity showed that the exopolysaccharide is more active to inhibit the growth of B. subtilis rather than those of E. coli and S. cerevisiae. Furthermore, the exopolysaccharide was also indicated to have antioxidant activity. Key words: Pleurotus ostreatus, exopolysaccharide, antimicrobial activity, antioxidant activity

Genetic variations among aquilaria species and gyrinops versteegii using amplified fragment length polymorphism markers

Abstract: Aquilaria sp. (Thymelaeaceae) is the most valuable non wood production of forestry plant in Indonesia. It produces a fragrant resin when subjected to fungal attack and has been traded internationally known as gaharu. Knowledge of genetic diversity and relationship among species and genus is important for breeding purposes and species conservation. In this study, genetic variability of six Aquilaria species were analyzed using the AmplifiedFragment Length Polymorphism (AFLP) markers. Ten AFLP primer combinations amplified 1353 DNA fragments ranging in size from100 to 350 bp of which 1285 (95%) of them were polymorphic. Genetic similarities among Aquilaria sp. consisted of A. malaccensis, A. beccariana, A. microcarpa, and A. crassna ranged from 63.90 to 72.00 % based on Dice coefficient. The dendrogram derived by the unweighted pair group method with arithmetic mean of germplasm analysis were clustered into two main groups. Hence, a genetic variation among species is quiet high. Bootstrap values for the groups supported 70% of the cluster using a linear relationship equation of (r = 0.724, P genetic similarity for 136 pairwise comparisons of Aquilaria species. For example, A. malacensis and A. microcarpa have the highest genetic similarity (72.00%) compared with another Aquilaria species. Primer pairs E-ACG/M-CTA produced a specific fragment for A. beccariana (850 bp), A. crasna (550 bp, 180 bp, and 140 bp), A. malaccencis (1500 bp), A. microcarpa (250 bp) and Gyrinops versteegii (150 bp). Primer pairs E-ACG/M-CAA produced a specific DNA fragment only for A. beccariana (1500 bp and 100 bp). Primer pairs E-ACC/M-CAC also produced only specific fragment for A. crassna (1500 bp). Study showed the usefulness of AFLP analysis in Aquilaria sp. and its potential application for breeding and species conservation. Further, molecular diversity estimated in the present study combined with the datasets on other morphological/agronomic traits will be useful for selecting the appropriate accessions for plant improvement through conventional and molecular breeding approaches. Key words: Gaharu, Aquilaria sp., Gyrinops sp., genetic diversity, molecular marker, amplified fragment length polymorphism (AFLP)

Rapid detection of the africanized honey bee: a tool for indonesian animal quarantine

Abstract: Molecular detection methods were used to determine if Africanized Honey Bees (AHBs) are present in populations of imported Apis mellifera in Indonesia. The cytochrome b (cyt b gene) was amplified from mitochondrial DNA and digested with the BglII restriction enzyme (cytb/BglII). Two types of animal DNA extraction kits were used and found suitable for rapid preparation of DNA from A. mellifera by the Animal Quarantine facility. Results showed that all 94 colony samples from beekeepers in Java produced a 485 bp PCR product from the amplification of this gene. Two DNA fragments of 194 and 291 bp from all samples were produced after digestion with BglII. This cytb/BglII result together with the DNA sequence of cyt b showed that all collected samples of A. mellifera were the non-AHB type. Hence, this study did not detect AHB in Indonesia. Key words : Apis mellifera, molecular detection methods, DNA, cytochrome b, mitochondrial genetics

Development of antibody to ralstonia solanacearum and its application for detection of bacterial wilt

Abstract: The serological assay for the detection of bacterial wilt caused by Ralstonia solanacearum (RS) was able to provide information regarding the presence of the pathogen in plant materials. The research is was aimed to develop polyclonal antibody (PAb) for RS detection. Bacterial whole cells of RS isolates mixed with glutaraldehyde were used to immunize New Zealand female white rabbit. The titre of antibody in culture supernatant was 1: 1024. The PAb developed from a ground nut RS isolates reacted with infected plant samples from various locations. It was able to detect RS antigen of crude extract and pure cultures from tomato and potato plant samples 4-5 using dot blot ELISA; however, the minimum detectable concentration of RS antigen was 10 cells/ml. The PAb obtained in this study is sensitive enough to detect RS isolates in routine serological assay. Key words: Bacterial wilt, Ralstonia solanacearum, antibody, dot blot ELISA.

Cloning of a gene encoding protein belonging to abc transporter involved in bacterial magnetic particle synthesis in magnetospirillum magneticum AMB-1

Abstract: Magnetospirillum magneticum AMB-1 synthesizes intracellular magnetic particles, magnetite (Fe3 O4 ), enveloped by membrane called magnetosome under micro-aerobic conditions. Initial study of random transposon-based mutagenesis generated 62 nonmagnetic mutants of AMB-1 in a mini-Tn5 library. In order to identify a gene involved in bacterial magnetic particle (BMP) synthesis in the magnetic bacterium M. magneticum AMB-1, a nonmagnetic mutant from the library designated as NMA38-4, was analyzed. h e amino acid sequence deduced from the gene directly interrupted by transposon, ORF4 (1482 bp), showed homology to ATP binding cassette (ABC) transporter of Mesorhizobium loti with 62 % identity and 74 % similarity. It was strongly indicated by the occurrence of putative consensus sequence of ATP-binding motifs (ATP- binding protein). h e ORF4 was subsequently cloned in pET-15b and the recombinant ORF4- Histag fusion protein was heterologously expressed in Escherichia coli BL21 (DE3) pLysS. A 55 kDa protein corresponding to the ORF4-Histag fusion protein was obtained after purifi cation using Ni-NTA column. h is is the fi rst report describing a gene cluster containing gene encoding protein belonging to ABC transporter organized in an operon which is involved in BMP synthesis. Key words: Magnetospirillum magneticum AMB-1, Bacterial Magnetic Particle (BMP), ATP Binding Cassette (ABC) Transporter, transposon mutagenesis.