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Edi Husen
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SCREENING OF SOIL BACTERIA FOR PLANT GROWTH PROMOTION ACTIVITIES IN VITRO Edi Husen
Indonesian Journal of Agricultural Science Vol 4, No 1 (2003): April 2003
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v4n1.2003.p27-31

Abstract

Fourteen isolates of soil bacteria, including two known plant growth promoting rhizobacteria (PGPR) strains, Azotobacter vinelandii Mac 259 and Bacillus cereus UW 85, were tested in vitro. Parameters assessed were indoleacetic acid (IAA) production, phosphate solubilization, dinitrogen fixation, and siderophore (Fe-III chelating agent) production. IAA production was assayed colorimetrically using ferric chlorideperchloric acid reagent. Phosphate-solubilization and siderophore production were tested qualitatively by plating the bacteria in Pikovskaya and chrome azurol S agar, respectively. The ability to fix dinitrogen was measured based on nitrogenase activity of the bacteria by gas chromatography. The results showed that twelve isolates produced IAA, ranged from 2.09 to 33.28 µmol ml-1. The ability to solubilize precipitated phosphate was positively exhibited by four isolates (BS 58, BTS, TCaR 61, and BTCaRe 65). Seven isolates including Mac 259 positively produced siderophore. None of the isolates showed nitrogenase activity. Only one isolate (TS 3) did not exhibit any of the traits tested. Isolate TCeRe 60 and reference strain Mac 259 were found to have IAA- and siderophore-producing traits. Four P-solubilizing bacteria (BS 58, BTS, TCaR 61, and BTCaRe 65) were also IAA- and siderophore-producing bacteria. Potential use of these PGPR isolates needs further test in enhancing plant growth.
THE USE OF gusA REPORTER GENE TO MONITOR THE SURVIVAL OF INTRODUCED BACTERIA IN THE SOIL Edi Husen
Indonesian Journal of Agricultural Science Vol 6, No 1 (2005): April 2005
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v6n1.2005.p32-38

Abstract

An effective marker to monitor the survival of introduced bacteria in the soil is required for further evaluation of their beneficial effects on plant growth. This study tested the use of gusA gene as a marker to trace the fate of three Gram negative bacteria in the root, rhizosphere, and soil. The study was conducted at the laboratory and greenhouse of the National Institute of Molecular Biology and Biotechnology, Philippines from January to December 2001. Isolates TCaR 61 and TCeRe 60, and Azotobacter vinelandii Mac 259 were selected as test bacteria based on their ability to produce indole-3acetic acid and solubilize precipitated phosphate, which may promote plant growth in the field. These bacteria were marked with gusA reporter gene from Escherichia coli strain S17-1(λ-pir) containing mTn5SSgusA21. The gusA (β-glucuronidase) gene from the donor (E. coli) was transferred to each bacterium (recipient) through bacterial conjugation in mating procedures using tryptone-yeast agar followed by the selection of the transconjugants (bacteria receiving gusA) in tryptone-yeast agar supplemented with double antibiotics and X-GlcA (5bromo-4chloro- 3indoxyl-β-D-glucuronic acid). The antibiotics used were rifampicin and either streptomycin or spectinomycin based on antibiotic profiles of the donor and recipients. The results showed that the insertion of gusA gene into bacterial genomes of the recipient did not impair its phenotypic traits; the growth rates of the transconjugants as well as their ability to produce indole-3acetic acid and solubilize precipitated phosphate in pure culture were similar to their wild types. All transconjugants colonized the roots of hot pepper (Capsicum annuum L.) and survived in the rhizosphere and soil until the late of vegetative growth stage. The distinct blue staining of transconjugants as the expression of gusA gene in media containing X-GlcA coupled with their resistance to rifampicin and streptomycin or spectinomycin made them easier to be recognized and evaluated.
SOYBEAN SEEDLING ROOT GROWTH PROMOTION BY 1-AMINOCYCLOPROPANE-1-CARBOXYLATE DEAMINASE-PRODUCING PSEUDOMONADS Edi Husen; Aris Tri Wahyudi; Antonius Suwanto; Rasti Saraswati
Indonesian Journal of Agricultural Science Vol 10, No 1 (2009): April 2009
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v10n1.2009.p19-25

Abstract

Pseudomonad producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase (E.C.4.1.99.4) has been known to promote plant growth by lowering ethylene biosynthesis in higher plants, which can be induced by indole-3-acetic acid (IAA) production. The objective of this study was to examine the ability of IAAproducing Pseudomonas isolated from local soil environment (rhizosphere of soybean grown in Plumbon's agricultural areain Cirebon, West Java, Indonesia) to promote soybean root growth in relation to their ACC deaminase activities. The experiments were conducted in growth room and Laboratory of Soil Biology Research, Indonesian Soil Research Institute, Bogor, from January to August 2008. Soybean seeds were inoculated by immersing the seeds for 1 hour in bacterial cell suspension containing approximately 108-109 cells ml-1. The seeds were then germinatedfor 2 days before planting in growth pouches containing sterilized distilled water. All treated and untreated seeds were grown for 7 days in growth room at 24°C with 1300 lux of light intensity for 12-hour followed by a 12-hour dark period at 22°C. ACC deaminase activity of the isolates was assayed based on their ability to grow in Dworkin-Foster’s salt minimal medium containing ammonium sulfate or ACC as a source of nitrogen. Thirteen out of 81 isolates tested significantly increased soybean root length and weight, up to 50% from untreated plants. Of 13 isolates, 11 demonstrated ACC deaminase activities. Two isolates that did not show ACC deaminase activities had lower capacity to produce IAA. The results suggest that the effectiveness of IAA producing Pseudomonas in promoting the growth of the soybean seedlings is associated with their ACC deaminase activities or they produce IAA at low levels.
Co-Authors Antonius Suwanto dan Meity S. Sinaga . Budi Tjahjono Andi Khaeruni R Aris Tri Wahyudi Rasti Saraswati