Aim: This study has been aimed (i) to isolate and identify diazotrophs from Korean rice varieties; (ii) to examine the long-term effect of N and compost on the population dynamics of diazotrophs and (iii) to realize the shot-term inoculation effect of these diazotrophs on rice seedlings. Methods and Results: Diazotrophic and heterotrophic bacterial numbers were enumerated by most probable number method and the isolates were identified based on morphological, physiological, biochemical and 16s rDNA sequence analysis. Long-term application of fertilizer N with compost enhanced both these numbers in rice plants and its environment. Bacteria were high in numbers when malate and azelaic acids were used as carbon source, but less when sucrose was used as a carbon substrate. The combined application promoted the association of diazotrophic bacteria like Azospirillum spp., Herbaspirillum spp., Burkholderia spp., Gluconacetobacter diazotrophicus and Pseudomonas spp. in wetland rice plants. Detection of nifD genes from different diazotrophic isolates indicated their nitrogen fixing ability. Inoculation of a representative isolate from each group onto rice seedlings of the variety IR 36 grown in test tubes indicated the positive effect of these diazotrophs on the growth of rice seedlings though the percentage of N present in the plants did not differ much. Conclusions: Application of compost with fertilizer N promoted the diazotrophic and heterotrophic bacterial numbers and their association with wetland rice and its environment. Compost application in high N fertilized fields would avert the reduction of N 2 -fixing bacterial numbers and their association was beneficial to the growth of rice plants. Significance and Impact of the Study: The inhibitory effect of high N fertilization on diazotrophic bacterial numbers could be reduced by the application of compost and this observation would encourage more usage of organic manure. This study has also thrown light on the wider geographic distribution of G. diazotrophicus with wetland rice in temperate region where sugarcane (from which this bacterium was first reported to be associating and thereon from other plant species) is not cultivated.
Mineral oil, peanut oil and soybean oil were compared with water and gum arabic for their suitability as adhesives for seed inoculation with peat inoculants. Inoculated seeds were stored at 4, 28 and 34°C, and sampled after 1, 3 and 9 days to determine the survival of rhizobia. Germination and nodulation tests were performed on the inoculated seeds. Results showed that oils were suitable adhesives for peat inoculants. Although the oils initially bound less inoculant to the seed, the number of surviving rhizobia was similar to that obtained by the gum arabic treatment after storage at 28 and 34°C for 3 and 9 days. An interesting finding of this experiment was that peanut and soybean oils were superior to gum arabic in supporting significantly higher numbers of chickpea rhizobia at 34°C. Inoculated seeds tested for germination and nodulation showed no adverse effects from the oil treatments. Oils hold good potential as adhesives for seed application in inoculation technology.
In order to obtain promising rice growth-promoting microbial strains that can be used as substitutes for chemical fertilizers, 172 bacterial strains were isolated from rice roots grown in Korean and Russian soils. Out of them, the strains KR076, KR083, KR181 and RRj228 showed plant growth-promoting activities on maize seedlings. Bacillus megaterium KR076 and Bacillus sp. KR083 showed both nitrogen-fixing and plant growth-promoting activities, while Rhizobium sp. KR181 and Pseudomonas sp. RRj228 appeared to support only plant growth-promotion, but not N 2 fixation. Especially, RRj228 showed high growth promoting activity at low concentrations. Inoculation studies with KR083 and RRj228 revealed a high affinity to the Japonica rice variety such as Junambyeo than the Korean Tongil type variety such as Arumbyeo. Both KR083 and RRj228 strains showed rhizoplane and/or endophytic colonization in Japonica and Tongil types rice when soaked with the bacterial suspension of 1.1 × 10 5 cfu ml -1 for six and twelve hours. However, the total bacterial cell numbers were higher in the roots of Japonica variety than in the Tongil type. In inoculation trials with Daesanbyeo rice variety, the seedlings inoculated with KR181 and RRj228 at the rate of 2.0 × 10 6 cfu ml -1 showed yield increment of 35% and 33% (p < 0.01), respectively, so that they contributed to the replacement of chemical fertilizer at half doses of N, P 2 O 5 , and K 2 O in pots. In Junambyeo rice seedlings, the strain RRj228, when inoculated with a cell suspension of 1.8 × 10 6 cfu ml -1, promoted 3.4% higher yield at 70% dose than at a full dose level of N 110 kg ha -1 in field. These results suggest that the rhizobacteria KR181 and RRj228 are prospective strains for enhancing rice performance.
The competition between green manure and forage crops frequently occurred at agricultural field because of soil fertility and livestock feeding selection. These experiments were carried out to evaluate the effects shoot and residue for green manure and forage production by leguminous crops and barley mixtures at paddy. Field experiments were conducted at paddy soil from 2008 to 2009. Treatments consisted of mixture and interseeding of barley and leguminous crops (hairy vetch and crimson clover). These treatments were divided into cutting height of 8 and 25 cm for using of green manure and forage at once. The residue biomass of 25 cm cutting height was higher than 8 cm and were no significantly between mixture and inter-seeding. However, residues of legume crop were significantly higher at inter-seeding than mixture. The shoot biomass of 8 cm cutting height was higher than 25 cm for forage using. The production of legume crop was high at the barley and hairy vetch seeding plot. The mixture of hairy vetch and barley showed the best biomass of shoot and residue for green manure and forage using at 25 cm of cutting height. Also this treatment could be possible to rice cultivation by no fertilization. Therefore, we suggested that 25 cm cutting of hairy vetch and barley mixture could be used for green manure and forage at the same time under rice-based cropping system.
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