초록 ▼

○ 연구결과
• 비료의 장기처리는 무처리, 퇴비 단용 처리, 화학비료 단용처리, 퇴비 + NPK화학비료 복합처리 등 4가지였다. 지속적으로 시비된 퇴비와 화학비료에 의해서 토양 인산 형태에 변화가 있었고 퇴비 + NPK화학비료 복합처리는 토양 전 인산을 상당히 증가시켰다. 토양의 유효인산은 NPK화학비료 처리구에서 증가되었다. 또한 퇴비와 NPK화학비료 복합처리는 유기 인산농도의 증가를 보였다. 그러나 유기인산과 전 인산의 비율은 퇴비 또는 NPK화학비료 단용 장기처리에 의해 감소하였다. 토양 내 고정화된 인산은 퇴비와 NP

○ 연구결과
• 비료의 장기처리는 무처리, 퇴비 단용 처리, 화학비료 단용처리, 퇴비 + NPK화학비료 복합처리 등 4가지였다. 지속적으로 시비된 퇴비와 화학비료에 의해서 토양 인산 형태에 변화가 있었고 퇴비 + NPK화학비료 복합처리는 토양 전 인산을 상당히 증가시켰다. 토양의 유효인산은 NPK화학비료 처리구에서 증가되었다. 또한 퇴비와 NPK화학비료 복합처리는 유기 인산농도의 증가를 보였다. 그러나 유기인산과 전 인산의 비율은 퇴비 또는 NPK화학비료 단용 장기처리에 의해 감소하였다. 토양 내 고정화된 인산은 퇴비와 NPK화학비료가 지속적으로 투여되는 동안에 상당히 증가되었다. 논토양에서의 인산 고정은 aluminum (Al)과 calcium(Ca) 보다도 iron(Fe)에 고정된 양이 높았다. 최고치의 Fe-P의 양은퇴비 + NPK화학비료 복합처리구에서 나타났다.
• 토양 효소와 미생물의 활동은 비료와 퇴비의 시비에 의해 영향을 받으며 생태학적 안정성 의척도로 사용될 수 있다. 앞서 동일비료 장기연용구(밀양 영남 농업시험장) 4개의 처리(무비구, 퇴비, NPK화학비료, 퇴비와 NPK화학비료)를 대상으로 토양의 효소활성을 측정하였다.
장기간 NPK화학비료와 퇴비를 시비한 것에서는 다른 처리보다 urease, dehydrogenase 그리고 acid phosphatease의 활성이 크게 증가했다. 또한 퇴비를 시비한 것에서는 NPK화학비료를 시비한 것보다 urease, dehydrogenase, acid phosphatase의 활성이 높게 나타났다. 그러나 arylsulfatase의 활성은 퇴비를 시비한 것과 화학비료를 시비한 것 사이에 큰 차이를 보이지 않았다. 가장 높은 미생물 수를 보인 것은 NPK화학비료와 퇴비를 모두 장기 시비한 처리구에서 나타났고, 퇴비의 시비는 NPK화학비료의 시비보다 높은 미생물 수를 나타내었다. 위의 결과로 생태학적 안정성은 퇴비만 시비하거나 혹은 NPK화학비료와 함께 퇴비를 시비함에 의해 유지됨을 볼 수 있었다.
• Iron phosphates와 aluminum phosphates의 용해도가 pH의 감소와 관련된다는 사실을 바탕으로 bromo cresol green(BCG)과 bromo phenol blue(BPB)를 이용하여 iron phosphates와 aluminum phosphates를 용해시키는 미생물의 선별을 위한 새로운 방법으로 개발하고자 사용하였다. BCG는 투명도와 가시도에서 BPB보다 우수함을 알 수 있었으며, 지시약을 첨가한 Reyes`s basal medium의 사용은 투명대의 형성이 지시약을 첨가한 Picovaskaya`s medium보다 오히려 좋음을 확인하였다. 이러한 결과는 BCG를 첨가한 Reyes`s basal medium이 iron phosphates와 aluminum phosphates를 가용화하는 미생물의 분리를 위한 가장 좋은 배지임을 나타내었다. 이러한 방법을 이용하여 성능이 우수한 4개의 iron phosphates 가용화균과 5개의 aluminum phosphates 가용화균을 신속, 용이하게 선발하였다.
• 논토양의 근권에서 상기의 방법으로 분리한 Penicillium Oxalicum CBPS-Tsa를 이용하여 인산가용화에 미치는 질소 및 탄소원의 효과를 검정하였다. Glucose, sucrose, mannitol, sorbitol와 같은 다양한 carbon sources와 arginine, sodium nitrate, potassium nitrate, ammonium chloride, ammonium sulphate를 이용하여 인산가용화능을 비교하였다. Mannitol, sorbitol, ariginine는 인산 가용화에 효과적이지 못하였고, sucrose는 Ca-p와 Al-P의 가용화에서 보다 나은 carbon source였으며, glucose는 Fe-P의 가용화에 효과가 좋았다. 모든 nitrogen sources는 인산가용화를 증가시켰지만, nitrates 첨가는 ammonium 첨가보다 인산가용화에 효과적이었다. 인산 가용화는 배지의 산성화를 나타내었으며 최대 pH의 감소는 glucose 처리에서 나타났고, nitrogen sources 중에서는 ammonium chloride 처리에서 큰 pH 감소를 나타내었다.
• 옥수수 생장에서 MPS Bacteria의 접종 효과를 보기위하여 온실 조건하에서 2번의 연속적인 옥수수 pot 재배를 시행하였다. 3가지 가용화균 (Pseudomonas striata, Burkholderia cepacia and serratia marcescens)을 접종하여 옥수수를 재배한 후 건물량, 인산 및 질소흡수량 등을 측정하였다. 인산가용화균을 접종한 식물은 접종하지 않은 식물과 비교하여 식물 신장, 건물량, N, P의 농도가 모두 높게 나타났다. 두 번의 연속적인 실험에서 P. striata 와 B. cepasia를 접종한 식물은 S. marcescens를 접종한 식물과 접종하지 않은 식물에 비하여 유의성 있는 식물 신장, 건물량, N, P의 농도의 증가를 나타내었다. P. striata 와 B. cepasia를 접종한 식물은 접종하지 않은 control 식물보다 14%의 건물량과 25%의 P 농도가 증가하였다.
• 인광석을 처리한 토양에서 담배작물의 인산 흡수와 생육에 인산가용화균(PSM)이 미치는 영향을 온실 조건하에서 실험하였다. 무비 토양에 인산비료로써 인광석을 처리한 토양과 처리 하지 않은 토양에 Penicillium oxalicum CBPS-3F-Tsa을 각각 접종한 다음 담배 작물을 재배하였다. 담배작물의 인산 농도는 인광석 처리에 의해 증가하였으며, 인산가용화균을 접종한 처리가 인산가용화균을 접종하지 않은 처리에 비해 현저히 높은 인산 농도를 나타내었다. 담배 작물에 의한 인산 흡수는 인광석과 인산가용화균 처리에 의해 증가되었으며, 특히 single rock phosphate처리에서 큰 차이를 나타내었다. 담배 작물의 생육은 인광석 처리, 특히, 인광석과 가용화균의 혼합 처리에 의해 현저히 증가하였다. 작물 생육에 인산가용화균이 미치는 긍정적인 영향은 식물체내의 인산 농도와 인산흡수에 밀접한 관계를 가지며, 이러한 결과는 Penicillium oxalicum CBPS-3F-Tsa가 토양 불용성 인산과 인광석을 가용화시켜 담배작물의 생육과 인산 흡수를 증진시킬 수 있다는 것을 나타낸다.
• 최근에는 농업적 환경에 쉽게 적용하고 미생물의 생존, 안정성, 효능, 저장, 운송과 관련된 문제들을 극복하기 위해서 미생물을 담체에 고정화하여 이용하고 있다. 이러한 인산가용화균의 고정화 기술을 개발하기 위하여 토양에서 분리한 인산가용화 미생물인 P. agglomerans를 alginate, agar, gelatin에 고정화시킨 후 tricalcium phosphate가 첨가된 액체 배지에서 배양후 인산가용화능을 비교하였으며 또한 alginate에 고정화된 균의 인산가용화능을 경시적으로 조사하였다. 고정화 담체로 alginate나 agar를 사용하였을 때 고정화된 P. agglomerans의 가용화 능이 gelatin을 담체로 사용하였을 때보다 우수하였다. alginate에 고정화된 P.agglomerans가 접종된 처리구에는 배양 일에 따라서 유효인산의 농도가 급격히 증가하였고 모든 처리 구에서 배지 안의 pH가 감소하였다.
• In vitro 상태에서 상기 기술로 고정화시킨 인산가용화균의 안전성 및 이용성을 측정하였다. 고정화된 Pantoea agglomerans는 원산지가 다른 인광석을 모두 효과적으로 가용화시켰으나, 가용화 양상은 인광석 원료의 원산지에 따라 약간 다른 경향을 나타내었다. 접종 후 6일에서 모로코 인광석(215.6 mg L^-1)과 튀니지 인광석(186.1 mg L^-1), 10일에서 이스라엘 인광석(60.98 mg L^-1)과 TCP(132.3 mg L^-1), 12일에서 중국의 인광석(48.8 mg L^-1)의 최대 가용 화능이 관찰되어졌다.
고정화된 박테리아 bead의 저장기간을 조사하기 위하여 6개월 동안 두 개의 다른 온도(4℃, room temp.)에서 저장하면서 주기적으로 활성을 측정하였다. 4℃와 실내온도에서 저장된 beads는 인산가용화능 뿐만 아니라 미생물의 개체수의 유지에 있어 큰 차이를 나타내지 않았다.
• 인광석 및 PSM의 복합처리효과를 규명하기 위하여 충북지역에서 분리한 P. agglomerans을 벼(Oryza sativa L.)에 접종하여 인산흡수 및 생육효과를 규명하기 위하여 온실 실험을 수행하였다. 처리방법은 무처리구와 6개의 처리구로 1) 종자에 세균을 접종시킨 것, 2) free cell 접종과 3) bateria가 고정된 beads의 접종방법 등 3가지 방법과 pot에 따라 인광석 처리량을 1g과 2.5g을 넣어 재배하였다. 식물 성장과 인산흡수는 인산가용화균이 접종된 처리구에서 유의성있게 증대하였다. 인광석 1g을 넣은 것과 고정된 beads를 접종한 방법이 벼 식물의 성장과 인산의 흡수에 다른 처리구보다 효과가 큰 것을 알 수 있었다. P. agglomerans 접종결과 pot에 존재하는 유효인산의 농도가 증가됨을 알 수 있었다.

Abstract ▼

· External phosphorus (P) fertilization in intensive cropping systems often exceeds P demand by crops, which leads to P accumulation in soils. Levels of different pools of soil P have been affected not only by soil properties and climatic condition but also by rate and type of P applied. This experi

· External phosphorus (P) fertilization in intensive cropping systems often exceeds P demand by crops, which leads to P accumulation in soils. Levels of different pools of soil P have been affected not only by soil properties and climatic condition but also by rate and type of P applied. This experiment was conducted to investigate the long-term applications of compost and chemical fertilizer on soil phosphorus status in paddy cropping system after addition of compost and chemical fertilizers for 34 years in rice monoculture production at National Youngnam Agricultural Experiment Station, Miryang, Korea. Four different treatments of soil amendments were selected in this study: control, compost application, NPK (nitrogen?phosphorus?potassium) fertilizer application, and compost plus NPK fertilizer application. Phosphorus status varied with the long-term applications of compost and fertilizers, and the compost plus NPK fertilizer treatment significantly increased total P in soil. Available P was increased in the treatments that received chemical fertilizers. Applications of compost and chemical fertilizers increased organic P fraction but the ratio of organic P to total P declined with application of compost or chemical fertilizers. Phosphorus-fixation was significantly increased due to the long-term application of compost and chemical fertilizers. The P fixation was highest with iron (Fe) than with aluminum (Al) and calcium (Ca) in the paddy soil. The highest Fe-P content occurred in the compost plus NPK fertilizer treatment. These results represented that the higher level of P remaining in the soil is accumulated by long-term annual application of compost and chemical fertilizers than by that of chemical fertilizer, and P accumulation might be a gradual saturation of the P-sorption capacity.
· Soil enzymes and microbial activities are affected by fertilizer and compost applications and can be used as sensitive indicators of ecological stability. Microbial population and soil enzymes viz., dehydrogenase, urease, acid phosphatase and sulfatase were determined in the long-term fertilizer and compost applied paddy soil. Soil samples were collected from the four treatments (control, compost, NPK and compost+NPK). Long-term NPK+compost application significantly increased activities of urease, dehydrogenase, acid phosphatase and arylsulfatase than all other treatments. The compost application enhanced activities of urease, dehydrogenase and acid phosphatase than the NPK application. However, arylsulfatase activity was not significantly different between compost and fertilizer application. The highest microbial population was recorded in the NPK+compost treatment. The compost application also resulted in higher microbial population than the NPK application. The above results indicate that ecological stability could be maintained by application of compost alone or with NPK.
· Two pH indicator dyes, bromo cresol green (BCG) and bromo phenol blue (BPB) were used to develop a new method for the isolation of microorganisms that solubilize iron and aluminum phosphates based on the fact that the solubilization of iron and aluminum phosphates was associated with the decrease of the pH. Bromo cresol green was found to be superior to bromo phenol blue in terms of clarity and visibility. The use of Reyes's basal medium plus dye was found to be preferable to that of Pikovaskaya's medium plus dye in terms of halo zone formation. These results indicated that Reyes's basal medium plus bromo cresol green is the best medium for the isolation of iron and aluminum phosphates-solubilizing microorganisms. Four efficient iron phosphate solubilizers and five efficient aluminum phosphate solubilizers were isolated on Reyes's basal medium containing
FePO₄ and AlPO₄, respectively, plus bromo cresol green.
· Phosphorus is one of the major plant growth limiting nutrients, despite being abundant in soils in both inorganic and organic forms. Phosphobioinoculants in the form of microorganisms can help in increasing the availability of accumulated phosphates for plant growth by solubilization. Penicillium oxalicum CBPS-Tsa, isolated from paddy rhizosphere, was studied for its phosphate solubilization in the present study. The influence of various carbon sources like glucose, sucrose, mannitol and sorbitol and nitrogen sources like arginine, sodium nitrate, potassium nitrate, ammonium chloride and ammonium sulphate were evaluated using liquid media with tricalcium phosphate (Ca-P), ferric phosphate (Fe-P) and aluminium phosphate (Al-P) respectively. The sucrose-sodium nitrate amendment influenced the production 824 mg L^-1 of maximum soluble phosphate from 5 g L^-1 of calcium phosphate source. Mannitol and sorbitol among the carbon sources and ariginine among nitrogen source were poor in influencing phosphate solubilization. While sucrose was good in influencing solubilization of tricalcium phosphate and aluminum phosphate, glucose fared better in solubilizing ferric phosphates. In general, mannitol and sorbitol were poor in influencing P solubilization from all the three phosphates sources. Though all the nitrogen sources influenced better P solubilization, nitrates were better than ammonium amendment. Ammonium chloride and ammonium sulfate amendments witnessed higher uptake of available phosphates by the fungus, indicating depletion of the source in Fe-P amendment. In all experiments, phosphate solubilization was accompanied by acidification of the media. Higher pH reduction was observed in glucose amendment, which was followed by other carbon source amendments. In the nitrogen sources, ammonium chloride amendment favoured greater pH reduction than other sources.
· Two successive in vitro experiments were carried out to examine the effect of MPS bacterial inoculation on growth, nitrogen and phosphorus accumulation of maize plants under green house condition in the same soil. There were four treatments, uninoculated control and three phosphate solubilizing bacterial inoculations, viz., Pseudomonas striata, Burkholderia cepacia and Serratia marcescens. The inoculated plants showed the higher plant height, total dry mass, nitrogen and phosphorus accumulation when compared to uninoculated control plants in both experiments. In the combined data analysis from two experiments, the plants inoculated with P. striata and B. cepacia were recorded significantly higher plant height, total dry mass and P accumulation when compared to S. marcescens inoculated plant and uninoculated control plants. The P. striata and B. cepacia inoculation enhanced the total dry matter accumulation by 14 % and phosphorus accumulation by 25% over the uninoculated control plants. The nitrogen and phosphorus concentration of maize plants were also increased due to MPS bacterial inoculation, however, the effect was not significant.
· The effect of phosphate solubilizing microbes (PSM) on plant P uptake and growth in rock phosphate applied soil was tested under a greenhouse condition. Tobacco plants were grown in nonsterilized soil inoculated with Penicillium oxalicum CBPS-3F-Tsa with or without rock phosphate application as P fertilizer. Phosphorus concentration in tobacco plants was increased by the application of rock phosphate, and inoculation of soil with PSM further significantly increased P concentration in tobacco plants compared with treatments noninoculated. Phosphorus uptake by tobacco plants were also increased by application of rock phosphate and PSM inoculation, and the increase was significant compared with rock phosphate treatment (P ≤0.05). Growth of tobacco plant was significantly increased in the treatments receiving rock phosphate, and combined application of rock phosphate and PSM further increased plant growth. The positive effect of PSM inoculation on plant growth was closely related to the enhancement in plant P concentration and uptake. These results suggest that Penicillium oxalicum CBPS-3F-Tsa can solubilize insoluble soil phosphates and rock phosphate, and promote growth and P uptake of tobacco plants.
· It is now widely accepted that immobilized microbial cells can overcome some of the problems associated with microbial survival, stability, efficacy, storage, transportation and ease of application in agricultural environments. Pantoea agglomerans, a phosphate solubilizing bacterium, was immobilized in alginate, agar and gelatin carriers. All the three immobilized carriers with bacterial cells of P. agglomerans were compared for solubilization of tricalcium phosphate in pure liquid cultures. While alginate beads were tested for phosphate solubilization on alternate days up to five days, agar beads and gelatin cubes were subjected for one time phosphate solubilization analysis after seven days. Both alginate and agar immobilized cells of P. agglomerans exhibited higher efficiency in increasing the solubilization of tricalcium phosphate than gelatin-immobilized cells. The culture filtrate of alginate beadinoculation treatment registered a rapid increase in soluble phosphate concentration upon incubation. A corresponding decrease in the pH of the medium was also observed in all the treatments.
· Pantoea agglomeransimmobilized in alginate solubilized four different rock phosphates efficiently under in vitro conditions. The solubilization pattern differed according to the rock phosphate source, where maximum solubilization of Morocco and Tunisia rock phosphates (215.6 and 186.1 mg L^-1) on 6 days, Israel rock phosphate (60.98 mg L^-1) and tricalcium phosphate (132.3 mg L^-1) on 10 days and China rock phosphate (48.8 mg L^-1) on 12 days after inoculation was observed. The shelf life of the immobilized bacteria immobilized beads stored in two different temperatures was studied for six months. Beads stored at both room temperature as well as cold storage (4℃) were found equally good in supporting the bacterial population as well as phosphate solubilizing activity. P. agglomerans immobilized in alginate might be exploited for large-scale biosolubilization of rock phosphates intended for fertilizer use.
· A phosphate-solubilizing bacterium, Pantoea agglomerans, was isolated from rhizosphere soils collected from Chungbuk area. A greenhouse experiment was conducted to study the effect of combined application of rock phosphate and P. agglomerans inoculation on plant growth and phosphate accumulation of rice (Oryza sativa L.). Apart from control that received no inputs, six treatments were planned as follows; 1) seed bacterization, 2) free cell inoculation and 3) bacteria immobilized beads inoculation, individually and in combination with 1 and 2.5 g of rock phosphate per pot. The results showed that plant growth and phosphate uptake were significantly enhanced as a result of bacterial inoculation. Bacterial inoculation in the form of immobilized beads and 1 g of rock phosphate was found to affect positively the rice plant growth and phosphorus accumulation than other treatments. The available phosphate concentration of the pot mixture also found improved as a result of P. agglomerans inoculation. A positive correlation was observed between the phosphate concentration in the pot mixture and phosphate accumulation in plant.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 10
• CONTENTS ... 16
• 목차 ... 17
• 제1장 연구개발과제의 개요 ... 18
• 제2장 국내의 기술 개발 현황 ... 23
• 제3장 연구개발내용 및 결과 ... 26
• 제1절 연구개발 수행 내용 ... 26
• 제2절 연구개발 수행 결과 ... 35
• 제4장 목표달성도 및 관련분야에의 기여도 ... 85
• 제5장 연구개발결과의 활용계획 ... 91
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 93
• 제7장 참고문헌 ... 95
• 끝페이지 ... 117