축산 폐수는 고농도의 영양염류와 중금속을 함유하고 있어, 배출될 때 수질을 악화시킨다. 기존 처리 기술과 비교하여 bioremediation은 축산 폐수 처리에 유능하다. 특히, 미세조류는 오염물질 제거에 잠재력을 가지고 있다. 본 연구에서는 Ankistrodesmus bibraianus를 이용하여 축산 폐수 내 영양염류 (질소 (N), 인 (P))와 중금속 (구리 (Cu), 아연 (Zn))의 제거 가능성을 평가하고, A. bibraianus의 최적배양조건을 확립하였다. 연구결과, 최적 배양조건은 28°C, pH 7, 광주기는 14 : 10 h로 설정되었다. N과 P의 단일 처리구 (500, 1,000, 5,000, 10,000 mg L-1)에서 제거효율은 각각 22.9~80.6%와 11.9~50.0%였다. 또한, N과 P의 복합 처리구에서 제거효율은 각각 16.4~58.3%와 7.80~49.8%였다. Cu와 Zn의 단일 처리구 (10, 30, 50 mg L-1)에서 제거효율은 각각 15.5~81.5%와 6.28~34.3%였다. 유사하게, Cu와 Zn의 복합 처리구에서 제거효율은 각각 16.7~74.5%와 5.58~27.5%였다. 또한 영양염류 (N 및 P)와 중금속 (Cu 및 Zn)의 성장 및 제거효율을 축산 폐수에 적용할 수 있음을 나타냈다. 본 연구의 결과에 따르면 A. bibraianus는 축산 폐수 내 영양염류와 중금속 제거에 이용할 수 있을 것으로 사료된다.

In this study, the effects of single and binary heavy metals toxicity on the growth and phosphorus removal ability of Bacillus sp.. known as be a phosphorus-removing microorganism, were quantitatively evaluated. Cd, Cu, Zn, Pb, Ni were used as heavy metals. As a result of analysis of variance of the half of inhibition concentration and half of effective concentration for each single heavy metal treatment group, the inhibitory effect on the growth of Bacillus sp. was Ni < P b < Z n < Cu < C d. A nd the inhibitory effect on phosphorus removal by Bacillus s p. w as N i < Pb < Z n < Cu < C d. When analyzing the correlation between growth inhibition and phosphorus removal efficiency of a single heavy metal treatment group, a negative correlation was found (R2 = 0.815), and a positive correlation was found when the correlation between IC50 and EC50 was analyzed (R2 = 0.959). In all binary heavy metal treatment groups, the interaction was an antagonistic effect when evaluated using the additive toxicity index method. This paper is considered to be basic data on the toxic effects of heavy metals when phosphorus is removed using phosphorus removal microorganisms in wastewater.

This study was initiated to isolate the microorganisms removing phosphorus (P) from domestic sewage and to investigate the effects of environmental factors on the growth and P removal of the isolated bacteria. Microorganisms isolated from the sewage were identified as Chryseobacterium sp., Stenotrophomonas maltophilia, and Bacillus licheniformis. Among them, Bacillus licheniformis was selected as the P removal microorganism. The environmental factors considered in this study included initial phosphorus concentration, temperature, pH, and carbon source. At initial P concentrations of 10, 20, and 30 mg/L, the P removal efficiencies were 100.0%, 84.0%, and 16.5%, respectively. At 20°C, 30°C, and 40°C, the P removal efficiencies were 0%, 75.8%, and 60.6%, respectively. The removal efficiencies of phosphorus according to pH were 1.6%, 91.7%, and 51.1% at pH 5, pH 7, and pH 9, respectively. Using glucose, acetate, and glucose + acetate as carbon sources yielded P removal efficiencies of 80.9%, 33.6%, and 54.1%, respectively. Therefore, the results from the study demonstrated that the P removal efficiencies of Bacillus licheniformis were the highest when the initial P concentration, temperature, pH, and carbon source were 10 mg/L, 30°C, 7, and glucose, respectively.

This study was conducted to evaluate the effect of phosphorus acid (H3PO3) addition to the horticultural bed soil on the initial growth of red pepper (Capsicum annuum L. cv.), cucumber (Cucumis sativus L. cv.), and kimchi cabbage (Brassica campestris L. ssp. pekinensis (Lour.) Rupr. cv.). The stem heights of red pepper and cucumber were 46.1% and 23.0% greater in the 50 mg/L of phosphorus acid treatment than the untreated (control). Further, the stem diameter of pepper and cucumber were 48.7% and 23.0% greater in the 50 mg/L of phosphorus acid treatment than the control. In addition, the number of kimchi cabbage leaves was 47.5% greater in the 50 mg/L of phosphorus acid treatment than the control. The dry weights of red pepper, cucumber and kimchi cabbage were 72.9%, 16.5%, and 30.4% heavier in the 50 mg/L than the control, respectively. Cations (K, Ca, and Mg) and total phosphorus (T - P) were quantitatively analyzed for these three horticultural crops. The concentrations of K, Ca, and Mg, and T - P were higher in the 50 mg/L of phosphorus acid than the control, respectively. Based on the results obtained in this study, it appears that treatment of phosphorus acid in horticultural bed soil enhanced the growth of red pepper, cucumber and Kimchi cabbage.

Bioremediation has been recognized as a suitable alternative to conventional methods of removing contaminants, and it uses fungi, bacteria and microalgae. In contrast to other organisms, microalgae are unique in that they have the ability to perform photosynthesis like plants and to utilize organic/inorganic carbon substrates, in a process called phytoremediation. Microalgae can populate a reaction site rapidly and enhance the bioremediation efficiency. In this study, Chlorella vulgaris was used to evaluate the removal potentials of the nutrients (N and P) and heavy metals (Cu and Zn) from swine wastewater. The optimum growth conditions for Chlorella vulgaris and the removal potentials of N, P, Cu, and Zn from synthetic wastewater using Chlorella vulgaris were investigated. Based on the results, the applicability of this microalga to on-site wastewater treatment was examined. Optimal growth conditions for Chlorella vulgaris were established to be 28℃, a pH of 7, and light and dark cycles of 14:10 h. As the concentrations of the nutrients were increased, the efficiencies of N and P removal efficiencies by Chlorella vulgaris were decreased in the single and binary mixed treatments of the nutrients, respectively. Further, the efficiencies of Cu and Zn removal also decreased as the heavy metals concentrations added were increased, both in the single and binary mixed treatments. In addition, the efficiency of Cu removal was higher than that of Zn removal. Our results indicate that Chlorella vulgaris could be used in treatment plants for the removal of nutrients and heavy metals from swine wastewater.

This study was conducted to establish the type and method of fertilization for no-tillage during the third year of No-tillage (NT) and Conventional-tillage (CT) practices, towards different kinds of fertilizers. In this experiment, the livestock manure showed higher in response to fertilizer effects of no-tillage. Comparing growth characteristics and yield in NT and CT. Regarding yield, there is no significant between livestock fertilizer and chemical fertilizer, but between livestock fertilizer and chemical fertilizer in conventional fertilization has significant differences. Based on the result, livestock fertilizer is effective way on the quantity of the crop. Nitrogen absorption of plant in livestock of no-tillage is more effective than conventional fertilization. In case of the phosphorus absorption and potassium absorption of plant, fertilizer effect has no significant. Nitrogen is highly absorbed in livestock fertilization of NT. Absorption of phosphorus and potassium are similar.

The present study was conducted to establish the optimal fertilization type and method for no-tillage during the first year of No-tillage (NT) and Conventional-tillage (CT) practices for soybean, using different types of fertilizers. In this experiment, the culm length and stalk diameter showed a greater response to fertilization with surface irrigation than to conventional fertilization. The fastest flowering period (July 28) occurred using chemical fertilization applied via subsurface irrigation. Comparing maturation based on growth characteristics and flowering date revealed that fertilization with subsurface irrigation was more effective for the growth of crops than other methods. Regarding yield, there was no significant difference between livestock and chemical fertilizers in subsurface irrigation, but there were significant differences between these fertilizers when using conventional fertilization methods. Based on the results, livestock fertilizer with subsurface irrigation effectively enhanced crop quantity. Nitrogen absorption of plants using subsurface irrigation was more effective than that using conventional fertilization. Regarding phosphorus absorption of plants, chemical fertilizers showed higher absorption than did livestock fertilizers for both subsurface irrigation and conventional fertilization. Unlike nitrogen, phosphorus was highly absorbed using conventional fertilization. Absorption of phosphorus and potassium were similar but phosphorus was not absorbed using livestock fertilizers applied either using subsurface irrigation or with conventional fertilization.

Common buckwheat has the sporophytic self-incompatibility mechanism and that’s why it has the ability to cross pollinate between two plants with different styles (thepin type and thrum type). The S supergene is thought to govern self-incompatibility, flower morphology and pollen size in buckwheat. Already, we have produced self-compatible buckwheat lines by an interspecific hybridization between Fagopyrum esculentum and F. homotropicum by embryo culture. The pollen size of F1 plants produced by a cross between a pin type plant and the self-compatible plant was similar to that of the self-compatible lines and segregated together with flower morphology without exception. The pollen tubes of the self-compatible plants were compatible with styles of the pin plants but incompatible with the styles of thrum plants. But, the pollen tubes of thrum flowers were compatible with the styles of self-compatible plants. Also, the pollen tubes of pin flowers were incompatible with the styles of self-compatible plants. Already, from these results, we have reported a tentative genotype for heterostyle and homostyle flower types. Homomorphism was controlled by a single allele Sh, while the pin/thrum-complex gene was governed by a single genetic locus S, with two alleles, S and s, which control Ss (thrum-type) as well as the ss (pin-type), respectively. Corresponding represents the case of a single locus S with three alleles, Sh, S and s, and the phenotypes, homomorphic, pin and thrum. It can be characterized by relationship of dominance, S>Sh>s. Using the two self-fertile lines, one is considered as the long-homostyle flowers and the other is considered as the short homostyle flowers. If the short-homostyle trait had arisen by recombination in the S supergene, its genotype would be considered to be GIs ip a/GIs ip a. The pollen tubes of the short-homostylous plant should be compatible with the styles of thrum plants. Also, the pollen tubes of short-homostylous plants should be incompatible with the style of long-homostylous plants, and the reciprocal cross also should be incompatible, because the genotype of long homostyle is gis Ip PA/gis Ip PA. Furthermore, the flower morphology of F1plants produced by the cross between cross and short homostyle flowers should be thrum or short homostyle and only short-homostylous plants should be produced by the cross between pin and short homostyle flowers. However, the compatibility or incompatibility of short homostyle flower was not clarified. So, we need to clarify the compatibility or incompatibility of the style of short homostyle flowers for the next step.

Recently, proteome analysis is becoming a powerful tool for the functional characterization of plants. Due to the availability of vast nucleotide sequence information and based on the progress achieved in sensitive and rapid protein identification by mass spectrometry, proteome approaches open up now perspectives to analyze the complex functions of model crop species at different level. In this study, we have N-terminal sequencing data for the 100 embryo and 53 seed proteins of rice separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) were collected and systematically organized for a protein sequence data-file. An attempt was made to link the embryo proteins of rice to DNA sequences for understanding their functions. One hundred proteins of the 700 spots were detected in the embryo using 2-DE gels whereas we used micro sequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were identified to be unknown functions as previously reported by Woo et al.,. In addition, twenty-four spots of protein with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available EST databases of rice at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). Also, a total of 53 proteins out of 700 protein spots separated on the 2-DE gels were analyzed by the peptide mass fingerprinting method (MALDI-TOF/MS). High-quality mass spectra suitable for peptide mass fingerprinting were obtained from 41 spots. Using the ESI-Q-TOF/MS, however, we were able to identify 53 seed proteins of rice, including 12 proteins not registered in database. The rapid expansion of DNA sequence databases to the utilization of EST now provides the whole or partial gene sequences of model organisms, and the recent advances in protein micro-characterization by mass spectrometry allow the possibility of linking these DNA sequences to the proteins in functional complexes. Proteome Database of rice is updated, and is available on the World Wide Web at http://gene64.rda.affrc.go. This work shows that the proteome analysis could be a useful strategy to link the sequence information to the functional genomics.

We improved the separation of the basic proteins from the soybean cotyledon, Glycine max L. Merr. by searching N-terminal sequences data in proteins isolated by two-dimensional electrophoresis (2-DE). After removed Hexane, proteins were extracted from cotyledon with a urea/Triton/2-mercaptoetanol solution. Using this method, the highly reproducible isoelectric focusing (IEF) can formed with polyacrylamide gels with pH 4.0-9.8. The IEF tube gels were used as the first dimension, and proteins were visualized by second-dimensional gel electrophoresis, and identify a number of soybean cotyledon proteins using mass spectrometry in the proteome analysis. These instruments of 2-DE and IEF tube gels were used 27 cm and investigate under various conditions. The total number of spots and features was obtained by PDQuest software (Bio-Rad). In this experiments performed, the IEF tube gels and instruments afforded good reproducibility in the number of PDQuest-detected spots from gel to gel while IPG offered better reproducibility in the total number of manually detected spots from gel to gel. In conclusion, we have separated of the basic 13 proteins in soybean. The glycinin subunit separations are also considered to play important roles in soybean breeding and biochemical characterization. The improved technique will be useful to dissect the genetic control of glycinin expression in soybean.

Buckwheat sprouts are a vegetable which provides health benefit with their nutritionally important substances. Buckwheat has been considered as preventive medicine in the last decade. The present study was focused on the reference maps common (Fagopyrum esculentum Möench.) and tatary(Fagopyrum tataricum Gaertn.) buckwheat leaf and stem cultured in light and dark condition. Proteins were extracted from 7-day germinated buckwheat sprout sand separated by two-dimensional electrophoresis(2-DE) with isoelectro focusing gel over pH3 to 10. A total of more than 1520 protein spots were revealed on 2-DE gel, in which 165 proteins were identified in the basis of peptide mass fingerprinting. Functional category analysis indicated that these differentially expressed proteins mainly involved in cellular process, defense responsive, energy production, metabolism, photosynthesis, DNA recombination, DNA replication, seed storage, signal transduction, stress responsive, transcription, translation, and energy transport proteins. The pattern at protein level suggested the important roles for energy and protein metabolism-related proteins in growing sprouts under dark and light condition, accompanied by the activated of the stress responsive and growth condition. The proteomic profiling of common and tatary buckwheat will give insight for understanding buckwheat physiology and application to buckwheat industry.

Buckwheat is one of the traditional crops and has become a renewed target of interest or a popular crop as a healthy foodstuff, because it is a good source of cereal protein which is rich with essential amino acids. However, what is critical to our health is that buckwheat contains proteins which cause a allergy. Buckwheat allergy resulting from ingestion is caused by the storage proteins in the grain with molecular weights ranging from 15, 22, 35, 39 and 50 kDa proteins of the inner fractions to low, and there were clear differences in the protein compositions between the inner and outer buckwheat flour fractions. A major allergenic protein of buckwheat is Fag e 1 with molecular weight 22 kDa (BW22KD). Buckwheat allergy is an immunoglobulin E (IgE)-mediated hypersensitive response capable of causing anaphylactic shock. Buckwheat seeds were dissected to endosperm and embryo. From each fraction we extracted proteins and analyzed extracts by SDS-PAGE and 2-DE. On electrophoregrams of endosperm proteins, 6 intense bands were detected. The most intense corresponded to molecular weights ranging from 54 to 65 kDa. These proteins have been reported not to be allergenic. We show here that the allergenic buckwheat seed proteins are found only among embryo proteins. No allergenic proteins were found in the buckwheat endosperm. The results presented here lead to the proposal that patients with hypersensitivity to buckwheat flour should use only fine flour from buckwheat endosperm, as this fraction contains no allergenic proteins. At present, specific protein spots will be selected and in-gel digested for MALDI-TOF-TOF/MS analysis.

An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.

The wheat-rye translocation lines have been agriculturally developed for the resistance to the biotypes of Hessian fly as a major insect pest of wheat. In order to compare the proteomic profiles between ‘Coker797’ (non-2RL), ‘Hamlet’ (2RL), and near-isogenic line (NIL) carrying 2RL, we evaluated the protein extraction and preparation methods for two-dimensional gel electrophoresis approach. The tissues such as leaves, stems, and roots from three wheat-rye lines were extracted by following trichloroacetic acid (TCA)/acetone precipitation. In a preliminary proteome analysis, a commonly expressed protein in Hamlet and NIL strain was identified as methionine synthase annotated in Hordeum vulgare subsp. The present study will provide the experimental guideline for the proteomic study of other useful crop plant tissues.