• 한국작물학회지
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• 제43권3호
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• pp.172-178
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• 1998

Freezing-resistant plants can survive subzero temperatures by withstanding extracellular ice formation. During cold acclimation, their leaves accumulate antifreeze proteins (AFPs) that are secreted into the apoplast and have the ability to modify the normal growth of ice crystals. Three barley, two wheat and two rye cultivars were grown under two different temperature regimes (20/16$^{\circ}C$ and 5/2$^{\circ}C$, day/night). Apoplastic proteins from winter cereals were separated by SDS-PAGE and detected with antisera to AFPs from winter rye. Apoplastic proteins accumulated to much higher levels in cold-acclimated (CA) leaves compared with nonacclimated (NA) ones in winter cereals. After cold acclimation, the protein concentration of apoplastic extracts increased significantly from 0.088 $mgmL^{-1}$ to 0.448 $mgmL^{-1}$, with about 5-fold increment. Also, the apoplastic protein content per gram leaf fresh weight in CA leaves ranged from 31 $\mu\textrm{g}$ $(gFW)^{-1}$ to 120 $\mu\textrm{g}$ $(gFW)^{-1}$ with an averaged value of 77 $\mu\textrm{g}$ $(gFW)^{-1}$, and coefficients of variation of 54.9%. The CA leaves in Musketeer (a Canadian winter rye cultivar) showed the greatest AFPs and antifreeze activity followed by 'Geurumil' (a Korean winter wheat cultivar), and 'Dongbori l' (Korean facultative barley cultivar). The proteins secreted into the wheat leaf apoplast at CA condition were more numerous than those observed in winter rye, where two $\beta$-1,3-glucanase-like proteins (GLPs), two chitinase-like proteins (CLPs) and two thaumatin-like proteins (TLPs) accumulated during cold acclimation. The proteins in barley leaf apoplast at CA conditions were a little different from those in wheat leaves. The AFPs were various among and within species. More freezing-resistant cultivars had more clear and numerous bands than less freezing-resistant ones. The high determination coefficient ($R^2$ =91 %) between freezing resistance and AFPs per gram leaf fresh weight indicated that the amount of AFPs was highly related to freezing resistance in winter cereal crops.

• 한국식품영양과학회지
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• 제42권9호
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• pp.1439-1445
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• 2013

탄수화물분해효소 ${\beta}$-amylase(BA), ${\alpha}$-amylase(BAA), $cellulose+{\beta}$-glucanase(CBG)를 쌀과 함께 침지하여 쌀가루를 제조한 뒤 이 효소처리 쌀가루 100%를 이용하여 쌀쿠키를 제조해 쿠키의 품질 특성을 조사하였다. 효소처리 쌀쿠키 반죽의 밀도는 탄수화물 분해효소의 종류 및 효소처리의 유무에 따른 유의차를 나타내지 않았다. 효소처리 쌀쿠키의 퍼짐성은 비효소처리 쌀쿠키에 비해 크게 나타났으며 효소 ${\beta}$-amylase와 ${\alpha}$-amylase를 이용하여 제조한 쌀가루 쿠키를 제조했을 때 쿠키의 퍼짐성이 큰 것을 확인하였다. 효소처리 쌀쿠키 BA, BAA, CBG의 수분함량은 3.20~3.90%로 비효소처리 쌀쿠키 비해 유의적으로 낮은 수분함량을 나타내었다(P<0.05). 쌀쿠키의 색도 측정결과, 효소처리 쌀쿠키의 L값은 비효소처리 쌀쿠키에 비해 유의적으로 낮은 값을 나타냈으며, a와 b값은 효소처리 쌀쿠키가 비효소처리 쌀쿠키에 비해 유의적으로 낮은 값을 나타내었다(P<0.05). 쌀쿠키의 경도는 효소처리 쌀쿠키가 비효소처리 쌀쿠키에 비해 높은 경도를 나타내었으나 효소의 종류에 따른 차이를 나타내지 않았다. 관능적 특성으로 쌀쿠키의 특성강도 평가에서 쿠키의 고소한 향과 균열의 정도는 효소의 유무 및 효소의 종류에 따른 차이가 나타나지 않았다. 갈색의 정도와 쿠키의 고소한 맛의 강도는 비효소처리 쌀쿠키에 비해 효소처리 쌀쿠키에서 높게 나타났으며 효소 ${\alpha}$-amylase와 ${\alpha}$-amylase로 처리한 쌀가루로 제조한 쌀쿠키(BA)가 갈색의 정도와 쿠키의 고소한 맛이 적당한 강도를 나타내었다. 경도는 효소 처리 쌀쿠키의 강도가 유의적으로 높았으며 효소의 종류에 따른 유의적인 차이는 나타내지 않아 기계적 경도와 관능적인 측면의 경도가 유사한 결과를 나타내었다. 바삭함은 경도의 강도 평가와 유사한 경향을 나타내었다. 기호도 검사 결과에서, 비효소처리 쌀쿠키는 향, 외관, 맛, 조직감 및 전반적인 기호도에서 유의적으로 낮은 기호도를 나타내었다(P<0.05). 효소 ${\alpha}$-amylase를 이용한 쌀쿠키 BAA는 향, 맛 조직감에서 높은 점수를 받아 전반적인 기호도에서 높게 나타나 효소 ${\alpha}$-amylase를 0.1%의 농도로 처리하여 제조한 쌀가루로 쿠키를 제조할 때 일반 쌀쿠키에 비해 향, 맛, 조직감에서도 좋은 기호도를 나타내는 것으로 나타나 쿠키의 소재로 적합할 것으로 사료된다.

• The Plant Pathology Journal
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• 제30권4호
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• pp.355-366
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• 2014

In this study, resistance responses were investigated during the interaction of Botrytis fabae with two faba bean cultivars expressing different levels of resistance against this pathogen, Nubaria (resistant) and Giza 40 (susceptible). Disease severity was assessed on leaves using a rating scale from 1 to 9. Accumulation levels of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase) were measured in leaf tissues at different times of infection. The expression profiles of two pathogenesis-related proteins (PRPs) encoded by the genes PR-1 and ${\beta}$-1,3-glucanase were also investigated using reverse transcription RT-PCR analysis. The accumulation of these defense responses was induced significantly in both cultivars upon infection with B. fabae compared with un-inoculated controls. The resistant cultivar showed weaker necrotic symptom expression, less ROS accumulation, a lower rate of lipid peroxidation and higher activity of the enzymatic ROS scavenging system compared with susceptible cultivar. Interestingly, ROS accumulated rapidly in the resistant leaf tissues and peaked during the early stages of infection, whereas accumulation was stronger and more intense in the susceptible tissues in later stages. Moreover, the response of the resistant cultivar to infection was earlier and stronger, exhibiting high transcript accumulation of the PR genes. These results indicated that the induction of oxidant/antioxidant responses and the accumulation of PRPs are part of the faba bean defense mechanism against the necrotrophic fungus B. fabae with a different intensity and timing of induction, depending on the resistance levels.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1542-1550
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• 2016

This is the first report that paromomycin, an antibiotic derived from Streptomyces sp. AG-P 1441 (AG-P 1441), controlled Phytophthora blight and soft rot diseases caused by Phytophthora capsici and Pectobacterium carotovorum, respectively, in chili pepper (Capsicum annum L.). Chili pepper plants treated with paromomycin by foliar spray or soil drenching 7 days prior to inoculation with P. capsici zoospores showed significant (p < 0.05) reduction in disease severity (%) when compared with untreated control plants. The disease severity of Phytophthora blight was recorded as 8% and 50% for foliar spray and soil drench, respectively, at 1.0 ppm of paromomycin, compared with untreated control, where disease severity was 83% and 100% by foliar spray and soil drench, respectively. A greater reduction of soft rot lesion areas per leaf disk was observed in treated plants using paromomycin (1.0 μg/ml) by infiltration or soil drench in comparison with untreated control plants. Paromomycin treatment did not negatively affect the growth of chili pepper. Furthermore, the treatment slightly promoted growth; this growth was supported by increased chlorophyll content in paromomycin-treated chili pepper plants. Additionally, paromomycin likely induced resistance as confirmed by the expression of pathogenesis-related (PR) genes: PR-1, β-1,3-glucanase, chitinase, PR-4, peroxidase, and PR-10, which enhanced plant defense against P. capsici in chili pepper. This finding indicates that AG-P 1441 plays a role in pathogen resistance upon the activation of defense genes, by secretion of the plant resistance elicitor, paromomycin.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1073-1079
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• 2005

To investigate plant protection, pathogenesis-related (PR) proteins and plant defense enzymes related to cell wall lignification were studied in pepper plants inoculated with antagonistic Bacillus subtilis HJ927 and pathogenic strain Phytophthora capsici. Phytophthora blight disease was reduced by $53\%$ in pepper roots when preinoculated with B. subtilis HJ927 against P. capsici. The activities of PR proteins (chitinase and ${\beta}$-1,3,-glucanase) and defense-related enzymes (peroxidase, polyphenoloxidase, and phenylalanine ammonia lyase) decreased in roots of B. subtilis+P capsid-treated plants, but increased in leaves with time. The decrease and increase were much greater in P. capsici-treated plants than in B. subtilis HJ927+P capsici-treated plants, although P. capsici-treated plants had more severe damage. Therefore, changes of enzyme activities do not seem to be directly related to plant protection. We suggest that the change of these enzymes in pathogen-treated plants may be related to plant response rather than to resistance against pathogen attacks.

• Asian-Australasian Journal of Animal Sciences
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• 제11권3호
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• pp.249-254
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• 1998

Two experiments were conducted to evaluate the effects of processing of barley on the growth performance and ileal and fecal digestibility of growing pigs. In Exp. 1, a total of 20 cannulated pigs (10.80 kg BW) were allotted to four treatments. Treatments were coarse ground barley as a control (CON), finely ground barley (FINE), extruded barley (EXT) and enzyme supplemented coarse ground barley (ENZ). In Exp. 2, a total of 100 growing pigs (36.50 kg BW) were allocated to the same treatments in completely randomized block design based on sex and body weight. In the first trial, pigs fed extruded barley showed significantly higher crude protein digestibility over pigs fed finely ground barley (p < 0.05). Pigs fed finely ground barley generally showed lower nutrients digestibility. Extrusion and ${\beta}$-glucanase supplementation showed a trend to improve nutrients digestibility. However, fine grinding rather reduced nutrients digestibility. The similar trend was found in the digestibility of essential amino acids. Fine grinding of barley significantly reduced amino acids digestibility. Extrusion and enzyme supplementation were found to improve amino acids digestibility of barley in growing pigs. In the growth trial, pigs fed extruded barley grew significantly faster than any other processed barley fed pigs. And extrusion of barley significantly improved feed/gain of pigs (p < 0.05). Fine grinding of barley and enzyme supplementation did not improve growth performance of pigs. In conclusion, fine grinding and enzyme supplementation does not appear to be an economical feed processing for growing pigs when barley is employed in the diets, while extrusion can be recommended as an effective feed processing technique for barley.

• Asian-Australasian Journal of Animal Sciences
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• 제12권6호
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• pp.988-1001
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• 1999

The rumen ecosystem is increasingly being recognized as a promising source of superior polysaccharide-degrading enzymes. They contain a wide array of novel enzymes at the levels of specific activities of 1,184, 1,069, 119, 390, 327 and $946{\mu}mol$ Reducing sugar release/min/mg protein for endoglucanase, xylanase, polygalactouronase, amylase, glucanase and arabinase, respectively. These enzymes are mainly located in the surface of rumen microbes. However, glycoside-degrading enzymes (e.g. glucosidase, fucosidase, xylosidase and arabinofuranosidase, etc.) are mainly located in the rumen fluid, when detected enzyme activities according to the ruminal compartments (e.g. enzymes in whole rumen contents, feed-associated enzymes, microbial cell-associated enzymes, and enzymes in the rumen fluid). Ruminal fungi are the primary contributors to high production of novel enzymes; the bacteria and protozoa also have important functions, but less central roles. The enzyme activities of bacteria, protozoa and fungi were detected 32.26, 19.21 and 47.60 mol glucose release/min/mL mediem for cellulose; 42.56, 14.96 and 64.93 mmol xylose release/min/mL medium after 48h incubation, respectively. The polysachharide-degrading enzyme activity of ruminal anaerobic fungi (e.g. Neocallimastix patriciarum and Piromyces communis, etc.) was much higher approximately 3~6 times than that of aerobic fungi (e.g. Tricoderma reesei, T. viridae and Aspergillus oryzae, etc.) used widely in industrial process. Therefore, the rumen ecosystem could be a growing source of novel enzymes having a tremendous potential for industrial applications.

• The Plant Pathology Journal
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• 제31권1호
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• pp.50-60
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• 2015

The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

• 미생물학회지
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• 제25권1호
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• pp.9-16
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• 1987

분리한 살리실산 자화세균 중 플라스미드블 갖는 세 균주를 션별하였다. 세 균주, KU801(pKUs, pKUS) , KUS03(pKU6 pKU9), KUS06(pKU7, pKU10)는 각각 두 개씩의 플라스미드플 가지고 있음이 전기갱동에 의해 밝혀졌고, Pseudomoηas putida로 동정되었다. 세 문주들은 모두 암피실린, 터l트라사이클린, 클로람페니콜등의 항생제에 대하여 내성을 지니며, 조사 된 방화족과 지방족 탄화수소들 중 삼리실산과 그의 중간 대사물인 카터1콜만을 이용하있다. 큰 분자량의 플라스비드(pKUS, p pKU6, pKU7)는 마이로마이신 C로 처리하였을때 큐어되며 그 빈도는 각각 0.40%, l,67%, 0.7S% 이었다. 큐어된 균주는 상리실산을 분해하지 못하였으나, 여전히 야생균주와 동일한 항생제 내성을 가지고 있었다. 살리실산 분해에 관여하는 유전자가 그들 플라스미드에 있는 것으로 판명되었다. pKU5와 pKU6의 분자량은 103, SMd, pKU7의 분자량은 101Md으로 측정되었다. SAL 플라스미드인 pKU5, pKU6, pKU7은 접합에 의해 P.putida와 P.aeruginosa로는 전달되었으나, E. coli에서는 발현되지 아니하였다.

• The Plant Pathology Journal
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• 제21권4호
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• pp.301-309
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• 2005

Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.