DOI QR코드

DOI QR Code

Expression Patterns of Enzymes in Different Tissues of Oil Seed Rape (Brassica napus L.) Seedling

유료용 유채 유식물의 조직내 효소의 발현 패턴

  • Song, Yong-Su (Environment-Friendly Agriculture Research Center (EFARC), Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Seo, Dong-Jun (Environment-Friendly Agriculture Research Center (EFARC), Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Lee, Bok-Rye (Department of Animal Science, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Jung, Woo-Jin (Environment-Friendly Agriculture Research Center (EFARC), Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, Chonnam National University)
  • 송용수 (전남대학교 응용생물공학부) ;
  • 서동준 (전남대학교 응용생물공학부) ;
  • 이복례 (전남대학교 동물자원학부) ;
  • 정우진 (전남대학교 응용생물공학부)
  • Published : 2009.06.30

Abstract

To investigate expression patterns of chitinase, ${\beta}$-1,3-glucanase and peroxidase involved in biological control of phytopathogens, three oil seed rapes (Capitol, Pollen and Saturnin) were used. Activities of the enzymes in old leaves were $9.7{\sim}11.8$ unit/mg protein in chitinase, $11.1{\sim}17.3$ unit/mg protein in ${\beta}$-1,3-glucanase and $0.6{\sim}1.7$ unit/mg protein in peroxidase. Activities of the enzymes in roots were $39.2{\sim}49.0$ unit/mg protein in chitinase, $49.9{\sim}62.0$ unit/mg protein in ${\beta}$-1,3-glucanase and $2.4{\sim}3.8$ unit/mg protein in peroxidase. Chitinase and ${\beta}$-1,3-glucanase activity were the highest level in Saturnin leaves and in Capitol roots while activities of those were the lowest level in Capitol leaves. Also, chitinase and ${\beta}$-1,3-glucanase and peroxidase activity were the lowest level in Saturnin roots. Active bands of chitinase isoform in leaves (73, 51, 40, 34, and 29 kDa) and in roots (100, 57 34, and 29 kDa) tissues showed in the SDS-PAGE gel. Active bands of ${\beta}$-1,3-glucanase isoform in leaves and roots (75 and 55 kDa) tissues showed on the SDS-PAGE gel. Active staining of peroxidase showed the strongest level in leaves and roots of Pollen. Active bands of peroxidase isoform in leaves (122, 114, and 93 kDa) and in roots (135, 122, 114, and 93 kDa) tissues showed on the Native-PAGE gel. These results indicated that establishment of expression pattern of enzymes in rape tissues could play as an important role with respect to resistance of plant pathogens in rape.

식물병의 생물학적 방제에 관련한 chitinase, ${\beta}$-1,3-glucanase, peroxidase의 발현 패턴을 살펴보기 위하여 3 품종(Capitol, Pollen 및 Saturnin)의 유로용 유채를 조사하였다. 유채 old leaf에서 병발생관련 단백질의 활성 중에서 chitinase의 경우 단백질 mg당 9.7${\sim}$11.8 unit, ${\beta}$-1,3-glucanase의 경우 단백질 mg당 11.1${\sim}$17.3 unit, peroxidase의 경우 단백질 mg당 0.6${\sim}$1.7 unit를 나타내었다. 유채 뿌리내 효소의 활성 중에서 chitinase의 경우 단백질 mg당 39.2${\sim}$49.0 unit, ${\beta}$-1,3-glucanase의 경우 단백질 mg당 49.9${\sim}$62.0 unit, peroxidase의 경우 단백질 mg당 2.4${\sim}$3.8 unIt를 나타내었다. Chitinase와 ${\beta}$-1,3-glucanase 활성은 Saturnin 잎과 Capitol 뿌리내에서 가장 높았고, 반면 Capitol 잎에서 가장 낮은 수준을 보였다. 또한, chitinase, ${\beta}$-1,3-glucanase 및 peroxidase 활성은 Saturnin 뿌리내에서 가장 낮은 수준을 보였다. Chitinase 동위효소가 잎(73, 51, 40, 34, 29 kDa)과 뿌리 (100, 57 34, 29 kDa)의 SDS-PAGE 겔 상에서 보였다. ${\beta}$-1,3-glucanase 동위효소가 잎과 뿌리 (75, 55 kDa)의 SDS-PAGE 겔 상에서 보였다. Peroxidase 활성염색은 Pollen의 잎과 뿌리내에서 가장 강하게 나타났다. Peroxidase 동위효소는 잎(122, 114, 93 kDa)과 뿌리(135, 122, 114, 93 kDa)의 Native-PAGE 겔 상에서 보였다. 이상의 결과로 볼 때 유채 조직내 효소 발현 패턴의 확립은 유채 생육기간 동안 식물병에 대한 저항성과 관련하여 중요한 자료가 될 것으로 사료된다.

Keywords

References

  1. Bang JK, Lee JI, Chung DH, and Kim KJ (1990) Varietal difference of seed protein content and amino acid composition in rapeseed. Kor J Crop Sci 35, 464-475
  2. Berger LR and Reynolds DM (1958) The chitinase system of a strain of Streptomyces griseus. Biochimica et Biophysica Acta 29, 522-534 https://doi.org/10.1016/0006-3002(58)90008-8
  3. Bradford MM (1976) A rapid sensitive method for quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  4. Caruso C, Chilosi G, Caporale C, Leonardi L, Bertini L, Magro P, and Buonocore V (1999) Induction of pathogenesis-related proteins in germinating wheat seeds infected with Fusarium culmorum. Plant Sci 140, 107-120 https://doi.org/10.1016/S0168-9452(98)00199-X
  5. Chance B and Maehly AC (1955) Assay of catalases and peroxidases. Method Enzymol 2, 764-775 https://doi.org/10.1016/S0076-6879(55)02300-8
  6. Chung WJ, Kim BH, and Kim TH (1993) Evolution of carbon and nitrogen concentrations in the leaves as related to dry matter and crude protein accumulation of forage rape(Brassica napus ssp. oleifera). Kor Grassl Sci 13, 58-65
  7. Daun JK (1983) Glucosinolates in canola and rapeseed their importance, composition and analysis. Can Grain Commission 1-6
  8. Ignatius SMJ, Chopra RK, and Muthukrishnam S (1994) Effects of fungal infection and wounding on the expression of chitinases and $\beta$-1,3-glucanases in near-isogenic lines of barley. Physiol Plant 90, 584-592 https://doi.org/10.1111/j.1399-3054.1994.tb08818.x
  9. Jang YS (2002) Prospect and situation of quality improvement in oilseed rape. Kor J Crop Sci 47, 175-185
  10. Jo MH and Kim DA (1988) Effects of nitrogen fertilization levels and some additives on the chemical composition and silage quality of forage rape (Brassica napus Subsp. oleifera) I. Effectd of harvesting dates and N fertilization levels on the yield and quality of forage rape. Kor Grassl Sci 8, 33-39
  11. Johnson CM (1957) Comparative chlorine requirements of different plant species. Plant Soil 8, 337-353 https://doi.org/10.1007/BF01666323
  12. Josefsson E (1967) Distribution of thioglucosides in different parts of Brassica plants. Phytochemistry 6, 1617-1627 https://doi.org/10.1016/S0031-9422(00)82893-9
  13. Josefsson E (1971) Studies of the biochemical background to differences in glucosinolate content in Brassica napus L. I. Glucosinolate content III relation to general chemical composition. Physiol Plant 24, 150-159
  14. Jung WJ, Jin YL, Kim YC, Kim KY, Park RD, and Kim TH (2004) Inoculation of Paenibacillus illinoisensis alleviates root mortality, activates of lignification-related enzymes and induction of the isozymes in pepper plants infected by Phytophthora capsici. Biol Control 30, 645-652 https://doi.org/10.1016/j.biocontrol.2004.03.006
  15. Kim CH, Kim HS, Jung YH, and Kang YJ (1992). The hydrolysis conditions of rapeseed protein by pronase. J Korean Soc Food Nutr 21, 513-518
  16. Kini KR, Vasanthi NS, Umesh-Kumar S, and Shetty HS (2000) Purification and properties of a major isoform of $\beta$-1,3-glucanase from pearl millet seedlings. Plant Sci 150, 139-145 https://doi.org/10.1016/S0168-9452(99)00176-4
  17. Kondra ZP and Downey RK (1970) Glucosinolate content of rapeseed (Brassica napus L. and B. campestris L.) meal as influenced by pod position on the plant. Crop Sci 10, 54-56 https://doi.org/10.2135/cropsci1970.0011183X001000010021x
  18. Kragh KM, Jacobsen S, Mikkelsen JD, and Nielsen KA (1993) Tissue specificity and induction of class I, II and III chitinases in barley (Hordeum bulgare). Physiol Plant 89, 490-498 https://doi.org/10.1111/j.1399-3054.1993.tb05203.x
  19. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-683 https://doi.org/10.1038/227680a0
  20. Lingappa Y, and Lockwood JL (1962) Chitin media for selective isolation and culture of Actinomycetes. Phytopathology 52, 317-323
  21. Nocrashy AS, Kiewitt M, Mangold HK, and Mukhejee KD (1975) Nutritive value of rapeseed meals and rapeseed protein isolate. Nutr Metab 19, 145 https://doi.org/10.1159/000175657
  22. Ornstein L and Davis BJ (1964) Disc electrophoresis- I Background and theory. New York Academy of Sci 121, 321-349
  23. Trudel J and Asselin A (1989) Detection of chitinase activity after polyacrylamide gel electrophoresis. Anal Biochem 178, 362-366 https://doi.org/10.1016/0003-2697(89)90653-2
  24. Trudel J and Asselin A (1998) Detection of enzymes active on vanous $\^{a}$-1,3-glucans after denaturating polyacrylamide gel electrophoresis. Electrophoresis 19, 1788-1792 https://doi.org/10.1002/elps.1150191041
  25. Yedidia I, Benhamou N, Kapulnik Y, and Chet I (2000) Induction and accumulation of PR proteins activity during early stages of root colonization by the Mycoparasite Trichoderma harzianum strain T-203. Plant Physiol Biochem 38, 863-873 https://doi.org/10.1016/S0981-9428(00)01198-0
  26. Zasada IA and Ferris H (2004) Nematode suppression with brassicaceous amendments: application based upon glucosinolate profiles. Soil Biol Biochem 36, 1017-1024 https://doi.org/10.1016/j.soilbio.2003.12.014
  27. Ziadi S, Barbedette S, Godard JF, Monot C, Le Corre D and Silue D (2001). Production of pathogenesis-related proteins in the cauliflower (Brassica oleracea var. botrytis)-downy mildew (Peronospora parasitica) pathosystem treated with acibenzolar-S- methyl. Plant Pathol 50, 579-586 https://doi.org/10.1046/j.1365-3059.2001.00609.x

Cited by

  1. Induction of Defense Response Against Rhizoctonia solani in Cucumber Plants by Endophytic Bacterium Bacillus thuringiensis GS1 vol.22, pp.3, 2009, https://doi.org/10.4014/jmb.1107.07027