DOI QR코드

DOI QR Code

Immunolocalization of Wound-Inducible Insoluble Acid Invertases in Pea (Pisum sativum L)

완두콩(Pisum sativum L.) 상처에서 유도되는 불용성 산성 인버타제의 면역조직화

  • Kim, Donggiun (Department of Life Science, Silla University) ;
  • Lee, Taek-Kyun (South Sea Institute, Korea Institute of Ocean Science and Technology)
  • 김동균 (신라대학교 생명과학과) ;
  • 이택견 (한국해양과학기술원 남해연구소)
  • Received : 2015.08.20
  • Accepted : 2015.09.11
  • Published : 2015.09.30

Abstract

Invertase, that hydrolyzes sucrose into glucose and fructose, plays a great role in carbohydrate reallocation between the photosynthetic source tissue and various sink tissues. Invertase also occurs in a variety of isoforms for various functions in plants. Insoluble invertases were extracted only in buffer solutions containing high concentrations of salt. Within these classes, acid invertase has an optimum activity at acidic pH (pH 4-5). Induction of insoluble acid invertase (INAC-INV) in leaf, stem, and root tissues in response to physical wounding has been investigated. To detect the localization of INAC-INV within the plant, immunolocalization has been performed. In this study, the accumulation of INAC-INV was noticeable to reach maximum levels on 72 hr after mechanical injuries. INAC-INV was induced in wounded leaves 3 times more than control leaves. Immunolocalization results showed that INAC-INV accumulated in wall appositions and intercellular spaces. INAC-INV was also localized at sieve cell walls in phloem tissues close to the site of wounding. Taken together, this study suggested that INAC-INV induction upon wounding injuries can play a role on responses to the high energy demand for wound healing process.

인버타제는 설탕을 포도당과 과당으로 가수분해하며, 광합성 조직과 다양한 수용체 조직 사이의 탄수화물 재배치에서 중요한 역할을 수행한다. 인버타제는 식물에서 다양한 기능을 수행하기 위하여 여러가지 isoforms으로 존재한다. 불용성 인버타제는 고농도의 염을 포함하는 버퍼용액에서만 추출되며, 이 종류의 인버타제 중 산성 인버타제는 산성 pH(pH 4-5)에서 최적의 활성을 갖는다. 물리적인 상처에 반응하여 잎, 줄기 및 뿌리에서 불용성 산성 인버타제(INAC-INV)가 유도되는 것이 연구되어 왔다. 식물에서 효소의 localization을 검출하기 위한 면역조직화 연구를 수행하였다. 이 연구에서 인버타제의 축적은 기계적 손상 후 72시간에 최고수준에 도달하였다. INAC-INV의 활성은 대조구 잎보다 상처받은 조직에서 3배까지 증가하였다. 면역조직화 결과는 INAC-INV가 세포벽과 세포간극에 축적되어 있음을 보여주었다. INAC-INV는 또한 상처와 가까운 사부조직의 체관세포벽에 위치하였다. 종합해 볼 때, 이 연구는 상처에 의한 INAC-INV 유도는 상처치료 과정에서 필요한 높은 에너지 요구에 대한 반응에 역할을 할 수 있음을 추측케 한다.

Keywords

References

  1. K. Koch. "Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development". Curr. Opin. Plant Biol. 7, 235-246, 2004. DOI: http://dx.doi.org/10.1016/j.pbi.2004.03.014
  2. F. Rolland, E. Baena-Gonzalez, J. Sheen. "Sugar sensing and signaling in plants: conserved and novel mechanisms". Annu. Rev. Plant Biol. 57, 675-709, 2006. DOI: http://dx.doi.org/10.1146/annurev.arplant.57. 032905.105441
  3. J. Wind, S. Smeekens, J. Hanson. "Sucrose: metabolite and signaling molecule". Phytochemistry 71, 1610-1614, 2010. DOI: http://dx.doi.org/10.1016/j.phytochem.2010. 07.007
  4. T. Roitsch, M. C. Gonzalez. "Function and regulation of plant invertases: sweet sensations". Trends Plant Sci. 9, 606-613, 2004. DOI: http://dx.doi.org/10.1016/j.tplants.2004.10.009
  5. A. Sturm. "Invertases: Primary structures, functions, and roles in plant development and sucrose partitioning". Plant Physiol. 121, 1-7. 1999 DOI: http://dx.doi.org/10. 1104/ pp. 121. 1. 1 https://doi.org/10.1104/pp.121.1.1
  6. J. A. Tognetti, H. G. Pontis, G. M. Martinez-Noel. "Sucrose signaling in plants: A world yet to be explored". Plant Signal Behav. 8, e23316. 2013. DOI: http://dx.doi.org/10.4161/psb.23316
  7. H. B. Krishnan, S. G. Pueppke. "Invertases from rust-infected wheat leaves". J Plant Physiol 133, 336-339, 1988. DOI: http://dx.doi.org/10.1016/S0176-1617(88)80211-6
  8. A. Sturm, M. J. Chrispeels. "cDNA cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection". Plant Cell 2, 1107-1119, 1990. DOI: http://dx.doi.org/10.1105/tpc.2.11.1107
  9. L. Zhang, N. Cohn, J. Mitchell. "Induction of a pea cell-wall invertase gene by wounding and its localized expression in phloem". Plant Physiol. 112, 1111-1117, 1996. DOI: http://dx.doi.org/10.1104/ pp.112.3.1111
  10. D. Kim, S. Park, Y. Chung, J. Park, S. Lee, T-K. Lee. "Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings". J. Integr. Plant. Biol. 52, 536-548, 2010. DOI: http://dx.doi.org/10.1111/j.1744-7909.2010.00937.x.
  11. D. Kim, G. Lee, M. Chang, J. Park, Y. Chung, S. Lee, T-K. Lee. "Purification and Biochemical Characterization of Insoluble Acid Invertase (INAC-INV) from Pea Seedlings". J. Agric. Food Chem. 59, 11228-11233, 2011. DOI: http://dx.doi.org/10.1021/jf201057c
  12. N. S. Cohn, J. P. Mitchell. "Immunocytochemical localization of proteins in differentiating tissues of Pisum sativum. Histochemistry 84, 432-438, 1986. DOI: http://dx.doi.org/10.1007/BF00482975
  13. J. W. Stirling, P. S. Graff. "Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium". J Histochem Cytochem 43, 115-123, 1995. DOI: http://dx.doi.org/10.1177/43.2.7529784
  14. M. C. Jamur, C. D. Faraco, L. O. Lunardi, R. P. Siraganian, C. Oliver. "Microwave fixation improves the antigenicity of glutaraldehyde-sensitive antigens while preserving ultrastructural detail". J Histochem Cytochem 43, 307-311, 1995. DOI: http://dx.doi.org/10.1177/43.3.7868860
  15. K. Matsushita, I. Uritani. "Change in invertase activity of sweet potato in response to wounding and purification and properties of its invertases". Plant Physiol 54, 60-66, 1974. https://doi.org/10.1104/pp.54.1.60
  16. N. Benhamou, J. Grenier, M.J. Chrispeels. "Accumulation of beta-fructosidase in the cell walls of tomato roots following infection by a fungal wilt pathogen". Plant Physiol. 97, 739-750. 1991. DOI: http://dx.doi.org/10.1104/pp.97.2.739
  17. T. Roitsch, W. Tanner. "Cell wall invertases: Bridging the gap". Botanica Acta 109, 90-93. 1996. DOI: http://dx.doi.org/10.1111/j.1438-8677.1996.tb00547.x
  18. J. W. Patrick. "Sieve element unloading, cellular pathway, mechanism and control". Physiol Plant 78: 298-308. 1990. DOI: http://dx.doi.org/10.1111/j.1399-3054.1990.tb02095.x
  19. A. S. Tauzin, T. Giardina. "Sucrose and invertases, a part of the plant defense response to the biotic stresses". Frontiers in plant science 293(5), 1-8, 2014. DOI: http://dx.doi.org/10.3389/fpls.2014.00293
  20. Z. Tymowska-Lalanne, M. Kreis. "The plant invertases: physiology, biochemistry and molecular biology". Adv. Bot. Resear. 28, 71-117, 1998. DOI: http://dx.doi.org/10.1016/S0065-2296(08)60294-3