Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지

Constitutive Expression of Small Heat Shock Protein Increases Thermotolerance in Transgenic Plant

저 분자량 Heat Shock Protein의 항상적 발현에 의한 형질전판 식물체의 고온내성 증가

  • 이병현 (경북대학교 농과대학 동물자원과학과)
  • Published : 2000.01.01
Download PDF
⟨ Previous Next ⟩

Abstract

To investigate the function of chloroplast small HSP, transgenic tobacco plants (Nicotiana tabacum L. cv. Samsun) that constitutively overexpress the chloroplast small HSP (NtHSP21) from N. tabacum cv. Petit Havana SR1 were generated. Five homozygous lines of transformants showing different constitutive expression levels of the NtHSP21 were selected. To determine whether constitutive overexpression of NtHSP21 protein affects thermotolerance, wild-type and transformants were grown in Petri dishes, heat-stressed at 52$^{\circ}C$ for 45 min, and then incubated in normal growth condition. When heat-stressed wild-type plantlets were incubated at $25^{\circ}C$, leaf color gradually became white and all trio plantlets finally died within a week. As for the transformants, however, more than 70% of them remained green and survived under the conditions in which all the wild-type plants were dying. It was also found that the levels of NtHSP21 were correlated with the degree of thermotolerance. These results suggest that the NtHSP21 protein in transformants is responsible for the increase in thermotolerance.

고등식물에 있어서 엽록체에 존재하는 저 분자량 HSP의 기능을 밝히기 위하여 담배 (Nicotina tabacum cv. Petit Havana SRI)로부터 분리한 cDNA (NtHSP21)를 도입한 형질전환 담배 식물체를 재생하였다. 상온에서의 발현량이 서로 다른 5개의 순계 형질전환 식물체를 선발하였다 상온에서 발현된 엽록체 small HSP가 식물의 고온내성에 미치는 영향을 조사하기 위하여 기내에서 생장시킨 유식물을 52$^{\circ}C$에서 45분간 열처리한 후 생장온도에서의 변화를 조사하였다. 그 결과 wild-type식물의 경우 1주일 이내에 모두 고사하였으나 형질전환 식물체의 약 70%는 생존하였다. 또한 이러한 고온내 성은 상온에서 발현된 단백질의 양에 비례하여 증감하였다. 따라서 엽록체에 존재하는 small HSP가 식물의 고온내성 획득에 있어서 중요한 역할을 담당할 것으로 사료된다.

Keywords

References

  1. Plant J v.13 Accumulation of small heat shock proteins, including mitochondrial HSP22, induced by oxidative stress and adaptive response in tomato cells Banzet N;Richaud C;Deveaux Y;Kazumaier M;Gagnon J;Triantaphylides C
  2. Mol Gen Genet v.226 Analysis of conserved domains identifies a unique structural feature of a chloroplast heat shock protein Chen Q;Vierling E
  3. Plant Physiol v.89 Thermotolerance of isolated mitochondria associated with heat shock proteins Chou M;Chen Y;Lin C
  4. FEBS Lett v.430 The mitochondrial small heat shock protein protects NADH:ubiquinone oxidoreductase of the electron transport chain during heat streaa in plants Downs CA;Heckathorn SA
  5. Eur J Biochem v.173 Temperature-dependent binding to the thylakoid membranes of nuclear-coded chloroplast heat shock proteins Glaczinski H;Kloppsteck K
  6. Plant Physiol v.116 The small, methionine-rich chloroplast heat-shock protein protects photosystem Ⅱelectron transport during heat stress Heckathorn SA;Downs CA;Sharkey TD;Cleman JS
  7. Science v.227 A simple and general method for transferring genes into plants Horsch RB;Fry JE;Hofman NL;Eichholz D;Rogers SG;Fraley RT
  8. Plant Mol Biol v.37 Evolutionary origin of two genes for chloroplast small heat shock protein of tobacco Lee BH;Tanaka Y;Iwasaki T;Yamamoto N;Kayano T;Miyao M
  9. J Biol Chem v.270 Structure and in vitro molecular chaperone activity of cytosolic small heat shock proteins from pea Lee GJ;Pokala N;Vierling E
  10. Ann Rev Genet v.22 The heat shock proteins Lindquist S;Craig EA
  11. Methods in Molecular Biology v.2 RNA extraction by the guanidine thiocyanate procedure McGookin R;Walker JM(ed.)
  12. Physiol Plant v.15 A revise medium for rapid growth and bioassays with tobacco tissue cultures Murashige T;Skoog F
  13. Nucl Acids Res v.8 Rapid isolation of high molecular weight plant DNA Murray MG;Tompson WF
  14. Plant Physiol v.93 Cloning, sequence analysis, expression of a cDNA encoding a plastid-localized heat shock protein in maize Nieto-Sotelo J;Vierling E;David Ho T-H
  15. NATO ASI series, Biochemical and Cellular Mechanisms of Stress Tolerance in Plants v.H86 Studies of a chloroplast-localized small heat shock protein in Arabidopsis Osteryoung KW;Pipes B;Wehmeyer N;Vierling E;Cherry JH(ed.)
  16. Plant Physiol v.111 Heat-shock response in heat-tolerant and nontolerant variants of Agrosits palustris Huds Park S;Shivaji R;Krans JV;Luthe DS
  17. Ann Rev Genet v.27 The function of heat-shock proteins in stress tolerance: degradation and reactivation of proteins Parsell DA;Lindquist S
  18. Molecular Cloning: A Laboratory Manual, 2nd ed. Sambrook J;Fritsch EF;Maniatis T
  19. EMBO J v.7 Evidence for protection by heat-shock proteins against photoinhibition during heat shock Schuster G;Even D;Kloppstech K;Ohad I
  20. Annu Rev Plant Physiol Plant Mol Biol v.42 The roles of heat shock proteins in plants Vierling E
  21. EMBO J v.7 A heat shock protein localized to chloroplasts is a member of an eukaryotic superfamily of heat shock proteins Vierling E;Nagao RT;DeRocher AE;Harris LM
  22. Genetics v.141 The molecular evolution of the small heat-shock proteins in plants Waters ER
  23. J Exp Bot v.47 Evolution, structure and function of the small heat shock proteins in plants Waters ER;Lee GJ;Vierling E