HvIRIP 과발현 유채 형질전환체의 내한성 증진 = Overexpression of Ice Recrystallization Inhibition Protein (HvIRIP) from Barley Enhances Cold Tolerance in Transgenic rapeseed plants

다국어 초록 (Multilingual Abstract)

Rapeseed (Brassica napus) is now the second largest oilseed crop after soybean. Cold temperature tolerance is an important agronomic trait in winter rapeseed that determines the plant's ability to control below freezing temperatures. To improve cold tolerance of rapeseed plants, an expression vector containing an Barley Ice recrystallization inhibition protein (HvIRIP) cDNA driven by a cauliflower mosaic virus 35S promoter was transferred into rapeseed plants. Transgenic expression of HvIRIP was proved by southern- and northern-blot analyses. The level of freezing tolerance of transgenic T3 plants was found to be significantly greater than that of wild-type rapeseed plants by freezing assay.
Proline accumulation during cold stress was also highly induced in the transgenic rapeseed plants. The transgenic plants exhibited considerable tolerance against oxidative damage induced by cold stress. Our results indicated that heterologous HvIRIP expression in transgenic rapeseed plants may induce several oxidative-stress responsive genes to protect from cold stress.

참고문헌 (Reference)

1 노경희, "영산 유채를 이용한 형질전환체 생산" 한국응용생명화학회 54 (54): 26-32, 2011

2 Xin Z, "eskimo1 mutants of Arabidopsis are constitutively freezing-tolerant" 95 : 7799-7804, 1998

3 Brandts JF, "Unfolding and refolding occur much faster for a proline-free protein than for most proline-containing proteins" 74 : 4178-4181, 1977

4 Sakai A, "The role of sugar and related compounds in variations of freezing resistance" 5 : 160-174, 1968

5 Yemm EW, "The estimation of Carbohydrates in plant extracts by anthrone" 57 : 508-514, 1954

6 Gusta LV, "The effect of water, sugars and proteins on the pattern of ice nucleation and propagation in acclimated and nonacclimated canola leaves" 135 : 1642-1653, 2004

7 Rolland F, "Sugar sensing and signaling in plants: conserved and novel mechanisms" 57 : 675-709, 2006

8 Guy CL, "Sucrose phosphate synthase and sucrose accumulation at low temperature" 27 : 507-528, 1992

9 Strauss G, "Stabilization of lipid bilayer vesicles by sucrose during freezing" 83 : 2422-2426, 1986

10 Kaurin, "Seasonal changes in frost hardiness in cloudberry(Rubus chamaemorus)in relation to carbohydrate content with special reference to sucrose" 52 : 310-314, 1981

11 Bates LS, "Rapid determination of free proline for water-stress studies" 39 : 205-207, 1973

12 Booth EJ, "Rapeseeds and rapeseed oil; agronomy, production, and trade. In Rapeseed and Canola oil" Blackwell publishing Ltd. 1-16, 2004

13 Takagi H, "Proline as a stress protectant in yeast: physiological functions, metabolic regulations, and biotechnological applications" 81 : 211-223, 2008

14 Lv WT, "Proline accumulation is inhibitory to Arabidopsis seedlings during heat stress" 156 : 1921-1933, 2011

15 Liu J, "Proline accumulation and salt-stress-induced gene expression in a salt-hypersensitive mutant of Arabidopsis" 114 : 591-596, 1997

16 Thomashow MF, "Plant cold acclimation: freezing tolerance genes and regulatory mechanisms" 50 : 571-599, 1999

17 Fei SZ, "Perennial ryegrass anti-freeze protein genes enhances freezing tolerance in plants" 28 : 129-145, 2008

18 Tremblay K, "Molecular characterization and origin of novel bipartite cold-regulated ice recrystallization inhibition proteins from cereals" 46 : 884-891, 2005

19 Hill LM, "Metabolism of sugars in the endosperm of developing seeds of oilseed rape" 131 : 228-236, 2003

20 Christie P, "Low-temperature accumulation of alcohol dehydrogenase-1 mRNA and protein activity and Maize and Rice seedlings" 95 : 699-706, 1991

21 Jarillo JA, "Low temperature induces the accumulation of alcohol dehydrogenase mRNA in Arabidopsis thaliana, a chilling-tolerant plant" 101 : 833-837, 1993

22 Couee I, "Involvement of soluble sugars in reactive oxygen species balance and responses to oxidative stress in plants" 57 : 449-459, 2006

23 Zamecnik J, "Interaction of antifreeze proteins from coldhardened cereal seedlings with ice nucleation active bacteria" 29 : 718-719, 1992

24 Knight CA, "Inhibition of recrystallization of ice by insect thermal hysteresis proteins: a possible cryoprotective role" 23 : 256-262, 1986

25 Klotke J, "Impact of soluble sugar concentrations on the acquisition of freezing tolerance in accessions of Arabidopsis thaliana with contrasting cold adaptation-evidence for a role of raffinose in cold acclimation" 27 : 1395-1404, 2004

26 John UP, "Ice recrystallization inhibition proteins(IRIPs)and freeze tolerance in the cryophilic Antarctic hair grass Deschampsia antarctica E. Desv" 32 : 336-348, 2009

27 Hightower R, "Expression of antifreeze proteins in transgenic plants" 17 : 1013-1021, 1991

28 Molinari HBC, "Evaluation of the stress-inducible production of proline in transgenic sugarcane (Saccharum spp.): osmotic adjustment, chlorophyll fluorescence and oxidative stress" 130 : 218-229, 2007

29 Moffatt B, "Cold comfort: plant antifreeze proteins" 126 : 5-16, 2006

30 Chew O, "Cold acclimation induces rapid and dynamic changes in freeze tolerance mechanisms in the cryophile Deschampsia antarctica E. Desv" 35 : 829-837, 2012

31 Sasaki H, "Changes in sugar content during cold acclimation and deacclimation of Cabbage seedlings" 78 : 365-369, 1996

32 Zhu GY, "Change in sugar, sterol and fatty acid composition in banana meristems caused by sucrose-induced acclimation and its effects on cryopreservation" 128 : 80-94, 2006

33 Livingston DP, "Carbohydrate partitioning between upper and lower regions of the crown in oat and rye during cold acclimation and freezing" 52 : 200-208, 2006

34 Kankofer M, "Antioxidative defence mechanisms against reactive oxygen species in bovine retained and not-retained placenta: activity of glutathione peroxidase, glutathione transferase, catalase and superoxide dismutase" 22 : 466-472, 2001

35 Atici Ö, "Antifreeze proteins in higher plants" 64 : 1187-1196, 2003

36 Hon WC, "Antifreeze proteins in Winter Rye are similar to pathogenesis-related proteins" 109 : 879-889, 1995

37 DeVries AL, "Antifreeze glycopeptides and peptides: interactions with ice and water" 127 : 293-303, 1986

38 Murashige T, "A revised medium for rapid growth and bioassays with tobacco tissue cultures" 15 : 473-493, 1962

39 Chow PS, "A method for routine measurements of total sugar and starch content in woody plant tissues" 24 : 1129-1136, 2004

40 Meyer K, "A leucine-rich repeat protein of carrot that exhibits antifreeze activity" 447 : 171-178, 1999