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1 Chinese Journal of Eco-Agriculture, July 2009, 17(4): DOI: /SP.J 郑国华 1 张贺英 1 钟秀容 2 ( ) 以盆栽解放钟嫁接的枇杷结果小苗 (Eriobotrya japonica Lindl.) 为试材, 采用人工降温的方法, 研究低温胁迫下枇杷叶片细胞超微结构及膜透性和保护酶活性的变化结果表明, 7 2 低温处理时, 枇杷叶片大多数细胞器基本无损, 排列有序, PMP MDA 含量先上升后下降, 保护酶活性先增大后减小, 表明枇杷小苗对 7 2 低温逆境有一定的适应能力 ; 2 低温胁迫叶绿体双层膜结构破坏严重, 类囊体垛叠程度很低, 形不成典型的基粒, 还可见线粒体双层膜结构, 但己经没有明显的内脊, 内部呈小泡化, PMP MDA 含量呈一直上升的趋势, 保护酶活性受到抑制, 表明枇杷结果小苗已经受到冻害 ; 7 低温胁迫细胞质膜已经破裂, 原生质体浓缩, 叶绿体扭曲变形相互融合, 液泡破裂, 线粒体膜结构受损, 脊消失, PMP MDA 含量大幅度增加, 保护酶活性受到严重抑制, 表明枇杷营养生长已经受到极为严重的伤害由此可见, 枇杷结果小苗在 0 以下受冻严重, 营养生长受阻, 从而影响枇杷正常生长以及产量品质的提高低温胁迫枇杷叶片细胞超微结构膜透性保护酶活性 : S667.3 : A : (2009) Changes in cell ultra-structure, membrane permeability and protective enzyme activity in Eriobotrya japonica Lindl. leaves under cold stress ZHENG Guo-Hua 1, ZHANG He-Ying 1, ZHONG Xiu-Rong 2 (1. College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou , China; 2. Pathology Department, Fujian Medical University, Fuzhou , China) Abstract Using a pot cultured Eriobotrya japonica Lindl. grafted in Jiefangzhong as an experimental material, changes in E. japonica leaf cell ultra-structure, membrane permeability and protective enzyme activity were determined. The results show that under controlled temperature of 7 and 2, most leaf cell organelles of E. japonica are not injured and orderly arrange. PMP and MDA content initially raises and then falls, so also is the protective enzyme activity. This suggests that E. japonica can adapt to chilling temperatures of 7 and 2. Under 2, leaf cell chloroplasts are destroyed. Thylakoid is not packed to its normal grana and therefore fails to form typical structures. Though the mitochondrion membrane structure can still be visible, its cristae number drops. Because of increasing content of PMP and MDA, protective enzyme activity is inhibited. Hence the leaf of E. japonica suffers severe frostbite. Under a temperature stress of 7, plasma membranes break up and protoplasts become greatly concentrated. In fact chloroplasts entirely decompose. Grana lamellace twists, disfigures and even fuses. Mitochondrion membrane system is completely destroyed and cristae can not be found. PMP and MDA content raises drastically, hence protective enzyme activity is greatly inhibited. The vegetation growth of E. japonica are badly distorted. In summary, E. japonica leaf severely freezes when temperatures fall below zero, which inhibits the vegetation growth of the plant. Key words Cold stress, Eriobotrya japonica Lindl., Leaf, Cell ultra-structure, Membrane permeability, Protective enzyme activity (Received July 20, 2008; accepted Nov. 30, 2008) (1965~),,,, fafuzgh@126.com : :

2 (Eriobotrya japonica Lindl.),,,,, [1], [2 5], [6,7],,, (), 1 材料与方法 ,,,, 5,, 20~10,, 0.5,,, 10 h, Lx (15 )5, 3 12 h 24 h 48 h 72 h, 3,, 72 h 7 6 h 12 h 18 h 24 h,, 3,, 24 h 1.2 5, 2 mm 2 mm, ph 7.2 5%,, (spurr ) h, 48 h,, LKB8800 ( 10 µm), JEM1010 (SOD)(POD) [8], (CAT) Aebi [9], (APX) Nakano Asada [10], (MDA) [11], (PMP) [12] SAS 结果与分析 , 15,, 1 3~5,,,,,,,,,,,,,,,, [ 1(1) (3)] , 7,,,,,,, [ 1(4) (7)] ,,,,,,,,, [ 1(8) (11)] ,,,,

3 4 : 741 Fig. 1 图 1 低温胁迫下枇杷叶片细胞超微结构的变化 Change of cell ultrastructure of E. japonica leaves under cold stress (1) ( 20K); (2) ( 40K); (3) ( 60K); (4)7 ( 15K); (5)7 ( 50K); (6)7 ( 80K); (7)7 ( 6K); (8)2 ( 20K); (9)2 ( 20K); (10)2 ( 80K); (11)2 ( 8K); (12) 2 ( 15K); (13) 2 ( 50K); (14) 2 ( 80K); (15) 2 ( 3K); (16) 7 ( 15K); (17) 7 ( 80K); (18) 7 ( 3K); Ch: ; Cl: ; CW: ; M: ; Pm: ; Sg: ; T: ; V: ; Lg: (1)Control chloroplast( 20K); (2)Control chloroplast grana lamellace( 40K); (3)Control mitochondria( 60K); (4)7 chloroplast( 15K); (5)7 chloroplast grana lamellace( 50K); (6)7 mitochondria( 80K); (7)7 cell( 6K); (8)2 plasmolemma, chloroplast grana lamellace( 20K); (9)2 chloroplast( 20K); (10)2 mitochondria( 80K); (11)2 cell( 8K); (12) 2 chloroplast, starch grain( 15K); (13) 2 chloroplast grana lamellace( 50K); (14) 2 mitochondria( 80K); (15) 2 cell( 3K); (16) 7 chloroplast( 15K); (17) 7 mitochondria( 80K); (18) 7 cell( 3K); Ch: Chloroplast; Cl: Chloroplast lamellae; Cw: Cell wall; M: Mitochondria; Pm: Plasmolemma; Sg: Starch grain; T: Tonoplast; V: Vacuole; Lg: Liposome grain.

742 2009 17,,,,,, ;,, ; [ 1(12) (15)] 2.1.5 7,, ;,,, ;, ;, ;,,,,,,, ;, [ 1(16) (18)] 2.2 PMP 7 2 PMP, 7 24 h PMP, 24 h PMP, 17.38%, 2 12 h PMP, 46.48%; 2 PMP, 72 h PMP, 57.76% SAS6.

4 ,,,,,, ;,, ; [ 1(12) (15)] ,, ;,,, ;, ;, ;,,,,,,, ;, [ 1(16) (18)] 2.2 PMP 7 2 PMP, 7 24 h PMP, 24 h PMP, 17.38%, 2 12 h PMP, 46.48%; 2 PMP, 72 h PMP, 57.76% SAS6.04, CK 7 2 PMP, 2 ( 1) 2, 7 PMP, PMP 6 h, %,, 24 h, PMP % SAS6.04, PMP, MDA MDA 2, 7 2 MDA 图 2 7 低温胁迫对枇杷叶片质膜相对透性的影响 Fig. 2 Effect of 7 cold stress on membrane permeability in E. japonica, 7 24 h MDA, 26.19%, 2 12 h MDA, 55.95%, SAS6.04, 72 h MDA 2 MDA, 12 h, 12 h, 72 h MDA %, SAS SOD 2, 7 2 SOD, 24 h 12 h, 10.46% 11.78% 2 12 h SOD, 13.13%, SOD, 48 h, 72 h SOD 11.26% SAS6.04, 2, h, 2 SOD, POD 2 POD 7 2 2,, 7 POD 24 h, 19.82%, 2 12 h 表 1 低温胁迫对枇杷叶片质膜相对透性 (PMP) 的影响 Tab. 1 Effect of cold stress on plasma membrane permeability in leaves of E. japonica (PMP) Plasma membrane permeability (%) Treatment 12 h 24 h 48 h 72 h CK 29.69±1.16Aa 29.69±1.16Aa 29.69±1.16Aa 29.69±1.16Aa ±1.35Bb 34.85±1.11Aab 32.73±1.41Aa 30.02±1.52Aa ±1.28Cc 38.97±1.87Aab 35.15±0.94Aa 30.77±1.15Aa ±1.82Dd 45.00±1.52Bc 45.74±1.04Bb 46.84±1.67Bb

5 4 : 743 Treatment Tab. 2 Item 表 2 低温胁迫对枇杷叶片 MDA 含量及保护酶活性的影响 Effect of cold stress on MDA content and protective enzyme activity in leaves of E. japonica MDA MDA content [mg (g 1 )FW] and protective enzyme activity [unit mg 1 (pro) min 1 ] 12 h 24 h 48 h 72 h CK MDA 0.84±0.04Aa 0.84±0.04Bc 0.84±0.04Ab 0.84±0.04Aa SOD ±2.11Aa ±2.11Aa ±2.11Cc ±2.11Aa POD 48.14±2.33Aa 48.14±2.33Aa 48.14±2.33Aa 48.14±2.33Aa CAT 84.82±0.95Aa 84.82±0.95Aa 84.82±0.95Aa 84.82±0.95Aa APX ±3.51Aa ±3.51Aa ±3.51Aa ±3.51Aa 7 MDA 0.66±0.07Aa 1.06±0.09Bb 1.04±0.14Aab 1.00±0.06Aa SOD ±1.05Bb ±1.25Bb ±4.43Bb ±2.39Aa POD 54.94±1.75Bb 57.68±2.14Bb 53.01±2.09Bb 49.47±2.01Aa CAT 93.74±1.15Bb ±1.28Bb 93.42±1.11Aa 90.97±1.03Aa APX ±4.05Bb ±5.25Bb ±4.13Bb ±3.69Aa 2 MDA 1.31±0.14Bb 1.08±0.07Bb 1.05±0.04Aab 1.02±0.05Aa SOD ±2.20Bb ±1.47Bb ±2.51Aa ±3.47Aa POD 58.23±2.41Cc 57.89±2.27Bb 54.29±2.19Bb 50.19±2.13Aa CAT ±2.01Cc ±1.57Cc ±1.24Bb ±1.17Aa APX ±5.19Cc ±4.79Bb ±4.51Bb ±4.27Aa 2 MDA 1.33±0.23Bb 1.35±0.06Aa 1.45±0.40Aa 1.69±0.12Bb SOD ±1.26Cc ±3.71Cc ±3.42Cc ±2.31Bb POD 74.35±2.76Dd 65.41±2.59Cc 57.72±2.38Cc 46.38±2.22Ab CAT ±2.26Dd ±1.71Bb 79.41±1.32Aa 56.73±0.91Bb APX ±6.08Dd ±3.41Aa ±3.14Cc ±2.89Bb, 20.96%, 2 12 h POD, 54.45%; 2 POD, POD, 72 h POD, 3.66% SAS6.04, 2, h CAT 2, CAT 7 2 2,, 2 7 CAT 12 h 24 h, 47.30% 21.02%, 72 h, CAT, ; 2 12 h CAT, 96.85%, CAT, 48 h CAT, 72 h 33.12%; SAS6.04, 2, h APX 2, APX 7 2 2,, 7 24 h APX, 29.47%, 2 12 h, 37.13% 72 h 7 2 APX 2 12 h APX, 90.14%, APX, 48 h APX, 72 h APX 24.76% SAS6.04, 2 72 h, h MDA 3, 7,,, MDA, 24 h, %, SAS6.04, 7 SOD CAT POD APX, 6 h, 13.80% % 68.59% 92.34%,, 24 h 25.71% 41.85% 17.93% 51.68%, SAS6.04

6 表 3 7 低温胁迫对枇杷叶片 MDA 含量及保护酶活性的影响 Tab. 3 Effect of 7 cold stress on MDA content and protective enzyme activity in leaves of E. japonica Lindl. Treatment MDA SOD CAT POD APX [mg g 1 (FW)] [unit mg 1 (pro) min 1 ] CK 0.84±0.04Ee ±2.11Bb 84.82±0.95Aa 48.14±2.33BCd ±3.51 Aa 6 h 1.19±0.05Dd ±1.02Aa ±4.38Bb 81.16±1.63Aa ±8.32Bb 12 h 1.54±0.07Cc ±2.14Bb ±2.07 Cc 71.00±3.36ABb ±5.70Cc 18 h 1.89±0.06Bb ±2.39Cc 98.35±3.40 Aa 60.42±1.64Bc ±4.39Aa 24 h 2.30±0.09Aa ±2.43Dd 49.32±2.51Dd 39.51±4.67Cd 64.57±3.79Dd 7, 3 小结与讨论 [13,14],,, ,, 7 2,,,, 7 2, 2,,,,,,,, 2 7,,,,,,,,,, 7,,,, [15] [16] O 2 [17], [13], [18,19] ;,,,,,,, ;,,,,,,,, ( ),,, h MDA PMP, 2 7, 7 MDA PMP,,,,,,, [17], 7 2 SOD POD CAT APX,,, 72 h,,,, ; 2,,,,,, ; 7 6 h, 6 h,,

7 4 : h,,,,,,,,,,, 参考文献 [1],. [M]. :, 1996: [2],,,. Ca 2+ [J]., 2000, 18(2): [3],,. [J]., 2003, 15(4): [4],,,. [J]., 2004, 33(1): [5],. [J]., 2005, 4(1): [6],. [J]., 2008, 15(2): [7],,,. [J]., 2008, 34(2): [8],,. [M]. :, 1990: 37 39, [9] Aebi H. Catalase in vitro[j]. Methods in Enzymology, 1984, 105: [10] Nakano Y., Asada K. Hydrogen eproxide is scavenged by ascobate-specific peroxidase in spinach chloroplasts[j]. Plant Cell Physiol, 1981, 22: [11]. [M]. :, 1999: [12],,. [M]. :, 2000: [13]. [J]., 1992, 9(3): [14],,,. [J]., 2000, 42(8): [15],. [J]., 2001, 30(2): [16],,,. [J]., 2003, 39( 1): [17] Navari-Izzo F., Rascid N. Plant responses to water-deficit conditions[m] Pessarakli M. Handbook of Plant and Crop Stress (2 nd edition). New York: Marcel Dekker, 1999: [18] Nilsen E. T., Orcutt D. M. The Physiology of Plants under Stress[M]. New York: John Wiley & Sons, 1996: 70 [19] Alscher R. G., Erturk N., Heath L. S. Role of superoxide dismutases (SOD) in controlling oxidatives stress in plants[j]. Journal of Experimental Botany, 2002, 53:

Untitiled

Untitiled 中国农学通报 2009,25(11):117-122 Chinese gricultural Science Bulletin 钙处理对枇杷果实采后生理特性的研究郑国华 ( 福建农林大学园艺学院, 福州 350002) 摘要 : 适当的钙处理可明显减缓枇杷果实的采后生理活力试验结果