西瓜感染黄瓜绿斑驳花叶病毒后糖分代谢与倒瓤的关系
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摘要
为揭示人工接种黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)后西瓜糖分代谢与倒瓤的关系,采用高效液相色谱法测定了接种CGMMV后西瓜的葡萄糖、果糖、蔗糖含量变化,采用比色法测定了糖酵解途径关键酶及蔗糖代谢相关酶活性的变化。结果显示,西瓜苗感染CGMMV后,果实中葡萄糖、果糖、蔗糖含量均较对照发生明显变化,接种处理后葡萄糖含量在授粉7~28 d显著高于对照,之后迅速下降,35 d时显著低于对照,仅为对照的24.87%;果糖含量仅在授粉后7~14 d高于对照,21~35 d显著低于对照;接种处理和对照的蔗糖含量在授粉后14~35 d均逐渐上升,但14~35 d接种处理显著低于对照。接种处理后己糖激酶、磷酸果糖激酶和果糖激酶活性均在授粉后7~14 d显著低于对照,21~28 d显著高于对照,35 d时显著低于对照;丙酮酸激酶活性授粉后7~21 d显著低于对照,35 d时显著高于对照;酸性转化酶和中性转化酶活性在授粉后7~21 d显著低于对照,35 d时高于对照;蔗糖磷酸合成酶和蔗糖合成酶活性均显著低于对照,其中蔗糖合成酶活性在授粉后28 d时仅为对照的21.32%。推断西瓜苗接种CGMMV后糖酵解的4个关键酶协同作用,调控糖酵解,进而调控葡萄糖和果糖的积累与代谢,同时蔗糖代谢相关酶活性降低,阻碍蔗糖合成,即西瓜苗感染CGMMV导致倒瓤与其糖分的合成与代谢密切相关。
To reveal the relationships between the variation of sugar contents, the activities of the key enzymes in glycometabolism and blood-flesh of watermelon after inoculated by Cucumber green mottle mosaic virus(CGMMV), the contents of glucose, fructose and sucrose were analyzed with high-performance liquid chromatography and the activities of the key glycolytic enzymes and sucrose-metabolizing enzymes were measured with colorimetry. The results showed that the content of glucose, fructose and sucrose changed obviously compared with the control group. After inoculated by CGMMV, the content of glucose was significantly increased 7-28 d after pollination compared to that of the control, then reduced rapidly and was only 24.87% of the control at the 35 th day after pollination. The content of fructose was higher than the control 7-14 d after pollination, then decreased obviously, and it was significantly lower than the control after 21-35 d. In all cases, the content of sucrose increased in 14 to 35 d with the growth of watermelon. However, compared with the control, the sucrose content of watermelon inoculated with CGMMV was significantly lower after 14-35 d. After inoculation, the activities of hexokinase, phosphofructokinase and fructokinase were significantly lower than that of the control after7-14 d and at the 35 th day after pollination, and were significantly higher than the control after 21-28 d.The activity of pyruvate kinase was significantly lower than that of control 7-21 d after pollination, and significantly higher than the control at the 35 th day. The results of acid invertase and neutral invertase assay showed that the activities of these two enzymes were significantly lower than that of the control7-28 d after pollination, and it was higher than the control at the 35 th day. The activities of sucrose synthase and sucrose phosphate synthase were significantly lower than that of control during the test, and the activity of sucrose synthase was only 21.32% of the control at the 28 th day after pollination. It was concluded that the four key enzymes of EMP had synergistic effect with each other, and played an important role in the regulation of glycolysis and the accumulation of glucose and fructose after inoculated by CGMMV. The acid invertase, neutral invertase, sucrose synthase and sucrose phosphate synthase played an important role in the synthesis of sucrose, and the change of them may prevent the synthesis of sucrose. In summary, synthesis and metabolism of sugar was closely related to the blood-flesh of watermelon caused by CGMMV.
To reveal the relationships between the variation of sugar contents, the activities of the key enzymes in glycometabolism and blood-flesh of watermelon after inoculated by Cucumber green mottle mosaic virus(CGMMV), the contents of glucose, fructose and sucrose were analyzed with high-performance liquid chromatography and the activities of the key glycolytic enzymes and sucrose-metabolizing enzymes were measured with colorimetry. The results showed that the content of glucose, fructose and sucrose changed obviously compared with the control group. After inoculated by CGMMV, the content of glucose was significantly increased 7-28 d after pollination compared to that of the control, then reduced rapidly and was only 24.87% of the control at the 35 th day after pollination. The content of fructose was higher than the control 7-14 d after pollination, then decreased obviously, and it was significantly lower than the control after 21-35 d. In all cases, the content of sucrose increased in 14 to 35 d with the growth of watermelon. However, compared with the control, the sucrose content of watermelon inoculated with CGMMV was significantly lower after 14-35 d. After inoculation, the activities of hexokinase, phosphofructokinase and fructokinase were significantly lower than that of the control after7-14 d and at the 35 th day after pollination, and were significantly higher than the control after 21-28 d.The activity of pyruvate kinase was significantly lower than that of control 7-21 d after pollination, and significantly higher than the control at the 35 th day. The results of acid invertase and neutral invertase assay showed that the activities of these two enzymes were significantly lower than that of the control7-28 d after pollination, and it was higher than the control at the 35 th day. The activities of sucrose synthase and sucrose phosphate synthase were significantly lower than that of control during the test, and the activity of sucrose synthase was only 21.32% of the control at the 28 th day after pollination. It was concluded that the four key enzymes of EMP had synergistic effect with each other, and played an important role in the regulation of glycolysis and the accumulation of glucose and fructose after inoculated by CGMMV. The acid invertase, neutral invertase, sucrose synthase and sucrose phosphate synthase played an important role in the synthesis of sucrose, and the change of them may prevent the synthesis of sucrose. In summary, synthesis and metabolism of sugar was closely related to the blood-flesh of watermelon caused by CGMMV.
引文
Adzhemyan LA,Gevorkyan ZG,Amirkhanyan VA.1976.Changes in sugars and free amino acids in cucumber during infection by some virus diseases.Biologicheskii Zhurnal Armenii,29(1):91-94
Al-Shahwan IM,Abdalla OA.1992.A strain of Cucumber green mottle mosaic virus(CGMMV)from bottlegourd in Saudi Arabia.Journal of Phytopathology,134(2):152-156
Ali A,Natsuaki T,Okuds S.2004.Identification and molecular characterization of viruses infecting cucurbits in Pakistan.Journal of Phytopathology,152(11):677-682
Brown AC Jr.,Summer WL.1985.Carbohydrate accumulation and color development in watermelon.Journal of the American Society for Horticultural Science,110(5):683-687
Célix A,Luis-Arteaga M,Rodríguez-Cerezo E.1996.First report of Cucumber green mottle mosaic tobamo virus infecting greenhouse-grown cucumber in Spain.Plant Disease,80(11):1303
Chen HY,Lin SM,Chen Q,Zhao WJ,Liao FR,Chen HJ,Zhu SF.2009.Complete genomic sequence of a watermelon isolate of Cucumber green mottle mosaic virus in northern China.Chinese Journal of Virology,25(1):68-72(in Chinese)[陈红运,林石明,陈青,赵文军,廖富荣,陈洪俊,朱水芳.2009.黄瓜绿斑驳花叶病毒辽宁分离物全基因组序列测定.病毒学报,25(1):68-72]
Chu SP,Gong WR,Hu J,Ding ZK,Cheng ZB,Tian ZH.2014.The situation of occurrence and control on the CGMMV in Jiangsu area.Jiangsu Agricultural Sciences,42(12):183-184(in Chinese)[褚姝频,龚伟荣,胡婕,丁志宽,程兆榜,田子华.2014.江苏地区黄瓜绿斑驳花叶病毒病的发生概况与防控技术.江苏农业科学,42(12):183-184]
Gao JF.2000.Plant physiology experiment technology.Xi’an:World Publishing Corporation,pp.146(in Chinese)[高俊凤.2000.植物生理学实验技术.西安:世界图书出版公司,pp.146]
Gao ZF,Schaffer AA.1999.A novel alkalineα-galactosidase from melon fruit with a substrate preference for raffinose.Plant Physiology,119(3):979-988
Guo ZF,Li MQ.1993.Partial purification and regulatory properties of the cytoplasmic pyruvate kinase from Passiflora edulis leaves.Acta Phytophysiologica Sinica,19(2):105-110(in Chinese)[郭振飞,李明启.1993.鸡蛋果叶片细胞质丙酮酸激酶的部分纯化及其调节特性.植物生理学报,19(2):105-110]
Huber SC,Akazawa T.1986.A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells.Plant Physiology,81(4):1008-1013
Jang JC,Sheen J.1994.Sugar sensing in higher plants.Plant Cell,6(11):1665-1679
Kim SM,Lee JM,Yim KO,Oh MH,Park JW,Kim KH.2003.Nucleotide sequences of two Korea isolates of Cucumber green mottle mosaic virus.Molecules and Cells,16(3):407-412
Li LM,Wu YH,Zhao XX,Wang WH,Wang L,Cai M.2010.Studies on the yield,quality and virulence ratio in seeds of watermelon inoculated by Cucumber green mottle mosaic virus.Plant Protection,36(6):82-86(in Chinese)[李立梅,吴元华,赵秀香,王文航,王林,蔡明.2010.黄瓜绿斑驳花叶病毒对西瓜产量、品质及种子带毒的影响.植物保护,36(6):82-86]
Li W,Shao YZ,Chen WX.2005.Research advances in the relationships of sucrose phosphate synthase(SPS)with fruit quality and maturity and senescence.Acta Botanica Boreali-Occidentalia Sinica,25(11):2340-2344(in Chinese)[李雯,邵远志,陈维信.2005.蔗糖磷酸合成酶(SPS)与果实品质及成熟衰老的研究进展.西北植物学报,25(11):2340-2344]
Lin Y,Yang YB,Wang C,Wang WH,Mao LR.2012.A preliminary report on Cucumber green mottle mosaic virus in watermelon of Wentai area.Journal of Zhejiang Agricultural Sciences,(1):83-85(in Chinese)[林燚,杨瑜斌,王驰,王文华,毛玲荣.2012.温台地区西瓜发生黄瓜绿斑驳花叶病毒调查初报.浙江农业科学,(1):83-85]
Liu HY,Zhu ZJ,Qian QQ,Ge ZP.2004.The effects of different rootstocks on the sugar metabolism and related enzyme activities in small and early-maturing watermelon during fruit development.Acta Horticulturae Sinica,31(l):47-52(in Chinese)[刘慧英,朱祝军,钱琼秋,葛志平.2004.砧木对小型早熟西瓜果实糖代谢及相关酶活性的影响.园艺学报,31(l):47-52]
Liu Y,Peng B,Yang XH,Li XD,Gu QS.2012.Detection of Cucumber green mottle mosaic virus of watermelon blood flesh in Jinan area.China Vegetables,(10):75-79(in Chinese)[刘颖,彭斌,杨晓红,李向东,古勤生.2012.济南地区倒瓤西瓜CGMMV的检测.中国蔬菜,(10):75-79]
Park GS.2001.Occurrence of two tobamovirus diseases in cucurbits and control measures in Korea.The Plant pathology Journal,17(5):243-248
Qian QQ,Liu HY,Zhu ZJ.2004.Studies on sugar metabolism and related enzymes activity during watermelon fruit development as influenced by grafting.Journal of Zhejiang University(Agriculture and Life Sciences),30(3):285-289(in Chinese)[钱琼秋,刘慧英,朱祝军.2004.嫁接西瓜果实发育过程中蔗糖代谢及相关酶调控的研究.浙江大学学报(农业与生命科学版),30(3):285-289]
Ruan YL,Chourey PS,Delmer DP,Perez-Grau L.1997.The differential expression of sucrose synthase in relation to diverse patterns of carbon partitioning in developing cotton seeds.Plant Physiology,115(2):375-385
Schaffer AA,Petreikov M.1997.Inhibition of fructokinase and sucrose synthase by cytosolic levels of fructose in young tomato fruit undergoing transient starch synthesis.Physiologia Plantarum,101(4):800-806
Srivastava AK,Tiwari CB.1998.Polyphenoloxidase activity as influenced by CGMMV infected cucumber.Journal of Living World,5(1):7-10
Stadler R,Truermit E,Gahrtz M,Sauer N.1999.The At SUC1 sucrose carrier may represent the osmotic driving force for anther dehiscence and pollen tube growth in Arabidopsis.The Plant Journal,19(3):269-278
Tian YL,Liu DM,Zhang YJ,Li MF,Ma ZH.2009.Bioassay and genomic studies on the two isolates of Cucumber green mottle mosaic virus from Beijing and Shandong.Plant Quarantine,23(6):1-6(in Chinese)[田永蕾,刘冬梅,张永江,李明福,马占鸿.2009.黄瓜绿斑驳花叶病毒北京和山东分离物的生物学测定及其基因组比较.植物检疫,23(6):1-6]
Ugaki M,Tomiyama M,Kakutani T,Hidaka S,Kiguchi T,Nagata R,Sato T,Motoyoshi F,Nishiguchi M.1991.The complete nucleotide sequence of Cucumber green mottle mosaic virus(SH strain)genomic RNA.Journal of General Virology,72(7):1487-1495
Vani S,Varma A.1993.Properties of Cucumber green mottle mosaic virus isolated from water of river Jamuna.Indian Phytopathology,46(2):118-122
Wang YZ,Zhang DP.2000.Regulating effects of ethylene on carbohydrate metabolism in‘Starkrimson’apple fruit during the ripening period.Acta Horticulturae Sinica,27(6):391-395(in Chinese)[王永章,张大鹏.2000.乙烯对成熟期新红星苹果果实碳水化合物代谢的调控.园艺学报,27(6):391-395]
Xue YL.1985.Laboratory manual of plant physiology.Shanghai:Shanghai Science and Technology Press(in Chinese)[薛应龙.1985.植物生理学实验手册.上海:上海科学技术出版社]
Yakushiji H,Nonami H,Fukuyama T,Ono S,Takagi N,Hashimoto Y.1996.Sugar accumulation enhanced by osmoregulation in Satsuma mandarin fruit.Journal of the American Society for Horticultural Science,121:466-472
Zhou LL,Wu YH,Zhao XX,Li LM,Cai M,Wang L,Wang WH.2008.The biological characteristics of Cucumber green mottle mosaic virus and its effects on yield and quality of watermelon.Journal of Shenyang Agricultural University,39(4):417-422(in Chinese)[周玲玲,吴元华,赵秀香李立梅,蔡明,王林,王文航.2008.黄瓜绿斑驳花叶病毒生物学特性及对西瓜生长的影响.沈阳农业大学学报,39(4):417-422]
Al-Shahwan IM,Abdalla OA.1992.A strain of Cucumber green mottle mosaic virus(CGMMV)from bottlegourd in Saudi Arabia.Journal of Phytopathology,134(2):152-156
Ali A,Natsuaki T,Okuds S.2004.Identification and molecular characterization of viruses infecting cucurbits in Pakistan.Journal of Phytopathology,152(11):677-682
Brown AC Jr.,Summer WL.1985.Carbohydrate accumulation and color development in watermelon.Journal of the American Society for Horticultural Science,110(5):683-687
Célix A,Luis-Arteaga M,Rodríguez-Cerezo E.1996.First report of Cucumber green mottle mosaic tobamo virus infecting greenhouse-grown cucumber in Spain.Plant Disease,80(11):1303
Chen HY,Lin SM,Chen Q,Zhao WJ,Liao FR,Chen HJ,Zhu SF.2009.Complete genomic sequence of a watermelon isolate of Cucumber green mottle mosaic virus in northern China.Chinese Journal of Virology,25(1):68-72(in Chinese)[陈红运,林石明,陈青,赵文军,廖富荣,陈洪俊,朱水芳.2009.黄瓜绿斑驳花叶病毒辽宁分离物全基因组序列测定.病毒学报,25(1):68-72]
Chu SP,Gong WR,Hu J,Ding ZK,Cheng ZB,Tian ZH.2014.The situation of occurrence and control on the CGMMV in Jiangsu area.Jiangsu Agricultural Sciences,42(12):183-184(in Chinese)[褚姝频,龚伟荣,胡婕,丁志宽,程兆榜,田子华.2014.江苏地区黄瓜绿斑驳花叶病毒病的发生概况与防控技术.江苏农业科学,42(12):183-184]
Gao JF.2000.Plant physiology experiment technology.Xi’an:World Publishing Corporation,pp.146(in Chinese)[高俊凤.2000.植物生理学实验技术.西安:世界图书出版公司,pp.146]
Gao ZF,Schaffer AA.1999.A novel alkalineα-galactosidase from melon fruit with a substrate preference for raffinose.Plant Physiology,119(3):979-988
Guo ZF,Li MQ.1993.Partial purification and regulatory properties of the cytoplasmic pyruvate kinase from Passiflora edulis leaves.Acta Phytophysiologica Sinica,19(2):105-110(in Chinese)[郭振飞,李明启.1993.鸡蛋果叶片细胞质丙酮酸激酶的部分纯化及其调节特性.植物生理学报,19(2):105-110]
Huber SC,Akazawa T.1986.A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells.Plant Physiology,81(4):1008-1013
Jang JC,Sheen J.1994.Sugar sensing in higher plants.Plant Cell,6(11):1665-1679
Kim SM,Lee JM,Yim KO,Oh MH,Park JW,Kim KH.2003.Nucleotide sequences of two Korea isolates of Cucumber green mottle mosaic virus.Molecules and Cells,16(3):407-412
Li LM,Wu YH,Zhao XX,Wang WH,Wang L,Cai M.2010.Studies on the yield,quality and virulence ratio in seeds of watermelon inoculated by Cucumber green mottle mosaic virus.Plant Protection,36(6):82-86(in Chinese)[李立梅,吴元华,赵秀香,王文航,王林,蔡明.2010.黄瓜绿斑驳花叶病毒对西瓜产量、品质及种子带毒的影响.植物保护,36(6):82-86]
Li W,Shao YZ,Chen WX.2005.Research advances in the relationships of sucrose phosphate synthase(SPS)with fruit quality and maturity and senescence.Acta Botanica Boreali-Occidentalia Sinica,25(11):2340-2344(in Chinese)[李雯,邵远志,陈维信.2005.蔗糖磷酸合成酶(SPS)与果实品质及成熟衰老的研究进展.西北植物学报,25(11):2340-2344]
Lin Y,Yang YB,Wang C,Wang WH,Mao LR.2012.A preliminary report on Cucumber green mottle mosaic virus in watermelon of Wentai area.Journal of Zhejiang Agricultural Sciences,(1):83-85(in Chinese)[林燚,杨瑜斌,王驰,王文华,毛玲荣.2012.温台地区西瓜发生黄瓜绿斑驳花叶病毒调查初报.浙江农业科学,(1):83-85]
Liu HY,Zhu ZJ,Qian QQ,Ge ZP.2004.The effects of different rootstocks on the sugar metabolism and related enzyme activities in small and early-maturing watermelon during fruit development.Acta Horticulturae Sinica,31(l):47-52(in Chinese)[刘慧英,朱祝军,钱琼秋,葛志平.2004.砧木对小型早熟西瓜果实糖代谢及相关酶活性的影响.园艺学报,31(l):47-52]
Liu Y,Peng B,Yang XH,Li XD,Gu QS.2012.Detection of Cucumber green mottle mosaic virus of watermelon blood flesh in Jinan area.China Vegetables,(10):75-79(in Chinese)[刘颖,彭斌,杨晓红,李向东,古勤生.2012.济南地区倒瓤西瓜CGMMV的检测.中国蔬菜,(10):75-79]
Park GS.2001.Occurrence of two tobamovirus diseases in cucurbits and control measures in Korea.The Plant pathology Journal,17(5):243-248
Qian QQ,Liu HY,Zhu ZJ.2004.Studies on sugar metabolism and related enzymes activity during watermelon fruit development as influenced by grafting.Journal of Zhejiang University(Agriculture and Life Sciences),30(3):285-289(in Chinese)[钱琼秋,刘慧英,朱祝军.2004.嫁接西瓜果实发育过程中蔗糖代谢及相关酶调控的研究.浙江大学学报(农业与生命科学版),30(3):285-289]
Ruan YL,Chourey PS,Delmer DP,Perez-Grau L.1997.The differential expression of sucrose synthase in relation to diverse patterns of carbon partitioning in developing cotton seeds.Plant Physiology,115(2):375-385
Schaffer AA,Petreikov M.1997.Inhibition of fructokinase and sucrose synthase by cytosolic levels of fructose in young tomato fruit undergoing transient starch synthesis.Physiologia Plantarum,101(4):800-806
Srivastava AK,Tiwari CB.1998.Polyphenoloxidase activity as influenced by CGMMV infected cucumber.Journal of Living World,5(1):7-10
Stadler R,Truermit E,Gahrtz M,Sauer N.1999.The At SUC1 sucrose carrier may represent the osmotic driving force for anther dehiscence and pollen tube growth in Arabidopsis.The Plant Journal,19(3):269-278
Tian YL,Liu DM,Zhang YJ,Li MF,Ma ZH.2009.Bioassay and genomic studies on the two isolates of Cucumber green mottle mosaic virus from Beijing and Shandong.Plant Quarantine,23(6):1-6(in Chinese)[田永蕾,刘冬梅,张永江,李明福,马占鸿.2009.黄瓜绿斑驳花叶病毒北京和山东分离物的生物学测定及其基因组比较.植物检疫,23(6):1-6]
Ugaki M,Tomiyama M,Kakutani T,Hidaka S,Kiguchi T,Nagata R,Sato T,Motoyoshi F,Nishiguchi M.1991.The complete nucleotide sequence of Cucumber green mottle mosaic virus(SH strain)genomic RNA.Journal of General Virology,72(7):1487-1495
Vani S,Varma A.1993.Properties of Cucumber green mottle mosaic virus isolated from water of river Jamuna.Indian Phytopathology,46(2):118-122
Wang YZ,Zhang DP.2000.Regulating effects of ethylene on carbohydrate metabolism in‘Starkrimson’apple fruit during the ripening period.Acta Horticulturae Sinica,27(6):391-395(in Chinese)[王永章,张大鹏.2000.乙烯对成熟期新红星苹果果实碳水化合物代谢的调控.园艺学报,27(6):391-395]
Xue YL.1985.Laboratory manual of plant physiology.Shanghai:Shanghai Science and Technology Press(in Chinese)[薛应龙.1985.植物生理学实验手册.上海:上海科学技术出版社]
Yakushiji H,Nonami H,Fukuyama T,Ono S,Takagi N,Hashimoto Y.1996.Sugar accumulation enhanced by osmoregulation in Satsuma mandarin fruit.Journal of the American Society for Horticultural Science,121:466-472
Zhou LL,Wu YH,Zhao XX,Li LM,Cai M,Wang L,Wang WH.2008.The biological characteristics of Cucumber green mottle mosaic virus and its effects on yield and quality of watermelon.Journal of Shenyang Agricultural University,39(4):417-422(in Chinese)[周玲玲,吴元华,赵秀香李立梅,蔡明,王林,王文航.2008.黄瓜绿斑驳花叶病毒生物学特性及对西瓜生长的影响.沈阳农业大学学报,39(4):417-422]