氯吡苯脲处理对莲子细胞壁多糖降解特性的影响
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摘要
为探究氯吡苯脲(1-(2-chloropyridin-4-yl)-3-phenylurea,CPPU)处理对莲子采后细胞壁多糖降解特性的影响,以‘太空莲36号’莲蓬为试材,以清水浸泡为对照,以5 mg/L CPPU对莲蓬进行处理。结果表明:CPPU处理可有效维持莲蓬及莲子较好的表型,保持莲子脆嫩口感;CPPU处理的莲子含有更高的共价结合果胶含量和更完整的纤维素多糖碳链结构;CPPU处理可有效抑制果胶甲酯酶、多聚半乳糖醛酸酶等果胶降解酶和外切葡聚糖酶、β-葡萄糖苷酶、内切葡聚糖酶等纤维素降解酶的活力,从而延缓共价结合果胶降解及水溶性果胶、离子结合果胶积累,并保持莲子纤维素和半纤维素含量;同时,CPPU处理保持了莲子细胞壁结构和纤维素微纤丝的形态,并抑制了细胞的质壁分离。因此,CPPU处理有利于抑制莲子细胞壁多糖的降解,更好地维持细胞完整性,保持莲子的口感,从而延缓其采后衰老。本实验为鲜莲蓬和莲子的采后保鲜提供理论依据和技术支持。
In order to investigate the effect of 1-(2-chloropyridin-4-yl)-3-phenylurea(CPPU) treatment on the degradation of cell wall polysaccharides in lotus seeds, lotus pods from the cultivar ‘Taikonglian 36' were treated with 5 mg/L CPPU. The samples treated with distilled water were used as controls. It was found that CPPU treatment could significantly maintain good phenotype of lotus pods and seeds, as well as crisp and tender taste of the seeds as compared with control. Meanwhile, CPPU treatment increased covalently bound pectin content in lotus seeds and maintained the structural integrity of cellulose polysaccharide chains. In addition, CPPU treatment inhibited the activity of pectin-degrading enzymes such as pectin methylesterase(PME) and polygalacturonase(PG) and the activity of cellulose-degrading enzymes such as cellobiohydrolase 1(C1), β-1,4-endoglucanase(Cx) and β-glucosidase in lotus seeds, thus degrading the degradation of covalently bound pectin and the accumulation of water-soluble pectin and ion-bound pectin and maintaining cellulose and hemicellulose. CPPU treatment could maintain the integrity of the cell wall and the morphology of cellulose microfibrils in lotus seeds and inhibited plasmolysis. Therefore, CPPU treatment can inhibit the degradation of cell wall polysaccharides, maintain cell integrity in lotus seeds as well as the taste and delay postharvest senescence of lotus pods and seeds. This experiment can provide theoretical and technical support for postharvest preservation of lotus pods and seeds.
In order to investigate the effect of 1-(2-chloropyridin-4-yl)-3-phenylurea(CPPU) treatment on the degradation of cell wall polysaccharides in lotus seeds, lotus pods from the cultivar ‘Taikonglian 36' were treated with 5 mg/L CPPU. The samples treated with distilled water were used as controls. It was found that CPPU treatment could significantly maintain good phenotype of lotus pods and seeds, as well as crisp and tender taste of the seeds as compared with control. Meanwhile, CPPU treatment increased covalently bound pectin content in lotus seeds and maintained the structural integrity of cellulose polysaccharide chains. In addition, CPPU treatment inhibited the activity of pectin-degrading enzymes such as pectin methylesterase(PME) and polygalacturonase(PG) and the activity of cellulose-degrading enzymes such as cellobiohydrolase 1(C1), β-1,4-endoglucanase(Cx) and β-glucosidase in lotus seeds, thus degrading the degradation of covalently bound pectin and the accumulation of water-soluble pectin and ion-bound pectin and maintaining cellulose and hemicellulose. CPPU treatment could maintain the integrity of the cell wall and the morphology of cellulose microfibrils in lotus seeds and inhibited plasmolysis. Therefore, CPPU treatment can inhibit the degradation of cell wall polysaccharides, maintain cell integrity in lotus seeds as well as the taste and delay postharvest senescence of lotus pods and seeds. This experiment can provide theoretical and technical support for postharvest preservation of lotus pods and seeds.
引文
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[9] LUO Shufen, HU Huali, ZHANG Leigang, et al. Sugars in postharvest lotus seeds were modified by 6-benzylaminopurine treatment through altering related enzymes involved in starch-sucrose metabolism[J]. Scientia Horticulturae, 2017, 221: 73-82. DOI:10.1016/j.scienta.2017.03.044.
[10] CRUZ-CASTILLO J G, BALDICCHI A, FRIONI T, et al. Preanthesis CPPU low dosage application increases ‘Hayward’ kiwifruit weight without affecting the other qualitative and nutritional characteristics[J]. Food Chemistry, 2014, 158: 224-228. DOI:10.1016/j.foodchem.2014.01.131.
[11]陈巧玲,叶蕙,刘伟,等.C P P U延缓菜心叶片衰老的作用[J].西北植物学报, 2001, 21(6): 1245-1248. DOI:10.3321/j.issn:1000-4025.2001.06.032.
[12]梁颖.KT-30对离体水稻叶片衰老进程的影响[J].西南农业学报,2002, 15(3): 53-56. DOI:10.3969/j.issn.1001-4829.2002.03.012.
[13]张鲁斌,宋康华,贾志伟,等.氯吡脲(CPPU)处理对采后杧果保鲜及生理效应的影响[J].果树学报, 2016, 33(5): 604-611. DOI:10.13925/j.cnki.gsxb.20150388.
[14] HUANG Hua, JIANG Yueming. Effect of plant growth regulators on banana fruit and broccoli during storage[J]. Scientia Horticulturae, 2012, 145: 62-67. DOI:10.1016/j.scienta.2012.07.025.
[15]张美芳,何玲,张美丽,等.猕猴桃鲜果贮藏保鲜研究进展[J].食品科学, 2014, 35(11): 343-347. DOI:10.7506/spkx1002-6630-201411066.
[16]周宏胜,孙凤杰,罗淑芬,等.氯吡苯脲处理对采后莲蓬保鲜效果的影响[J].食品科学,2018, 39(15): 225-232. DOI:10.7506/spkx1002-6630-201815033.
[17]茅林春,张上隆.间歇低温胁迫对桃果实细胞壁代谢的影响[J].植物生理学报, 2 0 0 1, 2 7(2): 1 5 1-1 5 5. D O I:1 0.3 3 2 1/j.issn:1671-3877.2001.02.010.
[18] MCCANN M C, SHI J, ROBERS K, et al. Changes in pectin structure and localization during the growth of unadapted and NaCl-adapted tobacco cells[J]. The Plant Journal, 1994, 5(6): 773-785. DOI:10.1046/j.1365-313X.1994.5060773.x.
[19] CARVAJAL F, PALMA F, JAMILENA M, et al. Cell wall metabolism and chilling injury during postharvest cold storage in zucchini fruit[J]. Postharvest Biology and Technology, 2015, 108: 68-77. DOI:10.1016/j.postharvbio.2015.05.013.
[20]李合生.植物生理生化实验原理和技术[M].北京: 高等教育出版社,2000: 211-212.
[21]辜青青,唐红英,魏清江,等.南丰蜜橘果实纤维素代谢与化渣的关系研究[J].园艺学报, 2016, 43(5): 867-875. DOI:10.16420/j.issn.0513-353x.2015-0750.
[22]吴雪莹,屈立武,周雅涵,等.壳聚糖和纳米SiOx处理对采后脐橙果实硬度的影响[J].食品科学, 2015, 36(2): 204-209. DOI:10.7506/spkx1002-6630-201502040.
[23] KHALIQ G, MOHAMED M T M, ALI A, et al. Effect of gum arabic coating combined with calcium chloride on physico-chemical and qualitative properties of mango (Mangifera indica L.) fruit during low temperature storage[J]. Scientia Horticulturae, 2015, 190: 187-194. DOI:10.1016/j.scienta.2015.04.020.
[24] SZYMA?SKA-CHARGOT M, CHYLINSKA M, KRUK B, et al. Combining FT-IR spectroscopy and multivariate analysis for qualitative and quantitative analysis of the cell wall composition changes during apples development[J]. Carbohydrate Polymers, 2015, 115: 93-103. DOI:10.1016/j.carbpol.2014.08.039.
[25] SZYMANSKA-CHARGOT M, CYBULSKA J, ZDUNEK A. Sensing the structural differences in cellulose from apple and bacterial cell wall materials by Raman and FT-IR spectroscopy[J]. Sensors, 2011, 11(6): 5543-5560. DOI:10.3390/s110605543.
[26] ZHBANKOV R G, FIRSOV S P, BUSLOV D K, et al. Structural physico-chemistry of cellulose macromolecules: vibrational spectra and structure of cellulose[J]. Journal of Molecular Structure, 2002, 614(1/2/3): 117-125. DOI:10.1016/S0022-2860(02)00252-1.
[27] ABDENOR F, PADMESH A, MARKR W, et al. Determining the degree of methylesterification of pectin by ATR/FT-IR: methodology optimisation and comparison with theoretical calculations[J]. Carbohydrate Polymers, 2009, 78(4): 847-853. DOI:10.1016/j.carbpol.2009.07.003.
[28]王梦,段德超,徐辰,等.茶树根细胞壁不同组分对铅的吸附性能及其功能团的傅里叶红外光谱学研究[J].生态学报, 2015, 35(6): 1743-1751. DOI:10.5846/stxb201305291222.
[29] GOULAO L F, OLIVEIRA C M. Cell wall modifications during fruit ripening: when a fruit is not the fruit[J]. Trends in Food Science & Technology, 2008, 19(1): 4-25. DOI:10.1016/j.tifs.2007.07.002.
[30] SáNCHEZ C. Lignocellulosic residues: biodegradation and bioconversion by fungi[J]. Biotechnology Advances, 2009, 27(2): 185-194. DOI:10.1007/s00158-003-0325-4.
[31] SEYMOUR G B, GROSS K C. Cell wall disassembly and fruit softening[J]. Postharvest News and Information, 1996, 7(3): 45-52.
[32]罗淑芬,孙凤杰,胡花丽,等.6-苄基腺嘌呤对鲜莲子活性氧代谢及能量水平的影响[J].核农学报, 2018, 32(5): 916-923. DOI:10.11869/j.issn.100-8551.2018.05.0916.
[33]刘莹,马慧,刘艳,等.机械损伤对甜瓜果实贮藏期间膜脂过氧化和细胞壁代谢的影响[C]//中国园艺学会2015年学术年会论文摘要集.厦门: 园艺学报编辑部, 2015: 202.
[34]王三根.细胞分裂素在植物抗逆和延衰中的作用[J].植物学通报, 2 0 0 0, 1 7(2): 1 2 1-1 2 6; 1 6 7. D O I:1 0.3 9 6 9/j.issn.1674-3466.2000.02.004.
[35]龚吉军,谭兴和,夏延斌,等.小袋包装气调和6-BA对黄花菜采后生理变化的影响[J].食品科学, 2004, 25(6): 175-178. DOI:10.3321/j.issn:1002-6630.2004.06.046.
[2]郑天闻,徐婷婷,范亚苇,等.莲子多酚氧化酶的酶学性质[J].食品工业科技, 2014, 35(2): 162-165; 170. DOI:10.13386/j.issn1002-0306.2014.02.039.
[3]王凌云,杨梦飞,郑寨生,等.不同温度条件对鲜食莲子品质的影响[J].现代农业科技, 2013(23): 296; 299. DOI:10.3969/j.issn.1007-5739.2013.23.186.
[4] LIU S Y, QIU F, KONG W J, et al. Development and validation of an accurate and rapid LC-ESI-MS/MS method for the simultaneous quantification of aflatoxin B1,B2,G1 and G2 in lotus seeds[J]. Food Control, 2013, 29(1): 156-161. DOI:10.1016/j.foodcont.2012.05.069.
[5]高建晓,王毓宁,李鹏霞,等.漆蜡涂膜对鲜莲蓬采后褐变的影响[J].食品科学, 2016, 37(18): 275-282. DOI:10.7506/spkx1002-6630-201618044.
[6]王建辉,靳娜,刘永乐,等.低温处理对湘莲采后生理变化的影响[J].食品科学, 2014, 35(18): 209-213. DOI:10.7506/spkx1002-6630-201418040.
[7]朱雁青,胡花丽,郭峰,等.不同薄膜包装对鲜莲蓬褐变的影响[J].食品与发酵工业, 2014, 40(10): 97-102.
[8] LI P X, HU H L, LUO S F, et al. Shelf life extension of fresh lotus pods and seeds (Nelumbo nucifera, Gaertn.) in response to treatments with 1-MCP and lacquer wax[J]. Postharvest Biology and Technology, 2017, 125: 140-149. DOI:10.1016/j.postharvbio.2016.10.004.
[9] LUO Shufen, HU Huali, ZHANG Leigang, et al. Sugars in postharvest lotus seeds were modified by 6-benzylaminopurine treatment through altering related enzymes involved in starch-sucrose metabolism[J]. Scientia Horticulturae, 2017, 221: 73-82. DOI:10.1016/j.scienta.2017.03.044.
[10] CRUZ-CASTILLO J G, BALDICCHI A, FRIONI T, et al. Preanthesis CPPU low dosage application increases ‘Hayward’ kiwifruit weight without affecting the other qualitative and nutritional characteristics[J]. Food Chemistry, 2014, 158: 224-228. DOI:10.1016/j.foodchem.2014.01.131.
[11]陈巧玲,叶蕙,刘伟,等.C P P U延缓菜心叶片衰老的作用[J].西北植物学报, 2001, 21(6): 1245-1248. DOI:10.3321/j.issn:1000-4025.2001.06.032.
[12]梁颖.KT-30对离体水稻叶片衰老进程的影响[J].西南农业学报,2002, 15(3): 53-56. DOI:10.3969/j.issn.1001-4829.2002.03.012.
[13]张鲁斌,宋康华,贾志伟,等.氯吡脲(CPPU)处理对采后杧果保鲜及生理效应的影响[J].果树学报, 2016, 33(5): 604-611. DOI:10.13925/j.cnki.gsxb.20150388.
[14] HUANG Hua, JIANG Yueming. Effect of plant growth regulators on banana fruit and broccoli during storage[J]. Scientia Horticulturae, 2012, 145: 62-67. DOI:10.1016/j.scienta.2012.07.025.
[15]张美芳,何玲,张美丽,等.猕猴桃鲜果贮藏保鲜研究进展[J].食品科学, 2014, 35(11): 343-347. DOI:10.7506/spkx1002-6630-201411066.
[16]周宏胜,孙凤杰,罗淑芬,等.氯吡苯脲处理对采后莲蓬保鲜效果的影响[J].食品科学,2018, 39(15): 225-232. DOI:10.7506/spkx1002-6630-201815033.
[17]茅林春,张上隆.间歇低温胁迫对桃果实细胞壁代谢的影响[J].植物生理学报, 2 0 0 1, 2 7(2): 1 5 1-1 5 5. D O I:1 0.3 3 2 1/j.issn:1671-3877.2001.02.010.
[18] MCCANN M C, SHI J, ROBERS K, et al. Changes in pectin structure and localization during the growth of unadapted and NaCl-adapted tobacco cells[J]. The Plant Journal, 1994, 5(6): 773-785. DOI:10.1046/j.1365-313X.1994.5060773.x.
[19] CARVAJAL F, PALMA F, JAMILENA M, et al. Cell wall metabolism and chilling injury during postharvest cold storage in zucchini fruit[J]. Postharvest Biology and Technology, 2015, 108: 68-77. DOI:10.1016/j.postharvbio.2015.05.013.
[20]李合生.植物生理生化实验原理和技术[M].北京: 高等教育出版社,2000: 211-212.
[21]辜青青,唐红英,魏清江,等.南丰蜜橘果实纤维素代谢与化渣的关系研究[J].园艺学报, 2016, 43(5): 867-875. DOI:10.16420/j.issn.0513-353x.2015-0750.
[22]吴雪莹,屈立武,周雅涵,等.壳聚糖和纳米SiOx处理对采后脐橙果实硬度的影响[J].食品科学, 2015, 36(2): 204-209. DOI:10.7506/spkx1002-6630-201502040.
[23] KHALIQ G, MOHAMED M T M, ALI A, et al. Effect of gum arabic coating combined with calcium chloride on physico-chemical and qualitative properties of mango (Mangifera indica L.) fruit during low temperature storage[J]. Scientia Horticulturae, 2015, 190: 187-194. DOI:10.1016/j.scienta.2015.04.020.
[24] SZYMA?SKA-CHARGOT M, CHYLINSKA M, KRUK B, et al. Combining FT-IR spectroscopy and multivariate analysis for qualitative and quantitative analysis of the cell wall composition changes during apples development[J]. Carbohydrate Polymers, 2015, 115: 93-103. DOI:10.1016/j.carbpol.2014.08.039.
[25] SZYMANSKA-CHARGOT M, CYBULSKA J, ZDUNEK A. Sensing the structural differences in cellulose from apple and bacterial cell wall materials by Raman and FT-IR spectroscopy[J]. Sensors, 2011, 11(6): 5543-5560. DOI:10.3390/s110605543.
[26] ZHBANKOV R G, FIRSOV S P, BUSLOV D K, et al. Structural physico-chemistry of cellulose macromolecules: vibrational spectra and structure of cellulose[J]. Journal of Molecular Structure, 2002, 614(1/2/3): 117-125. DOI:10.1016/S0022-2860(02)00252-1.
[27] ABDENOR F, PADMESH A, MARKR W, et al. Determining the degree of methylesterification of pectin by ATR/FT-IR: methodology optimisation and comparison with theoretical calculations[J]. Carbohydrate Polymers, 2009, 78(4): 847-853. DOI:10.1016/j.carbpol.2009.07.003.
[28]王梦,段德超,徐辰,等.茶树根细胞壁不同组分对铅的吸附性能及其功能团的傅里叶红外光谱学研究[J].生态学报, 2015, 35(6): 1743-1751. DOI:10.5846/stxb201305291222.
[29] GOULAO L F, OLIVEIRA C M. Cell wall modifications during fruit ripening: when a fruit is not the fruit[J]. Trends in Food Science & Technology, 2008, 19(1): 4-25. DOI:10.1016/j.tifs.2007.07.002.
[30] SáNCHEZ C. Lignocellulosic residues: biodegradation and bioconversion by fungi[J]. Biotechnology Advances, 2009, 27(2): 185-194. DOI:10.1007/s00158-003-0325-4.
[31] SEYMOUR G B, GROSS K C. Cell wall disassembly and fruit softening[J]. Postharvest News and Information, 1996, 7(3): 45-52.
[32]罗淑芬,孙凤杰,胡花丽,等.6-苄基腺嘌呤对鲜莲子活性氧代谢及能量水平的影响[J].核农学报, 2018, 32(5): 916-923. DOI:10.11869/j.issn.100-8551.2018.05.0916.
[33]刘莹,马慧,刘艳,等.机械损伤对甜瓜果实贮藏期间膜脂过氧化和细胞壁代谢的影响[C]//中国园艺学会2015年学术年会论文摘要集.厦门: 园艺学报编辑部, 2015: 202.
[34]王三根.细胞分裂素在植物抗逆和延衰中的作用[J].植物学通报, 2 0 0 0, 1 7(2): 1 2 1-1 2 6; 1 6 7. D O I:1 0.3 9 6 9/j.issn.1674-3466.2000.02.004.
[35]龚吉军,谭兴和,夏延斌,等.小袋包装气调和6-BA对黄花菜采后生理变化的影响[J].食品科学, 2004, 25(6): 175-178. DOI:10.3321/j.issn:1002-6630.2004.06.046.