脱硫对熏硫‘黑叶’荔枝采后品质及果肉亚硫酸盐代谢的影响
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摘要
本实验研究了脱硫对熏硫‘黑叶’荔枝果实采后品质、亚硫酸盐残留量和果肉中亚硫酸盐代谢的影响。测定对照(施保克)、熏硫和脱硫处理的果实4℃贮藏期间的色度值、褐变指数、腐烂率、亚硫酸盐残留量及果肉中亚硫酸盐氧化酶(sul?te oxidase,SO)、腺苷-5’-磷酰硫酸还原酶(adenosine 5’-phosphosulfatezoline reductase,APR)、亚硫酸盐还原酶(sulfite reductase,SiR)、丝氨酸酰基转移酶(serine acetyltransferase,SAT)和乙酰丝氨酸裂解酶(O-acetylserine (thiol) lyase,OAS-TL)活力和基因表达。结果表明:贮藏期间脱硫果实较熏硫果实提前转红;脱硫和熏硫荔枝的果皮褐变指数和腐烂率均较对照组显著降低;相比熏硫,脱硫使果皮水分质量分数降低和相对电导率升高的幅度更大,抑制呼吸的效果则相近;脱硫使果皮亚硫酸盐含量大幅降低,并使得果肉亚硫酸盐含量在贮藏16 d后降至接近对照组的水平;贮藏期间除APR外,熏硫和脱硫后果肉中其他4种酶的活力均呈现上调,即贮藏过程中处理组SO、SAT活力分别较同时期对照组上调2.0、8.6～26.7倍,而脱硫果肉SAT活力仅在贮藏0 d较对照组上调11.1倍,之后迅速下降,贮藏32 d后与对照组无显著差异;熏硫和脱硫果肉中5个基因的表达则均在贮藏中后期总体高于对照。上述结果表明,脱硫可加快熏硫荔枝果皮恢复红色,获得与熏硫相当的保鲜效果,大幅降低了熏硫荔枝的硫残留,保障其食用安全。脱硫和熏硫荔枝果肉中SO和SAT活力的上调表明,SO酶促氧化反应是荔枝果肉亚硫酸盐降解中占主导和最稳定的方式,SAT和SiR主导的还原途径则可能作为氧化途径的补充。
The effect of desulfurization on the postharvest quality, sulfite residue and sulfite degradation of sulfurfumigated ‘Heiye' litchi fruit was investigated. The color, browning index, decay incidence, sul?te residue of litchi fruit,and the activities and gene expression of sul?te oxidase(SO), adenosine 5'-phosphosulfatezoline reductase(APR), sul?te reductase(SiR), serine acetyltransferase(SAT) and O-acetylserine(thiol) lyase(OAS-TL) in pulp were measured in the control(treated with prochloraz), sulfur-fumigated and desulfurized fruits during storage at 4 ℃. Our results showed that the desulfurized fruit turned red earlier than the sulfur-fumigated fruit during storage. The browning index and decay incidence of the treated fruits were signi?cantly lower than those of the control fruit. Desulfurized fruit showed a greater reduction in the moisture content and a greater increase in the relative conductivity of pericarp than did sulfur-fumigated fruit, whereas both treatments were similarly effective against respiration. After desulphurization, sul?te content in pericarp decreased signi?cantly, and sul?te content in desulfurized litchi pulp decreased to a level comparable to that of the control on the 16 th day of storage. During storage, the activities of all enzymes investigated except for APR increased in sulfur-fumigated and desulfurized fruits when compared with the control; the activities of SO and SAT in the treated fruits increased by 2.0 and8.6–26.7 times, respectively. On day 0 of storage, SAT activity in desulfurized litchi increased by 11.1 times in comparison with the control, and then it decreased rapidly, reaching a level not signi?cantly different from that of the control on Day 32.The gene expression of all ?ve enzymes in sulfur-fumigated and desulfurized fruits was higher than that in the control at the middle and late stages of storage. This study indicated that desulfurization accelerated color recovery in the pericarp of sulfur-fumigated litchi fruit with achieving a preservation effect comparable to sulphur-fumigation, and dramatically reduced sul?te residue, thereby ensuring its safety for consumption. The increasing activity of SO and SAT in sulfur-fumigated and desulfurized fruit pulp indicated that oxidative degradation of sul?te catalyzed by SO was dominant and was the most stable way, which may be complemented by the reduction pathway controlled by SAT and SiR.
The effect of desulfurization on the postharvest quality, sulfite residue and sulfite degradation of sulfurfumigated ‘Heiye' litchi fruit was investigated. The color, browning index, decay incidence, sul?te residue of litchi fruit,and the activities and gene expression of sul?te oxidase(SO), adenosine 5'-phosphosulfatezoline reductase(APR), sul?te reductase(SiR), serine acetyltransferase(SAT) and O-acetylserine(thiol) lyase(OAS-TL) in pulp were measured in the control(treated with prochloraz), sulfur-fumigated and desulfurized fruits during storage at 4 ℃. Our results showed that the desulfurized fruit turned red earlier than the sulfur-fumigated fruit during storage. The browning index and decay incidence of the treated fruits were signi?cantly lower than those of the control fruit. Desulfurized fruit showed a greater reduction in the moisture content and a greater increase in the relative conductivity of pericarp than did sulfur-fumigated fruit, whereas both treatments were similarly effective against respiration. After desulphurization, sul?te content in pericarp decreased signi?cantly, and sul?te content in desulfurized litchi pulp decreased to a level comparable to that of the control on the 16 th day of storage. During storage, the activities of all enzymes investigated except for APR increased in sulfur-fumigated and desulfurized fruits when compared with the control; the activities of SO and SAT in the treated fruits increased by 2.0 and8.6–26.7 times, respectively. On day 0 of storage, SAT activity in desulfurized litchi increased by 11.1 times in comparison with the control, and then it decreased rapidly, reaching a level not signi?cantly different from that of the control on Day 32.The gene expression of all ?ve enzymes in sulfur-fumigated and desulfurized fruits was higher than that in the control at the middle and late stages of storage. This study indicated that desulfurization accelerated color recovery in the pericarp of sulfur-fumigated litchi fruit with achieving a preservation effect comparable to sulphur-fumigation, and dramatically reduced sul?te residue, thereby ensuring its safety for consumption. The increasing activity of SO and SAT in sulfur-fumigated and desulfurized fruit pulp indicated that oxidative degradation of sul?te catalyzed by SO was dominant and was the most stable way, which may be complemented by the reduction pathway controlled by SAT and SiR.
引文
[1]舒肇甦.我国荔枝出口现状与发展建议[J].保鲜与加工,2006(6):1-4.
[2]王丽丽,纪淑娟,李顺.食品中二氧化硫及亚硫酸盐的作用与检测方法[J].食品与药品,2007,9(2):64-66.
[3]梁骏,王杰,张库凌.出口保鲜荔枝二氧化硫残留量变化分析[J].中国热带农业,2008(1):46-47.
[4]韩冬梅,吴振先,季作梁,等.SO_2对龙眼果实的氧化作用与衰老的影响[J].果树科学,1999,16(1):24-29.
[5]吴振先,韩冬梅,季作梁,等.SO_2对贮藏龙眼果皮酶促褐变的影响[J].园艺学报,1999,26(2):91-95.
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[7]赵秀娟,李益先,麦任娣.浅析荔枝的保鲜及出口[J].福建果树,2007(1):34-35.
[8]HEBER U,HüVE K.Action of SO_2 on plants and metabolic detoxification of SO_2[J].International Review of Cytology,1997,177:255-286.DOI:10.1016/S0074-7696(08)62234-2.
[9]HAMISCH D,RANDEWIG D,SCHLIESKY S,et al.Impact of SO_2 on Arabidopsis thaliana transcriptome in wildtype and sulfite oxidase knockout plants analyzed by RNA deep sequencing[J].New Phytologist,2012,196(4):1074-1085.DOI:10.1111/j.1469-8137.2012.04331.x.
[10]BAILLIE C K,KAUFHOLDT D,KARPINSKI L H,et al.Detoxification of volcanic sulfur surplus in planta:three different strategies of survival[J].Environmental and Experimental Botany,2016,126:44-54.DOI:10.1016/j.envexpbot.2016.02.007.
[11]AGHAJANZADEH T,HAWKESFORD M J,DE KOK L J.Atmospheric H2S and SO_2 as sulfur sources for Brassica juncea and Brassica rapa:regulation of sulfur uptake and assimilation[J].Environmental and Experimental Botany,2016,124:1-10.DOI:10.1016/j.envexpbot.2015.12.001.
[12]CHAO D Y,BARANIECKA P,DANKU J,et al.Variation in sulfur and selenium accumulation is controlled by naturally occurring isoforms of the key sulfur assimilation enzyme adenosine 5’-phosphosulfate reductase 2 across the arabidopsis species range[J].Plant Physiology,2014,166(3):1593-1608.DOI:10.1104/pp.114.247825.
[13]BRYCHKOVA G,YARMOLINSKY D,FLUHR R,et al.The determination of sulfite levels and its oxidation in plant leaves[J].Plant Science,2012,190(8):123-130.DOI:10.1016/j.plantsci.2012.04.004.
[14]OSTROWSKI J,WU J Y,RUEGER D C,et al.Characterization of the cysJIH regions of Salmonella typhimurium and Escherichia coli B.DNA sequences of cysI and cysH and a model for the siroheme-Fe4S4active center of sulfite reductase hemoprotein based on amino acid homology with spinach nitrite reductase[J].The Journal of Biological Chemistry,1989,264(26):15726-15737.
[15]RANDEWIG D,HAMISCH D,EIBLMEIER M,et al.Oxidation and reduction of sulfite contribute to susceptibility and detoxification of SO_2 in Populus×canescens leaves[J].Trees,2014,28:399-411.DOI:10.1007/s00468-013-0958-x.
[16]CHRONIS D,KRISHNAN H B.Sulfur assimilation in soybean:molecular cloning and characterization of O-acetylserine(thiol)lyase(cysteine synthase)[J].Crop Science,2003,43(5):1819-1827.DOI:10.2135/cropsci2003.1819.
[17]H?NSCH R,LANG C,RIEBESEEL E,et al.Plant sulfite oxidase as novel producer of H_2O_2:combination of enzyme catalysis with a subsequent non-enzymatic reaction step[J].Journal of Biological Chemistry,2006,281(10):6884-6888.DOI:10.1074/jbc.M513054200.
[18]夏宗良,武轲,王永霞,等.玉米亚硫酸氧化酶基因ZmSO的克隆及在二氧化硫胁迫下的表达分析[J].中国农业科学,2009,42(8):2643-2651.
[19]XIA Zongliang,SUN Kaile,WANG Meiping,et al.Overexpression of a maize sulfite oxidase gene in tobacco enhances tolerance to sulfite stress via sulfite oxidation and CAT-mediated H_2O_2 scavenging[J].PLoS ONE,2012,7(5):e37383.DOI:10.1371/journal.pone.0037383.
[20]SCHEERER U,HAENSCH R,MENDEL R R,et al.Sulphur flux through the sulphate assimilation pathway is differently controlled by adenosine 5’-phosphosulphate reductase under stress and in transgenic poplar plants overexpressingγ-ECS,SO,or APR[J].Journal of Experimental Botany,2010,61(2):609-622.DOI:10.1093/jxb/erp327.
[21]吴振先.荔枝采后贮藏褐变过程中的一些生理生化变化[D].广州:华南农业大学,1995:28.
[22]陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2002:115-116.
[23]季作梁,张昭其,王燕,等.芒果低温贮藏及其冷害的研究[J].园艺学报,1994,21(2):111-116.
[24]SCHMITTGEN T D,LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocol,2008,3:1101-1108.DOI:10.1038/nprot.2008.73.
[25]美国国家档案,登录管理局联邦注册处.美国联邦管理法规.FDA食品法规(2001版)[M].中国轻工业上海设计院,译.北京:中国轻工业出版社,2003:100-199.
[26]H?NSCH R,LANG C,RENNENBERG H,et al.Significance of plant sulfite oxidase[J].Plant Biology,2007,9(5):589-595.DOI:10.1055/s-2007-965433.
[27]KAUFHOLDT D,BAILLIE C K,WILLE T,et al.Prospective posttranslational regulation of plant sulfite oxidase[M]//DE KOK L J,HAWKESFORD M J,RENNENBERG H,et al.Molecular physiology and ecophysiology of sulfur.Switzerland:Springer International Publishing,2015:179-187.DOI:10.1007/978-3-319-20137-5_18.
[28]汪伟,张伟,朱丽琴,等.植物硫化氢生理效应及机制研究进展[J].中国农学通报,2013,29(31):78-82.
[29]ZHANG H,HU L Y,HU K D,et al.Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress[J].Journal of Integrative Plant Biology,2008,50(12):1518-1529.DOI:10.1111/j.1744-7909.2008.00769.x.
[30]GARCíA-MATA C,LAMATTINA L.Hydrogen sulphide,a novel gasotransmitter involved in guard cell signalling[J].New Phytologist,2010,188(4):977-984.DOI:10.1111/j.1469-8137.2010.03465.x.
[31]SCUFFI D,áLVAREZ C,LASPINA N,et al.Hydrogen sulfide generated by L-cysteine desulfhydrase acts upstream of nitric oxide to modulate abscisic acid-dependent stomatal closure[J].Plant Physiology,2014,166(4):2065-2076.DOI:10.1104/pp.114.245373.
[32]HOU Z H,WANG L X,LIU J,et al.Hydrogen sulfide regulates ethylene-induced stomatal closure in Arabidopsis thaliana[J].Journal of Integrative Plant Biology,2013,55(3):277-289.DOI:10.1111/jipb.12004.
[33]ZHANG H,HU S L,ZHANG Z J,et al.Hydrogen sulfide acts as a regulator of flower senescence in plants[J].Postharvest Biology and Technology,2011,60:251-257.DOI:10.1016/j.postharvbio.2011.01.006.
[34]HU L Y,HU S L,WU J,et al.Hydrogen sulfide prolongs postharvest shelf life of strawberry and plays an antioxidative role in fruits[J].Journal of Agriculture and Food Chemistry,2012,60(35):8684-8693.DOI:10.1021/jf300728h.
[2]王丽丽,纪淑娟,李顺.食品中二氧化硫及亚硫酸盐的作用与检测方法[J].食品与药品,2007,9(2):64-66.
[3]梁骏,王杰,张库凌.出口保鲜荔枝二氧化硫残留量变化分析[J].中国热带农业,2008(1):46-47.
[4]韩冬梅,吴振先,季作梁,等.SO_2对龙眼果实的氧化作用与衰老的影响[J].果树科学,1999,16(1):24-29.
[5]吴振先,韩冬梅,季作梁,等.SO_2对贮藏龙眼果皮酶促褐变的影响[J].园艺学报,1999,26(2):91-95.
[6]LU S L,YANG X Z,LI X H,et al.Effect of sulfur dioxide treatment on storage quality and SO_2 residue of Victoria grape[J].Advanced Materials Research,2013,798/799:1033-1036.DOI:10.4028/www.scientific.net/AMR.798-799.1033.
[7]赵秀娟,李益先,麦任娣.浅析荔枝的保鲜及出口[J].福建果树,2007(1):34-35.
[8]HEBER U,HüVE K.Action of SO_2 on plants and metabolic detoxification of SO_2[J].International Review of Cytology,1997,177:255-286.DOI:10.1016/S0074-7696(08)62234-2.
[9]HAMISCH D,RANDEWIG D,SCHLIESKY S,et al.Impact of SO_2 on Arabidopsis thaliana transcriptome in wildtype and sulfite oxidase knockout plants analyzed by RNA deep sequencing[J].New Phytologist,2012,196(4):1074-1085.DOI:10.1111/j.1469-8137.2012.04331.x.
[10]BAILLIE C K,KAUFHOLDT D,KARPINSKI L H,et al.Detoxification of volcanic sulfur surplus in planta:three different strategies of survival[J].Environmental and Experimental Botany,2016,126:44-54.DOI:10.1016/j.envexpbot.2016.02.007.
[11]AGHAJANZADEH T,HAWKESFORD M J,DE KOK L J.Atmospheric H2S and SO_2 as sulfur sources for Brassica juncea and Brassica rapa:regulation of sulfur uptake and assimilation[J].Environmental and Experimental Botany,2016,124:1-10.DOI:10.1016/j.envexpbot.2015.12.001.
[12]CHAO D Y,BARANIECKA P,DANKU J,et al.Variation in sulfur and selenium accumulation is controlled by naturally occurring isoforms of the key sulfur assimilation enzyme adenosine 5’-phosphosulfate reductase 2 across the arabidopsis species range[J].Plant Physiology,2014,166(3):1593-1608.DOI:10.1104/pp.114.247825.
[13]BRYCHKOVA G,YARMOLINSKY D,FLUHR R,et al.The determination of sulfite levels and its oxidation in plant leaves[J].Plant Science,2012,190(8):123-130.DOI:10.1016/j.plantsci.2012.04.004.
[14]OSTROWSKI J,WU J Y,RUEGER D C,et al.Characterization of the cysJIH regions of Salmonella typhimurium and Escherichia coli B.DNA sequences of cysI and cysH and a model for the siroheme-Fe4S4active center of sulfite reductase hemoprotein based on amino acid homology with spinach nitrite reductase[J].The Journal of Biological Chemistry,1989,264(26):15726-15737.
[15]RANDEWIG D,HAMISCH D,EIBLMEIER M,et al.Oxidation and reduction of sulfite contribute to susceptibility and detoxification of SO_2 in Populus×canescens leaves[J].Trees,2014,28:399-411.DOI:10.1007/s00468-013-0958-x.
[16]CHRONIS D,KRISHNAN H B.Sulfur assimilation in soybean:molecular cloning and characterization of O-acetylserine(thiol)lyase(cysteine synthase)[J].Crop Science,2003,43(5):1819-1827.DOI:10.2135/cropsci2003.1819.
[17]H?NSCH R,LANG C,RIEBESEEL E,et al.Plant sulfite oxidase as novel producer of H_2O_2:combination of enzyme catalysis with a subsequent non-enzymatic reaction step[J].Journal of Biological Chemistry,2006,281(10):6884-6888.DOI:10.1074/jbc.M513054200.
[18]夏宗良,武轲,王永霞,等.玉米亚硫酸氧化酶基因ZmSO的克隆及在二氧化硫胁迫下的表达分析[J].中国农业科学,2009,42(8):2643-2651.
[19]XIA Zongliang,SUN Kaile,WANG Meiping,et al.Overexpression of a maize sulfite oxidase gene in tobacco enhances tolerance to sulfite stress via sulfite oxidation and CAT-mediated H_2O_2 scavenging[J].PLoS ONE,2012,7(5):e37383.DOI:10.1371/journal.pone.0037383.
[20]SCHEERER U,HAENSCH R,MENDEL R R,et al.Sulphur flux through the sulphate assimilation pathway is differently controlled by adenosine 5’-phosphosulphate reductase under stress and in transgenic poplar plants overexpressingγ-ECS,SO,or APR[J].Journal of Experimental Botany,2010,61(2):609-622.DOI:10.1093/jxb/erp327.
[21]吴振先.荔枝采后贮藏褐变过程中的一些生理生化变化[D].广州:华南农业大学,1995:28.
[22]陈建勋,王晓峰.植物生理学实验指导[M].广州:华南理工大学出版社,2002:115-116.
[23]季作梁,张昭其,王燕,等.芒果低温贮藏及其冷害的研究[J].园艺学报,1994,21(2):111-116.
[24]SCHMITTGEN T D,LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J].Nature Protocol,2008,3:1101-1108.DOI:10.1038/nprot.2008.73.
[25]美国国家档案,登录管理局联邦注册处.美国联邦管理法规.FDA食品法规(2001版)[M].中国轻工业上海设计院,译.北京:中国轻工业出版社,2003:100-199.
[26]H?NSCH R,LANG C,RENNENBERG H,et al.Significance of plant sulfite oxidase[J].Plant Biology,2007,9(5):589-595.DOI:10.1055/s-2007-965433.
[27]KAUFHOLDT D,BAILLIE C K,WILLE T,et al.Prospective posttranslational regulation of plant sulfite oxidase[M]//DE KOK L J,HAWKESFORD M J,RENNENBERG H,et al.Molecular physiology and ecophysiology of sulfur.Switzerland:Springer International Publishing,2015:179-187.DOI:10.1007/978-3-319-20137-5_18.
[28]汪伟,张伟,朱丽琴,等.植物硫化氢生理效应及机制研究进展[J].中国农学通报,2013,29(31):78-82.
[29]ZHANG H,HU L Y,HU K D,et al.Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress[J].Journal of Integrative Plant Biology,2008,50(12):1518-1529.DOI:10.1111/j.1744-7909.2008.00769.x.
[30]GARCíA-MATA C,LAMATTINA L.Hydrogen sulphide,a novel gasotransmitter involved in guard cell signalling[J].New Phytologist,2010,188(4):977-984.DOI:10.1111/j.1469-8137.2010.03465.x.
[31]SCUFFI D,áLVAREZ C,LASPINA N,et al.Hydrogen sulfide generated by L-cysteine desulfhydrase acts upstream of nitric oxide to modulate abscisic acid-dependent stomatal closure[J].Plant Physiology,2014,166(4):2065-2076.DOI:10.1104/pp.114.245373.
[32]HOU Z H,WANG L X,LIU J,et al.Hydrogen sulfide regulates ethylene-induced stomatal closure in Arabidopsis thaliana[J].Journal of Integrative Plant Biology,2013,55(3):277-289.DOI:10.1111/jipb.12004.
[33]ZHANG H,HU S L,ZHANG Z J,et al.Hydrogen sulfide acts as a regulator of flower senescence in plants[J].Postharvest Biology and Technology,2011,60:251-257.DOI:10.1016/j.postharvbio.2011.01.006.
[34]HU L Y,HU S L,WU J,et al.Hydrogen sulfide prolongs postharvest shelf life of strawberry and plays an antioxidative role in fruits[J].Journal of Agriculture and Food Chemistry,2012,60(35):8684-8693.DOI:10.1021/jf300728h.