降温方式对鸭梨采后FAD及LOX基因表达及其褐变的影响
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摘要
为了研究急速降温处理与缓慢降温处理时,FAD与LOX基因表达量对鸭梨果心褐变的影响,以适时(9月中旬)采收鸭梨为材料,对鸭梨贮藏中果心部位脂肪酸含量、ω-3脂肪酸去饱和酶(ω-3 fatty acid desaturase,ω-3FAD)基因表达量、脂氧合酶(lipoxygenase,LOX)活力及基因表达量进行测定。结果表明:贮藏期间,急速降温处理组果心褐变指数明显高于缓慢降温处理组,并且急速降温处理组受到低温胁迫,机体产生抗冷机制,导致果心不饱和脂肪酸(尤其是亚麻酸)含量,ω-3FAD1、FAD2表达量与LOX活力均有不同程度的增加,并且低温激活LOX基因表达,加速膜脂过氧化,降低鸭梨品质。因此,对于适时采收的鸭梨,应采取缓慢降温的方式进行贮藏,以提高鸭梨贮藏品质。
In order to explore the effects of rapid and slow cooling on ω-3 fatty acid desaturase(ω-3 FAD) and lipoxygenase(LOX) gene expression and core browning in Yali pears, Hebei grown Yali pears(Pyrus bretschneideri Rehd cv. Yali) were chosen to determine the contents of fatty acids and the gene expression and activities of FAD and LOX. The results showed that the core browning index of the fruits subjected to rapid cooling was signi?cantly higher than that of those subjected to slow cooling during storage. Cold resistance occurred in the rapid cooling group, resulting in increased contents of unsaturated fatty acids, up-regulated expression levels of ω-3 FAD1 and FAD2, and enhanced activities of LOX. Moreover,low-temperature stress activated the expression of LOX gene, accelerated membrane lipid peroxidation, and reduced fruit quality. Therefore, timely harvested Yali pears should be cooled slowly in order to improve the storage quality.
In order to explore the effects of rapid and slow cooling on ω-3 fatty acid desaturase(ω-3 FAD) and lipoxygenase(LOX) gene expression and core browning in Yali pears, Hebei grown Yali pears(Pyrus bretschneideri Rehd cv. Yali) were chosen to determine the contents of fatty acids and the gene expression and activities of FAD and LOX. The results showed that the core browning index of the fruits subjected to rapid cooling was signi?cantly higher than that of those subjected to slow cooling during storage. Cold resistance occurred in the rapid cooling group, resulting in increased contents of unsaturated fatty acids, up-regulated expression levels of ω-3 FAD1 and FAD2, and enhanced activities of LOX. Moreover,low-temperature stress activated the expression of LOX gene, accelerated membrane lipid peroxidation, and reduced fruit quality. Therefore, timely harvested Yali pears should be cooled slowly in order to improve the storage quality.
引文
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[8]叶静,胡海涛,王长春,等.植物脂肪酸去饱和酶与抗冷性研究进展[J].广东农业科学,2011,38(12):131-134.DOI:10.3969/j.issn.1004-874X.2011.12.048.
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[13]杨青珍.猕猴桃果实采后冷害发生生理机制及调控作用[D].杨凌:西北农林科技大学,2013:35-39.
[14]丁一,王华森,于超.番茄内质网ω-3脂肪酸去饱和酶基因原核表达载体的构建与鉴定[J].生物技术通报,2012(1):79-82.DOI:10.13560/j.cnki.biotech.bull.1985.2012.01.013.
[15]魏雯雯.桃果实冷害相关FAD基因家族的表达调控[D].杭州:浙江大学,2011:24-30.
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[17]张姣姣,郝晓磊,李喜宏,等.热空气协同茉莉酸甲酯处理对冷藏石榴冷害及果实品质的影响[J].中国果树,2016(5):29-33.DOI:10.16626/j.cnki.issn1000-8047.2016.05.005.
[18]何利华,江英,闫师杰,等.降温方法对不同采收成熟度鸭梨果心PPO活性及褐变影响的研究[J].保鲜与加工,2010,10(3):42-45.
[19]何子顺,李芳芳,张绍铃,等.套袋对‘库尔勒香梨’果实中游离脂肪酸和游离氨基酸含量的影响[J].果树学报,2016,23(7):804-813.DOI:10.13925/j.cnki.gsxb.20150510.
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[21]GASIC K,HERNANDEZ A,KORBAN S S.RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction[J].Plant Molecular Biology Reporter,2004,22(4):437-438.DOI:10.1007/BF02772687.
[22]李晓丹,宋方圆,朱文嫱,等.鸭梨采后果肉LOX同工酶变化及其与果实褐变关系的研究[J].华北农学报,2013,28(4):214-217.
[23]高慧,饶景萍.冷害对贮藏油桃膜脂脂肪酸及相关酶活性的影响[J].西北植物学报,2007(4):4710-4714.
[24]秦晓惠,李凯文,段志坤.植物冷胁迫信号研究进展[J].分子植物育种,2018,16(21):7187-7194.DOI:10.13271/j.mpb.016.007187.
[25]SAPITNITSKAYA M.Postharvest heat and conditioning treatments activate different molecular responses and reduce chilling injuries in grapefruit[J].Journal of Experimental Botany,2006,57(12):2943-2953.DOI:10.1093/jxb/erl055.
[26]孟坤.柿脂氧合酶及其相关基因与果实成熟软化的关系研究[D].杨凌:西北农林科技大学,2016:6-11.
[27]韩云云,宋方圆,韩艳文,等.不同采收贮藏条件下鸭梨果实LOX基因表达及其与果心褐变的关系[J].食品科学,2016,37(18):216-222.DOI:10.7506/spkx1002-6630-201618035.
[28]何利华.鸭梨采后多酚氧化酶(PPO)同工酶及基因与果实褐变关系的研究[D].石河子:石河子大学,2010:2-10.
[29]张爱琳,胡小松,李晓丹,等.降温方法对鸭梨采后果肉与果心抗冷性的影响[J].食品与机械,2011,27(2):114-118.DOI:10.3969/j.issn.1003-5788.2011.02.035.
[30]ZHANG C F,TIAN S P.Crucial contribution of membrane lipids’ unsaturation to acquisition of chilling-tolerance in peach fruit stored at 0℃[J].Food Chemistry,2008,115(2):405-411.DOI:10.1016/j.foodchem.2008.12.021.
[31]KODAMA H,HORIGUCHI G,NISHIUCHI T,et al.Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low-temperature tolerance to young tobacco leaves[J].Plant Physiology,1995,107(4):1177-1185.DOI:10.1104/pp.107.4.1177.
[32]SIDDIQUI K S,CAVICCHIOLI R.Cold-adapted enzymes[J].Annual Review of Biochemistry,2006,75(1):403-433.DOI:10.1146/annurev.biochem.75.103004.142723.
[33]DE VIRVILLE J D,CANTREL C,BOUSQUET A L,et al.Homeoviscous and functional adaptations of mitochondrial membranes to growth temperature in soybean seedlings[J].Plant Cell&Environment,2002,25(10):1289-1297.DOI:10.1046/j.1365-3040.2002.00901.x.
[34]MATOS A R,HOURTON-CABASSA C,CI?EK D,et al.Alternative oxidase involvement in cold stress response of Arabidopsis thaliana fad2 and FAD3+cell suspensions altered in membrane lipid composition[J].Plant and Cell Physiology,2007,48(6):856-865.DOI:10.1093/pcp/pcm061.
[35]梁芸志,陈存坤,吴昊,等.不同预冷温度对采后番茄贮藏品质的影响[J].食品研究与开发,2018,39(13):188-193;200.
[36]刘双禄.低温处理对河套蜜瓜贮藏品质的影响[J].河套学院论坛,2018,15(2):87-91.
[37]张波,李鲜,陈昆松.脂氧合酶基因家族成员与果实成熟衰老研究进展[J].园艺学报,2007(1):245-250.DOI:10.16420/j.issn.0513-353x.2007.01.054.
[38]何婷,赵谋明,崔春.脂肪氧合酶的酶学特性及其活性抑制机理的研究进展[J].食品工业科技,2008(2):291-293;298.DOI:10.13386/j.issn1002-0306.2008.02.070.
[39]LIU H,SONG L L,YOU Y L,et al.Cold storage duration affects litchi fruit quality,membrane permeability,enzyme activities and energy charge during shelf time at ambient temperature[J].Postharvest Biology&Technology,2011,60(1):24-30.DOI:10.1016/j.postharvbio.2010.11.008.
[40]PONGPRASERT N,SEKOZAWA Y,SUGAYA S,et al.A novel postharvest UV-C treatment to reduce chilling injury(membrane damage,browning and chlorophyll degradation)in banana peel[J].Scientia Horticulturae,2011,130(1):73-77.DOI:10.1016/j.scienta.2011.06.006.
[41]LIN Y F,LIN H T,ZHANG S,et al.The role of active oxygen metabolism in hydrogen peroxide-induced pericarp browning of harvested longan fruit[J].Postharvest Biology and Technology,2014,96:42-48.DOI:10.1016/j.postharvbio.2014.05.001.
[42]KOU X H,WU M S,LI L,et al.Effects of CaCl2 dipping and pullulan coating on the development of brown spot on‘Huangguan’pears during cold storage[J].Postharvest Biology and Technology,2015,99:63-72.DOI:10.1016/j.postharvbio.2014.08.001.
[2]闫师杰,陈计峦,梁丽雅,等.降温方法对不同采收成熟度鸭梨某些生理指标的影响[J].中国食品学报,2008,8(4):96-101.DOI:10.16429/j.1009-7848.2008.04.020.
[3]卢铁道,王刚柱.鸭梨贮藏技术[J].现代农业科技,2015(8):296;298.
[4]梁丽雅,胡小松,何爱红,等.降温方法对不同成熟度鸭梨果肉脂氧合酶活性和膜脂脂肪酸的影响[J].食品科技,2012,37(4):35-39.DOI:10.13684/j.cnki.spkj.2012.04.005.
[5]王君.鸭梨、黄金梨采后褐变生理及抗褐变研究[D].保定:河北农业大学,2006:12-17.DOI:10.7666/d.y933480.
[6]SAKAMOTO T,MURATA N.Regulation of the desaturation of fatty acids and its role in tolerance to cold and salt stress[J].Current Opinion in Microbiology,2002,5(2):206-210.DOI:10.1016/S1369-5274(02)00306-5.
[7]WANG L J,LI S H.Salicylic acid-induced heat or cold tolerance in relation to Ca2+homeostasis and antioxidant systems in young grape plants[J].Plant Science,2006,170(4):685-694.DOI:10.1016/j.plantsci.2005.09.005.
[8]叶静,胡海涛,王长春,等.植物脂肪酸去饱和酶与抗冷性研究进展[J].广东农业科学,2011,38(12):131-134.DOI:10.3969/j.issn.1004-874X.2011.12.048.
[9]BISWAS P,EAST A R,HEWETT E,et al.Interpreting textural changes in low temperature stored tomatoes[J].Postharvest Biology and Technology,2014,87:140-143.DOI:10.1016/j.postharvbio.2013.08.018.
[10]高岩,郭东林,郭长虹.三烯脂肪酸在高等植物逆境胁迫应答中的作用[J].分子植物育种,2010,8(2):365-369.
[11]KANAMOTO H,TAKEMURA M,OHYAMA K.Cloning and expression of three lipoxygenase genes from liverwort,Marchantia polymorpha L.,in Escherichia coli[J].Phytochemistry,2012,77:97-102.DOI:10.1016/j.phytochem.2012.02.009.
[12]霍君生,佟代言,刘彩莉,等.鸭梨果心褐变过程中膜质过氧化及细胞内膜微粘度的变化[J].园艺学报,1995(3):221-224.
[13]杨青珍.猕猴桃果实采后冷害发生生理机制及调控作用[D].杨凌:西北农林科技大学,2013:35-39.
[14]丁一,王华森,于超.番茄内质网ω-3脂肪酸去饱和酶基因原核表达载体的构建与鉴定[J].生物技术通报,2012(1):79-82.DOI:10.13560/j.cnki.biotech.bull.1985.2012.01.013.
[15]魏雯雯.桃果实冷害相关FAD基因家族的表达调控[D].杭州:浙江大学,2011:24-30.
[16]赖志宸,林玉玲.野生蕉ω-3脂肪酸去饱和酶基因FAD7密码子偏性分析[J].福建农业学报,2017,32(5):503-507.DOI:10.19303/j.issn.1008-0384.2017.05.008.
[17]张姣姣,郝晓磊,李喜宏,等.热空气协同茉莉酸甲酯处理对冷藏石榴冷害及果实品质的影响[J].中国果树,2016(5):29-33.DOI:10.16626/j.cnki.issn1000-8047.2016.05.005.
[18]何利华,江英,闫师杰,等.降温方法对不同采收成熟度鸭梨果心PPO活性及褐变影响的研究[J].保鲜与加工,2010,10(3):42-45.
[19]何子顺,李芳芳,张绍铃,等.套袋对‘库尔勒香梨’果实中游离脂肪酸和游离氨基酸含量的影响[J].果树学报,2016,23(7):804-813.DOI:10.13925/j.cnki.gsxb.20150510.
[20]SHEN Wenyun,LI J Q,DAUK M,et al.Metabolic and transcriptional responses of glycerolipid pathways to aperturbation of glycerol3-phosphate metabolism in Arabidopsis[J].Journal of Biological Chemistry,2010,285(30):22957-22965.DOI:10.1074/jbc.M109.097758.
[21]GASIC K,HERNANDEZ A,KORBAN S S.RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction[J].Plant Molecular Biology Reporter,2004,22(4):437-438.DOI:10.1007/BF02772687.
[22]李晓丹,宋方圆,朱文嫱,等.鸭梨采后果肉LOX同工酶变化及其与果实褐变关系的研究[J].华北农学报,2013,28(4):214-217.
[23]高慧,饶景萍.冷害对贮藏油桃膜脂脂肪酸及相关酶活性的影响[J].西北植物学报,2007(4):4710-4714.
[24]秦晓惠,李凯文,段志坤.植物冷胁迫信号研究进展[J].分子植物育种,2018,16(21):7187-7194.DOI:10.13271/j.mpb.016.007187.
[25]SAPITNITSKAYA M.Postharvest heat and conditioning treatments activate different molecular responses and reduce chilling injuries in grapefruit[J].Journal of Experimental Botany,2006,57(12):2943-2953.DOI:10.1093/jxb/erl055.
[26]孟坤.柿脂氧合酶及其相关基因与果实成熟软化的关系研究[D].杨凌:西北农林科技大学,2016:6-11.
[27]韩云云,宋方圆,韩艳文,等.不同采收贮藏条件下鸭梨果实LOX基因表达及其与果心褐变的关系[J].食品科学,2016,37(18):216-222.DOI:10.7506/spkx1002-6630-201618035.
[28]何利华.鸭梨采后多酚氧化酶(PPO)同工酶及基因与果实褐变关系的研究[D].石河子:石河子大学,2010:2-10.
[29]张爱琳,胡小松,李晓丹,等.降温方法对鸭梨采后果肉与果心抗冷性的影响[J].食品与机械,2011,27(2):114-118.DOI:10.3969/j.issn.1003-5788.2011.02.035.
[30]ZHANG C F,TIAN S P.Crucial contribution of membrane lipids’ unsaturation to acquisition of chilling-tolerance in peach fruit stored at 0℃[J].Food Chemistry,2008,115(2):405-411.DOI:10.1016/j.foodchem.2008.12.021.
[31]KODAMA H,HORIGUCHI G,NISHIUCHI T,et al.Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low-temperature tolerance to young tobacco leaves[J].Plant Physiology,1995,107(4):1177-1185.DOI:10.1104/pp.107.4.1177.
[32]SIDDIQUI K S,CAVICCHIOLI R.Cold-adapted enzymes[J].Annual Review of Biochemistry,2006,75(1):403-433.DOI:10.1146/annurev.biochem.75.103004.142723.
[33]DE VIRVILLE J D,CANTREL C,BOUSQUET A L,et al.Homeoviscous and functional adaptations of mitochondrial membranes to growth temperature in soybean seedlings[J].Plant Cell&Environment,2002,25(10):1289-1297.DOI:10.1046/j.1365-3040.2002.00901.x.
[34]MATOS A R,HOURTON-CABASSA C,CI?EK D,et al.Alternative oxidase involvement in cold stress response of Arabidopsis thaliana fad2 and FAD3+cell suspensions altered in membrane lipid composition[J].Plant and Cell Physiology,2007,48(6):856-865.DOI:10.1093/pcp/pcm061.
[35]梁芸志,陈存坤,吴昊,等.不同预冷温度对采后番茄贮藏品质的影响[J].食品研究与开发,2018,39(13):188-193;200.
[36]刘双禄.低温处理对河套蜜瓜贮藏品质的影响[J].河套学院论坛,2018,15(2):87-91.
[37]张波,李鲜,陈昆松.脂氧合酶基因家族成员与果实成熟衰老研究进展[J].园艺学报,2007(1):245-250.DOI:10.16420/j.issn.0513-353x.2007.01.054.
[38]何婷,赵谋明,崔春.脂肪氧合酶的酶学特性及其活性抑制机理的研究进展[J].食品工业科技,2008(2):291-293;298.DOI:10.13386/j.issn1002-0306.2008.02.070.
[39]LIU H,SONG L L,YOU Y L,et al.Cold storage duration affects litchi fruit quality,membrane permeability,enzyme activities and energy charge during shelf time at ambient temperature[J].Postharvest Biology&Technology,2011,60(1):24-30.DOI:10.1016/j.postharvbio.2010.11.008.
[40]PONGPRASERT N,SEKOZAWA Y,SUGAYA S,et al.A novel postharvest UV-C treatment to reduce chilling injury(membrane damage,browning and chlorophyll degradation)in banana peel[J].Scientia Horticulturae,2011,130(1):73-77.DOI:10.1016/j.scienta.2011.06.006.
[41]LIN Y F,LIN H T,ZHANG S,et al.The role of active oxygen metabolism in hydrogen peroxide-induced pericarp browning of harvested longan fruit[J].Postharvest Biology and Technology,2014,96:42-48.DOI:10.1016/j.postharvbio.2014.05.001.
[42]KOU X H,WU M S,LI L,et al.Effects of CaCl2 dipping and pullulan coating on the development of brown spot on‘Huangguan’pears during cold storage[J].Postharvest Biology and Technology,2015,99:63-72.DOI:10.1016/j.postharvbio.2014.08.001.