漆酶LAC表达与鸭梨果心褐变的关系

doi:10.3864/j.issn.0578-1752.2020.24.010

中国农业科学 ›› 2020, Vol. 53 ›› Issue (24): 5073-5080.doi: 10.3864/j.issn.0578-1752.2020.24.010

漆酶LAC表达与鸭梨果心褐变的关系

王梓宇¹(),张引引²,李月圆³,李玲^1,⁴,尤玲玲^1,⁴,李晓雁^1,⁴,晋朝⁵,闫师杰^1,⁴()

¹天津农学院食品科学与生物工程学院,天津 300384
²天津农学院园艺园林学院,天津 300384
³山西农业大学食品科学与工程学院,山西晋中 030801
⁴天津市农副产品深加工技术工程中心,天津 300384
⁵天津森罗科技股份有限公司,天津 300409

收稿日期:2020-02-29 接受日期:2020-10-30 出版日期:2020-12-16 发布日期:2020-12-28
通讯作者: 闫师杰
作者简介:王梓宇,E-mail： prince1893@yeah.net。
基金资助:
国家自然科学基金面上项目(32072278);农业部公益性行业(农业科研专项201303075);天津市林果现代产业技术体系创新团队(ITTFPRS2018009)

Relationship Between LAC Gene Expression and Core Browning of Yali Pear

WANG ZiYu¹(),ZHANG YinYin²,LI YueYuan³,LI Ling^1,⁴,YOU LingLing^1,⁴,LI XiaoYan^1,⁴,JIN Zhao⁵,YAN ShiJie^1,⁴()

¹College of Food Science and Biological Engineering, Tianjin Agricultural University, Tianjin 300384
²College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300384
³College of Food Science and Engineering, Shanxi Agricultural University, Jinzhong 030801, Shanxi
⁴Tianjin Engineering and Technology Research Center of Agricultural Products Processing,, Tianjin 300384
⁵Tianjin CNRO Science & Technology CO. ,Ltd., Tianjin 300409

Received:2020-02-29 Accepted:2020-10-30 Online:2020-12-16 Published:2020-12-28
Contact: ShiJie YAN

摘要/Abstract

摘要：

【目的】探究在不同成熟度和降温贮藏方式下,LAC表达模式与鸭梨果心褐变的关系,为进一步解析鸭梨果心褐变机制提供理论依据。【方法】以鸭梨作为材料,通过对不同成熟度（早采、中采和晚采）的鸭梨进行急速降温（急降）和缓慢降温（缓降）处理,观察贮藏期间鸭梨果心褐变情况,测定漆酶（Laccase,LAC）活性及其基因LAC的相对表达量,研究LAC在鸭梨果心褐变过程中的参与作用。【结果】冷藏60 d时,晚采鸭梨出现褐变,晚采缓降处理的鸭梨果心褐变指数为0.32,是同期急速降温处理的2.56倍;在贮藏90 d时,中采缓降处理的褐变指数是0.24,中采急降处理的褐变指数仅为0.01。各个处理组在贮藏期间LAC活性多数表现为先逐渐升高后下降的趋势,晚采的果实在贮藏60 d时出现活性高峰,褐变发生;早采和中采鸭梨LAC活性高峰均在90 d时出现,褐变程度低于晚采鸭梨。在贮藏期间,缓慢降温处理的LAC活性高于急速降温处理。鸭梨LAC14和LAC7表达量呈先上升后下降的趋势,LAC6表现为先下降后上升再降低的变化趋势;中采、晚采鸭梨在贮藏60 d时,LAC14和LAC7表达量最高。【结论】与缓慢降温相比,急速降温处理减少了鸭梨果心褐变的发生。在整个贮藏期间,LAC活性呈先上升后下降然后又上升的趋势,其峰值时的活性高低次序为：晚采>中采>早采,这与果心褐变趋势一致;LAC在鸭梨果心褐变过程中上调表达。相比缓慢降温处理,急速降温处理具有较低的LAC7、LAC14和LAC6表达量。急速降温结合适时采收能够抑制LAC的上调表达,减少鸭梨果心褐变发生。

关键词: 鸭梨, 果心褐变, 降温方式, 漆酶, LAC

Abstract:

【Objective】 The aim of this study was to explore the relationship between LAC gene expression patterns and core browning of Yali pear under different maturity and cooling storage methods, so as to provide a theoretical basis for further analysis of the mechanism of Yali core browning. 【Method】 Using Yali pear as the material, the Yali pears of different maturity levels (early harvesting, middle harvesting and late harvesting) were treated with rapid cooling and slow cooling to observe the core of the pear during storage. For browning situation, the activity of LAC and the relative expression of LAC gene were determined, and the role of LAC gene in the browning process of Yali pear core was studied. 【Result】 When the Yali pear samples were refrigerated for 60 days, the late-harvested pears browned, and the browning index of late-harvested pears gradually decreased was 0.32, which was 2.56 times the rapid temperature drop during the same period; after 90 days of storage, the middle-harvested browning index decreased slowly, and the index was 0.24, and the browning index of the medium mining was only 0.01. During the storage period, the LAC activity of each treatment group increased gradually and then decreased. The late harvested fruits showed a peak activity and browning occurred after 60 days of storage. The peak of LAC activity of early harvesting and middle harvesting Yali pears appeared in 90 days, and the degree of browning was lower than that of late harvesting Yali. During the storage period, the activity of slow cooling LAC was higher than that of rapid cooling. The LAC14 and LAC7 gene expressions of Yali pears increased first and then decreased. LAC6 realized a change trend of first decline, then increased and then decreased; the expression of LAC14 and LAC7 genes reached the highest when the middle and late harvested pears were stored for 60 days. 【Conclusion】 Compared with slow cooling, rapid cooling treatment reduced the browning of Yali fruit core. During the whole storage period, the activity of LAC showed a trend of rising first, then falling, and then rising again, and the order of activity at its peak was: late harvesting>medium harvesting>early harvesting, which was consistent with the browning trend of the core. LAC was in the Yali fruit Up-regulated expression during heart browning. The rapid cooling was slower than the slow cooling, and the expression of LAC7, LAC14 and LAC6 genes was lower. A timely harvesting with the rapid cooling could inhibit the up-regulated expression of LAC and reduce the browning of Yali pear core.

Key words: Yali pear, core browning, cooling method, laccase, LAC gene

王梓宇,张引引,李月圆,李玲,尤玲玲,李晓雁,晋朝,闫师杰. 漆酶LAC表达与鸭梨果心褐变的关系[J]. 中国农业科学, 2020, 53(24): 5073-5080.

WANG ZiYu,ZHANG YinYin,LI YueYuan,LI Ling,YOU LingLing,LI XiaoYan,JIN Zhao,YAN ShiJie. Relationship Between LAC Gene Expression and Core Browning of Yali Pear[J]. Scientia Agricultura Sinica, 2020, 53(24): 5073-5080.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2020.24.010

https://www.chinaagrisci.com/CN/Y2020/V53/I24/5073

图/表 4

表1

图1

图2

图3

参考文献 32

[1]	韩艳文, 廉双秋, 韩云云, 池明, 闫师杰. 鸭梨POD基因的原核表达及成熟度和降温方法对其表达量的影响. 食品科学, 2017,38(9):40-45. doi: 10.1111/jfds.1973.38.issue-1
	HAN Y W, LIAN S Q, HAN Y Y, CHI M, YAN S J. Prokaryotic expression of POD gene from ‘Yali’ pears and effects of maturity and cooling rate on the expression of POD gene during storage. Food Science, 2017,38(9):40-45. (in Chinese) doi: 10.1111/jfds.1973.38.issue-1
[2]	闫洪波, 程玉豆, 何近刚, 葛文雅, 杨坤, 关军锋. 鸭梨PAL克隆及其在果实发育和机械伤害过程中的表达. 中国农业科学, 2014,47(21):4341-4348. doi: 10.3864/j.issn.0578-1752.2014.21.019
	YAN H B, CHENG Y D, HE J G, GE W Y, YANG K, GUAN J F. Cloning of PAL gene in ‘Yali’ pear and its expression during fruit development and wounding. Scientia Agricultura Sinica, 2014,47(21):4341-4348. (in Chinese) doi: 10.3864/j.issn.0578-1752.2014.21.019
[3]	闫师杰, 陈计峦, 梁丽雅, 汪政富, 胡小松. 降温方法对不同采收成熟度鸭梨某些生理指标的影响. 中国食品学报, 2008,8(4):96-101.
	YAN S J, CHEN J L, LIANG L Y, WANG Z F, HU X S. Effects of different cooling methods on some physiological indexes of different maturity Yali pear after harvest. Journal of Chinese Institute of Food Science and Technology, 2008,8(4):96-101. (in Chinese)
[4]	吴耕西, 周宏伟, 汪建民. 鸭梨酶促褐变的生化机制及底物鉴定. 园艺学报, 1992,19(3):198-202 .
	WU G X, ZHOU H W, WANG J M. Biochemical mechanism and substrate identification of enzymatic browning of ‘Yali’ pears. Acta Horticulturae Sinica, 1992,19(3):198-202. (in Chinese)
[5]	闫师杰, 何爱红, 胡小松. 鸭梨果实采后逆境伤害发生机理的研究进展. 保鲜与加工, 2005,5(4):3-6.
	YAN S J, HE A H, HU X S. Advances on mechanism of stress injury of postharvest Chinese Yali pear fruit. Storage and Process, 2005,5(4):3-6. (in Chinese)
[6]	霍君生, 佟代言, 刘彩莉, 史吉平. 鸭梨果心褐变过程中膜脂过氧化及细胞内膜粘度的变化. 园艺学报, 1995,22(3):221-224.
	HUO J S, TONG D Y, LIU C L, SHI J P. Changes of superoxidation and endomembrane microviscosity of the core of Ya pears during browning development. Acta Horticulturae Sinica, 1995,22(3):21-224. (in Chinese)
[7]	王静. 细胞区室化功能破坏导致的果蔬酶促褐变机理及控制方法. 陕西农业科学, 2014,60(8):62-65.
	WANG J. Enzymatic browning mechanism and control method of fruit and vegetable caused by cell compartmentalization. Shaanxi Journal of Agricultural Sciences, 2014,60(8):62-65. (in Chinese)
[8]	周向军, 杨金龙, 路宛如. 莴笋多酚氧化酶、过氧化物酶的特性及抑制作用研究. 食品工业科技, 2015,36(5):166-174.
	ZHOU X J, YANG J L, LU W R. Properties and inhibitions of polyphenoloxidase and peroxidase from Asparagusplettuce. Science and Technology of Food Industry, 2015,36(5):166-174. (in Chinese)
[9]	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.
	HE L H. Postharvest pear polyPhenoloxidase (PPO) is ozymes and genetic relationship with the browning of fruit[D]. Shihezi: Shihezi University, 2010. (in Chinese)
[10]	李晓丹, 宋方圆, 朱文嫱, 梁丽雅, 闫师杰. 鸭梨采后果肉LOX同工酶变化及其与果实褐变关系的研究. 华北农学报, 2013,28(4):214-217. doi: 10.3969/j.issn.1000-7091.2013.04.039
	LI X D, SONG F Y, ZHU W Q, LIANG L Y, YAN S J. Study on LOX isozymes changes of Yali pear pulp after harvest and its relationship with fruit browning. Acta Agriculturae Boreali-Sinica, 2013,28(4):214-217. (in Chinese) doi: 10.3969/j.issn.1000-7091.2013.04.039
[11]	樊晓岚, 李月圆, 张引引, 韩云云, 李玲, 闫师杰, 肖丽霞. 降温方式对鸭梨采后FAD及LOX基因表达及其褐变的影响. 食品科学, 2019,40(5):222-227.
	FAN X L, LI Y Y, ZHANG Y Y, HAN Y Y, LI L, YAN S J, XIAO L X. Effect of cooling method on FAD and LOX gene expression and core browning in postharvest Yali pears. Food Science, 2019,40(5):222-227. (in Chinese)
[12]	MANGARAJ S, GOSWAMI T K, PANDA D K. Modeling of gas transmission properties films used for MA packaging of fruits. Journal of Food Science and Technology-Mysore, 2015,52(9):5456-5469. doi: 10.1007/s13197-014-1682-2
[13]	OPARA U L, CALEB O J, UCHECHUKWU-AGUA A D. Evaluating the impacts of selected packaging materials on the quality attributes of cassava flour. Journal of Food Science, 2016,81(2):324-331.
[14]	万云洋, 杜予民. 漆酶结构与催化机理. 化学通报, 2007,70(9):662-670.
	WAN Y Y, DU Y M. Structure and catalytic mechanism of Laccases. Chemistry, 2007,70(9):662-670. (in Chinese)
[15]	FANG F, ZHANG X L, LUO H H, ZHOU J J, GONG Y H, LI W J, SHI Z W, HE Q, WU Q, LI L, JIANG L L, CAI Z G, OREN-SHAMIR M, ZHANG Z Q, PANG X Q. An intracellular Laccase is responsible for the epicatechin mediated anthocyanin degradation of litchifruit pericarp. Plant Physiology, 2015,169(4):2391-2408. doi: 10.1104/pp.15.00359 pmid: 26514808
[16]	蒋跃明. 荔枝果实采后果皮褐变的研究[D]. 广州: 中山大学, 1999.
	JIANG Y M. Study on the browning of litchi fruits [D]. Guangzhou: Sun Yat-sen University, 1999. (in Chinese)
[17]	TURLAPATI P V, KIM K W, DAVIN L B, LEWIS N G. The laccase multigene family in Arabidopsis thaliana: Towards addressing the mystery of their gene function (s). Planta, 2011,233:439-470. doi: 10.1007/s00425-010-1298-3
[18]	LE ROY J, BLERVACQ AS, CREACH A, HUSS B, HAWKINS S, NEUTELINGS G. Spatial regulation of monolignol biosynthesis and Laccase genes control developmental and stress-related lignin in flax. BMC Plant Biology, 2017,17:124. doi: 10.1186/s12870-017-1072-9 pmid: 28705193
[19]	WANG J H, FENG J J, JIA W T, FAN P X, BAO H X, LI S Z, LI Y X. Genome-wide identification of sorghum bicolor laccases reveals potential targets for lignin modification. Frontiers in Plant Science, 2017,8:714. doi: 10.3389/fpls.2017.00714 pmid: 28529519
[20]	LIU Q Q, ZHENG L, HE F, ZHAO F J, SHEN Z G. Transcriptional and physiological analyses identify a regulatory role for hydrogen peroxide in the lignin biosynthesis of copper-stressed rice roots. Plant and Soil, 2014,387:323-336. doi: 10.1007/s11104-014-2290-7
[21]	BRYAN A C, JAWDY S, GUNTER L, GJERSING E, SYKES R, HINCHEE M A W, WINKELER K A, COLLINS C M, ENGLE N, TSCHAPLINSKI T J. Knockdown of a Laccase in Populus deltoides confers altered cell wall chemistry and increased sugar release. Plant Biotechnology Journal, 2016,14:2010-2020. doi: 10.1111/pbi.12560 pmid: 26997157
[22]	YAN S J, LI L, HE L H, LIANG L Y, LI X D. Maturity and cooling rate affects browning, polyphenol oxidase activity and gene expression of ‘Yali’ pears during storage. Postharvest Biology and Technology, 2013,85:39-44. doi: 10.1016/j.postharvbio.2013.04.016
[23]	FLOCH C, ALARCON-GUTIERREZ E, CRIQUET S. ABTS assay of phenol oxidase activity in soil. Journal of Microbiological Methods, 2007,71(3):319-324. doi: 10.1016/j.mimet.2007.09.020 pmid: 18006094
[24]	闫洪波, 葛文雅, 杨坤, 程玉豆, 关军锋. 鸭梨肌动蛋白2基因克隆和表达分析. 河北农业大学学报, 2012,35(1):27-30.
	YAN H B, GE W Y, YANG K, CHENG Y D, GUAN J F. Cloning and expression analysis of Actin2 gene in Yali pear. Journal of Agricultural University of Hebei, 2012,35(1):27-30. (in Chinese)
[25]	贺立红, 宾金华. 高等植物中的多酚氧化酶. 植物生理学通讯, 2001,37(4):340-345.
	HE L H, BIN J H. Polyphenol oxidase in higher plants. Plant Physiology Communications, 2001,37(4):340-345. (in Chinese)
[26]	张琥, 王金勇. 红外与漂烫处理下马铃薯多酚氧化酶的失活机理研究. 食品工业, 2017,38(2):9-13.
	ZHANG H, WANG J Y. Study on inactivation mechanism of potato PPO under infrared processing and blanching processing. The Food Industry, 2017,38(2):9-13. (in Chinese)
[27]	NIU B, GUO L, ZHAO M M, LUO T, ZHANG R, ZHANG F L, HOU P, ZHANG Y, XU Y, WANG S H. Molecular cloning, characterization, and expression of an ω-3 fatty acid desaturase gene from Sapium sebiferum. Journal of Bioscience and Bioengineering, 2008,106(4):375-380. doi: 10.1263/jbb.106.375 pmid: 19000614
[28]	DING C K, CHACHIN K, UEDA Y, IMAHORI Y. Purification and properties of polyphenol oxidase from loquat fruit. Journal of Agricutural and Food Chemistry, 1998,46:4144-4149. doi: 10.1021/jf980093s
[29]	林河通, 席玙芳, 陈绍军. 果实贮藏期间的酶促褐变. 福州大学学报(自然科学版), 2002,30(Z1):696-703.
	LIN H T, XI Y F, CHEN S J. A review of enzymatic browning in fruit during storage. Journal of Fuzhou University (Natural Science Edtion), 2002,30(Z1):696-703. (in Chinese)
[30]	刘保华, 肖茜, 冯超, 孙进华, 王家保. 荔枝漆酶基因LcLAC的克隆与表达分析. 园艺学报, 2012,39(5):853-860.
	LIU B H, XIAO Q, FENG C, SUN J H, WANG J B. Cloning and expression analysis of the Laccase gene from Litchi chinensis. Acta Horticulturae Sinica, 2012,39(5):853-860. (in Chinese)
[31]	帅良, 赵昱清, 廖玲燕, 宋幕波, 朱东建, 蔡文, 段振华, 吴振先, 韩冬梅. 龙眼漆酶基因(Dllac)的克隆及表达分析. 食品工业科技, 2017,38(13):95-100.
	SHUAI L, ZHAO Y Q, LIAO L Y, SONG M B, ZHU D J, CAI W, DUAN Z H, WU Z X, HAN D M. Cloning and expression analysis of the Laccase gene (Dllac) from Dimocarpus longan. Science and Technology of Food Industry, 2017,38(13):95-100. (in Chinese)
[32]	周一鹏. 柑橘漆酶基因家族的鉴定及抗逆性研究[D]. 扬州: 扬州大学, 2018.
	ZHOU Y P. Genome-wide identification and characterization of Laccase gene family and resistance study in Citrus sinensis[D]. Yangzhou: Yangzhou University, 2018. (in Chinese)

基因名称 Gene name	基因号 Gene ID	引物序列（5'-3'） Primer sequence (5'-3')	扩增片段大小 Fragment size (bp)
pbACT	NW_008988356.1	F：TGGTATAGTGAGCAACTGGGATGA R：CCTCTGTGAGCAGAACTGGGTG	104
pbLAC6	NW_008988414.1	F：AACACCAAGGACATCGTGACCATC R：TGACGCCGTGCCAGTGAATTG	184
pbLAC14	NW_008988132.1	F：CGCAGTTCACACCGAGATCATCC R：TTGCTAGGCTTCTGAAGTGCTTGG	85
pbLAC7	NW_008988996.1	F：TTGTTGGGTATGGGAGTG R：AGGAGGATGAGGAAGGGT	242

漆酶LAC表达与鸭梨果心褐变的关系

Relationship Between LAC Gene Expression and Core Browning of Yali Pear

RichHTML

PDF

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 32

相关文章 9

Metrics

本文评价

推荐阅读 0

[1]	岳盈肖,何近刚,赵江丽,闫子茹,程玉豆,武肖琦,王永霞,关军锋. 窖藏和冷藏条件下鸭梨挥发性物质及其相关基因表达分析[J]. 中国农业科学, 2021, 54(21): 4635-4649.
[2]	杜艳民,王文辉,贾晓辉,佟伟,王阳,张鑫楠. 不同O₂浓度对鸭梨采后生理代谢及贮藏品质的影响[J]. 中国农业科学, 2020, 53(23): 4918-4928.
[3]	曹可可，刘宁，马双新，曹志艳，梁东旭，柴江婷，董金皋. 大斑刚毛座腔菌高产漆酶条件的响应面优化及酶学特性[J]. 中国农业科学, 2015, 48(11): 2165-2175.
[4]	李朋朋，刘宁，张玉星，彭建营，董金皋. 鸭梨几丁质PbChiIV的克隆与表达[J]. 中国农业科学, 2015, 48(11): 2279-2286.
[5]	陈鹏, 屈明博, 杨君, 杨青. 亚洲玉米螟两种漆酶基因OfLac1和OfLac2 cDNA 的克隆及基因表达[J]. 中国农业科学, 2014, 47(7): 1341-1350.
[6]	闫洪波，程玉豆，何近刚，葛文雅，杨坤，关军锋. 鸭梨PAL克隆及其在果实发育和机械伤害过程中的表达[J]. 中国农业科学, 2014, 47(21): 4341-4348.
[7]	詹旭,曹志艳,邢继红,董金皋 . 植物病原真菌产漆酶菌株的筛选[J]. 中国农业科学, 2011, 44(4): 723-729 .
[8]	鞠志国,原永兵,刘成连,战淑敏,辛士海. 急降温对活性氧和梨果心褐变的影响[J]. 中国农业科学, 1994, 27(05): 77-81 .
[9]	陈昆松,于樑,周山涛. 鸭梨果实气调贮藏过程CO#-2伤害机理初探[J]. 中国农业科学, 1991, 24(05): 83-88 .