Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (21): 4341-4348.doi: 10.3864/j.issn.0578-1752.2014.21.019

• RESEARCH NOTES • Previous Articles    

Cloning of PAL Gene in ‘Yali’ Pear and Its Expression During Fruit Development and Wounding

YAN Hong-bo1,2,3, CHENG Yu-dou1,3, HE Jin-gang1,3, GE Wen-ya1, 3, YANG Kun1,3, GUAN Jun-feng1,3   

  1. 1Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051
    2Bioscience and Bioengineering School, Hebei University of Economy and Business, Shijiazhuang 050061
    3Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051
  • Received:2014-01-14 Revised:2014-07-18 Online:2014-11-01 Published:2014-11-01

RichHTML

1

PDF

630

Abstract: 【Objective】 The objective of this study is to investigate the phenylalanine ammonia-lyase gene (PAL) sequence characters and expression pattern during fruit development and tissue browning in ‘Yali’ pear.【Method】Got full coding domain sequence of PAL by homology-based cloning, the bioinformatic characteristics of the cloned PAL were analyzed using online service, PAL protein phylogenetic tree was constructed by MEGA 5.1 software, analyzed the expression pattern of different development stages of fruit and tissue browning of ‘Yali’ pear by real-time quantitative RT-PCR. 【Result】 Two PAL genes containing full length CDS were obtained and termed PbPAL1 and PbPAL2, respectively. The full length cDNA of PbPAL1 was 2 232 bp, containing 2 160 bp open reading frame, 60 bp 5′ untranslated region and 12 bp 3′ untranslated region (GeneBank Access Number: GU906268.1). The full length cDNA of PbPAL2 was 2 387 bp, containing 2 163 bp open reading frame, 14 bp 5′ untranslated region and 210 bp 3′ untranslated region (GeneBank Access Number: GU906269.1). Phylogenetic analysis indicated that PbPAL1 and PbPAL2 were classified into two different phylogenetic tree branches. PbPAL1 was highly homologous to PAL of Pyrus communis, while PbPAL2 was highly homologous to PAL of Prunus persica. Although the expression levels of the PbPAL1 and PbPAL2 were different in the peel, flesh and core, all of the expression patterns showed a down trend along with fruit development, and were significantly higher in core than those in peel and flesh at early development stage. Wounding treatment could rapidly induce browning of flesh, while slowly induce the browning of peel. In both flesh and peel, the expression levels of PbPAL1 and PbPAL2 were markedly up-regulated by wounding treatment. In peel, the expression level of PbPAL1 appeared a peak at 6 h after wounding treatment, while PbPAL2 mRNA level significantly increased at 48 h after treatment. In flesh, the expression level of PbPAL1 obviously increased at 1 h after wounding treatment, and PbPAL2 began to rise at 12 h after wounding treatment.【Conclusion】PbPAL1 and PbPAL2 belong to different PAL gene types, and their expression showed a decreasing trend along with fruit development, and up-regulated by wounding, thus, they were involved in tissue browning induced by wounding in ‘Yali’ pear.

Key words: ‘Yali&rsquo, pear, phenylalanine ammonia-lyase, browning, wounding, fruit development

[1]    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 substances determination of enzymic browning of ‘Yali’ pear (Pyrus Bretschneideri Rehd.). Acta Horticulture Sinica, 1992, 19(3): 198-202. (in Chinese)
[2]    Vanholme R, Demedts B, Morreel K, Ralph J, Boerjan W. Lignin biosynthesis and structure. Plant Physiology, 2010, 153(3): 895-905.
[3]    Achnine D R. Natural products and plant disease resistance. Nature, 2001, 411, 843-847.
[4]    Holgre R, Georg E. Structural basis for the entrance into the phenylpropanoid metabolism catalyzed by phenylalanine ammonia- lyase. The Plant Cell, 2004, 16(12): 3426-3436.
[5]    Achnine L, Blancaflor E B, Rasmussen S, Dixon R A. Colocalization of L-phenylalanine ammonialyase and cinnamate 4-hydroxylase for metabolic channeling in phenylpropanoid biosynthesis. The Plant Cell, 2004, 16(11): 3098-3109.
[6]    程水源, 陈昆松, 刘卫红, 杜何为. 植物苯丙氨酸解氨酶基因的表达调控与研究展望. 果树学报, 2003, 20(5): 351-357.
Cheng S S, Chen K S, Liu W H, Du H W. Regulation and expression of the PAL in plant and its outlook. Journal of Fruit Science, 2003, 20(5): 351-357. (in Chinese)
[7]    鞠志国, 刘成连, 原永兵, 戴洪义, 阎升平, 徐军. 莱阳茌梨酚类物质合成的调节及其对果实品质的影响. 中国农业科学, 1993, 26(4): 44-48.
Ju Z G, Liu C L, Yuan Y B, Dai H Y, Yan S P, Xu J. Regulation of phenolics synthesis and its effects on fruit quality of Laiyang Chili (pear). Scientia Agricultura Sinica, 1993, 26(4): 44-48. (in Chinese)
[8]    Huang J L, Gu M, Lai Z, Fan B, Shi K, Zhou Y H, Yu J Q, Chen Z. Functional analysis of the Arabidopsis PAL gene family in plant growth, development, and response to environmental stress. Plant Physiology, 2010, 153(4): 1526-1538.
[9]    Reichert A I, He X Z, Dixon R A. Phenylalanine ammonia-lyase (PAL) from tobacco (Nicotiana tabacum): Characterization of the four tobacco PAL genes and active heterotetrameric enzymes. Biochemical Journal, 2009, 424(2): 233-242.
[10]   Chang A, Lim M H, Lee S W, Robb E J, Nazar R N. Tomato phenylalanine ammonia-lyase gene family, highly redundant but strongly underutilized. The Journal of Biological Chemistry, 2008, 283(48): 33591-33601.
[11]   Liang X W, Dron M, Cramer C L, Dixon R A, Lamb C J. Differential regulation of phenylalanine ammonia-lyase genes during plant development and by environmental cues. The Journal of Biological Chemistry, 1989, 264(24): 14486-14492.
[12]   荣瑞芬, 郭堃, 厉重先, 李丽云, 李祖明. 紫外照射诱导采后番茄苯丙氨酸解氨酶的分离纯化. 北方园艺, 2007, 12: 1-4.
Rong R F, Guo K, Li C X, Li L Y, Li Z M. Purification and properties of PAL from harvested tomato induced by UV-C irradiation. Northern Horticulture, 2007, 12: 1-4. (in Chinese)
[13]   马俊彦, 杨汝德, 敖利刚. 植物苯丙氨酸解氨酶的生物学研究进展. 现代食品科技, 2007, 23(7): 71-74.
Ma J Y, Yang R D, Ao L G. Progress in biological research of phenylalanine ammonialyase. Modern Food Science and Technology, 2007, 23(7): 71-74. (in Chinese)
[14]   Kumar A, Ellis B E. The phenylalanine ammonia-lyase gene family in raspberry. Structure, expression, and evolution. Plant Physiology, 2001, 127(1): 230-239.
[15]   李正国, 高雪, 樊晶, 杨迎伍, 李道高, Kanellis A K. 奉节脐橙果实苯丙氨酸解氨酶活性及其基因表达与果皮褐变的关系. 植物生理与分子生物学学报, 2006, 32(3): 381-386.
Li Z G, Gao X, Fan J, Yang Y W, Li D G, Kanellis A K. The Relationship between activity and gene expression of phenylalanine ammonia-lyase and peel pitting in ‘Fengjie’ navel orange fruits. Journal of Plant Physiology and Molecular Biology, 2006, 32(3): 381-386. (in Chinese)
[16]   张士鸿, 金青, 樊洪, 黄蓓, 蔡永萍, 林毅, 张金云, 徐义流. 砀山酥梨果实苯丙氨酸解氨酶基因cDNA克隆及序列分析. 激光生物学报, 2010, 19(1): 97-103.
Zhang S H, Jin Q, Fan H, Huang B, Cai Y P, Lin Y, Zhang J Y, Xu Y L. Cloning and sequencing of a phenylalanine ammonia-lyase cDNA from Pyrus bretschneideri cv. Acta Laser Biology Sinica, 2010, 19(1): 97-103. (in Chinese)
[17]   Gasic K, Hernandez A, Korban S S. RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction. Plant Molecular Biology Reporter, 2004, 22(4): 437-438.
[18]   林智华, 林程忠, 梁红, 周玲艳. 猕猴桃基因组DNA提取的研究. 安徽农业科学, 2009, 37(3): 996-998.
Lin Z H, Lin C Z, Liang H, Zhou L Y. Study on extraction of Actinidia genomic DNA. Journal of Anhui Agricultural Sciences, 2009, 37(3): 996-998. (in Chinese)
[19]   Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology Evolution, 2011, 28(10): 2731-2739.
[20]   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 of actin2 gene in ‘Yali’ pear. Journal of Hebei Agricultural University, 2012, 35(1): 27-30. (in Chinese)
[21]   Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 2001, 25(4): 402-408.
[22]   张振铭, 施泽斌, 张绍铃, 乔勇进, 陶书田. 砀山酥梨不同发育时期套袋对石细胞发育的影响. 园艺学报, 2007, 43(3): 565-568.
Zhang Z M, Shi Z B, Zhang S L, Qiao Y J, Tao S T. Bagging at different developmental stages on sclereid formation in ‘Dangshan su’ pear fruit. Acta Horticulturae Sinica, 2007, 43(3): 565-568. (in Chinese)
[23]   王斌, 张楠, 闫冲冲, 金青, 林毅, 蔡永萍, 张金云. 套袋对砀山酥梨果实石细胞发育及木质素代谢的影响. 园艺学报, 2013, 40(3): 531-539.
Wang B, Zhang N, Yan C C, Jin Q, Lin Y, Cai Y P, Zhang Y J. Bagging for the development of stone cell and metabolism of lignin in Pyrus bretschneideri ‘Dangshan Suli’. Acta Horticulturae Sinica, 2013, 40(3): 531-539. (in Chinese)
[24]   俞雅琼, 董明, 王旭东, 张国庆, 李娜. 机械损伤对砀山酥梨采后生理生化变化的影响. 保鲜与加工, 2011, 11(3): 10-15.
Yu Y Q, Dong M, Wang X D, Zhang G Q, Li N. Effects of mechanical damage on postharvest physiology and biochemistry of ‘Dangshan’ pear. Storage and Process, 2011, 11(3): 10-15. (in Chinese)
[25]   魏敏, 周会玲, 陈小利, 辛付存. 低温贮藏对鲜切富士苹果褐变的影响. 西北林学院学报, 2011, 26(5): 131-134.
Wei M, Zhou H L, Chen X l, Xin F C. Effect s of storage temperature on browning of fresh cut ’Fuji’ apples. Journal of Northwest A & F University, 2011, 26(5): 131-134. (in Chinese)
[26]   Nguyen T B T, Ketsa S, Van Doorn W G. Relationship between browning and the activities of polyphenol oxidase and phenylalanine ammonia lyase in banana peel during low temperature storage. Postharvest Biology and Technology, 2003, 30(2): 187-193.
[27]   López-Galvez G, Saltveit M, Cantwell M. Wound-induced phenylalanine ammonia lyase activity, factors affecting its induction and correlation with the quality of minimally processed lettuces. Postharvest Biology and Technology, 1996, 9(2):223-233.
[28]   Kato M, Hayakawa Y, Hyodo H, Ikoma Y, Yano M. Wound induced ethylene synthesis and expression and formation of 1-aminocyclopropane- 1-carboxylate (ACC) synthase, ACC oxidase, phenylalanine ammonia- lyase, and peroxidase in wounded mesocarp tissue of Cucurbita maxima. Plant Cell Physiology, 2000, 41(4): 440-447.
[29]   Kao Y, Harding S A, Tsai C. Differential expression of two distinct phenylalanine ammonia-lyase genes in condensed tannin-accumulating and lignifying cells of quaking aspen. Plant Physiology, 2002, 130(2): 796-807.
[1] LI ShiJia,LÜ ZiJing,ZHAO Jin. Identification of R2R3-MYB Subfamily in Chinese Jujube and Their Expression Pattern During the Fruit Development [J]. Scientia Agricultura Sinica, 2022, 55(6): 1199-1212.
[2] XIANG MiaoLian, WU Fan, LI ShuCheng, WANG YinBao, XIAO LiuHua, PENG WenWen, CHEN JinYin, CHEN Ming. Effects of Melatonin Treatment on Resistance to Black Spot and Postharvest Storage Quality of Pear Fruit [J]. Scientia Agricultura Sinica, 2022, 55(4): 785-795.
[3] JIA XiaoHui,ZHANG XinNan,LIU BaiLin,MA FengLi,DU YanMin,WANG WenHui. Effects of Low Oxygen/High Carbon Dioxide Controlled Atmosphere Combined with 1-Methylcyclopropene on Quality of Yuluxiang Pear During Cold Storage [J]. Scientia Agricultura Sinica, 2022, 55(23): 4717-4727.
[4] WANG Yang,WANG WenHui,TONG Wei,JIA XiaoHui,DU YanMin. Quality Analysis of Frozen Pear Based on Color, Aroma, Taste and Texture [J]. Scientia Agricultura Sinica, 2021, 54(9): 1981-1992.
[5] XuXian XUAN,ZiLu SHENG,ZhenQiang XIE,YuQing HUANG,PeiJie GONG,Chuan ZHANG,Ting ZHENG,Chen WANG,JingGui FANG. Function Analysis of vvi-miR172s and Their Target Genes Response to Gibberellin Regulation of Grape Berry Development [J]. Scientia Agricultura Sinica, 2021, 54(6): 1199-1217.
[6] WANG YaHui, LIU XiaoHong, YONG MingLi, XIONG AiSheng, SU XiaoJun. Analysis of Changes in Phenolic Acids of Luffa cylindrica Pulp During Browning Based on Metabolomics [J]. Scientia Agricultura Sinica, 2021, 54(22): 4869-4879.
[7] YUE YingXiao,HE JinGang,ZHAO JiangLi,YAN ZiRu,CHENG YuDou,WU XiaoQi,WANG YongXia,GUAN JunFeng. Comparison Analysis on Volatile Compound and Related Gene Expression in Yali Pear During Cellar and Cold Storage Condition [J]. Scientia Agricultura Sinica, 2021, 54(21): 4635-4649.
[8] 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.
[9] CHEN Chen,JIANG AiLi,LIU ChengHui,ZHAO QiQi,ZHANG YanHui,HU WenZhong. Effect of UV-C on the Browning of Fresh-Cut Huangguan Pear [J]. Scientia Agricultura Sinica, 2020, 53(24): 5081-5090.
[10] DU YanMin,WANG WenHui,JIA XiaoHui,TONG Wei,WANG Yang,ZHANG XinNan. The Effects of Different Oxygen Concentration on Postharvest Physiology and Storage Quality of Yali Pear [J]. Scientia Agricultura Sinica, 2020, 53(23): 4918-4928.
[11] CHENG YuDou,ZHANG YaGuang,GUAN JunFeng,FENG YunXiao,HE JinGang. Effects of 1-MCP and Delayed Cold-Storage on Quality and Expression of Softening Related Genes in ‘Doyenne du Comice’ Pear During Shelf-Life [J]. Scientia Agricultura Sinica, 2020, 53(22): 4658-4666.
[12] WANG Yang, JIA XiaoHui, WANG WenHui, TONG Wei, WANG ZhiHua, DU YanMin. Quality Evaluation System Established for Pear Processed by Freezing in China [J]. Scientia Agricultura Sinica, 2019, 52(12): 2151-2160.
[13] LIN KaiLi, HUANG Qi, HUANG QiHui, JIN ZhenLiang, JIANG TianJia, ZHENG XiaoLin. Browning Inhibition and Energy Metabolism Mechanism of  Agaricus bisporus by Ergothioneine Treatment [J]. Scientia Agricultura Sinica, 2018, 51(8): 1568-1576.
[14] ZHANG Fang,WEI ZhiSheng,WANG Peng,LI KaiXuan,ZHAN Ping,TIAN HongLei. Using Neural Network Coupled Genetic Algorithm to Optimize the SPME Conditions of Volatile Compounds in Korla Pear [J]. Scientia Agricultura Sinica, 2018, 51(23): 4535-4547.
[15] OU ChunQing, JIANG ShuLing, WANG Fei, ZHAO YaNan. Genome-Wide Identification and Expression Analysis of Auxin Response Factor (ARF) Gene Family in Pear [J]. Scientia Agricultura Sinica, 2018, 51(2): 327-340.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd