[1]冯娇,王武,侯旭东,等.基于转录组测序技术探究GA3和CPPU抑制葡萄果锈产生的机理[J].江苏农业学报,2017,(04):895-903.[doi:doi:10.3969/j.issn.1000-4440.2017.04.026]
　FENG Jiao,WANG Wu,HOU Xu-dong,et al.Underlying mechanism of GA3 and CPPU inhibiting grape fruit russet discovered based on transcriptome sequencing[J].,2017,(04):895-903.[doi:doi:10.3969/j.issn.1000-4440.2017.04.026]
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基于转录组测序技术探究GA3和CPPU抑制葡萄果锈产生的机理()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年04期
页码:
895-903
栏目:
园艺
出版日期:
2017-08-30

文章信息/Info

Title:
Underlying mechanism of GA3 and CPPU inhibiting grape fruit russet discovered based on transcriptome sequencing
作者:
冯娇; 王武; 侯旭东; 陶建敏
（南京农业大学园艺学院，江苏南京210095）
Author(s):
FENG Jiao; WANG Wu; HOU Xu-dong; TAO Jian-min
(College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China)
关键词:
葡萄; 果锈; 转录组测序; 代谢通路; 差异表达基因; GA3(赤霉素); CPPU[N-(2-氯-4-吡啶基)-N′-苯基脲]
Keywords:
Shine Muscat; berry reddish-brown blotches; transcriptome sequencing; metabolism pathway; differently expressed genes; GA3; CPPU
分类号:
S663.1
DOI:
doi:10.3969/j.issn.1000-4440.2017.04.026
文献标志码:
A
摘要:
为探究GA3（赤霉素）和CPPU[N-(2-氯-4-吡啶基)-N′-苯基脲]抑制葡萄果锈生成的内在机理，以阳光玫瑰葡萄为试验材料，在盛花期用25 mg/L GA3处理花穗，花后2周用25 mg/L GA3+10 mg/L CPPU再次处理花穗，以清水处理作为对照，取果锈生成关键期的2个葡萄果皮样品，应用Illumina RNA-Seq测序系统进行转录组分析。共获得55 314 986条有效数据和2 373个差异基因，GO功能注释到细胞组分、分子功能及生物学过程3个大类30个功能组，大量基因富集于碳水化合物代谢、生物合成等生物学过程。KEGG代谢通路分析结果显示将1 186个差异表达基因注释到126条代谢通路，富集于“代谢途径”与“苯丙烷合成”代谢通路。通过qRT-PCR方法对筛选的9个差异表达基因进行验证，荧光定量结果与测序结果相符。研究结果说明GA3和CPPU处理减少葡萄果锈的产生主要与苯丙烷合成及类黄酮代谢相关，GA3和CPPU显著抑制苯丙烷及类黄酮合成相关基因表达，进而降低了果锈生成路径中相关酶的活性，减少了果锈生成相关次生代谢物的产生，抑制葡萄果锈的生成。
Abstract:
To probe into the mechanism of GA3(gibberellin A3) and CPPU(forchlorfenuron) inhibiting the tiny reddish-brown blotches on the surface of grape berries, the floral clusters of Shine Muscat grape was treated with 25 mg/L GA3 at full blossom, and 25 mg/L GA3 combined with 10 mg/L CPPU 2 weeks after full blossom. Water treatment was set as control. Two pericarp samples at critical period for fruit russet-genesis were subjected to transcriptome sequencing using Illumina RNA-Seq system. Approximately 55 314 986 clean data and 1 186 differentially expressed genes (DEGs) were identified. Gene oncology(GO) functions annotated to 30 functional groups belonging to three categories: cell components, molecular functions, and biological processes. A large amount of genes were enriched in the biological processes ‘carbohydrate metabolism’ and ‘biosynthesis’. Analysis of KEGG metabolic pathways demonstrated that most enriched EDGs were associated with ‘metabolic pathway’ and ‘phenylpropanoid synthesis’ out of 126 total pathways. QRT-PCR assay results for 9 selected DEGs showed that the expression levels of 9 genes were consistent with the data obtained by RNA-seq. It was indicated that GA3 and CPPU treatment might have inhibited the gene expression and enzyme activity of ‘phenylpropanoid biosynthesis’ and ‘flavonoid metabolism’ pathways which in turn reduced the production of secondary metabolites, the precursors of reddish-brown blotches, thereby inhibiting the skin browning of grape fruits.

参考文献/References:

[1]KHALILURREHMAN M, SUN L, LI C X, et al. Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape[J]. BMC Plant Biology, 2017, 17 (1): 18.
[2]周朝华，刘金凤.黄金梨果锈发生严重的原因及预防措施[J].落叶果树，2005, 37(5)：37-39.
[3]滕元文，沈玉英，周先. 砂梨果皮锈斑成因及解决对策[J]. 中国南方果树, 2005, 34 (3)：52-54.
[4]OGORO A, ONO T, KAGAMI H, et al. Skin-browning occurred on berries of ‘Suihou’ grape[J]. Bull Okayama Agri Exp Sta, 2007, 25:5-10.
[5]李亚锋. 关于金冠苹果果锈形成原因的研究[J]. 吉林农业,2010(9)：66.
[6]聂继云，重雅凤，马智勇，等. 金冠苹果果锈的发生原因及防治[J]. 北方果树,2001(3)：3-4.
[7]李健花，高晶晶，冯新新，等. ‘金冠’苹果与其无锈芽变的果皮性状比较和防锈技术研究[J]. 园艺学报, 2014,41(1): 35-43.
[8]YUKA S, KEISUKE M, HIROYUKI I, et al. Skin browning and expression of PPO, STS, and CHS genes in the grape berries of ‘Shine Muscat’ [J]. Journal Japanese Society for Horticultural Science, 2014, 83(2): 122-132.
[9]WANG Y Z, DAI M S, CAI D Y, et al. A review for the molecular research of russet/semi-russet of sand pear exocarp and their genetic characters [J]. Scientia Horticulturae, 2016, 210: 138-142.
[10]马克元，程福厚，傅玉瑚，等. 鸭梨果实果点和锈斑的发育[J]. 园艺学报,1995, 22（3）：295-296.
[11]郑永华，李三玉，席芳. 枇杷冷藏过程中果肉木质化与细胞壁物质变化的关系[J]. 植物生理学报,2000, 26 (4)：306-310.
[12]RETAMALES J, BANGERTH F, COOPER T, et al. Effects of CPPU and GA3 on fruit quality of ‘Sultanina’ table grape [J]. Plant Bioregulators in Horticulture, 1995, 394: 149-158.
[13]辛守鹏，刘帅，余阳，等.赤霉素与细胞分裂素对葡萄果实邻近叶光合特性及果实品质的影响[J]. 应用生态学报, 2015,26(7): 1814-1820.
[14]顾克余，周蓓蓓，宋长年，等．植物生长调节剂及其在葡萄生产上的应用综述［J］．江苏农业科学，2015,43(7):13-16.
[15]陈国品，白先进，文仁德, 等. 赤霉素对延后栽培酿酒葡萄西拉生长发育及果实品质的影响[J]. 南方农业学报, 2016，47(5)：694-699.
[16]吴俊，钟家煌，徐凯，等. 外源GA3对藤稔葡萄果实生长发育及内源激素水平的影响[J]. 果树学报, 2001,18(4): 209-212.
[17]夏瑾华，喻慧荣，邹如意，等．6-BA、赤霉素和水杨酸对甜瓜种子萌发的影响[J]．江苏农业科学，2015，43(12)：204-206.
[18]张溢，袁雅珍，余露霞，等. 植物生长调节剂对月季切花保鲜效果的影响[J]. 南方农业学报, 2016，47(3)：454-458.
[19]刘耀光，曹慧. CPPU对红地球葡萄果实主要经济性状的影响[J].山西果树,2007(2)：3-4.
[20]侯玉茹，王宝刚，冯晓元，等. CPPU和GA3在葡萄中的残留动态及对果实品质的影响[J]. 果树学报，2012，29（1）：36-41.
[21]余智莹，张萌，陶建敏. GA3和CPPU对凉玉葡萄果实品质的影响[J]. 中外葡萄与葡萄酒, 2010(7)：49-51.
[22]CHEN G, WANG C, SHI T L. Overview of available methods for diverse RNA-Seq data analyses[J]. Science China Life Sciences, 2011, 54 (12): 1121-1128.
[23]冯超，朱长青，徐昌杰，等. RNA-Seq在果树学研究中的应用[J]. 果树学报,2016, 31（01）：115-124.
[24]FOLTA K M, CLANCY M A, CHAMALA S, et al. A transcript accounting from diverse tissues of a cultivated strawberry[J]. Plant Genome ,2010, 3 (2): 90-105.
[25]ZHANG J Z, AI X Y, SUN L M, et al. Transcriptome profile analysis of flowering molecular processes of early flowering trifoliate orange mutant and the wild-type [Poncirus trifoliata (L.) Raf.] by massively parallel signature sequencing[J]. BMC Genomics, 2011,12 (1): 63.
[26]D HONT A, DENOEUD F, AURY J M, et al. The banana (Musa acuminata) genome and the evolution of monocotyledonous plants[J]. Nature, 2012, 488 (7410): 213-217.
[27]FENG C, CHEN M, XU C J, et al. Transcriptomic analysis of Chinese bayberry ( Myrica rubra ) fruit development and ripening using RNA-Seq[J]. BMC Genomics, 2012, 13 (1): 19.
[28]WU J, WANG Z, SHI Z, et al. The genome of pear (Pyrus bretschneideri Rehd.)[J]. Genome Research, 2012, 23 (2): 396-408.
[29]ZHANG Y, ZHU J, DAI H. Characterization of transcriptional differences between columnar and standard apple trees using RNA-Seq[J]. Plant Molecular Biology Reporter, 2012, 30 (4): 957-965.
[30]LUCA V, ALBERTO F, SARA Z, et al. De novo transcriptome characterization of Vitis vinifera cv. Corvina unveils varietal diversity[J]. BMC Genomics , 2013,14 (1): 41.
[31]王云生，聂飞，林顺权. 果树高通量测序的最新研究进展[J]. 基因组学与应用生物学,2015, 34（9）：2034-2043.
[32]殷向静. 山葡萄果实转录组分析及白藜芦醇调控机理研究[D]. 咸阳:西北农林科技大学,2016.
[33]李锡花，吴嫚，于霁雯，等. 棉花纤维发育早期RNA-Seq转录组分析[J]. 棉花学报, 2013,25 (3)：189-196.
[34]张少平，洪建基，邱珊莲，等. 紫背天葵高通量转录组测序分析[J]. 园艺学报, 2016, 43 (5): 935-946.
[35]吴锦程，唐朝晖，陈群，等.不同贮藏温度对枇杷果肉木质化及相关酶活性的影响[J].武汉植物学研究，2006,24(3): 235-239.
[36]HENG W, LIU L, WANG M D, et al. Differentially expressed genes related to the formation of russet fruit skin in a mutant of ‘Dangshansuli’ pear (Pyrus bretchnederi Rehd.) determined by suppression subtractive hybridization [J]. Euphytica, 2014, 196(2): 285-297.
[37]欧阳光察，薛应龙. 植物苯丙烷类代谢的生理意义及其调控[J]. 植物生理学通讯,1988（3）：9-16.
[38]WANG Y Z, DAI M S, ZHANG S J, et al. Exploring candidate genes for pericarp russet pigmentation of sand pear (Pyrus pyrifolia) via RNA-Seq data in two genotypes contra sting for pericarp color[J]. PLoS One, 2014, 9(1):e83675.
[39]WANG Y Z, ZHANG S, DAI M S, et al. Pigmentation in sand pear (Pyrus pyrifolia) fruit: biochemical characterization, gene discovery and expression analysis with exocarp pigmentation mutant[J]. Plant Mol Biol , 2014, 85:123-134.
[40]LEGAY S, GUERRIERO G, DELERUELLE A, et al. Apple russeting as seen through the RNA-Seq lens: strong alterations in the exocarp cell wall [J]. Plant Mol Biol, 2015, 88:21-40.
[41]KOBAYASHI S, ISHIMARU M, DING C K, et al. Comparison of UDP-glucose: flavornoid 3-O-glucosyltransferase (UFGT) gene sequences between white grapes (Vitis vinifera) and their sports with red skin [J]. Plant Science, 2001, 160(3): 543-550.
[42]王莉，史玲玲，张艳霞，等. 植物次生代谢物途径及其研究进展[J]. 武汉植物学研究,2007,25(5)：500-508.
[43]秦秋琳. 银杏中与木质素合成和苯丙氨酸代谢相关的转录调控因子的克隆与研究[D]. 上海:复旦大学，2007.
[44]张宽朝，金青，蔡永萍，等. 苯丙氨酸解氨酶与其重要次生代谢产物调控中的作用研究进展[J]. 中国农学通报,2008,24(12):59-62.

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备注/Memo

备注/Memo:
收稿日期：2017-04-06 基金项目：国家现代农业葡萄产业技术体系项目(CARS-30)； 国家“948”重点项目(2016-X19)；江苏省科技厅设施条件下葡萄高效栽培新技术研究与应用项目（BE2014326） 作者简介：冯娇（1992-），女，山西运城人，硕士，主要从事果树生理与分子技术研究。(E-mail)478710492@qq.com 通讯作者：陶建敏，(E-mail)tjm266@sina.com
更新日期/Last Update: 2017-09-01