基于转录组分析鉴定苹果茉莉素响应基因

doi:10.11983/CBB18235

摘要/Abstract

摘要： 为阐明外源茉莉酸甲酯(MeJA)诱导的苹果(Malus domestica)抗病分子机制, 以生长30天的Gala组培苗为试材, 用100 μmol∙L ^-1MeJA处理叶片12小时, 通过转录组测序, 结合生物信息学分析鉴定出苹果叶片中受MeJA诱导表达的基因。结果表明, 外源MeJA主要影响苹果叶片倍半萜类、三萜和类黄酮的生物合成, 以及芸薹素(BR)信号转导途径间接诱导的抗病性; 倍半萜类、三萜及类黄酮生物合成途径中的关键基因为MDP0000702120和MDP0000692178; MDP0000123379是联系芸薹素信号转导途径和植物-病原菌互作途径的关键调控基因。

关键词: 茉莉素, 抗病性, 信号转导, 转录组, 苹果

Abstract: To study the molecular mechanism of resistance induced by exogenous methyljasmonate (MeJA) in apple, tissue cultured seedlings of Gala grown for 30 days were used as materials. The leaves were treated with 100 μmol∙L ^-1 MeJA for 12 h. RNA sequencing and bioinformatics analysis were conducted to identify differentially expressed genes induced by MeJA. Exogenous MeJA mainly affected the biosynthesis of sesquiterpenoids, triterpenoids and flavonoids in apple leaves and induced disease resistance by brassinolide signal transduction pathways indirectly. The key genes regulating sesquiterpenoid and triterpenoid biosynthesis and flavonoids biosynthesis are MDP0000702120 and MDP0000692178; MDP0000123379 was the key regulatory gene linked to the brassinosteroid signaling pathway and plant-pathogen interaction pathway.

Key words: jasmonates, disease resistance, signal transduction, transcriptome, apple

张娜,刘秀霞,陈学森,吴树敬. 基于转录组分析鉴定苹果茉莉素响应基因. 植物学报, 2019, 54(6): 733-743.

Na Zhang,Xiuxia Liu,Xuesen Chen,Shujing Wu. Identifying Genes Responsive to Jasmonates in Apple Based on Transcriptome Analysis. Chinese Bulletin of Botany, 2019, 54(6): 733-743.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB18235

https://www.chinbullbotany.com/CN/Y2019/V54/I6/733

图/表 9

参考文献 22

[1]	宾金华, 潘瑞炽 ( 1995). 茉莉酸甲酯的生理生化及在植物抗病中的作用. 植物学通报 12(4), 17-21.
[2]	陈学森, 郭文武, 徐娟, 从佩华, 王力荣, 刘崇怀, 李秀根, 吴树敬, 姚玉新, 陈晓流 ( 2015). 主要果树果实品质遗传改良与提升实践. 中国农业科学 48, 3524-3540.
[3]	弓德强, 谷会, 张鲁斌, 洪克前, 朱世江 ( 2013). 杧果采前喷施茉莉酸甲酯对其抗病性和采后品质的影响. 园艺学报 40, 49-57.
[4]	国立耘, 李金云, 李保华, 张新忠, 周增强, 李广旭, 王英姿, 李晓军, 黄丽丽, 孙广宇, 文耀东 ( 2009). 中国苹果枝干轮纹病发生和防治情况. 植物保护 35(4), 120-123.
[5]	李灿婴, 葛永红, 朱丹实, 张惠君 ( 2015). 采后茉莉酸甲酯处理对富士苹果青霉病和贮藏品质的影响. 食品科学 36(2), 255-259.
[6]	李梦莎, 阎秀峰 ( 2014). 植物的环境信号分子茉莉酸及其生物学功能. 生态学报 34, 6779-6788.
[7]	刘志, 张华磊, 谢兴斌, 束怀瑞, 郝玉金 ( 2009). 茉莉酸甲酯诱导的苹果悬浮细胞的防卫响应. 园艺学报 36, 171-178.
[8]	罗昌国, 袁启凤, 裴晓红, 吴亚维, 郑伟, 章镇 ( 2013). 富士苹果MdWRKY40b基因克隆及其对白粉病的抗性分析. 西北植物学报 33, 2382-2387.
[9]	麻宝成, 朱世江 ( 2006). 苯丙噻重氮和茉莉酸甲酯对采后香蕉果实抗病性及相关酶活性的影响. 中国农业科学 39, 1220-1227.
[10]	汪开拓, 郑永华, 唐文才, 李廷君, 张卿, 尚海涛 ( 2012). 茉莉酸甲酯处理对葡萄果实NO和H₂O₂水平及植保素合成的影响. 园艺学报 39, 1559-1566.
[11]	王英珍, 程瑞, 张绍铃, 白彬, 何子顺, 张虎平 ( 2016). 采前茉莉酸甲酯(MeJA)处理对梨果实抗病性的影响. 果树学报 33, 694-700.
[12]	许晴晴, 郜海燕, 陈杭君 ( 2014). 茉莉酸甲酯对蓝莓贮藏品质及抗病相关酶活性的影响. 核农学报 28, 1226-1231.
[13]	张芮 ( 2015). 苹果MdWRKY33基因在轮纹病抗性形成中的作用机制研究. 博士论文. 泰安: 山东农业大学. pp. 70-73.
[14]	Chen XK, Zhang JY, Zhang Z, Du XL, Du BB, Qu SC ( 2012). Overexpressing MhNPR1 in transgenic Fuji apples enhances resistance to apple powdery mildew. Mol Biol Rep 39, 8083-8089.
[15]	Langmead B, Trapnell C, Pop M, Salzberg SL ( 2009). Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10, R25.
[16]	Li H, Durbin R ( 2009). Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25, 1754-1760.
[17]	Sun X, Matus JT, Wong DCJ, Wang Z, Chai F, Zhang L, FangT, Zhao L, Wang Y, Han Y, Wang Q, Li S, Liang Z, Xin H ( 2018). The GARP/MYB-related grape transcription factor AQUILO improves cold tolerance and promotes the accumulation of raffinose family oligosaccharides. J Exp Bot 69, 1749-1764.
[18]	Velasco R, Zharkikh A, Affourtit J, Dhingra A, Cestaro A, Kalyanaraman A, Fontana P, Bhatnagar SK, Troggio M, Pruss D, Salvi S, Pindo M, Baldi P, Castelletti S, Cavaiuolo M, Coppola G, Costa F, Cova V, Dal Ri A, Goremykin V, Komjanc M, Longhi S, Magnago P, Malacarne G, Malnoy M, Micheletti D, Moretto M, Perazzolli M, Si-Ammour A, Vezzulli S, Zini E, Eldredge G, Fitzgerald LM, Gutin N, Lanchbury J, Macalma T, Mitchell JT, Reid J, Wardell B, Kodira C, Chen ZT, Desany B, Niazi F, Palmer M, Koepke T, Jiwan D, Schaeffer S, Krishnan V, Wu CJ, Chu VT, King ST, Vick J, Tao QZ, Mraz A, Stormo A, Stormo K, Bogden R, Ederle D, Stella A, Vecchietti A, Kater MM, Masiero S, Lasserre P, Lespinasse Y, Allan AC, Bus V, Chagné D, Crowhurst RN, Gleave AP, Lavezzo E, Fawcett JA, Proost S, Rouzé P, Sterck L, Toppo S, Lazzari B, Hellens RP, Durel CE, Gutin A, Bumgarner RE, Gardiner SE, Skolnick M, Egholm M, Van de Peer Y, Salamini F, Viola R ( 2010). The genome of the domesticated apple ( Malus × domestica Borkh.). Nat Genet 42, 833-839.
[19]	Xue YK, Shui GH, Wenk MR ( 2014). TPS1 drug design for rice blast disease in Magnaporthe oryzae. Springer Plus 3, 18.
[20]	Ye J, Fang L, Zheng HK, Zhang Y, Chen J, Zhang ZJ, Wang J, Li ST, Li RQ, Bolund L, Wang J ( 2006). WEGO: a web tool for plotting GO annotations. Nucleic Acids Res 34, W293-W297.
[21]	Zhang HY, Ma LC, Turner M, Xu HX, Dong Y, Jiang S ( 2009). Methyl jasmonate enhances biocontrol efficacy of Rhodotorula glutinis to postharvest blue mold decay of pears. Food Chem 117, 621-626.
[22]	Zhang Y, Shi XP, Li BH, Zhang QM, Liang WX, Wang CX ( 2016). Salicylic acid confers enhanced resistance to Glomerella leaf spot in apple. Plant Physiol Biochem 106, 64-72.

Sample	Total reads	Total base pairs	Total mapped reads	Perfect match	Mismatch	Unique match	Muti-position match	Total unmapped reads
CTRL1	15336268 (100.00%)	751477132 (100.00%)	10932142 (71.28%)	6783455 (44.23%)	4148687 (27.05%)	9149425 (59.66%)	1782717 (11.62%)	4404126 (28.72%)
MeJA	15339040 (100.00%)	751612960 (100.00%)	11052987 (72.06%)	6921067 (45.12%)	4131920 (26.94%)	9177064 (59.83%)	1875923 (12.23%)	4286053 (27.94%)

Sample	Total reads	Total base pairs	Total mapped reads	Perfect match	Mismatch	Unique match	Muti-position match	Total unmapped reads
CTRL1	15336268 (100.00%)	751477132 (100.00%)	10932142 (71.28%)	6783455 (44.23%)	4148687 (27.05%)	9149425 (59.66%)	1782717 (11.62%)	4404126 (28.72%)
MeJA	15339040 (100.00%)	751612960 (100.00%)	11052987 (72.06%)	6921067 (45.12%)	4131920 (26.94%)	9177064 (59.83%)	1875923 (12.23%)	4286053 (27.94%)

Gene ID	Log₂ ratio (MeJA/CTRL1)	KEGG orthology	Blast nr	Pathway
MDP0000702120	12.32	K14181	(-)-alpha-pinene synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000205617	7.28	K14181	Putative pinene synthase	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000919962	6.57	K14181	(-)-alpha-pinene synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000225361	3.72	K14181	(-)-germacrene D synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000248152	3.24	K14181	(-)-germacrene D synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000692178	12.12	K00517	Cytochrome P450 82G1-like	Flavonoid biosynthesis
MDP0000503940	4.67	K05277	Protein SRG1-like	Flavonoid biosynthesis
MDP0000225939	3.55	K05277	Flavonol synthase/flavanone 3-hydroxylase-like	Flavonoid biosynthesis
MDP0000523477	-3.17	K01859	Chalcone-flavonone isomerase-like isoform X5	Flavonoid biosynthesis
MDP0000317152	13.53	K13407	Cytochrome P450 94C1-like	Cutin, suberine and wax biosynthesis
MDP0000153822	3.25	K13407	Cytochrome P450 94B3-like	Cutin, suberine and wax biosynthesis
MDP0000218691	3.89	K01183	Acidic endochitinase-like	Amino sugar and nucleotide sugar metabolism
MDP0000280265	3.22	K01183	Acidic endochitinase-like	Amino sugar and nucleotide sugar metabolism
MDP0000687619	12.96	K09755	Cytochrome P450 CYP736A12-like	Phenylpropanoid biosynthesis
MDP0000539956	3.9	K09755	Cytochrome P450 CYP736A12-like	Phenylpropanoid biosynthesis
MDP0000313394	6.44	K13267	Cytochrome P450 CYP736A12-like	Isoflavonoid biosynthesis
MDP0000813805	-6.29	K09840	9-cis-epoxycarotenoid dioxygenase NCED3, chloroplastic-like	Carotenoid biosynthesis
MDP0000228070	-5.24	K09840	9-cis-epoxycarotenoid dioxygenase NCED1, chloroplastic-like	Carotenoid biosynthesis

Gene ID	Log₂ ratio (MeJA/CTRL1)	KEGG orthology	Blast nr	Pathway
MDP0000702120	12.32	K14181	(-)-alpha-pinene synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000205617	7.28	K14181	Putative pinene synthase	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000919962	6.57	K14181	(-)-alpha-pinene synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000225361	3.72	K14181	(-)-germacrene D synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000248152	3.24	K14181	(-)-germacrene D synthase-like	Sesquiterpenoid and triterpenoid biosynthesis
MDP0000692178	12.12	K00517	Cytochrome P450 82G1-like	Flavonoid biosynthesis
MDP0000503940	4.67	K05277	Protein SRG1-like	Flavonoid biosynthesis
MDP0000225939	3.55	K05277	Flavonol synthase/flavanone 3-hydroxylase-like	Flavonoid biosynthesis
MDP0000523477	-3.17	K01859	Chalcone-flavonone isomerase-like isoform X5	Flavonoid biosynthesis
MDP0000317152	13.53	K13407	Cytochrome P450 94C1-like	Cutin, suberine and wax biosynthesis
MDP0000153822	3.25	K13407	Cytochrome P450 94B3-like	Cutin, suberine and wax biosynthesis
MDP0000218691	3.89	K01183	Acidic endochitinase-like	Amino sugar and nucleotide sugar metabolism
MDP0000280265	3.22	K01183	Acidic endochitinase-like	Amino sugar and nucleotide sugar metabolism
MDP0000687619	12.96	K09755	Cytochrome P450 CYP736A12-like	Phenylpropanoid biosynthesis
MDP0000539956	3.9	K09755	Cytochrome P450 CYP736A12-like	Phenylpropanoid biosynthesis
MDP0000313394	6.44	K13267	Cytochrome P450 CYP736A12-like	Isoflavonoid biosynthesis
MDP0000813805	-6.29	K09840	9-cis-epoxycarotenoid dioxygenase NCED3, chloroplastic-like	Carotenoid biosynthesis
MDP0000228070	-5.24	K09840	9-cis-epoxycarotenoid dioxygenase NCED1, chloroplastic-like	Carotenoid biosynthesis

Plant hormone signaling pathway	Gene ID	Log₂ ratio (MeJA/CTRL1)
Auxin	MDP0000300452	-1.4
	MDP0000255223 MDP0000213864	1.1 -1.1
	MDP0000143749 MDP0000221322	1.3 -1.6
	MDP0000873893 MDP0000214081	1.5 -1.1
	MDP0000786165	1.1
Cytokinine	MDP0000146863	-1.1
	MDP0000285242 MDP0000225179	1.6 -1.0
	MDP0000250737	-1.5
Gibberellin	MDP0000227827 MDP0000868088	-1.3 -1.0
Gibberellin	MDP0000284679 MDP0000227056 MDP0000397638	1.4 1.3 -2.2
Abscisic acid	MDP0000437033 MDP0000203818 MDP0000283274	-1.6 -1.3 -1.2
Ethylene	MDP0000275915	-2.4
Ethylene	MDP0000855671 MDP0000127134 MDP0000235313 MDP0000805422 MDP0000787281	3.1 2.5 1.8 1.2 1.0
Brassinosteroid	MDP0000123379 MDP0000319460 MDP0000319708 MDP0000904826 MDP0000196862	12.5 1.9 1.0 1.0 -1.8
	MDP0000217213 MDP0000897962 MDP0000290950 MDP0000620422 MDP0000303744 MDP0000189315 MDP0000741253 MDP0000223726 MDP0000189841	1.9 1.3 -2.3 -1.8 -1.6 -1.1 -1.0 -1.0 -1.0
	MDP0000130143	-12.8
	MDP0000320017	-1.4
	MDP0000135392	-1.0
Jasmonic acid	MDP0000565690 MDP0000187921 MDP0000889413 MDP0000301927	2.8 2.6 1.7 1.2
Jasmonic acid	MDP0000603546 MDP0000242554 MDP0000029168 MDP0000900024 MDP0000226497 MDP0000290263	3.6 2.2 1.6 1.6 1.0 -1.0
Salicylic acid	MDP0000292425	1.8
Salicylic acid	MDP0000529682 MDP0000375992	1.1 -4.3