转录组与代谢组联合分析菠萝网纱覆盖防寒机制

doi:10.13560/j.cnki.biotech.bull.1985.2022-0203

生物技术通报 ›› 2022, Vol. 38 ›› Issue (11): 58-69.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0203

转录组与代谢组联合分析菠萝网纱覆盖防寒机制

刘传和¹(), 贺涵¹, 何秀古²(), 赖秋勤¹, 刘开¹, 邵雪花¹, 赖多¹, 匡石滋¹, 肖维强¹

1.广东省农业科学院果树研究所农业农村部南亚热带果树生物学与遗传资源利用重点实验室广东省热带亚热带果树研究重点实验室，广州 510640
2.广东省农业科学院，广州 510640

收稿日期:2022-02-21 出版日期:2022-11-26 发布日期:2022-12-01
作者简介:刘传和，男，博士，副研究员，研究方向：菠萝优质高效种植技术与新品种选育；E-mail：founderlch@126.com
基金资助:
乡村振兴战略专项资金(403-2018-XMZC-0002-90);乡村振兴战略专项资金(TS-3-1);国家重点研发计划项目(2019YFD1001100);广东省现代农业产业技术体系创新团队建设专项资金(2021KJ109);广东省现代农业产业技术体系创新团队建设专项资金(2022KJ109);广东省农业科学院菠萝产业研究院项目，广东省农村科技特派员项目(KTP20210395)

Unveiling the Mechanisms of Pineapple Responding to Anti-chilling by Gauze Covering in Winter via Transcriptome and Metabolome Profiling

LIU Chuan-he¹(), HE Han¹, HE Xiu-gu²(), LAI Qiu-qin¹, LIU Kai¹, SHAO Xue-hua¹, LAI Duo¹, KUANG Shi-zi¹, XIAO Wei-qiang¹

1. Institute of Fruit Tree Research，Guangdong Academy of Agricultural Sciences/Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization Ministry of Agriculture and Rural Affairs/ Guangdong Provincial Key Laboratory of Tropical and Subtropical Fruit Tree Research，Guangzhou 510640
2. Guangdong Academy of Agricultural Sciences，Guangzhou 510640

Received:2022-02-21 Published:2022-11-26 Online:2022-12-01

摘要/Abstract

摘要：

本研究以不覆盖为对照（CK），通过转录组、代谢组进行基因表达和代谢物差异分析，探究冬季菠萝黑色网纱覆盖防寒（FH）的生理及分子机制。转录组分析表明，与CK相比，FH中筛选到1 125个差异表达基因，其中表达上调和下调基因分别为417个和708个。差异基因主要富集在淀粉和蔗糖代谢、内质网蛋白加工、苯丙素生物合成和氨基酸生物合成等通路；筛选出19个与防寒相关的候选关键差异基因，包括WRKY、MYB、bHLH等转录因子基因及ERD6-like、β-葡萄糖苷酶等酶基因。代谢组分析表明，与CK相比，FH中筛选到151个差异代谢物，包括含量上调57个和含量下调94个。差异代谢物主要富集在嘌呤和嘧啶代谢、氨基酸类代谢、脂类代谢和黄酮类代谢等通路；筛选出26个与防寒相关的关键差异代谢物，主要包括类黄酮、氨基酸、多胺、脂和糖类等。本研究结果表明，黑色网纱覆盖通过影响菠萝相关基因差异表达和代谢物变化来调控菠萝生长发育，安全越冬，为今后指导菠萝冬季防寒，挖掘菠萝耐寒功能基因奠定基础。

关键词: 菠萝, 网纱覆盖, 防寒, 差异表达基因, 差异代谢物

Abstract:

The aim of this trial is to investigate the physiological and molecular mechanisms of pineapple responding to anti-chilling by gauze covering practices（FH）through the gene expression and metabolite differences analysis of transcriptomics and metabolomics compared with no covering control（CK）. The results of transcriptomics indicated that 1 125 differential expressed genes were detected in FH，including 417 up-regulated genes and 708 down-regulated genes，when compared with CK. The differential expressed genes were mainly enriched in 4 KEGG pathways including starch and sucrose metabolism，protein processing in endoplasmic reticulum，phenylpropanoid biosynthesis and amino acid biosynthesis. Nineteen differential expressed genes were found to be related to cold resistance，including a few of transcription factors named WRKY，MYB and bHLH as well as some enzyme genes named ERD6-like and β-glucosidase. Metabolome profiling suggested that a total of 151 differential metabolites were detected in FH group，including 57 and 94 up-regulated and down-regulated ones when compared with CK，respectively. The differential metabolites were mainly enriched in the 4 metabolic pathways including purine and pyrimidine metabolism，amino acid metabolism，lipids metabolism and flavonoid biosynthesis. A total of 26 differential metabolites were found to be involved in cold resistance，including flavonoids，amino acids，polyamine，lipids and carbohydrates. The results of this study reveal that that pineapple growth and safe overwintering may be regulated by affecting its differential gene expression and metabolite changes while using black gauze covering，which lays a foundation in guiding the pineapple anti-chilling in winter and digging the function genes of pineapple resistant to cold.

Key words: pineapple, gauze covering, anti-chilling, differential expressed genes, differential metabolites

刘传和, 贺涵, 何秀古, 赖秋勤, 刘开, 邵雪花, 赖多, 匡石滋, 肖维强. 转录组与代谢组联合分析菠萝网纱覆盖防寒机制[J]. 生物技术通报, 2022, 38(11): 58-69.

LIU Chuan-he, HE Han, HE Xiu-gu, LAI Qiu-qin, LIU Kai, SHAO Xue-hua, LAI Duo, KUANG Shi-zi, XIAO Wei-qiang. Unveiling the Mechanisms of Pineapple Responding to Anti-chilling by Gauze Covering in Winter via Transcriptome and Metabolome Profiling[J]. Biotechnology Bulletin, 2022, 38(11): 58-69.

图/表 10

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样品Sample	CK	FH
SNP数量SNP Number	209 800	215 075
基因区SNP数量Genic SNP	191 018（91.05%）	196 217（91.23%）
基因间区SNP数量 Intergenic SNP	18 782（8.95%）	18 858（8.77%）
杂合型Heterozygosity	61.67%	61.51%
转换Transition	65.44%	65.40%
颠换Transversion	34.56%	34.60%
转换/颠换 Transition/Transversion	1.89	1.89

样品Sample	CK	FH
SNP数量SNP Number	209 800	215 075
基因区SNP数量Genic SNP	191 018（91.05%）	196 217（91.23%）
基因间区SNP数量 Intergenic SNP	18 782（8.95%）	18 858（8.77%）
杂合型Heterozygosity	61.67%	61.51%
转换Transition	65.44%	65.40%
颠换Transversion	34.56%	34.60%
转换/颠换 Transition/Transversion	1.89	1.89

基因ID Gene ID	基因注释Annotation	差异倍数log₂FC	参考文献Reference
LOC109722013	WRKY75	1.64	低温胁迫下诱导上调表达^[13]
LOC109719592	WRKY24-like	1.63	抗寒、促进植株生长^[14]
LOC109722010	bHLH41（basic helix-loop-helix）	2.14	低温、ABA、盐胁迫响应^[15]
LOC109708332	bHLH111-like	1.46	花青素生物合成^[16]
LOC109707663	MYB-like protein AA	1.62	抗寒^[17]
LOC109723346	MYB44-like	1.48	参与MAPK级联反应，增强渗透压耐受性^[18]
LOC109713884	NAC35	1.96	参与植物生物与非生物逆境应答^[19]
LOC109703579	NAC94	2.37	启动子部位具有与低温胁迫相关的调控元件^[20]
LOC109727211	多聚半乳糖醛酸酶Polygalacturonase	-2.59	参与果胶的降解^[21]
LOC109711455	β-葡萄糖苷酶β -glucosidase	2.50	海巴戟抗寒相关^[22]
LOC109712027	硝酸还原酶Nitrate reductase[NADH]-like	3.68	调节植物氮代谢与非生物胁迫^[23]
LOC109704466	富含半胱氨酸类受体蛋白激酶 Cysteine-rich receptor-like protein kinase	2.72	感知胁迫信号和诱发免疫应答^[24]
LOC109708872	丝氨酸/苏氨酸蛋白激酶PBL18 Serine/threonine-protein kinase PBL18	2.67	调节拟南芥的免疫应答^[25]
LOC109726085	水通道蛋白NIP2-2 like（Nodulin-intrinsic protein）	1.18	毛竹抗逆性调控^[26]
LOC109711392	ERF014（ethylene-responsive transcription factor）	1.34	植物免疫调节^[27]
LOC109707256	AP2/ERF（APETALA2/ Ethylene-responsive transcription factor）	1.53	逆境调控^[28]
LOC109709179	ERD6-like（early response to dehydration 6-like）	-2.40	糖运输^[29]
LOC109727398	HSP83（Heat shock protein）	1.65	抵抗高温^[30]
LOC109706855	谷氨酸受体Glutamate receptor	2.55	应答多种生物和非生物环境胁迫^[31]

基因ID Gene ID	基因注释Annotation	差异倍数log₂FC	参考文献Reference
LOC109722013	WRKY75	1.64	低温胁迫下诱导上调表达^[13]
LOC109719592	WRKY24-like	1.63	抗寒、促进植株生长^[14]
LOC109722010	bHLH41（basic helix-loop-helix）	2.14	低温、ABA、盐胁迫响应^[15]
LOC109708332	bHLH111-like	1.46	花青素生物合成^[16]
LOC109707663	MYB-like protein AA	1.62	抗寒^[17]
LOC109723346	MYB44-like	1.48	参与MAPK级联反应，增强渗透压耐受性^[18]
LOC109713884	NAC35	1.96	参与植物生物与非生物逆境应答^[19]
LOC109703579	NAC94	2.37	启动子部位具有与低温胁迫相关的调控元件^[20]
LOC109727211	多聚半乳糖醛酸酶Polygalacturonase	-2.59	参与果胶的降解^[21]
LOC109711455	β-葡萄糖苷酶β -glucosidase	2.50	海巴戟抗寒相关^[22]
LOC109712027	硝酸还原酶Nitrate reductase[NADH]-like	3.68	调节植物氮代谢与非生物胁迫^[23]
LOC109704466	富含半胱氨酸类受体蛋白激酶 Cysteine-rich receptor-like protein kinase	2.72	感知胁迫信号和诱发免疫应答^[24]
LOC109708872	丝氨酸/苏氨酸蛋白激酶PBL18 Serine/threonine-protein kinase PBL18	2.67	调节拟南芥的免疫应答^[25]
LOC109726085	水通道蛋白NIP2-2 like（Nodulin-intrinsic protein）	1.18	毛竹抗逆性调控^[26]
LOC109711392	ERF014（ethylene-responsive transcription factor）	1.34	植物免疫调节^[27]
LOC109707256	AP2/ERF（APETALA2/ Ethylene-responsive transcription factor）	1.53	逆境调控^[28]
LOC109709179	ERD6-like（early response to dehydration 6-like）	-2.40	糖运输^[29]
LOC109727398	HSP83（Heat shock protein）	1.65	抵抗高温^[30]
LOC109706855	谷氨酸受体Glutamate receptor	2.55	应答多种生物和非生物环境胁迫^[31]

代谢物ID Metabolites ID	名称 Compound	分类 Classification	差异倍数 log₂FC	P值 P-value	VIP值 VIP
mws0064	圣草酚Eriodictyol	多酚黄酮Polyphenol flavonoid	14.07	0.0040	1.31
pme0376	柚皮素Naringenin	黄酮Flavonoid	10.31	0.023607	1.26
pme2960	柚皮素查尔酮Naringenin chalcone	黄酮Flavonoid	9.64	0.010187	1. 30
mws0914	短叶松素 Pinobanksin	黄酮Flavonoid	10.29	0.00147	1.32
pmn001668	芹菜素-3-邻鼠李糖苷Pigenin-3-O-rhamnoside	黄酮糖苷Flavone glycoside	1.61	0.0064	1.30
Lmtn002796	香橙素-7-葡萄糖苷Aromadendrin 7-glucoside	黄酮糖苷Flavone glycoside	1.15	0.0214	1.20
HJN041	表儿茶素葡萄糖苷Epicatechin glucoside	黄酮糖苷Flavone glycoside	10.18	0.0027	1.32
mws0712	N-丙酰甘氨酸N-Propionylglycine	氨基酸Amino acid	1.33	0.0038	1.30
pme2559	N-乙酰天门冬氨酸N-Acetylaspartate	氨基酸Amino acid	1.27	0.0026	1.28
pme2743	N-苯基乙酰甘氨酸N-Phenylacetylglycine	氨基酸Amino acid	1.39	0.0023	1.31
pmb2591	乙酰色氨酸Acetyltryptophan	氨基酸Amino acid	2.35	0.0000	1.32
pmn001728	N-乙酰-DL-色氨酸N-Acetyl-DL-tryptophan	氨基酸Amino acid	2.15	0.0001	1.32
pma0702	N，N'-双（芥子酰基）亚精胺N，N'-Bis（sinapoyl）Spermidine	多胺Polyamines	11.11	0.0354	1.24
pme2049	2-羟基丁酸2-Hydroxybutanoic acid	有机酸Organic acid	11.09	0.0005	1.32
mws0178	绿原酸Chlorogenic acid	酚酸Phenolic acid	1.06	0.0114	1.24
Rfmb26201	丁香脂醇-乙酰氨基葡萄糖Syringaresinol-aceGlu	糖类Carbohydrate	1.03	0.0308	1.22
Lmjn003562	3，6'-O-二阿魏酰蔗糖3，6'-O-diferuloylsucrose	糖类Carbohydrate	1.15	0.0389	1.15
pmn001399	2，4，6，4'-四羟基二苯乙烯-2-O-D-吡喃葡萄糖苷 2，4，6，4'-Tetrahydroxy-stilbene -2-O-D-glucopyranoside	糖苷Glycoside	1.57	0.0088	1.30
Hmbp003234	1-O-咖啡酰基甘油1-O-Caffeoylglycerol	脂类Lipid	1.01	0.0312	1.20
pmn001318	1，3-O-二-对香豆酰基甘油1，3-O-Di-p-Coumaroyl glycerol	脂类Lipid	3.09	0.0018	1.31
Hmbn005456	1-O-咖啡酰-3-O-对-香豆酰甘油1-O-Caffeoyl-3-O-p-coumaroylglycerol	脂类Lipid	3.74	0.0002	1.32
Cmbn007148	1-O-果糖基-3-O-咖啡酰基甘油1-O-Feruloyl-3-O-caffeoylglycerol	脂类Lipid	4.25	0.0028	1.31
Hmbp006861	1，2-O-二阿魏酰甘油1，2-O-diferuloylglycerol	脂类Lipid	3.72	0.0050	1.30
pmp000091	1，3-二硬脂酸甘油酯Glyceryl 1，3-diferulate	酯类Ester	4.04	0.0275	1.25
mws0853	芥子醇Sinapyl alcohol	苯酚Phenol	2.07	0.0079	1.25
pmn001663	丁香树脂醇Syringaresinol	萜类Terpene	2.24	0.0081	1.28

转录组与代谢组联合分析菠萝网纱覆盖防寒机制

Unveiling the Mechanisms of Pineapple Responding to Anti-chilling by Gauze Covering in Winter via Transcriptome and Metabolome Profiling

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基因名称 Gene name	基因差异倍数 log₂FC	代谢物名称 Metabolite name	代谢物差异倍数 log₂FC	相关系数 Correlation coefficient
水通道蛋白NIP2-2-like^[32]	1.18	丁香脂醇-乙酰氨基葡萄糖 Syringaresinol-aceGlu	1.03	0.83
羟基肉桂酰转移酶 Hydroxycinnamoyltransferase 4-like^[33]	-1.05	绿原酸Chlorogenic acid	1.06	-0.84
羟基肉桂酰转移酶 Hydroxycinnamoyltransferase 4-like^[33]	-1.05	圣草酚Eriodictyol	14.07	-0.96
AP2/ERF^[28]	1.53	1-O-咖啡酰-3-O-对-香豆酰甘油 1-O-Caffeoyl-3-O-p-coumaroylglycerol	3.74	0.87
β-葡萄糖苷酶Beta-glucosidase^[22]	2.50	丁香脂醇-乙酰氨基葡萄糖Syringaresinol-ace Glu	1.03	0.95
	2.50	3，6'-O-二阿魏酰蔗糖3，6'-O-diferuloylsucrose	1.15	0.91
	2.50	1-O-果糖基-3-O-咖啡酰基甘油 1-O-Feruloyl-3-O-caffeoylglycerol	4.25	0.98
硝酸还原酶NADH-like^[23]	3.68	N-苯基乙酰甘氨酸N-Phenylacetylglycine	1.39	0.99
烟草胺转氨酶Nicotianamine aminotransferase A-like^[34]	1.17	N-苯基乙酰甘氨酸N-Phenylacetylglycine	1.39	0.88

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