菠萝不同品种对低温胁迫响应差异的生理代谢机制

doi:10.13560/j.cnki.biotech.bull.1985.2023-0071

生物技术通报 ›› 2023, Vol. 39 ›› Issue (10): 219-230.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0071

菠萝不同品种对低温胁迫响应差异的生理代谢机制

刘传和¹(), 贺涵¹, 何秀古²(), 陈鑫¹, 刘开¹, 邵雪花¹, 赖多¹, 秦健¹, 庄庆礼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:2023-02-01 出版日期:2023-10-26 发布日期:2023-11-28
通讯作者: 何秀古，男，研究员，研究方向：菠萝优质高效种植技术研究与应用推广；E-mail: hexiugu@gdaas.cn
作者简介:刘传和，男，博士，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(2023KJ109);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);广州市科技计划项目(2023E04J0562)

Physiological and Metabolitic Mechanisms of Different Pineapple Cultivars Responding to Low Temperature Stress

LIU Chuan-he¹(), HE Han¹, HE Xiu-gu²(), CHEN Xin¹, LIU Kai¹, SHAO Xue-hua¹, LAI Duo¹, QIN Jian¹, ZHUANG Qing-li¹, 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:2023-02-01 Published:2023-10-26 Online:2023-11-28

摘要/Abstract

摘要：

为探明菠萝不同品种对低温胁迫响应差异的相关机理，本研究以耐寒性强的‘粤甜’和耐寒性弱的‘巴厘’品种为试材，对常温（25℃）及低温（5℃）处理48 h植株的叶片生理指标、基因相对表达量及差异代谢物等进行比较测定。结果表明，低温处理后‘粤甜’的可溶性蛋白、可溶性糖、脯氨酸含量及POD、SOD、CAT酶活性均显著高于‘巴厘’，丙二醛含量显著低于‘巴厘’；POD、CAT基因相对表达量显著高于‘巴厘’，ProDH基因显著低于‘巴厘’；内源生长素、细胞分裂素、茉莉酸、水杨酸及独角金内酯含量高于‘巴厘’，赤霉素、脱落酸含量低于‘巴厘’。代谢组分析表明，低温处理后‘粤甜’与‘巴厘’间共检测出差异代谢物262个，其中黄酮类代谢物最多。KEGG分析表明‘粤甜’‘巴厘’间差异代谢物主要富集在次生代谢物生物合成、类黄酮生物合成、黄酮与黄酮醇生物合成等代谢通路中。筛选出110个低温诱导的‘粤甜’与‘巴厘’差异代谢物，其中41个在常温下差异显著、低温处理后差异倍数增大，69个在常温下无显著差异、低温处理后差异显著；差异代谢物在‘粤甜’中的含量积累均显著高于‘巴厘’。由此可见，渗透调节及抗氧化活性、内源激素及代谢物积累的差异是‘粤甜’与‘巴厘’低温响应差异的重要原因。

关键词: 菠萝, 耐寒性, 不同品种, 植物激素

Abstract:

The aim of this trial is to explore the relevant mechanisms of two different pineapple cultivars responding variously to low temperature stress. Having the pineapple cultivars ‘Yuetian’ with stronger chilling-resistance and ‘Comte de Paris’ with weaker chilling-resistance as materials, the physiological aspects, relative expressions of genes and differentially accumulated metabolites（DAMs）of the pineapple leaves were determined and compared under low temperature（LT, 5℃）and normal temperature（NT, 25℃）treatments for 48 h. The obtained results indicated that the contents of soluble protein, soluble sugar and proline, as well as the activities of POD, SOD and CAT enzymes in ‘Yuetian’ were significantly higher than those in ‘Comte de Paris’ after LT treatment, while the content of MDA was significantly lower than that in ‘Comte de Paris’, respectively. After LT treatment, the relative expressions of POD and CAT genes in ‘Yuetian’ were significantly higher than those in ‘Comte de Paris’, respectively; whereas MDA content was significantly lower than that of ‘Comte de Paris’, and the relative expressions of POD and CAT genes was significantly higher than that of ‘Comte de Paris’. The contents of phytohormones of auxin, cytokinin, jasmonic acid, salicylic acid and strigolactone in ‘Yuetian’ were higher than those in ‘Comte de Paris’, nevertheless the contents of gibberellin and abscisic acid were lower than those in ‘Comte de Paris’ after LT treatment, respectively. Metabolomics analysis showed that a total of 262 DAMs were detected between ‘Yuetian’ and ‘Comte de Paris’ after LT treatment, which predominantly included flavonoid metabolites. KEGG analysis indicated that the DAMs were principally enriched in the three pathways including biosynthesis of secondary metabolites, flavonoid biosynthesis, as well as flavone and flavonol biosynthesis. Further, 110 DAMs were screened between ‘Yuetian’ and ‘Comte de Paris’ e induced by LT treatment, and the accumulating levels of these screened metabolites in ‘Yuetian’ were correspondingly observed to be significantly higher than those in ‘Comte de Paris’. Among the 110 DAMs, 41 had significant differences at NT condition, and the fold changes increased after LT treatment. Besides, 69 had no significant differences at NT condition and had significant differences after LT treatment. Summarily, the diversity in osmotic adjustment and anti-oxidized activity, as well as endogenous hormones and metabolites accumulation might be the vital aspects that result in the various responses to low temperature stress for the two involved pineapple cultivars.

Key words: pineapple（Ananas comosus）, chilling-resistance, different cultivars, phytohormones

刘传和, 贺涵, 何秀古, 陈鑫, 刘开, 邵雪花, 赖多, 秦健, 庄庆礼, 匡石滋, 肖维强. 菠萝不同品种对低温胁迫响应差异的生理代谢机制[J]. 生物技术通报, 2023, 39(10): 219-230.

LIU Chuan-he, HE Han, HE Xiu-gu, CHEN Xin, LIU Kai, SHAO Xue-hua, LAI Duo, QIN Jian, ZHUANG Qing-li, KUANG Shi-zi, XIAO Wei-qiang. Physiological and Metabolitic Mechanisms of Different Pineapple Cultivars Responding to Low Temperature Stress[J]. Biotechnology Bulletin, 2023, 39(10): 219-230.

图/表 8

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基因Gene	ID	正向引物Forward primer（5'-3'）	反向引物Reverse primer（5'-3'）	长度Length/bp
SOD	LOC109709661	CTTCTCTTCCTCCACCTGGTCTCC	TAGGGTTAGGGCTCGCTTGGATG	88
POD	LOC109727354	CTCCGCCTTCACTTCCACGATTG	TGCTGACACGATGAATGACAGACTC	80
CAT	LOC109707674	CTGGCGGATCGTTGACTTCTTCTC	TCCATGTGGCGGTAGTCGGTAG	102
ProDH	LOC109707212	CTTCGCCTATTCACGGGAGCATTC	CAGATCCTTGCCTCACTCGTTCG	90
Actin	LOC109722956	CTGGCCTACGTGGCACTTGACTT	CACTTCTGGGCAGCGGAACCTTT	135

基因Gene	ID	正向引物Forward primer（5'-3'）	反向引物Reverse primer（5'-3'）	长度Length/bp
SOD	LOC109709661	CTTCTCTTCCTCCACCTGGTCTCC	TAGGGTTAGGGCTCGCTTGGATG	88
POD	LOC109727354	CTCCGCCTTCACTTCCACGATTG	TGCTGACACGATGAATGACAGACTC	80
CAT	LOC109707674	CTGGCGGATCGTTGACTTCTTCTC	TCCATGTGGCGGTAGTCGGTAG	102
ProDH	LOC109707212	CTTCGCCTATTCACGGGAGCATTC	CAGATCCTTGCCTCACTCGTTCG	90
Actin	LOC109722956	CTGGCCTACGTGGCACTTGACTT	CACTTCTGGGCAGCGGAACCTTT	135

处理 Treatment	品种 Cultivar	可溶性蛋白 Soluble protein/（mg·kg^-1 FW）	可溶性糖 Soluble sugar/（mg·g^-1 FW）	脯氨酸Proline /（µg·g^-1 FW）	丙二醛MDA /（nmol·g^-1 FW）	过氧化物酶POD /（U·g^-1 FW）	超氧化物酶SOD /（U·g^-1 FW）	过氧化氢酶CAT /（U·g^-1 FW）
常温Normal temperature（NT）	‘粤甜’ ‘Yuetian’	963.00±2.00b	9.78±0.05b	14.10±0.29b	7.94±0.09c	213.99±2.91c	42.23±0.04b	13.93±0.53bc
常温Normal temperature（NT）	‘巴厘’ ‘Comte de Paris’	806.00±3.00c	10.84±0.06a	10.89±0.26d	13.72±0.09b	249.65±7.70b	38.24±0.06d	12.05±1.06c
低温Low temperature（LT）	‘粤甜’ ‘Yuetian’	1056.00±2.00a	9.68±0.12b	23.12±0.31a	7.25±0.08d	356.65±2.92a	43.51±0.06a	24.85±1.84a
低温Low temperature（LT）	‘巴厘’ ‘Comte de Paris’	761.00±8.00d	8.01±0.05c	12.57±0.30c	24.46±0.07a	178.32±2.91d	39.55±0.05c	16.57±1.06b

处理 Treatment	品种 Cultivar	可溶性蛋白 Soluble protein/（mg·kg^-1 FW）	可溶性糖 Soluble sugar/（mg·g^-1 FW）	脯氨酸Proline /（µg·g^-1 FW）	丙二醛MDA /（nmol·g^-1 FW）	过氧化物酶POD /（U·g^-1 FW）	超氧化物酶SOD /（U·g^-1 FW）	过氧化氢酶CAT /（U·g^-1 FW）
常温Normal temperature（NT）	‘粤甜’ ‘Yuetian’	963.00±2.00b	9.78±0.05b	14.10±0.29b	7.94±0.09c	213.99±2.91c	42.23±0.04b	13.93±0.53bc
常温Normal temperature（NT）	‘巴厘’ ‘Comte de Paris’	806.00±3.00c	10.84±0.06a	10.89±0.26d	13.72±0.09b	249.65±7.70b	38.24±0.06d	12.05±1.06c
低温Low temperature（LT）	‘粤甜’ ‘Yuetian’	1056.00±2.00a	9.68±0.12b	23.12±0.31a	7.25±0.08d	356.65±2.92a	43.51±0.06a	24.85±1.84a
低温Low temperature（LT）	‘巴厘’ ‘Comte de Paris’	761.00±8.00d	8.01±0.05c	12.57±0.30c	24.46±0.07a	178.32±2.91d	39.55±0.05c	16.57±1.06b

分类Class	物质Compound	常温NT		低温LT
分类Class	物质Compound	‘粤甜’‘Yuetian’	‘巴厘’‘Comte de Paris’	‘粤甜’‘Yuetian’	‘巴厘’‘Comte de Paris’
生长素 Auxin	色胺Tryptamine	9.77±1.19b	2.59±0.65c	16.78±0.51a	2.06±0.97c
	L-色氨酸L-tryptophan	1422.30±186.94ab	1072.91±83.88b	1734.66±127.79a	946.77±157.24b
	吲哚-3-乙酸甲酯 Methyl indole-3-acetate	0.27±0.02ab	0.19±0.01bc	0.35±0.05a	0.14±0.03c
	吲哚-3-甲醛Indole-3-carboxaldehyde	4.17±0.10a	4.64±0.37a	4.87±0.40a	3.53±0.61a
	吲哚-3-甲酸Indole-3-carboxylic acid	0.73±0.07b	1.39±0.11a	1.19±0.15a	0.76±0.07b
	吲哚乙酸-缬氨酸甲酯 Indole-3-acetyl-L-valine methyl ester	0.031±0.004a	0.026±0.002a	0.038±0.010a	0.016±0.002a
	吲哚乙酸-天冬氨酸 Indole-3-acetyl-L-aspartic acid	9.32±1.92b	2.31±0.4b	21.69±3.69a	3.49±2.2b
赤霉素GA	赤霉素19 GA₁₉	3.19±0.24b	6.19±0.28a	2.87±0.23b	5.67±0.48a
细胞分裂素 Cytokinin	2-甲硫基顺式玉米素核苷 2-Methylthio-cis-zeatin riboside	0.19±0ab	0.17±0.01b	0.23±0.02a	0.16±0.01b
	异戊烯腺嘌呤核苷 N6-isopentenyladenosine	1.12±0.07b	0.45±0.07c	2.00±0.21a	0.25±0.05c
	双氢玉米素-7-糖苷 Dihydrozeatin-7-glucoside	0.32±0.06a	0.16±0.02a	0.34±0.02a	0.18±0.05a
	6-苄氨基嘌呤6-Benzyladenine	0.13±0.02a	0.11±0.02a	0.15±0.02a	0.11±0.01a
乙烯类 Ethylene	1-氨基环丙烷羧酸 1-Aminocyclopropanecarboxylic acid（ACC）	42.80±2.07a	45.03±1.48a	42.75±0.92a	34.00±1.04b
脱落酸 Abscisic acid	脱落酸Abscisic acid	10.34±0.27bc	29.64±0.54a	7.92±0.26c	14.29±2.03b
脱落酸 Abscisic acid	脱落酸葡萄糖酯 ABA-glucosyl ester	118.57±6.49a	189.16±41.33a	119.57±4.59a	174.47±2.04a
茉莉酸 Jasmonic acid	二氢茉莉酸Dihydrojasmonic acid	0.09±0.04a	0.09±0.02a	0.10±0.01a	0.04±0.01a
	茉莉酸-异亮氨酸 Jasmonoyl-L-isoleucine	0.63±0.09a	0.36±0.19ab	0.23±0.07ab	0.08±0.03b
	茉莉酸Jasmonic acid	2.61±0.34a	1.48±0.48ab	0.43±0.06b	0.40±0.03b
	顺式-12-氧-植物-二烯酸 Cis（+）-12-Oxophytodienoic acid	2.95±0.25a	2.27±0.18a	1.24±0.11b	0.93±0.12b
水杨酸 Salicylic acid	水杨酸Salicylic acid	35.42±1.64a	22.87±0.95b	28.39±2.89ab	14.59±0.98c
水杨酸 Salicylic acid	水杨酸-2-O-β-葡萄糖苷 Salicylic acid 2-O-β-glucoside	1135.25±119.94a	219.08±7.58b	1250.95±62.61a	211.29±25.88b
独角金内酯 Strigolactone	5-脱氧独脚金醇5-Deoxystrigol	73.07±4.24a	68.29±3.38ab	66.97±2.03ab	54.83±3.91b

菠萝不同品种对低温胁迫响应差异的生理代谢机制

Physiological and Metabolitic Mechanisms of Different Pineapple Cultivars Responding to Low Temperature Stress

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