菠萝Dof转录因子在己醛延缓果实黑心病发生中表达分析

doi:10.11924/j.issn.1000-6850.casb20191100794

中国农学通报 ›› 2020, Vol. 36 ›› Issue (15): 115-122.doi: 10.11924/j.issn.1000-6850.casb20191100794

所属专题：生物技术

菠萝Dof转录因子在己醛延缓果实黑心病发生中表达分析

陈丽, 谷会, 贾志伟, 洪克前()

中国热带农业科学院南亚热带作物研究所,海南省热带园艺产品采后生理与保鲜重点实验室,广东湛江 524091

收稿日期:2019-11-07 修回日期:2019-11-28 出版日期:2020-05-25 发布日期:2020-05-21
通讯作者: 洪克前
作者简介:陈丽,女,1971年出生,广东湛江人,研究实习员,硕士,研究方向：食品科学。通信地址：524091 广东省湛江市麻章区湖秀路一号（湖光岩北）南亚热带作物研究所,Tel：0759-2858198,E-mail：chenroumu@126.com。
基金资助:
中央级公益性科研院所基本科研业务费专项一中国热带农业科学院院级创新团队项目“保鲜新技术开发”(N0.1630062017032);“菠萝采后病害防控及保鲜技术研究”(N0.1630062017031)

Dof Transcription Factor in Pineapple Fruits Treated with Hexaldehyde Delaying Internal Browning Resistance: Expression Analysis

Chen Li, Gu Hui, Jia Zhiwei, Hong Keqian()

South Subtropical Crops Research Institutes, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory for Postharvest Physiology and Technology of Tropical Horticultural Products of Hainan Province, Zhanjiang Guangdong 524091

Received:2019-11-07 Revised:2019-11-28 Online:2020-05-25 Published:2020-05-21
Contact: Hong Keqian

摘要/Abstract

摘要：

Dof蛋白是植物所特有的一类转录因子,在许多生物及非生物胁迫反应中扮演着重要的角色。本研究旨在探讨菠萝AcDof转录因子家族成员在果实诱导抗黑心病中的作用。以菠萝果实为试材,采用延缓菠萝黑心病发生的外源己醛处理果实,处理结束后放置在25℃下贮藏8天,通过荧光定量PCR(qRT-PCR)技术分析菠萝AcDofs转录因子在果实抗黑心病诱导中的基因表达变化。基于菠萝基因组数据库,筛选并克隆到25个Dof转录因子,被命名为AcDof1-AcDof25;0.1%(v/v)己醛处理显著延缓了菠萝果实黑心指数的上升;qRT-PCR分析表明,20个AcDof转录因子家族成员中,有18个AcDof转录因子被己醛处理所激活,其中AcDof4、AcDof8、AcDof14、AcDof16和AcDof18等5个转录因子在处理组和对照组果实中表达趋势有较大差异;AcDof13被己醛处理显著抑制。推测这6个AcDof转录因子以不同方式响应了己醛延缓菠萝果实黑心病发生过程,它们可能是黑心病发生的关联基因。

关键词: Dof, 基因表达, 己醛, 黑心病, 菠萝果实

Abstract:

DNA binding with one finger (Dof) proteins, which are a family of plant-specific transcription factor (TF), plays important roles in diverse biotic and abiotic stress responses. The purpose of this study is to investigate the roles of Dof TF family members in internal browning (IB) resistance in the postharvest pineapple fruits. Pineapple fruits were used as materials, the pineapple fruits with IB were treated with the exogenous hexaldehyde, and then stored the treated at 25 ℃ for 8 days. Quantitative real-time PCR (qRT-PCR) technique was adopted to analyze the changes in Dof TFs gene expressions during the storage duration of fruits inducted by IB resistance. 25 putative Dof TFs which were designated as AcDof1-AcDof25, were identified and isolated based on pineapple genome database. 0.1% (v/v) hexaldehyde treatment significantly delayed the ascent of IB index of fruits compared with the controls. qRT-PCR analysis showed that 18 out of 20 AcDof TFs were up-regulated after hexaldehyde treatment. Among them, 5 TFs, such as AcDof4, AcDof8, AcDof14, AcDof16 and AcDof18 had significant expression profiles between controls and the treated fruits. However, 1 AcDof, namely AcDof13, was apparently down-regulated by hexaldehyde treatment. These 6 AcDofs might be involved in the regulation of hexaldehyde delaying IB in different ways, thus they could be the related genes of IB.

Key words: Dof, gene expression, hexaldehyde, internal browning, pineapple fruit

中图分类号:

Q786

陈丽, 谷会, 贾志伟, 洪克前. 菠萝Dof转录因子在己醛延缓果实黑心病发生中表达分析[J]. 中国农学通报, 2020, 36(15): 115-122.

Chen Li, Gu Hui, Jia Zhiwei, Hong Keqian. Dof Transcription Factor in Pineapple Fruits Treated with Hexaldehyde Delaying Internal Browning Resistance: Expression Analysis[J]. Chinese Agricultural Science Bulletin, 2020, 36(15): 115-122.

图/表 5

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基因	正向引物(5′-3′)	反向引物(5′-3′)
Acactin	CTGGCCTACGTGGCACTTGACTT	CACTTCTGGGCAGCGGAACCTTT
AcDof1	ACCATTGTACCCTGCACCTTAC	TTGATCCCATTTCCATCCCT
AcDof2	CAGAAGCAGAAGCAGAAGCAG	CAATGGGAAGCGATAATGGC
AcDof3	CTTGGCTTTCCCCTACCACA	GGGCACAAACGACCCTACTC
AcDof4	GCAAGAGAGGAAGGTGGTGAG	TGGAAGAGGAGTTGGATGAGG
AcDof8	AAGAATGAGAAGGTGATGGA	GGTCGGATTTAGGGCAA
AcDof10	CAACCTCTCCTTCCCACA	TCATTTCCATCACCTCCTC
AcDof11	TACGGCGAAAATGGAAGTGT	CCCCCAGAAGAAGGAAAGAAAT
AcDof12	CCGACAAAGAGCGAAGGGA	GCAGACCAAAGCAGGAGACC
AcDof13	CCGGGAGACGAAGTTCT	CGTTGTTGATGATGATGATG
AcDof14	ATCTCTGAGGAATGTCCCTGTTG	GGTGATGCTGAGGTTGGTGG
AcDof15	ACGCAACAAGCGGAGCA	TGTGGAGGAAGAGGAGGACG
AcDof16	CATCCACCATCCCTTTCCC	AACTCTTTGCCGCCTCATCT
AcDof17	ATGCCGCTCAAACTGC	CCTTCCCTCTCTCTCCTCT
AcDof18	AACCAGGGAGTAGCG	GTAGTTGTTGTAGTAGCAGA
AcDof19	ATAAGGAGGAATCAATGGAG	TTGGCGGGTAATAAGGTA
AcDof20	CTTGGGAAGGAGGCGTTTT	GAGGGTTTTTGCGGAGTTTT
AcDof21	TTGACCCCCTCATTCCACC	CTCTGTTTTTCGTTTTCCGCTT
AcDof22	TGGCTTCCCCAACCTCTACT	CCTCCTTCCAACATTGCTTTAT
AcDof23	TGCCGACACCACTGAAGACA	AAGACGACACGGAGGAGAAAG
AcDof25	CAAATCACAAGCAGCCTC	AACTATCAATCAACCACGC

基因	正向引物(5′-3′)	反向引物(5′-3′)
Acactin	CTGGCCTACGTGGCACTTGACTT	CACTTCTGGGCAGCGGAACCTTT
AcDof1	ACCATTGTACCCTGCACCTTAC	TTGATCCCATTTCCATCCCT
AcDof2	CAGAAGCAGAAGCAGAAGCAG	CAATGGGAAGCGATAATGGC
AcDof3	CTTGGCTTTCCCCTACCACA	GGGCACAAACGACCCTACTC
AcDof4	GCAAGAGAGGAAGGTGGTGAG	TGGAAGAGGAGTTGGATGAGG
AcDof8	AAGAATGAGAAGGTGATGGA	GGTCGGATTTAGGGCAA
AcDof10	CAACCTCTCCTTCCCACA	TCATTTCCATCACCTCCTC
AcDof11	TACGGCGAAAATGGAAGTGT	CCCCCAGAAGAAGGAAAGAAAT
AcDof12	CCGACAAAGAGCGAAGGGA	GCAGACCAAAGCAGGAGACC
AcDof13	CCGGGAGACGAAGTTCT	CGTTGTTGATGATGATGATG
AcDof14	ATCTCTGAGGAATGTCCCTGTTG	GGTGATGCTGAGGTTGGTGG
AcDof15	ACGCAACAAGCGGAGCA	TGTGGAGGAAGAGGAGGACG
AcDof16	CATCCACCATCCCTTTCCC	AACTCTTTGCCGCCTCATCT
AcDof17	ATGCCGCTCAAACTGC	CCTTCCCTCTCTCTCCTCT
AcDof18	AACCAGGGAGTAGCG	GTAGTTGTTGTAGTAGCAGA
AcDof19	ATAAGGAGGAATCAATGGAG	TTGGCGGGTAATAAGGTA
AcDof20	CTTGGGAAGGAGGCGTTTT	GAGGGTTTTTGCGGAGTTTT
AcDof21	TTGACCCCCTCATTCCACC	CTCTGTTTTTCGTTTTCCGCTT
AcDof22	TGGCTTCCCCAACCTCTACT	CCTCCTTCCAACATTGCTTTAT
AcDof23	TGCCGACACCACTGAAGACA	AAGACGACACGGAGGAGAAAG
AcDof25	CAAATCACAAGCAGCCTC	AACTATCAATCAACCACGC

基因	氨基酸(AA)	起始	终止	E值
AcDof1	428	114	172	2.5e-33
AcDof2	259	31	89	7.4e-33
AcDof3	408	52	110	2.8e-33
AcDof4	364	31	89	2.5e-33
AcDof5	272	72	130	9.4e-36
AcDof6	361	63	121	1.5e-35
AcDof7	180	22	83	7.3e-30
AcDof8	354	92	150	3.9e-36
AcDof9	285	45	102	9.2e-34
AcDof10	336	53	111	4.4e-36
AcDof11	259	34	92	3.6e-36
AcDof12	361	148	206	3.7e-30
AcDof13	213	85	138	4.6e-18
AcDof14	307	50	108	2.6e-36
AcDof15	363	83	141	1.3e-35
AcDof16	497	138	196	3.8e-35
AcDof17	221	117	174	3e-17
AcDof18	136	79	135	4.2e-32
AcDof19	478	131	189	7.1e-34
AcDof20	146	77	109	8e-12
AcDof21	381	97	155	1.4e-34
AcDof22	362	53	111	1.4e-33
AcDof23	212	27	85	9.3e-33
AcDof24	326	107	152	4.2e-19
AcDof25	368	35	93	7.6e-36

基因	氨基酸(AA)	起始	终止	E值
AcDof1	428	114	172	2.5e-33
AcDof2	259	31	89	7.4e-33
AcDof3	408	52	110	2.8e-33
AcDof4	364	31	89	2.5e-33
AcDof5	272	72	130	9.4e-36
AcDof6	361	63	121	1.5e-35
AcDof7	180	22	83	7.3e-30
AcDof8	354	92	150	3.9e-36
AcDof9	285	45	102	9.2e-34
AcDof10	336	53	111	4.4e-36
AcDof11	259	34	92	3.6e-36
AcDof12	361	148	206	3.7e-30
AcDof13	213	85	138	4.6e-18
AcDof14	307	50	108	2.6e-36
AcDof15	363	83	141	1.3e-35
AcDof16	497	138	196	3.8e-35
AcDof17	221	117	174	3e-17
AcDof18	136	79	135	4.2e-32
AcDof19	478	131	189	7.1e-34
AcDof20	146	77	109	8e-12
AcDof21	381	97	155	1.4e-34
AcDof22	362	53	111	1.4e-33
AcDof23	212	27	85	9.3e-33
AcDof24	326	107	152	4.2e-19
AcDof25	368	35	93	7.6e-36

基因		处理		贮藏时间
基因		处理		0 d		2 d	4 d	6 d	8 d
Dof1		对照		1.00 ± 0.024		0.54 ± 0.04^*	0.38 ± 0.045^*	0.17 ± 0.014^*	0.15 ± 0.015^*
Dof1		己醛		1.00 ± 0.084		0.74 ± 0.048	0.45 ± 0.055	0.29± 0.024	0.33 ± 0.015
Dof 2		对照		1.00 ± 0.054		0.91 ± 0.04	0.10 ± 0.045^*	0.17 ± 0.014^*	0.31 ± 0.015
Dof 2		己醛		1.00 ± 0.044		0.66 ± 0.048^*	0.89 ± 0.055	0.36 ± 0.024	0.32 ± 0.015
Dof 3		对照		1.00 ± 0.06		0.24 ± 0.04^*	0.48 ± 0.05^*	0.37 ± 0.033^*	0.23 ± 0.018
Dof 3		己醛		1.00 ± 0.08		0.58 ± 0.06	0.74 ± 0.035	1.03 ± 0.078	0.038 ± 0.0095^*
Dof 4		对照		1.00 ± 0.049		0.11 ± 0.084^*	0.07 ± 0.021^*	0.07 ± 0.0038^*	0.23 ± 0.048
Dof 4		己醛		1.00 ± 0.07		0.45 ± 0.028	1.89 ± 0. 49	4.67 ± 0.21	0.027 ± 0.012^*
Dof 8		对照		1.00 ± 0.027		0.57 ± 0.042^*	0.66 ± 0.01^*	0.85 ± 0.09^*	0.57 ± 0.07
Dof 8		己醛		1.00 ± 0.031		1.10 ± 0. 37	0.98 ± 0.09	1.71 ± 0.31^*	0.13 ± 0.01^*
Dof 10		对照		1.00 ± 0.14		0.94 ± 0.039	0.90 ± 0.089	0.57 ± 0.08^*	1.31 ± 0.15^*
Dof 10		己醛		1.00 ± 0.13		0.92 ± 0.046	0.94 ± 0.05	2.18 ± 0.25	1.74 ± 0.26
Dof 11		对照		1.00 ± 0.078		1.11 ± 0.12^*	1.61 ± 0.34	0.92 ± 0.28^*	0.76 ± 0.15^*
Dof 11		己醛		1.00 ± 0.089		1.68 ± 0.18	1.74 ± 0.15	1.36 ± 0.17	1.84 ± 0.14
Dof 12		对照		1.00 ± 0.044		1.19 ± 0.086	1.51 ± 0.15	0.57 ± 0.036	0.40 ± 0.025
Dof 12		己醛		1.00 ± 0.037		1.13 ± 0.076	1.38 ± 0.13	0.63 ± 0.028	0.44 ± 0.039
Dof 13		对照		1.00 ± 0.29		0.61 ± 0.049	0.54 ± 0.014	2.16 ± 0.48	2.29 ± 0.18
Dof 13		己醛		1.00 ± 0.034		0.29 ± 0.024^*	0.28 ± 0.042^*	1.39 ± 0.21^*	0.53 ± 0.077^*
Dof 14		对照		1.00 ± 0.035		1.00 ± 0.061^*	0.73 ± 0.14^*	0.44 ± 0.025^*	0.25 ± 0.069^*
Dof 14		己醛		1.00 ± 0.48		1.21 ± 0.14	1.03 ± 0.038	0.68 ± 0.087	0.90 ± 0.041
Dof 15		对照		1.00 ± 0.045		1.00 ± 0.21^*	1.05 ± 0.018	0.93 ± 0.014^*	0.44 ± 0.015^*
Dof 15		己醛		1.00 ± 0.051		1.33 ± 0.11	1.09 ± 0.087	1.61 ± 0.13	0.61 ± 0.015
Dof 16		对照		1.00 ± 0.038		1.59 ± 0.013^*	0.42 ± 0.012^*	0.26 ± 0.014^*	0.60 ± 0.015^*
Dof 16		己醛		1.00 ± 0.044		2.42 ± 0.011	1.33 ± 0.021	0.85 ± 0.011	0.80 ± 0.013
Dof 17		对照		1.00 ± 0.071		0.94 ± 0.045^*	0.61 ± 0.021^*	0.53 ± 0.014^*	0.58 ± 0.021^*
Dof 17		己醛		1.00 ± 0.074		1.02 ± 0.018	0.82 ± 0.014	0.88 ± 0.017	0.67 ± 0.011
Dof 18		对照		1.00 ± 0.15		0.95 ± 0.014^*	1.13 ± 0.017^*	1.59 ± 0.014^*	1.30 ± 0.022
Dof 18		己醛		1.00 ± 0.23		4.12 ± 0.047	2.08 ± 0.028	4.91 ± 0.016	0.11 ± 0.011^*
Dof 19	对照		1.00 ± 0.075		1.77 ± 0.044		2.24 ± 0.024^*	1.33 ± 0.054^*	1.27 ± 0.018
Dof 19	己醛		1.00 ± 0.025		0.54 ± 0.0014^*		2.90 ± 0.047	2.07 ± 0.061	1.14 ± 0.024^*
Dof 20	对照		1.00 ± 0.035		0.81 ± 0.014		0.90 ± 0.021^*	0.47 ± 0.011^*	0.62 ± 0.024
Dof 20	己醛		1.00 ± 0.026		0.86 ± 0.013		2.43 ± 0.34	1.69 ± 0.14	0.51 ± 0.021^*
Dof 21	对照		1.00 ± 0.14		1.54 ± 0.017		1.11 ± 0.008^*	0.36 ± 0.014^*	0.18 ± 0.032^*
Dof 21	己醛		1.00 ± 0.015		1.48 ± 0.047		1.41 ± 0.009	0.73 ± 0.031	0.86 ± 0.025
Dof 22	对照		1.00 ± 0.023		0.49 ± 0.021		0.45 ± 0.014^*	0.23 ± 0.022^*	0.31 ± 0.025^*
Dof 22	己醛		1.00 ± 0.024		0.56 ± 0.033		0.64 ± 0.049	0.59 ± 0.078	0.45 ± 0.034
Dof 23	对照		1.00 ± 0.035		0.51 ± 0.014		0.64 ± 0.021^*	0.31 ± 0.017^*	0.20 ± 0.014
Dof 23	己醛		1.00 ± 0.25		0.54 ± 0.074		0.76 ± 0.014	0.70 ± 0.045	0.053 ± 0.0017^*
Dof 25	对照		1.00 ± 0.085		3.06 ± 0.24		1.38 ± 0.014^*	0.19 ± 0.0016^*	0.57 ± 0.016
Dof 25	己醛		1.00 ± 0.075		2.04 ± 0.17^*		1.79 ± 0.14	1.23 ± 0.017	0.31 ± 0.0024^*

菠萝Dof转录因子在己醛延缓果实黑心病发生中表达分析

Dof Transcription Factor in Pineapple Fruits Treated with Hexaldehyde Delaying Internal Browning Resistance: Expression Analysis

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