JAZ家族基因在采后黄瓜低温贮藏条件下的表达分析

doi:10.3969/j.issn.1004-1524.20231044

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (8): 1820-1831.DOI: 10.3969/j.issn.1004-1524.20231044

JAZ家族基因在采后黄瓜低温贮藏条件下的表达分析

袁晓¹^,²(), 蒋园园¹, 朱云娜¹, 曲姗姗³^,^*(), 王玉昆¹, 原远¹, 王斌¹^,^*()

1.广东省粤北食药资源利用与保护重点实验室/韶关学院生物与农业学院,广东韶关 512005
2.华南农业大学园艺学院/广东省果蔬保鲜重点实验室,广东广州 510642
3.河南农业大学食品科学技术学院,河南郑州 450002

收稿日期:2023-08-31 出版日期:2024-08-25 发布日期:2024-09-06
作者简介:王斌,E-mail: b_wang@sgu.edu.cn
*曲姗姗,E-mail: ssqu1994@henau.edu.cn;
袁晓(1989—),女,广东湛江人,硕士研究生,主要研究方向为采后果蔬贮藏保鲜。E-mail: yxiao@sgu.edu.cn
通讯作者: 曲姗姗,王斌
基金资助:
广东省基础与应用基础研究基金(2022A1515011913)

Expression analysis of JAZ family genes in harvested cucumber fruit under cold storage condition

YUAN Xiao¹^,²(), JIANG Yuanyuan¹, ZHU Yunna¹, QU Shanshan³^,^*(), WANG Yukun¹, YUAN Yuan¹, WANG Bin¹^,^*()

1. Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region/College of Biology and Agriculture, Shaoguan University, Shaoguan 512005, Guangdong, China
2. College of Horticulture, South China Agricultural University/Guangdong Province Key Laboratory of Postharvest Science of Fruits and Vegetables, Guangzhou 510642, China
3. College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China

Received:2023-08-31 Online:2024-08-25 Published:2024-09-06
Contact: QU Shanshan,WANG Bin

摘要/Abstract

摘要：

冷害是限制采后黄瓜低温贮运的主要问题,茉莉酸(jasmonic acid, JA)在植物抗冷性调控中具有重要作用,JAZ(JA ZIM-domain)蛋白是JA信号的转录阻遏物,抑制JA合成和信号传递。为探究JAZ家族基因在采后黄瓜低温贮藏期间的作用,从黄瓜基因组中筛选鉴定了JAZ家族基因,并分析了该家族基因的序列特征、理化性质和时空表达模式。结果显示,黄瓜基因组中共鉴定到了17个JAZ基因,可分为8个亚家族,蛋白序列中均含有Tify或CCT保守结构域。绝大多数的JAZ基因响应贮藏低温(5 ℃和10 ℃)处理,说明JAZ家族蛋白参与采后黄瓜耐冷性调控,但不同JAZ基因对低温处理的响应强度有差异。黄瓜JAZ家族基因的启动子含有丰富的逆境响应元件,但具体的元件类型存在差异。14个JAZ基因的表达量在5 ℃贮藏期间逐渐下降,但JAZ11表达随贮藏时间显著增加。外源脱落酸(abscisic acid, ABA)、茉莉酸甲酯(methyl jasmonate, MeJA)和褪黑素(melatonin, MT)处理,以及采后黄瓜中沉默HHO2和GRP3表达显著影响冷藏黄瓜JAZ家族基因的表达,再次证明黄瓜JAZ家族基因参与采后黄瓜耐冷性的调控。ABA、MeJA和MT处理显著下调JAZ11表达,沉默HHO2和GRP3显著上调JAZ11表达,证明JAZ11负调控JA信号和采后黄瓜耐冷性。该研究明确了黄瓜JAZ家族基因的理化性质和在低温下的表达模式,表明JAZ11可能是JAZ家族蛋白中负调控采后黄瓜耐冷性的主效因子。

关键词: JAZ基因, 采后黄瓜, 低温贮藏, 表达分析

Abstract:

Chilling injury poses a significant challenge to the postharvest storage and transportation of cucumber fruits. Jasmonic acid (JA) plays a crucial role in regulating the chilling resistance in plants. JAZ (JA ZIM-domain) proteins act as transcription repressors of JA signaling, inhibiting both the synthesis and transmission of JA. To explore the role of JAZ family genes during cold storage of harvested cucumber fruits, JAZ family genes were identified and screened from the cucumber genome, and their sequence features, physicochemical properties, and spatial-temporal expression patterns were analyzed. The findings revealed a total of 17 JAZ genes in the cucumber genome, which could be divided into 8 subfamilies, all containing the conserved Tify or CCT domains in their protein sequences. Most of these JAZ genes responded to cold storage (5 ℃ and 10 ℃) treatments, suggesting the involvement of JAZ family proteins in regulating the chilling tolerance in cucumber fruits. However, different JAZ genes exhibited varying degrees of response to cold treatments. The promoters of cucumber JAZ family genes had numerous stress-responsive elements, though the specific types of elements varied. During 5 ℃ storage period, the expression levels of 14 JAZ genes gradually decreased, while the expression of JAZ11 significantly increased with storage time. Exogenous treatments with abscisic acid (ABA), methyl jasmonate (MeJA), melatonin (MT), as well as the silencing HHO2 and GRP3 expression in harvested cucumbers, significantly impacted the expression of JAZ family genes. These results further confirmed the involvement of cucumber JAZ family genes in regulating chilling tolerance. ABA, MeJA, and MT treatments significantly downregulated JAZ11 expression, whereas silencing HHO2 and GRP3 significantly upregulated its expression. These results together suggest that JAZ11 negatively regulates JA signaling and cold tolerance in cucumber fruits. This study provides novel insights into the physicochemical properties and expression patterns of cucumber JAZ family genes under cold conditions, and suggesting that JAZ11 may be the key factor responsible for negative regulation of chilling tolerance in harvested cucumbers among JAZ family proteins.

Key words: JAZ gene, harvested cucumber, cold storage, expression analysis

中图分类号:

S642.2

袁晓, 蒋园园, 朱云娜, 曲姗姗, 王玉昆, 原远, 王斌. JAZ家族基因在采后黄瓜低温贮藏条件下的表达分析[J]. 浙江农业学报, 2024, 36(8): 1820-1831.

YUAN Xiao, JIANG Yuanyuan, ZHU Yunna, QU Shanshan, WANG Yukun, YUAN Yuan, WANG Bin. Expression analysis of JAZ family genes in harvested cucumber fruit under cold storage condition[J]. Acta Agriculturae Zhejiangensis, 2024, 36(8): 1820-1831.

图/表 8

表1 黄瓜JAZ家族基因登录号与基本特性

Table 1 Accession number and basic characteristics of cucumber JAZ family genes

基因名称 Gene name	登录号 Accession number	氨基酸 Amino acid
基因名称 Gene name	登录号 Accession number	序列长度 Sequence length	分子量 Molecular weight/ku	等电点 Isoelectric point	不稳定系数 Instability index	脂肪指数 Aliphatic index	亲水性均值 Grand average of hydropathicity	亚细胞定位 Subcellular localization
JAZ1	CsaV3_3G030830	209	22.87	8.94	52.09	62.2	-0.63	细胞核Nucleus
JAZ2	CsaV3_2G030170	352	38.58	4.86	44.39	62.84	-0.71	细胞核Nucleus
JAZ3	CsaV3_4G002460	200	22.49	6.98	62.80	74.60	-0.55	细胞核Nucleus
JAZ4	CsaV3_5G037080	381	39.67	9.30	44.47	68.22	-0.29	细胞核Nucleus
JAZ5	CsaV3_7G033740	303	33.95	6.39	44.74	64.65	-0.87	细胞核Nucleus
JAZ6	CsaV3_7G034270	295	32.09	9.31	52.17	60.98	-0.76	细胞核Nucleus
JAZ7	CsaV3_4G007460	129	14.86	10.42	57.84	68.84	-0.43	细胞核Nucleus
JAZ8	CsaV3_1G032370	119	13.28	10.38	78.91	80.42	-0.44	细胞核Nucleus
JAZ9	CsaV3_2G030180	279	30.47	8.69	45.54	73.01	-0.51	细胞核Nucleus
JAZ10	CsaV3_6G051810	132	14.74	9.50	74.04	64.32	-0.81	细胞核Nucleus
JAZ11	CsaV3_6G007840	184	20.31	9.41	43.64	88.1	-0.32	细胞核Nucleus
JAZ12	CsaV3_1G041270	231	25.03	9.98	71.00	67.97	-0.46	细胞核Nucleus
JAZ13	CsaV3_7G006810	313	33.67	5.13	42.90	69.14	-0.62	细胞核Nucleus
JAZ14	CsaV3_2G013410	336	36.97	6.71	53.52	65.6	-0.75	细胞核Nucleus
JAZ15	CsaV3_1G007260	339	36.49	9.10	53.87	72.51	-0.24	细胞核Nucleus
JAZ16	CsaV3_2G031660	274	28.70	9.30	44.57	66.28	-0.51	细胞核Nucleus
JAZ17	CsaV3_3G046630	376	40.25	8.64	43.82	62.61	-0.62	细胞核Nucleus

图1 黄瓜JAZ家族蛋白氨基酸序列motif(A)和保守域(B)鉴定

Fig.1 Identification of motif (A) and conserved domain (B) in the amino acid sequence of cucumber JAZ family proteins

图2 黄瓜和拟南芥JAZ家族蛋白系统发育分析

Fig.2 Phylogenetic analysis of JAZ family proteins in cucumber and Arabidopsis

图3 不同低温处理对黄瓜果实JAZ家族基因表达的影响

Fig.3 Effects of different cold treatments on the expression of JAZ family genes in cucumber fruits

图4 黄瓜JAZ家族基因启动子中的顺式作用元件

Fig.4 cis-elements in the promoter of cucumber JAZ family genes

图5 采后黄瓜5 ℃贮藏期间JAZ家族基因表达模式

Fig.5 Expression patterns of JAZ family gene during storage at 5 ℃ in postharvest cucumber

图6 ABA、MeJA和MT处理对低温贮藏条件下黄瓜JAZ家族基因表达的影响

Fig.6 Effects of ABA, MeJA, and MT treatments on the expression of JAZ family genes in cucumber fruits under low temperature storage conditions

图7 采后黄瓜中沉默HHO2和GRP3对低温条件下JAZ家族基因表达的影响 CK,过表达TRV1载体;V-GRP3,沉默GRP3;V-HHO2,沉默HHO2;V-G+H,同时沉默GRP3和HHO2基因。

Fig.7 Effects of silencing HHO2 and GRP3 on the expression of JAZ family genes under low temperature in postharvest cucumber CK: Overexpression of TRV1 vector; V-GRP3: Silencing GRP3; V-HHO2: Silencing HHO2; V-G+H: Simultaneous silencing of GRP3 and HHO2 genes.

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JAZ家族基因在采后黄瓜低温贮藏条件下的表达分析

Expression analysis of JAZ family genes in harvested cucumber fruit under cold storage condition

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