黄瓜苋科凝集素基因的表达分析与逆境调控

doi:10.11983/CBB20177

植物学报 ›› 2021, Vol. 56 ›› Issue (2): 183-190.DOI: 10.11983/CBB20177

黄瓜苋科凝集素基因的表达分析与逆境调控

赵菲¹, 党刘毅², 魏敏惠¹, 刘春莹¹, 冷伟¹^,^*(), 尚琛晶¹^,³^,^*()

¹陕西中医药大学基础医学院, 咸阳 712046
²西北大学生命科学学院, 西安 710069
³深圳大学生命与海洋科学学院, 深圳 518060

收稿日期:2020-11-01 接受日期:2021-03-01 出版日期:2021-03-01 发布日期:2021-03-17
通讯作者: 冷伟,尚琛晶
作者简介:cjshang@szu.edu.cn
^*E-mail: LWselt@126.com;
基金资助:
陕西省自然科学基础研究计划(2020JQ-866);陕西省教育厅专项科研计划(20JK0590);广东省自然科学基金(2017A030310332);深圳大学新教师科研启动项目(2019078)

Expression of Amaranthin-like Lectins Gene and Responses to Abiotic Stresses in Cucumber

Fei Zhao¹, Liuyi Dang², Minhui Wei¹, Chunying Liu¹, Wei Leng¹^,^*(), Chenjing Shang¹^,³^,^*()

¹College of Basic Medical Sciences, Shaanxi University of Chinese Medicine, Xianyang 712046, China
²College of Life Sciences, Northwest University, Xi’an 710069, China
³College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China

Received:2020-11-01 Accepted:2021-03-01 Online:2021-03-01 Published:2021-03-17
Contact: Wei Leng,Chenjing Shang

摘要/Abstract

摘要： 凝集素是一类具有特异性糖结合活性的蛋白质, 通常具有1个或多个非催化的糖结合结构域。凝集素在植物对病原菌的防御反应中发挥重要作用。由于其抗细菌、真菌、病毒和昆虫等的活性, 凝集素在农业和生物医药领域都具有很大的应用潜力。作为最小的凝集素家族之一, 苋科凝集素的研究较少。该文通过对重要经济作物黄瓜(Cucumis sativus)的基因组进行分析, 对16种苋科凝集素基因在黄瓜基因组中的分布和位置进行研究, 并分析相关基因的外显子/内含子组成。进一步通过启动子分析, 阐明了苋科凝集素基因对非生物胁迫的响应情况。最后, 通过实时荧光定量PCR, 检测了黄瓜中4种苋科凝集素基因对低温、高盐、干旱和ABA处理的响应情况。研究结果可为揭示苋科凝集素的生理功能及其在植物胁迫响应中的作用提供参考。

关键词: 非生物胁迫, 苋科凝集素, 启动子, 蛋白结构域

Abstract: Lectins are globular proteins with carbohydrate-binding sites which enable them to specifically recognize and bind single or more particular carbohydrate structures. Previous studies have shown that lectins play critical roles in plant defense against their herbivores and pathogens. Due to their toxicity towards organisms like bacteria, fungi, viruses and insects, lectins are believed to have great potential in both agricultural and medical applications. As one of the smallest lectin families, there has been very limited research on amaranthin-like lectins. Here, we analyzed the distributions and intron/exon structures from 16 different amaranthin-like genes in the genome of cucumber, an important economic crop. To evaluate their responses against different stresses/plant hormones, promoter analysis was performed for all amaranthin-like genes. Finally, real-time quantitative PCR were performed on stress-treated plants to analyze the responses of amaranthin-like genes towards cold, salt, drought stresses and ABA treatment. This work provides valuable information for the study of physiological roles of amaranthin-like proteins and their involvement in plant defense.

Key words: abiotic stress, amaranthin-like lectins, promoter, protein domain

赵菲, 党刘毅, 魏敏惠, 刘春莹, 冷伟, 尚琛晶. 黄瓜苋科凝集素基因的表达分析与逆境调控. 植物学报, 2021, 56(2): 183-190.

Fei Zhao, Liuyi Dang, Minhui Wei, Chunying Liu, Wei Leng, Chenjing Shang. Expression of Amaranthin-like Lectins Gene and Responses to Abiotic Stresses in Cucumber. Chinese Bulletin of Botany, 2021, 56(2): 183-190.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB20177

https://www.chinbullbotany.com/CN/Y2021/V56/I2/183

图/表 7

表1 实时荧光定量PCR引物列表

Table 1 Primer list for qRT-PCR

Primer name	Forward primer (5'-3')	Reverse primer (5'-3')
CACS	TGGGAAGATTCTTA- TGAAGTGC	CTCGTCAAATTT- ACACATTGGT
PP2A	CAACAGGTGATATT- GGATTATGAT	GCCAGCTCATCC- TCATATAAG
AAT4	TTCGTACACGCAAC- GAGA	TGAAGAGGGTAA- GGCTTG
NAAT1	GAGGAGCTGTGAAA- GGAGCA	CCCTCCACGACA- GTTCCAAT
AAT9	CAGAAACAGCGAAC- CAGAGC	AACTTCATCCCCA- CCGAGTT
AAT14	GGGAATAGAGACGA- TCCGAACT	GCGCAGAAGGCA- GTGTTT

表2 黄瓜中苋科凝集素基因及其在6号染色体上的位置

Table 2 Amaranthin-like genes in cucumber and their locations on chromosome 6

Gene	Start position	End position	Orientation	Gene	Start position	End position	Orientation
CucsaAAT1	5496646	5495195	Reverse	CucsaAAT9	5654159	5656355	Forward
CucsaAAT2	5583941	5582553	Reverse	CucsaAAT10	5681664	5683484	Forward
CucsaAAT3	5590531	5589140	Reverse	CucsaAAT11	5691672	5690257	Reverse
CucsaAAT4	5596447	5595167	Reverse	CucsaAAT12	7077986	7075008	Reverse
CucsaAAT5	5605793	5607184	Forward	CucsaAAT13	7083801	7085207	Forward
CucsaAAT6	5619299	5621364	Forward	CucsaAAT14	7090825	7094963	Forward
CucsaAAT7	5630016	5631894	Forward	CucsaNAAT1	5648115	5646235	Reverse
CucsaAAT8	5636899	5638943	Forward	CucsaNAAT2	5670107	5668263	Reverse

图1 黄瓜中苋科凝集素基因的外显子/内含子组成及分布

Figure 1 Exon/intron composition and distribution of amaranthin-like genes in cucumber

表3 黄瓜中苋科凝集素蛋白的理化性质

Table 3 Physiochemical properties of amaranthin-like proteins in cucumber

Protein	Domain architecture	No. of amino acids	Protein size (kDa)	Theoretical pI
CucsaAAT1	AAT	484	54.9	8.3
CucsaAAT2	AAT	463	53.6	7.9
CucsaAAT3	AAT	463	53.5	8.0
CucsaAAT4	AAT	466	53.7	8.2
CucsaAAT5	AAT	463	53.3	7.5
CucsaAAT6	AAT	464	53.4	7.5
CucsaAAT7	AAT	471	53.5	5.1
CucsaAAT8	AAT	475	54.7	8.3
CucsaAAT9	AAT	474	55.0	6.7
CucsaAAT10	AAT	476	54.9	5.7
CucsaAAT11	AAT	482	55.4	7.0
CucsaAAT12	AAT	502	57.6	9.0
CucsaAAT13	AAT	468	53.9	7.7
CucsaAAT14	AAT	505	58.2	6.8
CucsaNAAT1	NAAT	626	70.4	5.1
CucsaNAAT2	NAAT	614	69.4	4.9

图2 黄瓜中4种苋科凝集素蛋白的结构域构成

Figure 2 Domain architectures of four amaranthin-like proteins in cucumber

图3 苋科凝集素基因中不同启动子应激调节因子的频率

Figure 3 Frequencies of stress regulatory elements in the amaranthin-like genes promoter regions

图4 不同胁迫处理后黄瓜苋科凝集素基因的表达水平 (A)低温处理; (B)高盐处理;(C)干旱处理; (D)脱落酸处理。柱状图代表了2次生物学重复的平均值和标准偏差, 每个重复中包含实验组与对照组各4株植物; 星号表示处理组与对照组之间的统计学显著性(* P<0.05, ** P<0.01, ***P<0.001)。

Figure 4 Relative expression levels for amaranthin-like genes in cucumber plants subjected to different abiotic stress conditions (A)Cold treatment; (B) Salt treatment; (C) Drought treatment;(D) ABA treatment. Bars represent means and standard errors from two independent biological replicates, each replicate containing a pool of 4 plants for stress-treated groups as well as control groups. Asterisks indicate statistically signi?cant differences compared to the plants with mock treatments (* P<0.05,** P<0.01, ***P<0.001).

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黄瓜苋科凝集素基因的表达分析与逆境调控

Expression of Amaranthin-like Lectins Gene and Responses to Abiotic Stresses in Cucumber

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