基于RT-qPCR技术解析齿肋赤藓耐热基因在不同非生物胁迫下的表达模式

doi:10.13866/j.azr.2025.05.11

干旱区研究 ›› 2025, Vol. 42 ›› Issue (5): 885-894.doi: 10.13866/j.azr.2025.05.11 cstr: 32277.14.AZR.20250511

基于RT-qPCR技术解析齿肋赤藓耐热基因在不同非生物胁迫下的表达模式

霍雯婷¹(), 顾天麒¹, 高梦宇¹, 宋艳芳², 李鸿彬¹, 卓露¹()

1.石河子大学生命科学学院，新疆植物药资源利用教育部重点实验室，绿洲城镇与山盆系统生态兵团重点实验室，新疆石河子 832000
2.石河子卫生学校，新疆石河子 832000

收稿日期:2025-01-10 修回日期:2025-03-26 出版日期:2025-05-15 发布日期:2025-10-22
通讯作者: 卓露. E-mail: Luzhuo@shzu.edu.cn
作者简介:霍雯婷（1997-），女，硕士研究生，主要从事荒漠藓类植物抗逆生理及分子生物学研究. E-mail: huowenting@stu.shzu.edu.cn
基金资助:
国家自然科学基金项目(32360097);2023年兵团发展专项-博士后专项项目(CZ00150102)

Analysis of the expression patterns of the heat-resistant gene of Syntrichia caninervis under different abiotic stresses based on RT-qPCR technology

HUO Wenting¹(), GU Tianqi¹, GAO Mengyu¹, SONG Yanfang², LI Hongbin¹, ZHUO Lu¹()

1. College of Life Science, Shihezi University, Ministry of Education Key Laboratory of Xinjiang Phytomedicine Resource Utilization, Xinjiang Production and Construction Corps Key Laboratory of Oasis Town and Mountain-basin System Ecology, Shihezi 832000, Xinjiang, China
2. Shihezi Health School, Shihezi 832000, Xinjiang, China

Received:2025-01-10 Revised:2025-03-26 Published:2025-05-15 Online:2025-10-22

摘要/Abstract

摘要： 依托前期55 ℃高温胁迫下耐干苔藓齿肋赤藓的转录组数据，运用实时荧光定量聚合酶链式反应（Quantitative Real time Polymerase Chain Reaction，RT-qPCR)技术，探究9个高温差异表达基因（ScLEA14、ScGSTF11、ScHSP70-17、ScHsfB4b、ScMYB117、ScGLK1、ScERF039、ScERF016、ScbHLH104）在高温、干旱-复水及NaCl胁迫下的表达特征，以此验证转录组测序（RNA sequencing，RNA-Seq）数据的可靠性，为后续齿肋赤藓抗逆基因功能验证提供理论支撑。结果显示：（1） 9个基因在高温胁迫下的表达趋势与RNA-Seq数据基本一致。（2）在极端高温和干旱-复水胁迫时，9个基因均被不同程度诱导，其中3个基因在干旱24 h时诱导至表达峰值，8个基因在复水阶段诱导效果更为突出。（3）在NaCl盐胁迫下，9个耐热相关基因均受盐诱导而上调表达，其中6个基因表现尤为显著。由此得出：ScLEA14、ScMYB117、ScERF016这3个基因在极端高温、干旱-复水和NaCl高盐胁迫下均显著诱导表达，可作为后续抗逆研究的候选基因。

关键词: 耐干苔藓, 齿肋赤藓, 高温胁迫, 转录组, 实时荧光定量PCR

Abstract:

Based on transcriptomic data of the desiccation-tolerant moss Syntrichia caninervis under prior 55 ℃ heat stress, this study employed real-time fluorescent quantitative PCR (RT-qPCR) to investigate the expression patterns of nine heat-responsive differentially expressed genes (ScLEA14, ScGSTF11, ScHSP70-17, ScHsfB4b, ScMYB117, ScGLK1, ScERF039, ScERF016, and ScbHLH104) under high temperature, drought-rehydration, and NaCl stress conditions. The aim was to validate the reliability of RNA-Seq data and provide theoretical support for subsequent functional studies on stress-resistant genes in S. caninervis. Results demonstrated that: (1) The expression profiles of all nine genes under high-temperature stress exhibited substantial concordance with RNA-Seq data, confirming the stability of transcriptomic sequencing. (2) Under extreme heat and drought-rehydration stresses, all genes were differentially induced, with three genes attaining peak expression levels following 24-hour drought treatment, while eight genes displayed more prominent transcriptional activation during the rehydration phase. (3) NaCl stress triggered significant upregulation of all nine thermotolerance-associated genes, with six genes demonstrating statistically robust induction. Thus, the results demonstrate that the three genes ScLEA14, ScMYB117, and ScERF016 are strongly induced under extreme high temperature, drought-rehydration, and NaCl-induced high salinity stress, highlighting their potential as key candidate genes for further investigation into stress resistance mechanisms.

Key words: desiccation moss, Syntrichia caninervis, high temperaturestress, transcriptome, RT-qPCR

霍雯婷, 顾天麒, 高梦宇, 宋艳芳, 李鸿彬, 卓露. 基于RT-qPCR技术解析齿肋赤藓耐热基因在不同非生物胁迫下的表达模式[J]. 干旱区研究, 2025, 42(5): 885-894.

HUO Wenting, GU Tianqi, GAO Mengyu, SONG Yanfang, LI Hongbin, ZHUO Lu. Analysis of the expression patterns of the heat-resistant gene of Syntrichia caninervis under different abiotic stresses based on RT-qPCR technology[J]. Arid Zone Research, 2025, 42(5): 885-894.

图/表 5

表1

表2

图1

图2

图3

参考文献 36

[1]	陈颖, 邵伟玲, 曹萌, 等. 新疆夏季高温日数的变化特征及其影响因子[J]. 干旱区研究, 2020, 37(1): 58-66.
	[Chen Ying, Shao Weiling, Cao Meng, et al. Variation characteristics of summer high-temperature days in Xinjiang and their influencing factors[J]. Arid Zone Research, 2020, 37(1): 58-66.]
[2]	刘璐, 刘普幸, 张旺雄, 等. 1961—2017年新疆极端暖事件变化特征及其未来情景预估[J]. 干旱区研究, 2021, 38(6): 1590-1600. doi: 10.13866/j.azr.2021.06.11
	[Liu Lu, Liu Puxing, Zhang Wangxiong, et al. Variation characteristics of extreme warm events in Xinjiang from 1961 to 2017 and their future projections[J]. Arid Zone Research, 2021, 38(6): 1590-1600.]
[3]	林晓华, 卓一林, 柳丽芳, 等. NaCl胁迫对荒漠苔藓齿肋赤藓膜结构稳定性的影响[J]. 生态学报, 2024, 44(8): 3483-3491.
	[Lin Xiaohua, Zhuo Yilin, Liu Lifang, et al. Effects of NaCl stress on membrane structural stability in the desert moss Syntrichia caninervis[J]. Acta Ecologica Sinica, 2024, 44(8): 3483-3491.]
[4]	Cao T, Haxim Y, Liu X J, et al. ScATG8 gene cloned from desert moss Syntrichia caninervis exhibits multiple stress tolerance[J]. Plants, 2024, 13(1): 59.
[5]	聂华丽, 吴楠, 梁少民, 等. 不同沙埋深度对刺叶墙藓植株碎片生长的影响[J]. 干旱区研究, 2006, 23(1): 66-70.
	[Nie Huali, Wu Nan, Liang Shaomin, et al. Effects of different sand burial sepths on the growth of Bryum argenteum fragments[J]. Arid Zone Research, 2006, 23(1): 66-70.]
[6]	马瑞. 荒漠齿肋赤藓抗旱基因ScALDH21、Sc288对干旱胁迫的响应[D]. 长沙: 中南林业科技大学, 2011.
	[Ma Rui. Response of Drought Resistance Genes ScALDH21 and Sc288 in Desert Moss Syn-trichia caninervis to Drought Stress[D]. Changsha: Central South University of Forestry and Technology, 2011.]
[7]	杨红兰, 张道远, 刘燕, 等. 齿肋赤藓乙醛脱氢酶基因ALDH21的克隆与表达分析[J]. 基因组学与应用生物学, 2010, 29(1): 24-30.
	[Yang Honglan, Zhang Daoyuan, Liu Yan, et al. Cloning and expression analysis of the aldehyde dehydrogenase gene ALDH21 in Syntrichia caninervis[J]. Genomics and Applied Biology, 2010, 29(1): 24-30.]
[8]	Liang Y, Li X, Yang R, et al. BaDBL1, a unique DREB gene from desiccation tolerant moss Bryum argenteum, confers osmotic and salt stress tolerances in transgenic Arabidopsis[J]. Plant Sciences, 2021, 313: 111047.
[9]	韩志立, 尹本丰, 杨孜悦, 等. 积雪变化对温带荒漠齿肋赤藓结皮土壤磷组分的影响[J]. 生态学报, 2024, 44(16): 7119-7129.
	[Han Zhili, Yin Benfeng, Yang Ziyue, et al. Effects of snow cover change on phosphorus fractions in syntrichia caninervis crust soil in temperate deserts[J]. Acta Ecologica Sinica, 2024, 44(16): 7119-7129.]
[10]	Li X, Bai W, Yang Q, et al. The extremotolerant desert moss Syntrichia caninervis is a promising pioneer plant for colonizing extraterrestrial environments[J]. The Innovation, 2024, 5(4): 100657.
[11]	Mao Y, Liu W, Yang X, et al. Syntrichia caninervis adapt to mercury stress by altering submicrostructure and physiological properties in the Gurbantünggüt Desert[J]. Scientific Reports, 2022, 12(1): 11717.
[12]	Bai W, Salih H, Yang R, et al. ScDREBA5 enhances cold tolerance by regulating photosynthetic and antioxidant genes in the desert moss Syntrichia caninervis[J]. Plant, Cell & Environment, 2025, 48(5): 3293-3313.
[13]	Salih H, Bai W, Liang Y, et al. ROS scavenging enzyme-encoding genes play important roles in the desert moss Syntrichia caninervis response to extreme cold and desiccation stresses[J]. International Journal of Biological Macromolecules, 2024, 254: 127778.
[14]	Li H, Zhang D, Li X, et al. Novel DREB A-5 subgroup transcription factors from desert moss (Syntrichia caninervis) confers multiple abiotic stress tolerance to yeast[J]. Journal of Plant Physiology, 2016, 194: 45-53.
[15]	Li X, Yang R, Liang Y, et al. The ScAPD1-like gene from the desert moss Syntrichia caninervis enhances resistance to Verticillium dahliae via phenylpropanoid gene regulation[J]. The Plant Journal, 2023, 113(1): 75-91.
[16]	Li X, Zhang D, Gao B, et al. Transcriptome-wide identification, classification, and characterization of AP2/ERF family genes in the desert moss Syntrichia caninervis[J]. Frontiers in Plant Science, 2017, 8:262.
[17]	Liang Y, Li X, Zhang D, et al. ScDREB8, a novel A-5 type of DREB gene in the desert moss Syntrichia caninervis, confers salt tolerance to Arabidopsis[J]. Plant Physiol Biochem, 2017, 120: 242-251.
[18]	Li X, Liang Y, Gao B, et al. ScDREB10, an A-5c type of DREB gene of the desert moss Syntrichia caninervis, confers osmotic and salt tolerances to Arabidopsis[J]. Genes, 2019, 10(2): 146.
[19]	Yang R, Li X, Yang Q, et al. Transcriptional profiling analysis providing insights into desiccation tolerance mechanisms of the desert moss Syntrichia caninervis[J]. Frontiers in Plant Science, 2023, 14: 1127541.
[20]	Silva A T, Gao B, Fisher K M, et al. To dry perchance to live: Insights from the genome of the desiccation-tolerant biocrust moss Syntrichia caninervis[J]. The Plant Journal, 2021, 105(5): 1339-1356.
[21]	薛山. 新疆特色沙漠植物齿肋赤藓热胁迫响应关键基因的筛选与分析[D]. 石河子: 石河子大学, 2024.
	[Xue Shan. Screening and Analysis of Key Genes Responding to Heat Stress in the Xinjiang Desert Plant Syntrichia caninervis[D]. Shihezi: Shihezi University, 2024.]
[22]	梁玉青, 李小双, 高贝, 等. 基于RNA-Seq数据筛选的银叶真藓耐干相关基因表达模式研究[J]. 植物生理学报, 2017, 53(3): 388-396.
	[Liang Yuqing, Li Xiaoshuang, Gao Bei, et al. Study on the expression patterns of desiccation-tolerance related genes in Bryum argenteum based on RNA-Seq data screening[J]. Journal of Plant Physiology, 2017, 53(3): 388-396.]
[23]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT Method[J]. Methods, 2001, 25(4): 402-408. doi: 10.1006/meth.2001.1262 pmid: 11846609
[24]	宋士伟, 焦德志, 陈旭, 等. 野大麦对干旱胁迫的生理响应与转录组分析[J]. 干旱区研究, 2019, 36(4): 909-915. doi: 10.13866/j.azr.2019.04.15
	[Song Shiwei, Jiao Dezhi, Chen Xu, et al. Physiological responses and transcriptome analysis of wild barley (Hordeum brevisubulatum) under drought stress[J]. Arid Zone Research, 2019, 36(4): 909-915.] doi: 10.13866/j.azr.2019.04.15
[25]	Hayford R K, Serba D D, Xie S, et al. Global analysis of switchgrass (Panicum virgatum L.) transcriptomes in response to interactive effects of drought and heat stresses[J]. BMC Plant Biology, 2022, 22(1): 107. doi: 10.1186/s12870-022-03477-0 pmid: 35260072
[26]	Ji W, Zhu Y, Li Y, et al. Over-expression of a glutathione S-transferase gene, GsGST, from wild soybean (Glycine soja) enhances drought and salt tolerance in transgenic tobacco[J]. Biotechnology Letters, 2010, 32(8): 1173-1179.
[27]	Xu J, Xing X J, Tian Y S, et al. Transgenic Arabidopsis plants expressing tomato glutathione S-Transferase showed enhanced resistance to salt and drought stress[J]. PLoS One, 2015, 10(9): e0136960.
[28]	Khan A H, Wu Y, Luo L, et al. Proteomic analysis reveals that the heat shock proteins 70-17 and BiP5 enhance cotton male fertility under high-temperature stress by reducing the accumulation of ROS in anthers[J]. Industrial Crops and Products, 2022, 188(B): 115693.
[29]	Magar M M, Liu H, Yan G. Genome-wide analysis of AP2/ERF superfamily genes in contrasting wheat genotypes reveals heat stress-related candidate genes[J]. Frontiers in Plant Science, 2022, 13: 853086.
[30]	Pratyusha D S, Sarada D V L. MYB transcription factors-master regulators of phenylpropanoid biosynthesis and diverse developmental and stress responses[J]. Plant Cell Reports, 2022, 41(12): 2245-2260. doi: 10.1007/s00299-022-02927-1 pmid: 36171500
[31]	Wang F, Ren X, Zhang F, et al. A R2R3-type MYB transcription factor gene from soybean, GmMYB12, is involved in flavonoids accumulation and abiotic stress tolerance in transgenic Arabidopsis[J]. Plant Biotechnology Reports, 2019, 13(3): 219-233.
[32]	Shukla R K, Tripathi V, Jain D, et al. CAP2 enhances germination of transgenic tobacco seeds at high temperature and promotes heat stress tolerance in yeast[J]. The FEBS Journal, 2009, 276(18): 5252-5262.
[33]	Li X, Lu J, Zhu X, et al. AtMYBS1 negatively regulates heat tolerance by directly repressing the expression of MAX1 required for strigolactone biosynthesis in Arabidopsis[J]. Plant Communications, 2023, 4(6): 100675.
[34]	黄畋柳, 张锐, 贺迎骁, 等. 辣椒NAC家族成员鉴定及其编码基因在NaCl胁迫下的表达分析[J]. 植物资源与环境学报, 2023, 32(4): 12-24.
	[Huang Tianliu, Zhang Rui, He Yingxiao, et al. Identification of NAC family members in pepper and expression analysis of their encoding genes under NaCl stress[J]. Journal of Plant Resources and Environment, 2023, 32(4): 12-24.]
[35]	Reddy P S, Kavi Kishor P B, Seiler C, et al. Unraveling regulation of the small heat shock proteins by the heat shock factor HvHsfB2c in barley: Its implications in drought stress response and seed development[J]. PLoS One, 2014, 9(3): e89125.
[36]	Si W, Liang Q, Chen L, et al. Ectopic overexpression of maize heat stress transcription factor ZmHsf05 confers drought tolerance in transgenic rice[J]. Genes, 2021, 12(10): 1568.

基因	引物序列（5′→3′）		产物长度/bp
基因	正向引物	反向引物	产物长度/bp
ScLEA14	ATGATCACGATCTGCCCAT	TTATGCGGCTCCCAACTGA	148
ScGSTF11	GAGTCTCACCAAGTTAAACAGCC	ATCTTCTTTTCCACGTACTGCAA	89
ScHSP70-17	GAAGGACTGAGCATGTTTCCTCT	AGGCCATCTCTGTACAATCAGC	150
ScHsfB4b	CAGTACCAGTTCAAAGGATCACCC	TTCGTTTGGACAATGCAGTCACT	84
ScMYB117	CGGTGCTGCTCTGAAACCAA	CTTGCCTGAAGCACAAACCAC	82
ScGLK1	CCTCCTCCGACCTGTTAGATGT	CGCAACTAATCCCCGATGCC	129
ScERF039	GTTGCATCTGCCGTCATCGAAC	CCCGCTACTACTGTCCTTCGTC	118
ScERF016	AGAAGAATGGGAGTCAGAGAGAAGC	CCTTCACGCTCTTCTTCGCTAC	81
ScbHLH104	TCACAACTGCGGTATCTGGT	AATCGTAGCTTTGTCGGTCT	125
α-TUB	CGGTCATTACACCGTGGGAA	CCTCTCCAGCAACAGCGAA	243

基因分类	基因名称	基因差异表达倍数变化（Log₂）
基因分类	基因名称	10 min	30 min	45 min	60 min
功能基因	ScLEA14	+0.73	+0.98	+1.71	+1.37
	ScGSTF11	+1.59	+2.39	+2.99	+2.66
	ScHSP70-17	+0.67	+0.95	+0.90	+0.77
转录因子	ScHsfB4b	+1.54	+2.27	+2.42	+2.25
	ScMYB117	+1.28	+1.43	+1.58	+1.37
	ScGLK1	+2.45	+2.76	+3.10	+3.31
	ScERF039	+0.67	+0.68	+0.68	+0.70
	ScERF016	+0.76	+1.39	+1.95	+1.75
	ScbHLH104	+1.33	+1.45	+1.51	+1.60

基于RT-qPCR技术解析齿肋赤藓耐热基因在不同非生物胁迫下的表达模式

Analysis of the expression patterns of the heat-resistant gene of Syntrichia caninervis under different abiotic stresses based on RT-qPCR technology

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 5

参考文献 36

相关文章 1

Metrics

本文评价

推荐阅读 0