铁皮石斛热激蛋白HSP70家族基因鉴定及温度胁迫下的表达分析

doi:10.16420/j.issn.0513-353x.2020-0710

园艺学报 ›› 2021, Vol. 48 ›› Issue (9): 1743-1754.doi: 10.16420/j.issn.0513-353x.2020-0710

铁皮石斛热激蛋白HSP70家族基因鉴定及温度胁迫下的表达分析

赵溪¹, 张婷婷¹, 邢文婷³, 王健², 宋希强², 周扬¹^,^**()

¹海南大学园艺学院,海南省热带园艺作物品质调控重点实验室,海南省热带特色花木资源生物学重点实验室,海南省耐盐作物生物技术重点实验室,海口 570228
²海南大学林学院,热带特色林木花卉遗传与种质创新教育部重点实验室,海口 570228
³海南省林业科学研究院,海口 571000

收稿日期:2021-03-04 修回日期:2021-05-07 出版日期:2021-09-25 发布日期:2021-09-30
通讯作者: 周扬 E-mail:zhouyang@hainanu.edu.cn
基金资助:
海南省自然科学基金项目(319MS009);海南省自然科学基金项目(318QN189);海南省教育厅项目(Hys2020-242);海南省教育厅项目(Hnky2021-19);海南大学科研启动项目(KYQD（ZR）1845);海南省自然科学基金创新团队项目(2018CXTD331)

Genome-wide Identification and Expression Analysis Under Temperature Stress of HSP70 Gene Family in Dendrobium catenatum

ZHAO Xi¹, ZHANG Tingting¹, XING Wenting³, WANG Jian², SONG Xiqiang², ZHOU Yang¹^,^**()

¹Key Laboratory for Quality Regulation of Tropical Horticultural Plants of Hainan Province / Key Laboratory of Germplasm Resources Biology of Tropical Special Ornamental Plants of Hainan Province / Hainan Key Laboratory for Biotechnology of Salt Tolerant Crops,School of Horticulture,Hainan University,Haikou 570228,China
²Key Laboratory of Ministry of Education for Genetics and Germplasm Innovation of Tropical Special Trees and Ornamental Plants,School of Forestry,Hainan University,Haikou 570228,China
³Hainan Academy of Forestry,Haikou 571000,China

Received:2021-03-04 Revised:2021-05-07 Online:2021-09-25 Published:2021-09-30
Contact: ZHOU Yang E-mail:zhouyang@hainanu.edu.cn

摘要/Abstract

摘要：

为研究铁皮石斛（Dendrobium catenatum Lindl.）应对温度胁迫的响应机制,利用生物信息学方法对其热激蛋白HSP70家族基因进行鉴定。荧光定量PCR技术分析它们的组织表达特性,以及在高温（42 ℃）和低温（4 ℃）胁迫下的表达模式。从其基因组中鉴定出10个家族成员,其编码的氨基酸数在628 ~ 702之间,分子量在69.65 ~ 75.15 kD,理论等电点在5.12 ~ 6.18之间。进化树分析可将这10个蛋白分为4个亚族,且每个亚族蛋白的保守基元相似。基因结构分析表明,HSP70的内含子数量为1 ~ 7不等。HSP70在铁皮石斛中的表达具有组织特异性,在合蕊柱中最高。低温处理铁皮石斛组培苗后,DcHSP70-1和DcHSP70-7呈上调表达,推测其响应低温胁迫;高温胁迫下,DcHSP70-2、DcHSP70-3、DcHSP70-5和DcHSP70-7的表达量上调比较明显,推测它们在响应高温胁迫中起主要作用。

关键词: 铁皮石斛, HSP70家族基因, 特征分析, 温度胁迫, 表达分析

Abstract:

In order to study the resistant mechanism of Dendrobium catenatum response to temperature stress,the heat shock protein 70（HSP70）gene families were identified using bioinformatics methods. Meanwhile,the expression profiles of HSP70 genes in different tissues and under high（42 ℃）and low（4 ℃）temperatures were assayed by quantitative real time PCR（qRT-PCR）method. The results showed that 10 HSP70 genes were identified from D. catenatum,which encode 628 to 702 amino acids, the molecular weight was between 69.65 to 75.15 kD,and the theoretical isoelectric point ranged from 5.12 to 6.18. The HSP70 proteins were divided into four subgroups by phylogenetic tree analysis,and the conserved motifs of each subgroup were similar. Structural analysis showed that the number of introns in HSP70 gene varied from one to seven. Spatial expression pattern analysis showed that HSP70 gene expression was tissue specific and the highest expression was found in gynostemium. The expression levels of DcHSP7-1 and DcHSP70-7 increased after low temperature treatment,which indicated that DcHSP70-1 and DcHSP70-7 responded to cold stress. While the relative expression of DcHSP70-2,DcHSP70-3,DcHSP70-5 and DcHSP70-7 was up-regulated obviously under high temperatuer,then we can speculate that these genes play key roles in response to heat stress.

Key words: Dendrobium catenatum, HSP70 gene family, characteristic analysis, temperature stress, expression analysis

中图分类号:

S682.31

赵溪, 张婷婷, 邢文婷, 王健, 宋希强, 周扬. 铁皮石斛热激蛋白HSP70家族基因鉴定及温度胁迫下的表达分析[J]. 园艺学报, 2021, 48(9): 1743-1754.

ZHAO Xi, ZHANG Tingting, XING Wenting, WANG Jian, SONG Xiqiang, ZHOU Yang. Genome-wide Identification and Expression Analysis Under Temperature Stress of HSP70 Gene Family in Dendrobium catenatum[J]. Acta Horticulturae Sinica, 2021, 48(9): 1743-1754.

图/表 8

参考文献 29

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基因Gene	上游引物（5′-3′）Forward primer	下游引物（5′-3′）Reverse primer
ACTIN	TTGTGTTGGATTCTGGTGATGGTGT	TTTCCCGTTCTGCTGTTGTTGTGAA
DcHSP70-1	ACGAAAGCCCCCCTAAAAC	GCAGGTTTTGAACTGAATGGTC
DcHSP70-2	GTTTGGCAGCACGATAGGGTAG	GCCTATCAACCGCTCCGTATC
DcHSP70-3	TGGTCCCTCGTTCAAGTCCG	CCTCTGCCCCTCTGCGTTAGT
DcHSP70-4	GAGAACTGCCTGTGAGAGAGCG	CGGAGGCACTTCTCGACAGG
DcHSP70-5	CTCGTTCATTCAGGAAACCATTGTC	CAACCCTCAGATGCCCGCTA
DcHSP70-6	GCAGTGACAAACCCAACAAATAC	AAACAGATTTACCGAGATAGGCTTC
DcHSP70-7	TTCTCTACCTACTCCGACAACCAAC	CTTTTCATTAGTGATGGTGATTTTGTTC
DcHSP70-8	AGTTGGGCACAGTCATTGGTATT	GGTTGCCCTGGTCATTTGCT
DcHSP70-9	CAAGGAGCCAAACAAGGGAGT	ACACCACCCACAGTCTCAATACC
DcHSP70-10	TACTCCACAATCACTCGTGCCAG	CAGGCACTTCTCCACAGGTTCC

基因Gene	上游引物（5′-3′）Forward primer	下游引物（5′-3′）Reverse primer
ACTIN	TTGTGTTGGATTCTGGTGATGGTGT	TTTCCCGTTCTGCTGTTGTTGTGAA
DcHSP70-1	ACGAAAGCCCCCCTAAAAC	GCAGGTTTTGAACTGAATGGTC
DcHSP70-2	GTTTGGCAGCACGATAGGGTAG	GCCTATCAACCGCTCCGTATC
DcHSP70-3	TGGTCCCTCGTTCAAGTCCG	CCTCTGCCCCTCTGCGTTAGT
DcHSP70-4	GAGAACTGCCTGTGAGAGAGCG	CGGAGGCACTTCTCGACAGG
DcHSP70-5	CTCGTTCATTCAGGAAACCATTGTC	CAACCCTCAGATGCCCGCTA
DcHSP70-6	GCAGTGACAAACCCAACAAATAC	AAACAGATTTACCGAGATAGGCTTC
DcHSP70-7	TTCTCTACCTACTCCGACAACCAAC	CTTTTCATTAGTGATGGTGATTTTGTTC
DcHSP70-8	AGTTGGGCACAGTCATTGGTATT	GGTTGCCCTGGTCATTTGCT
DcHSP70-9	CAAGGAGCCAAACAAGGGAGT	ACACCACCCACAGTCTCAATACC
DcHSP70-10	TACTCCACAATCACTCGTGCCAG	CAGGCACTTCTCCACAGGTTCC

基因名称 Gene name	基因ID Gene ID	基因组位置 Genome location	氨基酸长度/aa Length of amino acid	分子量/kD Molecular weight	等电点 Isoelectric point
DcHSP70-1	LOC110096241	NW_021318618.1:11670942..11679212	677	73.15	5.96
DcHSP70-2	LOC110099937	NW_021318796.1:2966318..2969515	646	70.92	5.30
DcHSP70-3	LOC110105867	NW_021319178.1:303111..323012	702	74.69	5.25
DcHSP70-4	LOC110108419	NW_021319683.1:1339434..1342555	647	71.07	5.17
DcHSP70-5	LOC110109555	NW_021416236.1:267770..301553	701	75.15	5.31
DcHSP70-6	LOC110110271	NW_021318471.1:2515516..2521212	675	72.63	6.18
DcHSP70-7	LOC110114126	NW_021318595.1:11412740..11416597	644	71.19	5.30
DcHSP70-8	LOC110114354	NW_021318608.1:56084..60615	633	73.40	5.19
DcHSP70-9	LOC110115161	NW_021319682.1:11684596..11689365	628	69.65	5.18
DcHSP70-10	LOC110115835	NW_021318777.1:646156..650108	647	71.09	5.12

基因名称 Gene name	基因ID Gene ID	基因组位置 Genome location	氨基酸长度/aa Length of amino acid	分子量/kD Molecular weight	等电点 Isoelectric point
DcHSP70-1	LOC110096241	NW_021318618.1:11670942..11679212	677	73.15	5.96
DcHSP70-2	LOC110099937	NW_021318796.1:2966318..2969515	646	70.92	5.30
DcHSP70-3	LOC110105867	NW_021319178.1:303111..323012	702	74.69	5.25
DcHSP70-4	LOC110108419	NW_021319683.1:1339434..1342555	647	71.07	5.17
DcHSP70-5	LOC110109555	NW_021416236.1:267770..301553	701	75.15	5.31
DcHSP70-6	LOC110110271	NW_021318471.1:2515516..2521212	675	72.63	6.18
DcHSP70-7	LOC110114126	NW_021318595.1:11412740..11416597	644	71.19	5.30
DcHSP70-8	LOC110114354	NW_021318608.1:56084..60615	633	73.40	5.19
DcHSP70-9	LOC110115161	NW_021319682.1:11684596..11689365	628	69.65	5.18
DcHSP70-10	LOC110115835	NW_021318777.1:646156..650108	647	71.09	5.12

铁皮石斛热激蛋白HSP70家族基因鉴定及温度胁迫下的表达分析

Genome-wide Identification and Expression Analysis Under Temperature Stress of HSP70 Gene Family in Dendrobium catenatum

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铁皮石斛热激蛋白HSP70家族基因鉴定及温度胁迫下的表达分析

Genome-wide Identification and Expression Analysis Under Temperature Stress of HSP70 Gene Family in Dendrobium catenatum

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