Identification and analysis of heat shock transcription factor gene in <i>Zizania latifolia</i>

doi:10.3785/j.issn.1008-9209.2022.03.281

Journal of Zhejiang University (Agriculture and Life Sciences)

2023, Vol. 49

Issue (2): 269-279 DOI: 10.3785/j.issn.1008-9209.2022.03.281

Horticultural sciences

Identification and analysis of heat shock transcription factor gene in Zizania latifolia

Licong CAI¹(

),Mingjia TANG¹,Jin XU¹,Zhenyu QI²,Feijun FAN³,Yanhong ZHOU¹(

)

^1.College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang, China
^2.Agricultural Experiment Station, Zhejiang University, Hangzhou 310058, Zhejiang, China
^3.Agriculture and Rural Affairs Bureau of Lishui, Lishui 323000, Zhejiang, China

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Abstract

To explore the function and potential applications of heat shock transcription factor (HSF) gene family on heat tolerance of Zizania latifolia of ‘Longjiao No. 2’, twenty-eight HSF proteins were identified and analyzed by bioinformatics methods. The physicochemical analysis showed that the theoretical isoelectric point was 4.77 to 11.63, and the molecular weight was 16.77-101.29 kDa, and the protein length was 239-661 amino acids, and the instability indexes of the whole family were more than 40. The multiple sequence alignment revealed that the DNA-binding domain of HSF protein was highly conserved with a length of about 100 amino acids. The phylogenetic tree of HSF proteins of Z. latifolia, Arabidopsis thaliana, Phyllostachys edulis, and Oryza sativa L. was constructed by MEGA 7.0 and the HSF proteins could be divided into A, B, and C classes. In the HSF family of Z. latifolia, there were 17 members in class A, seven in class B, and four in class C. The expression profiles of HSF genes under the heat stress was analyzed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the physiological indexes under the heat stress were also measured. The results showed that 14 HSF genes were at high expression levels (the expression levels were increased by more than 10 times) after the heat stress (42 ℃, 12 h). Among them, the expression levels of ZlHSF-04, ZlHSF-12, and ZlHSF-27 were up-regulated most obviously, which increased by 37, 36, and 44 times when compared with the normal temperature treatment (CK), respectively. Moreover, under the heat stress, the leaves became dry and curled with a large area of water loss and chlorosis, and the maximum photochemical efficiency (F_v/F_m) of photosystem Ⅱ was significantly reduced by 49.9% compared with the CK. Besides, the relative electrical leakage (REL), and the contents of malondialdehyde (MDA), proline (Pro), hydrogen peroxide (H₂O₂) increased significantly by 409%, 97%, 396%, and 99%, respectively. These results lay theoretical foundations for further study of the functions of the HSF gene family under heat stress of Z. latifolia.

Key words： Zizania latifolia heat stock transcription factor gene family expression analysis

Received: 28 March 2022 Published: 25 April 2023

CLC:

S645.2

Corresponding Authors: Yanhong ZHOU E-mail: 21916121@zju.edu.cn;yanhongzhou@zju.edu.cn

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	Licong CAI
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	Feijun FAN
	Yanhong ZHOU

Cite this article:

Licong CAI,Mingjia TANG,Jin XU,Zhenyu QI,Feijun FAN,Yanhong ZHOU. Identification and analysis of heat shock transcription factor gene in Zizania latifolia. Journal of Zhejiang University (Agriculture and Life Sciences), 2023, 49(2): 269-279.

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https://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2022.03.281 OR https://www.zjujournals.com/agr/Y2023/V49/I2/269

茭白热激转录因子基因的鉴定与分析

为探究热激转录因子（heat shock transcription factor, HSF）基因家族在茭白耐热性中的功能及潜在应用，本试验以‘龙茭2号’为材料，利用生物信息学手段，鉴定出28个茭白HSF蛋白并对其进行分析。理化性质分析表明，茭白HSF家族蛋白的理论等电点为4.77～11.63，分子量为16.77～101.29 kDa，氨基酸长度为239～661个氨基酸，不稳定系数均大于40。多序列比对发现其DNA结合域具有高度保守性，长约为100个氨基酸。通过MEGA 7.0软件构建茭白、拟南芥、毛竹和水稻的HSF蛋白系统发育树，发现HSF蛋白可以分为A、B、C 3个分支，茭白HSF家族中含有17个A类成员、7个B类成员、4个C类成员。进一步通过实时荧光定量聚合酶链反应（real-time fluorescent quantitative polymerase chain reaction, qRT-PCR）分析高温胁迫下茭白HSF基因家族的表达模式和热胁迫生理指标，结果表明，有14个HSF基因在高温胁迫（42 ℃，12 h）后处于高表达水平（表达量提高10倍以上），其中ZlHSF-04、ZlHSF-12、ZlHSF-27表达量上调最明显，与常温对照组（CK）相比分别上调37倍、36倍、44倍；同时，高温胁迫下茭白幼苗叶片出现大面积失水失绿的现象，叶片干枯卷曲，且光系统Ⅱ最大光化学效率较CK降低了49.9%，而相对电导率，丙二醛、脯氨酸、过氧化氢含量与CK相比分别增加了409%、97%、396%和99%。本结果为进一步研究HSF基因家族在茭白热激响应中的功能提供了理论依据。

关键词： 茭白, 热激转录因子, 基因家族, 表达分析

Table 1 Physicochemical properties of HSF family members in Z. latifolia

Fig. 1 Multiple sequence alignment of HSF proteins’ DBDs in Z. latifolia

Fig. 2 Phylogenetic tree of HSF proteins in Z. latifolia, P. edulis, A. thaliana, and O. sativa

Fig. 3 Distribution and types of conserved motifs of HSF family members in Z. latifolia

Fig. 4 Distribution of upstream promoters of HSF family members in Z. latifolia

Fig. 5 Gene structures of HSF family in Z. latifolia

Fig. 6 Expression profiles of HSF gene family in response to heat stress in Z. latifolia

Fig. 7 Effects of heat stress on phenotype (A) and F_v/F_m(B) of seedlings of Z. latifoliaDifferent lowercase letters above bars indicate significant differences at the 0.05 probability level; n=3. The same as below.

Fig. 8 Effects of heat stress on REL (A), MDA content (B), Pro content (C), and H₂O₂ content (D) of leaves in Z. latifolia


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