Identification of the bHLH Gene Family in Tricholoma matsutake and Its Expression Analysis under Heavy Metal Stress

doi:10.15933/j.cnki.1004-3268.2025.07.008

Abstract

Abstract: The bHLH（basic helix‑loop‑helix）transcription factors are widely involved in plant growth，development and stress response. In order to clarify the detailed information of the bHLH family members in Tricholoma matsutake and their response to heavy metal stress，we identified the bHLH family members based on Tricholoma matsutake genome by using bioinformatics methods.The physicochemical properties and structure of the proteins，the conserved domains and the promoter cis‑acting elements of the genes were analyzed，and the expression patterns of the bHLH family members in different developmental stages of Tricholoma matsutake fruit bodies，for different refrigeration durations，and under different heavy metal stresses were also investigated.The results showed that a total of 12 bHLH genes were identified in the Tricholoma matsutake genome，which were evenly distributed on 9 chromosomes.Physicochemical property analyses showed that the number of amino acids，theoretical molecular weight of protein，isoelectric point，and instability coefficient encoded by the bHLH（TmabHLH）genes were in the ranges of 96—878，10.72—96.02 ku，4.72—10.81，and 33.55—89.90，respectively.Subcellular localization analyses showed that all the genes were localized in the nucleus and were mainly hydrophilic and unstable nuclear proteins.In addition，the tertiary structure of TmabHLH protein mainly consisted of α‑helices and irregular convolutions，and the promoter sequences of the TmabHLH genes contained a large number of light‑responsive elements，hormone‑responsive elements，and abiotic stress‑responsive elements，suggesting that they may be involved in the growth and development and stress response of Tricholoma matsutake.Transcriptome analysis showed that most of the TmabHLH genes were abundantly expressed in the early development of Tricholoma matsutake fruit bodies，while most of the TmabHLH genes showed a decreasing trend in expression with prolonged cold storage time.The qRT‑PCR assay revealed that all six TmabHLH genes responded to the four heavy metal stresses and had different expression patterns at different concentrations，e.g.，the genes responded more drastically under arsenic（As）stress than under mercury（Hg），cadmium（Cd）and lead（Pb）stress.In summary，the Tricholoma matsutake bHLH gene family has the function of regulating the response of matsutake to heavy metal stress，which lays an important foundation for further revealing the specific functions of TmabHLHs under heavy metal stress.

Key words: Tricholoma matsutake, bHLH gene family, Abiotic stress, Heavy metal, Expression pattern

摘要： bHLH（Basic helix‑loop‑helix）转录因子广泛参与植物的生长发育及胁迫应答，为明确松茸中bHLH家族成员的详细信息及其对重金属胁迫应答情况，基于松茸基因组，利用生物信息学方法鉴定松茸中bHLH家族成员，并对其蛋白质理化性质及结构、保守结构域、基因启动子顺式作用元件等进行了分析，同时对松茸子实体不同发育阶段、不同冷藏时间及不同重金属胁迫下bHLH家族成员的表达模式进行了研究。结果表明，在松茸基因组中共鉴定到12个bHLH基因，在9条染色体上均匀分布。理化性质分析表明，松茸bHLH（TmabHLH）基因编码氨基酸数目、蛋白质理论分子质量、等电点、不稳定系数分别在96~878个、10.72~96.02 ku、4.72~10.81、33.55~89.90；亚细胞定位分析显示，所有基因均定位于细胞核中，且以亲水、不稳定的核蛋白为主。此外，TmabHLH蛋白的三级结构主要由α-螺旋及无规则卷曲组成，并且TmabHLH 基因的启动子序列中含有大量的光响应元件、激素响应元件和非生物胁迫响应元件，暗示其可能参与松茸的生长发育和胁迫应答。转录组分析显示，大部分TmabHLH 基因在松茸子实体发育早期大量表达，大部分TmabHLH 基因随着冷藏时间的延长表达量呈下降趋势。qRT-PCR检测发现，6个TmabHLH 基因均能响应4种重金属胁迫，并且在不同含量下具有不同的表达模式，如As胁迫下基因的响应程度比Hg、Cd和Pb胁迫下更加剧烈。综上所述，松茸bHLH基因家族具有调控松茸响应重金属胁迫的功能。

关键词: 松茸, bHLH基因家族, 非生物胁迫, 重金属, 表达模式

CLC Number:

S567.3

LANG Qian, ZHANG Min, WANG Hao, GONG Lei, ZHOU Yu. Identification of the bHLH Gene Family in Tricholoma matsutake and Its Expression Analysis under Heavy Metal Stress[J]. Journal of Henan Agricultural Sciences, 2025, 54(7): 72-83.

郎茜, 张敏, 王浩, 龚蕾, 周玉. 松茸bHLH基因家族成员的鉴定及其在重金属胁迫下的表达分析[J]. 河南农业科学, 2025, 54(7): 72-83.

Figures/Tables 12

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