马铃薯LAC基因家族的鉴定及盐胁迫下表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0337

摘要/Abstract

摘要：

目的漆酶（laccase, LACs）是一类含有铜离子的多酚氧化酶，在植物生长发育和胁迫响应过程中发挥重要作用。鉴定马铃薯 StLAC 基因家族成员并分析其表达特征，为 StLAC 功能研究提供参考。方法基于马铃薯基因组数据，运用生物信息学方法对马铃薯 StLAC 基因家族进行鉴定；利用转录组数据分析 StLAC 基因在不同组织内和非生物胁迫下的表达模式，并通过 RT-qPCR对其在盐胁迫下表达模式进行验证。结果在马铃薯中鉴定到77个 StLAC 基因，其中56个 StLAC 蛋白质等电点大于7，仅 StLAC 69为疏水蛋白，其余76个成员为亲水蛋白。亚细胞定位预测结果表明，20个 StLAC蛋白定位于叶绿体，19个定位于液泡，其他StLAC蛋白定位在细胞外、细胞质、细胞核和质膜等部位。顺式作用元件分析显示，StLAC 基因家族启动子区域含有光响应、生长发育、激素和胁迫响应等作用元件。转录组数据分析结果表明，StLAC 基因在不同组织内特异性表达，在盐、干旱、激素胁迫下被诱导表达。通过RT-qPCR对12个 StLAC 家族成员（StLAC4/12/17/26/36/40/43/45/55/56/66/68）在盐胁迫下根和叶中的表达模式进行验证，结果显示，胁迫后 StLAC4、StLAC56在根中的表达量上调3.8-15倍，StLAC12、StLAC40、StLAC56在叶中的表达量上调17-34倍。结论马铃薯 StLAC 基因家族成员众多，分布多样，可能在马铃薯响应盐胁迫过程中发挥重要作用。

关键词: 马铃薯, 漆酶, 盐胁迫, 生物信息学分析, 表达分析

Abstract:

Objective Laccases (LACs) are a class of copper-containing polyphenol oxidases that play significant roles in plant growth, development, and stress responses. The aim of this study is to identify the members of the StLAC gene family in potato (Solanum tuberosum L.) and analyze their expression characteristics, providing a reference for the functional study of StLACs. Method Based on the potato genome data, the StLAC gene family in potato was identified using bioinformatics methods. The expression patterns of StLAC genes in different tissues and under abiotic stress were analyzed using transcriptome data, and the expression patterns of StLAC genes under salt stress were verified by RT-qPCR. Result A total of 77 StLAC genes were identified in potato. Among them, 56 StLAC proteins had an isoelectric point >7, and only StLAC69 was a hydrophobic protein, while the other 76 members were hydrophilic proteins. Subcellular localization predicted results indicated that 20 StLAC proteins were located in the chloroplast, 19 in the vacuole, and the remaining StLAC proteins were located in the extracellular space, cytoplasm, nucleus, and plasma membrane. Analysis of cis-acting elements showed that the promoter regions of the StLAC gene family contained elements related to light response, growth and development, hormones, and stress response. Transcriptome data analysis revealed that StLAC genes were specifically expressed in different tissues and were induced under salt, drought, and hormone stress. The expression patterns of 12 StLAC family members (StLAC4/12/17/26/36/40/43/45/55/56/66/68) in the roots and leaves under salt stress were verified by RT-qPCR. The results showed that the expressions of StLAC4 and StLAC56 in the roots were upregulated by 3.8 to 15 times after stress, and the expressions of StLAC12, StLAC40, and StLAC56 in the leaves were upregulated by 17 to 34 times. Conclusion The StLAC gene family in potato is large and diverse, and may play an important role in the response of potato to salt stress.

Key words: potato, laccase, salt stress, bioinformatics analysis, expression analysis

巩慧玲, 邢玉洁, 马俊贤, 蔡霞, 冯再平. 马铃薯LAC基因家族的鉴定及盐胁迫下表达分析[J]. 生物技术通报, 2025, 41(9): 82-93.

GONG Hui-ling, XING Yu-jie, MA Jun-xian, CAI Xia, FENG Zai-ping. Identification of Laccase （LAC） Gene Family in Potato （Solanum tuberosum L.） and Its Expression Analysis under Salt Stresses[J]. Biotechnology Bulletin, 2025, 41(9): 82-93.

图/表 9

表1 RT-qPCR引物信息

Table 1 Primer information for RT-qPCR

基因 Gene	上游引物 Forward primer（5'-3'）	下游引物 Reverse primer（5'-3'）
StLAC4	GTCATTTGGATGTTCACTTGCCTTG	TCTGGTGGAGGAGGAGGTAGC
StLAC12	AGGTCCAGTTGTTTATGCTCAAGAAG	ACACGATAGCCTCTGACGGATG
StLAC17	GGTTGTTCCATTGCCACATTGATTC	CGGTTCCATTGTCTGTTTCTCTCC
StLAC26	AAGGTGATCGTCTTGAAATTGAAGTTG	GTGTTACATAGGCTGGTCCATCTG
StLAC36	ACCTTTGGCACCTCTTTGGAATG	GAGTAGGAGGCTTAGGAGACAACC
StLAC40	TGTCGTTTGTGCCACTTGATACTC	CCTGTATGCTTCCAACTCTGAATCC
StLAC43	ATCGTGTCTGCTACATTGCTCTG	CAAGAGGAGAAGCGGTGATATAAGAG
StLAC45	TGCTATCTTCTCTGCTTCACAACAC	TTCCAATCACCTGCTGCTTAACTC
StLAC55	GGGAAACACCGTTACGCCATC	CACCTTGTCCGCAACACCATAG
StLAC56	AGGCATATAATAGCGTAGTTGGAGTTG	GGGAGGATGGGTGGTGATGAAG
StLAC66	TGTCGTTTGTGCCACTTGATACTC	CCTGTATGCTTCCAACTCTGAATCC
StLAC68	TGCTATCTTCTCTGCTTCACAACAC	TTCCAATCACCTGCTGCTTAACTC

图1 马铃薯（St）和拟南芥（At）LAC基因的系统进化树

Fig. 1 Phylogenetic tree analysis of LAC genes from S. tuberosum (St) and A. thaliana (At)

图2 马铃薯StLAC基因家族的进化树（A）、保守基序结构（B）和基因结构（C）

Fig. 2 Phylogenetic tree (A), conserved motif structure(B), and gene structure(C) in StLAC gene family

图3 马铃薯 StLAC 基因家族成员在染色体上的分布

Fig. 3 Chromosome location of the StLAC gene family in S. tuberosum

图4 拟南芥（At）与马铃薯（St） LAC 基因的染色体共线性关系

Fig. 4 Chromosomal collinear relationship between A. thaliana (At) and S. tuberosum (St) LAC genes

图5 StLAC 家族成员蛋白互作网络

Fig. 5 Protein interactions network among members of the StLAC family

图6 StLAC 基因家族启动子中顺式作用元件的种类和数量

Fig. 6 Type and number of cis-acting elements in the promoter of StLAC gene family

图7 马铃薯 LAC 基因在不同组织中（A）和不同胁迫下（B）的表达分析

Fig. 7 Expression analysis of LAC gene family of S.tuberosum in different tissues (A) and under different stress (B)

图8 马铃薯根（A）和叶（B）中StLAC基因在盐胁迫下的相对表达量数据为3个独立生物学重复的平均值（± SE），不同字母表示P < 0.05时差异显著

Fig. 8 Relative expressions of StLACs in S. tuberosum roots (A) and leaf (B) under salt stressData are mean (± SE) from three independent biological replicates. Different letters indicate significant differences at P < 0.05

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