三七脂氧合酶的全基因组鉴定及其对茉莉酸甲酯和创伤的响应

doi:10.13560/j.cnki.biotech.bull.1985.2025-0726

摘要/Abstract

摘要：

目的鉴定三七LOX基因家族成员并分析其表达特征，为后续深入探究PnLOX家族生物学功能奠定基础。方法通过生物信息学方法鉴定了三七LOX基因家族，并分析了PnLOX基因的系统发育树、基因结构、启动子顺式作用元件及不同组织、MeJA处理、损伤处理下的表达模式。结果从三七基因组中共鉴定出10个候选的PnLOX基因，根据其系统发育树和底物特异性将其分为9S-LOX、SG I 13S-LOX和SG II 13S-LOX三个亚组，并发现进化相近的LOX基因具有相似的基因结构和保守基序。此外，PnLOX基因定位于6条染色体上，并在进化过程中通过片段复制（SD）和串联重复（TD）产生。亚细胞定位预测发现除SG I 13S-LOX亚组的PnLOX1/2/9定位于叶绿体外，其他所有PnLOX蛋白均定位于细胞质中。不同组织表达分析发现PnLOX基因在不同组织中差异表达。物理损伤和外源MeJA处理发现PnLOX1和PnLOX2基因在MeJA处理和物理损伤后快速激活。结论从三七基因组中鉴定了10个候选的PnLOX基因，其中PnLOX1和PnLOX2基因可能在三七JA合成及JA介导的损伤修复中发挥作用。

关键词: 三七, 脂氧合酶（LOX）, 基因家族, 损伤, 茉莉酸（JA）

Abstract:

Objective To identify the Panax notoginsengLOX gene family and analyze its expression characteristics, thereby providing a basis for further investigation into the biological functions of the PnLOX genes. Method Bioinformatics methods were used to identify P. notoginseng LOX gene family and analyze its the phylogenetic relationship, gene structure characteristics, cis-elements and expression patterns of different plant tissues, MeJA treatment and wounding stress. Result Ten putative PnLOX genes were identified and formed three subgroups (9S-LOX, SG I 13S-LOX and SG II 13S-LOX ) according to phylogenetic tree and LOX substrate specificity. It was observed that LOX genes with similar evolutionary relationships had similar gene structures and conserved motifs. The PnLOX genes was located on six chromosomes and expanded mainly via segmental duplication (SD) and tandem duplication (TD). Subcellular localization prediction revealed that, except for PnLOX1/2/9 from SG I 13S-LOX subgroup, which localized to the chloroplast, all other PnLOX localized to the cytoplasm. Tissue expression analysis indicated that the PnLOX genes were differentially expressed in different tissues. The PnLOX1 and PnLOX2 genes were rapidly activated after MeJA treatment and wounding. Conclusion Ten putative PnLOX genes are identified from the P. notoginseng genome, among which the PnLOX1 and PnLOX2 genes may play an essential role in JA biosynthesis and JA-mediated damage repair in P. notoginseng.

Key words: Panax notoginseng, lipoxygenases (LOX), gene family, wounding, jasmonic acid (JA)

李健斌, 侯家娥, 李雷林, 艾明涛, 刘天泰, 崔秀明, 杨千. 三七脂氧合酶的全基因组鉴定及其对茉莉酸甲酯和创伤的响应[J]. 生物技术通报, 2026, 42(1): 218-229.

LI Jian-bin, HOU Jia-e, LI Lei-lin, AI Ming-tao, LIU Tian-tai, CUI Xiu-ming, YANG Qian. Genome-wide Identification of Panax notoginseng Lipoxygenases Coupled in Response to Methyl-jasmonate and Wounding[J]. Biotechnology Bulletin, 2026, 42(1): 218-229.

图/表 8

表1 RT-qPCR引物序列

Table 1 Primer sequences for RT-qPCR

基因名 Gene name	正向引物 Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）	产物大小 Product （bp）
PnACTIN2	TCCAAGGGTGAATATGATGAATCG	AACCTCTCCAAAGAGAATTTCTGAGT	199
PnLOX1	TAGAGGGCCTTCCTGCTGAT	GCTTGGAGCTCCCTGTCATT	208
PnLOX2	CCCAATGTGGTCAGCAGGAT	TGGGCCATATATCTCCGGGT	207
PnLOX3	GAGGGATGGCAGTTGAGGAC	CATGGCCCTTTTCTCGGACT	210
PnLOX4	CACTCCAGCTGAAGAGGGTG	GCGGAAGTGAGGTTGGAGAA	199
PnLOX5	CCTTTATCTCCGGCGATGCT	AACCTTGGTGGCCTTATCGG	158
PnLOX6	ACCGACAACTAGGTGCGATG	CTGCTGGCAATCCTTCCTGA	210
PnLOX7	CCTTGACCCTGTGAGCTACG	GTTTTCCTTTTCCCGGTGGC	269
PnLOX8	GCAGGTGGAAGCGTTGAAAG	AACTTCAGGCCATAGGGTGC	167
PnLOX9	AAGCAGGGGTCAGTAACAGC	AGATCCCCTGACCAATCCCA	244
PnLOX10	TCGCCTCCATGAGTTTCCAC	AGGTAGAGGAGTGTGCGACT	234

图1 三七、拟南芥和水稻LOX蛋白的系统发育树不同颜色的形状表示不同物种；不同颜色的扇环表示不同亚组

Fig. 1 Phylogenetic tree of LOX proteins of P. notoginseng, A. thaliana, and Oryza sativaDifferent species are indicated in different shapes and colors. The various colors indicate the different subgroups

图2 PnLOXs的系统发育树、保守基序、保守结构和基因结构域A：PnLOXs系统发育树；B：PnLOXs保守基序；C：PnLOXs保守结构域；D：PnLOXs基因结构；E. PnLOXs保守基序的氨基酸序列

Fig. 2 Phylogenetic relationship, conserved motifs, conserved domain and gene structure of the PnLOXsA: Phylogenetic tree of PnLOXs. B: Conserved motifs of PnLOXs. C: Conserved domain of PnLOXs. D: Gene structure of PnLOXs. D: Amino acid sequence of PnLOXs conserved motifs

图3 PnLOX基因染色体定位和共线性分析

Fig. 3 Chromosomal localization and collinearity analysis of PnLOX gene

图4 PnLOX基因启动子顺式作用元件A：PnLOX基因顺式作用元件在启动子上的分布；不同的颜色代表不同的顺式作用元件；B：PnLOX基因顺式作用元件统计图；网格中的热图和柱状图示顺式作用元素的数量

Fig. 4 Cis-acting elements in the promoter regions of PnLOX genesA: Distribution of PnLOX genes cis-acting elements on the promoter. Different colors indicate the different cis-acting elements. B: The number of cis-acting elements in PnLOX genes. The heatmap in grid and the color columns indicate the numbers of cis-acting elements

图5 PnLOX基因在不同组织中的表达模式不同字母表示差异显著（P<0.05），下同

Fig. 5 Expression profiles of PnLOXs in different tissueDifferent letters indicate significant differences (P<0.05). The same below

图6 PnLOX基因在MeJA处理后的表达模式A：PnLOX基因在MeJA处理后的转录组表达热图；B：6个PnLOX基因的RT-qPCR结果

Fig. 6 Expression profiles of PnLOXs under MeJA treatmentA. Transcriptome expression heatmap of PnLOX genes after MeJA treatment. B. RT-qPCR results of 6 PnLOX genes after MeJA treatment

图7 PnLOX基因在损伤胁迫后的表达模式A：PnLOX基因在损伤后不同时间的表达水平；B：损伤前后的三七叶片

Fig. 7 Expression profiles of PnLOXs in response to wound stressA: Expressions of PnLOX genes at different times after wound stress. B: Un-wounded and wounded leaf of P. notoginseng

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