枸杞FBN基因家族鉴定及与类胡萝卜素代谢的相关性分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0340

摘要/Abstract

摘要：

目的原纤维蛋白（fibrillin, FBN）在植物生长发育及类胡萝卜素积累中发挥着重要作用。分析鉴定宁夏枸杞（Lycium barbarum L.）FBN基因家族，为深入研究LbaFBN基因功能奠定基础。方法利用生物信息学方法分析枸杞FBN家族基因的理化性质、基因结构、保守基序、共线性关系及顺式作用元件等。采用转录组和RT-qPCR分析LbaFBNs基因在‘宁杞1号’不同果实发育时期的表达，使用高效液相色谱法测定果实发育时期类胡萝卜素的含量，应用R软件分析LbaFBN表达与类胡萝卜素含量的相关性。结果在枸杞全基因组范围内共鉴定到13个LbaFBNs家族成员，分为12个亚类，不均匀分布在8条染色体上。同一亚类LbaFBNs成员表现出相似的基因结构和基序组成特征。LbaFBNs启动子区域发现含有与植物生长发育、激素响应及逆境胁迫等相关的作用元件。基因表达分析表明，LbaFBNs在不同发育时期均有表达，LbaFBN13随着果实发育显著上调表达。亚细胞定位表明，LbaFBN13蛋白定位在叶绿体中。相关性分析表明，LbaFBN2/4/10表达与紫黄质和叶黄素的含量为显著正相关，LbaFBN3/8表达与β-隐黄质的含量为显著正相关，LbaFBN13表达与玉米黄质双棕榈酸脂含量为显著正相关。结论从枸杞基因组中鉴定出13个LbaFBNs家族成员，不同家族成员在果实发育中表达模式存在差异。LbaFBN2/3/4/8/10/13基因可能参与枸杞果实类胡萝卜素代谢调控。

关键词: 宁夏枸杞, FBN基因家族, 进化分析, 差异表达, 果实发育时期, 类胡萝卜素代谢, 相关性分析, 亚细胞定位

Abstract:

Objective Fibrillin (FBN) protein family members play important roles in plant growth and development, and carotenoid accumulation. The identification of FBN gene family in Lycium barbarum may provide theoretical basis for studying the functions of LbaFBN genes. Method The physicochemical properties, gene structure,conserved motif, collinearity relationship and cis-acting elements of FBN gene family in L. barbarum were analyzed by bioinformatics method. The expression patterns of LbaFBN genes of ‘Ningqi 1’ were investigated using transcriptome and RT-qPCR. The carotenoid content during fruit development was determined by high-performance liquid chromatography. The correlation between LbaFBN gene expression and carotenoid content was analyzed by R software. Result In total, thirteen LbaFBNs family members were identified in L. barbarum and classified into twelve subgroups, which were randomly distributed on 8 chromosomes. The LbaFBNs in each sub-group shared similar gene structure and conserved motifs. Several cis-acting elements related to plant growth and development, hormone, light and stress were found in the promoter region of LbaFBN members. Gene expression analysis showed that LbaFBNs family genes were expressed in different developmental stages. LbaFBN13 expressions increased gradually as the fruit development. Subcellular localization showed that LbaFBN13 protein was localized in the chloroplast. Correlation analysis indicated that the expressions of LbaFBN2/4/10, LbaFBN3/8 and LbaFBN13 were significantly and positively correlated with the content of violaxanthin and lutein, β-cryptoxanthin, and zeaxanthin dipalmitate, respectively. Conclusion Thirteen LbaFBNs family members are identified in the whole genome of L. barbarum, and the expression patterns of these genes in fruit development are different. LbaFBN2/3/4/8/10/13 gene may be involved in carotenoid metabolism.

Key words: Lycium barbarum L., FBN gene family, evolutionary analysis, differential expression, fruit development period, carotenoids metabolism, correlation analysis, subcellular localization

尹跃, 秦小雅, 米佳, 安巍, 何军, 张锋锋. 枸杞FBN基因家族鉴定及与类胡萝卜素代谢的相关性分析[J]. 生物技术通报, 2026, 42(3): 338-348.

YIN Yue, QIN Xiao-ya, MI Jia, AN Wei, HE Jun, ZHANG Feng-feng. Identification of FBN Gene Family and Its Relationship with Carotenoids Metabolism in Lyciumbarbarum[J]. Biotechnology Bulletin, 2026, 42(3): 338-348.

图/表 12

图1 不同发育时期‘宁杞1号’果实S1：幼果期；S2：青果期；S3：转色期；S4：膨大期；S5：成熟期。下同

Fig. 1 Fruits of 'Ningqi 1' at different developmental stagesS1: Young fruit stage. S2: Green fruit stage. S3: Breaker stage. S4: Expanding stage. S5: Mature stage. The same below

表1 本研究中所用引物的序列信息

Table 1 Primer sequence information used in this study

基因名称 Gene name	上游引物序列 Forward primer sequence （5′‒3′）	下游引物序列 Reverse primer sequence （5′‒3′）	用途 Purpose
LbaFBN1	TCACAAGTGCGGTTCTGCGT	AGGAGCACGTTCAACAATCAGGT	荧光定量 Quantitative real-time
LbaFBN2	TTGAAGGGGAGTGGCAAATGA	CCATTTGCTTTGACAATCTGCTC
LbaFBN3	GTTCGAAGCCCCTCTAGAATACAG	TAATCTTTTGCCCAAACACATCC
LbaFBN4	CAAGAGCGCAAAGGTTCAAATAC	GCATAGGGTGTTGGAGTAAAGCA
LbaFBN5	CTCCTGGAAGAGCTCAAGGTGAG	GAAGGATCAACCATTTTCAAAACG
LbaFBN6	GTGTGATACCCATAAAAAGCCGT	GAATAAACAGGTTTCCTTGGTTGC
LbaFBN8	CTCCAGCAAGTTTTCTGTTGAGG	CAAAGCACCCTTTAGTTGCTCA
LbaFBN9	CCGGGAACTTATCACAGCTGC	CTCCCCTTGTTACACGTGTATCAG
LbaFBN10	GAACTCAAGGTAGGGGGCCTAGA	GCGTCTTTGAAACACAAACAAGG
LbaFBN11	CCTCCTCTCAAGGTTCCAATTCC	TCCTTAACGAGAACAAAAAGCCC
LbaFBN12	TGCAATAACACCAGACCAGTTG	CTTGTCATCTCTGCCTATCCTCG
LbaFBN13	ACCGCCAAAGCCAAAGGAAC	TTGGGTTCTTGGACTCAAGCTGA
LbaActin	CAATCGGGTATTTCAAGGTCAAG	GAGCAGTGTTTCCCAGCATTG
LbaFBN13-GFP	ACGAGCTGTACAAGGGTACCATGG CTTCCATCTCTTCTCTG	GGATCCTCTAGATGAACTAGTTAA GGCTTCAAGAGGGGAC	亚细胞定位 Subcellular localization

表2 枸杞FBN基因家族成员基本信息

Table 2 Basic information of LbaFBN gene family in L. barbarum No.1

基因名称 Gene name	基因ID Gene ID	氨基酸数量 Number of amino acids	相对分子量 Molecular weight （kD）	理论等电点 Theoretical pI	总平均疏水性指数 Grand average of hydropathicity	亚细胞定位 Subcellular location
LbaFBN1	Lba01g00762	216	24.61	7.78	-0.345	叶绿体 Chloroplast
LbaFBN2	Lba02g00336	410	45.99	9.15	-0.208	细胞胞质 Cytoplasmic
LbaFBN3	Lba04g00802	332	36.45	4.73	-0.361	叶绿体 Chloroplast
LbaFBN4	Lba04g01207	699	78.47	8.09	-0.257	细胞核 Nuclear
LbaFBN5	Lba06g00949	335	38.38	9.31	-0.289	叶绿体 Chloroplast
LbaFBN6	Lba06g02827	246	27.63	9.82	-0.299	叶绿体 Chloroplast
LbaFBN7	Lba08g00868	198	22.34	7.54	-0.215	细胞核 Nuclear
LbaFBN8	Lba08g01931	302	33.68	6.78	-0.088	叶绿体 Chloroplast
LbaFBN9	Lba09g00405	279	30.17	6.61	-0.247	叶绿体 Chloroplast
LbaFBN10	Lba09g02050	289	32.20	9.04	-0.271	质膜 Plasma membrane
LbaFBN11	Lba09g02224	378	41.83	4.54	-0.352	叶绿体 Chloroplast
LbaFBN12	Lba10g00821	752	84.03	8.33	0.002	质膜 Plasma membrane
LbaFBN13	Lba12g00894	321	35.37	5.28	-0.342	叶绿体 Chloroplast

图2 枸杞与其他5个物种FBN家族蛋白系统进化树

Fig. 2 Phylogenetic tree of FBN family protein in L. barbarum and other five species

图3 枸杞FBN家族成员进化关系（A）、基因结构（B）和保守基序（C）分析

Fig. 3 Phylogenetic relationship (A), gene structure (B), and conserved motif (C) analysis of LbaFBN family members

图4 枸杞FBN基因的染色体定位（A）、物种内共线性（B）和物种间共线性（C）分析

Fig. 4 Chromosome distribution (A), intraspecific collinearity (B), and interspecies collinearity (C) analysis of FBN genes in L. barbarum

图5 LbaFBN基因启动子顺式作用元件分析图A中方格的不同颜色和数字表示LbaFBNs基因启动子中不同元件的数量；图B中不同颜色的直方图表示每个类别中顺式作用元件的总和

Fig. 5 Analysis of cis-acting elements of LbaFBN genes promotersIn Fig. A, the different colors and numbers of the grids indicate the numbers of different promoter elements in these LbaFBN genes. In Fig. B, the different colored histogram indicates the sum of the cis-acting elements in each category

图6 枸杞FBN家族的蛋白互作网络

Fig. 6 Protein-protein interaction network of FBN family in L. barbarum

图7 LbaFBNs基因在‘宁杞1号’5个果实发育阶段的表达模式A：基于RNA-seq分析的LbaFBN基因的表达模式；B：利用RT-qPCR验证LbaFBN基因表达；不同小写字母表示差异显著（P<0.05）。下同

Fig. 7 Expression profile of LbaFBNs genes in 'Ningqi 1' during five fruit developmental stagesA: Transcript profiling of LbaFBN genes based on RNA-seq analysis. B: Gene expression patterns of LbaFBN genes using RT-qPCR validation. The different letters indicate significant differences at the 0.05 level. The same below

图8 ‘宁杞1号’果实5个发育时期类胡萝卜含量变化DW：干重

Fig. 8 Variations in carotenoid contents in 'Ningqi 1' at five different developmental stagesDW: Dry weight

图9 LbaFBNs基因表达与类胡萝卜素含量的相关性分析*，**，***分别表示在P<0.5、P<0.01和P<0.001水平显著

Fig. 9 Correlation analysis between LbaFBNs gene expression and carotenoids content*, **, and *** indicates significant levels at P<0.5, P<0.01, and P<0.001, respectively

图10 LbaFBN13蛋白的亚细胞定位（标尺为20 μm）

Fig. 10 Subcellular localization of the protein LbaFBN13 (Bar=20 μm)

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