建兰CNGC基因家族鉴定及其在非生物胁迫下的转录响应

doi:10.16420/j.issn.0513-353x.2024-0952

园艺学报 ›› 2026, Vol. 53 ›› Issue (1): 131-148.doi: 10.16420/j.issn.0513-353x.2024-0952

• 遗传育种·种质资源·分子生物学 • 上一篇下一篇

建兰CNGC基因家族鉴定及其在非生物胁迫下的转录响应

朱炫怡¹, 赵雨晴¹, 唐菲鸿¹, 吴海艳¹, 张春³, 赵凯², 彭东辉¹, 兰思仁¹, 周育真¹^,^*()

¹ 福建农林大学风景园林与艺术学院，兰科植物保护与利用国家林业和草原局重点实验室，海峡花卉产业科技创新高地，福州 350002
² 福建师范大学生命科学学院，福州 350117
³ 漳州市林业科技推广站，福建漳州 363000

收稿日期:2025-03-13 修回日期:2025-09-02 出版日期:2026-01-25 发布日期:2026-01-26
通讯作者:
^*E-mail：zhouyuzhencn@fafu.edu.cn
基金资助:
国家重点研发计划课题(2023YFD1600504); 国家自然科学基金青年项目(32101583)

Whole-Genome Identification of the CNGC Gene Family in Cymbidium ensifolium and Transcriptional Reponses Under Abiotic Stresses

ZHU Xuanyi¹, ZHAO Yuqing¹, TANG Feihong¹, WU Haiyan¹, ZHANG Chun³, ZHAO Kai², PENG Donghui¹, LAN Siren¹, ZHOU Yuzhen¹^,^*()

¹ The Strait Floral Industry Science and Technology Innovation Hub， Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization，College of Landscape Architecture and Art，Fujian Agriculture and Forestry University， Fuzhou 350002， China
² College of Life Sciences， Fujian Normal University， Fuzhou 350117， China
³ The Forestry Science and Technology Extension Station， Zhangzhou， Fujian 363000， China

Received:2025-03-13 Revised:2025-09-02 Published:2026-01-25 Online:2026-01-26

摘要/Abstract

摘要：

为深入了解建兰（Cymbidium ensifolium）CNGC基因的功能，基于建兰全基因组数据对其进行了系统的鉴定和分析，共获得13个包含CNBD、CaMBD和IQ基序的CeCNGC基因，分为5个亚家族，不均匀地分布于10条染色体上，且存在2对片段重复。该家族成员的启动子区域存在大量与激素响应和非生物胁迫密切相关的顺式作用元件，且其调控序列中还包含多个与非生物胁迫响应相关的miRNA结合位点。CeCNGC在不同组织和花发育阶段均有转录，其中CeCNGC3/5/10/11/12/13呈现组织特异性表达模式。干旱、低温和ABA处理下，CeCNGC基因均表现有不同程度转录响应。在干旱胁迫下，CeCNGC1/4/5/8/13/10/12/9/11在处理第3天出现上调，其中CeCNGC1/4/13上调最为显著；而CeCNGC2/3/6/7在第2天出现显著下调。低温胁迫3 ~ 7 d内，CeCNGC2/4/5/7的表达量持续上调。ABA处理下，CeCNGC9/10/12/13的表达水平在第1天显著上调，而CeCNGC1/4/7在第3天显著上调。CeCNGC2在低温和ABA处理下的表达模式一致，且受到强烈转录诱导。

关键词: 建兰, CNGC基因, 表达模式, 非生物胁迫

Abstract:

To further investigate the functional roles of the Cymbidium ensifolium CNGC gene family，a systematic identification and in-depth analysis were conducted utilizing whole-genome data. A total of 13 CeCNGC genes containing CNBD，CaMBD，and IQ motifs were identified，which were classified into five subfamilies. These genes were unevenly distributed across 10 chromosomes，with two pairs of segmental duplications detected. The promoter regions of these genes harbor a large number of cis-acting elements closely associated with hormone responses and abiotic stress regulation. Additionally，their regulatory sequences harbor multiple miRNA binding sites related to abiotic stress responses. Expression analysis revealed that CeCNGC genes were transcribed across different tissues and floral developmental stages，with CeCNGC3/5/10/11/12/13 exhibiting significant tissue-specific transcription patterns. Under drought，low-temperature，and ABA treatments，all CeCNGC genes responded to varying degrees. During drought stress，CeCNGC1/4/5/8/13/10/12/9/11 were up-regulated on the third day，among which CeCNGC1/4/13 exhibited the most significant up-regulation. In contrast，CeCNGC2/3/6/7 were significantly down-regulated on the second day. Under low-temperature stress，the expression levels of CeCNGC2/4/5/7 continuously increased from day 3 to 7. In response to ABA treatment，the expression levels of CeCNGC9/10/12/13 were significantly up-regulated on the first day，whereas CeCNGC1/4/7 were significantly up-regulated on the third day. Notably，CeCNGC2 exhibited a consistent expression pattern under both low-temperature and ABA treatments，with strong induction observed in both conditions.

Key words: Cymbidium ensifolium, CNGC genes, expression patterns, abiotic stress

朱炫怡, 赵雨晴, 唐菲鸿, 吴海艳, 张春, 赵凯, 彭东辉, 兰思仁, 周育真. 建兰CNGC基因家族鉴定及其在非生物胁迫下的转录响应[J]. 园艺学报, 2026, 53(1): 131-148.

ZHU Xuanyi, ZHAO Yuqing, TANG Feihong, WU Haiyan, ZHANG Chun, ZHAO Kai, PENG Donghui, LAN Siren, ZHOU Yuzhen. Whole-Genome Identification of the CNGC Gene Family in Cymbidium ensifolium and Transcriptional Reponses Under Abiotic Stresses[J]. Acta Horticulturae Sinica, 2026, 53(1): 131-148.

图/表 12

表1 荧光定量PCR引物序列 Tbale 1 Primers used for RT-qPCR

引物名称 Primer name	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer
CeCNGC1	AGCCTATCAGACTCCTTCCACCTTG	CGAGCTTCCTTCCTGGCACTTG
CeCNGC2	CTTCGTAATCCTTCCGCTTCCTCAG	GGTACACCTTCGGCAAGAACTGG
CeCNGC3	AACGGTGAAGGCATTATCTGAGGTG	GCTTGCTATGGAGTCTGCGGAAC
CeCNGC4	AGATGCGATATGCGAGCGATTAGTC	TCGTCCACTGGGTCACCTTCC
CeCNGC5	ACAGTGAAGGCATTGTCGGAAGTC	TGAAGGTGTGCTGGAGTTGCTTAC
CeCNGC6	AGTACAAGTGGTTGGCAACTCAAGG	GAGGCAGAGATGTCGCTTAATGTCC
CeCNGC7	CAGGTTGTTCTATGGGTGGCAGTAC	CGGAGGAAGGAGATGGAGTGGTAG
CeCNGC8	GTGGGAGCGGGCACTTTGATG	AGAGAAGCAAGCACTGGCATTAGG
CeCNGC9	CACAGAGAGCATCTTCGTCGTCAG	TCGCCGCAGAAATCGCCTTG
CeCNGC10	CACCGAGAGCACTTACATTGTCAGG	AAGCCACTCCTCCCACCATCAG
CeCNGC11	GAGGTGCTTGCTGGTACTTACTTGG	GAAGTTGTGGTGCCGTAGTAGATGG
CeCNGC12	GGAGCACCGCCCTGTCTTTATATG	CACGTAAGCCATCCTCATCTGTAGC
CeCNGC13	CGTCGGCACAAGAGAAGGAAGATG	TCGCTTGGCTCTCCGCTCTC
TUB	GCAGTTTACGGCGATGTTCA	ACTCTTCCTCGTCAGCTGTG

表2 CeCNGC家族成员的理化性质和亚细胞定位

Table 2 Physicochemical properties and subcellular localization of CeCNGC family members

基因名 Gene name	基因ID Gene ID	编码序列长度/bp CDS length	氨基酸数 Number of amino acids	分子量/D Molecular weight	等电点 pI
CeCNGC1	JL014344	1 461	488	55 806.43	8.23
CeCNGC2	JL001908	2 200	735	82 623.44	9.56
CeCNGC3	JL010106	1 029	343	40 046.06	9.14
CeCNGC4	JL001369	1 591	531	60 926.40	8.34
CeCNGC5	JL000073	2 105	703	81 211.91	8.94
CeCNGC6	JL006714	1 497	501	58 400.86	9.60
CeCNGC7	JL008860	1 869	625	71 627.58	9.35
CeCNGC8	JL007202	2 268	759	86 931.94	9.25
CeCNGC9	JL005182	879	293	34 538.52	9.81
CeCNGC10	JL002960	2 174	726	83 914.57	9.11
CeCNGC11	JL010726	1 953	653	74 573.27	9.70
CeCNGC12	JL019980	2 146	717	81 825.83	9.43
CeCNGC13	JL019514	1 313	438	50 481.31	8.94
基因名 Gene name	基因ID Gene ID	不稳定系数 Instability index	脂肪系数 Aliphatic index	总平均亲水系数 Grand average of hydropathicity	亚细胞定位 Subcellular localization
CeCNGC1	JL014344	41.29	80.39	-0.251	叶绿体Chloroplast
CeCNGC2	JL001908	58.92	92.49	0.019	质膜Plasma membrane
CeCNGC3	JL010106	54.80	79.91	-0.457	叶绿体Chloroplast
CeCNGC4	JL001369	43.95	90.19	-0.158	质膜Plasma membrane
CeCNGC5	JL000073	50.71	95.68	-0.103	质膜Plasma membrane
CeCNGC6	JL006714	50.76	92.26	-0.103	叶绿体Chloroplast
CeCNGC7	JL008860	50.00	96.74	0.012	质膜Plasma membrane
CeCNGC8	JL007202	42.34	89.80	-0.129	质膜Plasma membrane
CeCNGC9	JL005182	67.42	80.48	-0.616	细胞核nucleus
CeCNGC10	JL002960	54.21	87.34	-0.240	质膜Plasma membrane
CeCNGC11	JL010726	55.62	87.35	-0.270	质膜Plasma membrane
CeCNGC12	JL019980	56.53	93.05	0.041	质膜Plasma membrane
CeCNGC13	JL019514	53.37	76.64	-0.432	叶绿体Chloroplast

图1 建兰（Ce）、水稻（Os）和拟南芥（At）CNGC家族成员系统进化树

Fig. 1 Phylogenetic tree of CNGC family members in Cymbidium ensifolium（Ce），Oryza sativa（Os）and Arabidopsis thaliana（At）

图2 建兰基因组中CeCNGC染色体定位和基因共线性分析红线表示参与重复事件的CeCNGC基因对

Fig. 2 Chromosomal locations and segmental duplication of the CeCNGCs in the Cymbidium ensifolium genome The red lines inside indicate pairs of CeCNGCs involved in uplicated events

图3 建兰CeCNGC家族多序列比对（A）、基序组成和氨基酸序列（B、C）以及编码序列长度、非翻译区、外显子和内含子的统计分析（D）

Fig. 3 Multiple sequence alignment of the CeCNGC family in Cymbidium ensifolium（A）；motif composition and amino acid sequences（B，C）and CDS，UTR，exons，and introns（D）

图4 CeCNGC家族基因启动子区域顺式作用元件气泡图展示了CeCNGC基因家族内顺式作用元件的分布情况。横向折线图表示所有CeCNGC基因中每种顺式作用元件的累计数量，纵向折线图表示每个CNGC基因中顺式作用元件的总数

Fig. 4 Analysis of cis-acting elements in CeCNGCs promoters Bubble plot illustrates the distribution of cis-acting elements within the CeCNGC gene family. The horizontal line graph depicts the cumulative count of each cis-acting element across all CeCNGC，while the vertical line graph shows the total number of cis-acting elements present in each individual CeCNGC

图5 CeCNGC家族基因miRNA结合位点数量

Fig. 5 The number of miRNA binding sites in the CeCNGC gene family

图6 CeCNGC家族成员和miRNA调控网络

Fig. 6 The regulatory network between CeCNGC family members and miRNAs

图7 CeCNGC家族成员蛋白互作网络外圈表示与CeCNGC蛋白相互作用的蛋白质，内圈表示CeCNGC蛋白。颜色越深表示与其他成员的连接度越高。橙黄色圆圈表示仅涉及CeCNGC蛋白之间的相互作用网络

Fig. 7 Predicted protein-protein interaction networks of CeCNGC proteins The outer ring represents proteins interacting with CeCNGCs，while the inner ring represents the CeCNGC proteins themselves. The darker colors indicating higher connectivity with other members. The orange-yellow circles specifically illustrate the protein interaction network involving only CeCNGC proteins

图8 CeCNGC蛋白的磷酸化位点的数量（A）和激酶结合位点（B） unsp表示无特异性激酶预测结果

Fig. 8 The number of phosphorylation sites（A）and kinase binding sites（B）in CeCNGC proteins unsp indicates the prediction results of non-specific kinases

图9 CeCNGC家族成员表达模式热图（A）、不同植物组织基因表达趋势聚类分析（B）和不同花发育时期基因表达趋势聚类分析（C）加粗褐色折线表示该聚类基因的整体表达模式。L：叶；P：假鳞茎；R：根；B：苞片；Sep：萼片；Pet：花瓣；Lip：唇瓣；Gyn：合蕊柱；SB：小蕾；MB：中蕾；BB：大蕾；IF：初开期；BF：盛开期；SF：衰败期

Fig. 9 Heatmap（A），clustering analysis of gene expression trends in different plant tissues（B）clustering analysis of gene expression trends at different flower development stages（C）and of CeCNGC family members The thick brown line represents the overall expression pattern of the genes within the cluster. L：Leaf；P：Pseudobulb；R：Root；B：Bract；Sep：Sepal；Pet：Petal；Lip：Lip；Gyn：Gynandrium；SB：Samll bud；MB：Middle bud；BB：Big bud；IF：Initial bloom stage；BF：Blooming flowers；SF：Senescence stage

图10 CeCNGC1 ~ CeCNGC13在不同非生物胁迫下的RT-qPCR分析

Fig. 10 RT-qPCR analysis of CeCNGC1-CeCNGC13 under different abiotic stresses * P < 0.05，** P < 0.01

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建兰CNGC基因家族鉴定及其在非生物胁迫下的转录响应

Whole-Genome Identification of the CNGC Gene Family in Cymbidium ensifolium and Transcriptional Reponses Under Abiotic Stresses

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建兰CNGC基因家族鉴定及其在非生物胁迫下的转录响应

Whole-Genome Identification of the CNGC Gene Family in Cymbidium ensifolium and Transcriptional Reponses Under Abiotic Stresses

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参考文献 43

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