‘凤丹’牡丹PoSnRK基因家族鉴定及其在花衰老中的表达分析

doi:10.16420/j.issn.0513-353x.2024-0993

园艺学报 ›› 2025, Vol. 52 ›› Issue (9): 2363-2376.doi: 10.16420/j.issn.0513-353x.2024-0993

• 遗传育种·种质资源·分子生物学 • 上一篇下一篇

‘凤丹’牡丹PoSnRK基因家族鉴定及其在花衰老中的表达分析

王心茹¹^,^*, 白帅帅²^,^*, 孙雨乐¹, 武欣宇¹, 曾凤³, 刘春英¹, 张玉喜¹, 盖树鹏¹^,^**(), 袁延超¹^,^**()

¹ 青岛农业大学生命科学学院，山东青岛 266109
² 山东省农村经济管理服务总站，山东省农业农村厅，济南 250100
³ 齐河县农业农村局，山东德州 251199

收稿日期:2024-12-26 修回日期:2025-03-03 出版日期:2025-09-25 发布日期:2025-09-24
通讯作者:
^** E-mail：yanchao_y@126.com；
^** E-mail：yanchao_y@126.com；
作者简介:
^* 共同第一作者
基金资助:
国家自然科学基金项目(32201597); 国家自然科学基金项目(32271941); 青岛农业大学生命科学学院教学拔尖人才工程项目

Genome-Wide Identification of PoSnRK Gene Family in Paeonia ostii and Expression Analysis During Flower Senescence

WANG Xinru¹, BAI Shuaishuai², SUN Yule¹, WU Xinyu¹, ZENG Feng³, LIU Chunying¹, ZHANG Yuxi¹, GAI Shupeng¹^,^**(), YUAN Yanchao¹^,^**()

¹ College of Life Science， Qingdao Agricultural University，Qingdao， Shandong 266109， China
² Shandong Provincial Station of Rural Economic Management and Service， Shandong Provincial Department of Agriculture and Rural Affairs， Ji’nan 250100， China
³ Agricultural and Rural Bureau of Qihe County， Dezhou， Shandong 251199， China

Received:2024-12-26 Revised:2025-03-03 Published:2025-09-25 Online:2025-09-24

摘要/Abstract

摘要：

为了解牡丹（Paeonia suffruticosa）中蔗糖非发酵（SNF）1-相关蛋白激酶（sucrose non-fermenting-1-related protein kinase，SnRK）基因家族成员及其特征，利用生物信息的方法对该家族成员进行了鉴定，共鉴定出22个PoSnRK基因，包括3个SnRK1、6个SnRK2和13个SnRK3（CIPK）基因；保守结构域分析显示，所有PoSnRK1蛋白具有Pkinase、UBA和KA1结构域，PoSnRK2具有Pkinase结构域，PoCIPK具有Pkinase和NAF结构域；保守基序分析显示，Motif 15、Motif 17和Motif 18是SnRK1亚家族的特征基序，Motif 16和Motif 19是SnRK2的特征基序，Motif 7、Motif 10和Motif 14是CIPK的特征基序；共线性分析表明，PoSnRK基因间仅有1对共线性基因对Po04_CIPK10和Po01_CIPK14；顺势作用元件分析发现大部分PoSnRK启动子区域具有响应激素以及转录因子结合的顺式作用元件；转录表达谱及qRT-PCR结果显示，Po02_SnRK2.8、Po01_SnRK1.1、Po01_CIPK6和Po04_CIPK10的表达与牡丹花衰老过程相符，推测这四个基因在牡丹花衰老过程中发挥作用。

关键词: 牡丹, 蔗糖非发酵（SNF）1-相关蛋白激酶, 基因家族, 花, 衰老

Abstract:

To investigate the members and characteristics of sucrose non-fermenting-1-related protein kinase（SnRK）gene family in Paeonia suffruticosa，bioinformatics methods were employed to identify the members，resulting in the identification of 22 PoSnRKs，including 3 SnRK1s，6 SnRK2s，and 13 SnRK3s（CIPKs）；conserved domain analysis revealed that all PoSnRK1 proteins possessed the Pkinase，UBA，and KA1 domains，PoSnRK2s possessed the Pkinase domain，and PoCIPKs possessed the Pkinase and NAF domains；conserved motif analysis identified Motif 15，Motif 17，and Motif 18 as characteristic motifs of the SnRK1 subfamily，Motif 16 and Motif 19 as characteristic motifs of the SnRK2 subfamily，and Motif 7，Motif 10，and Motif 14 as characteristic motifs of the CIPK subfamily. Collinearity analysis indicated that there was only one pair of collinear PoSnRKs，Po04_CIPK10 and Po01_CIPK14；cis-acting element analysis found that most PoSnRK promoters contained cis-elements responsive to hormones and transcription factor binding. Transcriptome expression profiles and qRT-PCR results showed that the expression level of Po02_SnRK2.8，Po01_SnRK1.1，Po01_CIPK6 and Po04_CIPK10 was consistent with the flower senescence，suggesting that these four genes played a role in the flower aging process of tree peony.

Key words: tree peony, sucrose non-fermenting-1-related protein kinase, gene family, flowers, senescence

王心茹, 白帅帅, 孙雨乐, 武欣宇, 曾凤, 刘春英, 张玉喜, 盖树鹏, 袁延超. ‘凤丹’牡丹PoSnRK基因家族鉴定及其在花衰老中的表达分析[J]. 园艺学报, 2025, 52(9): 2363-2376.

WANG Xinru, BAI Shuaishuai, SUN Yule, WU Xinyu, ZENG Feng, LIU Chunying, ZHANG Yuxi, GAI Shupeng, YUAN Yanchao. Genome-Wide Identification of PoSnRK Gene Family in Paeonia ostii and Expression Analysis During Flower Senescence[J]. Acta Horticulturae Sinica, 2025, 52(9): 2363-2376.

图/表 10

表1 本研究中所用引物信息

Table 1 Primer information in this study

基因名称 Gene name	引物名称 Primer name	核苷酸序列（5′-3′） Nucleotide sequence
Po01_CIPK14	Po01_CIPK14-QF	GCCAGAGATCGGACATGGTT
Po01_CIPK14	Po01_CIPK14-QR	GCCAAATAAGGCGTTAGCCG
Po01_SnRK1.1	Po01_SnRK1.1-QF	TCATCTGGCAGCAGGTCTTC
Po01_SnRK1.1	Po01_SnRK1.1-QR	CACTTTACCAAATGCCCCGC
Po02_SnRK2.8	Po02_SnRK2.8-QF	TCCGAGTTTCCGAAGAGTGC
Po02_SnRK2.8	Po02_SnRK2.8-QR	CATCAGTGCTCTGGTGGCTT
Po04_CIPK10	Po04_CIPK10-QF	ACTGGCATGAGTGTGGCAAT
Po04_CIPK10	Po04_CIPK10-QR	CAGCTCACCACCCTTGACAT
Po01_CIPK6	Po01_CIPK6-QF	TGCAAGCAATCGGAGACACT
Po01_CIPK6	Po01_CIPK6-QR	TTCACCTCCTTGGCGACATC
PsActin	PsActin-QF	GTTCCAGCCATCACTAATCGG
PsActin	PsActin-QR	GAATAGACCCTCCAATCCAGACAC

图1 ‘凤丹’牡丹（Po）与拟南芥SnRK蛋白的系统发育树

Fig. 1 Phylogenetic tree of SnRK proteins in tree peony‘Fengdan’and Arabidopsis thaliana

图2 ‘凤丹’牡丹PoSnRK保守基序和基因结构

Fig. 2 The conserved motifs and gene structure of PoSnRK in tree peony‘Fengdan’

表2 ‘凤丹’牡丹PoSnRK蛋白的理化性质及亚细胞定位

Table 2 The physicochemical properties and subcellular location of PoSnRK in tree peony‘Fengdan’

转录本序号 Transcript ID	亚家族 Subfamily	氨基酸数 Number of amino acid	分子量/kD Molecular weight	等电点 pI	总平均亲水系数 Grand average of hydropathy		亚细胞定位 Subcellular location
Po01_SnRK1.1	SnRK1	571	65.12	9.29	-0.282		细胞膜Cell membrane
Po02_SnRK1.1	SnRK1	519	59.17	8.54	-0.211		质体Plastid
PoUn1_SnRK1.1	SnRK1	461	52.97	8.57	-0.086		细胞膜Cell membrane
Po04_SnRK2.3	SnRK2-III	363	41.13	4.89	-0.260		细胞核Nucleus
Po03_SnRK2.4	SnRK2-I	314	35.86	5.24	-0.607		细胞质Cytoplasm细胞核 Nucleus
Po03_SnRK2.6	SnRK2-III	363	41.39	4.92	-0.367		细胞核Nucleus
Po04_SnRK2.7	SnRK2-II	309	34.77	5.02	-0.264		细胞核Nucleus
Po02_SnRK2.8	SnRK2-II	297	33.76	5.76	-0.252		质体Plastid
Po04_SnRK2.8	SnRK2-II	331	37.85	5.51	-0.195		细胞膜Cell membrane
Po03_CIPK1	SnRK3	456	51.42	8.31	-0.366		细胞核Nucleus
Po02_CIPK3	SnRK3	491	56.23	7.54	-0.310		质体Plastid
Po03_CIPK4	SnRK3	445	48.81	8.79	-0.027		质体Plastid
Po03_CIPK5	SnRK3	648	72.37	8.67		-0.455		细胞核Nucleus
Po01_CIPK6	SnRK3	437	49.06	8.98	-0.331		细胞质Cytoplasm
Po05_CIPK6	SnRK3	428	48.25	9.35	-0.250		细胞质Cytoplasm
Po01_CIPK10	SnRK3	415	46.89	9.37	-0.290		细胞质Cytoplasm
Po04_CIPK10	SnRK3	531	60.87	8.79	-0.366		细胞核Nucleus
PoUn2035_CIPK10	SnRK3	422	47.95	9.35	-0.318		质体Plastid
PoUn267_CIPK10	SnRK3	447	50.60	8.98	-0.341		细胞膜Cell membrane
Po01_CIPK14	SnRK3	443	50.43	7.61		-0.313		质体Plastid
Po05_CIPK20	SnRK3	450	50.91	8.45	-0.426		细胞核Nucleus
Po02_CIPK21	SnRK3	469	52.62	5.77	-0.426		细胞质Cytoplasm 细胞核Nucleus

图4 ‘凤丹’牡丹（Paeonia ostii）SnRK基因与拟南芥（Arabidopsis thaliana）、葡萄（Vitis vinifera）、大花黄牡丹（Paeonia ludlowii）SnRK基因间的共线性分析

Fig. 4 The collinearity analysis between SnRK in Paeonia ostii and SnRK in Arabidopsis thaliana，Vitis vinifera，and Paeonia ludlowii

图3 ‘凤丹’牡丹PoSnRK在染色体上的分布（A）及基因间的共线性（B）

Fig. 3 The chromosomal location（A）and the collinearity analysis（B）of PoSnRK in tree peony‘Fengdan’

图5 PoSnRK启动子区域顺式作用元件 ABA：脱落酸响应元件；Aux：生长素响应元件；MeJA：茉莉酸响应元件；SA：水杨酸响应元件；GA：赤霉素响应元件；Wound：伤害响应原件；Dehydration：脱水响应元件；Drought：干旱响应元件；Low-temperature：低温响应元件；Anoxic：缺氧响应元件；Stress：胁迫响应元件；Defense and stress：防御和胁迫响应元件；Light：光响应元件；Anaerobic：厌氧诱导表达响应元件；Circadian：昼夜节律表达响应元件；Zein metabolism：玉米醇溶蛋白代谢调控响应元件；Flavonoid biosynthesi：类黄酮生物合成响应元件；Root：根中特异表达响应元件；Seed：种子中特异表达响应元件；Endosperm：胚乳中特异表达响应元件；Endosperm-nagetive：胚乳中特异负向表达响应元件；Meristem：分生组织表达相关响应元件；AP-1：转录因子AP-1响应元件；MYB：转录因子MYB响应元件；MYC：转录因子MYC响应元件；Phytohormone：激素相关响应元件；Stress：胁迫相关响应元件；Progress：生物过程相关响应元件；Tissue：组织特异表达相关响应元件；Transcription factor：转录因子响应元件；左图中数字代表顺式作用元件的个数，右图代表各类别的顺式作用元件的总数

Fig. 5 cis-Acting elements in the promoter of PoSnRK ABA：Abscisic acid response element；Aux：Auxin response element；MeJA：Jasmonic acid response element；SA：Salicylic acid response element； GA：Gibberellin response element；Wound：Wound response element；Dehydration：Dehydration response element；Drought：Drought response element；Low-temperature：Low-temperature response element；Anoxic：Anoxic response element；Stress：Stress response element；Defense and stress：Defense and stress response element；Light：Light response element；Anaerobic：Anaerobic induction expression response element；Circadian：Circadian expression response element；Zein metabolism：Zein metabolism response element；Flavonoid biosynthesis：Flavonoid biosynthesis response element；Root：Root-specific expression response element；Seed：Seed-specific expression response element；Endosperm：Endosperm-specific expression response element；Endosperm-negative：Endosperm-specific negative expression response element；Meristem：Meristem expression-related response element；AP-1：Transcription factor AP-1 response element；MYB：Transcription factor MYB response element；MYC：Transcription factor MYC response element； Phytohormone：Phytohormone-related response element；Stress：Stress-related response element；Progress：Biological process-related response element；Tissue：Tissue-specific expression-related response element；Transcription factor：Transcription factor response element；In left panel，the numbers in the grid represent the number of cis-elements，and in right panel，the bar chart represents the number of cis-elements，of each category

图6 PoSnRK在牡丹‘雪映桃花’花衰老过程中表达谱分析 S1：初开期；S2：半开期；S3：盛开期；S4：初衰期；S5：衰老期。下同

Fig. 6 Expression profile analysis of PoSnRK during flower senescence in peony‘Xueying Taohua’ S1：Initial blooming stage；S2：Half-opening stage；S3：Full blooming stage；S4：Initial wilting stage；S5：Aging stage. The same below

图7 PoSnRK在牡丹花衰老过程中相对表达水平不同小写字母表示在不同发育时期差异显著，P < 0.05

Fig. 7 The relative expression level of PoSnRK during peony flower senescence Different lowercase letters indicate significant differences at different developmental stages，P < 0.05

图8 PoSnRK启动子结合转录因子预测

Fig. 8 Prediction of transcription factors binding to PoSnRK promoters

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‘凤丹’牡丹PoSnRK基因家族鉴定及其在花衰老中的表达分析

Genome-Wide Identification of PoSnRK Gene Family in Paeonia ostii and Expression Analysis During Flower Senescence

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‘凤丹’牡丹PoSnRK基因家族鉴定及其在花衰老中的表达分析

Genome-Wide Identification of PoSnRK Gene Family in Paeonia ostii and Expression Analysis During Flower Senescence

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