甘蔗属割手密与热带种PEBP基因家族的鉴定及其开花调控功能分析

doi:10.3864/j.issn.0578-1752.2026.04.003

中国农业科学 ›› 2026, Vol. 59 ›› Issue (4): 734-749.doi: 10.3864/j.issn.0578-1752.2026.04.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

甘蔗属割手密与热带种PEBP基因家族的鉴定及其开花调控功能分析

罗正英¹^,³(), 胡嗣桢¹(), 林秀琴²^,³, 胡鑫²^,³, 张敏³, 徐超华²^,³, 刘新龙²^,³(), 曾千春¹^,⁴()

¹ 云南农业大学农学与生物技术学院/云南省作物生产与智慧农业重点实验室，昆明 650201
² 云南省农业科学院热带作物生物育种全国重点实验室，昆明 650205
³ 云南省农业科学院甘蔗研究所/云南甘蔗遗传改良重点实验室/农业农村部甘蔗生物学与遗传育种重点实验室（云南），云南开远 661699
⁴ 云南农业大学热带作物学院，云南普洱 665099

收稿日期:2025-08-10 出版日期:2026-02-10 发布日期:2026-02-10
通信作者:
刘新龙，E-mail：liuxinlong@yaas.org.cn。
曾千春，E-mail：zqch@yahoo.com.cn
联系方式: 罗正英，E-mail：zhengyluo@163.com。胡嗣桢，E-mail：18973644103@163.com。罗正英和胡嗣桢为同等贡献作者。
基金资助:
国家重点研发计划(2022YFD2301101); 云南种子种业联合实验室(202205AR070001-09); 云南省农业科学院科研项目(2024KYZX-03)

Identification and Functional Characterization of the PEBP Gene Family in Regulating Flowering Time in Saccharum spontaneum and Saccharum officinarum

LUO ZhengYing¹^,³(), HU SiZhen¹(), LIN XiuQin²^,³, HU Xin²^,³, ZHANG Min³, XU ChaoHua²^,³, LIU XinLong²^,³(), ZENG QianChun¹^,⁴()

¹ College of Agronomy and Biotechnology, Yunnan Agricultural University/The Key Laboratory for Crop Production and Smart Agriculture of Yunnan Province, Kunming 650201
² State Key Laboratory of Tropical Crop Breeding, Yunnan Academy of Agricultural Sciences, Kunming, 650205
³ Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences/Yunnan Key Laboratory of Sugarcane Genetic Improvement/Key Laboratory of Sugarcane Biology and Genetic Breeding (Yunnan), Ministry of Agriculture and Rural Affairs Kaiyuan 661699, Yunnan
⁴ College of Tropical Crops, Yunnan Agricultural University, Puer 665099, Yunnan

Received:2025-08-10 Published:2026-02-10 Online:2026-02-10

摘要/Abstract

摘要：

【目的】磷脂酰乙醇胺结合蛋白（phosphatidyl ethanolamine-binding protein，PEBP）是调控植物开花的关键因子，通过协调光周期、植物激素等信号精确调控开花时间。以甘蔗原始亲本割手密（Saccharum spontaneum）和热带种（Saccharum officinarum）为材料，系统鉴定PEBP基因家族成员，分析其基本结构、进化特征与表达模式，验证关键基因功能，为解析甘蔗开花调控机制及分子育种提供理论依据。【方法】基于拟南芥和水稻PEBP基因家族成员的蛋白序列，对割手密和热带种基因组数据进行比对分析，并结合PEBP基因对应的隐马尔可夫模型文件PF01161进行筛选，最终鉴定PEBP基因家族成员；利用MEGA-X、TBtools等软件进行系统进化、基因结构、保守基序和共线性分析；通过PlantCARE数据库预测启动子顺式作用元件；结合转录组数据和qRT-PCR结果分析基因表达模式；构建FT3和TFL1过表达载体并转化拟南芥，从而验证其功能。【结果】割手密和热带种分别含有23和20个PEBP基因，均分为FT-like、TFL1-like和MFT-like 3个亚家族。蛋白理化性状分析表明，割手密PEBP蛋白（151—364 aa，17.1—40.4 kDa）的变异范围显著大于热带种（170—191 aa，19.2—20.8 kDa），但均以亲水性碱性蛋白为主。保守基序分析揭示，TFL1-like亚家族高度保守，MFT-like则呈现显著多样性，其中，割手密SspMFT1.1/1.2/1.3具有独特的结构域延长。基因结构显示，FT-like外显子数目变异最大（2—5个），高于TFL1-like（3—4个）和MFT-like（4个）。进化研究发现，PEBP基因主要通过全基因组复制和片段复制扩张，热带种串联复制频率（8%）显著高于割手密（1%）。共线性分析表明，部分基因对（如SspFT5.1/SspFT5.2和SoFT12.1/SoFT12.2）显示出物种特异性复制特征，割手密PEBP基因家族成员与高粱保守性更高。启动子分析显示，割手密和热带种PEBP基因均富含光响应和激素响应元件，其中，茉莉酸和脱落酸响应元件占比最高。表达分析鉴定出FT3和TFL1 2个关键基因，前者表达持续上调且光周期敏感，后者呈先降后升趋势，且不受光周期调控。转基因分析证实SspFT3和SoFT3促进拟南芥开花（提前约19 d），而SspTFL1和SoTFL1则抑制开花。【结论】在割手密和热带种中分别鉴定了23和20个PEBP基因，分为FT-like、TFL1-like和MFT-like 3个亚家族，各亚家族在基因结构和蛋白特性方面呈现明显分化。FT3和TFL1在开花进程中呈现相反的表达模式，其中，FT3对光周期响应敏感，且在热带种中表达量显著低于割手密。FT3具有促进开花的典型功能，而TFL1抑制开花。

关键词: 割手密, 热带种, PEBP基因家族, 开花调控, 表达分析, 光周期

Abstract:

【Objective】The phosphatidylethanolamine-binding protein (PEBP) family serves as pivotal regulators of plant flowering, orchestrating floral transition through the coordination of photoperiodic and hormonal signaling pathways. This study systematically characterized PEBP gene family members in the ancestral Saccharum species S. spontaneum and S. officinarum, elucidating their structural architecture, evolutionary trajectories, and expression profiles to unravel the molecular mechanisms governing flowering regulation and facilitate molecular breeding strategies.【Method】The PEBP gene family was identified by performing sequence alignment of Arabidopsis and rice PEBP protein sequences against the genomes of S. spontaneum and S. officinarum, supplemented with screening using the hidden Markov model profile PF01161. Phylogenetic reconstruction, gene structure analysis, conserved motif identification, and synteny evaluation were performed using MEGA-X and TBtools. Putative cis-regulatory elements within promoter regions were predicted via the PlantCARE database. Transcriptomic profiling coupled with qRT-PCR validation delineated expression dynamics, while functional characterization of FT3 and TFL1 genes was achieved through Arabidopsis transformation.【Result】Our analysis identified 23 and 20 PEBP genes in S. spontaneum and S. officinarum, respectively, classified into three subfamilies: FT-like, TFL1-like, and MFT-like. Protein characterization revealed greater variability in S. spontaneum PEBPs (151-364 aa; 17.1-40.4 kDa) compared to S. officinarum (170-191 aa; 19.2-20.8 kDa), though both predominantly encoded hydrophilic alkaline proteins. Conserved motif analysis demonstrated stringent conservation in TFL1-like subfamily members, while MFT-like proteins exhibited remarkable structural plasticity, exemplified by elongated PEBP domains in SspMFT1.1/1.2/1.3. Gene architecture analysis showed FT-like members possessed the highest exon variability (2-5), contrasting with TFL1-like (3-4) and MFT-like (fixed 4-exon) subfamilies. Evolutionary analysis revealed whole-genome and segmental duplications as primary expansion mechanisms, with tandem duplication frequency substantially higher in S. officinarum (8%) than S. spontaneum (1%). Syntenic analysis uncovered species-specific duplication events (e.g., SspFT5.1/SspFT5.2 and SoFT12.1/SoFT12.2) and stronger conservation between S. spontaneum and sorghum. Promoter analysis identified abundant light- and hormone-responsive elements, particularly jasmonic acid and abscisic acid response motifs. Expression profiling identified two key genes, FT3 and TFL1, with contrasting expression dynamics. FT3 displayed a sustained, photoperiod-sensitive upregulation, whereas TFL1 showed an initial decrease followed by an increase and was independent of photoperiod regulation. Transgenic validation confirmed FT3 orthologs consistently accelerated flowering (~19 days early), whereas TFL1 genes functioned as floral repressors.【Conclusion】Genomic analysis revealed 23 and 20 PEBP genes in S. spontaneum and S. officinarum, respectively, phylogenetically clustered into three well-defined subfamilies (FT-like, TFL1-like, and MFT-like) that exhibited substantial structural and functional divergence. FT3 and TFL1 demonstrated opposite expression patterns during floral induction, with FT3 showing marked photoperiod responsiveness and significantly elevated transcript abundance in S. spontaneum relative to S. officinarum. Transgenic functional analysis confirmed that FT3 typically promotes flowering, whereas the TFL1 gene suppresses it.

Key words: Saccharum spontaneum, Saccharum officinarum, PEBP gene family, flowering regulation, expression profiling, photoperiod

罗正英, 胡嗣桢, 林秀琴, 胡鑫, 张敏, 徐超华, 刘新龙, 曾千春. 甘蔗属割手密与热带种PEBP基因家族的鉴定及其开花调控功能分析[J]. 中国农业科学, 2026, 59(4): 734-749.

LUO ZhengYing, HU SiZhen, LIN XiuQin, HU Xin, ZHANG Min, XU ChaoHua, LIU XinLong, ZENG QianChun. Identification and Functional Characterization of the PEBP Gene Family in Regulating Flowering Time in Saccharum spontaneum and Saccharum officinarum[J]. Scientia Agricultura Sinica, 2026, 59(4): 734-749.

图/表 8

图1

表1

割手密和热带种PEBP基因家族成员的理化特征"

物种 Species	蛋白 Protein	染色体 Chromosome	亚族分类 Subfamily classification	氨基酸数 No. of amino acids (aa)	分子量Molecular weight (kDa)	等电点 pI	不稳定系数Instability index	脂肪族指数Aliphatic index	平均亲水指数Grand average of hydropathicity
割手密 S. spontaneum	SspFT1.1	Chr.6C	FT-Like	173	19.365	7.94	34.57	76.01	-0.236
	SspFT1.2	Chr.10A	FT-Like	191	21.227	5.57	40.07	78.48	-0.206
	SspFT2	Chr.3D	FT-Like	191	21.040	8.47	49.68	75.45	-0.285
	SspFT3	Chr.3D	FT-Like	173	19.299	6.73	42.61	85.55	-0.131
	SspFT4	Chr.10A	FT-Like	179	19.978	7.73	30.58	75.53	-0.244
	SspFT5.2	Chr.5B	FT-Like	364	40.368	10.08	51.96	78.52	-0.163
	SspFT5.1	Chr.9B	FT-Like	180	20.355	8.76	41.91	64.39	-0.451
	SspFT6	Chr.6C	FT-Like	151	17.101	9.33	31.35	81.26	-0.307
	SspFT7	Chr.4C	FT-Like	185	20.517	6.90	40.35	69.51	-0.301
	SspFT8	Chr.3D	FT-Like	177	20.089	9.09	40.34	88.02	-0.132
	SspFT9	Chr.4C	FT-Like	174	19.676	8.56	42.54	77.18	-0.389
	SspFT10	Chr.9B	FT-Like	175	19.880	7.89	40.52	79.09	-0.264
	SspFT11	Chr.8B	FT-Like	176	19.249	6.89	40.75	74.83	-0.228
	SspFT12	Chr.2D	FT-Like	178	19.987	7.85	43.09	84.89	-0.303
	SspFT13	Chr.6C	FT-Like	185	20.625	6.41	42.94	68.54	-0.305
	SspTFL1	Chr.5B	TFL-Like	173	19.595	7.96	48.39	86.65	-0.150
	SspTFL2	Chr.1D	TFL-Like	197	22.540	9.77	52.12	73.15	-0.467
	SspTFL3	Chr.4C	TFL-Like	173	19.517	8.76	37.76	78.21	-0.294
	SspTFL4	Chr.8B	TFL-Like	173	19.532	8.82	44.44	83.24	-0.258
	SspMFT1.1	Chr.10A	MFT-Like	302	33.077	10.01	55.20	84.04	-0.064
	SspMFT1.2	Chr.10B	MFT-Like	268	29.412	10.35	48.32	88.84	-0.025
	SspMFT1.3	Chr.10C	MFT-Like	273	30.119	10.54	43.96	87.25	-0.094
	SspMFT2	Chr.3D	MFT-Like	171	18.718	8.62	35.35	85.50	-0.139
热带种 S. officinarum	SoFT1	Chr.10A	FT-Like	175	19.651	6.91	38.87	74.00	-0.305
	SoFT2	Chr.02H	FT-Like	191	20.848	8.59	53.76	79.53	-0.242
	SoFT3	Chr.03B	FT-Like	173	19.341	6.73	42.61	87.23	-0.116
	SoFT4	Chr.10A	FT-Like	179	19.978	7.73	30.58	75.53	-0.244
	SoFT5.1	Chr.05D	FT-Like	177	20.061	8.64	44.88	69.32	-0.407
	SoFT5.2	Chr.09G	FT-Like	176	19.869	9.00	46.31	69.72	-0.404
	SoFT6	Chr.06H	FT-Like	174	19.627	7.72	33.87	77.76	-0.386
	SoFT7	Chr.04C	FT-Like	185	20.517	6.90	40.35	69.51	-0.301
	SoFT8	Chr.03B	FT-Like	177	20.005	9.38	39.33	85.25	-0.156
	SoFT9	Chr.04C	FT-Like	174	19.676	8.56	42.54	77.18	-0.389
	SoFT10	Chr.09B	FT-Like	175	19.876	7.89	40.52	79.09	-0.269
	SoFT11	Chr.08E	FT-Like	176	19.176	7.71	46.44	75.97	-0.186
	SoFT12.2	Chr.02H	FT-Like	178	19.979	7.87	42.45	82.70	-0.325
	SoFT12.1	Chr.02C	FT-Like	170	19.029	8.82	43.18	84.29	-0.279
	SoFT13	Chr.06H	FT-Like	185	20534	6.50	38.75	68.54	-0.282
	SoTFL1	Chr.05D	TFL-Like	173	19.556	7.94	50.38	86.07	-0.146
	SoTFL2	Chr.06H	TFL-Like	173	19.669	9.25	44.16	75.95	-0.299
	SoTFL3	Chr.04C	TFL-Like	173	19.517	8.76	37.76	78.21	-0.294
	SoTFL4	Chr.08E	TFL-Like	173	19.533	7.93	45.21	83.24	-0.258
	SoMFT1	Chr.10A	MFT-Like	181	19.630	6.96	42.23	87.29	0.024

表1

图2

图3

图4

表2

图5

图6

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粤糖93-159基因编号 Yuetang 93-159 gene ID	热带种比对 Homology to S. officinarum		割手密比对 Homology to S. spontaneum		早期花穗vs 茎尖分生组织 Early panicle_vs_SAM		中期花穗vs 茎尖分生组织 Mid panicle_vs_SAM		晚期花穗vs 茎尖分生组织 Late panicle_vs_SAM
粤糖93-159基因编号 Yuetang 93-159 gene ID	同源基因Ortholog	一致性 Identity (%)	同源基因Ortholog	一致性 Identity (%)	Log₂FC	P值 P value	Log₂FC	P值 P value	Log₂FC	P值 P value
TRINITY_DN783_c1_g1	SoFT3	99.62	SspFT3	99.62	4.4855	0.0205	6.1656	0.0004	6.2672	0.0004
TRINITY_DN65785_c0_g1_orf1	SoTFL1	99.09	SspTFL1	98.63	-0.9004	0.0562	-0.6374	0.0688	1.0995	0.0233

甘蔗属割手密与热带种PEBP基因家族的鉴定及其开花调控功能分析

Identification and Functional Characterization of the PEBP Gene Family in Regulating Flowering Time in Saccharum spontaneum and Saccharum officinarum

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