白桦BpLUXs基因鉴定与功能验证

doi:10.7525/j.issn.1673-5102.2026.03.003

摘要/Abstract

摘要：

光周期（photoperiod）是调控林木季节性生长与生长停止的关键环境因子。在光周期信号感知和转导过程中，生物钟晚间复合体（evening complex，EC）的核心组件-节律失常蛋白（LUX ARRHYTHMO，LUX）在其中发挥重要作用。本研究在白桦（Betula platyphylla）参考基因组中分析得到47个BpLUXs同源基因。生物信息学分析表明，BpLUX1、BpLUX5和BpLUX18具有典型的Evening Element（EE）顺式作用元件，BpLUX1、BpLUX5和BpLUX18均具有典型的MYB保守结构域，BpLUX1、BpLUX5具有典型的昼夜节律表达模式。通过CRISPR/Cas9技术靶向敲除BpLUX1和BpLUX5，成功创制BpLUX1功能缺失突变体bplux1（BpLUX5基因敲除未成功）。表型分析结果表明，在短日照（short-day，SD）条件下，野生型白桦响应短光周期在第19天开始几乎全部停止生长，而bplux1突变体仍保持活跃生长状态，同时，野生型白桦株高未表现出显著性差异，而bplux1突变体株高显著增高，表现出延迟生长停止表型，表明BpLUX1是白桦响应短日照、诱导生长停止的关键基因。转录组分析进一步揭示，在SD条件下，bplux1突变体中脱落酸（abscisic acid，ABA）信号通路中脱落酸受体调控组分1基因BpRCAR1（Regulatory Component of ABA Receptor 1）等11个差异表达基因下调，赤霉素（gibberellin，GA）信号通路中赤霉素不敏感类似物1基因BpRGL1（RGA-Like 1）等7个差异表达基因上调，并且BpLUX1与ABA和GA信号通路中的关键差异表达基因有潜在的结合位点。这表明BpLUX1基因可能通过影响植物激素信号进一步影响白桦生长停止。综上所述，本研究为解析BpLUX1在光周期调控白桦生长停止的过程中发挥功能提供了一定的理论依据。

关键词: 白桦, BpLUX, 光周期, 生长停止

Abstract:

Photoperiod serves as the key environmental cue governing seasonal growth and growth cessation in forest trees. Within the perception and transduction of photoperiodic signals， LUX ARRHYTHMO（LUX）， a core component of the circadian clock Evening Complex（EC）， plays a pivotal role. In the present study， a total of 47 BpLUX homologous genes were identified through genome-wide analysis in the reference genome of Betula platyphylla. Bioinformatic analyses revealed that BpLUX1， BpLUX5， and BpLUX18 harbor typical Evening Element（EE） cis-acting elements. All three genes possessed characteristic MYB conserved domains， while BpLUX1 and BpLUX5 additionally exhibited typical circadian rhythmic expression patterns. Using CRISPR/Cas9-mediated genome editing， loss-of-function mutants for BpLUX1（bplux1） were successfully generated， whereas targeted knockout of BpLUX5 proved unsuccessful. Phenotypic characterization demonstrated that under short-day（SD） conditions， wild-type birch ceased growth almost entirely by day 19 in response to shortened photoperiods， whereas bplux1 mutants remained actively growing. Furthermore， while no significant difference in plant height was observed in wild-type plants， bplux1 mutants displayed significantly increased height， exhibiting a delayed growth cessation phenotype. These findings indicate that BpLUX1 is a key gene mediating short-day responses and growth cessation in birch. Transcriptome analysis further revealed that under SD conditions， eleven differentially expressed genes（DEGs） were down-regulated in bplux1 mutants， including BpRCAR1 （Regulatory Component of ABA Receptor 1） involved in ABA （abscisic acid） signaling， whereas seven DEGs were up-regulated， including BpRGL1（RGA-Like1） associated with GA （gibberellin） signaling. Notably， potential binding sites between BpLUX1 and key DEGs in ABA and GA signaling pathways were identified， suggesting that BpLUX1 may influence growth cessation through modulation of phytohormone signaling. Collectively， this study provides a theoretical basis for understanding the role of BpLUX1 in photoperiod-regulated growth cessation in birch.

Key words: Betula platyphylla, BpLUX, photoperiod, growth cessation

中图分类号:

S714.7

潘梦婷, 刘月影, 郑志民. 白桦BpLUXs基因鉴定与功能验证[J]. 植物研究, 2026, 46(3): 411-424.

Mengting PAN, Yueying LIU, Zhimin ZHENG. Identification and Preliminary Functional Analysis of the BpLUXs Gene in Betula platyphylla Suk.[J]. Bulletin of Botanical Research, 2026, 46(3): 411-424.

图/表 8

表1

表2

表3

BpLUXs 基因比对结果

基因名称 Gene name	基因号 Gene ID	比对率 Identity/%	比对长度 Alignment length/bp	氨基酸数目 Number of amino acids/aa	等电位点 Theoretical pI	相对分子质量 Relative molecular mass/Da	亚细胞定位 Subcellular location
BpLUX1	Bplat_h1_10G00567	54.472	246	301	5.47	32 910.07	细胞核Nucleus
BpLUX2	Bplat_h1_14G01337	61.789	123	321	6.19	34 563.60	细胞核Nucleus
BpLUX3	Bplat_h1_08G02011	43.966	116	670	6.08	71 886.79	细胞核Nucleus
BpLUX4	Bplat_h1_08G01216	56.962	79	680	5.94	74 370.65	细胞核Nucleus
BpLUX5	Bplat_h1_01G00318	63.235	68	694	5.97	76 202.68	细胞核Nucleus
BpLUX6	Bplat_h1_12G02326	62.319	69	714	5.38	78 100.19	细胞核Nucleus
BpLUX7	Bplat_h1_11G01513	32.911	158	737	5.18	81 496.51	细胞核Nucleus
BpLUX8	Bplat_h1_06G00273	58.730	63	483	5.10	48 822.07	细胞核Nucleus
BpLUX9	Bplat_h1_08G01998	46.067	89	188	5.51	20 938.59	细胞核Nucleus
BpLUX10	Bplat_h1_06G00274	58.730	63	1051	5.81	118 236.64	细胞核Nucleus
BpLUX11	Bplat_h1_01G01163	37.671	146	605	5.67	67 851.41	细胞核Nucleus
BpLUX12	Bplat_h1_05G00214	48.837	86	456	7.98	52 252.68	细胞核Nucleus
BpLUX13	Bplat_h1_11G02231	40.426	94	427	6.44	46 880.03	细胞核Nucleus
BpLUX14	Bplat_h1_02G00164	53.333	60	560	6.08	62 528.25	细胞核Nucleus
BpLUX15	Bplat_h1_06G01349	56.897	58	501	6.00	54 793.90	细胞核Nucleus
BpLUX16	Bplat_h1_14G02337	53.623	69	419	7.29	46 884.77	细胞核Nucleus
BpLUX17	Bplat_h1_04G00221	54.286	70	355	8.25	39 420.90	细胞核Nucleus
BpLUX18	Bplat_h1_07G02471	60.000	55	322	7.71	35 163.07	细胞核Nucleus
BpLUX19	Bplat_h1_08G03288	54.545	55	351	6.81	38 983.83	细胞核Nucleus
BpLUX20	Bplat_h1_03G01346	43.023	86	463	7.12	50 724.45	细胞核Nucleus
BpLUX21	Bplat_h1_08G00772	37.963	108	468	5.35	53 375.19	细胞核Nucleus
BpLUX22	Bplat_h1_14G00212	37.209	129	349	7.73	38 959.20	细胞核Nucleus
BpLUX23	Bplat_h1_06G00533	45.455	66	323	5.72	35 639.67	细胞核Nucleus
BpLUX24	Bplat_h1_09G01482	51.786	56	395	6.59	44 367.63	细胞核Nucleus
BpLUX25	Bplat_h1_12G00064	44.118	68	389	6.48	42 308.72	细胞核Nucleus
BpLUX26	Bplat_h1_03G00243	55.556	54	484	5.51	53 293.29	细胞核Nucleus
BpLUX27	Bplat_h1_02G00362	50.746	67	280	6.98	31 521.49	细胞核Nucleus
BpLUX28	Bplat_h1_08G01864	54.545	55	309	8.07	34 789.69	细胞核Nucleus
BpLUX29	Bplat_h1_06G02707	38.281	128	350	6.90	39 394.52	细胞核Nucleus
BpLUX30	Bplat_h1_07G01375	43.478	92	430	6.04	48 967.34	细胞核Nucleus
BpLUX31	Bplat_h1_07G02472	58.491	53	335	6.57	36 684.08	细胞核Nucleus
BpLUX32	Bplat_h1_07G02207	36.634	101	302	9.31	34 359.32	细胞核Nucleus
BpLUX33	Bplat_h1_05G01228	48.718	78	460	5.59	50 655.07	细胞核Nucleus
BpLUX34	Bplat_h1_10G00179	54.717	53	366	8.70	41 561.47	细胞核Nucleus
BpLUX35	Bplat_h1_14G01211	52.727	55	295	6.34	31 664.44	细胞核Nucleus
BpLUX36	Bplat_h1_12G00245	38.542	96	296	8.75	33 023.06	细胞核Nucleus
BpLUX37	Bplat_h1_14G01650	49.091	55	425	7.09	47 626.14	细胞核Nucleus
BpLUX38	Bplat_h1_12G01893	35.484	93	193	10.18	21 718.78	细胞核Nucleus
BpLUX39	Bplat_h1_12G01896	46.667	60	345	8.63	38 971.68	细胞核Nucleus
BpLUX40	Bplat_h1_02G00314	45.000	60	370	9.30	41 774.52	细胞核Nucleus
BpLUX41	Bplat_h1_11G00872	45.000	60	419	7.86	46 573.83	细胞核Nucleus
BpLUX42	Bplat_h1_11G01688	54.717	53	229	7.87	26 381.96	细胞核Nucleus
BpLUX43	Bplat_h1_14G01651	51.852	54	319	8.96	37 063.91	细胞核Nucleus
BpLUX44	Bplat_h1_14G01652	44.595	74	288	7.17	33 126.49	细胞核Nucleus
BpLUX45	Bplat_h1_14G00254	50.943	53	258	7.71	29 272.00	细胞核Nucleus
BpLUX46	Bplat_h1_13G02359	40.244	82	342	9.18	38 879.09	细胞核Nucleus
BpLUX47	Bplat_h1_07G02426	60.606	33	149	9.15	15 969.02	细胞核Nucleus

表3

图1

图2

图3

图4

图5

参考文献 49

[1]	关文彬.中国东北地区白桦林植被生态学的研究：桦属植物与中国白桦林的地理分布［J］.北京林业大学学报，1998，20（4）：104-109.
	GUAN W B.Vegetation ecology on communities dominated by Betula platyphylla in northeast of China distribution of communities dominated by Betula platyphylla ［J］.Journal of Beijing Forestry University，1998，20（4）：104-109.
[2]	IMAIZUMI T， KAY S A.Photoperiodic control of flowering：not only by coincidence［J］.Trends in Plant Science，2006，11（11）：550-558.
[3]	王飞飞.大豆生育期基因TOF7的图位克隆和功能分析［D］.哈尔滨：中国科学院大学（中国科学院东北地理与农业生态研究所），2019：2-12.
	WANG F F.Map-based cloning and functional analysis of the maturity gene TOF7 in soybean［D］.Harbin：Northeast Institute of Geography and Agroecology，Chinese Academy of Sciences，2019：2-12.
[4]	陈政.EF11调控大豆光周期开花的分子机制研究［D］.广州：广州大学，2025.
	CHEN Z.The molecular mechanism of EF11 in regulating photoperiod flowering in soybean［D］.Guangzhou：Guangzhou University，2025.
[5]	GARNER W W， ALLARD H A.Photoperiodism，the response of the plant to relative length of day and night［J］.Science，1922，55（1431）：582-583.
[6]	ANDRÉS F， COUPLAND G.The genetic basis of flowering responses to seasonal cues［J］.Annual Review of Plant Biology，2012，63：627-650.
[7]	WANG Z Y， TOBIN E M.Constitutive expression of the circadian clock associated 1（cca1） gene disrupts circadian rhythms and suppresses its own expression［J］.Cell，1998，93（7）：1207-1217.
[8]	MATSUSHIKA A， MAKINO S， KOJIMA M，et al.The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana：II.Characterization with CCA1-overexpressing plants［J］.Plant & Cell Physiology，2002，43（1）：118-122.
[9]	GENDRON J M， PRUNEDA-PAZ J L， DOHERTY C J，et al. Arabidopsis circadian clock protein，TOC1，is a DNA-binding transcription factor［J］.Proceedings of the National Academy of Sciences of the United States of America，2012，109（8）：3167-3172.
[10]	NAKAMICHI N， KIBA T， HENRIQUES R，et al.PSEUDO-RESPONSE REGULATORS 9，7，and 5 are transcriptional repressors in the Arabidopsis circadian clock［J］.The Plant Cell，2010，22（3）：594-605.
[11]	NUSINOW D A， HELFER A， HAMILTON E E，et al.The ELF4-ELF3-LUX complex links the circadian clock to diurnal control of hypocotyl growth［J］.Nature，2011，475（7356）：398-402.
[12]	DOYLE M R， DAVIS S J， BASTOW R M，et al.The ELF4 gene controls circadian rhythms and flowering time in Arabidopsis thaliana ［J］.Nature，2002，419（6902）：74-77.
[13]	HUANG W， PÉREZ-GARCÍA P， POKHILKO A，et al.Mapping the core of the Arabidopsis circadian clock defines the network structure of the oscillator［J］.Science，2012，336（6077）：75-79.
[14]	NASCIMENTO A S M DO， HENSCHEL J M， Felipe S H S，et al.Photoperiod and circadian regulation in plants：a review of insights from in vitro studies［J］.Biology，2025，14（11）：1502.
[15]	ZHANG C， GAO M， SEITZ N C，et al.LUX ARRHYTHMO mediates crosstalk between the circadian clock and defense in Arabidopsis ［J］.Nature Communications，2019，10：2543.
[16]	JONES M A， MOROHASHI K， GROTEWOLD E，et al. Arabidopsis JMJD5/JMJ30 acts independently of LUX ARRHYTHMO within the plant circadian clock to enable temperature compensation［J］.Frontiers in Plant Science，2019，10：57.
[17]	CHOW B Y， SANCHEZ S E， BRETON G，et al.Transcriptional regulation of LUX by CBF1 mediates cold input to the circadian clock in Arabidopsis ［J］.Current Biology，2014，24（13）：1518-1524.
[18]	WANG Y L， TONG W， LI F D，et al.LUX ARRHYTHMO links CBF pathway and jasmonic acid metabolism to regulate cold tolerance of tea plants［J］.Plant Physiology，2024，196（2）：961-978.
[19]	KIM H， KIM H J， VU Q T，et al.Circadian control of ORE1 by PRR9 positively regulates leaf senescence in Arabidopsis ［J］.Proceedings of the National Academy of Sciences of the United States of America，2018，115（33）：8448-8453.
[20]	XU P， ZHANG Y X， WEN X X，et al.The clock component OsLUX regulates rice heading through recruiting OsELF3-1 and OsELF4s to repress Hd1 and Ghd7 ［J］.Journal of Advanced Research，2023，48：17-31.
[21]	徐鹏.水稻生物钟基因OsLUX调控抽穗期的分子机制研究［D］.重庆：西南大学，2022.
	Xu P.Molecular mechanism research of rice biological clock gene OsLUX regulates heading date［D］.Chongqing：Southwest University，2022.
[22]	CAI Z Z， ZHANG Y D， TANG W Q，et al.LUX ARRHYTHMO interacts with ELF3a and ELF4a to coordinate vegetative growth and photoperiodic flowering in rice［J］.Frontiers in Plant Science，2022，13：853042.
[23]	王大伟.GmLUX调控大豆生育期的分子功能验证［D］.哈尔滨：中国科学院大学（中国科学院东北地理与农业生态研究所），2019.
	WANG D W.Molecular function verification of GmLUX regulating soybean growth period［D］.Harbin：Northeast Institute of Geography and Agroecology，Chinese Academy of Sciences，2019.
[24]	CHEN S F， ZHOU Y Q， CHEN Y R，et al.Fastp：an ultra-fast all-in-one FASTQ preprocessor［J］.Bioinformatics，2018，34（17）：i884-i890.
[25]	LOVE M I， HUBER W， ANDERS S.Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2［J］.Genome Biology，2014，15（12）：550.
[26]	SONG Q X， LI Q T， LIU Y F，et al.Soybean GmbZIP123 gene enhances lipid content in the seeds of transgenic Arabidopsis plants［J］.Journal of Experimental Botany，2013，64（14）：4329-4341.
[27]	CHENG D W， LIU Y Y， WANG Y，et al.Establishment of high-efficiency genome editing in white birch （Betula platyphylla Suk.）［J］.Plant Biotechnology Journal，2024，22（1）：7-9.
[28]	DING J H， WANG K J， PANDEY S，et al.Molecular advances in bud dormancy in trees［J］.Journal of Experimental Botany，2024，75（19）：6063-6075.
[29]	HAZEN S P， SCHULTZ T F， PRUNEDA-PAZ J L，et al.LUX ARRHYTHMO encodes a myb domain protein essential for circadian rhythms［J］.Proceedings of the National Academy of Sciences of the United States of America，2005，102（29）：10387-10392.
[30]	DAI S H， WEI X P， PEI L P，et al.BROTHER OF LUX ARRHYTHMO is a component of the Arabidopsis circadian clock［J］.The Plant Cell，2011，23（3）：961-972.
[31]	WANG J， LIAO X L， WU Z H，et al.Genetic control of seasonal meristem arrest in trees［J］.Proceedings of the National Academy of Sciences of the United States of America，2025，122（48）：e2505641122.
[32]	IBÁÑEZ C， KOZAREWA I， JOHANSSON M，et al.Circadian clock components regulate entry and affect exit of seasonal dormancy as well as winter hardiness in Populus trees［J］.Plant Physiology，2010，153（4）：1823-1833.
[33]	LIU D G， TANG D， XIE M，et al. Agave REVEILLE1 regulates the onset and release of seasonal dormancy in Populus ［J］.Plant Physiology，2023，191（3）：1492-1504.
[34]	VEERABAGU M， VAN DER SCHOOT C， TUREČK-OVÁ V，et al.Light on perenniality：para-dormancy is based on ABA-GA antagonism and endo-dormancy on the shutdown of GA biosynthesis［J］.Plant，Cell & Environment，2023，46（6）：1785-1804.
[35]	ZHENG C L， ACHEAMPONG A K， SHI Z W，et al.Abscisic acid catabolism enhances dormancy release of grapevine buds［J］.Plant，Cell & Environment，2018，41（10）：2490-2503.
[36]	ZHUANG W B， GAO Z H， WANG L J，et al.Comparative proteomic and transcriptomic approaches to address the active role of GA4 in Japanese apricot flower bud dormancy release［J］.Journal of Experimental Botany，2013，64（16）：4953-4966.
[37]	SINGH R K， MAURYA J P， AZEEZ A，et al.A genetic network mediating the control of bud break in hybrid aspen［J］.Nature Communications，2018，9：4173.
[38]	ANDRÉ D， ZAMBRANO J A， ZHANG B，et al. Populus SVL acts in leaves to modulate the timing of growth cessation and bud set［J］.Frontiers in Plant Science，2022，13：823019.
[39]	GÓMEZ-SOTO D， ALLONA I， PERALES M.FLOWERING LOCUS T2 promotes shoot apex development and restricts internode elongation via the 13-hydroxylation gibberellin biosynthesis pathway in poplar［J］.Frontiers in Plant Science，2022，12：814195.
[40]	TYLEWICZ S， PETTERLE A， MARTTILA S，et al.Photoperiodic control of seasonal growth is mediated by ABA acting on cell-cell communication［J］.Science，2018，360（6385）：212-215.
[41]	YUAN Y P， ZENG L L， KONG D R，et al.Abscisic acid-induced transcription factor PsMYB306 negatively regulates tree peony bud dormancy release［J］.Plant Physiology，2024，194（4）：2449-2471.
[42]	NAIR A， MAURYA J P， PANDEY S K，et al.ELF3 coordinates temperature and photoperiodic control of seasonal growth in hybrid aspen［J］.Current Biology，2025，35（7）：1484-1494.e2.
[43]	ZHONG M， ZENG B J， TANG D Y，et al.The blue light receptor CRY1 interacts with GID1 and DELLA proteins to repress GA signaling during photomorphogenesis in Arabidopsis ［J］.Molecular Plant，2021，14（8）：1328-1342.
[44]	CHEN K， LI G J， BRESSAN R A，et al.Abscisic acid dynamics，signaling，and functions in plants［J］.Journal of Integrative Plant Biology，2020，62（1）：25-54.
[45]	WU X M， WEI X R， LI Z，et al.Molecular cloning of cryptochrome 1 from Lilium × formolongi and the characterization of its photoperiodic flowering function in Arabidopsis ［J］.Plant Science，2022，316：111164.
[46]	LEE B D， KIM M R， KANG M Y，et al.The F-box protein FKF1 inhibits dimerization of COP1 in the control of photoperiodic flowering［J］.Nature Communications，2017，8：2259.
[47]	BABA K， KARLBERG A， SCHMIDT J，et al.Activity-dormancy transition in the cambial meristem involves stage-specific modulation of auxin response in hybrid aspen［J］.Proceedings of the National Academy of Sciences of the United States of America，2011，108（8）：3418-3423.
[48]	ZAWASKI C， KADMIEL M， PICKENS J，et al.Repression of gibberellin biosynthesis or signaling produces striking alterations in poplar growth，morphology，and flowering［J］.Planta，2011，234（6）：1285-1298.
[49]	HELFER A， NUSINOW D A， CHOW B Y，et al. LUX ARRHYTHMO encodes a nighttime repressor of circadian gene expression in the Arabidopsis core clock［J］.Current Biology，2011，21（2）：126-133.

引物名称 Primer name	引物序列（5′→3′） Primer sequences（5′→3′）
BpTUB-F	TCAACCGCCTTGTCTCTCAGG
BpTUB-R	TGGCTCGAATGCACTGTTGG
BpLUX1-RT-F	GTTCTCTGCCAGCGATAGCTTG
BpLUX1-RT-R	CGCGCAGTCGATTCTTCTAGACTT
BpLUX5-RT-F	TTGTCTTGGAGAACCTGCTTCGT
BpLUX5-RT-R	TGACAGGTAGGTCCATCTCAAGG
BpLUX18-RT-F	CATCATGCGGACAATGAATGTGAAG
BpLUX18-RT-R	TTGCACTTCCATCTGTGCTCTC

引物名称 Primer name	引物序列（5′→3′） Primer sequences（5′→3′）
BpLUX1-T1-F	TTCGTCGGCGTTCAGCATCTCT
BpLUX1-T1-R	AACCAGAGATGCTGAACGCCGA
BpLUX1-T2-F	TTCGTGGACGTTGTGGGCCACCT
BpLUX1-T2-R	AACAGGTGGCCCACAACGTCCAC
BpLUX5-T1-F	TTCGAGCTTGTGGGCCTTGAGA
BpLUX5-T1-R	AACTCTCAAGGCCCACAAGCTC
BpLUX5-T2-F	TTCGCAAGTTTGTTGCTGCTGTTC
BpLUX5-T2-R	AACGAACAGCAGCAACAAACTTGC
PSC1-F	TTGGCCGATTCATTAATGCAGC
Test-BpLUX1-T1-F	ATGATGGGTGAGGACGTGAAGATG
Test-BpLUX1-T1-R	CAAGCTATCGCTGGCAGAGAAC
Test-BpLUX1-T2-F	AAGTCTAGAAGAATCGACTGCGCG
Test-BpLUX1-T2-R	GACGACGCAAACAACTGGTGGTC