Isolation and Functional Analysis of FLC Gene from Poplar (Populus simonii×Populus nigra)

doi:10.7525/j.issn.1673-5102.2015.03.007

Abstract

Abstract: MADS-box family protein is an important class of transcription factors regulating flowering. FLC is a member of MADS-box transcription factors, and the FLC gene is controlled by many pathways, and then to regulate the flowering. We cloned a FLC-homolog gene in male flower buds from Populus simonii×Populus nigra. The open reading frame (ORF) of this gene had 726 bp which encoded a protein containing 241 amino acid. The molecular weight of the predicted protein was 27.559 KD, and the pI was 9.37. By real-time quantitative PCR, vernalization could down-regulate the expression of PnFLC 57.9%-84% in the root, stem and leaves tissues of poplar. By GUS staining, the promoter of the PnFLC is highly expressed in actively dividing tissues. Furthermore, by transforming Arabopsis, PnFLC down-regulated the expression of AtAP1, AtSOC1 and AtFT, while delayed the flowering in Arabidopsis.

Key words: Populus simonii×, Populus nigra, FLC gene, flowering, vernalization

CLC Number:

SUN Qi；QU Chun-Pu；ZHENG Mei-Zhu；LIU Guan-Jun. Isolation and Functional Analysis of FLC Gene from Poplar (Populus simonii×Populus nigra)[J]. Bulletin of Botanical Research, 2015, 35(3): 363-369.

References

1.Feng W,Michaels S D.Dual roles for FY in the regulation of FLC［J］.Plant Signaling & Behavior,2011,6(5):703-705.

2.Michaels S D,Amasino R M.Loss of FLOWERING LOCUS C activity eliminates the late-flowering phenotype of FRIGIDA and autonmous pathway mutations but not responsiveness to vernalization［J］.The Plant Cell,2001,13(4):935-941.

3.He Y H,Amasino R M.Role of chromatin modification in flowering-time control［J］.Trends in Plant Science,2005,10(1):30-35.

4.Macknight R,Bancroft I,Page T,et al.FCA,a gene controlling flowering time in Arabidopsis,encodes a protein containing RNA-Binding domains［J］.Cell,1997,89(5):737-745.

5.Lee I,Aukerman M J,Gore S L,et al.Isolation of LUMINIDEPENDENS:A gene involved in the control of flowering time in Arabidopsis［J］.The Plant Cell,1994,6(1):75-83.

6.Michaels S D,Amasino R M.FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering［J］.The Plant Cell,1999,11(5):949-956.

7.Rouse D T,Sheldon C C,Bagnall D J,et al.FLC,a repressor of flowering,is regulated by genes in different inductive pathways［J］.Plant Journal,2002,29(2):183-191.

8.Helliwell C A,Wood C C,Robertson M,et al.The Arabidopsis FLC protein interacts directly in vivo with SOC1 and FT chromatin and is part of a high-molecular weight protein complex［J］.Plant Journal,2006,46(2):183-192.

9.Lin S I,Wang J G,Poon S Y,et al.Differential regulation of FLOWERING LOCUS C expression by vernalization in Cabbage and Arabidopsis［J］.Plant Physiology,2005,137(3):1037-1048.

10.Sheldon C C,Rouse D T,Finnegan E J,et al.The molecular basis of vernalization:The central role of FLOWERING LOCUS C(FLC)［J］.Proc Nati Acad Sci USA,2000,97(7):3753-3758.

11.Fang Q,Xu Z,Song R.Cloning,characterization and genetic engineering of FLC homolog in Thellungiella halophila［J］.Biochemical and Biophysical Research Communications,2006,347(3):707-714.

12.张勇,张守功,齐力旺,等.杨树—林木基因组学研究的模式物种［J］.植物学报,2006,23(3):286-293.

13.Tuskan GA,Difazio S,Jansson S,et al.The genome of black cottonwood,Populus trichocarpa(Torr.& Gray)［J］.Science,2006,313(5793):1596-1604.

14.Clough S J,Bent A F.Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana［J］.Plant Journal,1998,16(6):735-743.

15.Brand L,Hörler M,Nüesch E,et al.A versatile and reliable two-component system for tissue-specific gene［J］.Plant Physiology,2006,141(4):1194-1204.

16.Kim S Y,Park B S,Kwon S J,et al.Delayed flowering time in Arabidopsis and Brassica rapa by the overexpression of FLOWERING LOCUS C(FLC) homologs isolated from Chinese cabbage(Brassica rapa L.ssp.pekinensis)［J］.Plant Cell Rep,2007,26(3):327-336.

17.Sheldon C C,Burn J E,Perez P P.The FLF MADS box gene:A repressor of flowering in Arabidopsis regulated by vernalization and methylation［J］.The Plant Cell,1999,11(3):445-458.

18.Sheldon C C,Rouse D T,Finnegan E J,et al.The molecular basis of vernalization:The central role of FLOWERING LOCUS C(FLC)［J］.Proc Nati Acad Sci USA,2000,97(7):3753-3758.

19.朱芹芹,李忠爱,何艳霞,等.表观遗传与花期调控研究进展［J］.园艺学报,2013,40(9):1787-1794.

20.Chiang G C,Barua D,Kramer EM,et al.Major flowering time gene,FLOWERING LOCUS C,regulates seed germination in Arabidopsis thaliana［J］.Proc Nati Acad Sci USA,2009,106(28):11661-11666.

[1]	Changrui TAI, Kai ZHAO, Yanfang YANG, Yan WU, Wei ZHAI, Yuewei TONG. Flowering Biological Characteristics and Breeding System of the Rare and Endangered Plant of Sinojackia microcarpa [J]. Bulletin of Botanical Research, 2023, 43(2): 311-320.
[2]	Jianwei YANG, Zongyan LI, Yao FENG, Shuxian REN, Menglu HU, Songpu YE. Reproductive Biological Characteristics of Dendrobium chrysanthum [J]. Bulletin of Botanical Research, 2023, 43(1): 150-160.
[3]	Liben PAN, Xue YAN, Jia LIU, Kexin WU, Yang LIU, Shaochong LIU. Physiological Characteristics of Early Spring Flowering Plants under Northeast Forest [J]. Bulletin of Botanical Research, 2022, 42(4): 657-666.
[4]	CHEN Xiang-Bo, LÜ Xiu-Li, LIU Yang, ZHAO Ming-Shui, CUI Xin-Hong, ZHANG Dong-Mei. Floral Morphology and Flowering Process of Acer yangjuechi, the Extremely Endangered Plant [J]. Bulletin of Botanical Research, 2019, 39(3): 329-337.
[5]	JIANG Zheng, SUN Li-Yong, LIU Xu, LIU Chen-Ni, YIN Zeng-Fang. Nutritional Effect and Rhythm of Spring and Summer Flowering in Magnolia soulangeana‘Changchun’ [J]. Bulletin of Botanical Research, 2019, 39(2): 192-199.
[6]	WANG Xi-Gang, ZHAN Ya-Guang, ZHANG Gui-Qin, YAN Chao-Fu, LI Xiang, ZENG Fan-Suo. Influences of Fertilization on Growth and Fruiting of Fraxinus mandschurica [J]. Bulletin of Botanical Research, 2017, 37(2): 298-303.
[7]	KANG Xiao-Shan, Sophia, DUAN Shi-Min, PAN Bo-Rong, ZHANG Yong-Zhi, TIAN Cong. Preliminary Studies on Hybridization among Sympatric Distributed Calligonum Species in Northern Xinjiang [J]. Bulletin of Botanical Research, 2016, 36(5): 790-794.
[8]	LI Ya-Lan, LIANG Feng-Li. Stamen Dynamics and Breeding System of Lonicera alberti [J]. Bulletin of Botanical Research, 2016, 36(4): 503-510.
[9]	WANG Xiao-Dong;HE Guo-Qiang;ZHAO Li-Qun;HU Yan-Bo;LI Chang-Hai*. Effects of Shading on the Photosynthesis Characteristics and Fruit Traits of Laiweili Raspberry [J]. Bulletin of Botanical Research, 2014, 34(5): 599-603.
[10]	ZHENG Xing-Xing;WANG Tie-Juan;WEI Xiao-Meng. Comparison of Flowering Phenology of Two Kinds of Sandy Semi-shrub of Artemisia* L. in the Horqin Sandy Land [J]. Bulletin of Botanical Research, 2014, 34(4): 446-451.
[11]	YUAN Xiu-Yun;TIAN Yun-Fang;JIANG Su-Hua;WANG Mo-Fei;MA Jie;CUI Bo. Cloning and Expression Analysis of DtpsMADS1* Gene from Doritaenopsis hybrid [J]. Bulletin of Botanical Research, 2014, 34(1): 53-61.
[12]	SUN Hui-Min;GU Xiao-Ping;YUAN Jin-Ling;YUE Jin-Jun. Analysis on DNA Methylation-sensitive Amplified Polymorphism in Phyllostachys bambusoides* During Flowering and Rejuvenation [J]. Bulletin of Botanical Research, 2013, 33(6): 723-730.
[13]	LI Jin-Xia;SU Xue;LIU Yu-Guo;CHEN Ming-Zhong;SUN Kun. Flowering Dynamics and Breeding System of Incarvillea sinensis* var. przewalskii [J]. Bulletin of Botanical Research, 2011, 31(2): 147-151.
[14]	SONG Yu-Xia;GUO Sheng-Hu;NIU Dong-Ling;ZHENG Guo-Qi;MA Hong-Ai;LI Miao. A Study on Breeding System of the Endangered Plant Cistanche deserticola [J]. Bulletin of Botanical Research, 2008, 28(3): 278-282.
[15]	SUN Yan, WANG Chen, LIU Ming-Yuan. THE BIOLOGICAL CHARACTERS OF FLOWERING AND FRUITING OF GUANLONGDAN [J]. Bulletin of Botanical Research, 2003, 23(4): 453-457.