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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2011, Vol. 5 Issue (3) : 366-371     DOI: 10.1007/s11703-011-1095-1
RESEARCH ARTICLE |
Molecular cloning and sequence analysis of an LFY homologous gene from Juglans regia L.
Fuqiang HE, Hongxia WANG, Zhihua ZHANG()
Mountainous Areas Research Institute, Agricultural University of Hebei, Baoding 071001, China
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Abstract  

The existence of a long juvenile phase is found seriously to affect the early-stage economic benefits of later mature walnuts (Juglans regia L.). Studies on LFY, a key gene controlling floral transition and flower differentiation, may be helpful in solving the problem. This study reports the identification and characterization of a JrLFY gene from Juglans regia L., a homolog of FLORICAULA/LFY. The gene was isolated from flower buds of precocious walnut cultivar Zhonglin No. 5 by RT-PCR and RACE. The cDNA sequence of JrLFY (GenBank accession no. GU194836) was 1496 bp and contained an ORF of 1158 bp. Its corresponding genomic sequence (GenBank accession no. HQ019159) showed that the JrLFY contained three exons and two introns. The predicted amino acid sequence of the gene consisted of 385 amino acids and had a conserved region in the C-terminal when being aligned with sequences of other LFY homologs. Phylogenetic analysis showed that the LFY protein of walnut was close to those of hickory and chestnut. These studies will lay a foundation for understanding the mechanism of early fruiting and preparation for transfer of the JrLFY as a transgene to later mature walnuts.

Keywords walnut      JrLFY      early mature      precocious      juvenility     
Corresponding Authors: ZHANG Zhihua,Email:shylg@hebau.edu.cn   
Issue Date: 05 September 2011
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1095-1     OR     http://academic.hep.com.cn/fag/EN/Y2011/V5/I3/366
Fig.1  Schematic representation of gene. a stands for schematic diagram of the JrLFY cDNA ORF (black box) with 3'UTR (thin lines); b stands for exon–intron organization of the gene with location of exons (black boxes) and introns (thin lines). The number 1 indicates the location of the translational start codon as determined. Numbers delineate addresses in nucleic acid sequences.
OrganismAccession numberNumber of intronsLength (bp)Number of exonsLength (bp)Number of amino acid residues
Juglans regiaHQ0191592545, 1,1663433, 362, 363385
Arabidopsis thalianaAF4667922458, 9073457, 407, 411424
Solanum tuberosumEU3710472600, 5723477, 407, 378418
Solanum lycopersicumAF1979362309, 4093475, 389, 375412
Capsicum annuumEU0002542712, 2,0203457, 413, 381416
ChrysanthemumlavandulifoliumAY6725422583, 1,0833463, 386, 390412
Boechera strictaAY7345642453, 1,1753430, 404, 355396
Orchis italicaAB08885121,070, 1203486, 444, 412453
Vitis viniferaAF3781262153, 6633451, 377, 381402
Citrus sinensisAY3389762408, 6753463, 353, 381398
Idahoa scapigeraAY2192292556, 1,0703445, 389, 333389
Titanotrichum oldhamiiAY5263192421, 873433, 377, 369392
Serapias linguaAB0884662100, 1003492, 438, 438455
Cedrela fissilisAY6336212176, 6553450, 378, 363396
Arabidopsis lyrataAF4668023441, 76, 8764457, 35, 390, 411430
Solanum tuberosumEF0623572600, 5723477, 407, 373418
Tab.1  Organization of introns and exons in genomic sequence from different plants
Fig.2  Alignment of amino acid sequence of LFY (JrLFY, ACZ48701) with those of LFY (CcLFY, ABI58284), LFY (AMAFLO, AAA62574), FL2 (NtFL2, Q40505) and LFY (AtLFY, NM_125579). Identical amino acid residues in this alignment are shaded in black, and similar amino acid residues are shaded in red (higher similarity) and blue (lower similarity). The alignment was performed with the DNAMAN program.
Fig.3  Phylogenetic analysis of the 29 LFY proteins. Individual data are derived from GenBank except for provided by this research. The tree was displayed as a phylogram in which branch lengths are proportional to distance.
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[1] Baoguo LI, Suping GUO, Guohui QI. The main factors causing “imperfect shell development” (ISD) in thin-shelled walnut[J]. Front Agric Chin, 2009, 3(1): 75-77.
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