Cloning,Expression and Vector Construction of a MADS-box Gene in Chrysanthemum morifolium

doi:10.7668/hbnxb.2016.03.004

Abstract

Abstract: In order to research the identification of gene function and genetic improvement in Chrysanthemum,a MADS-box family class A gene was isolated from the leaf of Chrysanthemum morifolium by using RT-PCR method,which was designated CmFUL gene(The accession number in GenBank:KT894379).The coding region of this gene(ORF) was 741 bp and encoded 246 putative amino acid residues.Encoding product of CmFUL gene was a kind of hydrophilic protein and predicted that the protein contained 12 phosphorylation sites and 2 potential N-Glycosylation sites.Sequence and phylogenic analyses revealed that CmFUL was close to CDM41 with 95.6% sequence similarity which both belonged to the FUL -like clade of AP1/FUL subfamily.The expression analyses of CmFUL by Real-time fluorescent quantitative PCR indicated that this gene was expressed in different parts in bloom stage,but the expression abundance was varied widely.The highest amount of expression was observed in buds,followed by in tubiform florets.The expression level in roots and lingulate florets were lowest.It was worth noting that the level expressed in tubiform florets was 12 times of that in lingulate florets between a same flower.The results implied that CmFUL might play an important role in promoting flowering and ovary development.CmFUL was linked to pCAMBIA1300 vector and the plant expression vector was constructed successfully.

Key words: Chrysanthemum morifolium, FUL -like, Gene cloning, Expression analysis, Bioinformatics

CLC Number:

Q949.71+8.43

LIANG Fang, XU Shenping, JIANG Suhua, YUAN Xiuyun, CUI Bo, ZHANG Jing. Cloning,Expression and Vector Construction of a MADS-box Gene in Chrysanthemum morifolium[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(3): 25-31. doi: 10.7668/hbnxb.2016.03.004.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2016.03.004

http://www.hbnxb.net/EN/Y2016/V31/I3/25

References

[1] Alvarez-Buylla E R,Pelaz S,Liljegren S J,et al.An ancestral MADS-box gene duplication occurred before the divergence of plants and animals[J].Proceedings of National Academy Sciences,2000,97(10):5328-5333.
[2] Chi Y,Huang F,Liu H C,et al.An APETALA1-like gene of soybean regulates flowering time and specifies floral organs [J].Journal of Plant Physiology,2011,168(18):2251-2259.
[3] Galimba K D,Di Stilio V S.Sub-functionalization to ovule development following duplication of a floral organ identity gene [J].Development Biology,2015,405(1):158-172.
[4] Yu C,Liu Y,Zhang A,et al.MADS-box transcription factor OsMADS25 regulates root development through affection of nitrate accumulation in rice[J].PLoS One,2015,10(8):e0135196.
[5] Ferrándiz C,Liljegren S J,Yanofsky M F.Negative regulation of the SHATTERPROOF genes by FRUITFFULL during Arabidopsis fruit development[J].Science,2000,289(5478):436-438.
[6] Shima Y,Kitagawa M,Fujisawa M,et al.Tomato FRUITFULL homologues act in fruit ripening via forming MADS-box transcription factor complexes with RIN[J].Plant Molecular Biology,2013,82(4-5):427-438.
[7] Voogd C,Wang T,Varkonyi-Gasic E.Functional and expression analyses of kiwifruit SOC1-like genes suggest that they may not have a role in the transition to flowering but may affect the duration of dormancy[J].Journal of Experimental Botany,2015,66(15):4699-4710.
[8] Xie Q,Hu Z,Zhu Z,et al.Overexpression of a novel MADS-box gene SlFYFL delays senescence,fruit ripening and abscission in tomato[J].Scientific Reports,2014,4:4367.
[9] Hoenicka H,Nowitzki O,Hanelt D,et al.Heterologous overexpression of the birch FRUITFULL -like MADS-box gene BpMADS4 prevents normal senescence and winter dormancy in Populua tremula L.[J].Planta,2008,227(5):1001-1011.
[10] Ferrándiz C,Gu Q,Martienssen R,et al.Redundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1 and CAULIFLOWER [J].Development,2000,127(4):725-734.
[11] Mandel M A,Gustafson-Brown C,Savidge B,et al.Molecular characterization of the Arabidopsis floral homeotic-gene, APETALA1 [J].Nature,1992,360(6401):273-277.
[12] Kempin S A,Savidge B,Yanofsky M F,et al.Molecular basis of the cauliflower phenotype in Arabidopsis [J].Science,1995,267(5197):522-25.
[13] Mandel M A,Yanofsky F M.The Arabidopsis AGL8 MADS box gene is expressed in inflorescence meristems and is negatively regulated by APETALA1 [J].The Plant Cell,1995,7(11):1763-1771.
[14] Torti S,Fornara F.AGL24 acts in concert with SOC1 and FUL during Arabidopsis floral transition [J].Plant Signaling and Behavior,2012,7(10):1251-1254.
[15] Melzer S,Lens F,Gennen J,et al.Flowering-time genes modulate meristem determinacy and growth form in Arabidopsis thaliana [J].Nature Genetics,2008,4(12):1489-1492.
[16] Pabón-Mora N,Sharma B,Holappa L D,et al.The Aquilegia FRUITFULL-like genes play key roles in leaf morphogenesis and inflorescence development[J].Plant Journal,2013,74(2):197-212.
[17] Gu Q,Ferr ndiz C,Yanofsky M F,et al.The FRUITFULL MADS-box gene cell differentiation during Arabidopsis fruit development[J].Development,1998,125(8):1509-1517.
[18] Xu Y,Zhang L,Ma R C.Functional characterization and mapping of two MADS box genes from peach ( Prunus persica )[J].Chinese Science Bulletin,2008,53(6):853-859.
[19] Zhang W X,Fan S L,Pang C Y,et al.Molecular cloning and function analysis of two SQUAMOSA-like MADS-box genes from Gossypium hirsutum L.[J].Journal of Intergrative Plant Biology,2013,55(7):597-607.
[20] Jia Z,Jiang B J,Gao X W,et al. GmFULa, a FRUITFULL homolog,functions in the flowering and maturation of soybean[J].Plant Cell Reports,2015,34(1):121-132.
[21] Shchennikova V A,Shugla A O,Immink R,et al.Identification and characterization of four Chrysanthemum MADS-Box genes,belonging to the APETALA1/FRUITFULL and SEPALLATA3 subfamilies[J].Plant Physiology,2004,134(4):1632-1641.
[22] Goloveshkina N E,Shchennikova V A,Kamionshaya M A,et al.Influence of ectopic expression of Asteraceae MADS box genes on plant ontogeny in tobacco[J].Plant Cell Tissue and Organ Culture,2012,109(1):61-71.
[23] Wang Y,Ma Y P,Fu J X,et al.Isolation and functional analysis of the ClM8-FRUITFULL-like MADS-box gene from Chrysanthemum lavandulifolium [J].Scientia Horticulturae,2013,161(2):125-133.
[24] Litt A,Irish F V.Duplication and diversification in the APETALA1/FRUITFULL floral homeotic gene lineage:implications for the evolution of floral development[J].Genetics,2003,165(2):821-833.
[25] Shan H Y,Zhang N,Liu C J,et al.Patterns of gene duplication and functional diversification during the evolution of the AP1/SQUA subfamily of plant MADS-box genes[J].Molecular Phylogenetics and Evolution,2007,44(1):26-41.

Please choose a citation manager

Content to export