Cloning of PhalPI Gene from Phalaenopsis and Its Expression in Floral Organ Mutants

doi:10.7525/j.issn.1673-5102.2017.03.012

Abstract

Abstract: For the regulatory mechanism of floral organ development in orchid, PhalPI gene(GenBank accession number:KY020416) was cloned from Phalaenopsis petals. By the sequence analysis, the cDNA full-length of this gene was 944 bp encoding 210 amino acid residues. The gene belongs to the GLO/PI group of the B class MADS-box transcription factor, which was highly identified with PhPI10 and PeMADS6 from Phalaenopsis. The PhalPI was expressed in reproductive organs and not in vegetative organs, the expression level was reduced in ovary after pollination. In 5 floral organ mutants, the expression level of the PhalPI gene was higher significantly in sepal and column of lip-like sepal mutants than that of the normal. The expression of this gene was decreased in sepals and petals in petal-like stamens mutant, and it was increased significantly in lip and column in this mutant. In column of Lateral petals pillared mutants, the expression of PhalPI gene was also increased significantly. The changes of expression level of the PhalPI gene in 3 floral organ mutants were associated with morphological mutations of floral organ. However, the expression levels of PhalPI gene were not changed in lip-like Lateral petals and anther-like Lateral petals mutants. Therefore, the PhalPI gene plays a crucial role in regulating Lateral petal and lip development in Phalaenopsis.

Key words: Phalaenopsis, GLO/PI, MADS-box, floral organ mutants

CLC Number:

S682.31

YUAN Xiu-Yun, XU Shen-Ping, WANG Ying-Bo, CUI Bo. Cloning of PhalPI Gene from Phalaenopsis and Its Expression in Floral Organ Mutants[J]. Bulletin of Botanical Research, 2017, 37(3): 416-423.

References

1. Litt A,Kramer E M. The ABC model and the diversification of floral organ identity[J]. Seminars in Cell & Developmental Biology,2010,21(1):129-137.
2. Kater M M,Dreni L,Colombo L. Functional conservation of MADS-box factors controlling floral organ identity in rice and Arabidopsis[J]. Journal of Experimental Botany,2006,57(13):3433-3444.
3. Airoldi C A,Davies B. Gene duplication and the evolution of plant MADS-box transcription factors[J]. Journal of Genetics and Genomics,2012,39(4):157-165.
4. Soltis P S,Soltis D E. Flower diversity and angiosperm diversification[M]. //Riechmann J L,Wellmer F. Flower development. New York:Springer,2014,1110:85-102.
5. Kanno A,Nakada M,Akita Y,et al. Class B gene expression and the modified ABC model in nongrass monocots[J]. The Scientific World Journal,2007,7:268-279.
6. Tsai W C,Chen H H. The orchid MADS-box genes controlling floral morphogenesis[J]. The Scientific World Journal,2006,6:1933-1944.
7. Su C L,Chen W C,Lee A Y,et al. A modified ABCDE model of flowering in orchids based on gene expression profiling studies of the moth orchid Phalaenopsis aphrodite[J]. PLoS One,2013,8(11):e80462.
8. Zahn L M,Leebens-mack J,Depamphilis C W,et al. To B or Not to B a flower:the role of DEFICIENS and GLOBOSA orthologs in the evolution of the angiosperms[J]. Journal of Heredity,2005,96(3):225-240.
9. Mondragón-palomino M,Theiβen G. Why are orchid flowers so diverse? Reduction of evolutionary constraints by paralogues of class B floral homeotic genes[J]. Annals of Botany,2009,104(3):583-594.
10. Guo B,Zhang T,Shi J L,et al. Cloning and characterization of a novel PI-like MADS-box gene in Phalaenopsis orchid[J]. DNA Sequence,2008,19(3):332-339.
11. Pan Z J,Cheng C C,Tsai W C,et al. The duplicated B-class MADS-box genes display dualistic characters in orchid floral organ identity and growth[J]. Plant and Cell Physiology,2011,52(9):1515-1531.
12. Mondragón-palomino M,Theiβen G. MADS about the evolution of orchid flowers[J]. Trends in Plant Science,2008,13(2):51-59.
13. Mondragón-palomino M,Theiβen G. Conserved differential expression of paralogous DEFICIENS- and GLOBOSA-like MADS-box genes in the flowers of Orchidaceae:refining the ‘orchid code’[J]. The Plant Journal,2011,66(6):1008-1019.
14. Becker A,Winter K U,Meyer B,et al. MADS-Box gene diversity in seed plants 300 million years ago[J]. Molecular Biology and Evolution,2000,17(10):1425-1434.
15. Vandenbussche M,Zethof J,Royaert S,et al. The duplicated B-class heterodimer model:whorl-specific effects and complex genetic interactions in Petunia hybrida flower development[J]. The Plant Cell,2004,16(3):741-754.
16. Soltis D E,Ma H,Frohlich M W,et al. The floral genome:an evolutionary history of gene duplication and shifting patterns of gene expression[J]. Trends in Plant Science,2007,12(8):358-367.
17. Irish V F. Evolution of petal identity[J]. Journal of Experimental Botany,2009,60(9):2517-2527.
18. Hernández-hernández T,Martínez-castilla L P,Alvarez-buylla E R. Functional diversification of B MADS-box homeotic regulators of flower development:adaptive evolution in protein-protein interaction domains after major gene duplication events[J]. Molecular Biology and Evolution,2007,24(2):465-481.
19. Panchy N,Lehti-shiu M,Shiu S H. Evolution of gene duplication in plants[J]. Plant Physiology,2016,171(4):2294-2316.
20. Ackerman C M,Yu Q Y,Kim S,et al. B-class MADS-box genes in trioecious papaya:two paleoAP3 paralogs,CpTM6-1 and CpTM6-2,and a PI ortholog CpPI[J]. Planta,2008,227(4):741-753.
21. Mondragón-palomino M,Hiese L,Härter A,et al. Positive selection and ancient duplications in the evolution of class B floral homeotic genes of orchids and grasses[J]. BMC Evolutionary Biology,2009,9:81.
22. Cantone C,Gaudio L,Aceto S. The PI/GLO-like locus in orchids:duplication and purifying selection at synonymous sites within Orchidinae(Orchidaceae)[J]. Gene,2011,481(1):48-55.
23. Melzer R,Härter A,Rümpler F,et al. DEF- and GLO-like proteins may have lost most of their interaction partners during angiosperm evolution[J]. Annals of Botany,2014,114(7):1431-1443.
24. Chang Y Y,Kao N H,Li J Y,et al. Characterization of the possible roles for B class MADS box genes in regulation of perianth formation in orchid[J]. Plant Physiology,2010,152(2):837-853.
25. Tsai W C,Kuoh C S,Chuang M H,et al. Four DEF-like MADS box genes displayed distinct floral morphogenetic roles in Phalaenopsis orchid[J]. Plant and Cell Physiology,2004,45(7):831-844.
26. Tsai W C,Pan Z J,Hsiao Y Y,et al. Interactions of B-class complex proteins involved in tepal development in Phalaenopsis orchid[J]. Plant and Cell Physiology,2008,49(5):814-824.
27. Pan Z J,Chen Y Y,Du J S,et al. Flower development of Phalaenopsis orchid involves functionally divergent SEPALLATA-like genes[J]. New Phytologist,2014,202(3):1024-1042.
28. Tsai W C,Lee P F,Chen H I,et al. PeMADS6,a GLOBOSA/PISTILLATA-like gene in Phalaenopsis equestris involved in petaloid formation,and correlated with flower longevity and ovary development[J]. Plant and Cell Physiology,2005,46(7):1125-1139.
29. Guo B,Hexige S,Zhang T,et al. Cloning and characterization of a PI-like MADS-box gene in Phalaenopsis orchid[J]. Journal of Biochemistry and Molecular Biology,2007,40(6):845-852.
30. Salemme M,Sica M,Gaudio L,et al. Expression pattern of two paralogs of the PI/GLO-like locus during Orchis italica(Orchidaceae,Orchidinae) flower development[J]. Development Genes and Evolution,2011,221(4):241-246.
31. Mao W T,Hsu H F,Hsu W H,et al. The C-terminal sequence and PI motif of the Orchid(Oncidium Gower Ramsey) PISTILLATA(PI) ortholog determine its ability to bind AP3 orthologs and enter the nucleus to regulate downstream genes controlling petal and stamen formation[J]. Plant and Cell Physiology,2015,56(11):2079-2099.
32. Xu Y F,Teo L L,Zhou J,et al. Floral organ identity genes in the orchid Dendrobium crumenatum[J]. The Plant Journal,2006,46(1):54-68.
33. Chen Y Y,Lee P F,Hsiao Y Y,et al. C- and D-class MADS-box genes from Phalaenopsis equestris(Orchidaceae) display functions in gynostemium and ovule development[J]. Plant and Cell Physiology,2012,53(6):1053-1067.
34. Huang J Z,Lin C P,Cheng T C,et al. A de novo floral transcriptome reveals clues into Phalaenopsis orchid flower development[J]. PLoS One,2015,10(5):e0123474.

[1]	Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG. Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia [J]. Bulletin of Botanical Research, 2021, 41(6): 888-895.
[2]	TAN Chuan-Ting, SONG Wei-Han, ZHU Ming-Ku, XU Tao, DONG Ting-Ting, LI Zong-Yun. Cloning and Expression Analysis of Stress Treatments of a MADS-box Transcription Factor in Sweet Potato [J]. Bulletin of Botanical Research, 2017, 37(4): 587-595.
[3]	ZU Kui-Ling, DONG Shu-Bin, LI Jian-Xia, ZHAO Yun-Yu, ZHAO Liang-Cheng. Sequence and Expression Analysis of MADS-box Genes in Celastrus orbiculatus Thunb. Based on RNA-seq Data [J]. Bulletin of Botanical Research, 2017, 37(1): 69-77.
[4]	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.