Molecular cloning, characterization and expression analysis of MADS-box genes associated with reproductive development in Momordica dioica Roxb.
- 81 Downloads
The repertoire and functions of MADS-box family transcription factors (TFs) largely remains unexplored with respect to floral organogenesis of Momordica dioica Roxb. Degenerative PCR followed by rapid amplification of cDNA ends was employed in the present study to clone and characterize 17 MADS-box genes (designated as MdMADS01 to MdMADS17) from the floral buds of M. dioica. The cloned genes were clustered into three subgroups (11 MIKCC, 4 MIKC* and 2 Mα) based on phylogenetic relationships with the MADS-box genes from Cucumis sativus, Cucumis melo and Arabidopsis thaliana. Southern hybridization showed that all the isolated genes were represented by single copy locus in the M. dioica genome. Gene structure analysis revealed 1–8 exons in MdMADS-box genes with the number of exons in MIKC greatly exceeding from that in M-type genes. Motif elicitation of the MdMADS-box genes indicated the presence of additional domains with MIKC type, suggesting that they had more complex structures. Expression analysis of MdMADS genes in six M. dioica transcriptome suggested that, 11 MIKCC—type genes are associated with floral homeotic functions, 4 MIKC*-type genes (MdMADS12 to MdMADS15) controlled the growth of male gametophyte, while the two M-type genes (MdMADS16 and MdMADS17) played significant role in female gametogenesis and seed development. Overall, these are the first set of MADS-box genes from M. dioica exhibiting a differential expression pattern during floral development. The results from this study will provide valuable information for further functional studies of candidate MADS-box genes in the sexual dimorphism of this economically important dioecious cucurbit.
KeywordsMomordica dioica L. MADS-box MIKC Floral development Expression analysis
The study is funded by a research grant (BT/PR3919/PBD/16/959/2011) from the Department of Biotechnology, Government of India. JNM is grateful to DBT, India for financial support in the form of Senior Research Fellowship. The authors are thankful to Prof. Manoj Ranjan Nayak, President, Siksha O Anusandhan University for his guidance and support. We also thank Prof. Sumita Jha, Centre for Advanced Study, Dept. of Botany, Calcutta University and Prof. Sanghamitra Nayak, Head, Centre of Biotechnology, Siksha O Anusandhan University for their able guidance and support. We also thank DST-FIST, Govt. of India, for the research infrastructure facilities provided to Centre of Biotechnology, Siksha O Anusandhan University.
RKJ conceived and supervised the project. JNM collected samples, isolated RNA, performed cloning, sequencing and qPCR expression analyses. JNM and RKJ interpreted the data and prepared the manuscript. Both authors read and approved the final manuscript.
Compliance with ethical standards
The authors have declared that no competing interest exists.
- Baratakke RC, Patil CG (2009) Karyomorphological investigations in dioecious climber Momordica dioica Roxb. J Cytol Genet 11:91–96Google Scholar
- Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:1241–1251Google Scholar
- Hossain MA, Islam M, Ali M (1996) Sexual crossing between two genetically female plants and sex genetics of kakrol (Momordica dioica Roxb.). Euphytica 90:121–125Google Scholar
- Litt A, Irish V (2003) Duplication and diversification in the APETALA/FRUITFUL floral homeotic gene lineage: implications for the evolution of floral development. Genetics 165:821–833Google Scholar
- Parenicová L, de Folter S, Kieffer M, Horner DS, Favalli C, Busscher J, Cook HE, Ingram RM, Kater MM, Davies B, Angenent GC, Colombo L (2003) Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis: new openings to the MADS world. Plant Cell 15:1538–1551CrossRefGoogle Scholar
- Tapia-López R, García-Ponce B, Dubrovsky JG, Garay-Arroyo A, Pérez- Ruíz RV, Kim SH, Acevedo F, Pelaz S, Alvarez-Buylla ER (2008) An AGAMOUS-related MADS-box gene, XAL1 (AGL12), regulates root meristem cell proliferation and flowering transition in Arabidopsis. Plant Physiol 146:1182–1192CrossRefGoogle Scholar