“Isolation and Characterization of MIKC group MADS Box genes in Petuniaaxillaris”
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Date
2019
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SARDARVALLABH BHAI PATEL UNIVERSITY OF AGRICULTURE MEERUT 250110
Abstract
MADS-box gene family plays an important role in the development of floral organs in plants. A coding region of MADS-box genes, named as FBP20, FBP21, FBP22 and FBP28 were cloned by PCR from flower bud of Petunia axillaris. The coding sequence of FBP20, FBP21, FBP22 and FBP28 contained an ORF of 651, 657, 657 and 645 bp encoding 216, 218, 218 and 214 amino acid residues respectively. Protein multi-alignment analysis indicated that FBP20, FBP21, FBP22 and FBP28 shared high homology with other SOC1 family proteins from other plants. FBP20 and FBP21 showed high sequence identity to Nicotiana tabacum TobMADS1 with 87.93% (51/58) and 98.27% (57/58) respectively, FBP22 showed high sequence identity to Solanum lycopersicum SOC1 with 86.20% (50/58) and Arabidopsis thaliana AGL42 with 84.48% (49/58) and FBP28 showed high sequence identity to Antirrhinum majus DEFH68 with 74.13% (43/58) of amino acids identical in MADS-box domain. Moreover, FBP20, FBP21, FBP22 and FBP28 contained a MADS-box and a K-box domain, typical conservative domains of plant MADS-box proteins. In addition, a highly conserved SOC1 motif (DVETELFIGPP) for most SOC1-like genes was also identified at the C-terminal region of FBP20, FBP21, FBP22 and FBP28. Interestingly, FBP22 showed little variation in SOC1 motif. Phylogenetic tree clearly showed that FBP20, FBP21, FBP22 and FBP28 are classed into SOC1/TM3 subgroup of MADS-box protein family. On the basis of PCR analysis we revealed that the expression level of FBP20, FBP21, FBP22 and FBP28 genes in young flower bud were significantly higher than those in different other stages of flower and that FBP20, FBP21, FBP22 and FBP28 expression level increased only at young bud stage of flower development, suggesting that FBP20, FBP21, FBP22 and FBP28 are involved in floral transition and early flower development. The present study is also aim for molecular modelling of FBP20, FBP21, FBP22 and FBP28 by using bioinformatics applications. Development of Full FBP20, FBP21, FBP22 and FBP28 gene construct (pCAMBIA1301:FBP20, pCAMBIA1301:FBP21, pCAMBIA1301: FBP22 and
pCAMBIA1301:FBP28 respectively) as a transgene source based sequence homology analysis. Validation of these transgene construct pCAMBIA1301:FBP20, pCAMBIA1301:FBP21, pCAMBIA1301:FBP22 and pCAMBIA1301:FBP28 based forits
structural integrity through PCR amplification of different regulatory sequences from the construct as well as restriction digestion analysis. The Agrobacterium mediated host cells transformation activity of the 35S:FBP21 and 35S:FBP22 transgene in in-vitro plants, is confirmed with PCR genotyping.
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“Isolation and Characterization of MIKC group MADS Box genes in Petuniaaxillaris”
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“Isolation and Characterization of MIKC group MADS Box genes in Petuniaaxillaris”