Research Article

Cloning and expression of an APETALA1-like gene from Nelumbo nucifera

Published: June 18, 2015
Genet. Mol. Res. 14 (2) : 6819-6829 DOI: 10.4238/2015.June.18.24

Abstract

The objective of this study was to clone the full-length cDNA of the APETALA1 (AP1) gene from lotus and analyze its sequence and expression pattern. The full-length cDNA sequence of the NnAP1 gene was amplified from the petals of Nelumbo nucifera ‘Hongxia’ using RT-PCR and rapid amplification of cDNA ends. Bioinformatic methods were used to analyze the sequence characteristics of the gene. Quantitative real-time PCR methods were used to investigate the expression pattern of NnAP1 in various organs and during different developmental stages. The cloned full-length NnAP1 cDNA (GenBank accession No. KF361315) was 902 bp, containing a 795-bp open reading frame encoding 264 amino acids with a relative molecular mass of 30,288.4 and an isoelectric point of 9.13. NnAP1 had a MADS-box domain and a K-box domain, which is typical of the SQUA/AP1 gene family. A protein sequence identity search showed that NnAP1 was 75-96% similar to other plant AP1s. Phylogenetic tree analysis indicated that NnAP1 was very closely related to AP1 of Glycine max, suggesting that they shared the same protein ancestor. Quantitative real-time PCR analysis showed that NnAP1 was expressed in various organs during different developmental stages; it had the highest expression in blooming flowers and had trace expression in the young vegetative and flower senescence stages. Our analysis suggests that NnAP1 plays an important role in controlling floral meristem identity and floral organ formation.

The objective of this study was to clone the full-length cDNA of the APETALA1 (AP1) gene from lotus and analyze its sequence and expression pattern. The full-length cDNA sequence of the NnAP1 gene was amplified from the petals of Nelumbo nucifera ‘Hongxia’ using RT-PCR and rapid amplification of cDNA ends. Bioinformatic methods were used to analyze the sequence characteristics of the gene. Quantitative real-time PCR methods were used to investigate the expression pattern of NnAP1 in various organs and during different developmental stages. The cloned full-length NnAP1 cDNA (GenBank accession No. KF361315) was 902 bp, containing a 795-bp open reading frame encoding 264 amino acids with a relative molecular mass of 30,288.4 and an isoelectric point of 9.13. NnAP1 had a MADS-box domain and a K-box domain, which is typical of the SQUA/AP1 gene family. A protein sequence identity search showed that NnAP1 was 75-96% similar to other plant AP1s. Phylogenetic tree analysis indicated that NnAP1 was very closely related to AP1 of Glycine max, suggesting that they shared the same protein ancestor. Quantitative real-time PCR analysis showed that NnAP1 was expressed in various organs during different developmental stages; it had the highest expression in blooming flowers and had trace expression in the young vegetative and flower senescence stages. Our analysis suggests that NnAP1 plays an important role in controlling floral meristem identity and floral organ formation.