Cloning, Bioinformatics and Expression Analysis of Mango MiCOL6 Gene

doi:10.3969/j.issn.1000-2561.2020.04.012

Abstract

Abstract:

CONSTANS (CO) is a core regulator of the photoperiod pathway, which integrates light quality and clock output signals to regulate the floral transformation in plants. A full-length cDNA of MiCOL6 gene was obtained from transcriptome data analysis and then verified by RT-PCR amplification in mango. Sequences analysis showed that the open reading frame was 678 bp in length, coding 226 amino, the molecular weight and isoelectric point (pI) of MiCOL6 was 26.88 kDa and 4.85, respectively; and MiCOL6 was a non-secreted hydrophilic protein. Conserved domain and phylogenetic tree analysis showed that MiCOL6 contained only one CCT domain and belonged to the group IV of the CO gene family. The secondary structure analysis showed that the protein mainly consisted of random curl and α-helix. The subcellular localization prediction showed that MiCOL6 was likely to be localized in the cytoplasm membrane. Expression pattern analysis showed that the MiCOL6 was the highest expressed in leaves during the bud differentiation period. The results of the experiment would provide a theoretical basis for the further study on the function of MiCOL6 in mango.

Key words: mango, CONSTANS, MiCOL6, gene cloning, bioinformatics analysis, expression pattern analysis

CLC Number:

S667.7

LU Xinxi,LUO Cong,ZHANG Xiujuan,YU Haixia,LIU Yuan,HE Xinhua. Cloning, Bioinformatics and Expression Analysis of Mango MiCOL6 Gene[J]. Chinese Journal of Tropical Crops, 2020, 41(4): 715-721.

Figures/Tables 10

Tab. 1 Primers sequence

引物名称 Primers ID	引物序列（5°-3°） Primers sequence (5°-3°)
MiCOL6u	AAATGGGGTACCATTCATTGT
MiCOL6d	CTAAGCTTCTCAGGAAACTCT
qCOL6u	CTTATGGTGAAGAGGCCATGA
qCOL6d	CCAGTCCATGAAATCCCAAC
qActin1u	CCGAGACATGAAGGAGAAGC
qActin1d	GTGGTCTCATGGATACGAGCA

Fig. 1 Gel electrophoresis of PCR amplified products of MiCOL6 M: DL2000 marker; 1: PCR amplification product.

Fig. 2 Phylogenetic tree of CO proteins The GenBank accession numbers of the amino acids encoding each gene are as follows: Arabidopsis thaliana AtCOL (NM_121589), AtCOL1 (NM_ 121589), AtCOL2 (NM_111105), AtCOL6 (NM_ 106047), AtCOL7 (NM_103803), AtCOL9 (AB023039), AtCOL10 (NM_117613), AtCOL11 (NM_ 113084), AtCOL12 (NM_130356), AtCOL16 (NM_ 102355); Populus trichocarpa PtCOL16 (XP_024449388), Theobroma cacao TaCOL8 (XP_007016977).

Fig. 3 Multiple alignments of MiCOL6 with other species The GenBank accession numbers of the amino acids encoding each gene are as follows: Populus trichocarpa PtCOL16 (XP_024449388), Quercus suber QsCOL7 (XP_023874898), Gossypium arboretum GaCOL16 (XP_017641001).

Fig. 4 Conserved domain prediction of MiCOL6

Fig. 5 Signal peptide prediction of MiCOL6

Fig. 6 Transmembrane domain prediction of MiCOL6

Fig. 7 Phosphorylation site analysis of MiCOL6

Fig. 8 Hydrophobicity prediction of MiCOL6

Fig. 9 MiCOL6 expression at different flower development stages

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