Cloning,Subcellular Localization and Expression Analysis of MrAGL8 Gene in Medicago ruthenica

doi:10.7668/hbnxb.20195685

Abstract

Abstract:

In order to explore the relationship between MrAGL8 gene and pod dehiscence traits in Medicago ruthenica. In this study,Medicago ruthenica was used as the plant material.The MrAGL8 gene was amplified by PCR,cloned,and sequenced.Additionally,bioinformatics analysis,subcellular localization,and expression analysis in different tissues and organs were performed for this gene.The results showed that the complete coding region of MrAGL8 cDNA with a length of 711 bp was obtained through cloning using PCR amplification technology.Bioinformatics analysis results showed that MrAGL8 encoded 236 amino acids,it had MADS-box and K-box protein conserve domains.Its molecular weight was 27.39 ku,the theoretical isoelectric point was 8.69,the total number of positively charged residues was 39,the total number of negatively charged residues was 36,and the instability coefficient was 48.5.The secondary structure of the protein contained α-helix and β-sheet.It was an unstable protein and belonged to a hydrophilic alkaline protein.Subcellular localization results showed that MrAGL8 protein was located in the nucleus.The expression analysis of different tissues and organs showed that the expression level of MrAGL8 in different tissues was stem>root>pod>leaf>flower,and the expression levels of roots and stems were significantly different from those of leaves,flowers and pods.The results showed that MrAGL8 gene was related to pod dehiscence.

Key words: Medicago ruthenica, MrAGL8, Gene cloning, Subcellular localization, Expression analysis

CLC Number:

S541.9

YU Jia, GUO Huiqin, LI Yuxia, LEI Hui, REN Weibo. Cloning,Subcellular Localization and Expression Analysis of MrAGL8 Gene in Medicago ruthenica[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 55-61. doi: 10.7668/hbnxb.20195685.

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Figures/Tables 10

Fig.1 PCR amplification and electrophoresis detection of MrAGL8 CDS M.Trans 2K DNA Ladder;1—4.PCR产物。 M.Trans 2K DNA Ladder;1—4.PCR products.

Fig.2 PCR amplification and electrophoresis detection of MrAGL8 CDS colonies M.Trans 2K DNA Ladder;1—8.PCR 产物。 M.Trans 2K DNA Ladder;1—8.PCR products.

Fig.3 MrAGL8 cDNA coding region sequence and coded amino acid sequence

Fig.4 Domain prediction of MrAGL8

Fig.5 Phylogenetic tree of MrAGL8

Fig.6 Hydrophilic or hydrophobic analysis of MrAGL8

Fig.7 Secondary structure prediction of MrAGL8 protein Blue.α-helix;Green.β-turn;Purple.Extended strand;Orange.Random coil.

Fig.8 Tertiary structure prediction of MrAGL8 protein

Fig.9 Subcellular localization of MrAGL8 in lower epidermal cells of tobacco leaves Bright.Bright field;GFP.Green fluorescence channel;DAPI(4,6-diamidino-2-phenylindole). Positive control for nuclear localization;Merged.Mixed field

Fig.10 The relative expression of MrAGL8 gene in different tissues Different lowercase letters indicate significant differences in different treatments(P<0.05).

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