A Primary Study on FvMOB1 Interact with FvM4K1 and Response to Auxin Regulation in Fragraria vesca

doi:10.16420/j.issn.0513-353x.2020-0859

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (11): 2185-2196.doi: 10.16420/j.issn.0513-353x.2020-0859

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A Primary Study on FvMOB1 Interact with FvM4K1 and Response to Auxin Regulation in Fragraria vesca

LIU Lu, GU Si, ZHANG Weiwei, ZHAO Shuaiqi, LI Anran, XING Yu()

Advanced Innovation Center for Forest Molecular Design and Breeding,Beijing Key Laboratory for Agricultural Application and New Technique,College of Plant Science and Technology,Beijing University of Agriculture,Beijing 102206,China

Received:2020-12-17 Revised:2021-03-05 Published:2021-12-02
Contact: XING Yu E-mail:xingyu@bua.edu.cn

Abstract

Abstract:

In order to study how FvMOB1 genes involved in auxin mediated woodland strawberry growth and regulate cell proliferation,this experiment using woodland strawberry（Fragraria vesca）‘Hawaii 4’material,cloned and analyzed coding sequence of homologous gene FvMOB1A and FvMOB1B based on bioinformatics. Fluorescence quantitative PCR was applied to analysis FvMOB1’s relative expression quantity in different organs of woodland strawberry,different development periods of fruit and different days during IAA treatment. Carried out subcellular localization of FvMOB1 and yeast two-hybrid interaction. The results show that FvMOB1A and FvMOB1B encode 215 and 216 amino acids respectively with open reading frame of 648 and 651 bp. In woodland strawberry,the expression level of FvMOB1A was much higher than that of FvMOB1B,the expression level of FvMOB1A is high in achene and root and lowest in stem,while the expression level of FvMOB1B is relatively highest in achene and lowest in leaves. The expression of both FvMOB1A and FvMOB1B are highest in the 30th day after flowering （post ripening stage）. The expression patterns of the two genes are the same after IAA treatment,both are up-regulated and reaching the peak value on the 7th day. The localization of FvMOB1A and FvMOB1B are in the cell nucleus and cell membrane,and yeast two-hybrid verified that both FvMOB1A and FvMOB1B can interact with mitogen-activated protein kinase FvM4K1.

Key words: woodland strawberry, MOB1, auxin, cell proliferation

CLC Number:

S668.4

LIU Lu, GU Si, ZHANG Weiwei, ZHAO Shuaiqi, LI Anran, XING Yu. A Primary Study on FvMOB1 Interact with FvM4K1 and Response to Auxin Regulation in Fragraria vesca[J]. Acta Horticulturae Sinica, 2021, 48(11): 2185-2196.

Figures/Tables 9

Table 1 Primers used in this study

引物名称 Primer name	序列（5′-3′） Sequence
FvMOB1A-F	ATGAGTCTCTTCGGTCTGGG
FvMOB1A-R	GGGTCAATAAGGAACGATGATAGA
FvMOB1B-F	ATGAGTCTCTTTGGTCTTGGAAG
FvMOB1B-R	TCACTTTGGCTGTAGAATAGAGTC
FvMOB1A-QF	TTGATGGACTGGATTGAAT
FvMOB1A-QR	TGAAGCAGGTATTTAGATGG
FvMOB1B-QF	GCAGGACCAAAGTATGAG
FvMOB1B-QR	CACGGAACATTCGCTTAA
FvActin-QF	GGGCCAGAAAGATGCTTATGTCGG
FvActin-QR	GGGCAACACGAAGCTCATTGTAGAAG
FvMOB1A-LF	ATCGACTCTAGAAAGCTTATGAGTCTCTTCGGTCTG
FvMOB1A-LR	CTCCTCGCCCTTGCTCACCATGGTACCATAAGGAACGATGATAGATTCT
FvMOB1B-LF	ATCGACTCTAGAAAGCTTATGAGTCTCTTTGGTCTT
FvMOB1B-LR	CTCCTCGCCCTTGCTCACCATGGTACCCTTTGGCTGTAGAATAGAGTC
FvMOB1A-AF	ATATGGCCATGGAGGCCAGTGAATTCATGAGTCTCTTCGGTCTGG
FvMOB1A-AR	CGATTCATCTGCAGCTCGAGTCAATAAGGAACGATGATAGATTCT
FvMOB1B-AF	ATATGGCCATGGAGGCCAGTGAATTCATGAGTCTCTTTGGTCTTGGAAG
FvMOB1B-AR	TATCTACGATTCATCTGCAGCTCGAGTCACTTTGGCTGTAGAATAGAGTCA
FvM4K1-BF	TGCATATGGCCATGGAGGCCAGTGAATTCATGGCTTTCTTATTCTTCCT
FvM4K1-BR	TGCGGCCGCTGCAGGTCGACGGATCCCTACAAACGAAGAATGGTTCTTA

Fig. 1 The PCR result of FvMOB1A and FvMOB1B genes

Fig. 2 Amino acid sequence alignment of MOB1 in different species Sc:Saccharomyces cerevisiae;At:Arabidopsis thaliana;Fv:Fragraria vesca;Ms:Medicago sativa; Dm:Drosophila melanogaster;Hs:Homo sapiens. Frame means Mob1/phocein family domain.

Fig. 3 Phylogenetic tree of MOB1 gene based on amino acid

Fig. 4 Relative expression of FvMOB1 in different organs Different lowercase letters indicate significant differences（P < 0.05）.

Fig. 5 Relative expression of FvMOB1 in different development stages of fruit Different lowercase letters indicate significant differences（P < 0.05）.

Fig. 6 Relative expression of FvMOB1 in seedlings of woodland strawberry after IAA treatment Different lowercase letters indicate significant differences（P < 0.05）.

Fig. 7 Subcellular localization of protein FvMOB1A and FvMOB1B

Fig. 8 Yeast two-hybrid verification the interaction of FvMOB1 and FvM4K1

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A Primary Study on FvMOB1 Interact with FvM4K1 and Response to Auxin Regulation in Fragraria vesca

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