Cloning and Functional Analysis of FaGH3.17 Gene in Strawberry

doi:10.16420/j.issn.0513-353x.2023-1008

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (11): 2483-2494.doi: 10.16420/j.issn.0513-353x.2023-1008

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Next Articles

Cloning and Functional Analysis of FaGH3.17 Gene in Strawberry

YANG Juanbo, GUO Lili, LU Shixiong, GOU Huimin, WANG Shuaiting, ZENG Baozhen, MAO Juan^*()

Collage of Horticulture，Gansu Agricultural University，Lanzhou 730000，China

Received:2024-07-18 Revised:2024-08-29 Online:2024-12-12 Published:2024-11-25
Contact: MAO Juan

Abstract

Abstract:

In order to study the response of GH3.17 gene to salt stress in strawberry，the evolutionary relationship and physicochemical properties of FaGH3.17 were analysed using bioinformatics，and the gene was cloned and subsequently subcellularly localised in tobacco and heterologously overexpressed in Arabidopsis to verify its function in salt stress. The results showed that FaGH3.17 is an acidic and unstable hydrophobic，non-secretory protein，and the analysis of the evolutionary relationship of species revealed that FaGH3.17 is the closest relative to Potentilla anserina. Subcellular localisation in tobacco revealed that FaGH3.17 was mainly localised in the nucleus and cell membrane. Heterologously overexpression analysis in Arabidopsis thaliana revealed that under salt stress，the conductivity，MDA and H₂O₂ content were higher than those of wild-type plants，which were elevated by 15.43%，42.97%，and 18.86%，respectively，whereas the Pro content，POD，SOD，and CAT activities were significantly lower than those of the wild-type plants，which were reduced by 13.28%，15.42%，14.22%，19.87%，respectively. And the relative expression of salt-responsive genes were all significantly reduced. It was shown that heterologous overexpression of FaGH3.17 significantly reduced the antioxidant enzyme activities and osmoregulatory substance contents in Arabidopsis，which predicts that GH3.17 may attenuate the salt tolerance of plants.

Key words: strawberry, gene cloning, salt stress, functional verification

YANG Juanbo, GUO Lili, LU Shixiong, GOU Huimin, WANG Shuaiting, ZENG Baozhen, MAO Juan. Cloning and Functional Analysis of FaGH3.17 Gene in Strawberry[J]. Acta Horticulturae Sinica, 2024, 51(11): 2483-2494.

Figures/Tables 9

Table 1 Sequence of real-time fluorescence quantitative primers

基因 Gene	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
FaGH3.17	ACAGTTGATCTGGTGGATGTGAAGC	ACGGAACTGAGGAGCATTGTTGTG
AtHKT1	GTCTCTGCCATCACCGTCTCTTC	GCCACCGAGGAACATGAGGATAG
AtSOS1	TCATCTCCCGCCGCATTATCAC	GATTGCTCTTGCTCTCGTCTCAAC
AtNHX1	TTGACAAGTGGAGATCCGTGAGTG	TGCTCTTCCAACCATGACCAGAC
AtAPX1	CTCTGGGACGATGCCACAAG	CTCGACCAAAGGACGGAAAA
AtActin2	CTGGCCTACGTGGCACTTGACTT	AGCGATGGCTGGAACAGAAC

Fig. 1 PCR detection of FaGH3.17 gene in strawberry（A）and subcellular mapping in tobacco（B）

Fig. 2 Phylogenetic analysis of FaGH3.17 with proteins from other species

Fig. 3 Motif analysis of FaGH3.17 protein

Fig. 4 Identification of FaGH3.17 heterologously overexpressing Arabidopsis thaliana（A）and phenotypic observation of overexpressing Arabidopsis thaliana under salt stress 15 days（B） M：DL2000 marker；+：Positive control；-：Negative control；WT：Wild type；OE：Heterologous overexpression strain. The same below.

Fig. 5 Analysis of FaGH3.17 heterologously expression（A）and hydrogen peroxide content（B）in overexpressing Arabidopsis thaliana after salt stress 15 days Lowercase letters represent significance level at P < 0.05. The same below.

Fig. 6 Analysis of cytoplasmic membrane permeability under salt stress in FaGH3.17 heterologously overexpressing Arabidopsis thaliana

Fig. 7 Determination of antioxidant enzyme activities of FaGH3.17 heterologously overexpressing Arabidopsis thaliana under salt stress

Fig. 8 Expression levels of 4 salt-responsive genes under salt in Arabidopsis

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Cloning and Functional Analysis of FaGH3.17 Gene in Strawberry

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