Cloning and Expression Patterns of StRac5,StRac7 and StRac13 Genes in Potato

doi:10.7668/hbnxb.20194582

Abstract

Abstract:

To explore the expression patterns of potato genes and provide theoretical basis for the subsequent study of gene resistance to potato diseases.Three endogenous small G protein genes StRac5,StRac7 and StRac13 were cloned from potato,and their bioinformatics and expression patterns were analyzed.The results showed that StRac5,StRac7 and StRac13 contained similar conserved sequences to the small G proteins in Arabidopsis and rice,all belonging to ROP proteins.The protein domains of StRac5 and StRac13 were identical with AtRop1,AtRop3,AtRop5,AtRop6 of Arabidopsis and rice OsRac5,OsRac6,OsRac7,and StRac7 was identical with AtRop9.Phylogenetic tree analysis showed that StRac7 belonged to class Ⅱ and was closest to Arabidopsis AtRop9.StRac13 belonged to class Ⅲ and was the closest genetic relative to Arabidopsis AtRop7.StRac5 belonged to class IV and was the closest genetic relative to rice OsRac5 and OsRac6.The relative expression levels of StRac5,StRac7 and StRac13 genes in different tissues of potato were leaf>root>stem.Compared with the expression in stems,the expression of StRac5,StRac7 and StRac13 in leaves were increased by 1 222.4%,2 531.3% and 468.2%,respectively.After inoculation,the relative expression of StRac5,StRac7 and StRac13 genes increased first and then decreased,and the relative expression of StRac5 and StRac13 genes increased by 62.2%,40.4%,respectively,compared with 0 h after inoculation 24 h.The relative expression of StRac7 gene was increased by 827.8% at 72 h after inoculation compared with 0 h.After ABA,SA,GA and 6-BA treatment,the relative expression of StRac5,StRac7 and StRac13 genes showed a downward trend.After JA and IAA treatment,the relative expression of StRac5,StRac7 and StRac13 genes showed an up-regulated trend.Therefore,StRac5,StRac7 and StRac13 genes can respond to potato late blight infection,and their expression patterns are different in different tissues and under different hormone treatments.

Key words: Potato, Small G protein, Gene cloning, Late blight, Expression pattern

GAO Wanting, LIU Zhida, SUN Xuetao, LI Zhiping, LÜ Wenxia, LI Aizhen, ZHAO Jun, ZHANG Zhiwei. Cloning and Expression Patterns of StRac5,StRac7 and StRac13 Genes in Potato[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(5): 33-41. doi: 10.7668/hbnxb.20194582.

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Figures/Tables 11

Tab.1 Information of primer sequences

引物名称 Primer name	引物序列(5'-3') Primer sequence(5'-3')	引物用途 Purpose of primer
StRac5-F	ATGAGTGCTTCTAGGTTTATAAAGTG	基因克隆
StRac5-R	TCACAAGATAGAGCAGGCTTTC
StRac7-F	ATGAATGCTTCAAAGTTCATCAAATG
StRac7-R	CTAAGCAACACAGCCTCCACA
StRac13-F	ATGACTACTGCTAGATTCATCAAATG
StRac13-R	TCAAAGTATTGCACATAACGTCCTT
qStActin-F	CACCCTGTTCTGCTCACT	表达量分析
qStActin-R	CAGCCTGAATAGCAACATAC
qStRac5-F	GGACCATCCAGGTGCTGTTC
qStRac5-R	GCGGCATCAAAAACAGCCTTA
qStRac7-F	TTCAGTGCCAATGTAGCCGT
qStRac7-R	CGACGAAGTTCAGGCATCCA
qStRac13-F	ATGTGGTGGTGGATGGTAGC
qStRac13-R	TGTTCTCATAGCTTGCCTTGCT

Fig.1 Cloning of StRac5,StRac7,and StRac13 genes M.DM2000 DNA Marker。

Fig.2 cDNA sequence of StRac5 gene and its encoded amino acid sequence

Fig.3 cDNA sequence of StRac7 gene and its encoded amino acid sequence

Fig.4 cDNA sequence of StRac13 gene and its encoded amino acid sequence

Fig.5 Comparison of conserved sequences of StRac5,StRac7 and StRac13 proteins

Fig.6 Distribution of protein conserved domains(Motifs) of StRac5,StRac7 and StRac13 in potato

Fig.7 Phylogenetic tree of StRac5,StRac7 and StRac13 proteins in potato

Fig.8 Relative expression of StRac5,StRac7 and StRac13 genes in different tissues of potato Different lowercase letters represent significant difference in different treatment(P<0.05).The same as Fig.9-10.

Fig.9 Relative expression of StRac5,StRac7 and StRac13 genes after inoculation with Phytophthora latella

Fig.10 Relative expression of StRac5,StRac7 and StRac13 genes after exogenous hormone treatment

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