Cloning and Expression Analysis of Stress-induced-Phosphoprotein 1 (FoSTIP1) in Fusarium oxysporum f. sp. cubense race 4

doi:10.3969/j.issn.1000-2561.2019.02.019

Abstract

Abstract:

In this study, a putative target gene of FoSkn7, a transcription factor in Fusarium oxysporum f. sp. cubense race 4 (Foc4) was cloned. The result showed that the ORF sequence was 1737 nt and encoded a protein containing 578 amino acids. Domain and phylogenetic analysis showed that the protein contained a STI1 domain and several tetratricopeptide repeat (TPR) domains. The protein had the highest similarity with the stress-induced-phosphoprotein 1 of F. oxysporum f. sp. lycopersici 4287, indicating that the protein was STIP1 in Foc4 and named FoSTIP1. To analyze the expression of FoSTIP1 in Foc4 during invasion of the roots of banana seedling sorbeing induced by H₂O₂, the real time RT-PCR was performed. The results showed that the expression of FoSTIP1 was significantly up-regulated under these two conditions. The different expression of FoSTIP1 between wild-type B2 strain and FoSKN7 deletion mutant were also analyzed by the real time RT-PCR after being induced by H₂O₂. The result showed that the expression of FoSTIP1 in FoSKN7 deletion mutant was much lower than that of FoSTIP1 in B2 strain, even it was induced by H₂O₂. The results suggested that the putative target gene of FoSkn7 may be involved in the process of Foc4 resistance to exogenous oxidative stress.

Key words: Foc4, STIP1, expression characteristics

CLC Number:

S432.4

QI Xingzhu,WANG Jun,LIU Lei. Cloning and Expression Analysis of Stress-induced-Phosphoprotein 1 (FoSTIP1) in Fusarium oxysporum f. sp. cubense race 4[J]. Chinese Journal of Tropical Crops, 2019, 40(2): 341-347.

Figures/Tables 7

Tab. 1 Primers and their sequences in this study

引物名称 Primer name	引物序列Primer sequence (5°-3°)
5°-STpromt	GGCAACGGTTTCGTGATG
3°-STpromt	CCGCTGGGATCTTGAGGT
5°-STgemic	TGCATAGACTTGCATCGAATG
3°-STgemic	TCGCCATCATCTCCTTCG
5°-STorf	TAATTCACACACGAAATGGCTT
3°-STorf	GCCATCATCTCCTTCGCTTA
5°-STyg	TCCCTTACCGAGCACCGAACT
3°-STyg	TAGGCAGCAACGGCACCAG
5°- Actin	CCGTGACATCAAGGAGAAGC
3°- Actin	GGAAAGTGGACAGGGAAGCA

Fig. 1 Structural analysis of FoSTIP1 gene on genome The blue strip is the pre-acquired cDNA.

Fig. 2 Domain analysis of FoSTIP1. A: Several TPR (tetratricopeptide repeat) domain (from 258 to 495) in FoSTIP1; B: A typical STI1 (stress-inducible protein-1) domain (from 526 to 565) in the C terminal of FoSTIP1.

Fig. 3 The Phylogenetic tree of FoSTIP1 of Foc4 and its homologous The FoSTIP1 was marked by ●. The homologous protein sequences used in the phylogenetic analysis were derived from NCBI.

Fig. 4 Relative transcripts abundance of FoSTIP1 of wild-type strain B2 in oxidative stress of H2O2 for 5 h or being induced by banana tissue seedlings for 24 h were examined Polymerase chain reaction assays were carried out in triplicate.

Fig. 5 Relative transcripts abundance of FoSTIP1 of wild-type strain B2 and FoSNK7 deletion mutant in oxidative stress of H2O2 for 5 h Polymerase chain reaction assays were carried out in triplicate.

Fig. 6 The promoter sequence of FoSTIP1 The putative Skn7 binding sites were marked by _ , and “ATG ” was the starting codon of ORF of FoSTIP1.

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