Transcriptional activity of the Bx-HSF-1 in Bursaphelenchus xylophilus (Aphelenchida: Aphelenchoididae)

ZHANG Ruizhi, JIANG Shengwei, WU Hao, CHEN Qiaoli, LI Danlei, WANG Feng

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (4) : 8-14.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (4) : 8-14. DOI: 10.12302/j.issn.1000-2006.202102019

Transcriptional activity of the Bx-HSF-1 in Bursaphelenchus xylophilus (Aphelenchida: Aphelenchoididae)

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Abstract

【Objective】 The transcriptional activity of Bx-HSF-1 and the expression of its target genes at the seven life stages of Bursaphelenchus xylophilus were studied to reveal its role in the growth and development of B. xylophilus. 【Method】 The CDS (coding sequence) of Bx-hsf-1 gene was constructed into plasmid pGBKT7 to obtain the yeast (Saccharomyces cerevisiae) bait expression vector pGBKT7-Bx-hsf-1. The self-activation of pGBKT7-Bx-hsf-1 was detected. RNA interference and transcriptome sequencing techniques were used to identify the differentially expressed genes of B. xylophilus after Bx-hsf-1 gene was knocked down. The gene interaction analysis of differentially expressed genes was analyzed, and the target genes that were directly regulated by Bx-HSF-1 were screened out. RT-qPCR (quantitative real-time PCR) was used to identify the expression levels of five genes in J2, DJ3, DJ4, J3, J4, female, and male stages of B. xylophilus. 【Result】 The pGBKT7-Bx-hsf-1 yeast turned blue on SD/-Trp/-His/-Ade+X-α-gal medium and could grow on SD/-Trp and SD/-Trp/-His/-Ade mediums. A yeast transactivation activity assay suggested that Bx-HSF-1 had transcriptional activity. RT-qPCR results revealed the significant suppression of Bx-hsf-1 after RNAi treatment for 12 h. The total 110 differentially expressed genes were identified using a transcriptome analysis, of which 86 were down regulated. Total five target genes (Bx-daf-21, Bx-hsp-1, Bx-hsp-70, Bx-sti and Bx-cdc) were filtered using a gene interaction analysis. The expression of Bx-daf-21 was consistent with the expression of Bx-hsf-1 in the development of seven life stages of B. xylophilus, whereas the expression of Bx-cdc in DJ3 and the expression of Bx-hsp-70 were inconsistent with the expression of Bx-hsf-1. The expression of Bx-hsp-1 and Bx-sti was not correlated with the expression of Bx-hsf-1 in the seven life stages. 【Conclusion】 Bx-HSF-1 had a transcriptional activation function and participated in the development process of B. xylophilus through transcriptional regulations.

Key words

Bursaphelenchus xylophilus / heat shock transcription factor / transcriptional activation / life stage

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ZHANG Ruizhi , JIANG Shengwei , WU Hao , et al . Transcriptional activity of the Bx-HSF-1 in Bursaphelenchus xylophilus (Aphelenchida: Aphelenchoididae)[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2022, 46(4): 8-14 https://doi.org/10.12302/j.issn.1000-2006.202102019

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