Expression characteristics and biological functions of the sex determination gene Bx-sex-1 in Bursaphelenchus xylophilus

doi:10.12302/j.issn.1000-2006.202104004

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (4): 15-20.doi: 10.12302/j.issn.1000-2006.202104004

Special Issue: 松材线虫病致病机理与病害防治

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Expression characteristics and biological functions of the sex determination gene Bx-sex-1 in Bursaphelenchus xylophilus

WEI Pengfei¹^,²(), LI Yongxia¹^,²^,^*(), FENG Yuqian¹^,², LIU Zhenkai¹^,², ZHANG Xingyao¹^,²

1. Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
2. Key Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, China

Received:2021-04-02 Revised:2021-08-01 Online:2022-07-30 Published:2022-08-01
Contact: LI Yongxia E-mail:ftzwpf@126.com;liyongxiaxjs@163.com

Abstract

Abstract:

【Objective】 This study aimed to determine the expression characteristics and biological functions of Bx-sex-1, a sex determination gene in Bursaphelenchus xylophilus. 【Method】 The phylogenetic analysis was performed using the neighbor joining (NJ) method, based on the Jones-Taylor-Thornton (JTT) model in the molecular evolutionary genetic analysis (Mega 6). After collecting pine wood nematodes at different developmental stages using synchronous culture, cDNA was synthesized from the extracted RNA, and the relative expression level of Bx-sex-1 was studied using real-time quantitative PCR (qRT-PCR), where the copy number of Bx-sex-1 in the female and male genomes was analyzed through the absolute quantitative analysis. B. xylophilus adults were socked with double-strand RNA (dsRNA), and the hatching rate and the female to male sex ratio of the progeny were observed. 【Result】 The phylogenetic tree showed that the pine wood nematode Bx-sex-1 was a clade with two other plant parasitic nematodes, and had evolved into a different clade from the free and animal parasitic nematodes, respectively. The Bx-sex-1 gene exhibited the highest relative expression level in the egg period, with the lowest expression in the L₂ period that increased gradually up to the adult period. Both female and male genomes had a single copy of the gene. Compared with untreated B. xylophilus, the hatching rate of the eggs and the sex ratio decreased in the experimental samples. 【Conclusion】 The Bx-sex-1 gene of B. xylophilus may play an important role in egg development and sex determination.

Key words: Bursaphelenchus xylophilus, sex determination gene, RNA interference(RNAi), sex ratio of female to male

CLC Number:

S763.11

WEI Pengfei, LI Yongxia, FENG Yuqian, LIU Zhenkai, ZHANG Xingyao. Expression characteristics and biological functions of the sex determination gene Bx-sex-1 in Bursaphelenchus xylophilus[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(4): 15-20.

Figures/Tables 5

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References 27

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引物名称 primer name	序列(5'—3') sequence(5'—3')
Bx-sex-1-5'-F	ATGGTCATGGATAATGTAGGC
Bx-sex-1-5'-R	ACAACTCTGACAACGAACT
Bx-sex-1-3'-F	AAACGGACCCTGAAGTCCCC
Bx-sex-1-3'-R	TCATGTCGCAGCCTGTCCTGATC
C-Bx-daf-12-F	TACGGTGGTCAACACGACTG
C-Bx-daf-12-R	TCGACCGGTTGACTCACAAG
C-Bx-sex-1-F	CAAAGGAGAACCGGACCGAA
C-Bx-sex-1-R	CTGGCCATTGGGGAGTGAAT
Bx-sex-1-T7-F	GGATCCTAATACGACTCACTATAGGGCTCCACAACCTCAACT
Bx-sex-1-R	GGCTCTGCTTGTTCATCA
Bx-sex-1-T7-R	GGATCCTAATACGACTCACTATAGGGGCTCTGCTTGTTCATCA
Bx-sex-1-F	GCTCCATCAACCTCAACT
q-Bx-sex-1-F	CTTCAAGGAGTCGGAATAATG
q-Bx-sex-1-R	ACTGGAGTCGTATGATTGG
β-actin-F	TCCGTACCCTGAAGTTGGCTAACC
β-actin-R	AAGTGGAGACGAGGGAATGGAACC

样本 sample	拷贝数/×10^-6 copy number		Bx-daf-12与Bx-sex-1 拷贝数之比 copy number ratio of Bx-daf-12 to Bx-sex-1
样本 sample	Bx-daf-12	Bx-sex-1
雌虫1 female 1	6.33	5.10	1.24
雌虫2 female 2	4.87	3.74	1.30
雄虫1 male 1	3.80	3.01	1.26
雄虫2 male 2	3.04	2.31	1.31

Expression characteristics and biological functions of the sex determination gene Bx-sex-1 in Bursaphelenchus xylophilus

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References 27

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