松材线虫性别决定基因Bx-sex-1表达特征和生物学功能分析

韦鹏飞; 理永霞; 冯宇倩; 刘振凯; 张星耀

doi:10.12302/j.issn.1000-2006.202104004

松材线虫性别决定基因Bx-sex-1表达特征和生物学功能分析

韦鹏飞, 理永霞, 冯宇倩, 刘振凯, 张星耀

南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4) : 15-20.

PDF(1679 KB)

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

PDF(1679 KB)

南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4) : 15-20. DOI: 10.12302/j.issn.1000-2006.202104004

专题报道Ⅰ：松材线虫病致病机理与病害防治（执行主编　叶建仁）

松材线虫性别决定基因Bx-sex-1表达特征和生物学功能分析

韦鹏飞 ¹^,² ,
理永霞 ¹^,²^,^* ,
冯宇倩 ¹^,² ,
刘振凯 ¹^,² ,
张星耀 ¹^,²

作者信息 +

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

WEI Pengfei ¹^,² ,
LI Yongxia ¹^,²^,^* ,
FENG Yuqian ¹^,² ,
LIU Zhenkai ¹^,² ,
ZHANG Xingyao ¹^,²

Author information +

文章历史 +

摘要

【目的】揭示松材线虫(Bursaphelenchus xylophilus)性别决定基因Bx-sex-1的表达特征和生物学功能。【方法】通过Mega 6中的Jones-Taylor-Thornton (JTT)模型,采用邻接法(neighbor joining,NJ)对Bx-sex-1和其他线虫的氨基酸序列进行系统发育构建。采用同步培养的方法分别收集不同龄期的松材线虫并提取总RNA,反转录获得cDNA并采用实时荧光定量PCR (qRT-PCR) 的方法,研究Bx-sex-1在松材线虫不同发育阶段的相对表达量。同时采用qRT-PCR并通过绝对定量法分析Bx-sex-1基因在雌虫和雄虫基因组的拷贝数。采用dsRNA浸泡法对目的基因进行干扰,观察松材线虫后代卵的孵化率和雌雄比的变化。【结果】系统进化树显示松材线虫Bx-sex-1和其他两个植物寄生线虫成为一支,与自由生线虫和寄生动物线虫分别进化为不同的分支。松材线虫Bx-sex-1在不同发育阶段相对表达量有差异,在卵期相对表达量最高,而在2龄幼虫时期最低,从2龄幼虫时期到成虫时期相对表达量逐渐升高。qRT-PCR结果显示Bx-sex-1基因在雌虫和雄虫基因组中均为单拷贝。Bx-sex-1被干扰后,卵的孵化率下降,雌雄比降低。【结论】松材线虫Bx-sex-1基因在卵的发育过程和性别决定中可能具有重要作用。

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.

导出引用

韦鹏飞, 理永霞, 冯宇倩, 等. 松材线虫性别决定基因Bx-sex-1表达特征和生物学功能分析[J]. 南京林业大学学报（自然科学版）. 2022, 46(4): 15-20 https://doi.org/10.12302/j.issn.1000-2006.202104004

WEI Pengfei, LI Yongxia, FENG Yuqian, et al. 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 https://doi.org/10.12302/j.issn.1000-2006.202104004

中图分类号： S763.11

参考文献

列表( 原文顺序 | 文献年度倒序 | 文中引用次数倒序 ) 可视化分析

[1]	YI C K, BYUN B H, PARK J D, et al. First finding of the pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle and its insect vector in Korea[J]. Research Reports of the Forestry Research Institute, 1989(38): 141-149. 本文引用 [1]

[2]	FUTAI K. Pine wilt in Japan: from first incidence to the present[C]// ZHAO B G, FUTAI K, SUTHERLAND J R, et al. Pine wilt disease. Japan: Springer, 2008:5-12. DOI: 10.1007/978-4-431-75655-2_2. 本文引用 [1]

[3]

丁晓磊, 张悦, 林司曦, 等. 基于高通量测序技术的松材线虫研究进展[J]. 南京林业大学学报(自然科学版), 2022, 46(4):1-7.

DING

X L

, ZHANG

, LIN

S X

, et al. An overview of high-throughput sequencing techniques applied on Bursaphelenchus xylophilus[J]. J Nanjing For Univ(Nat Sci Ed): 2022, 46(4):1-7.DOI:10.12302/j.issn.1000-2006.202204039.

本文引用 [1]

[4]	KIKUCHI T, AIKAWA T. Estimation of virulence of Bursaphelenchus xylophilus (Nematoda:Aphelenchoididae) based on its reproductive ability[J]. Nematology, 2007, 9(3): 371-377. DOI:10.1163/156854107781352007. 本文引用 [2]

[5]	HASEGAWA K, MOTA M M, FUTAI K, et al. Chromosome structure and behaviour in Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) germ cells and early embryo[J]. Nematology, 2006, 8(3): 425-434. DOI:10.1163/156854106778493475. 本文引用 [3]

[6]	HASEGAWA K, MOTA M, FUTAI K, et al. Developmental biology and cytogenetics of Bursaphelenchus xylophilus[G]//Pine wilt disease: a worldwide threat to forest ecosystems. Dordrecht:Springer, 2008: 91-100.DOI:10.1007/978-1-4020-8455-3_8. 本文引用 [1]

[7]	KIYOHARA T. Sexual attraction in Bursaphelenchus xylophilus[J]. Jpn J Nematol, 1982, 11:7-12.DOI:10.14855/jjn1972.11.7. 本文引用 [1]

[8]	GAO M G, LI Y X, ZHANG W, et al. Bx-daf-22 contributes to mate attraction in the gonochoristic nematode Bursaphelenchus xylophilus[J]. Int J Mol Sci, 2019, 20(17): 4316.DOI:10.3390/ijms20174316. 本文引用 [1]

[9]	BRIDGES C B. Non-disjunction as proof of the chromosome theory of heredity (concluded)[J]. Genetics, 1916, 1(2): 107-163. DOI:10.1093/genetics/1.2.107. 本文引用 [1]

[10]	MADL J E, HERMAN R K. Polyploids and sex determination in Caenorhabditis elegans[J]. Genetics, 1979, 93(2): 393-402.DOI:10.1093/genetics/93.2.393. 本文引用 [1]

[11]	GLADDEN J M, FARBOUD B, MEYER B J. Revisiting the X:A signal that specifies Caenorhabditis elegans sexual fate[J]. Genetics, 2007, 177(3):1639-1654. DOI:10.1534/genetics.107.078071. 本文引用 [3]

[12]	CARMI I, KOPCZYNSKI J B, MEYER B J. The nuclear hormone receptor SEX-1 is an X-chromosome signal that determines nematode sex[J]. Nature, 1998, 396(6707):168-173.DOI:10.1038/24164. 本文引用 [3]

[13]	ZHOU L F, MA X X, ZHU N J, et al. The role of Mab-3 in spermatogenesis and ontogenesis of pinewood nematode, Bursaphelenchus xylophilus[J]. Pest Manag Sci, 2021, 77(1):138-147.DOI:10.1002/ps.6001. 本文引用 [1]

[14]	李江梅. 松材线虫tra-1基因克隆及功能分析[D]. 泰安: 山东农业大学, 2019. LI J M. Cloning and function analysis of tra-1 gene of Bursaphelenchus xylophilus[D]. Tai’an: Shandong Agricultural University, 2019. 本文引用 [1]

[15]	HIDEAKI I, KAZUYOSHI F. A simple method for disinfection of pine wood nematode eggs using their adhesiveness[J]. Jpn J Nematol, 1985, 15: 64.DOI:10.14855/jjn1972.15.64. 本文引用 [1]

[16]	FUTAI K, TAKEUCHI Y, SHINYA R. A technique for separating the developmental stages of the propagative form of the pine wood nematode, Bursaphelenchus xylophilus[J]. Nematology, 2009, 11(2):305-307.DOI:10.1163/156854108x399164. 本文引用 [1]

[17]	牛玉娜. 五指山猪KIT基因拷贝数检测暨猪PSME1基因与免疫性状关系的研究[D]. 武汉: 华中农业大学, 2007. NIU Y N. The copy number detection of KIT gene in Wuzhishan pig and association analysis between porcine PSME1 gene and immunity traits[D]. Wuhan: Huazhong Agricultural University, 2007. 本文引用 [1]

[18]	王殿东. 松材线虫休眠相关基因daf-3, daf-9, daf-12, daf-16的克隆及其功能研究[D]. 长春: 吉林大学, 2012. WANG D D. Cloning and function research of dormancy related gene daf-3, daf-9, daf-12, daf-16 in pine wood nematode, Bursaphelenchus xylophilus[D]. Changchun: Jilin University, 2012. 本文引用 [1]

[19]	MAMIYA Y. Population increase of Bursaphelenchus lignicolus in wood of pine seedlings in relation with the time after inoculation[J]. Forestry Society, 1974: 249-251. 本文引用 [1]

[20]	HAAG E S. The evolution of nematode sex determination: C. elegans as a reference point for comparative biology[C]// The C. elegans Research Community. Worm book, 2005. 本文引用 [1]

[21]	REECE-HOYES J S, SHINGLES J, DUPUY D, et al. Insight into transcription factor gene duplication from Caenorhabditis elegans promoterome-driven expression patterns[J]. BMC Genomics, 2007, 8: 27.DOI:10.1186/1471-2164-8-27. 本文引用 [1]

[22]	HARGITAI B, KUTNYÁNSZKY V, BLAUWKAMP T A, et al. Xol-1, the master sex-switch gene in C.elegans, is a transcriptional target of the terminal sex-determining factor TRA-1[J]. Development, 2009, 136(23): 3881-3887. DOI:10.1242/dev.034637. 本文引用 [1]

[23]	CSANKOVSZKI G, COLLETTE K, SPAHL K, et al. Three distinct condensin complexes control C.elegans chromosome dynamics[J]. Curr Biol, 2009, 19(1):9-19.DOI:10.1016/j.cub.2008.12.006. 本文引用 [1]

[24]	LEHNER B, CALIXTO A, CROMBIE C, et al. Loss of LIN-35, the Caenorhabditis elegans ortholog of the tumor suppressor p105Rb, results in enhanced RNA interference[J]. Genome Biol, 2006, 7(1): R4. DOI:10.1186/gb-2006-7-1-r4. 本文引用 [1]

[25]	HAAG E S, WANG S P, KIMBLE J. Rapid coevolution of the nematode sex-determining genes fem-3 and tra-2[J]. Curr Biol, 2002, 12(23):2035-2041. DOI:10.1016/s0960-9822(02)01333-7. 本文引用 [1]

[26]	HAAG E S, KIMBLE J. Regulatory elements required for development of Caenorhabditis elegans hermaphrodites are conserved in the tra-2 homologue of C. remanei, a male/female sister species[J]. Genetics, 2000, 155(1): 105-116.DOI:10.1093/genetics/155.1.105. 本文引用 [1]

[27]	KIKUCHI T, COTTON J A, DALZELL J J, et al. Genomic insights into the origin of parasitism in the emerging plant pathogen Bursaphelenchus xylophilus[J]. PLoS Pathog, 2011, 7(9): e1002219.DOI:10.1371/journal.ppat.1002219. 本文引用 [1]