拟松材线虫组织蛋白酶基因的RNA干扰及功能研究

doi:10.3969/j.issn.1000-2006.201612052

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

南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (05): 65-70.doi: 10.3969/j.issn.1000-2006.201612052

拟松材线虫组织蛋白酶基因的RNA干扰及功能研究

潘妍毅^1,2,黄麟¹,吴小芹^1*

1.南京林业大学,南方现代林业协同创新中心,江苏省有害生物入侵预防与控制重点实验室,南京林业大学林学院,江苏南京 210037; 2. 淮安海关,江苏淮安 223001

出版日期:2018-09-15 发布日期:2018-09-15
基金资助:
收稿日期:2016-12-31 修回日期:2018-02-10 基金项目:江苏省农业科技自主创新资助项目(CX(16)1005); 江苏高校优势学科建设工程资助项目(PAPD) 第一作者:潘妍毅(panyy1@jsciq.gov.cn)。*通信作者:吴小芹(xqwu@njfu.edu.cn),教授。

Study on function of cathepsin bmcath1 in Bursaphelenchus mucronatus using method of RNA interference

PAN Yanyi^1,2, HUANG Lin¹, WU Xiaoqin^1*

1. Co-Innovation Center for the Sustainable Forestry in Southern China, Jiangsu Key Laboratory for Prevention and Management of Invasive Species, College of Forestry, Nanjing Forestry University, Nanjing 210037, China; 2. Huai'an Customs, Huai'an 223001,

Online:2018-09-15 Published:2018-09-15

摘要/Abstract

摘要： 【目的】揭示拟松材线虫组织蛋白酶基因bmcath1生物学功能。【方法】采用dsRNA浸泡法对靶标基因进行基因干扰,观察拟松材线虫在活体和离体情况下的繁殖力、取食速率、性别比例、体积及致病力的变化情况。【结果】组织蛋白酶基因bmcath1被干扰后,拟松材线虫繁殖力增强约75%,取食速率变快,致病力变强,并且拟松材线虫群体的雄、雌成虫比由3:5变为6:13,成虫所占的比例由8%提高至19%。【结论】组织蛋白酶基因bmcath1对拟松材线虫的繁殖力和致病力具有显著调节作用。

Abstract: 【Objective】This study aimed to analyze the function of cathepsin bmcath1 in Bursaphelenchus mucronatus.【Method】 Nematodes of B. mucronatus were treated with double-strand RNA(dsRNA)of bmcath1. Reproduction ability, feeding speed, sex ratio, body size, and pathogenicity were analyzed.【Result】Compared with the untreated B. mucronatus, the reproduction ability of nematodes treated by dsRNA of bmcath1 was increased by 75%. The dsRNA-treated nematodes also displayed a quicker feeding speed and stronger pathogenicity. The sex ratio of dsRNA-treated nematodes was changed from ♂3:♀5 to ♂6:♀13, and the percentage of adult nematodes increased from 8% to 19%.【Conclusion】Cathepsin bmcath1 has a significant effect on reproduction ability and pathogenicity in B. mucronatus.

中图分类号:

S763.1

潘妍毅,黄麟,吴小芹. 拟松材线虫组织蛋白酶基因的RNA干扰及功能研究[J]. 南京林业大学学报（自然科学版）, 2018, 42(05): 65-70.

PAN Yanyi, HUANG Lin, WU Xiaoqin. Study on function of cathepsin bmcath1 in Bursaphelenchus mucronatus using method of RNA interference[J].Journal of Nanjing Forestry University (Natural Science Edition), 2018, 42(05): 65-70.DOI: 10.3969/j.issn.1000-2006.201612052.

参考文献

[1] 董锦艳,李铷,张克勤.松材线虫生物防治研究进展[J].植物保护, 2005, 31(5): 9-15.DOI:10.3969/j.issn.0529-1542.2005.05.002.
DONG J Y, LI R, ZHANG K Q. Advances in biological control of Bursaphelenchus xylophilus by natural enemies and nematocidal compounds from plants[J]. Plant Protection, 2005, 31(5): 9-15.
[2] KIYOHARA T, BOLLA R. Pathogenic variability among populations of the pinewood nematode, Bursaphelenchus xylophilus[J]. Forest Science, 1990, 36(4): 1061-76.
[3] 张星耀,骆有庆.中国森林重大生物灾害[M].北京:中国林业出版社, 2004.
[4] WEBSTER J M, ANDERSON R V, BAILLIE D L, et al. DNA probes for differentiating isolates of the pinewood nematode species complex[J]. Revue de Nématologie, 1990, 13(3): 255-263.
[5] 汤坚, 叶建仁, 陈凤毛,等. 松树体内寄生线虫种类及其致病性[J]. 南京林业大学学报(自然科学版), 2008, 32(3): 112-116. DOI: 10.3969/j.issn.1000-2006.2008.03.027.
TANG J, YE J R, CHEN F M, SU S R, et al. Species of parasitic nematode in pine host and pathogenicity[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2008, 32(3): 112-116.
[6] 张治宇,张克云,林茂松,等. 不同松材线虫群体对黑松的致病性测定[J]. 南京农业大学学报, 2002, 25(2): 43-46.
ZHANG Z Y, ZHANG K Y, LIN M S, et al. Pathogenicity determination of Bursaphelenchus xylophilus isolates to Pine thunbergii[J]. Journal of Nanjing Agricultural University,2002, 25(2): 43-46.
[7] 张治宇,林茂松,余本渊. 拟松材线虫对黑松苗的致病性[J].南京农业大学学报,2004,27(1): 46-50.
ZHANG Z Y, LIN M S, YU B Y. Pathogenicity of Bursaphelenchus mucronatus on the seedlings of black pine[J]. Journal of Nanjing Agricultural University,2004, 27(1): 46-50.
[8] 季宏铁.松材线虫(Bursaphelenchus xylophilus)与拟松材线虫(B. mucronatus)生物学杂交研究[D].南京:南京农业大学, 2004.
JI H T. Research of biological hybrid of Bursaphelenchus xylophilus and B. mucronatus[D]. Nanjing: Nanjing Agricultural University, 2004.
[9] 冯仕明,喻盛甫.拟松材线虫病危害不可忽视[J].云南林业, 2010(6): 29.
FENG S M, YU S F. Harm of Bursaphelenchus mucronatus is Assignable[J]. Yunnan Forest, 2010(6): 29.
[10] 陈凤毛,史延梅,王姝颖, 等. 不同株系拟松材线虫对黄山松和黑松苗木的致病性[J]. 林业科学, 2010, 46(12): 86-90. DOI:10.11707/j.1001-7488.20101214.
CHEN F M, SHI Y M, WANG S Y, et al. Pathogenicity of different isolates of Bursaphelenchus mucronatus to Pinus taiwanensis and P. thunbergii seedlings[J]. Scientia Silvae Sinicae, 2010, 46(12), 86-90.
[11] ZHOU L, CHEN F, PAN H, et al. Identifying virulence-associated genes using transcriptomic and proteomic association analyses of the plant parasitic nematode Bursaphelenchus mucronatus[J]. International Journal of Molecular Sciences, 2016, 17(9):1492. DOI: 10.3390/ijms17091492。
[12] KANG J S, LEE H, MOON I S, et al. Construction and characterization of subtractive stage-specific expressed sequence tag(EST)libraries of the pinewood nematode Bursaphelenchus xylophilus [J]. Genomics, 2009, 94(1): 70-77. DOI: 10.1016/j.ygeno.2009.03.001.
[13] 刘玉涛.松材线虫携带的细菌对其繁殖发育的影响[D].南京:南京林业大学,2003.
LIU Y T. The Effect of the bacteria carried by pine wood nematode Bursaphelenehus xylophilus on its reproduction and development[D]. Nanjing: Nanjing Forestry University, 2003.
[14] SINGH A, ROSENTHAL P J. Comparison of efficacies of cysteine protease inhibitors against five strains of Plasmodium falciparum[J]. Antimicrobial Agents and Chemotherapy, 2001, 45(3): 949-951. DOI: 10.1128/AAC.45.3.949-951.2001.
[15] WASILEWSKI M M, LIM K, PHILLIPS J, et al. Cysteine protease inhibitors block schistosome hemoglobin degradation in vitro and decrease worm burden and egg production in vivo[J]. Molecular and Biochemical Parasitology, 1996, 81(2): 179-189. DOI: 10.1016/0166-6851(96)02703-X.
[16] BERMAN J. Human leishmaniasis: clinical, diagnostic, and chemotherapeutic developments in the last 10 years[J]. Clinical Infectious Diseases, 1997, 24(4): 684-703.
[17] CARMONA C, DOWD A J, SMITH A M, et al. Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles[J]. Molecular and Biochemical Parasitology, 1993, 62(1): 9-17. DOI: 10.1016/0166-6851(93)90172-T.
[18] GARCIA M P, NBREGA O T, TEIXEIR A R, et al. Characterisation of a Trypanosoma cruzi acidic 30 kDa cysteine protease[J]. Molecular and biochemical parasitology, 1998, 91(2): 263-272. DOI:10.1016/S0166-6851(97)00205-3.
[19] SOLANO S, CORTES I M, COPITIN N I, et al. Lymphocyte apoptosis in the inflammatory reaction around Taenia solium metacestodes in porcine cysticercosis[J]. Veterinary Parasitology, 2006, 140(1): 171-176. DOI: 10.1016/j.vetpar.2006.03.006.
[20] SHINGLES J, LILLEY C, ATKINSON H, et al. Meloidogyne incognita: molecular and biochemical characterisation of a cathepsin L cysteine proteinase and the effect on parasitism following RNAi[J]. Experimental Parasitology, 2007, 115(2): 114-120. DOI: 10.1016/j.exppara.2006.07.008.
[21] NEVEU C, ABAD P, CASTAGNONE-S P. Molecular cloning and characterization of an intestinal cathepsin L protease from the plant-parasitic nematode Meloidogyne incognita [J]. Physiological and Molecular Plant Pathology, 2003, 63(3): 159-165. DOI: 10.1016/j.pmpp.2003.10.005.

相关文章 15

[1]	韦鹏飞, 理永霞, 冯宇倩, 刘振凯, 张星耀. 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.
[2]	江奕, 陈凤毛. 喜旱莲子草与乌蔹莓等植物提取物对松材线虫的毒杀活性研究[J]. 南京林业大学学报（自然科学版）, 2022, 46(4): 45-52.
[3]	吴佳雯, 尹艳楠, 谈家金, 郝德君. 蜡样芽孢杆菌NJSZ-13菌株诱导马尾松抗松材线虫病研究[J]. 南京林业大学学报（自然科学版）, 2022, 46(4): 53-58.
[4]	张瑞芝, 姜生伟, 吴昊, 陈俏丽, 李丹蕾, 王峰. 松材线虫Bx-HSF-1转录激活活性研究[J]. 南京林业大学学报（自然科学版）, 2022, 46(4): 8-14.
[5]	杨意, 刘波, 叶建仁, 苏禄晖. 水杉赤枯病综合营林生态控制技术研究[J]. 南京林业大学学报（自然科学版）, 2021, 45(6): 90-98.
[6]	高景斌, 徐六一, 叶建仁. 马尾松松材线虫病抗性无性系的筛选和遗传多样性分析[J]. 南京林业大学学报（自然科学版）, 2021, 45(5): 109-118.
[7]	何倩倩, 彭麒, 宗绡卓, 朱天辉, 李姝江. 蛋白毒素AP-Toxin诱导撑绿杂交竹抗梢枯病及其分泌物响应[J]. 南京林业大学学报（自然科学版）, 2020, 44(5): 199-208.
[8]	苏胜荣, 张巧巧, 叶建仁. 黄山地区垂柳叶锈病病原及发生规律研究[J]. 南京林业大学学报（自然科学版）, 2020, 44(4): 137-142.
[9]	尚征, 周立峰, 冯玥瑶, 刘费樱, 陈凤毛. 拟松材线虫个体发育研究[J]. 南京林业大学学报（自然科学版）, 2019, 43(6): 11-17.
[10]	丁森, 王焱, 陆蓝翔, 张岳峰, 江明明, 魏奇, 叶建仁. 一株促生抗病的樱花内生细菌的分离、筛选和鉴定[J]. 南京林业大学学报（自然科学版）, 2019, 43(5): 81-88.
[11]	戴美丽,方乐成,尹佟明,李小平. 基于SLAF-seq技术的抗杨树叶锈病SNP位点开发[J]. 南京林业大学学报（自然科学版）, 2019, 43(02): 73-78.
[12]	陈飞飞,黄金思,王翕韫,刘婉慧,叶建仁. 吡咯伯克霍尔德氏菌JK-SH007多聚半乳糖醛酸酶基因的克隆及表达分析[J]. 南京林业大学学报（自然科学版）, 2018, 42(04): 127-133.
[13]	李丹蕾,王峰,陈俏丽,张瑞芝,王佳楠. 美洲黑杨×毛果杨NDR1基因表达对E4锈菌侵染的响应[J]. 南京林业大学学报（自然科学版）, 2018, 42(01): 21-26.
[14]	朱涵明月,潘远智. 蓝花楹内生拮抗细菌筛选及对茎腐病的防治研究[J]. 南京林业大学学报（自然科学版）, 2017, 41(06): 61-67.
[15]	胡忠亮,郑催云,田兴一,李警保,韩正敏,樊奔. 解淀粉芽孢杆菌HZM9菌株发酵液的抑菌谱及稳定性测定[J]. 南京林业大学学报（自然科学版）, 2017, 41(03): 65-70.

拟松材线虫组织蛋白酶基因的RNA干扰及功能研究

Study on function of cathepsin bmcath1 in Bursaphelenchus mucronatus using method of RNA interference

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

作者

读者

审稿