[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):065-70.[doi:10.3969/j.issn.1000-2006.201612052]
 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):065-70.[doi:10.3969/j.issn.1000-2006.201612052]
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拟松材线虫组织蛋白酶基因的RNA干扰及功能研究
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
42
期数:
2018年05期
页码:
065-70
栏目:
研究论文
出版日期:
2018-09-15

文章信息/Info

Title:
Study on function of cathepsin bmcath1 in Bursaphelenchus mucronatus using method of RNA interference
文章编号:
1000-2006(2018)05-0065-06
作者:
潘妍毅12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
Author(s):
PAN Yanyi12 HUANG Lin1 WU Xiaoqin1*
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,
关键词:
拟松材线虫 组织蛋白酶基因 RNA干扰 繁殖力 致病力
Keywords:
Bursaphelenchus mucronatus cathepsin gene RNA interference reproduction ability pathogenicity
分类号:
S763.1
DOI:
10.3969/j.issn.1000-2006.201612052
文献标志码:
A
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.

参考文献/References:

[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.

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备注/Memo

备注/Memo:
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。
更新日期/Last Update: 2018-09-15