[1]邓丽娜,吴小芹*.松材线虫毒力及发育转型与细胞自噬关系研究[J].南京林业大学学报(自然科学版),2016,40(04):171-176.[doi:10.3969/j.issn.1000-2006.2016.04.027]
 DENG Lina,WU Xiaoqin*.Research on the relationship of autophagy with the virulence and development of Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2016,40(04):171-176.[doi:10.3969/j.issn.1000-2006.2016.04.027]
点击复制

松材线虫毒力及发育转型与细胞自噬关系研究
分享到:

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
40
期数:
2016年04期
页码:
171-176
栏目:
研究简报
出版日期:
2016-08-30

文章信息/Info

Title:
Research on the relationship of autophagy with the virulence and development of Bursaphelenchus xylophilus
文章编号:
1000-2006(2016)04-0171-06
作者:
邓丽娜12吴小芹1*
1.南方现代林业协同创新中心,南京林业大学林学院,江苏省有害生物入侵预防与控制重点实验室, 江苏 南京 210037;
2.盐城工学院海洋与生物工程学院,江苏 盐城 224051
Author(s):
DENG Lina12 WU Xiaoqin1*
1. Co-Innovation Center for the Sustainable Forestry in Southern China, Jiangsu Key Laboratory for Pest Prevention and Management of Invasive Specie, College of Forestry, Nanjing Forestry University, Nanjing 210037, China;
2. School of Ocean and Biological Engineering, Yancheng Institute of Technology, Yancheng 224051,China
关键词:
松材线虫 细胞自噬 毒力 发育转型 透射电镜 实时定量PCR
Keywords:
Bursaphelenchus xylophilus autophagy virulence development transmission electron microscopy RT-qPCR
分类号:
S763; Q958.9
DOI:
10.3969/j.issn.1000-2006.2016.04.027
文献标志码:
A
摘要:
采用透射电镜法对松材线虫的细胞自噬现象进行鉴认,并通过实时定量PCR的方法对不同毒力及不同发育期松材线虫的自噬基因BxAtg1和BxAtg8表达量进行测定。结果表明:饥饿诱导下不同毒力松材线虫存在细胞自噬现象,不同毒力松材线虫饥饿诱导12、24、36 h时,自噬基因BxAtg1表达量递减且趋势一致,而自噬基因BxAtg8在饥饿24 h时的强毒松材线虫体内表达量最高,在饥饿36 h时弱毒松材线虫体内表达量显著升高; 与繁殖性松材线虫体相比,自噬基因BxAtg1和BxAtg8在扩散型3龄(J)和扩散型4龄幼虫(J)中的表达量均增加,推测自噬活性的调节机制可能与松材线虫的毒力相关,与松材线虫发育转型阶段的形态特征联系,暗示自噬可帮助松材线虫发育转型,从而提高松材线虫的适应性。
Abstract:
In order to identify the autophagy in Bursaphelenchus xylophilus, research on the relationship between autophagy and the different virulence and development of B. xylophilus was carried out in this paper. Transmission electron microscopy(TEM)method was used to identify autophagy in B. xylophilus, and quantitative reverse transcription PCR(qRT-PCR)was adopted to quantify the BxAtg1 and BxAtg8 at different virulence and at different stages of the development of B. xylophilus. The results from TEM observations showed that the process of autophagy existed in B. xylophilus after starvation-induced. The mRNA expression of BxAtg1 was decreased in different virulence nematodes which exposed to long-term starvation for 12, 24 and 36 h. The mRNA expression of BxAtg8 in strongly virulent nematodes was increased in 24 h, but the mRNA expression of BxAtg8 in weakly virulent nematodes was obviously increased in 36 h. And there was increment of both the mRNA expression of BxAtg1 and BxAtg8 in J and Jcompared to the propagtive form of B. xylophilus. Compareing to the morphological characteristics of the different development of Bursaphelenchus xylophilus. These suggest that the mechanism of autophagy activity regulating may be related to the virulence of nematodes and autophagy assisted the nematodes in entering dispersal phase, which improved their ability to adapt to the environment at the same time. This study provided fundamental information the autophagy of B. xylophilus.

参考文献/References:

[1] 叶建仁. 松材线虫病诊断与防治技术[M]. 北京:中国林业出版社, 2010.
[2] 杨宝君. 松材线虫病[M]. 北京: 中国林业出版社, 2003:1-25.
[3] Mamiya Y. Pathology of the pine wilt disease caused by Bursaphelenchus xylophilus[J]. Annu Rev Phytopathol, 1983, 21:201-220. Doi:10.1146/annurev.py.21.090183.001221.
[4] Wingfield M J, Blanchette R A, Nicholls T H, et al. The pine wood nematode: a comparison of the situation in the United States and Japan[J]. Canadian Journal of Forest Research, 1982, 12(1):71-75. Doi:10.1139/x82-010.
[5] 杨振德, 赵博光. 松材线虫行为学研究进展[J]. 南京林业大学学报(自然科学版), 2003, 27(1): 87-92. Doi:10.3969/j.issn.1000-2006.2003.01.020. Yang Z D, Zhao B G. Review on behavior studies of the pine wood nematode[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2003, 27(1): 87-92.
[6] 王玲萍. 松墨天牛生物学特性的研究[J]. 福建林业科技, 2004, 31(3): 23-26. Doi:10.3969/j.issn.1002-7351.2004.03.006. Wang L P. Study on the biological characteristic of Monochamus alternatus Hope[J]. Journal of Fujian Forestry Science And Technology, 2004, 31(3): 23-26.
[7] Reggiori F, Klionsky D J. Autophagy in the eukaryotic cell[J]. Eukaryotic Cell, 2002, 1(1): 11–21.
[8] 张磊. 稻瘟病菌相关致病基因MgATG3、MgATG4和MgATG7的克隆和功能分析[D]. 杭州: 浙江大学, 2007. Zhang L. The molecular characterization and functional analysis of pathogenic related gene MgATG3、MgATG4 and MgATG7 from Magnaporthe grisea[D]. Hangzhou: Zhejiang University, 2007.
[9] Asakura M, Ninomiya S, Sugimoto M, et al. Atg26-mediated pexophagy is required for host invasion by the plant pathogenic fungus colletotrichum orbiculare[J]. The Plant Cell Online, 2009, 21(4):1291-1304. Doi:10.1105/tpc.108.060996.
[10] 李超萍, 林春花, 翟李刚, 等. 橡胶树胶孢炭疽病菌致病相关基因CgATG8的功能分析[J]. 热带作物学报, 2013,34(11):2172-2178. Doi:10.3969/j.issn.1000-2561.2013.11.017. Li C P, Lin C H, Zhai L G, et al. Functional analysis of pathogenic related gene CgATG8 from Colletotrichum gloeosporioides[J]. Chinese Journal of Tropical Crops, 2013, 34((11)):2172-2178.
[11] Veneault-Fourrey C, Barooah M, Egan M, et al. Autophagic fungal cell death is necessary for infection by the rice blast fungus[J]. Science, 2006, 312:580-583. Doi:10.1126/science.1124550.
[12] 朱克恭. 松材线虫病研究综述[J]. 世界林业研究, 1995(4): 28-33. Zhu K G. A review of studies on pine wilt disease caused by Bursaphelenchus xylophilus[J]. World Forestry Research, 1995(4): 28-33.
[13] 张宾,胡春祥,石进,等. 线虫转型发育和寄主识别的化学通讯研究进展[J]. 生态学报, 2013, 33(7): 2003-2012. Doi:10.5846/stxb201112241967. Zhang B,Hu C X,Shi J, et al. Research progress on chemical communication of development and host-finding of nematodes[J]. Acta Ecologica Sinica, 2013, 33(7): 2003-2012.
[14] Alberti A, Michelet X, Djeddi A, et al. The autophagosomal protein LGG-2 acts synergistically with LGG-1 in dauer formation and longevity in C. elegans[J]. Autophagy, 2010, 6(5):622-633. Doi:10.4161/auto.6.5.12252.
[15] Melendez A. Autophagy genes are essential for dauer development and life-span extension in C. elegans[J]. Science, 2003:1387-1391. 10.1126/science.1087782.
[16] 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. Doi:10.3321/j.issn:1000-2030.2002.02.010. 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.
[17] 朱丽华, 施学文, 胡玉丹,等. 不同毒力松材线虫杂交后代的繁殖力和致病力的比较[J]. 南京林业大学学报(自然科学版), 2015, 39(1):1-5. Doi:10.3969/j.issn.1000-2006.2015.01.001. Zhu L H, Shi X W, Hu Y D, et al. The fecundity and virulence of offspring crossbred from virulent and avirulent isolates of Bursaphelenchus xylophilus[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2015, 39(1):1-5.
[18] Deng L N, Wu X Q, Ye J R, et al. Identification of autophagy in the pine wood nematode Bursaphelenchus xylophilus and the molecular characterization and functional analysis of two novel autophagy-related genes, BxATG1 and BxATG8[J]. Ijms, 2016, 17(3):279. Doi:10.3390/ijms17030279.
[19] 刘雪莲. 中国松材线虫群体形态学及致病性变异的研究[D]. 南京: 南京林业大学, 2007. Liu X L. Morphology and pathogenic mutation of Bursaphelenchus xylophilus in china[D]. Nanjing: Nanjing Forestry University, 2007.
[20] Levine B, Kroemer G. Autophagy in the pathogenesis of disease[J]. Cell, 2008, 132(1):27-42. Doi:10.1016/j.cell.2007.12.018.
[21] Mizushima N. Autophagy: process and function[J]. Genes Dev, 2007, 21(22):2861-2873. Doi:10.1101/gad.1599207.
[22] 王威, 陈景红, 王新宁, 等. 葡萄籽原花青素诱导人肝癌HepG2细胞凋亡及自噬性死亡[J]. 暨南大学学报(自然科学与医学版), 2011, 32(2): 181-187. Doi:10.3969/j.issn.1000-9965.2011.02.014. Wang W, Chen J H, Wang X N,et al. Grape seed proanthocyanidins induce apoptosis and autophagic cell death in human hepatoma HepG2 cells[J]. Journal of Jinan University(Natural Science & Medicine Edition), 2011, 32(2): 181-187.
[23] 高玉仁, 张德福, 王梁,等. 桧木醇激活膀胱癌J82细胞自噬诱导细胞凋亡[J]. 中国癌症杂志, 2015(5): 365-370. Doi:10.3969/j.issn.1007-3969.2015.05.008. Gao Y R, Zhang D F, Wang L, et al. Hinokitiol induces bladder cancer J82 cells apoptosis via autophagy induction[J]. China Oncology, 2015(5): 365-370.
[24] Shpilka T, Weidberg H, Pietrokovski S, et al. Atg8: an autophagy-related ubiquitin-like protein family[J]. Genome Biol, 2011, 12(7):226. Doi:10.1186/gb-2011-12-7-226.
[25] Xie Z, Nair U, Klionsky D J. Atg8 controls phagophore expansion during autophagosome formation[J]. Mol Biol Cell, 2008, 19(8):3290-3298. Doi:10.1091/mbc.E07-12-1292.
[26] Aladzsity I, Tóth M L, Sigmond T, et al. Autophagy genes unc-51 and bec-1 are required for normal cell size in Caenorhabditis elegans[J]. Genetics, 2007, 177(1):655-660. Doi:10.1534/genetics.107.075762.

相似文献/References:

[1]朱丽梅,吴小芹*,蒋 萍.细菌XSJS3对松材线虫杀线活性的测定[J].南京林业大学学报(自然科学版),2008,32(01):083.[doi:10.3969/j.jssn.1000-2006.2008.01.019]
 ZHU Li-mei,WU Xiao-qin*,JIANG Ping.A study of the nematicidal activity of bacterium XSJS3 to Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2008,32(04):083.[doi:10.3969/j.jssn.1000-2006.2008.01.019]
[2]高景斌,徐六一,叶建仁*,等.运用MuPS标记识别马尾松抗松材线虫病个体[J].南京林业大学学报(自然科学版),2009,33(03):001.[doi:10.3969/j.jssn.1000-2006.2009.03.001]
 GAO Jing bin,XU Liu yi,YE Jian ren*,et al.Using MuPS marker technique to identify individual of Pinus massoniana resistance to pine wilt disease[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(04):001.[doi:10.3969/j.jssn.1000-2006.2009.03.001]
[3]刘劼,吴小芹*,应晨希,等.松材线虫和拟松材线虫的繁殖力及其超氧自由基差异[J].南京林业大学学报(自然科学版),2009,33(03):005.[doi:10.3969/j.jssn.1000-2006.2009.03.002]
 LIU Jie,WU Xiao qin*,YING Chen xi,et al.The difference of progenitive power and superoxide anion production in Bursaphelenchus xylophilus and B.mucronatus[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(04):005.[doi:10.3969/j.jssn.1000-2006.2009.03.002]
[4]黄麟,叶建仁*,刘雪莲.松材线虫病病原种群分化研究现状[J].南京林业大学学报(自然科学版),2009,33(04):135.[doi:10.3969/j.jssn.1000-2006.2009.04.030]
 HUANG Lin,YE Jian ren*,LIU Xue lian.Advance in population differentiation of Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(04):135.[doi:10.3969/j.jssn.1000-2006.2009.04.030]
[5]金钢,叶建仁*.不同致病力线虫侵染后黑松的水势和根系活力的变化[J].南京林业大学学报(自然科学版),2009,33(06):053.[doi:10.3969/j.jssn.1000-2006.2009.06.012]
 JIN Gang,YE Jian ren*.Study on the changes in the flow of water and the activity of roots system after Pinus thunbergii inoculated with Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(04):053.[doi:10.3969/j.jssn.1000-2006.2009.06.012]
[6]瞿红叶,谈家金*,叶建仁,等.不同真菌对松材线虫繁殖及致病力的影响[J].南京林业大学学报(自然科学版),2009,33(06):057.[doi:10.3969/j.jssn.1000-2006.2009.06.013]
 QU Hong ye,TAN Jia jin*,YE Jian ren,et al.Effects of different fungus on the reproduction and virulence of Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(04):057.[doi:10.3969/j.jssn.1000-2006.2009.06.013]
[7]赵博光.松材线虫携带的细菌在松萎蔫病中的作用[J].南京林业大学学报(自然科学版),2010,34(01):145.[doi:10.3969/j.jssn.1000-2006.2010.01.033]
 ZHAO Bo guang.The role of bacteria carried by Bursaphelenchus xylophilus in pine wilt disease[J].Journal of Nanjing Forestry University(Natural Science Edition),2010,34(04):145.[doi:10.3969/j.jssn.1000-2006.2010.01.033]
[8]Kazuyoshi FUTAI.对松萎蔫病的综合理解[J].南京林业大学学报(自然科学版),2010,34(01):148.[doi:10.3969/j.jssn.1000-2006.2010.01.034]
 Kazuyoshi FUTAI.A comprehensive understanding of pine wilt disease[J].Journal of Nanjing Forestry University(Natural Science Edition),2010,34(04):148.[doi:10.3969/j.jssn.1000-2006.2010.01.034]
[9]P.Vieira,M.Mota.2006—2009年葡萄牙松材线虫(Bursaphelenchus xylophilus)研究进展[J].南京林业大学学报(自然科学版),2010,34(01):150.[doi:10.3969/j.jssn.1000-2006.2010.01.035]
 P.Vieira,M.Mota.Progress in research on the pinewood nematode (Bursaphelenchus xylophilus),[J].Journal of Nanjing Forestry University(Natural Science Edition),2010,34(04):150.[doi:10.3969/j.jssn.1000-2006.2010.01.035]
[10]Ryoji Shinya,Yuko Takeuchi,Hironobu Morisaka,等.松材线虫的表被层蛋白[J].南京林业大学学报(自然科学版),2010,34(02):150.[doi:10.3969/j.jssn.1000-2006.2010.02.035]
 Ryoji Shinya,Yuko Takeuchi,Hironobu Morisaka,et al.Surface coat proteins of the pine wood nematode, Bursaphelenchus xylophilus[J].Journal of Nanjing Forestry University(Natural Science Edition),2010,34(04):150.[doi:10.3969/j.jssn.1000-2006.2010.02.035]
[11]吴 凡,吴小芹*.细胞自噬对松材线虫繁殖存活的影响及线虫与松树互作早期自噬相关基因的表达[J].南京林业大学学报(自然科学版),2018,42(05):059.[doi:10.3969/j.issn.1000-2006.201712025]
 WU Fan,WU Xiaoqin*.The role of autophagy of pine wood nematode Bursaphelenchus xylophilus in the growth and development and infecting pine trees at early time[J].Journal of Nanjing Forestry University(Natural Science Edition),2018,42(04):059.[doi:10.3969/j.issn.1000-2006.201712025]

备注/Memo

备注/Memo:
收稿日期:2015-10-15 修回日期:2016-03-17
基金项目:国家自然科学基金项目(31270683); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:邓丽娜(624174349@qq.com),博士。*通信作者:吴小芹(xqwu@njfu.com.cn),教授。
引文格式:邓丽娜,吴小芹. 松材线虫毒力及发育转型与细胞自噬关系研究[J]. 南京林业大学学报(自然科学版),2016,40(4):171-176.
更新日期/Last Update: 2016-08-30