[1]李丹蕾,王 峰*,陈俏丽,等.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):021-26.[doi:10.3969/j.issn.1000-2006.201606028 ]
 LI Danlei,WANG Feng*,CHEN Qiaoli,et al.Impact of NDR1 gene on the incompatible Populus deltoidesP. trichocarpainfected with Melampsora larici-populina E4[J].Journal of Nanjing Forestry University(Natural Science Edition),2018,42(01):021-26.[doi:10.3969/j.issn.1000-2006.201606028 ]
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美洲黑杨×毛果杨NDR1基因表达对E4锈菌侵染的响应
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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01期
页码:
021-26
栏目:
研究论文
出版日期:
2018-01-31

文章信息/Info

Title:
Impact of NDR1 gene on the incompatible Populus deltoides×P. trichocarpa infected with Melampsora larici-populina E4
文章编号:
1000-2006(2018)01-0021-06
作者:
李丹蕾王 峰*陈俏丽张瑞芝王佳楠
东北林业大学林学院, 黑龙江 哈尔滨 150040
Author(s):
LI Danlei WANG Feng* CHEN Qiaoli ZHANG Ruizhi WANG Jianan
College of Forestry, Northeast Forestry University, Harbin 150040, China
关键词:
非小种专化抗病因子 亲和型互作关系 美洲黑杨×毛果杨 欧美杨 落叶松-杨栅锈菌
Keywords:
Keywords:NDR compatible interactions Populus deltoides×P. trichocarpa Populus×euramericana Melampsora larici-populina
分类号:
S763.15
DOI:
10.3969/j.issn.1000-2006.201606028
文献标志码:
A
摘要:
【目的】非小种专化抗病因子1(non-race-specific disease resistance 1,NDR1)可将病原入侵信号传入细胞膜内,继而激活含有CC结构域的CC-NBS-LRR抗性蛋白,介导植物抗性反应。探究杨树NDR1基因功能,克隆获得美洲黑杨×毛果杨(简称杂交杨)的PdtNDR1基因,并对PdtNDR1抗叶锈病基因表达进行研究。【方法】采用接种试验鉴定杂交杨和欧美杨与落叶松-杨栅锈菌E4强致病生理小种。采用PCR技术克隆杂交杨和欧美杨的NDR1基因(PdtNDR1和PndNDR1),并通过生物信息学分析其编码蛋白的结构与功能。采用荧光定量PCR技术分析锈菌接种后7个时间点PdtNDR1和PndNDR1的表达量变化。【结果】接种试验表明欧美杨是E4锈菌的感病品种,与E4锈菌构成亲和型互作关系。杂交杨是低感病品种,与E4锈菌构成非亲和型互作关系。生物信息学分析表明PdtNDR1和PndNDR1属于NHL家族。氨基酸序列分析表明PdtNDR1的N末端13—34位氨基酸和C末端嵌合在细胞膜上,中段氨基酸序列位于细胞外可以接触病原的侵染信号,N末端位于细胞内可将病原侵染信号传递到细胞内。荧光定量PCR技术检验锈菌接种后7个时间点PdtNDR1和PndNDR1对E4锈菌侵染的应激反应差异显著。PdtNDR1基因表达量在E4锈菌接种后持续增强,PndNDR1基因表达量未显著增加。【结论】PdtNDR1基因表达与杂交杨对E4锈菌的感病性正相关,决定杂交杨与锈菌的非亲和关系。
Abstract:
【Objective】The plant host resistance protein CC-NBS-LRR harboring CC domain could be activated to mediate plant resistance reaction after non-race-specific disease resistance 1(NDR1)and introduces pathogenic invasion signals into the cell membrane. To explore the function of NDR1, PdtNDR1 from Populus deltoids × P. trichocarpa was cloned and its expression on rust resistance was studied. 【Method】Compatible interactions of P. deltoides × P. trichocarpa and against Melampsora larici-populina E4 were observed by inoculation study. NDR1 genes, PdtNDR1 and PndNDR1 from P.deltoides × P.trichocarpa and P.×earamericama, respectively, were cloned by PCR. Structures and functions of PdtNDR1 and PndNDR1 encoded proteins were studied through bioinformatics analysis. The expressions of PdtNDR1 and PndNDR1 for 7 time points after being inoculated with E4 were analyzed by qPCR. 【Result】The inoculation study showed that P.×earame ricama was susceptible to E4, which was compatible to E4. P. deltoides × P. trichocarpa was a low susceptible species, which was incompatible to E4. The bioinformatics analysis indicated that PdtNDR1 and PndNDR1 belonged to NHL(NDR1/HIN1-like)family. Amino acid sequence analysis illustrated that 13-34 amino acids of the N-terminal and the C-terminal of PdtNDR1 located at the plasma membrane, the middle amino acids of PdtNDR1 located outside of the plasma membrane which could contact pathogenic invasion signals and the N-terminal of Pdt-NDR1 located inside of the cell enabling entrances of pathogenic invasion signals. qPCR results showed that PdtNDR1 and PndNDR1 responded to rust infection significantly differently. PdtNDR1 was enhanced continually after the rust infection, while the expression of PndNDR1 was not enhanced obviously. 【Conclusion】The expression of PdtNDR1 correlated with the susceptibility of P. deltoids × P.trichocarpa positively and determined its in compatible interaction against rust.

参考文献/References:

[1] DANGL J L, JONES J D G. Plant pathogens and integrated defence responses to infection [J]. Nature, 2001, 411:826-833. DOI: 10.1038/35081161.
[2] 牛春阳, 王峰, 李丹蕾, 等. C14族R2R3-MYB基因调控杨树抗锈菌过敏性反应 [J]. 北京林业大学学报, 2016, 38(7): 16-23. DOI: 10. 13332 /j. 1000—1522. 20150499. NIU C Y, WANG F, LI D L, et al. Impact of clade 14 R2R3-MYB genes on hypersensitive response of the poplar infected with Melampsora larici-populina[J]. Journal of Beijing Forestry University, 2016, 38(7): 16-23.
[3] CENTURY K S, HOLUB E B, STASKAWICZ B J. NDR1, a locus of Arabidopsis thaliana that is required for disease resistance to both a bacterial and a fungal pathogen [J]. Proc Nati Acad Sci USA, 1995, 92(14): 6597-6601. DOI: 10.1073/pnas.92.14.6597.
[4] 龚前园, 张超, 李为民, 等. 拟南芥NDR1基因介导的广谱抗病性研究进展 [J]. 生物技术通报, 2014(6):29-33. DOI:10.13560/j.cnki.biotech.bull.1985.2014.06.005. GONG Q Y, ZHANG C, LI W M, et al. Advances of NDR1 gene determined broad-spectrum disease resistance in Arabidopsis[J]. Biotechnology Bulletin, 2014(6):29-33.
[5] CHISHOLM S T, COAKER G, DAY B, et al. Host-microbe interactions: shaping the evolution of the plant immune response [J]. Cell, 2006, 124(4): 803-814. DOI: 10.1016/j.cell.2006.02.008.
[6] CACAS J L, PETITOT A S, BERNIER L, et al. Identification and characterization of the non-race specific disease resistance 1(NDR1)orthologous protein in coffee[J]. BMC Plant Biology, 2011, 11(1):1-17. DOI: 10.1186/1471-2229-11-144.
[7] MCNEECE B T, PANT S R, SHARMA K, et al. A Glycine max homolog of non-Race Specific Disease Resistance 1(NDR1)alters defense gene expression while functioning during a resistance response to different root pathogens in different genetic backgrounds[J]. Plant Physiology & Biochemistry, 2017, 114(5):60-71. DOI: 10.1016/j.plaphy.2017.02.022.
[8] 田呈明, 李振岐, 康振生. 青杨叶锈病(Melampsora larici-populina Kleb.)研究进展 [J]. 西北林学院学报, 1999, 14(2): 81-88. TIAN C M, LI Z Q, KANG Z S. Advances in researches of cathay poplar leaf rust(Melampsora larici-populina Kleb.)[J]. Journal of Northwest Forestry University, 1999, 14(2): 81-88.
[9] 曹支敏, 杜林, 王秦虎, 等. 中国落叶松-杨栅锈菌遗传多样性研究 [J]. 菌物学报, 2012, 31(4): 510-522. DOI:10.13346/j.mycosystema.2012.04.015. CAO Z M, DU L, WANG Q H, et al. Genetic diversity of poplar rust fungus Melampsora larici-populina in China [J]. Mycosystema, 2012, 31(4): 510-522.
[10] 陈俏丽, 李丹蕾, 王峰, 等. 欧美杂交杨Pnd-LRR3基因克隆及其抗锈菌侵染表达[J]. 东北林业大学学报, 2015, 43(3): 95-98. DOI:10.13759/j.cnki.dlxb.20150120.011. CHEN Q L,LI D L,WANG F,et al. Genetic cloning and gene expression of Pnd-LRR3 resistance gene from Populus nigra × P. deltoids [J]. Journal of Northeast Forestry University, 2015, 43(3): 95-98.
[11] LI D L,WANG F,WANG C,et al. MicroRNA-mediated susceptible poplar gene expression regulation associated with the infection of virulent Melampsora larici-populina [J]. Bmc Genomics, 2016, 17(1):1-16. DOI: 10.1186/s12864-015-2286-6.
[12] LI Q, LIN Y C, SUN Y H, et al. Splice variant of the SND1 transcription factor is a dominant negative of SND1 members and their regulation in Populus trichocarpa [J]. Proceedings of the National Academy of Sciences, 2012, 109(36): 14699-14704. DOI: 10.1073/pnas.1212977109.
[13] DUAN Y, JIANG Y, YE S, et al. PtrWRKY73, a salicylic acid-inducible poplar WRKY transcription factor, is involved in disease resistance in Arabidopsis thaliana [J]. Plant Cell Reports, 2015, 34(5): 831-841. DOI: 10.1007/s00299-015-1745-5.
[14] 李丹蕾, 张瑞芝, 王峰, 等. MicroRNA转录后调控欧美杨R2R3-MYBs抗锈菌表达 [J]. 林业科学研究, 2017, 30(2):254-259. DOI: 10.13275/j.cnki.lykxyj.2017.02.010. LI D L,ZHANG R Z,WAN F,et al. MicroRNA-regulated compatible poplar R2R3-MYB gene expression following the infection of virulent Melampsora larici-populina [J]. Forest Research, 2017, 30(2):254-259.
[15] 王峰, 程诗评, 牛春阳, 等. 杂交杨转录因子Pdt-ERF3抗锈病基因表达 [J]. 森林工程, 2016, 32(6):10-14. DOI:10.16270/j.cnki.slgc.2016.06.002. WANG F,CHENG S P,NIU C Y,et al. Pdt-ERF3 Expression of the hybrid poplar infected with Melampsora larici-populina[J]. Forest Engineering, 2016, 32(6):10-14.
[16] PEI M H, BAYON C, RUIZ C, et al. Population structure of poplar rust Melampsora larici-populina, in the UK inferred from AFLP[J]. Plant Pathology, 2007, 56(3):472-479. DOI: 10.1111/j.1365-3059.2007.01570.x.
[17] FREY P, PLNON J. Variability in pathogenicity of Melampsora allii-populina expressed on poplar cultivars [J]. Forest Pathology, 2010, 27(6):397-407. DOI: /10.1111/j.1439-0329.1997.tb01455.x.
[18] 王军. 植物的耐病性 [J]. 中国森林病虫, 1992(2):44-47. WANG J. Plant disease resistance [J]. Forest Pest and Disease, 1992(2):44-47.
[19] KNEPPER C, SAVORY E A, DAY B. Arabidopsis NDR1 is an integrin-like protein with a role in fluid loss and plasma membrane-cell wall adhesion [J]. Plant Physiol, 2011, 156: 286-300. DOI: 10.1104/pp.110.169656.
[20] LU H, ZHANG C, UTE A, et al. Overexpression of a citrus NDR1 ortholog increases disease resistance in Arabidopsis[J]. Frontiers in Plant Science, 2013, 4(2):157. DOI: 10.3389/fpls.2013.00157.
[21] KENICHI T, MASANAO S, JANE G, et al. Interplay between MAMP-triggered and SA-mediated defense responses [J]. Plant Journal for Cell & Molecular Biology, 2008, 53(5):763-775. DOI: 10.4161/psb.3.6.5702.
[22] DURRANT W E, DONG X. Systemic acquired resistance [J]. Annual Review of Phytopathology, 2004, 42(1): 185-209. DOI: 10.1146/annurev.phyto.42.040803.140421.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(31500524); 中央高校基本科研业务费专项资金项目(2572016CA04); 大学生创新性实验计划项目(201710225026) 第一作者:李丹蕾(danleili@nefu.edu.cn)。*通信作者:王峰(kingsummit@sina),副教授,博士。
更新日期/Last Update: 2018-03-30