欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2020, Vol. 46 ›› Issue (11): 1801-1809.doi: 10.3724/SP.J.1006.2020.04049

• 研究简报 • 上一篇    下一篇

低浓度呕吐毒素作为激发子对马铃薯抗干腐病的诱导及其作用机制

赵潇璨1(), 徐永清1, 贺付蒙1, 孙美丽1, 袁强1, 王雪1, 孔德兴1, 刘丹1, 冯艳忠2, 陈赫书2, 田明2, 刘娣2, 李凤兰1,*()   

  1. 1 东北农业大学生命科学学院, 黑龙江哈尔滨 150030
    2 黑龙江省农业科学院, 黑龙江哈尔滨 150086
  • 收稿日期:2020-02-28 接受日期:2020-07-02 出版日期:2020-11-12 网络出版日期:2020-07-17
  • 通讯作者: 李凤兰
  • 作者简介:E-mail:ZXC1656068953@163.com
  • 基金资助:
    本研究由国家自然科学基金青年项目(31201470);国家自然科学基金项目(J1210069);哈尔滨市应用科技研究与开发项目(2015RAQXJ021)

Low concentration of vomitoxin as elicitor induced resistance of dry rot disease of potato and its mechanism

ZHAO Xiao-Can1(), XU Yong-Qing1, HE Fu-Meng1, SUN Mei-Li1, YUAN Qiang1, WANG Xue1, KONG De-Xing1, LIU Dan1, FENG Yan-Zhong2, CHEN He-Shu2, TIAN Ming2, LIU Di2, LI Feng-Lan1,*()   

  1. 1 College of Life Science, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
    2 Heilongjiang Academy of Agricultural Sciences, Harbin 150086, Heilongjiang, China
  • Received:2020-02-28 Accepted:2020-07-02 Published:2020-11-12 Published online:2020-07-17
  • Contact: Feng-Lan LI
  • Supported by:
    This study was supported by the Youth Program of National Natural Science Foundation of China(31201470);the National Natural Science Foundation of China(J1210069);the Harbin Applied Science and Technology Research and Development Project(2015RAQXJ021)

RichHTML

11

PDF

372

摘要：

干腐病是马铃薯窖储过程中镰刀菌侵染引发的真菌病害, 严重影响了马铃薯的商品价值。呕吐毒素(vomitoxin, DON)又称脱氧雪腐镰刀菌烯醇, 是马铃薯干腐病致病镰刀菌在侵染薯块过程中产生的次生代谢产物。生物防治是病害防治的有效方法, 其中采用生物因子作为激发子诱导植物系统抗性的方法成为热门。本研究采用低浓度DON作为激发子对马铃薯块茎进行处理, 确定其在马铃薯抗干腐病中的作用及诱导马铃薯系统获得抗性(systemic acquired resistance, SAR)作用机制, 为马铃薯干腐病的生物防治提供理论依据。DON处理对马铃薯干腐病的扩展具有一定的影响, 且存在浓度效应, 其中5 ng mL-1DON处理马铃薯4 h能有效降低接种接骨木镰刀菌马铃薯块茎的干腐病病斑直径扩展; 低浓度DON处理提高了块茎组织的SOD、POD、几丁质酶、β-1,3-葡聚糖酶活性, 减少细胞膜过氧化产物MDA的积累; 薯块内的苯丙烷代谢关键酶PAL和4CL的活性升高, 促进了代谢产物总酚、类黄酮、木质素和花青素的积累。同时DON作为激发子可诱导马铃薯块茎中内源信号分子SA、JA和ET的含量增加, 植物系统抗性的调控基因NPR1的表达量上调。

关键词: 马铃薯, 干腐病, 镰刀菌, 呕吐毒素, 激发子, 诱导抗性

Abstract:

Dry rot disease is a fungal disease caused by Fusarium sp. infection in the process of potato cellar storage, which seriously affects the commercial value. Vomitoxin (DON), also known as deoxynivalenol, is a secondary metabolite produced by fusarium sp. in the process of infecting potato lump. Biological control is an effective method of disease control. Using biological factors as elicitors to induce plant system produce disease resistance has become a hot research topic. In this study, in order to provide theoretical basis for biological control of dry rot disease, potato tubers were treated with low concentration of DON as elicitor to determine its role in the resistance to dry rot disease and the mechanism of inducing systemic acquired resistance (SAR) of potato. DON treatment had a certain effect on the development of dry rot disease, and affected by the concentration. The treatment of potato tubers with 5 ng mL-1 DON for four hours could effectively reduce the diameter expansion of dry rot disease lesions induced by Fusarium sambucinum. Low concentration of DON increased the activities of SOD, POD, chitinase and β-1, 3-glucanase in tuber tissue, and decreased the accumulation of MDA. DON treatment increased the activities of PAL and 4CL, the key enzymes of phenylpropane metabolism, and promoted the accumulation of total phenols, flavonoids, lignin and anthocyanins. Meanwhile, the content of endogenous signal molecules SA, JA, and ET in potato tubers could be increased by using DON as elicitor, and the expression of NPR1 (regulatory gene of plant systemic resistance) was increased.

Key words: potato, dry rot disease, Fusarium sp., vomitoxin, elicitor, induced resistance

表1

Real time-PCR引物序列"

目的基因
Objective gene		 引物名称
Primer name		 引物序列
Primer sequence (5°-3°)
NPR1 NPR1-F CAGTGTGCTCGCCTATTTGTAT
NPR1-R GGTGTCGCTGAAACTTGTCG
Actin Actin-F GCTTCCCGATGGTCAAGTCA
Actin-R GGATTCCAGCTGCTTCCATTC

图1

不同浓度DON处理对马铃薯块茎接种接骨木镰刀菌病斑直径的影响 DON: 呕吐毒素。柱形图上不同小写字母表示差异显著(P < 0.05)。"

图2

DON处理6 h时接菌培养10 d后薯块内部形态变化 a~l分别代表无菌水、1 ng mL-1 DON、5 ng mL-1 DON、10 ng mL-1 DON、15 ng mL-1 DON、20 ng mL-1 DON、25 ng mL-1 DON、30 ng mL-1 DON、35 ng mL-1 DON、40 ng mL-1 DON、45 ng mL-1 DON、50 ng mL-1 DON处理。DON: 呕吐毒素。"

图3

DON处理不同时间对马铃薯块茎接种接骨木镰刀菌病斑大小的影响 DON: 呕吐毒素。柱形图上不同小写字母表示差异显著(P < 0.05)。"

图4

无菌水与DON处理后接菌培养10 d后薯块内部形态变化 DON: 呕吐毒素。"

图5

DON处理马铃薯块茎抗氧化性指标的变化 DON: 呕吐毒素; SOD: 超氧化物歧化酶; POD: 过氧化物酶; MDA: 丙二醛。*表示在0.05水平上显著; **表示在0.01水平上显著。"

图6

DON处理马铃薯块茎细胞壁防御酶的变化 DON: 呕吐毒素。*表示在0.05水平上显著; **表示在0.01水平上显著。"

图7

DON处理马铃薯块茎苯丙烷代谢关键酶活性及产物含量的变化 DON: 呕吐毒素。*表示在0.05水平上显著; **表示在0.01水平上显著。"

图8

DON处理马铃薯块茎内源激素含量的变化 DON: 呕吐毒素; SA: 水杨酸; JA: 茉莉酸; ET: 乙烯。*表示在0.05水平上显著; **表示在0.01水平显著。"

图9

DON处理马铃薯NPR1基因相对表达量变化 DON: 呕吐毒素。柱形图上不同小写字母表示差异显著(P < 0.05)。"

[1] 罗其友, 刘洋, 高明杰, 易晓峰. 中国马铃薯产业现状与前景. 农业展望, 2015,11(3):35-40.
Luo Q Y, Liu Y, Gao M J, Yi X F. Status quo and prospect of China’s potato industry. Agric Outlook, 2015,11(3):35-40 (in Chinese with English abstract).
[2] 陈彦云, 曹君迈, 陈晓军, 任晓月, 刘喜平, 丁红瑾. 我国马铃薯贮藏加工产业现状及其发展建议. 保鲜与加工, 2011,11(6):47-49.
Chen Y Y, Cao J M, Chen X J, Ren X Y, Liu X P, Ding H J. Present situation analysis and development suggestions on potato storage and processing industry of China. Storage Proc, 2011,11(6):47-49 (in Chinese with English abstract).
[3] 刁小琴, 关海宁, 魏雅冬, 马雪, 李杨, 郭丽, 马松艳. 窖藏马铃薯干腐病病原菌的分离及绿色防治技术研究. 食品研究与开发, 2015,36(1):122-125.
Diao X Q, Guan H N, Wei Y D, Ma X, Li Y, Guo L, Ma S Y. Study on isolation and green control technique of pathogens causing potato dry rot disease during storage. Food Res Dev, 2015,36(1):122-125 (in Chinese with English abstract).
[4] 敏凡祥, 王晓丹, 胡林双, 魏琪, 董学志, 李凤兰, 李鹤春, 刘伟婷, 郭梅. 黑龙江省马铃薯干腐病菌种类鉴定及致病性. 植物保护, 2010,36(4):112-115.
Min F X, Wang X D, Hu L S, Wei Q, Dong X Z, Li F L, Li H C, Liu W T, Guo M. Identification of species and pathogenicity of the Fusarium on potato in Heilongjiang province. Plant Prot, 2010,36(4):112-115 (in Chinese with English abstract).
[5] 李梅, 田世龙, 程建新, 李守强, 葛霞, 张欣. 三种防腐剂对马铃薯薯块干腐病菌的毒力及抑制作用. 植物保护, 2013,39(6):188-192.
Li M, Tian S L, Cheng J X, Li S Q, Ge X, Zhang X. Virulence and inhibition activity of three kinds of preservatives against the pathogen of the potato tuber dry rot. Plant Prot, 2013,39(6):188-192(in Chinese with English abstract).
[6] 陈亚兰, 张健. 5种药剂对贮藏期马铃薯干腐病防效试验. 甘肃农业科技, 2016, (3):42-44.
Chen Y L, Zhang J. Experiments of 5 kinds of pesticides on potato dry rot during storage. Gansu Agric Sci Technol, 2016, (3):42-44 (in Chinese).
[7] 侯忠艳. 马铃薯干腐病的发生与防治. 现代农业科技, 2012, (10):173-179.
Hou Z Y. Occurrence and control of potato dry rot. Modern Agric Sci Technol, 2012, (10):173-179 (in Chinese).
[8] Sadfi N, Chérif M, Fliss I, Boudabbous A, Antoun H. Evaluation of bacterial isolates from salty soils and bacillus thuringiensis strains for the biocontrol of Fusarium dry rot of potato tubers. J Plant Pathol, 2001,83:101-117.
[9] Jawed S, Mehmood A, Hussain A, Jatoi Z A, Kubar A A, Nizamani M M, Kaleri A A. Efficacy of different fungicides and bio control agents against Fusarium oxysporum, causal agent of potato dry rot. Indian J Sci Technol, 2019,12:1-12.
[10] 侯玉玲. 抗病诱导剂在蔬菜病害防治中的应用. 吉林蔬菜, 2017, (1):45-46.
Hou Y L. Application of disease resistance inducer in vegetable disease control. Jilin Veget, 2017, (1):45-46 (in Chinese).
[11] 李群. 钙依赖蛋白激酶和腺苷酸环化酶参与拟南芥响应大丽轮枝菌毒素诱导信号转导过程的实验证据. 中国农业大学博士学位论文, 北京, 2005.
Li Q. Evidence for Involvment of Calcium-dependent Protein Kinase and Adenylyl Cyclase in Verticillium Toxin Induced Signal Transduction in Arabidopsis thaliana. PhD Dissertation of China Agricultural University, Beijing,China, 2005 (in Chinese with English abstract).
[12] 赵小明, 杜昱光. 寡糖激发子及其诱导植物抗病性机理研究进展. 中国农业科技导报, 2006,8(6):26-32.
Zhao X M, Du Y G. Progress of research on oligosaccharide elicitors and mechanism of plant induced resistance by oligosaccharides. Rev China Agric Sci Technol, 2006,8(6):26-32 (in Chinese with English abstract).
[13] Block A, Schmelz E A, O’Donnell P J, Jones J, Kiee H J. Systemic acquired tolerance to virulent bacterial pathogens in tomato. Plant Physiol, 2005,138:1481-1490.
doi: 10.1104/pp.105.059246 pmid: 15937273
[14] Dong X. SA, JA, ethylene and disease resistance in plants. Curr Opin Plant Biol, 1998,1:316-323.
pmid: 10066607
[15] 何苏琴, 金秀琳, 魏周全, 张廷义, 杜玺, 骆得功. 甘肃省定西地区马铃薯块茎干腐病病原真菌的分离鉴定. 云南农业大学学报, 2004,19:550-552.
He S Q, Jin X L, Wei Z Q, Zhang T Y, Du X, Luo D G. Isolation and identification of pathogens causing dry rot of potato tuber in Dingxi prefecture of Gansu province. J Yunnan Agric Univ, 2004,19:550-552 (in Chinese with English abstract).
[16] Bajestani B, Bastam V. Study on differentially lipase gene expression against different inducers of defense response in scab-resistant and susceptible wheat cultivars. J Rec Adv Agric, 2012,1:6-14.
[17] 潘哲超, 陈建斌, 范静华, 周惠萍, 邱世刚, 叶漪, 果志华. 稻瘟菌粗毒素对水稻防御性相关酶系的诱导. 云南农业大学学报, 2008,23(2):28-32.
Pan Z C, Chen J B, Fan J H, Zhou H P, Qiu S G, Ye Y, Guo Z H. Activities of defense-related enzymes induced by grude toxin from Magnaporthe grisea. J Yunnan Agric Univ, 2008,23(2):28-32 (in Chinese with English abstract).
[18] 马春红, 翟彩霞, 商闯, 贾银锁. 低浓度玉米小斑病菌T小种和C小种毒素培养滤液对玉米C103叶片过氧化物酶活性的诱导作用见: 彭友良, 王振中主编. 中国植物病理学会2008年学术年会论文集. 北京: 中国农业科学技术出版社, 2008. pp 480-484.
Ma C H, Zhai C X, Shang C, Jia Y S. The induced effects of filtrate of bipolaris maydis race C toxin and race T toxin cultivation on POD activities in maize leaves C103. In: Peng Y L, Wang Z Z, eds. Proceedings of the Annual Meeting of Chinese Society for Plant Pathology (2008). Beijing: China Agricultural Science and Technology Press, 2008. pp 480-484(in Chinese).
[19] 张志良. 植物生理学实验指导. 北京: 高等教育出版社, 2003. pp 123-124, 268-268, 274-276.
Zhang Z L. Plant Physiology Experiment Guide. Beijing: Higher Education Press, 2003. pp 123-124, 268-268, 274-276(in Chinese).
[20] 上海市植物生理学会. 现代植物生理学实验指南. 上海: 上海科学技术出版社, 1999. pp 129-130.
Shanghai Plant Physiology Association. Handbook of Plant Physiology Experiment. Shanghai: Shanghai Scientific and Technical Publishers, 1999. pp 129-130(in Chinese).
[21] 曹建康, 姜微波, 赵玉梅. 果蔬采后生理生化实验指导. 北京: 中国轻工业出版社, 2007. pp 44-45, 142-143.
Cao J K, Jiang W B, Zhao Y M. Guidance on Post-harvest Physiological and Biochemical Experiments of Fruits and Vegetables . Beijing: China Light Industry Press, 2007. pp 44-45, 142-143(in Chinese).
[22] 杨志敏, 毕阳, 李永才, 寇宗红, 包改红, 刘成琨, 王毅, 王蒂. 马铃薯干腐病菌侵染过程中切片组织细胞壁降解酶的变化. 中国农业科学, 2012,45:127-134.
Yang Z M, Bi Y, Li Y C, Kou Z H, Bao G H, Liu C K, Wang Y, Wang D. Changes of cell wall degrading enzymes in potato tuber tissue slices infected by Fusarium sulphureum. Sci Agric Sin, 2012,45:127-134 (in Chinese with English abstract).
[23] 刘玉良, 米福贵, 特木尔布和, 王普昶, 马小廷. 苜蓿叶片水杨酸含量与其蓟马抗性的关系. 西北植物学报, 2011,31:588-594.
Liu Y L, Mi F G, Temu’erbuhe , Wang P C, Ma X T. Relationship between alfalfa salicylic acid content and Its thrips resistance. Acta Bot Boreali-Occident Sin, 2011,31:588-594 (in Chinese with English abstract).
[24] 侯志强, 王庆国. 鲜切马铃薯实时荧光定量PCR分析中内参基因的选择. 安徽农业科学, 2013,41:5207-5209.
Hou Z Q, Wang Q G. Reference genes selection for real-time fluorescence quantitative PCR in fresh-cut potato. J Anhui Agric Sci, 2013,41:5207-5209 (in Chinese with English abstract).
[25] Lazarovits G, Higgins V J. Biological activity and specificity of a toxin produced by Cladosporium fulvum. Physiol Biochem, 1979,69:1056-1061.
[26] 王桥美, 范静华, 果志华, 陈建斌. 烟草黑胫病菌毒素对烟草防御性相关酶的诱导作用. 云南农业大学学报, 2011,26:20-25.
Wang Q M, Fan J H, Guo Z H, Chen J B. Induction effect of toxin extracts of Phytophthora nicotianae on tobacco defense- related enzyme activities. J Yunnan Agric Univ, 2011,26:20-25 (in Chinese with English abstract).
[27] Nishiuchi T, Masuda D, Nakashita H, Ichimura K, Shinozaki K, Yoshida S, Kimura M, Yamaguchi I, Yamaguchi K. Fusarium phytotoxin trichothecenes have an elicitor-like activity in Arabidopsis thaliana, but the activity differed significantly among their molecular species. Mol Plant-Microbe Interact, 2006,19:512-520.
pmid: 16673938
[28] Bhattacharjee S. Reactive oxygen species and oxidative burst: roles in stress, senescence and signal transduction in plants. Curr Sci, 2005,89:1113-1121.
[29] 史娟, 李建设. 枯萎病菌诱导的几丁质酶和β-1,3-葡聚糖酶与寄主抗病性的关系. 农业科学研究, 2006,27(3):24-26.
Shi J, Li J S. Relationship between chitinase and β-1,3-glacanase activity induced by Fusarium oxysporum on resistance of cucumber to Fusarium Wilt. J Agric Sci, 2006,27(3):24-26 (in Chinese with English abstract).
[30] 邓惠文, 毕阳, 葛永红, 任亚琳, 刘瑶瑶. 采后BTH处理及粉红单端孢(Trichothecium roseum)挑战接种对厚皮甜瓜果实苯丙烷代谢活性的诱导. 食品工业科技, 2013,34(1):323-326.
Deng H W, Bi Y, Ge Y H, Ren Y L, Liu Y Y. Induction of phenylpropanoid metabolic activity in muskmelon fruit by postharvest BTH treatment and challenge inoculation with Trichothecium roseum. Sci Technol Food Ind, 2013,34(1):323-326 (in Chinese with English abstract).
[31] 宋阳, 王丕武, 张学明, 曲静. 植物广谱抗病基因NPR1的研究进展. 农业与技术, 2013,33(6):5-7.
Song Y, Wang P W, Zhang X M, Qu J. Research progress on plant broad-spectrum disease resistance gene NPR1. Agric Technol, 2013,33(6):5-7 (in Chinese).
[32] Aver’yanov A A, Lapikova V P, Lebrun M H. Tenuazonic acid, toxin of rice blast fungus, induces disease resistance and reactive oxygen production in plants. Russ J Plant Physiol, 2007,54:749-754.
[33] Xue H, Bi Y, Prusky D, Raza H, Zhang R, Zhang S, Nan M, Zong Y Y, Cheng X Y. The mechanism of induced resistance against Fusarium dry rot in potato tubers by the T-2 toxin. Posth Biol Technol, 2019,153:69-78.
[34] 中华人民共和国卫生部. GB/T 2761-2017食品安全国家标准食品中真菌毒素限量. 北京: 中国标准出版社, 2017. pp 1-3.
The Minister of Health of the People’s Republic of China. GB/T 2761-2017 National Food Safety Standard Food Mycotoxins Limits. Beijing: Standards Press of China, 2017. pp 1-3(in Chinese).
[1] 王海波, 应静文, 何礼, 叶文宣, 涂卫, 蔡兴奎, 宋波涛, 柳俊. rDNA和端粒重复序列鉴定马铃薯和茄子体细胞杂种染色体丢失和融合[J]. 作物学报, 2022, 48(5): 1273-1278.
[2] 石艳艳, 马志花, 吴春花, 周永瑾, 李荣. 垄作沟覆地膜对旱地马铃薯光合特性及产量形成的影响[J]. 作物学报, 2022, 48(5): 1288-1297.
[3] 冯亚, 朱熙, 罗红玉, 李世贵, 张宁, 司怀军. 马铃薯StMAPK4响应低温胁迫的功能解析[J]. 作物学报, 2022, 48(4): 896-907.
[4] 张霞, 于卓, 金兴红, 于肖夏, 李景伟, 李佳奇. 马铃薯SSR引物的开发、特征分析及在彩色马铃薯材料中的扩增研究[J]. 作物学报, 2022, 48(4): 920-929.
[5] 谭雪莲, 郭天文, 胡新元, 张平良, 曾骏, 刘晓伟. 黄土高原旱作区马铃薯连作根际土壤微生物群落变化特征[J]. 作物学报, 2022, 48(3): 682-694.
[6] 余慧芳, 张卫娜, 康益晨, 范艳玲, 杨昕宇, 石铭福, 张茹艳, 张俊莲, 秦舒浩. 马铃薯CrRLK1Ls基因家族的鉴定及响应晚疫病菌信号的表达分析[J]. 作物学报, 2022, 48(1): 249-258.
[7] 荐红举, 尚丽娜, 金中辉, 丁艺, 李燕, 王季春, 胡柏耿, Vadim Khassanov, 吕典秋. 马铃薯PIF家族成员鉴定及其对高温胁迫的响应分析[J]. 作物学报, 2022, 48(1): 86-98.
[8] 许德蓉, 孙超, 毕真真, 秦天元, 王一好, 李成举, 范又方, 刘寅笃, 张俊莲, 白江平. 马铃薯StDRO1基因的多态性鉴定及其与根系性状的关联分析[J]. 作物学报, 2022, 48(1): 76-85.
[9] 唐锐敏, 贾小云, 朱文娇, 印敬明, 杨清. 马铃薯热激转录因子HsfA3基因的克隆及其耐热性功能分析[J]. 作物学报, 2021, 47(4): 672-683.
[10] 李鹏程, 毕真真, 孙超, 秦天元, 梁文君, 王一好, 许德蓉, 刘玉汇, 张俊莲, 白江平. DNA甲基化参与调控马铃薯响应干旱胁迫的关键基因挖掘[J]. 作物学报, 2021, 47(4): 599-612.
[11] 秦天元, 刘玉汇, 孙超, 毕真真, 李安一, 许德蓉, 王一好, 张俊莲, 白江平. 马铃薯StIgt基因家族的鉴定及其对干旱胁迫的响应分析[J]. 作物学报, 2021, 47(4): 780-786.
[12] 蒋伟, 潘哲超, 包丽仙, 周福仙, 李燕山, 隋启君, 李先平. 马铃薯资源晚疫病抗性的全基因组关联分析[J]. 作物学报, 2021, 47(2): 245-261.
[13] 柳燕兰, 郭贤仕, 张绪成, 马明生, 王宏康. 密度和施肥对旱地马铃薯干物质积累、产量和水肥利用的影响[J]. 作物学报, 2021, 47(2): 320-331.
[14] 牛娜, 刘震, 黄鹏翔, 朱金勇, 李志涛, 马文婧, 张俊莲, 白江平, 刘玉汇. 马铃薯GAUT基因家族的全基因组鉴定及表达分析[J]. 作物学报, 2021, 47(12): 2348-2361.
[15] 吴春花, 普雪可, 周永瑾, 勉有明, 苗芳芳, 李荣. 宁南旱区沟垄集雨结合覆盖对土壤水热肥与马铃薯产量的影响[J]. 作物学报, 2021, 47(11): 2208-2219.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2