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Agricultural Sciences in China 2007年第1期6卷 78-85页

作者： FAN Yong-shan gu shou-qin DONG Jin-gao DONG Bing-fang Mycotoxin Laboratory Agricultural University of Hebei Baoding 071001 P.R.China Department of Biological Science and Technology Tangshan Teachers College Tangshan 063000 PR.China

Systemic studies on the effects of mitogen-activated protein kinase （MAPK） signal transduction pathway on the growth and development of Setosphaeria turcica is helpful not only in understanding the molecular mechanism of pathogenhost interaction but also in the effective control of the diseases caused by S. turcica. U0126, the specific MEK inhibitor, is used to treat S. turcica before the observation of the conidial germination, appressorium production, and pathogenicity of the pathogen. There is no significant effect of U0126 on the colony morphology and mycelium growth of the pathogen. After treatment with U0126, the growth of mycelium and conidia are normal, but the conidial germination, appressorium production, and pathogenicity of S. turcica on susceptible corn leaves are significantly inhibited. Under the definite concentration scope, an increase in U0126 concentration increases the inhibition degree of conidial germination and appressorium production, but the inhibition degree decreases with elongation of treatment time. The conidial germination, appressorium production, and pathogenicity of S. turcica on susceptible corn leaves are regulated by the MAPK pathway inhibited by U0126.

关键词： Setosphaeria turcica MAPK U0126 conidial germination appressorium production pathogenicity

StKU80, a component in the NHEJ repair pathway, is involved in mycelial morphogenesis, conidiation, appressorium development, and oxidative stress reactions in Exserohilum turcicum

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Journal of Integrative Agriculture 2021年第1期20卷 147-158页

作者： GONG Xiao-dong LIU Yu-wei BI Huan-huan YANG Xiao-rong HAN Jian-min DONG Jin-gao gu shou-qin State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology College of Life SciencesHebei Agricultural UniversityBaoding 071001P.R.China College of Plant Protection Hebei Agricultural UniversityBaoding 071001P.R.China Botou Vocational College Cangzhou 062150P.R.China

Homologous recombination(HR) and nonhomologous end joining(NHEJ) are considered the two main double-strand break(DSB) repair approaches in eukaryotes. Inhibiting the activities of the key component in NHEJ commonly enhances the efficiency of targeted gene knockouts or affects growth and development in higher eukaryotes. However, little is known about the roles of the NHEJ pathway in foliar pathogens. Here we identified a gene designated St KU80, which encodes a putative DNA end-binding protein homologous to yeast Ku80, in the foliar pathogen Exserohilum turcicum. Conserved domain analysis showed that the typical domains VWA, Ku78 and Ku-PK-bind are usually present in Ku70/80 proteins in eukaryotes and are also present in St Ku80. Phylogenetic analysis indicated that St Ku80 is most closely related to Ku80(XP001802136.1) from Parastagonospora nodorum, followed by Ku80(AGF90044.1) from Monascus ruber. Furthermore, the gene knockout mutants ΔSt KU80-1 and ΔSt KU80-2 were obtained. These mutants displayed longer septas, thinner cell walls, smaller amounts of substances on cell wall surfaces, and more mitochondria per cell than the wild-type(WT) strain but similar HT-toxin activity. The mutants did not produce conidia and mature appressoria. On the other hand, the mutants were highly sensitive to H2O2, but not to ultraviolet radiation. In summary, the St KU80 plays devious roles in regulating the development of E. turcicum.

关键词： Exserohilum turcicum pathogenicity gene knockout growth and development

MAP kinase gene STK1 is required for hyphal, conidial, and appressorial development, toxin biosynthesis, pathogenicity, and hypertonic stress response in the plant pathogenic fungus Setosphaeria turcica

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Journal of Integrative Agriculture 2016年第12期15卷 2786-2794页

作者： LI Po GONG Xiao-dong JIA Hui FAN Yong-shan ZHANG Yun-feng CAO Zhi-yan HAO Zhi-min HAN Jian-min gu shou-qin DONG Jin-gao Mycotoxin and Molecular Plant Pathology Laboratory Agricultural University of Hebei Baoding 071000 P.R. China Institute of Plant Protection Hebei Academy of Agriculture and Forestry Sciences Baoding 071000 P.R. China Tangshan Normal University Tangshan 063000 P.R. China

The mitogen-activated protein kinase （MAPK）, a key signal transduction component in the MAPK cascade pathway, regulates a variety of physiological activities in eukaryotes. However, little is known of the role MAPK plays in phytopathogenic fungi. In this research, we cloned the MAPK gene STK1 from the northern corn leaf blight pathogen Setosphaeria turcica and found that the gene shared high homology with the high osmolality glycerol （HOG） MAPK gene HOG1 of Saccharomy- ces cerevisiae. In addition, gene knockout technology was employed to investigate the function of STKI. Gene knockout mutants （KOs） were found to have altered hyphae morphology and no conidiogenesis, though they did show similar radial growth rate compared to the wild-type strain （WT）. Furthermore, microscope observations indicated that STK1 KOs did not form normal appressoria at 48 h post-inoculation on a hydrophobic surface. STK1 KOs had reduced virulence, a significantly altered Helminthosporium turcicum （HT）-toxin composition, and diminished pathogenicity on the leaves of susceptible inbred corn OH43. Mycelium morphology appeared to be significantly swollen and the radial growth rates of STK1 KOs declined in comparison with WT under high osmotic stress. These results suggested that STK1 affects the hyphae development, conidiogenesis, and pathogenicity of S. turcica by regulating appressorium development and HT-toxin biosynthesis. Moreover, the gene appears to be involved in the hypertonic stress response in S. turcica.

关键词： Setosphaeria turcica MAPK conidiogenesis HT-toxin pathogenicity

Stk2,a Mitogen-Activated Protein Kinase from Setosphaeria turcica,Specifically Complements the Functions of the Fus3 and Kss1 of Saccharomyces cerevisiae in Filamentation,Invasive Growth,and Mating Behavior

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Journal of Integrative Agriculture 2013年第12期12卷 2209-2216页

作者： gu shou-qin YANG Yang LI Po ZHANG Chang-zhi FAN Yu ZHANG Xiao-yu TIAN Lan HAO Zhi-min CAO Zhi-yan GONG Xiao-dong FAN Yong-shan HAN Jian-min DONG Jin-gao Mycotoxin and Molecular Plant Pathology Laboratory Agricultural University of Hebei Institute of Plant Protection Hebei Academy of Agriculture and Forestry Sciences Tangshan Normal University

Setosphaeria turcica,an essential phytopathogenic fungus,is the primary cause of serious yield losses in corn; however,its pathogenic mechanism is poorly *** cloned STK2,a newly discovered mitogen-activated protein kinase gene with a deduced amino acid sequence that is 96% identical to MAK2 from Phaeosphaeria nodorum,56% identical to KSS1 and 57% identical to FUS3 from Saccharomyces *** deduce Stk2 function in *** and to identify the genetic relationship between STK2 and KSS1/FUS3 from ***,a restructured vector containing the open reading frame of STK2 was transformed into a fus3/kss1 double deletion mutant of *** results show that the STK2 complementary strain clearly formed pseudohyphae and ascospores,and the strain grew on the surface of the medium after rinsing with sterile water and the characteristics of the complementary strain was the same as the wild-type ***,STK2 complemented the function of KSS1 in filamentation and invasive growth,as well as the mating behavior of FUS3 in ***,however,its exact functions in *** will be studied in the future research.

关键词： Setosphaeria turcica MAPK filamentation invasive growth mating behavior

MAPK途径对玉米大斑病菌HT-毒素产生和生物学活性的调控作用

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中国农业科学 2008年第1期41卷 86-92页

作者：范永山谷守芹董金皋董娜河北农业大学真菌毒素实验室河北农业大学真菌毒素实验室河北保定071001 唐山师范学院生命科学系河北唐山063000

【目的】通过研究促分裂原活化蛋白酶(MAPK)途径对玉米大斑病菌HT-毒素产生及生物学活性的影响,探索该途径对玉米大斑病菌致病性的调控机制。【方法】利用MAPK途径特异性抑制剂U0126分析该途径对玉米大斑病菌HT-毒素产生及活性的影响。... 详细信息

【目的】通过研究促分裂原活化蛋白酶(MAPK)途径对玉米大斑病菌HT-毒素产生及生物学活性的影响,探索该途径对玉米大斑病菌致病性的调控机制。【方法】利用MAPK途径特异性抑制剂U0126分析该途径对玉米大斑病菌HT-毒素产生及活性的影响。【结果】1～20μmol·L-1 U0126处理后,菌株01-23的HT-毒素组成和含量均发生了变化,部分组分的合成受到抑制,并诱导产生了一些新的毒素组分。1～20μmol·L-1 U0126处理后获得的HT-毒素在感病寄主B37和B37Ht1玉米自交系上的生物学活性都受到了显著抑制,在B37玉米叶片上的抑制程度强于B37Ht1。随着U0126浓度的增加,菌株01-23的HT-毒素在玉米叶片上产生的坏死斑面积减小,抑制程度增加,但不能完全抑制病斑产生,10μmol·L-1 U0126处理和20μmol·L-1 U0126处理间差异不显著。【结论】MAPK信号转导途径对玉米大斑病菌HT-毒素的产生和生物学活性具有一定的调控作用,但HT-毒素的产生和活性还受其它因素影响,具有更复杂的调控机制。

关键词：玉米大斑病菌促分裂原活化蛋白酶 HT-毒素 U0126

特异性MEK抑制剂U0126对玉米大斑病菌孢子萌发、附着胞产生和致病性的影响

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中国农业科学 2006年第1期39卷 66-73页

作者：范永山谷守芹董金皋董秉芳唐山063000 河北农业大学真菌毒素实验室河北农业大学真菌毒素实验室保定071001唐山师范学院生物科学技术系

目的研究MAPK信号转导途径对玉米大斑病菌生长、发育和致病性的调控作用,为明确玉米大斑病菌和玉米之间互作的分子机制奠定基础,对玉米大斑病的有效防治也有重要的理论意义。方法利用MEK特异性抑制剂U0126处理玉米大斑病菌,观测该抑制... 详细信息

目的研究MAPK信号转导途径对玉米大斑病菌生长、发育和致病性的调控作用,为明确玉米大斑病菌和玉米之间互作的分子机制奠定基础,对玉米大斑病的有效防治也有重要的理论意义。方法利用MEK特异性抑制剂U0126处理玉米大斑病菌,观测该抑制剂对玉米大斑病菌孢子萌发、附着胞发育和致病性的影响。结果U0126对玉米大斑的菌落形态和生长速度没有显著影响,可以形成正常的菌丝、分生孢子,但分生孢子萌发时间晚,芽管短,分生孢子萌发百分率和附着胞产生数目下降,玉米叶片的发病时间延迟2～3d,发病率下降30%左右。在一定浓度范围内,U0126对分生孢子萌发和附着胞产生的抑制程度随着浓度增加而上升,但随着处理时间的延长而下降。结论玉米大斑病菌的分生孢子萌发、附着胞产生和对感病玉米叶片的致病能力受U0126抑制的MAPK信号转导途径调节。

关键词：玉米大斑病菌 MAPK U0126 分生孢子萌发附着胞产生致病性

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外源水杨酸对玉米大斑病菌生长发育的影响

华北农学报 2010年第2期25卷 229-232页

作者：谷守芹张运峰范永山路粉河北农业大学生命科学学院河北保定071000 唐山师范学院河北唐山063000

以玉米大斑病菌野生型菌株01-23为供试菌株,观测不同浓度水杨酸处理对病菌分生孢子萌发、菌落生长速度及形态、菌丝发育等方面的影响。结果表明,30μg/mL水杨酸处理对病菌分生孢子萌发有显著的促进作用,处理后24h孢子萌发率是对照的1.5... 详细信息

以玉米大斑病菌野生型菌株01-23为供试菌株,观测不同浓度水杨酸处理对病菌分生孢子萌发、菌落生长速度及形态、菌丝发育等方面的影响。结果表明,30μg/mL水杨酸处理对病菌分生孢子萌发有显著的促进作用,处理后24h孢子萌发率是对照的1.53倍。不同浓度水杨酸处理对病菌菌落生长速度也有明显促进作用:15μg/mL水杨酸处理,第8天菌落的生长速度提高了86%;30μg/mL水杨酸处理,第8天菌落的生长速度提高了61%;60μg/mL水杨酸处理,第10天菌落生长速度提高了59%。不同浓度水杨酸处理对菌丝发育也有显著影响:水杨酸浓度为15μg/mL时,菌丝发育没有明显变化;水杨酸浓度为30μg/mL和60μg/mL时,菌丝直径及菌丝形态也无明显变化,但部分菌丝中致密的内含物减少,出现了较分散的空泡状结构,且畸形菌丝的数目增多。水杨酸处理对分生孢子的形态、产生时间及菌落颜色均无明显的影响。

关键词：玉米大斑病菌水杨酸生长发育

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玉米抗大斑病相关基因片段的表达研究

华北农学报 2008年第4期23卷 85-88页

作者：郝敏谷守芹张莹董金皋韩建民河北北方学院医学技术学院河北张家口075000 河北农业大学真菌毒素实验室河北保定071001

以玉米自交系B37Ht1与玉米大斑病菌0号小种和1号小种构成非亲和性互作和亲和性互作体系,利用mRNA差异显示(DDRT-PCR)技术研究了互作过程中玉米叶片基因表达的情况。结果表明,非亲和性互作中特有的表达片段7条,较亲和性互作及对照表达明... 详细信息

以玉米自交系B37Ht1与玉米大斑病菌0号小种和1号小种构成非亲和性互作和亲和性互作体系,利用mRNA差异显示(DDRT-PCR)技术研究了互作过程中玉米叶片基因表达的情况。结果表明,非亲和性互作中特有的表达片段7条,较亲和性互作及对照表达明显增强的片段21条,表达明显减弱的18条。对非亲和性互作中特有的表达片段进行克隆、测序及同源性分析,发现它们与已知抗病基因没有同源性。

关键词：玉米玉米大斑病 mRNA差异显示抗病相关基因片段

利用mRNA差异显示技术分离和克隆玉米抗大斑病相关基因片段

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植物病理学报 2005年第5期35卷 463-465页

作者：谷守芹范永山韩建民董金皋河北农业大学真菌毒素实验室保定071000

玉米大斑病是由玉米大斑病菌（Setosphaeria mrcica)引起的玉米叶部主要病害。在玉米的搞病反应中玉米与大斑病菌间存在着复杂的互作关系。要阐明玉米与搞病过程中抗病基因、参与信号转导基……

关键词： mRNA差异显示技术玉米大斑病菌基因片段技术分离克隆抗病反应抗病相关基因抗病过程基因表达互作关系

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热激对玉米大斑病菌生长和发育的影响

玉米科学 2010年第4期18卷 136-138,141页

作者：谷守芹张运峰范永山路粉河北农业大学生命科学学院河北保定071000 唐山师范学院河北唐山063000

通过对玉米大斑病菌进行热激处理,观测热激对病菌孢子萌发、菌丝生长速度、菌落形态、菌丝形态等方面的影响。结果表明:热激处理对分生孢子萌发率的影响与热激温度有关,42℃热激促进孢子萌发,45℃热激对孢子萌发没有显著影响,48℃热激... 详细信息

通过对玉米大斑病菌进行热激处理,观测热激对病菌孢子萌发、菌丝生长速度、菌落形态、菌丝形态等方面的影响。结果表明:热激处理对分生孢子萌发率的影响与热激温度有关,42℃热激促进孢子萌发,45℃热激对孢子萌发没有显著影响,48℃热激则会显著抑制孢子萌发;42℃～48℃热激处理会降低病菌的菌落生长速度,热激温度越高菌落生长速度越慢;42℃～48℃热激对菌丝形态、分生孢子形态、菌落颜色和菌落形态均无明显影响,但菌丝细胞中的内容物减少,有时出现内容物缺失的菌丝细胞。

关键词：玉米大斑病菌热激生长发育