小麦赤霉病菌FgVPS26与FgRab7的相互作用及对DON毒素合成的影响
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摘要
小麦赤霉病菌合成DON毒素是多基因控制的次级代谢过程,阐明各种基因的调控作用有利于进行科学防控。以赤霉病菌为研究对象,通过酵母双杂交、实时定量PCR及ELISA等技术研究FgVPS26与FgRab7的相互作用及对DON毒素生物合成的影响。结果表明:FgVPS26和激活态FgRab7可以相互作用,但与原始态FgRab7不能相互合作。FgVPS26和FgRab7基因可相互影响其表达,与野生型菌株PH-1相比,当FgVPS26和FgRab7其中一个基因缺失时另一个基因的表达量将显著下降。另外,FgVPS26或FgRab7基因缺失也显著降低与DON毒素合成相关的Tri基因表达,从而导致同一生长时期突变菌株的DON毒素合成量比野生型菌株PH-1明显减少。因此,小麦赤霉病菌中FgVPS26和FgRab7可以相互作用并调控DON毒素的生物合成。
The biosynthesis of deoxynivalenol(DON) toxin is a secondary metabolic process regulated by multiple genes in Fusarium graminearum,and elucidating the regulatory roles of the related genes is conducive to the development of scientific prevention and control measures. In this study,yeast two-hybrid,real-time quantitative PCR and ELISA were employed to investigate the interaction between Fg VPS26 and Fg Rab7 and their effect on the biosynthesis of DON toxin. The results showed that Fg VPS26 could interact with activated Fg Rab7,but could not interact with the original state of Fg Rab7. The expression of Fg VPS26 and Fg Rab7 genes could influence each other,and when one of the genes(Fg VPS26 and Fg Rab7) was knocked out,the expression of the other gene decreased significantly compared with the level in wild-type strain PH-1. Additionally,deletion of Fg VPS26 or Fg Rab7 gene also significantly affected the expression of Tri gene involved in DON toxin synthesis,which resulted in the decrease of DON toxin synthesis of mutant strains compared with that of wild type strain at the same growth stage. Our results suggested that Fg VPS26 and Fg Rab7 could interact with each other and regulate the biosynthesis of DON toxin in Fusarium graminearum.
The biosynthesis of deoxynivalenol(DON) toxin is a secondary metabolic process regulated by multiple genes in Fusarium graminearum,and elucidating the regulatory roles of the related genes is conducive to the development of scientific prevention and control measures. In this study,yeast two-hybrid,real-time quantitative PCR and ELISA were employed to investigate the interaction between Fg VPS26 and Fg Rab7 and their effect on the biosynthesis of DON toxin. The results showed that Fg VPS26 could interact with activated Fg Rab7,but could not interact with the original state of Fg Rab7. The expression of Fg VPS26 and Fg Rab7 genes could influence each other,and when one of the genes(Fg VPS26 and Fg Rab7) was knocked out,the expression of the other gene decreased significantly compared with the level in wild-type strain PH-1. Additionally,deletion of Fg VPS26 or Fg Rab7 gene also significantly affected the expression of Tri gene involved in DON toxin synthesis,which resulted in the decrease of DON toxin synthesis of mutant strains compared with that of wild type strain at the same growth stage. Our results suggested that Fg VPS26 and Fg Rab7 could interact with each other and regulate the biosynthesis of DON toxin in Fusarium graminearum.
引文
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[14]王亚君,翟焕趁,张帅兵,等.小麦赤霉病菌FgRab7调控Tri基因表达及DON毒素合成[J].河南工业大学学报(自然科学版),2017,38(2):33-39.
[15] WANG T,MING Z,WU X C,et al. Rab7:role of its protein interaction cascades in endolysosomal traffic[J]. Cellular Signalling,2011,23(3):516-521.
[16] ZHENG W H,ZHENG H W,ZHAO X,et al.Retrograde trafficking from the endosome to the trans-Golgi network mediated by the retromer is required for fungal development and pathogenicity in Fusarium graminearum[J]. New Phytologist,2016,210(4):1327-1343.
[17] BALDERHAAR H J,ARLT H,OSTROWICZ C,et al. The Rab GTPase Ypt7 is linked to retromer-mediated receptor recycling and fusion at the yeast late endosome[J]. Journal of Cell Science,2010,123(23):4085-4094.
[2] PALAZZINI J,FUMERO V,YERKOVICH N,et al. Correlation between Fusarium graminearum and deoxynivalenol during the 2012/13 wheat Fusarium head blight outbreak in Argentina[J].Cereal Research Communications,2015,43(4):627-637.
[3] RAAD F,NASREDDINE L,HILAN C,et al. Dietary exposure to aflatoxins,ochratoxin A and deoxynivalenol from a total diet study in an adult urban Lebanese population[J]. Food and Chemical Toxicology,2014,73:35-43.
[4]蔡静平,刘新影,翟焕趁.禾谷镰刀菌DON毒素生物合成调控研究进展[J].河南工业大学学报(自然科学版),2016,37(1):114-119.
[5] MARROQU魱N-CARDONA A G,JOHNSON N M,PHILLIPS T D,et al. Mycotoxins in a changing global environment a review[J]. Food and Chemical Toxicology,2014,69:220-230.
[6] NASMITH C G,WALKOWIAK S,WANG L,et al. Tri6 is a global transcription regulator in the phytopathogen Fusarium graminearum[J].PLoS Pathogen,2011,7(9):e1002266.
[7] ZHANG C K,WANG Y,WANG J Q,et al. Functional characterization of Rho family small GTPases in Fusarium graminearum[J]. Fungal Genetics and Biology,2013,61(12):90-99.
[8] YU F W,GU Q,YUN Y Z,et al. The TOR signaling pathway regulates vegetative development and virulence in Fusarium graminearum[J]. New Phytologist,2014,203(1):219-232.
[9] QIN J,WANG G H,JIANG C,et al. Fgk3 glycogen synthase kinase is important for development,pathogenesis,and stress responses in Fusarium graminearum[J]. Scientific Reports,2015,5:8504.
[10] LI Y,LI B,LIU L,et al. FgMon1,a guanine nucleotide exchange factor of FgRab7,is important for vacuole fusion,autophagy and plant infection in Fusarium graminearum[J]. Scientific Reports,2015,5:18101.
[11] CHEN D P,WANG Y,ZHOU X Y,et al. The Sch9 kinase regulates conidium size,stress responses,and pathogenesis in Fusarium graminearum[J]. PLoS One,2014,9(8):e105811.
[12] QI P F,ZHANG Y Z,LIU C H,et al. Fusarium graminearum ATP-Binding Cassette transporter gene FgABCC9 is required for its transportation of salicylic acid,fungicide resistance,Mycelial growth and pathogenicity towards wheat[J]. International Journal of Molecular Sciences,2018,19:2351.
[13] ZHENG H W,LI L P,MIAO P F,et al.FgSec2A,a guanine nucleotide exchange factor of FgRab8,is important for polarized growth,pathogenicity and DON production in Fusarium graminearum[J]. Environmental Microbiology,2018,20(9):3378-3392.
[14]王亚君,翟焕趁,张帅兵,等.小麦赤霉病菌FgRab7调控Tri基因表达及DON毒素合成[J].河南工业大学学报(自然科学版),2017,38(2):33-39.
[15] WANG T,MING Z,WU X C,et al. Rab7:role of its protein interaction cascades in endolysosomal traffic[J]. Cellular Signalling,2011,23(3):516-521.
[16] ZHENG W H,ZHENG H W,ZHAO X,et al.Retrograde trafficking from the endosome to the trans-Golgi network mediated by the retromer is required for fungal development and pathogenicity in Fusarium graminearum[J]. New Phytologist,2016,210(4):1327-1343.
[17] BALDERHAAR H J,ARLT H,OSTROWICZ C,et al. The Rab GTPase Ypt7 is linked to retromer-mediated receptor recycling and fusion at the yeast late endosome[J]. Journal of Cell Science,2010,123(23):4085-4094.