Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (16): 3167-3173.doi: 10.3864/j.issn.0578-1752.2014.16.005

• PLANT PROTECTION • Previous Articles     Next Articles

Mechanism Analysis of Kynurenine 3-monooxygenase Gene BcKMO in Regulation of Pathogenicity in Botrytis cinerea

 LI  Pei-Fen, ZHAO  Fu-Xin, DONG  Li-Ping, ZHENG  Hui-Xin, ZHAO  Bin, HAN  Jian-Min, XING  Ji-Hong, DONG  Jin-Gao   

  1. Agricultural University of Hebei/ Laboratory of Mycotoxin and Molecular Plant Pathology, Baoding 071001, Hebei
  • Received:2014-01-27 Online:2014-08-18 Published:2014-04-15

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Abstract: 【Objective】The objective of this study is to reveal the molecular mechanism of kynurenine 3-monooxygenase gene BcKMO in regulating pathogenicity of Botrytis cinerea and lay a foundation for clarifing the pathogenic mechanism of B. cinerea in the future.【Method】The activities of polymethylgalacturonase (PMG), endopolygalacturonase (PG), cellulase (CX), polygalacturonic acid transeliminase (PGTE) and pectin methyl transelimination enzyme (PMTE) were analyzed in the wild-type strain (WT), the BcKMO gene ATMT mutant (BCG183) and gene complement mutant (BCG183/BcKMO) by DNS and Hoffman methods. The toxin was isolated from WT, BCG183 and BCG183/BcKMO and the activity of toxin was analyzed. Acid production assays was performed on PDA medium with bromothymol blue. Penetrability of WT, BCG183 and BCG183/BcKMO was detected with onion epidermis spread on PDA medium. Real-time PCR was used to measure the transcription levels of pathogenicity-related genes, e.g., Bac, Bcg2, Bcg3, PkaR, Bmp1, Sak1, Bcreg1, Bos1, Bcp1, Ras2, Bcpg1, and Sod1 in WT, BCG183 and BCG183/BcKMO.【Result】The activity of PMG and PG in mutant BCG183 were significantly higher than that of WT and BCG183/BcKMO. The CX, PGTE and PMTE activities were no significant difference while compared with WT and BCG183/BcKMO. The toxin activity of mutant BCG183 was significantly higher than that of WT and BCG183/BcKMO. The acid production of the mutant BCG183 significantly decreased. The penetrability of mutant BCG183 appeared no significant difference compared to WT and BCG183/BcKMO. The expression level of pathogenicity-related genes, i.e., Bac, Bcg2, Bcg3, PkaR, Bmp1, Sak1, Bcreg1, Bos1, Bcp1, Ras2, Bcpg1, and Sod1, was obviously up-regulated in mutant BCG183.【Conclusion】The BcKMO gene is involved in regulating cell wall degradation enzyme activity, toxin activity, acid production, penetrability, and the expression of pathogenicity- related genes in B. cinerea.

Key words: Botrytis cinerea , BcKMO , mutants , pathogenicity

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