Effect of Bacillus subtilis and chitosan applications on green mold (Penicilium digitatum Sacc.) decay in citrus fruit

Abstract The suppression of green mold postharvest caused by Penicilium digitatum , was evaluated in mandarin fruit after application of Bacillus subtilis ABS-S14 endospores, a crude extract from its culture medium, cyclic lipopeptide antibiotics (CLPs) and chitosan. Their abilities to elicit the accumulation of the plant defense related enzymes; peroxidase (POX) and l -phenylalanine ammonia-lyase (PAL) were also tested. Both B. subtilis ABS-S14 endospores and a crude extract from its growth medium showed strong antagonistic activities against P. digitatum . The HPLC profiles of the crude extract indicated that the antifungal compounds produced by this strain of B. subtilis consisted of three clusters of the iturin A, fengycin and surfactin families of CLPs. The in vitro and in vivo inhibitory effects of the partially purified iturins and fengycins on retardation of the growth of P. digitatum were demonstrated, whereas the surfactins had no direct effect. Efficacy tests, using bacterial endospores, the crude extract and chitosan showed a significant reduction of fruit decay compared to those found in the presence of individual CLPs, and it was clearly demonstrated that the B. subtilis ABS-S14, itself and its crude extract and chitosan each induced the activities of POX and PAL in the infected flavedo tissues of mandarin fruit. They also co-enhanced the protection of fruit from the green mold pathogen P. digitatum .

[1]  Lluís Palou,et al.  Alternatives to conventional fungicides for the control of citrus postharvest green and blue moulds , 2008 .

[2]  Hong-xia Liu,et al.  Analysis of defence enzymes induced by antagonistic bacteriumBacillus subtilis strain AR12 towardsRalstonia solanacearum in tomato , 2008, Annals of Microbiology.

[3]  Samir Droby,et al.  Twenty years of postharvest biocontrol research: Is it time for a new paradigm? , 2009 .

[4]  Jun Liu,et al.  Functions of Lipopeptides Bacillomycin D and Fengycin in Antagonism of Bacillus amyloliquefaciens C06 towards Monilinia fructicola , 2011, Journal of Molecular Microbiology and Biotechnology.

[5]  Z. Yang,et al.  Fengycin antibiotics isolated from B-FS01 culture inhibit the growth of Fusarium moniliforme Sheldon ATCC 38932. , 2007, FEMS microbiology letters.

[6]  C. Pérez.  Antibiotic assay by agar-well diffusion method , 1990 .

[7]  Yong Xu,et al.  Effects of chitosan on control of postharvest diseases and physiological responses of tomato fruit , 2007 .

[8]  Yu Zheng,et al.  Quantitative changes of plant defense enzymes and phytohormone in biocontrol of cucumber Fusarium wilt by Bacillus subtilis B579 , 2010 .

[9]  A. Gamliel,et al.  Toxicity of chloronitrobenzenes toFusarium oxysporum and rhizoctonia solani as related to their structure , 1989, Phytoparasitica.

[10]  W. Leelasuphakul,et al.  Growth inhibitory properties of Bacillus subtilis strains and their metabolites against the green mold pathogen (Penicillium digitatum Sacc.) of citrus fruit , 2008 .

[11]  J. Kennedy,et al.  Effects of chitosan and oligochitosan on growth of two fungal pathogens and physiological properties in pear fruit , 2010 .

[12]  Ting Yu,et al.  Synergistic effect of chitosan and Cryptococcus laurentii on inhibition of Penicillium expansum infections. , 2007, International journal of food microbiology.

[13]  Renping Li,et al.  Effects of Chitin and Its Derivative Chitosan on Postharvest Decay of Fruits: A Review , 2011, International journal of molecular sciences.

[14]  S. Sangchote,et al.  Biocontrol of citrus green mould and postharvest quality parameters. , 2013 .

[15]  F. Nigro,et al.  Effects of Pre‐ and Postharvest Chitosan Treatments to Control Storage Grey Mold of Table Grapes , 2002 .

[16]  M. Shoda,et al.  Biocontrol of Rhizoctonia solani Damping-Off of Tomato with Bacillus subtilis RB14 , 1996, Applied and environmental microbiology.

[17]  C. Casals,et al.  The combination of curing with either chitosan or Bacillus subtilis CPA-8 to control brown rot infections caused by Monilinia fructicola , 2012 .

[18]  S. Phongpaichit,et al.  Purification, characterization and synergistic activity of β-1,3-glucanase and antibiotic extract from an antagonistic Bacillus subtilis NSRS 89-24 against rice blast and sheath blight , 2006 .

[19]  R. Zeng,et al.  Physiological and cytological mechanisms of silicon-induced resistance in rice against blast disease. , 2008, Physiologia plantarum.

[20]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[21]  W. Liang,et al.  Effect of Crude Extract from Bacillus Subtilis LB5 Cultivated Broth on Conidial Germination of Colletotrichum Gloeosporioides , 2012 .

[22]  O. Kuipers,et al.  The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca. , 2007, Molecular plant-microbe interactions : MPMI.

[23]  S. F. Pascholati,et al.  Chitosan reduces infection by Guignardia citricarpa in postharvest 'Valencia' oranges , 2009 .

[24]  Luis González-Candelas,et al.  Spatial study of antioxidant enzymes, peroxidase and phenylalanine ammonia-lyase in the citrus fruit–Penicillium digitatum interaction , 2006 .

[25]  O. Nybroe,et al.  Natural functions of lipopeptides from Bacillus and Pseudomonas: more than surfactants and antibiotics. , 2010, FEMS microbiology reviews.

[26]  S. Chulze,et al.  Biocontrol of Bacillus subtilis against Fusarium verticillioides in vitro and at the maize root level. , 2005, Research in microbiology.

[27]  D. Haas,et al.  Biological control of soil-borne pathogens by fluorescent pseudomonads , 2005, Nature Reviews Microbiology.

[28]  C. Rodriguez,et al.  An antibiotic assay by the agar well diffusion method , 1990 .

[29]  A. Mounteer,et al.  Rhizobacterial induction of systemic resistance in tomato plants: non-specific protection and increase in enzyme activities , 2004 .

[30]  R. Reuveni,et al.  A Rapid Assay for Monitoring Peroxidase Activity in Melon as a Marker for Resistance to Pseudoperonospora cubensis , 1990 .

[31]  M. Ongena,et al.  Bacillus lipopeptides: versatile weapons for plant disease biocontrol. , 2008, Trends in microbiology.

[32]  F. Daayf,et al.  Treatment of chickpea with Rhizobium isolates enhances the expression of phenylpropanoid defense-related genes in response to infection by Fusarium oxysporum f. sp. ciceris. , 2007, Plant physiology and biochemistry : PPB.

[33]  A. de Vicente,et al.  Effect of lipopeptides of antagonistic strains of Bacillus subtilis on the morphology and ultrastructure of the cucurbit fungal pathogen Podosphaera fusca , 2007, Journal of applied microbiology.

[34]  L. Korsten,et al.  Effect of volatile compounds produced by Bacillus strains on postharvest decay in citrus , 2010 .