Abstract
Solanum lycopersicum L. is affected among other pests and diseases, by the actinomycete Clavibacter michiganensis subsp. michiganensis (Cmm), causing important economic losses worldwide. Antimicrobial peptides (AMPs) are amphipathic cationic oligopeptides with which the development of pathogenic microorganisms has been inhibited. Therefore, in this study, we evaluate antimicrobial activity of mesoporous silica nanoparticles (MSN5.4) loaded with human β-defensin-2 (hβD2) and two mutants (TRX-hβD2-M and hβD2-M) against Cmm. hβD2, TRX-hβD2-M and hβD2-M presented a half-maximum inhibitory concentration (IC50) of 3.64, 1.56 and 6.17 μg/mL, respectively. MSNs had average particle sizes of 140 nm (SEM) and a tunable pore diameter of 4.8 up to 5.4 nm (BJH). AMPs were adsorbed more than 99% into MSN and a first release after 24 h was observed. The MSN loaded with the AMPs inhibited the growth of Cmm in solid and liquid media. It was also determined that MSNs protect AMPs from enzymatic degradation when the MSN/AMPs complexes were exposed to a pepsin treatment. An improved AMP performance was registered when it was adsorbed in the mesoporous matrix. The present study could expand the applications of MSNs loaded with AMPs as a biological control and provide new tools for the management of phytopathogenic microorganisms.
Highlights
One of the most important crops worldwide is tomato, the fruit of S. lycopersicumL., of which just in 2019 production exceeded 180 million tons [1]
Orientation of DNA fragments encoding human β-defensin-2 (hβD2) and hβD2M into the pCri-4a vector was assessed by digestion with the restriction enzymes NcoI and
After cleavage by tobacco etch virus protease (TEV) protease and analyzing the unbound fraction of a second purification step by Ni2+ -IMAC, a band near to 4.6 kDa assigned to hβD2 was observed
Summary
One of the most important crops worldwide is tomato, the fruit of S. lycopersicumL., of which just in 2019 production exceeded 180 million tons [1]. One of the most important crops worldwide is tomato, the fruit of S. lycopersicum. Depending on the growth stage and environmental conditions, this crop can be negatively affected by stress, pests and diseases that cause damage to the stem and fruit, and to the root system of the plant [2,3]. The use of contaminated seeds and asymptomatic transplants causes the propagation of the disease, detection methods are inefficient at differentiating Cmm from non-pathogenic Clavibacter strains [5]. The detection of pathogenic microorganisms by means of antibodies functionalized to nanomaterials [6]. When infection occurs in adult plants it could be asymptomatic, eventually producing marketable fruits [4,7,8].
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