ABSTRACT
6-deoxy-6-amino chitosan (aminochitosan) is a water-soluble chitosan derivative with an additional amine group at the C-6 position. This modification has improved aqueous solubility, in vitro antifungal activity and is hypothesized to have enhanced in vivo antifungal activity compared to native chitosan. Gray mold disease in tomatoes is caused by the fungus, Botrytis cinerea, and poses a severe threat both pre- and post-harvest. To investigate the optimal concentration of aminochitosan and its lower molecular weight fractions for antifungal and priming properties in the tomato/B. cinerea pathosystem, different concentrations of aminochitosan were tested in vitro on B. cinerea growth and sporulation and in vivo as a foliar pre-treatment in tomato leaves. The leaves were monitored for photosynthetic changes using multispectral imaging and hydrogen peroxide accumulation using DAB. Despite batch-to-batch variations in aminochitosan, it displayed significantly greater inhibition of B. cinerea in vitro than native chitosan at a minimum concentration of 1 mg/mL. A concentration-dependent increase in the in vitro antifungal activities was observed for radial growth, sporulation, and germination with maximum in vitro inhibition for all the biopolymer batches and lower MW fractions at 2.5 and 5 mg/mL, respectively. However, the inhibition threshold for aminochitosan was identified as 1 mg/mL for spores germinating in vivo, compared to the 2.5 mg/mL threshold in vitro. The pre-treatment of leaves displayed efficacy in priming direct and systemic resistance to B. cinerea infection at 4, 6 and 30 days post-inoculation by maintaining elevated Fv/Fm activity and chlorophyll content due to a stronger and more rapid elicitation of the defense systems at earlier time points. Moreover, these defense systems appear to be ROS-independent at higher concentrations (1 and 2.5 mg/mL). In addition, aminochitosan accumulates in the cell membrane and therefore acts to increase the membrane permeability of cells after foliar spray. These observations corroborate the notion that aminochitosan biopolymers can exert their effects through both direct mechanisms of action and indirect immunostimulatory mechanisms. The contrast between in vitro and in vivo efficacy highlights the bimodal mechanisms of action of aminochitosan and the advantageous role of primed plant defense systems.
ABSTRACT
Maize (Zea mays L.) is a staple crop providing food security to millions of people in sub Saharan Africa. Fusarium verticillioides, an important fungal pathogen, infects maize causing 'Fusarium Ear Rot' disease, which decreases maize kernel yield and the quality of the crop harvested. Currently, no African maize line is completely resistant to infection by F. verticillioides. This study investigated an African maize line, Zea mays CML144, infected with F. verticillioides. Analysis of morphological characteristics showed significant differences between mock-infected and infected plants. RNA-sequencing (RNA-seq) was conducted on plants 14 days post-inoculation to identify differentially expressed genes (DEGs) involved in F. verticillioides infection. Analysis of RNA-seq data revealed DEGs that were both significantly up- and down-regulated in the infected samples compared to the mock-infected control. The maize TPS1 and cytochrome P450 genes were up-regulated, suggesting that kauralexins were involved in the CML144 defense response. This was substantiated by kauralexin analyses, which showed that kauralexins, belonging to class A and B, accumulated in infected maize tissue. Gene ontology terms relating to response to stimulus, chemical stimulus and carbohydrate metabolic processes were enriched, and the genes belonging to these GO-terms were down-regulated. Quantitative real-time PCR was performed on selected DEGs and measurement of phytoalexin accumulation validated the RNA-seq data and GO-analysis results. A comparison of DEGs from this study to DEGs found in F. verticillioides (ITEM 1744) infected susceptible (CO354) and resistant (CO441) maize genotypes in a previous study, matched 18 DEGs with 17 up-regulated and one down-regulated, respectively. This is the first transcriptomic study on the African maize line, CML144, in response to F. verticillioides infection.
