Abstract
While the functions of vacuolar processing enzymes (VPE) in plant responses to pathogens are now better established, their possible roles against insect herbivores need validation. Inspired by an observation that βVPE and δVPE represent the most highly upregulated transcripts in leaves of Solanum bulbocastanum and its backcross progenies that were highly resistant to the phloem-feeding green peach aphids (Myzus persicae), we used a reverse genetic approach in Arabidopsis to investigate whether T-DNA insertion mutants for specific βVPE and δVPE loci would exhibit compromised defenses at the whole-plant, physiological, and molecular levels. We hypothesized that impaired expression of VPE genes would lead to hyper-susceptibility if VPE-mediated processes were essential for basal defenses, given that Arabidopsis is naturally susceptible. Compared to the wild-type, βvpe and δvpe mutants exhibited an early onset of leaf curling and necrosis, rapid aphid population build-up, and significant physiological tissue injuries as a result of herbivory. This indicates that basal defenses were compromised by the loss-of-functions of either VPE-encoding gene. VPE co-expression networks were comprised of genes involved in sucrose metabolism and transport (AtSUC5, AtSUS3, and invertases), efflux and oxidative detoxification proteins, and modulators of gene-for-gene-type disease response linked to sucrose metabolism (AtSWEET13 and AtSWEET15). These results suggest that VPEs play a role in basal defenses by mediating mechanisms of detoxification and metabolite availability in host tissues to counteract the rapid depletion caused by the phloem sucker. These processes might be important as first line of defense to delay the progression of perturbations caused by herbivory.
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Acknowledgements
The authors thank Myles Butler, Jason Cooney, and Siobhan Cusack for their technical assistance to this project. Promoter sequence analysis was performed using the supercomputing facilities at the National Institute of Genetics, Mishima, Japan. This project was supported by the Maine Agricultural and Forest Experiment Station, University of Maine (Publication 3412) and the USDA National Institute of Food and Agriculture.
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Fig. S1. Putative orthologs of βVPE and δVPE of Arabidopsis were among the most abundant transcripts in a suppression subtractive hybridization library obtained from the leaves of the aphid-resistant wild species of potato S. bulbocastanum (Acc. 243510) infested with M. persicae (data not shown). Differential expression of the putative βVPE and δVPE genes was confirmed by quantitative real-time PCR analysis across a comparison panel that included the resistant donor parent S. bulbocastanum (Sbu243510), susceptible recurrent parent S. tuberosum cv. Ivory Crisp, and two of their contrasting sibling backcross derivatives (Sbu8.5 = resistant; Sbu8.15 = susceptible). Backcross lines were derived from a somatic hybrid between S. tuberosum and S. bulbocastanum backcrossed for four generations to Ivory Crisp. Resistant Sbu8.5 exhibited the βVPE (A) and δVPE (B) expression profile of the donor parent Sbu243510. Susceptible Sbu8.15 exhibited the βVPE and δVPE expression profile of the recurrent parent Ivory Crisp. Relative expression values represent the mean + standard error (n = 3).
Fig. S2. Relative expression of βVPE and δVPE in the leaves of WT Arabidopsis plants in response to aphids (M. persicae) and other factors reported to affect their expression such as Sclerotinia sclerotiorum (pathogen) and various chemical signals such as salicylic acid (SA) and methyl jasmonate (JA). Relative expression values represent the mean + standard error (n = 4).
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Alpuerto, J.B., Mukherjee, A., Kitazumi, A. et al. Impaired expressions of the beta and delta isoforms of vacuolar processing enzymes compromise the basal defenses of Arabidopsis thaliana against the phloem-feeding insect Myzus persicae . Acta Physiol Plant 39, 233 (2017). https://doi.org/10.1007/s11738-017-2529-z
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DOI: https://doi.org/10.1007/s11738-017-2529-z