This greenhouse study evaluated the effects of two chemical inducers for priming kidney bean seedlings against a bacterial wilt disease. This study's central premise was that chlorine dioxide's oxidant properties would mimic the signaling properties of radical oxygen species, thereby initiating a cascade of molecular plant defenses, including the synthesis of salicylic acid (SA). This signaling agent then initiates a cascade of pre-defense activities to provide a more rapid and robust natural defense against pathogen attacks. This factorial study included two levels for a foliar chlorine dioxide treatment and two for a bacterial wilt inoculation treatment. The two plant response variables were free and conjugated salicylic acid levels sampled in leaf tissue over two collection dates. Half of the 96 plants were inoculated with a bacterial culture that causes common bean wilt disease. Leaf tissue was harvested 17 to 32 h and 960 h after the wilt inoculation to determine the temporal dynamics of SA due to chemical treatments. Also, PCR tests were used to verify wilt presence in the inoculated plants. Inoculation of the wilt disease did not affect free SA when leaf tissue was sampled from 17 to 32 h. after wilt inoculation. However, chlorine dioxide applied at 400 mg/l and sampled at 20 h after inoculation resulted in a 15-fold increase in free SA over the control. Also, chlorine dioxide applied at 400 mg/l with leaf tissue sampled at 26 h after inoculation resulted in a 33-fold increase in conjugate SA levels compared to the control plants. Leaf tissue sampled at 960 h after the inoculation showed no free SA differences among the chemical treatments. However, the inoculated plant had a 15.9-fold increase in free SA compared to the non-inoculated plants. The priming effect on kidney bean seedlings using a single chlorine dioxide foliage application temporarily increased free and conjugate SA. The free and conjugate SA levels for the non-inoculated plants returned to baseline levels when sampled at 960 h. These results indicate that primed plants elevate SA up to several weeks with a slow decline back to baseline levels. Stem injection of the bacterial wilt bypassed the immunity mechanisms present in leaves, which significantly increased the wilt injury levels. Stem injection negated much of the foliar defenses, which overshadowed the priming effects of the chemical treatments on plant immunity and foliar defenses. The second leaf sampling on newly formed leaves reveals elevated SA levels in the inoculated plants but not in the non-inoculated plants.

A field study was conducted at the National Ornamental Research Site at Dominican University California (NORS-DUC). The study goal was to evaluate three chemical inducers applied as foliar treatments for controlling Phytophthora ramorum, on Rhododendron x ‘Cunningham’s White’ nursery plants. The inducers were chlorine dioxide (ElectroBiocide), hydrogen peroxide (OxiDate 2.0), and acibenzolar-s methyl (Actigard). Water samples from the electrostatic sprayer were measured for three physicochemical water properties. Visual assessment of plant foliage, based on the Horsfall- Barratt scale, was conducted at three and five months after chemical treatments. Foliar fluorescence (Fv/Fm) was measured over three dates. The success of P. ramorum inoculations were determined using qPCR methods. Visual assessment across both months showed no signs of P. ramorum infection or chemical injury symptoms. However, P. ramorum infection vis-à-vis qPCR analysis was confirmed. The September Fv/Fm results revealed that all the chemical inducer treatments were equivalent to the water treatment, except for Actigard. The qPCR results were in general agreement with the Fv/Fm results indicating that the rhododendrons were successfully inoculated with P. ramorum but were non-symptomatic. The electrostatic sprayer ionized the water droplets, resulting in increased Fv/Fm values for the water treatments 90 days after application. There was a three-month delay in fluorescence responses to the most effective chemical applications, indicating that woody plants may need to be monitored over the long term to determine accurate responses to foliar treatments.

This greenhouse study evaluated the effects of two chemical primers for kidney bean seedlings against a bacterial wilt (Curtobacterium flaccumfaciens pv. Flaccumfaciens) (CFF). The premise of this study was that the oxidant primers would mimic the signaling properties of radical oxygen species and initiate a cascade of molecular defenses. The factorial study included two levels for the foliar chlorine dioxide treatment, and two levels for the bacterial wilt inoculation treatment, plus two supplemental chemical treatments. The foliage response variables were gas exchange and fluorescence. There was a 36, 154, and 70% reduction in Pn, gs, and E, respectively, at 39 DAT when comparing the inoculated control to the non-inoculated control. The chlorine dioxide primers lowered leaf temperatures and leaf vapor pressure deficit in the CFF wilt inoculated plants. The chlorine dioxide primers improved gas exchange at 39 DAT when compared to the water treatments. Part 1 and 2 of this series conclude that the chlorine dioxide primers can activate a long-term, systemic acquired resistance (SAR) response in kidney bean plants infected with the CFF wilt. The Part 2 article also concludes that the EB treatments caused several inexplicable correlations among the gas exchange responses. A structured water premise was proposed as an explanation for the gas exchange anomalies due to the EB treatments. Intuitively, this study suggests that chlorine dioxide primers can initiate a series of ROS and salicylic acid signals that activate a suite of mechanisms that provide universal, multifaceted plant immunity that is sustained across a crop season.