A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and RsJMT) and salicylic acid (SA) (RsPAL and RsICS). Furthermore, RsPPO, a putative marker gene for oxidative stress response was successfully cloned from R. simsii. A CTAB-based extraction protocol was optimized to assure excellent RNA quality for subsequent RT-qPCR analysis. The RT-qPCR protocol was extensively tested and RsRG7 and RsRG14 were selected as reference genes from a geNorm pilot study. Validation of the marker genes was done after application with elicitors [methyl jasmonate (MeJA), coronatine, beta-aminobutyric acid and acibenzolar-Smethyl] or wounding. Both 100 mu M MeJA and 0.1 lM coronatine had a significant effect on the expression of all marker genes. Foliar application of MeJA on the shoots resulted in a significantly earlier response when compared to root application and subsequent sampling of the shoots. Expression patterns after MeJA treatment were generally the same in six R. simsii genotypes: 'Nordlicht', 'Elien', 'Aiko Pink', ' Michelle Marie', 'Mevrouw Gerard Kint' and 'Sachsenstern'. Wounding resulted in the same expression patterns as MeJA treatment except for RsJMT. None of the genotypes showed a significant induction of the latter gene 6 h upon wounding. Findings of these experiments indicated that the tolerant genotype 'Elien' has low basal expression levels of RsPPO. This might be the first step towards the breeding of mite-tolerant genotypes

Broad mite Polyphagotarsonemus latus, is a key pest of pot azalea (Rhododendron simsii Planch hybrids). No information on variation in susceptibility or potential tolerance within these plants is available to date. We evaluated a selection of 32 Rhododendron cultivars, mainly R. simsii hybrids, for susceptibility to broad mite. The plants were artificially infested in a greenhouse by surrounding each azalea with four broad mite-infested English ivy plants (Hedera helix). Broad mite infestation was evaluated by counting the number of broad mites per shoot tip and assigning a damage rate. Results indicated a comparable infestation rate expressed as the number of mites on all cultivars at the initial stage of the experiments. Correlations between the average damage rate and the number of broad mites per shoot tip on all cultivars at different time intervals were significantly positive, although low, in each experiment, with R 2 -values of 0.14 and 0.61. At the end of the experiments significant differences in susceptibility between the evaluated cultivars were observed. The cultivars, 'Emil De Coninck' and 'Mont Blanc' were rated as the most susceptible, whereas 'Mistral' and its bud sport 'Elien' were tolerant towards the broad mite, because damage rates were low and very few broad mites were found.

The broad mite, Polyphagotarsonemus latus (Banks), is one of the major pests causing severe economic damage in Rhododendron simsii Planch hybrid production in Belgium. In order to optimize biological control programs and to parameterize warning programs, we studied the effect of environmental temperature on the development of P. latus on R. simsii leaves. In combination with a photoperiod of 16:8 h (L:D) and a relative humidity of 80 ± 5 %, six constant temperatures (15, 17, 20, 25, 30 and 33 ± 1 °C), were studied. Total developmental times of 13.3, 10.5, 6.6, 4.2, 3.5 and 4.0 days were measured, respective to each of the aforementioned temperatures. Development of females took significantly longer than that of males at 15, 17, 20 and 30 °C. Survival rates observed between 17 and 30 °C varied between 43.5 and 96.9 %. Lower survival rates were found at 15 and 33 °C, i.e. 31.8 and 23.6 %, respectively. The lower, optimal and upper developmental threshold (t min , t opt and t max , respectively) and thermal constant (K) of the pest were estimated for each life stage by a linear and two non-linear models. Based on measurements of total development of P. latus thermal thresholds of 10.0, 30.1 and 36.0 °C were calculated for t min , t opt and t max , respectively. The number of degree-days needed to complete immature development when feeding on R. simsii was 66.7.

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