Modeling the Airborne Inoculum of Polystigma amygdalinum to Optimize Fungicide Programs Against Almond Red Leaf Blotch.

Red leaf blotch (RLB) of almond, caused by the ascomycete Polystigma amygdalinum, is a severe foliar disease endemic in the Mediterranean Basin and Middle East. Airborne ascospores of P. amygdalinum were monitored from 2019 to 2021 in two almond orchards in Lleida, Spain, and a Bayesian beta regression was used to model its seasonal dynamics. The selected model incorporated accumulated degree-days (ADD), ADD considering both vapor pressure deficit and rainfall as fixed effects, and a random effect for the year and location. The performance of the model was evaluated in 2022 to optimize RLB fungicide programs by comparing the use of model predictions and action thresholds with the standard program. Two variants were additionally considered in each program to set the frequency between applications, based on (i) a fixed frequency of 21 days or (ii) specific meteorological criteria (spraying within 7 days after rainfalls greater than 10 mm, with daily mean temperatures between 10 and 20°C, and with a minimum frequency of 21 days between applications). Programs were evaluated in terms of RLB incidence and number of applications. The program based on the model with periodic fungicide applications was similarly effective as the standard program, resulting only in a 2.6% higher RLB incidence but with fewer applications (three to four, compared with seven in the standard program). When setting the frequency between applications by using the meteorological criteria, a higher reduction in the number of applications (two to three) was observed, while RLB incidence increased by roughly 16% in both programs. Therefore, the model developed in this study may represent a valuable tool toward a more sustainable fungicide schedule for the control of almond RLB in northeast Spain.

Susceptibility of Almond (Prunus dulcis) Cultivars to Twig Canker and Shoot Blight Caused by Diaporthe amygdali.

Twenty-five almond cultivars were assessed for susceptibility to Diaporthe amygdali, causal agent of twig canker and shoot blight disease. In laboratory experiments, growing twigs were inoculated with four D. amygdali isolates. Moreover, growing shoots of almond cultivars grafted onto INRA 'GF-677' rootstock were used in 4-year field inoculations with one D. amygdali isolate. In both types of experiments, inoculum consisted of agar plugs with mycelium, which were inserted underneath the bark, and the lesion lengths caused by the fungus were measured. Necrotic lesions were observed in the inoculated almond cultivars in both laboratory and field tests, confirming the susceptibility of all evaluated cultivars to all inoculated isolates of D. amygdali. Cultivars were grouped as susceptible or very susceptible according to a cluster analysis. The relationship between some agronomic traits and cultivar susceptibility was also investigated. Blooming and ripening times were found to be relevant variables explaining cultivar performance related to D. amygdali susceptibility. Late and very late blooming and early and medium ripening cultivars were highly susceptible to D. amygdali. Our results may provide valuable information that could assist in ongoing breeding programs of this crop and in the selection of cultivars for new almond plantations.

Esterase, Glutathione S-Transferase and NADPH-Cytochrome P450 Reductase Activity Evaluation in Cacopsylla pyri L. (Hemiptera: Psyllidae) Individual Adults.

Cacopsylla pyri (L.) (Hemiptera: Psyllidae) is a key pest of pear orchards in Spain. The large number of insecticide treatments necessary for control may be an important contributor to the emergence of resistance. Laboratory toxicity and biochemical assays are necessary to validate the existence of insecticide resistance and establish the underlying mechanisms. All the methodologies developed to evaluate enzyme activity in C. pyri to date have incorporated "pools" of adults to detect minimum activity ranges. In this study, we determined the optimal working conditions for evaluation of the activities of esterase, glutathione S-transferase and NADPH-cytochrome P450 reductase in individual insects via colorimetric methods using a microplate reader. The main factors affecting enzymatic analysis activity, such as enzyme source and substrate concentration, filter wavelength, buffer pH, reaction time and additives, were evaluated for optimization. Determining the frequency of resistant individuals within a population could be used as an indicator for the evolution of insecticide resistance over time. Two laboratory strains, one of them selected with cypermethrin, and two field populations were analyzed for this purpose. The data obtained revealed high values and great variation in the activity ranges of esterase (EST) in the insecticide-selected population as well as in the field populations validating the applied methodology.

Remote Sensing Energy Balance Model for the Assessment of Crop Evapotranspiration and Water Status in an Almond Rootstock Collection.

One of the objectives of many studies conducted by breeding programs is to characterize and select rootstocks well-adapted to drought conditions. In recent years, field high-throughput phenotyping methods have been developed to characterize plant traits and to identify the most water use efficient varieties and rootstocks. However, none of these studies have been able to quantify the behavior of crop evapotranspiration in almond rootstocks under different water regimes. In this study, remote sensing phenotyping methods were used to assess the evapotranspiration of almond cv. "Marinada" grafted onto a rootstock collection. In particular, the two-source energy balance and Shuttleworth and Wallace models were used to, respectively, estimate the actual and potential evapotranspiration of almonds grafted onto 10 rootstock under three different irrigation treatments. For this purpose, three flights were conducted during the 2018 and 2019 growing seasons with an aircraft equipped with a thermal and multispectral camera. Stem water potential (Ψ s t e m ) was also measured concomitant to image acquisition. Biophysical traits of the vegetation were firstly assessed through photogrammetry techniques, spectral vegetation indices and the radiative transfer model PROSAIL. The estimates of canopy height, leaf area index and daily fraction of intercepted radiation had root mean square errors of 0.57 m, 0.24 m m-1 and 0.07%, respectively. Findings of this study showed significant differences between rootstocks in all of the evaluated parameters. Cadaman® and Garnem® had the highest canopy vigor traits, evapotranspiration, Ψ s t e m and kernel yield. In contrast, Rootpac® 20 and Rootpac® R had the lowest values of the same parameters, suggesting that this was due to an incompatibility between plum-almond species or to a lower water absorption capability of the rooting system. Among the rootstocks with medium canopy vigor, Adesoto and IRTA 1 had a lower evapotranspiration than Rootpac® 40 and Ishtara®. Water productivity (WP) (kg kernel/mm water evapotranspired) tended to decrease with Ψ s t e m , mainly in 2018. Cadaman® and Garnem® had the highest WP, followed by INRA GF-677, IRTA 1, IRTA 2, and Rootpac® 40. Despite the low Ψ s t e m of Rootpac® R, the WP of this rootstock was also high.

Pedigree analysis of 220 almond genotypes reveals two world mainstream breeding lines based on only three different cultivars.

Loss of genetic variability is an increasing challenge in tree breeding programs due to the repeated use of a reduced number of founder genotypes. However, in almond, little is known about the genetic variability in current breeding stocks, although several cases of inbreeding depression have been reported. To gain insights into the genetic structure in modern breeding programs worldwide, marker-verified pedigree data of 220 almond cultivars and breeding selections were analyzed. Inbreeding coefficients, pairwise relatedness, and genetic contribution were calculated for these genotypes. The results reveal two mainstream breeding lines based on three cultivars: "Tuono", "Cristomorto", and "Nonpareil". Descendants from "Tuono" or "Cristomorto" number 76 (sharing 34 descendants), while "Nonpareil" has 71 descendants. The mean inbreeding coefficient of the analyzed genotypes was 0.041, with 14 genotypes presenting a high inbreeding coefficient, over 0.250. Breeding programs from France, the USA, and Spain showed inbreeding coefficients of 0.075, 0.070, and 0.037, respectively. According to their genetic contribution, modern cultivars from Israel, France, the USA, Spain, and Australia trace back to a maximum of six main founding genotypes. Among the group of 65 genotypes carrying the Sf allele for self-compatibility, the mean relatedness coefficient was 0.125, with "Tuono" as the main founding genotype (24.7% of total genetic contribution). The results broaden our understanding about the tendencies followed in almond breeding over the last 50 years and will have a large impact into breeding decision-making process worldwide. Increasing current genetic variability is required in almond breeding programs to assure genetic gain and continuing breeding progress.

Cultivar Susceptibility and Environmental Parameters Affecting Symptom Expression of Red Leaf Blotch of Almond in Spain.

Red leaf blotch (RLB) of almond, caused by Polystigma amygdalinum, is an important foliar disease of this nut tree in the Mediterranean basin and Middle East regions. In recent years, the incidence of this disease has increased in Spain, corresponding to increases in the area of newly planted orchards and the use of susceptible cultivars. In 2009, an experimental orchard including 21 almond cultivars was planted at Les Borges Blanques, Lleida, in northeastern Spain. No fungicide treatments were applied during the 10-year experimental period (2009 to 2018) in order to allow natural disease development. Cultivar susceptibility to RLB was assessed each year, from 2011 to 2018, through visual observations of symptoms in naturally infected trees. The experimental results led us to classify the cultivars into five susceptibility groups. The most susceptible were Tarraco, Guara, Tuono, Marinada, Desmayo Largueta, and Soleta, whereas Mardía was the most tolerant. The annual incidence of disease was positively correlated with accumulated rainfall in spring, and especially in April, while it was negatively correlated with high spring and summer temperatures, especially in May. These findings could be used to improve disease management strategies by identifying the most susceptible cultivars and improving the timing of fungicide application.

Inoculum and Infection Dynamics of Polystigma amygdalinum in Almond Orchards in Spain.

Red leaf blotch (RLB) disease of almond, caused by Polystigma amygdalinum, is an important foliar disease in most production regions of the Mediterranean basin and the Middle East because severe infections may cause a premature defoliation of the tree. Some key aspects on the epidemiology of P. amygdalinum were studied in multiyear trials in two almond-growing regions in Spain, which included the seasonal development of perithecia and production and germination of ascospores along with the disease incubation and plant infectivity periods. Our results showed that primary inoculum was available in extended periods (January to August). Significant differences in ascospore amounts among regions, higher in the southern Andalusia and lower in the northern Catalonia, and years of study were detected. The factors geographical location, sampling period, and evaluation year were found significant on the development of P. amygdalinum perithecia. Variable ascospore germination rates were observed from April to July: >15% but rarely exceeding 30%. The RLB infectivity period in Catalonia extended from March to mid-June, whereas in Andalusia it was from March to May. The incubation period was mainly in a range of 5 to 10 weeks in Catalonia. The environmental conditions of October to January influence the available ascospore amounts in the next season. RLB infection occurs in spring to summer when mean temperatures are in the range 10 to 20°C. These results represent the first step in developing a prediction model of the disease that might serve as a tool for the control of RLB.