1. Introduction
Among the many pathogens capable of attacking
Prunus trees, brown rot caused by different species of
Monilinia is one of the most important economic factors limiting the production of stone fruit around the world [
1,
2]. In stone fruit,
Monilinia spp. are able to infect various plant organs, causing blossom blight, twig blight and brown rot in immature and mature fruits. The two main species responsible for attacks on flowers and twigs in
Prunus trees are
Monilinia laxa (Aderhold and Ruhland) Honey and
Monilinia fructicola (G. Winter). Apricot is the crop that is most susceptible to blossom and twig blight, followed in order by prune, sweet cherry, peach, sour cherry and plum trees [
3].
M. laxa can cause infections in apricot blossom, twigs and fruit. The first two are of the greatest concern, especially in organic production, causing losses of up to 90% in southern France [
4].
During winter,
M. laxa and
M. fructicola are preserved as mycelium in cankers on twigs infected the previous year and in mummified fruit which is hanging from branches or has fallen to the ground. For
M. fructicola, there may also be the formation of apothecia in mummified fruit, which then produce ascospores. Conidia and ascospores constitute the primary inoculum and can be transported by wind, rain and insects [
1]. When the conditions are humid, the flowers can be infected, with the mycelium progressing via the peduncle to reach the twigs and cause a canker there, which can lead to the apical section of the twig drying out [
2]. All parts of the flower can serve as the first infection site. There is evidence that fully open flowers are the most susceptible to infection [
3]. Luo et al. [
5] demonstrated that prune flowers have been shown to be the most sensitive to contamination by
M. fructicola when wide open (BBCH Stage 65) [
6]. In the orchard, the period of sensitivity of apricot flower buds to contamination by
M. laxa begins at the ‘sepals open’ stage (BBCH stage 57), increases until the ‘full flowering’ stage (BBCH stage 65) and ends at the ‘flowers fading’ stage (BBCH stage 67) [
6,
7,
8].
The presence of humid conditions at the time of flower susceptibility is crucial in paving the way for contamination by
Monilinia spp. Tamm et al. [
9] have shown in a growth chamber experiment the effects of petal wetness duration and temperature on the development of blossom blight caused by
M. laxa in cherry trees (
Prunus avium L.). They report that disease incidence increases with the duration of flower wetting. Luo et al. [
5] have shown similar results for
M. fructicola on plum flowers
(Prunus domestica L.). In an experiment with potted apricot trees installed in the orchard over short periods of 24 to 48 h, Tresson et al. [
7] confirmed that the duration of wetting was indeed an explanatory parameter for the incidence of
M. laxa on flowers, but that the amount of rain (in mm) was a more explanatory parameter than the duration of wetting. Therefore, they proposed a model making it possible to estimate, for each rainy episode, the incidence of
M. laxa on flowers according to rainfall and temperature for an orchard in which the aerial inoculum was between 50 and 200 conidia per m
3 of air [
8]. Rain could also play a role in the dispersal of conidia through a splashing effect. In a study conducted in California, Corbin and Ogawa [
10] showed that there was a greater dispersal of
M. laxa conidia in the air during rainy periods than in dry periods.
In some perennial crops, rain shelter systems have shown great effectiveness in reducing the incidence of many fungal diseases whose development requires a certain period of wetting in trees. For example, in Norway, Borve and Stensvand [
11] showed that the installation of transparent rain shelters on cherry trees during rainy periods from flowering to harvest made it possible to avoid fungicide protection while producing a relatively healthy harvest. In fact, covered trees presented on average only 3.4% of rotten cherries, mainly due to
Monilinia laxa and
Botrytis cinerea (Pers.). In contrast, unprotected trees received between three and six fungicide treatments from flowering to harvest and averaged 16.5% of rotten cherries [
11]. In France, the rain shelters offered by Filpack
® were evaluated from 2010 to 2015 on Braeburn and Gala apple varieties. In the absence of fungicide protection, the development of scab due to
Venturia inaequalis (Cooke) G. Winter was greatly reduced in sheltered trees compared to non-sheltered trees. Even in years with high scab pressure, such as 2012 and 2013, the symptoms on the leaves and fruits were very limited, with less than 3% of scabbed leaves and less than 1% of scabbed fruits in sheltered trees compared to 94% to 100% of scabbed leaves and 63% to 100% of scabbed fruits in non-sheltered trees [
12]. A study conducted on table grapes in southern China’s Yunnan Province showed the strong potential of rain shelters in reducing the development of major vine diseases. For two years, under the rain shelter, the severity of downy mildew on grapes due to
Plasmopara viticola (Berk. and Curtis) Berl. and de Toni was reduced by an average of 81% compared to the vines receiving fungicide protection, and by 94% compared to the control vines without fungicide protection. The severity of bunch rots due to
Colletotrichum gloeosporioides and
C. petrakii B. Sutton were reduced under rain shelters by 85% and 69%, respectively, compared to the vines receiving fungicide protection, and by 93% and 90% compared to the control vines without fungicide protection [
13].
In this study, we wanted to evaluate the advantages of installing rain shelters in orchards to protect apricot trees from moniliosis outbreaks on flowers and twigs. The first objective of this study was to evaluate the influence of transparent rain shelters on the modification of the microclimate at the level of the tree canopy and on the reduction in moniliosis damage to the twigs. The second objective was to test the effectiveness of protection strategies combining rain shelters and reduced fungicide protection against moniliosis in both conventional and organic production.
2. Materials and Methods
2.1. Experimental Orchards
The three experimental orchards were located at the INRAE-UERI experimental station in Saint-Marcel-lès-Valence (Drôme) in France’s Middle Rhone Valley. This is a continental area with Mediterranean summer influences.
All the apricot varieties used in the three orchards were grafted on Montclar® (4238) (Agri Obtentions, Guyancourt, France) rootstock. Trees were irrigated using localised micro-sprinkling under the tree foliage. This irrigation system avoids wetting the leaves and fruits during irrigation. There is no tree irrigation when the apricot trees bloom.
Transparent plastic rain shelters in high-density polyethylene (Anisolar, Filpack, Vitrolles, France), comprising two 1.5 m wide strips connected on a ridge cable above the tree row, were installed in the ‘Rain shelter’ mode of the three experimental trials, providing 3 m wide protection at a height of 3.5 m. The rain shelters were unrolled before the beginning of the sensitivity of flower buds to moniliosis contamination (
Table 1). They were folded up in July after fruit harvesting.
2.1.1. Experimental Orchard ‘Rain Shelter Protection Evaluation’
This orchard, planted in 1998, involved Bergarouge
® (2914) apricot trees, a cultivar with a high susceptibility to blossom and twig blight [
4]. The distance between trees was 3.5 m, with a 5 m gap between rows. In October 2013, the trees, initially pruned in ‘gobelet’ style, were pruned to form a rectangular parallelepiped 3 m high and 2 m wide to ensure trees could be protected from rain under the shelters. Control trees were pruned in the same fashion. Treatments (‘Rain shelter protection’ and ‘Control without protection’) were replicated twice with a block design. The experimental system comprised 12 trees under rain shelters and 12 trees without rain shelters.
The rain shelters were unrolled on 24 February 2014 and 3 March 2015. No fungicides were applied to control blossom and twig blight in 2014 and 2015.
2.1.2. Experimental Orchard ‘Rain Shelter and Fungicide Use Reduction’
This orchard was established in January 2015. It has five varieties, including Bergeron (660), Bergeval
® (3950) (CEP Innovation, Lyon, France), Shamade (3902) and Anegat (4481), which are considered susceptible to moniliosis on flowers and twigs, as well as Frisson (2821), which is considered very susceptible to moniliosis [
4]. Each elementary plot consists of a row of eight trees of the same variety, with a planting distance of 2.5 m between trees and 4 m between rows. The trees were trained in trellised ‘palmettes’ so as to form a fruit wall about 1 m wide. The ‘protection system’ study factor is made up of two modes, one without rain shelters with classic fungicide protection (called ‘Fungicide reference’), and one with rain covers combined with reduced fungicide protection (called ‘Rain shelter’). Treatments were replicated twice with a block design. The trial involved 80 trees under rain shelters and 80 trees without rain shelters.
The rain shelters were unrolled on 11 February 2016, 21 February 2017 and 5 March 2018. During the period when flower buds are sensitive to contamination, preventive fungicide protection was used, taking into account rainy episodes announced in weather forecasts. Fungicide treatments were renewed when new flowers opened after the previous application. The reduction in fungicide protection for the ‘Rain shelter’ mode consisted of removing the last fungicide application positioned at the end of the period of flower sensitivity to contamination by moniliosis (
Table 2). No fungicide protection against moniliosis on flowers was conducted in 2015 because there were no flowers on the trees during the first year of growth in the orchard.
2.1.3. Experimental Orchard ‘Organic Orchard without Copper’
This experimental system was established in January 2020 and is managed according to the French rules for organic production. It includes two varieties, Vertige (3845), which is considered susceptible to moniliosis on flowers and twigs, and Tom Cot
® (2669) (Cot International, Bouillargue, France), which is considered fairly insensitive [
4]. The elementary plots consist of five rows of six trees, i.e., thirty trees of the same variety. Planting distances between trees are 2.5 m, with 4 m between rows. The ‘protection system’ study factor comprised three modes as follows: (i) an ‘Organic farming reference’ mode in which the trees are grown ‘gobelet’ style and for which a classic organic farming fungicide protection programme against moniliosis is applied, (ii) a ‘Control’ mode in which the trees are grown ‘gobelet’ style without fungicide protection against moniliosis provided, and (iii) ‘Rain shelter’ mode in which the trees are trained in a ‘palmette’ 1 m wide under rain shelters and for which no fungicide protection against moniliosis is provided. Both varieties were included in all three modes, with treatments replicated twice with a block design. The experiment involved 360 trees, including 120 trees under rain shelters.
Rain shelters were unrolled on 18 February 2021 and 23 February 2022. During the period when flower buds are sensitive to contamination, fungicide protection in the ‘Organic farming reference’ mode was conducted (i) either preventively with copper sulphate taking into account rainy episodes announced in weather forecasts, or (ii) at the end of the rainy period with calcium polysulphide (
Table 3).
2.2. Twig Blight Assessment
For the ‘Rain shelter protection evaluation’ and ‘Rain shelter and fungicide use reduction’ trials, the damage assessment of Monilinia spp. on twigs was carried out by counting the number of twigs dried out by Monilinia spp. per tree.
For the ‘Organic orchard without copper’ trial, the twig blight assessment consisted of an observer visually evaluating Monilinia spp. infection in the tree. This evaluation was carried out 30 days after full blossom. For each tree, the observer visually estimated the ratio between the total length of flower-bearing twigs dried out by Monilinia spp. and the total length of flower-bearing twigs. This meant that the scale used was from 0 to 100% (total infection).
In all trials, once scoring was completed, all moniliated twigs were cut and removed from the orchard. This prophylactic action makes it possible to restore each of the different modes to a primary inoculum level of the same order of magnitude for the year n + 1.
2.3. Identification of the Monilinia Species Responsible for Moniliosis on Flowers and Twigs
The identification of
Monilinia species began with the collection of orchard samples of flowers and moniliated twigs. Moniliated flowers were placed directly in a Petri dish containing a PDA medium (potato dextrose agar, Conda Laboratory, Spain; 9.75 g of PDA medium for 250 mL of deionised water). To take isolations from the twigs, a portion of the twig was taken from under the bark at the boundary between the healthy and necrotic zones and placed in a Petri dish. After 10 days of growth of the mycelium, identification was made according to the morphological characters of the mycelium defined by the Lane’s synoptic key [
14].
2.4. Climatic Conditions and Canopy Microclimate
Climatic data were recorded by an Enerco 516i meteorological station (Cimel Electronique, Paris, France) from INRAE’s national Agroclim network, located on grassland between 300 and 900 m from the experimental orchards.
To assess the effect of rain shelters on microclimatic conditions during the ‘Rain shelter protection evaluation’ and ‘Rain shelter and fungicide use reduction’ trials, air temperature, air relative humidity and leaf wetness were recorded within tree canopies. Air temperature and relative humidity were measured using iButton
® temperature/humidity loggers (THB—DS1923, Dallas Semiconductor, Dallas, TX, USA). All loggers were protected in well-ventilated white shelters of 950 cm
3 to prevent them from direct sunlight and provide airflow. Leaf wetness duration (LWD) was measured using home-made artificial leaves built from a rectangular electrical insulator comprising two interdigitated electrodes. The LWD estimate is based on determining the electrical resistance on the surface of the sensor [
15]. The estimation of the LWD was conducted as follows: First and prior to the experiment, the resistivity (in ohms) of the sensor was estimated in controlled conditions for dry and wet conditions. A threshold value was then obtained that enables us to indicate whether liquid water is present (LWD is set to 1) or not (LWD is set to zero) at the sensor surface. In practice and in the experiment, the LWD was estimated by converting the electrical resistance measured by the sensors in 0 and 1 according to the threshold value. This value was multiplied by acquisition time (for instance, 10 min) to obtain the LWD. The procedure was applied to the entire dataset to obtain the LWD. The values were summed to obtain the monthly values.
For the ‘Rain shelter protection evaluation’ trial, three trees per block were equipped with a LWD sensor, and two trees per block were equipped with a THB sensor. Each LWD sensor was located at a height of 1.8 m and 0.6 m from the tree trunk following the row axis and on the north side of the tree. THB sensors were attached to the north side of the trunk at a height of 1.8 m. Data were collected from February to October every 10 min during 2014 and 2015. For the ‘Rain shelter and fungicide use reduction’ trial, six LWD and two THB sensors per mode were mounted within tree canopies. For each mode, sensors were located at a height of 1.8 m and distributed around two trees (three sensors per tree for LWD and one THB per tree) following the north-western, north-eastern and southern directions relative to the trunk. Data were collected from February to September every 10 min in each year.
2.5. Data Analysis
The number of twigs per tree dried out by
Monilinia spp. and the proportion of necrotic twigs showing symptoms of
Monilinia spp. were subjected to an analysis of variance (ANOVA) using Statgraphics Plus 5.1 software (Manugistics, Rockville, MD, USA). The level of significance was set at 5% for all the statistical tests. The normal distribution of ANOVA residuals was checked using the Shapiro–Wilk test [
16], and the independence of ANOVA residuals and the intra-treatment variance equality (homoscedasticity hypothesis) were visually checked using the residuals/predicted values graph. When residual standard deviation increased with the increment of predicted values, data were log-transformed before ANOVA [
17]. Mean comparisons were conducted using the Newman–Keuls test.
For the ‘Rain shelter protection evaluation’ trial (‘Organic orchard without copper’), each year, the number of moniliated twigs per tree (the percentage of necrotic twigs) was subjected to a two-factor variance analysis (treatment factor and block factor).
For the ‘Rain shelter and fungicide use reduction’ trial, each year, the number of moniliated twigs per tree was subjected to a two-factor variance analysis (treatment factor and variety factor). In the absence of significant treatment x variety interaction, the results are presented for the average of the five varieties.
The entire leaf wetness duration dataset was separated into leaf wetness duration due to rain and dew by considering rain events and clear night sky conditions, respectively, identified from the meteorological data.
4. Discussion
In the first trial, conducted without fungicide protection, the rain protection demonstrated a partial efficacy of 25% to 62% in limiting moniliosis damage on twigs. This relatively low efficacy could be partly due to the fact that the trees were initially pruned in a broad form (‘gobelets’) before being pruned to form a hedge only 2 m wide. Protection with shelters that were only 3 m wide was perhaps not sufficient. In the two subsequent trials, the trees were pruned in a ‘palmette’ pattern to form a fruit hedge just 1 m wide, over which protection with 3 m wide rain shelters was installed. In 2017, the two modes tested received exactly the same fungicide protection, and a fairly substantial reduction in the number of moniliated twigs of 95% was observed in the ‘Rain shelter’ mode. In 2021 and 2022, the strong efficacy of rain covers was also observed, with an 89% to 97% reduction in moniliosis damage. This significant reduction in the number of affected twigs under rain shelters is certainly linked, in part, to the good protection against rain provided by cultivation in a narrow fruit hedge.
In 2018, reduced fungicide protection was used in the ‘Rain shelter’ mode compared to the ‘Fungicide reference’ mode, but it still demonstrated a 65% reduction in the number of moniliated twigs. It can be hypothesised that partial-effect cultural methods are more effective when combined with fungicide protection. For example, Didelot et al. [
18] showed that varietal mixtures of apple trees combining a variety resistant to scab (due to
V. inaequalis) with a susceptible variety was more effective when combined with reduced fungicide protection. In their study, the reduction in the incidence of leaf scab in the susceptible variety grown in a mixture compared to the susceptible variety as a single crop was 7% to 20% in the absence of fungicide protection and 41% to 75% with reduced fungicide protection.
As expected, the rain shelters significantly reduced the leaf wetness duration (LWD) measured within the foliage compared to the unsheltered trees (a reduction of 43% to 67%). The rain shelters reduced wetting times both during and after a rain event and dew deposition times after a clear night. Overall, the rain shelters led to a greater reduction in dewy conditions than in rainy ones (
Table 4 and
Table 6). This is consistent with the LWD observed in the tree canopies [
19] and the physics of water deposition by rain and dew [
20]. Indeed, a rain shelter acts as an effective physical barrier against water droplets from rain but also prevents leaves from night-time cooling by limiting longwave energy losses during clear sky conditions. However, rain shelters also limit the evaporation process by decreasing net radiation [
12] and then increasing evaporation time [
20]. The sensors were located at different locations within the canopies and it has been shown that the tree canopies intercept part of the rain droplets, and then lead to a reduction in the LWD within the canopies [
21]. This means part of the observed reduction in the LWD may also result from a canopy effect.
Very few differences were observed in the daily averaged air temperature (up to 6%) and the daily averaged air relative humidity (up to 1%). So, the overall decrease in ‘damage’ in trials one and two may be mainly due to the reduced LWD as a result of the use of rain shelters and not by changes in the air properties. The rain protection, by limiting the direct impact of raindrops on the tree canopies, could also limit the dispersion of conidia present on cankers and mummified fruits.
In the third experimental approach, managed according to organic principles, the levels of moniliosis damage on the twigs of the system with rain protection and no fungicide protection are lower or equivalent to the organic reference system using fungicide protection. These results make it possible to envisage organic apricot production without the use of cupric fungicides [
22].
Rain protection has also been shown to be highly effective in reducing the development of rust epidemics in apricot trees by limiting primary contamination in May and June [
23,
24]. However, it is difficult to assess the direct effectiveness of rain shelters on moniliosis on fruits at harvest. Indeed, because apricot fruits are not very sensitive to moniliosis, the rotten fruits found at harvest are often those that have suffered damage (earwig bites, hail impact, etc.). However, the installation of rain shelters requires numerous posts and anchor cables which allows earwigs to pass towards the canopy of trees and so limits the effectiveness of physical barriers such as the installation of glue bands on the trunk. On the other hand, rain shelters greatly reduce fruit damage due to hail.
Providing rain protection could be an interesting solution for producing organic apricots without resorting to copper-based fungicides, the environmental impact of which is not insignificant [
25,
26].