Host range of Phytophthora spp. from berry crops in Huelva, Spain.

Crop declines have been observed in raspberry and blueberry farms in the southwest region of Spain, which is the most important berry-producing area in the country. This study aimed to identify and characterize the pathogens associated with these diseases using molecular and morphological methods. Additionally, pathogenicity tests were performed on different raspberry, blueberry, and strawberry cultivars to determine possible susceptible hosts in the area. An isolate of P. cactorum was obtained from a symptomatic strawberry plant, an isolate of P. cinnamomi was obtained from a symptomatic blueberry plant, and isolates identified as P. rosacearum, P. rubi and a previously unknow speciesrecently named as P. sp. balkanensis were recovered from symptomatic raspberry plants. Results from the pathogenicity tests reported, for the first time, P. rubi causing root rot and wilting complex (RRWC) in Spanish raspberry crops. Additionally, P. cinnamomi was found to affect highbush blueberry production in Spain. Thus, this study provides valuable insights into the identification and characterization of Phytophthora spp. associated with the decline of blueberry and raspberry crops in Huelva. It also provides essential recommendations regarding the potential risks associated with the use of other types of berries as rotational crops and emphasizes the necessity for effective management strategies to mitigate crop losses. This is particularly critical given the limited soil disinfection alternatives available in Spain.

First report of root and crown rot caused by Fusarium oxysporum Schltdl. on Prunus cerasifera L. in Spain.

In 2021, Spain was the largest producer of cherries in Europe with a production of 125810 tons (FAOSTAT, 2021). In May 2022, in several production fields in Huelva (Spain), wilting was noted in 4-year-old cherry trees cv. Crystal Champaign grafted on rootstock cv. Adara (Prunus cerasifera L.). Gumming and wilting affected approx. 1% of the production area, leading to the eventual collapse and death of most affected trees within 2-3 years. Discoloration of the vascular system of the crown and roots was also noted. Symptomatic crown and root pieces (0.5 cm) were subjected to surface sterilization: immersed in 1% NaClO for 2 min, rinsed in sterile distilled water, and air-dried in a laminar flow cabinet. Then, plant tissues were placed on potato dextrose agar (PDA) amended with streptomycin and incubated in a lab bench at room temperature for a week. Cottony and pink colonies were observed growing from the tissues. Two single strains (F175 and F176) were obtained from each tree by excising single spores (Gordon and Okamoto 1991). Isolates produced sparse aerial mycelia with white to pinkish-orange pigmentation on Spezieller Nährstoffarmer Agar (SNA). Both isolates produced microconidia in false heads on short monophialides. Microconidia were hyaline and measured in the range of 5.0-17.5 × 2.5-3.8 µm for both isolates (n = 50). Macroconidia were less abundant, falciform, and hyaline. Morphological characteristics were consistent with identification as Fusarium spp. (Leslie and Summerell 2006). A portion of the translation elongation factor-1 alpha (EF-1α) gene was sequenced using EF1/2 primers (O'Donnell et al. 1998) (GenBank Accession Nos. OR733348 and OR733349). Based on a comparison of 619 base pairs (bp), both isolates exhibited different sequences, with a 99.5% similarity (616/619 bp). A comparison with previously described isolates revealed a 100% match with published F. oxysporum sequences in the GenBank database (KT323846 and MZ404079, respectively). Isolates were used to conduct pathogenicity tests on 1-year-old plants cv. Adara growing in 512 cm3 pots. Using a scalpel, a 6-7 mm-length wound (2-3 mm deep) was made 5 cm above the soil line in all plants. For each isolate, 5 plants were inoculated by placing a 5 mm plug containing 10-day-old mycelia grown in AMAP medium (Borrero et al. 2009) at the incision point. Non-colonized AMAP plugs were used to inoculate 5 control plants. The inoculated sites were sealed with parafilm. Plants were randomly placed in a growth chamber with a temperature of 28/24ºC and a 12-hour photoperiod. A reddish-brown vascular stem discoloration was noticed in all the inoculated plants 73 days after inoculation. On average, the length of the necrotic area was 12.73 cm for F175, 20.12 cm for F176, and 4.59 cm for the control plants. Fusarium oxysporum was successfully re-isolated from all the inoculated plants. Recovered isolates were confirmed to be the same as the inoculated ones by sequencing the EF-1α gene. A one-way ANOVA indicates that plants cv. Adara were susceptible to both F. oxysporum isolates (P < 0.05). This is particularly noteworthy as cherries are predominantly planted on rootstocks and cv. Adara is a widely used rootstock in Spain. While F. oxysporum has been reported as the cause of root and crown rot in sweet cherry (P. avium L.) in British Columbia (Úrbez-Torres et al. 2016), this is the first report of F. oxysporum causing root and crown rot in cherry rootstocks (P. cerasifera L.) in Spain.

Potential Inoculum Sources and Incidence of Strawberry Soilborne Pathogens in Spain.

The decline and death of strawberry plants in Spanish fruit production fields have mainly been attributed to the soilborne pathogens Macrophomina phaseolina, Phytophthora cactorum, and Fusarium spp. Inoculum sources of M. phaseolina and P. cactorum, and the incidence all three genera, were investigated in nurseries and fruit production fields over three consecutive seasons. M. phaseolina inoculum sources consisted of fumigated preplant fruit production soils (50%) and fumigated nursery soils (47%), although the pathogen could not be detected in nursery mother and runner plants. P. cactorum inoculum sources included nursery (20%) and preplant fruit production (17%) fumigated soils, and nursery runner plants (up to 15%). In fruit production plants, the average incidence of M. phaseolina and P. cactorum were 4.2 and 3.7%, respectively. Fusarium spp. inoculum sources could not be accessed extensively due to the lack of effective quantitative real-time PCR assays. Limited testing of nursery plants showed that Fusarium oxysporum f. sp. fragariae (Fof) was absent. In field production plants and soil, F. solani was the main pathogenic Fusarium spp., with Fof only identified once in a fruit production plant. Ineffectively fumigated soils in nurseries and production fields, along with infected runner plants, can be inoculum sources of soilborne strawberry pathogens in Spain.