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BACKGROUND Terrestrial mollusks are one of most important agricultural pests worldwide. Natural phytochemicals have an extended history as a source of pesticides. This study was planned to isolate molluscicidal active compounds from the stems of Adenium obesum. RESULTS: The benzene‐soluble fraction of the hydroethanolic extract displayed the most potent molluscicidal activity against Monacha obstructa among different solvent fractions with a median lethal dose (LD50) of 4.91 µg g−1 body weight (bw). The bioactivity‐guided chemical exploration of the benzene‐soluble fraction led to the isolation of two known cardiac glycosides, cerberin and neriifolin which showed significant molluscicidal activity with LD50 values of 5.39 and 4.3 µg g−1 bw, respectively. CONCLUSION: Isolation of the cardiac glycoside neriifolin from A. obesum and the molluscicidal activity of cerberin and neriifolin against terrestrial snails are reported for the first time. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Plants have evolved different defense mechanisms to combat pathogen attacks. In this study, detached leaf assays were conducted to estimate the influence of leaf position (leaf age) on the development of chocolate spot disease, caused by Botrytis fabae, in the lower, middle and upper leaves of five faba bean (FB) cultivars with different levels of resistance/susceptibility. Two components of resistance in terms of lesion diameter and spores per lesion were used to evaluate the disease intensity. To evaluate plant defense response, oxidative burst and phenol-oxidizing enzymes were assayed. The results indicated that regardless of the resistance level of the FB cultivar, lower (older) leaves were more severely infected by the chocolate spot pathogen than upper (younger) ones. A comparison of the defense-response behavior of the FB leaves revealed that the chocolate spot pathogen induced lipid peroxidation and the production of reactive oxygen species, peroxidase, and polyphenol oxidase in leaf tissue during the FB-B. fabae interaction. The production of these defense compounds in leaves was not static but governed in time and extent by physiological maturity. Younger leaves exhibited significantly higher oxidizing enzyme activity and lower oxidative stress than older ones. These variations in the levels of defense compounds could explain the differences in leaf resistance. The extreme differences in disease development recorded in upper and lower leaves of all FB cultivars suggest that assessing resistance using leaves from the middle positions would be the most efficient and reliable for evaluating the resistance/susceptibility of FB plants to B. fabae. [ABSTRACT FROM AUTHOR]

Recently, Saudi growers have expanded their production of organic, soilless-grown strawberries (Fragaria × ananassa Duch.), but their production shows many difficulties associated with disease susceptibility. In October 2021, 45% of strawberry plants cv. "Festival" organically cultivated in Dammam city, Saudi Arabia (26°31'34.5"N 50°00'51.0"E) showed wilting symptoms. Typical symptoms were yellowing, rapid wilting, death of older leaves, stunting, and decreased roots. Vascular bundle necrosis and crown and root rot were also observed; plants eventually collapsed and died. Twenty symptomatic strawberry plants were sampled to isolate the pathogen. Pieces (4 × 4 mm) of the symptomatic tissues from crowns and roots were sanitized with 1% NaOCl (90 s), submerged in 70% alcohol (20 s), rinsed with sterile water (2x 30 s), placed on potato dextrose agar (PDA; Scharlau Chemie, Spain) and incubated at 25°C for 6 days. Next, we prepared single-spore cultures on PDA and synthetic nutrient-poor agar (SNA). On PDA media, pure cultures produced abundant aerial mycelium, with light pink or purple pigmentation in the medium after incubation at 25°C for 7 days. On SNA media, aerial microconidia were abundant cylindrical to ellipsoid hyaline with zero to one septate (3.8 - 5.9 × 1.3 - 2.5 μm, n = 50). Macroconidia were few, hyaline and falcate, with slightly curved apexes and 2 to 4 septate (18.9 - 27.5 × 3.3 - 4.6 μm, n = 50). Chlamydospores were roundish and terminal or intercalary. As Leslie and Summerell (2006) described, such morphological characteristics are typical of F. oxysporum. The isolates' identities were established by extracting DNA using the DNeasy Plant Mini kit (QIAGEN, Hilden, Germany). This was followed by amplification and sequencing of the internal transcribed spacer (ITS) (White et al., 1990), elongation factor 1-α (TEF1-α) (O'Donnell et al., 1998), and the ribosomal RNA intergenic spacer (IGS) (Canizares et al., 2015). The ITS, TEF1-α, and IGS sequences of an isolate Fof-10 were submitted to GenBank (PP564462, PP703242, and PP784894, respectively). BLAST analysis confirmed 99.71 and 100% identities to the ITS, TEF-1α, and IGS sequences of F. oxysporum (KU931552.1, OR640020.1, and FJ985519.1), respectively. All isolates tested were confirmed at the forma specialis fragariae, level using the specific primers FofraF/FofraR (Suga et al. 2013). The ∼239 bp amplicon was sequenced and submitted to GenBank (PP703243). Two-month-old healthy strawberry plants of cultivars "Festival," "Marquis," and "Monterey" were inoculated by dipping the roots in the spore suspension (107 conidia ml-1) for 15 min (Henry et al. 2017). There were five replicates for each cultivar. Plants dipped in water were used as a control treatment. The plants were transplanted in sterilized soil and placed in a greenhouse at 30/26°C (day/night). Within 4 to 6 weeks, inoculated plants showed severe wilting and discoloration of the internal crown tissue, while control plants were symptomless. The pathogen was re-isolated from the discolored vascular tissue onto PDA and identified morphologically and molecularly as the original one, thus fulfilling Koch's postulates. The test was repeated twice. This report confirms F. oxysporum f. sp. fragariae as a causal agent of Fusarium wilt of strawberries in Saudi Arabia. This pathogen was previously reported to cause the Fusarium wilt of strawberries in California (Dilla-Ermita et al., 2023). This disease has been observed in several hydroponic strawberry greenhouses in Saudi Arabia, with incidence ranging from 25% to 45% across multiple locations. Given this, proper strategies are needed to manage this disease and to be compatible with organic farming.

Citrus greening disease was first reported in Saudi Arabia during the 1970s when characteristic foliar and fruit symptoms were observed in commercial citrus groves. However, ' Candidatus Liberibacter asiaticus' (CLas) was not detected in symptomatic trees until 1981 to 1984 when CLas-like cells were observed by transmission electron microscopy in leaves collected from symptomatic citrus groves in southwestern Saudi Arabia. Despite the anticipated establishment of the CLas-Asian citrus psyllid (ACP) ( Diaphorina citri Kuwayama) pathosystem, CLas presence has not been verified in suspect trees nor have ACP infestations been documented. Given the recent expansion of citrus production in Saudi Arabia, a systematic country-wide survey was carried out to determine the potential CLas distribution in the 13 citrus-growing regions of the country. Citrus trees were surveyed for the presence of CLas-psyllid vector(s) and characteristic disease symptoms in commercial and urban citrus trees. Adult psyllids collected from infested citrus trees were identified as ACP based on morphological characteristics. Real-time quantitative PCR amplification (qPCR) of the CLas β-subunit of the ribonucleotide reductase (RNR) gene from citrus leaf and fruit samples and/or ACP adults revealed that trees were positive for CLas detection in 10 of the 13 survey regions; however, CLas was undetectable in ACP adults. Phylogenetic and single nucleotide polymorphism (SNP) analyses of a PCR-amplified, cloned fragment of the CLas 16S rRNA gene (∼1.1 kbp) indicated Saudi Arabian isolates were most closely related to Florida, U.S.A., isolates. Analysis of climate variables indicated that the distribution of the ACP-CLas pathosystem observed in Saudi Arabia was consistent with published predictions of terrains most likely to support establishment., Competing Interests: The author(s) declare no conflict of interest.

Cucumber (Cucumis sativus L.) is an important vegetable crop in Saudi Arabia. During May 2018, 45 - 60% of 5-month-old cucumber plants showed symptoms of a previously unknown wilt in commercial greenhouses around Al Kharj area of Riyadh region. Symptoms consisted of crown and root rot, wilting and stem disintegration, along with yellowish brown to brown external discoloration extended throughout the affected tissues. As the disease progressed, a pinkish-orange mycelial growth was often observed at the basis of affected stems while vessels were discolored. Subsequently, the affected plants were collapsed and died. Crown, stem, and root fragments (4 × 4 mm) were cut from symptomatic tissues, surface sterilized in 2.5% NaOCl, cultured on potato dextrose agar (PDA) with 25 mg/liter of streptomycin sulfate, and incubated at 26°C in darkness for 6 days. Single-spored cultures produced white mycelium with pink, white, or purple pigmentation in the center. The mycelium produced sporodochia. Macroconidia were mainly slightly curved with three to five septa. Microconidia were single-celled oval and produced on short lateral phialides. Chlamydospores were single or in short chains. Morphologically, the isolated fungus was characterized as Fusarium oxysporum (Leslie and Summerell 2006). To further confirm the fungus identification, DNA was extracted from a single-spored culture. Three different fungal nuclear regions of internal transcribed spacer (ITS), elongation factor 1-α, (TEF1-α) and the second largest subunit of DNA-directed RNA polymerase II (rpb2) with the following primers: ITS4 and ITS5 (White et al. 2017), EF-1 and EF-2 (O'Donnell et al. 2008), and fRPB2-5F and fRPB2-7cR (Liu et al. 1999), respectively. The ITS, TEF1-α, and rpb2 sequences of the isolate FCKSU17 were submitted to GenBank (MT232918, MW471131, and MW449833 respectively). Phylogenetic analysis based on the alignment of the ITS, TEF1-α, and rpb2 sequences using MEGA7 placed this strain in the F. oxysporum clade. To confirm the forma specialis radicis-cucumerinum, amplification with the specific primers ForcF1/ForcR2 was conducted (Lievens et al. 2007). The amplified fragment (∼ 250-bp) was sent for sequencing, and the sequence was submitted to GenBank (MW471132). BLASTn analysis of the sequences showed 100% identity with F. oxysporum radicis-cucumerinum (KP746408). To fulfill Koch's postulates, pathogenicity test was conducted on 7-day-old plants of cucumber cultivar Beit Alpha grown into pots filled with soil mix (2:1 sandy loam-peat moss, vol/vol). The plants were inoculated through drenching with 100 ml of conidial suspension in sterile distilled water (106 spores/ml) per pot. Control plants were treated with sterile distilled water. Each treatment included 10 replicates (pots), with two plants per pot. The pathogenicity test was repeated once. Cucumber plants inoculated with the fungus showed early wilting symptoms within the first 2 weeks post inoculation. At the 6th week post inoculation, 90 to 100% of the inoculated plants developed typical symptoms. No symptoms were observed on the control plants. The pathogen was successfully re-isolated from the inoculated wilted plants and identified morphologically. To our knowledge, this is the first report of F. oxysporum f.sp. radicis-cucumerinum on cucumber in Saudi Arabia. It is recommended that preventive management should be considered as this disease may cause significant economic losses on cucumbers in Saudi Arabia.

Bayoud disease, caused by Fusarium oxysporum f. sp. albedinis, is a very serious and destructive disease to date palm. Screening of date palm germplasm for resistance to bayoud disease is a crucial step to avoid or alleviate the disease consequences. Fortunately, it was discovered that there are two mitochondrial plasmid-like DNA molecules associated with susceptibility or resistance to bayoud disease. In this chapter, we present a fast, simple, and reliable technique to screen date palm germplasm for the presence of these mitochondrial molecular markers associated with susceptibility or resistance to bayoud.