Assessment of genetic diversity and phylogenetic relationship of local coffee populations in southwestern Saudi Arabia using DNA barcoding.

The genetic diversity of local coffee populations is crucial to breed new varieties better adapted to the increasingly stressful environment due to climate change and evolving consumer preferences. Unfortunately, local coffee germplasm conservation and genetic assessment have not received much attention. Molecular tools offer substantial benefits in identifying and selecting new cultivars or clones suitable for sustainable commercial utilization. New annotation methods, such as chloroplast barcoding, are necessary to produce accurate and high-quality phylogenetic analyses. This study used DNA barcoding techniques to examine the genetic relationships among fifty-six accessions collected from the southwestern part of Saudi Arabia. PCR amplification and sequence characterization were used to investigate the effectiveness of four barcoding loci: atpB-rbcl, trnL-trnF, trnT-trnL, and trnL. The maximum nucleotide sites, nucleotide diversity, and an average number of nucleotide differences were recorded for atpB-rbcl, while trnT-trnL had the highest variable polymorphic sites, segregating sites, and haploid diversity. Among the four barcode loci, trnT-trnL recorded the highest singleton variable sites, while trnL recorded the highest parsimony information sites. Furthermore, the phylogenetic analysis clustered the Coffea arabica genotypes into four different groups, with three genotypes (KSA31, KSA38, and KSA46) found to be the most divergent genotypes standing alone in the cluster and remained apart during the analysis. The study demonstrates the presence of considerable diversity among coffee populations in Saudi Arabia. Furthermore, it also shows that DNA barcoding is an effective technique for identifying local coffee genotypes, with potential applications in coffee conservation and breeding efforts.

Diversity of arbuscular mycorrhizal fungi associated with acacia trees in southwestern Saudi Arabia.

Acacia species produce extensive, deep root systems with a capacity to develop mycorrhizal symbioses that facilitate plant nutrition via enhanced soil nutrient absorption. This study aimed to evaluate the mycorrhizal status and the diversity of arbuscular mycorrhizal fungi (AMF) associated with acacia trees in southwestern Saudi Arabia. The mycorrhizal status varied greatly between species. The highest values of AMF root colonization and spore density were observed in the roots and in the rhizospheric soil of Acacia negrii. DNA was extracted from plant roots and the AMF large subunit ribosomal DNA (LSU rDNA) was amplified by a nested polymerase chain reaction. A total of 274 LSU rDNA cloned fragments from roots of the three acacia trees were sequenced. Phylogenetic analysis revealed a high AMF diversity, especially in Acacia tortilis. On the basis of LSU rDNA sequences, AMF was grouped into five genera: Glomus, Claroideoglomus, Acaulospora, Gigaspora, and Scutellospora. The genus Glomus fungi were the dominant colonizers of all three acacia species, while the genus Scutellospora fungi were found only in A. tortilis roots. The high AMF-acacia diversity suggests that AMF plays an important role in the sustainability of acacia species in the arid environment.

Ethnobotanical Study of Indigenous Medicinal Plants of Jazan Region, Saudi Arabia.

For a long time, the people of Saudi Arabia have been using medicinal plants (MPs) as conventional medicine to heal diverse human and livestock diseases. The present work is the first study on ethnobotanical uses of 124 MPs species used by the local tribal communities of Jazan province in the Southwest of Saudi Arabia. Ethnobotanical data were collected by interviewing 174 local informants using semistructured interviews. Informants of different ages, from several settlements belonging to several tribal communities, were interviewed. It is worth noticing that the age of informants and their knowledge of MPs were positively correlated, whereas the educational level and MP knowledge of participants were negatively correlated. To find out if there was agreement in the use of certain plants in the treatment of given ailments, we used Informant Consensus Factor (ICF). To determine the most frequently used plant species for treating a particular ailment category by local people we used the fidelity level (FL%). The Relative Frequency of Citation (RFC) was used to indicate the local importance of a species and the relative importance (RI) level was used to check the therapeutic potentials of the cited plants. A total of 124 MPs belonging to 103 genera and 48 families were collected and identified. The majority of these plants were shrubs (45%), perennial herbs (21%), annual herbs (19%), or trees (18%). The Asteraceae (10.48%), Fabaceae (7.25%), and Apocynaceae (7.25%) families were the most represented. Leaves, fruits, and whole plant (24%, 18%, and 16%, respectively) were the most used plant parts in formulating traditional medicines. Ziziphus spina-christi and Calotropis procera with the highest RI level (2.0) were found to have the highest range of therapeutic uses. They were followed by Datura stramonium (1.86), Withania somnifera, and Aloe vera (1.81). The ICF ranged from 0.02 to 0.42 covering 12 disease categories with a prevalence of disease categories related to skin and hair problems (ICF=0.42) having 75 species cited, while 73 species were cited for gastrointestinal tract (GIT) disorders (ICF = 0.40). Senna alexandrina (67%), Tribulus terrestris (64%), Pulicaria undulata (60%), Leptadenia pyrotechnica (55%), and Rumex nervosus (55%) had the highest FL which indicates their good healing potential against specific diseases. The high-FL species are the most promising candidate plants for in-depth pharmacological screening and merit further consideration. Accordingly, Jazan flora has good ethnobotanical potential. Unfortunately, many MP species are endangered by drought, overgrazing, and overexploitation. Some protection measures should be undertaken to prevent these species from becoming extinct. Natural reserves and wild nurseries are typical settings to retain medically important plants in their natural habitats, while botanic gardens and seed banks are important paradigms for ex situ conservation.

Molecular identification of biological contaminants in different drinking water resources of the Jazan region, Saudi Arabia.

Drinking water quality plays a remarkable role in human infections and diseases. This study used polymerase chain reaction (PCR) techniques to detect bacterial pathogens. In addition, a physicochemical analysis was performed on drinking water samples from several sources. A total of 123 drinking water samples were collected from different areas in the Jazan region in Saudi Arabia: ground water (40 samples), bottled water (15 samples), tap water (52 samples), and water purification shops (16 samples). To isolate the bacterial pathogens, the water samples were spread on Nutrient and MacConkey agar media, and the grown pathogens were then identified by the 16S ribosomal RNA technique. In 87 (70.7%) of the 123 drinking water samples, there was no pathogen growth on the two-culture medium. However, 36 (29.3%) of the samples were found to be contaminated with bacteria. The physicochemical analysis indicated that the water samples were within the Saudi drinking water standards. The bacteria were resistant to Cefotaxime, Cefotaxime/Clavulanic acid, Erythromycin, Penicillin G, Rifampin and Sulfamethoxazole-Trimethoprim, respectively. The findings suggest that in Jazan, bottled water is a safer source of potable water than tap water. The contamination in the water may be occurring at the reservoirs rather than the water sources.

Arbuscular mycorrhizal fungi in soil, roots and rhizosphere of Medicago truncatula: diversity and heterogeneity under semi-arid conditions.

Mycorrhizal symbioses are considered indicators of ecosystem biodiversity. However, their diversity and relevance in arid and semi-arid ecosystems are poorly understood. This study addressed this subject, the main objective being to evaluate arbuscular mycorrhizal fungi (AMF) diversity and heterogeneity in a semi-arid region. Samples of bulk and rhizosphere soil and fine roots of Medicago truncatula were collected at four different sites with the same aridity index (6.1), in Bou-Hedma National Park, Tunisia, a semi-arid ecosystem. AMF taxa were assessed by 454- pyrosequencing and identified by BLAST matching of operational taxonomic units (OTUs) against the MaarjAM database, targeting AMF SSU rRNA gene diversity. Roots were the hotspots of AMF diversity (107 OTUs out of a total of 138). Of the 138 OTUs, 113 found correspondence in the MaarjAM database, with 32 AMF virtual taxa (VTX),19 Site-exclusive (SE) and 13 common to at least two sites (Non-site exclusive, NSE); the remaining 25 OTUs grouped in 16 putative new AMF taxa (pNTX), each one consisting of OTUs sharing pairwise distances not higher than 3%. We found a high diversity and heterogeneity of AMF across the four sites, which showed, in a regression analysis, significant relation to six out of the eight environmental parameters evaluated: grazing activity and soil texture, electrical conductivity, organic matter, total phosphorus and total nitrogen. AMF colonization of plants also presented significant differences among the four sites, as well as spore density, microbial biomass and several enzymatic activities (dehydrogenase, ß-glucosidase and phosphatase) evaluated in rhizosphere soils. The four sites clustered in two groups in a hierarchical clustering evaluation based on their AMF diversity (total numbers of OTU, VTX and pNTX) and the parameters referred above. The crucial role of abiotic factors, other than aridity index, on AMF community composition, was evidenced by the high heterogeneity found between AMF communities across sites under identical aridity conditions.

Composition of the root mycorrhizal community associated with Coffea arabica in Fifa Mountains (Jazan region, Saudi Arabia).

Arbuscular mycorrhizal fungi (AMF) constitute a key functional group of soil biota that can greatly contribute to crop productivity and ecosystem sustainability. They improve nutrient uptake and enhance the ability of plants to cope with abiotic stresses. The presence of AMF in coffee (Coffea arabica L.) plant roots have been reported in several locations but not in Saudi Arabia despite the fact that coffee has been in cultivation here since ancient times. The objective of the present study was to investigate the diversity of AMF communities colonizing the roots of coffee trees growing in two sites of Fifa Mountains (south-west Saudi Arabia): site 1 at 700 m altitude and site 2 at 1400 m. The AMF large subunit rDNA regions (LSU) were subjected to nested PCR, cloning, sequencing, and phylogenetic analysis. Microscopic observations indicated higher mycorrhizal intensity (24.3%) and spore density (256 spores/100 g of soil) in site 2 (higher altitude). Phylogenetic analysis revealed 10 phylotypes, six belonging to the family Glomeraceae, two to Claroideoglomercea, one to Acaulosporaceae and one to Gigasporaceae family. Glomus was the dominant genus at both sites and the genus Gigaspora was detected only at site 2. This is the first study reporting the presence of AMF in coffee roots and the composition of this particular mycorrhizal community in Saudi Arabia.

Characterization of Rhizobial Bacteria Nodulating Astragalus corrugatus and Hippocrepis areolata in Tunisian Arid Soils.

Fifty seven bacterial isolates from root nodules of two spontaneous legumes (Astragalus corrugatus and Hippocrepis areolata) growing in the arid areas of Tunisia were characterized by phenotypic features, 16S rDNA PCR-RFLP and 16S rRNA gene sequencing. Phenotypically, our results indicate that A. corrugatus and H. areolata isolates showed heterogenic responses to the different phenotypic features. All isolates were acid producers, fast growers and all of them used different compounds as sole carbon and nitrogen source. The majority of isolate grew at pHs between 6 and 9, at temperatures up to 40°C and tolerated 3% NaCl concentrations. Phylogenetically, the new isolates were affiliated to four genera Sinorhizobium, Rhizobium, Mesorhizobium and Agrobacterium. About 73% of the isolates were species within the genera Sinorhizobium and Rhizobium. The isolates which failed to nodulate their host plants of origin were associated to Agrobacterium genus (three isolates).

Paenibacillus prosopidis sp. nov., isolated from the nodules of Prosopis farcta.

A bacterial strain, designated PW21(T), was isolated from root nodules of Prosopis farcta in Tunisia. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate into the genus Paenibacillus, with its closest relatives being Paenibacillus glycanilyticus DS-1(T) and Paenibacillus castaneae Ch-32(T) with identity values of 96.9 %. DNA-DNA hybridization measurements showed values of less than 25 % with respect to these two species. The isolate was a Gram-variable, motile and sporulating rod. Catalase activity was positive and oxidase activity was weakly positive. Aesculin, CM-cellulose, xylan and starch were hydrolysed but casein and gelatin were not. Acetoin production was weakly positive and nitrate reduction was negative. Urease production was negative. Growth was supported by many carbohydrates and organic acids as carbon sources. MK-7 was the predominant menaquinone and anteiso-C(15 : 0), iso-C(16 : 0) and iso-C(15 : 0) were the major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, a glycolipid, six phospholipids, an unidentified lipid and two unknown aminophosphoglycolipids. meso-Diaminopimelic acid was not detected in the peptidoglycan. The DNA G+C content of the isolate was 52.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain PW21(T) should be considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus prosopidis sp. nov. is proposed. The type strain is PW21(T) (=LMG 25259(T) =CECT 7506(T) =DSM 22405(T)).