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.

Fusarium oxysporum and Colletotrichum musae Associated with Wilt Disease of Coffea arabica in Coffee Gardens in Saudi Arabia.

This study aimed to identify if Fusarium and Colletotrichum species are linked to coffee leaf wilt symptoms (Coffea arabica L.) gardens in Jazan region, Kingdom Saudi Arabia. The symptomatic wilted leaves and shoots were collected from Jazan Mountain Region Development Authority (JMRDA) farm in jabal Fifa. Samples of roots and leaves tissues were plated on Dox' Agar medium and incubated for one week at 24oC. Two morphologically different fungus colonies grew on the medium. A PCR-based method was used for the molecular amplification and characterization of the fungi using a 18SrRNA specific primer. 1323 and 1501 bp PCR products were obtained by using the 1% agarose gel electrophoresis. The sequence analysis and genbank homology revealed that the present fungi were Fusarium oxysporum and Colletotrichum musae with 99 and 98% similarity, respectively. Both fungi sequences were submitted to the genebank under accession numbers OP010081 and OP010082, respectively. This is the first report of these two genera of fungi infecting the roots and leaves of coffee trees in Jazan Region of Saudi Arabia and suggests that other fungus species may play a significant role as diseases in other coffee-producing areas.

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.

Salvastearolide, a new acyl-glyceride, and other constituents from the seeds of Salvadora persica

Abstract The chemical investigation of the n-hexane fraction of Salvadora persica L., Salvadoraceae, seeds afforded a new stearic acid ester, salvastearolide, together with five other phytosteroids identified as stigmasterol, β-sitosterol, Δ7-campesterol, Δ7-avenasterol and campesterol. Their structures were established on the basis of extensive spectroscopic methods including 1D and 2D NMR experiments and HRESI mass spectrometry. In addition, salvastearolide and the isolated fractions were tested for their cytotoxicity against human cancer cell lines MCF-7, MDA-MB-231 and HT-29. The n-hexane fraction exhibited significant anti-proliferative effect against human breast cancer cell line MCF-7 (IC50 50 µg/ml), while salvastearolide possessed a weak cytotoxic effect against MCF-7 cells with IC50 103.98 µg/ml.

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.

NaCl stress-induced changes in the essential oil quality and abietane diterpene yield and composition in common sage.

AIM: The purpose of this study was to evaluate how increasing NaCl salinity in the medium can affects the essential oils (EOs) composition and phenolic diterpene content and yield in leaves of Salvia officinalis L. The protective role of such compounds against NaCl stress was also argued with regard to some physiological characteristics of the plant (water and ionic relations as well as the leaf gas exchanges). MATERIALS AND METHODS: Potted plants were exposed to increasing NaCl concentrations (0, 50, 75, and 100 mM) for 4 weeks during July 2012. Replicates from each treatment were harvested after 0, 2, 3, and 4 weeks of adding salt to perform physiological measurements and biochemical analysis. RESULTS: Sage EOs were rich in manool, viridiflorol, camphor, and borneol. Irrigation with a solution containing 100 mM NaCl for 4 weeks increased considerably 1.8-cineole, camphor and ß-thujone concentrations, whereas lower concentrations (50 and 75 mM) had no effects. On the contrary, borneol and viridiflorol concentrations decreased significantly under the former treatment while manool and total fatty acid concentrations were not affected. Leaf extracts also contained several diterpenes such as carnosic acid (CA), carnosol, and 12-O-methoxy carnosic acid (MCA). The concentrations and total contents of CA and MCA increased after 3 weeks of irrigation with 75 or 100 mM NaCl. The 50 mM NaCl had no effect on these diterpenes. Our results suggest a protective role for CA against salinity stress. CONCLUSION: This study may provide ways to manipulate the concentration and yield of some phenolic diterpenes and EOs in sage. In fact, soil salinity may favor a directional production of particular components of interest.

Possible involvement of polyphenols and polyamines in salt tolerance of almond rootstocks.

Leaf physiological and biochemical adaptive strategies and more particularly the possible involvement of polyamines and polyphenols in salt stress tolerance were investigated. Three almond rootstocks (GN15, GF677 and bitter almond) were subjected to 0, 25, 50 and 75 mM NaCl for 30 days. The dry mass of leaves, stems and roots decreased with increasing salt concentration in the irrigation solution regardless of genotype. Photosynthetic assimilation rate decreased in the three almond rootstocks, but more so in GF677 and bitter almond. The accumulation of toxic ions was greater in the leaves than in the roots in all genotypes. GN15 accumulated less Na(+) and Cl(-) than GF677 and bitter almond. GF677 accumulated polyphenols, but had less anthocyanin and antioxidant activity in its leaves compared to bitter almond. It seems that GN15 was more able to tolerate the excess of toxic ions using anthocyanins which are abundant in its red leaves and free polyamines for a more efficient response to stress. However, most of the antioxidant activity was found in the leaves and was lower in the roots. Given that the upper part of the tree will be of a different cultivar after grafting, this advantage may not be relevant for the tree's survival. GF677 showed a different antioxidant strategy; it maintained a stable carotenoids content and accumulated polyphenols in its leaves. The three rootstocks used different strategies to deal with the excess of salt in the growth medium.

Kinetin applications alleviate salt stress and improve the antioxidant composition of leaf extracts in Salvia officinalis.

A pot experiment was carried out under glasshouse conditions with common sage (Salvia officinalis L.) to investigate the interactive effects of salt stress and kinetin on growth attributes and the abundance of pigments, ions, phenolic diterpenes and α-tocopherol in leaf extracts of this species. The plants were subjected to the following four treatments: (i) control (nutrient solution), (ii) control + 10 µM kinetin, (iii) salt stress (nutrient solution + 100 mM NaCl), and (iv) salt stress + 10 µM kinetin. Kinetin was applied as a foliar fertilizer. Salt stress reduced water contents, photosynthetic activity and pigment contents of sage leaves. In addition, it increased Na(+) contents, and reduced those of Ca(2+) and K(+) in leaves. Salt stress reduced carnosic acid and 12-O-methyl carnosic acid contents in leaves, while it did not affect carnosol and α-tocopherol contents. Foliar applications of kinetin seemed to counterbalance or alleviate the stress symptoms induced by salinity, improving ion and pigment contents, while leaf phenolic diterpene (mainly carnosol) and α-tocopherol contents also increased in both control and NaCl-treated plants; still this effect was much more obvious in salt-treated plants. A similar effect was also obtained when plants were sprayed with KNO(3) or Ca(NO(3))(2), thus suggesting that kinetin effects were at least partly due to an improvement of ion homeostasis. Kinetin applications resulted in increased transcript levels of the isoprenoid and tocopherol biosynthetic genes, DXPRI and VTE2 and VTE4 in control plants, but not in NaCl-treated plants. We conclude that kinetin can alleviate the negative impact of salt on sage plants cultivated under arid environments with salinity problems.

How reliable is the double-ended pressure sleeve technique for assessing xylem vulnerability to cavitation in woody angiosperms?

The reliability of a double-ended pressure sleeve technique was evaluated on three woody angiosperm species with contrasting maximum vessel lengths. Vulnerability curves (VCs) were constructed by varying sample length and the size of the pressure sleeves. VCs were compared against curves obtained with reference techniques. For the two diffuse-porous species, Betula pendula and Prunus persica, VCs built with shoot segments shorter than maximum vessel length strongly overestimated species vulnerability. Furthermore, increasing the size of the pressure sleeve also tended to lead to overestimated VCs. For the ring-porous species Quercus robur, the technique strongly overestimated vulnerability to embolism, whatever the sample length or chamber tested. In conclusion, the double-ended pressure sleeve technique only gives reliable VCs on diffuse-porous angiosperms with short pressure sleeves, only when segments are longer than maximum vessel length.

Influence of ionic interactions on essential oil and phenolic diterpene composition of Dalmatian sage (Salvia officinalis L.).

The potential of four essential cations (K(+), Ca(2+), Mg(2+) and Fe(2+)) to alleviate salt toxicity was studied in sage (Salvia officinalis L.) plants grown in pots. Two concentrations of the following chloride salts: KCl, CaCl2, MgCl2 and FeCl3, were used together with 100mM NaCl to study the effects of these nutrients on plant growth, leaf essential oils (EOs) and phenolic diterpenes composition. The sage plants accumulated Na(+) in their leaves (includers); this has affected secondary metabolites' biosynthesis. Treatment with 100mM NaCl slightly decreased borneol and viridiflorol, while increased manool concentrations. Addition of KCl, CaCl2 and MgCl2 increased considerably in a dose-dependent manner the oxygen-containing monoterpenes (1.8-cineole, camphor, ß-thujone and borneol) in 100mM NaCl-treated sage. Whereas, the contents of viridiflorol decreased further with the addition of KCl in 100mM NaCl-treated sage. Our results suggest that the changes in EOs composition were more related to K(+) and Ca(2+) availability than to Na(+) toxicity. Furthermore, treatment with NaCl decreased by 50% carnosic acid (CA), a potent antioxidant, content in the leaves. K(+) and Ca(2+) promoted the accumulation of CA and its methoxylated form (MCA) in the leaves. The concentration of CA was positively correlated with leaf K(+) (r=0.56, P=0.01) and Ca(2+) (r=0.44, P=0.05) contents. It appears that different salt applications in combination with NaCl treatments had a profound effect on EOs and phenolic diterpene composition in sage. Therefore, ionic interactions may be carefully considered in the cultivation of this species to get the desired concentrations of these secondary metabolites in leaf extracts.

Water relations and drought-induced embolism in olive (Olea europaea) varieties 'Meski' and 'Chemlali' during severe drought.

We examined the effects of drought on the water relations, osmotic adjustment and xylem vulnerability to embolism of olive (Olea europaea L.) varieties, 'Meski' and 'Chemlali'. Two-year-old self-rooted cuttings growing in sand-filled pots in a greenhouse were subjected to water stress by withholding water for 60 days. Water relations and gas exchange measurements showed that 'Chemlali' was more drought resistant than 'Meski' and had a greater capacity for osmotic adjustment through solute accumulation. However, when water stress was acute, the effect of osmoregulation on leaf cell turgor was largely counteracted by xylem cavitation. Cavitation vulnerability curves showed that both varieties were highly resistant to embolism formation. The xylem water potential inducing 50% loss of stem conductivity approached -7 MPa in 'Meski' and only slightly less in 'Chemlali'. Although the difference between varieties in susceptibility to xylem embolism was small, it appears to account in large part for the difference between them in the ability to tolerate severe drought.

The effect of sodium chloride salinity on the growth, water status and ion content of Phragmites communis trin.

The present study deal with the physiological behavior of Phragmites communis under salt stress. The effects of salinity on growth, dry weight partitioning, water status and ion content were studied on seedlings of P. communis fed with nutrient solutions containing 0 to 600 mM NaCl. The plants grew best when irrigated with distilled water; biomass production and Relative Growth Rate (RGR) decreased with increasing salinity. Nevertheless, plants were able to produce and allocate dried matter to all their organs even at the highest salt level (600 mM NaCl). The leaves showed the lowest growth activity. Increasing salinity was accompanied by a decrease in seedling water content; aerial parts were more dehydrated than roots. Examination of the K+/Na+ selectivity revealed that salt tolerance of reed plants may be due to its capacity to limit Na+ transport and to enhance K+ transport into aerial parts resulting in a high K/Na ratio. Our results suggest an exclusive behavior towards Na+ as shown by the decreasing Na+ gradients from leaves to roots. It is concluded that Na+ exclusion mechanism appeared to be operative and contributes to salt tolerance of Phragmites.