The Effect of Soil and Irrigation Water on The Endophytic Diversity

Abstract

Al -Madinah Mentha ( Mentha longifolia ) is considered one of the most important medicinal and economic plants in Saudi Arabia for its diverse uses in manufacturing perfume, cosmetics, pharmaceuticals and cooking. It is also traditionally used in the treatment of various diseases due to its antimicrobial and antioxidant activity. Endophyte -plant interaction an important role in promoting plant growth and stress tolerance. Plants are capable of selectively choosing their endophytic microbiome to have beneficial associates. Comparative metagenomic analysis benefits in discovering both culturable and unculturable endophytic community different plant tissue.

In this study the endophytic fungal diversity in the different plant parts (leaves and stem) from two different locations in AlMadinah region (Abyar Ali and Abyar Almashi) was determined using metagenomics approach. The most abundant fungal phyla were Ascomycotfollowed by Basidiomycota. All samples are showing high level of species richness. The chemical analysis of irrigation water in Abyar Almashi are showed high value of total Kjeldahl nitrogen. While Abyar Ali water sample has high content of total dissolved solids.

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The essential oil chemical analy sis revealed presence of 21 compounds. The main compound recognized in Abyar Ali mint was carvenone (16.8%followed by 2 -pinene -4-one (14.3%). While in M. longifolia oils from AbyaAlmashi area, 2 -pinene -4-one (13.92%) and pulegone (13.42%) were major components., The results suggest that the endophytic community varies according to the type of irrigation water for each area. For our knowledge this will be the first report discussing this point.

Introduction

Endophytes refer to the endosymbionts microorganisms that asymptomatically colonizing the intercellular and intracellular space of different host plant tissues in a commensal or beneficial relationship.

They commonly include bacteria, fungi and actinomycetes (Kaul, Sharma, Dhar, 2016). Endophyte play a determining role in the plant's health and productivity (Tian, Cao, & Zhang, 2015). The continuous and complex interactions between endophytes and the host plant are a valuable source of novel metabolites and industrially important enzymes (Kaul et al., 2016). The endophytes can produce the same or similar secondary metabolites their host (Heinig, Scholz, & Jennewein, 2013). The host plants can benefit from endophytes with nutrient absorption and improved plant growth as well as biotic and abiotic stress tolerance (Rodriguez et al., 2008). The plant has also benefits from the endophytes by their natural resistance to soil contaminants (Alvin, Miller, & Neilan, 2014). Mentha (mint) is medicinally important plant that belongs to the family Lamiaceae which includes about 42 species that grow extensively worldwide. It can be found in Europe, Africa, Asia, Australia, and North America (Kr?giel et al., 2018). With refreshing flavor and aroma, mint is one of the most popular herbs and it is widely used in cooking, popular medicines, and cosmetics due to many benefits for human health (SaeedNaim, & Tariq, 2006). Mentha species had been used for treatment of many diseases such as ulcerative colitis, throat infection, bronchitis, nausea and liver problem (I?can, Kirimer, K?rkc?o?lu, Ba?er, & Demirci, 2002). Furthermore, it is well -documented that the Mentha species essential oils have strong antioxidant, antifungal, and antibacterial properties against much bacterial growth (de Sousa Barros et al ., 2015).

Mints are one of the most popular herbs in Al -Madinah Al Munawwarah region - Saudi Arabia. Mentha longifolia is the most common species grown in Al -Madinah which is locally known as Al -Madinah mint(locally known as Hassawi and Habaq). The fresh plant is traditionally use as herbal drink by extraction in boiled water as a flavoring agent with hot ea. Also, in traditional medicine it is widely used for treatment of many gastrointestinal disorders (El -badry, Al -ali, & El -badry, 2010) and respiratory disorders. The M. longifolia essential oil showed antioxidant antimicrobial, antispasmodic, and decongestant activities (Iqbal, HussainChatha, Naqvi, & Bokhari, 2013). Most studies done on Mentha species is confined on medical and pharmaceutical aspect, or chemical composition and properties of its essential oils. Very few studies done about Ment ha longifolia in Saudi Arabia. But no published data discuss Mentha microbiome. A metagenomics quite a helpful approach that reveals both culturable and unculturablendophytic microbial communities (Dinsdale et al., 2008). It avoids the need for isolation and cultivation of individual species (Kaul et al., 2016) . Usinmetagenomics in studying the diversity of Mentha microbiome will provide better understanding of Mentha longifolia growth requirements and shall evaluate the high potential of the endophyte community in promoting plant growth and yield and enhancement of plant stress tolerance in different conditions. The Metagenomics studies have improved our understanding of the diversity of microbes in various habitats including plants (AkinsanyaGoh, Lim, & Ting, 2015b). Such studies are important to illuminate endophytes confer stress tolerance and growth promotion to its host plan(Kaul et al., 2016). To our knowledge this is the first report address comprehensive analysis of both cultivable and uncultivable microbe inhabiting this plant.

Literature Review

Since ancient time, the use of medicinal plants has become popular in different parts of the world. It is probably the oldest existing method that humanity has used for treating diseases due to their benefits for human health and their use in food system.

The flora of Saudi Arabia involves about 2250 plant species that are distributed through the kingdom (Collenette, 1999). As one of the most dominate families, Lamiaceae is represented in the flora by 76 species. Of these, 23 species are medical plants (Rahman, Mossa, Al -Said, & Al -Yahya, 2004) . Many of these species are widely growing in Al -Madinah Al - Munawwarah.

Mentha longifolia

The genus Mentha is medicinally important plant that belongs to the family Lamiaceae. Mentha can be classified into 42 species with widespread distribution. Mainly found in Mediterranean region, temperate regions of Europe and eastwards into Asia (Al -Bayati, 2009; Kr?giel et al., 201 8). In Saudi Arabia, they are widely distributed in Wadi Najran, Taif region, and Al Madinah Al Munawwarah (Ibrahim et al., 2017).

The common name mint refers to the aromatic herbal plants with distinct aroma that are cultivated around the world for their essential oils. They have culinary purposes as condiments to add flavor to the food as well as other industrial economic importance. The essential oils form some members of Mentha spp. are used in medicines, pharmaceuticals, cosmetics, pesticide industries, as a flavor enhancing agent in oral hygiene products, chewing gum s and beverages (Croteau, Davis, Ringer, & Wildung, 2005; Shinwari, Sultan, & Mahmood, 2011) .

Isolating bioactive compounds from biological sources such as medicinal plant leads to the discovery of new pharmacologically significant compounds (Alvin et al., 2014) . Studies have been proven that many natural compounds extracted from the essential oil and/or various extracts of Mentha longifolia and other Mentha spp. have shown biological activities such as antifungal, antibacterial, antioxidant, antispasmodic, carminative (Gulluce et al., 2007), antidiarrheal (Jalilzadeh -Amin & Maham, 2015), anti-viral (Amzazi et al., 2003) , anti-cancer, antimutagenic (Al -Ali et al., 2014), The crude extract of M. longifolia had a blood pressure-lowering effect (Shah, Bukhari, & Gilani, 2016). Therefore, this plant has hopeful potential as a medicinal herb to be used for the treatment of various diseases such as bronchitis, flatulence, ulcerative colitis and other liver complaints. (Gulluce et al., 2007), digestive disorders, respiratory disorders and infectious diseases, as well as menstrual disorders in different cultures of all over the world (Farzaei, Bahramsoltani, Ghobadi, Farzaei, & Najafi , 2017). In addition, for relief of minor sore throat, minor mouth or throat irritation and cardiovascular health (Ahmed, Ozbak, & Hemeg, 2015).

Mints are one of the most popular herbs in Al -Madinah Al Munawwarah region - Saudi Arabia. Mentha longifolia is the most common species grown in Al -Madinah which is locally known as Al -Madinah mint (locally known as Hassawi and Habaq). The fresh plant is traditionally used as herbal drink by extraction in boiled water as a flavoring agent with hot tea. Also, in traditional medicine it is widely used for treatment of many gastrointestinal disorders (El -badry, Al -ali, & El -badry, 2010) and respiratory disorders. The M. longifolia essential oil showed antioxidant antimicrobial, antispasmodic, and decongestant activities (Iqbal, HussainChatha, Naqvi, & Bokhari, 2013). Most studies done on Mentha species is confined on medical and pharmaceutical aspect, or chemical composition and properties of its essential oils. Very few studies done about Mentha longifolia in Saudi Arabia. But no published data discuss Mentha microbiome. A metagenomicis quite helpful approach that reveals both culturable and unculturablendophytic microbial communities (Dinsdale et al., 2008). It avoids the need for isolation and cultivation of individual species (Kaul et al., 2016). Usinmetagenomics in studying the diversity of Mentha microbiome will provide better understanding of Mentha longifolia growth requirements and shall evaluate the high potential of the endophyte community in promoting plant growth and yield and enhancement of plant stress tolerance in different conditions. The Metagenomics studies have improved our understanding of the diversity of microbes in various habitats including plants (AkinsanyaGoh, Lim, & Ting, 2015b). Such studies are important to illuminate endophytes confer stress tolerance and growth Mentha spp. are the most popular herbs that widely used in Saudi Arabia. Mentha longifolia is the most common species grown in Al -MadinaAl -Munawwarah region which is locally known as Al -Madinah mintHassawi and Habaq. Large number of researches were performed to support the use of M. longifolia as a traditional popular herbal tea drink both in fresh and dried form. As well as a folk remedy due to many benefits for human health (Farzaei et al., 2017; Mikaili, Mojaverrostami, Moloudizargari, Aghajanshakeri, 2013) .2.1.1 Mentha longifolia Essential OilAmong the various types of compounds, essential oils from aromatic and medicinal plants are receiving particular attention due to their great benefits. Essential oils are very complicated mixtures of natural compounds quite different concentrations (Bozin, Mimica -Dukic, Simin, & Anackov, 2006). Reports showed that many factors interfere with essential oil composition of the different Mentha members. Among these factors, the habitat environment (growth location, soil characteristics), climatical (moisture presence, temperature, etc.), seasonal, and geographical conditions of the regions where the species grow. As well as phenological factors (phase of the plant collection), plant part used for essential oil extraction (flowers, stems, leaves, entire aerial parts or inflorescences), type of material (fresh or dry), and even methods used for essential oil extraction. In addition to environmental stress which can negatively affect the quality of mint essential oils (Hussein, El -Sherbeny, Hendawy, El Gohary, & Ghazal, 2014; Iqbal et al., 2013; Kr?giel et al., 2018).

The variation in the essential oil composition of M. longifolia is reported in the literature from different parts of the world. The major components reported from Bosnia and Herzegovina were piperitone oxide and 1,8 -cineole (Nik?i?, Kova? -Be?ovi?, Makarevi?, & Duri?, 2012). Oil obtained from Turkey have cis -epoxy piperitone, pulegon e and piperitenone oxide as main components (Gulluce et al., 2007). Moroccan Mentha Longifolia essential oil showed unusual relevant amounts of piperitenone oxide and piperitone oxide (Ghoulami, II Idrissi, & Fkih -Tetouani, 2001). Samples of essential oils from Tunisia were characterized by a high amount of 1,8 -cineole, menthone and terpineol 1 -4 (Hajlaoui, Snoussi, Ben Jannet, Mighri, & Bakhrouf, 2008). Oil obtained from Iranian collection revealed that pulegone was the major component followed by 1,8 -cineole and sabinene (Golparvar & Hadipanah, 2016). The major constituents of the essential oil of M. longifolia collected from Pakistan were piperitenone oxide, piperitenone and germacrene D (Iqbal et al., 2013). The main compounds in M. longifolia oil from Belgrade were trans - and cis - dihydrocarvone and piperitone followed by 1,8 -cineole (D?ami? et al., 2010) .

Irrigation Water

The waters used for irrigation contain appreciable quantities of unnecessary or unwanted substances which are dissolved in the water from the geological strata which the waters have flowed through and over or manmade (domestic and industrial effluents) sources. The substance severity depends upon its type and its physiochemical characteristics. The main water sources in Saudi Arabia are underground water aquifers, rainfall and seawater desalination. Since there are no perennial streams or rivers quantity (Hussain, Alquwaizan, & Al -Zarah, 2010). Aquifers and individual wells can be contaminated by substantially different processes. However, the suitability of water for irrigation depends on the physical, chemical and biological characteristics of water. The application of poor or hazardous quality water may affect the agriculture soil fertility, crop growth and reduce the crop yield in several ways (Arshad & Shakoor, 2011).

The Chemical Quality of Irrigation Water

Different organizations have established standard guidelines for judging the suitability of water for irrigation. One of the most common water quality indices/parameters that describe the quality of water is the Total Dissolved Solids (TDS). Salinity is a common problem facing farmers who irrigate in arid climates. The salinity behavior of water is indicated by its TDS concentration (Arshad & Shakoor, 2011). The salt concentration in most irrigation waters ranges from 200 to 4000 mg /liter total dissolved solids (Andreas Phocaides, 2007). The application of irrigation water to the soil introduces salts into the root zone. Therefore, the soil salinity increases in direct proportion to the salinity of irrigation water. The accumulation of salts in the root zone increases the osmotic pressure of the soil solution and limits the water availability for the plant which results in decreased crops yield (Hussain et al., 2010) . According to the Food and Agriculture Organization (FAO), irrigation water quality was classified to five categories in accordance with their salinity level as shown in Table 1 (Ayers & Wescott, 1994).