Effect of Some Fertilization Treatments on Vegetative Growth, Oil Production and Chemical Composition of Sage Plant

INTRODUCTION

Salvia officinalis L. (sage) is a small perennial evergreen subshrub, with woody stems, grayish leaves, and blue to purplish flowers. It is a member of the mint family, Lamiaceae. It is native to the Mediterranean region and commonly grown as a kitchen and medicinal herb or as an ornamental garden plant. The principal constituents of essential oil of sage are thujone (about 42%), cineole, borneol, caryophyllene and other terpene(Lowes, 1992). Although the effectiveness of common sage is open to debate, it has been recommended at one time or another for virtually every ailment. Modern evidence supports its effects as an anhydrotic, antibiotic, antifungal, astringent, antispasmodic, estrogenic, hypoglycemic, and tonic. In a double blind, randomized and placebo-controlled trial, sage was found to be effective in the management of mild to moderate Alzheimer's disease (Akhondzadeh et al., 2003).

Plant nutrition is one of the most important factors affecting quantity and quality of secondary metabolites in plants. In order to meet the ever increasing demand of medicinal plants need to be identified the best fertilizer application strategies. N, P and K are essential macronutrients for plant growth and yield (El-Hag, 2001). Because, they partake in many components structure i.e. carbohydrates, protein, phospholipids, amino acids, nucleic acids etc. (Devlin, 1975). Ascorbic acid plays a role in plant growth and development, cell division, cell wall metabolism and cell expansion, shoot apical meristem formation, root development, photosynthesis, regulation of florescence and regulation of leaf senescence. Also, it is cofactors for enzyme activity, and effects on plant antioxidation capacity, heavy metal evacuation and detoxification and stress defense (Zhang, 2012).Citric acid prevents the reaction of free radical, with biomolecules and can remind the nutritional values and physiological properties of food stuffs (Rasmy   et al., 2012).The effect of  active dry yeast is due to its capability in induction of endogenous hormones like GA₃ , IAA and cytokinins (Khedr and Farid, 2000), as well as, dry yeast a natural source of vitamin B group, amino acids and nutritional elements, protein, carbohydrate, nuclic acid and lipids (Nagodawithana, 1991).Therefore, the aim of the study was to investigate the effects of (NPK), ascorbic and citric acids and active dry yeast on sage. As well as, to find out the best fertilization treatment to improve the growth, chemical composition and oil productivity of sage and reducing the intensive use of chemical fertilizers.

MATERIALS AND METHODS

Field experiment was conducted at the private Farm at the Al-Delingat- Al-Bhyra gavernorate, Egypt during two successive seasons (2014 and 2015) to study the Effect of (NPK), active dry yeast, ascorbic and citric acids on vegetative growth, essential oil productivity and chemical composition of sage (Salvia officinalis, L.) plants.Seeds of sage were sown on 15^th January in germination dishes and were putted under plastic house of especial nursery in 2014 and 2015 seasons. Seedlings of 12 cm height with 8-10 leaves were cultivated on 15^th and  18^th  March in the both seasons, respectively in plots of 4m² (2×2m) with 30 cm between plants and 50 cm between rows, (4 row in plot) each plot was planted with 28 plants. The physical and chemical analyses of the used soil are shown inTable (1) according to Page et al. (1982) and Klute (1986).

Table (1). Some physical and chemical analysis of the used soil
                     before planting (average two seasons of 2014 and 2015).

Fertilizer types:

1. Cattle manure at the rate of 10 kg/plot was added at the soil preparation for all treatments in the two seasons, its analysis as shown in Table (2).

 Table (2). Analysis report of cattle manure

2. Ammonium sulphate (20.5 % N), calcium superphosphate (15.5 %P₂O₅) and potassium sulphate (48 % K₂O) at the rates of 10.72, 7.14 and 3.57 g/Plant (300+200+100 kg/fed), respectively, as recommended dose (Shala, 2007),whereas, calcium superphosphate was added at one dose before planting at soil preparation, while, ammonium sulphate and potassium sulphate were divided into four equal doses. The first one was done after 20 days from transplanting; the second one was done after 50 days from transplanting, the third dose after 15 days from the first cut and the fourth dose after 35 days from the first cut.

3. Active dry yeast (ADY) at the rates of 3 and 4 g/l was sprayed 4 times after 25 and 55 days after transplanting and 20 and 40 days after the first cut for both seasons.

4. Ascorbic and citric acids (AA and CA) at the rates of 200 and 250 mg/l for each were sprayed 4 times after 22 and 52 days after transplanting and 18 and 37 days after the first cut for both seasons. The spraying of ADY, AA and CA was done in morning tell run off.

The fertilization treatments were conducted as follow:

T1= NPK recommended dose (control), T2= 3/4 NPK dose +3g /l ADY, T3= 3/4 NPK dose +200 mg/l CA, T4= 3/4 NPK dose +200 mg/l AA, T5= 1/2 NPK dose +3g /l ADY +200 mg/l CA, T6= 1/2 NPK dose +3g /l ADY +200 mg/l AA, T7= 1/2 NPK dose +200 mg/l of each CA and AA, T8= 1/4 NPK dose +200 mg/l of each CA and AA +3g /l ADY. T9= 1/4 NPK dose +250 mg/l of each CA and AA and T10= 250 mg/l of each CA and  AA + 4g /l ADY.

The experiment was designed as randomized complete block design with three replicates. Each replicate contained 10 treatments, one plot (2x2 m) as an experiment al unit.

At the first and second cuts on 2^nd August and 2^nd Dec., respectively for the two seasons, the following data were recorded:

1. Vegetative growth traits (plant height (cm), branches number/ plant and fresh and dry weights (g) of aerial parts/plant.

2. Essential oil % was determined in the air dried herbs according to British Pharmacopoeia (1963), by the formula of Aliabadi et al. (2008).

Essential oil % =(Oil volume in graduate tube / Sample weight (g)) X 10

Essential oil yield/ plant= Essential oil % x plant dry weight

3. Chemical analysis (leaf green color degree was done 3 days before each cut as SPAD units using "Minolta (chlorophyll meter) SPAD-502 (Yadava, 1986), N % according to method of Chapman and Pratt, (1978), P and K according to Jackson (1973) and carbohydrates % using methods of Yemm and Willis (1954) in leaves.

Data were subjected to statistical analysis of variance as described by Gomez and Gomez (1984). The treatment means were compared using L.S.D. test at 0.05 level of significant.

RESULTS AND DISCUSSION

1. Effect of fertilization treatments on vegetative growth of sage plants:

Data in Table (3) cleared that the used different fertilization treatments significantly affected vegetative growth traits of sage during the two cuts in the two seasons, whereas, the significantly tallest plants and the highest shoots number per plant were recorded for the plants received NPK full dose (control), besides of T4 in case of plant height in the two cuts during the both seasons, without significant difference between them, except for plant height in the first cut during the second seasons. On the other side, the significantly shortest plants and minimum shoots number/plant resulted from applying T7 and T8, respectively, during the two cuts in the both seasons. Also, data in Table (3)showed that the significantly highest values of aerial parts fresh and dry weights were achieved from the treatment of 3/4 NPK dose + 200 mg/l ascorbic acid (T4) for the two cuts in the both seasons, while the significantly least values of plant aerial parts fresh and dry weights resulted from T9 for the two cuts in the two seasons, plus T2 and T5 in case of fresh weight of aerial parts in the second cut in the second season, with one exception in case of plant aerial parts fresh weight during the first cut in the second season which resulted from T10.

The treatment of 3/4 NPK dose + 200 mg/l ascorbic acid achieved the best results for plant height and fresh and dry weights of plant aerial parts, besides of NPK full dose in case of plant height and shoots number may be due to the important role of N, P and K elements in many physiological and biological processes and partake in many components in plant (Devlin, 1975), as well as, the role of ascorbic acid in cell division, cell wall metabolism and cell expansion, shoot apical meristem formation, root development, photosynthesis and regulation of leaf senescence ,also, it is cofactors for enzyme activity and effects on plant antioxidation capacity, heavy metal evacuation and detoxification and stress defense (Zhang, 2012), all that reflected on plant height, shoots number and fresh and dry weights of plants.These results are in agreement with those of Boroomand et al. (2012) who concluded that N, P and K elements caused increases in traits such as plant height, leaf area and yield seed of basil, turmeric, black pepper, cardamom and fennel. Also, Abdoua and Mohamada (2014) found that 48 t/fed plant compost, 150 mg/l ascorbic acid and 150 mg/l salicylic acid yielded fresh and dry weights, of Mentha piperita higher than NPK fertilizer. They added that plant compost, ascorbic acid and salicylic acid effectively improved mint productivity, and can reduce the use of chemical fertilizers and conserve natural resources.

Table (3). Effect of fertilization treatments on some vegetative growth of sage (Salvia officinalis, L.)during the two cuts in seasons of 2014 and 2015.

2. Effect of fertilization treatments on oil productivity of sage plants:

Data in Table (4) revealed that plants fertilized by T10 during the two cuts in the both seasons, in addition to those received T7, T8 and T9 in the second cut in the first season had the significantly highest oil percentage. On the opposite the plants received T3 during the both cuts in the two seasons in addition to those received T2 during the second cut in the both seasons had the least significant essential oil percentage in comparison to the other treatments.

Also, data in Table (4) demonstrated that the significantly highest essential oil yield during the two cuts in the both seasons was achieved from the plants fertilized with T4 (3/4 NPK dose + 200 mg/l ascorbic acid), but the least significant essential oil yield per plant resulted from T9 during the two cuts in the first season and from T2 during the two cuts in the second one.

The treatment of 250 mg/l of each citric acid and ascorbic acid + 4 g/l active dry yeast gave the beast essential oil % may be attributed to the several types of biological activities in plants such as enzyme cofactors and antioxidant which associated with ascorbic acid (Conklin, 2001), also, ascorbic acid enhance plant growth and development and plays a role in electron transport system (El-Kobisy et al., 2005), furthermore, citric acid appears as an intermediate in the basic physiological citric acid or Krebs cycle in every eukaryote cell (Karlaganis, 2001), in addition to the role of active dry yeast in improving many physiological and biochemical processes as a result of its contain from macro and micronutrients, amino acids, protein, sugars, organic acids and cytokinins (Kurtzman and Fell, 2005 and Ezz-El-Din and Hendawy, 2010), all that reflected on the metabolic processes, it turn account on essential oil percentage.

As well as, the treatment of 3/4 NPK dose + 200 mg/l ascorbic acid resulted in the highest value of essential oil yield per plant may be due to the enhancing effect of essential elements (N, P and K) and ascorbic acid on the plant growth , it turn account on plant dry weight consequentily more essential oil yield.

These results are in accordance to those of Mahgoub (2009) concluded that spraying Melissa officinalis with active dry yeast caused a significant increase in essential oil yield. Likewise, Khalil et al. (2010) who indicated that application of ascorbic acid in different concentrations showed significant increases in oil % of basil. The highest essential oil yield of bail was observed in citric acid 0.1 % ( w/v), citric acid ease of availability and usage makes it a promising candidate in manipulation of secondary metabolism related pathway in medicinal plants (Jaafari and Hadavi, 2012). Also, maximum essential oil value of Ajowan was gained from 200 kg N/ha (Vahidipour et al., 2013).

Table (4). Effect of fertilization treatments on essential oil percentage and oil yield per plant (g) of sage (Salvia officinalis L.) during the two cuts in seasons of 2014 and 2015.

3. Effect of fertilization treatments on leaf chemical composition of sage plants.

Data in Table (5) cleared that fertilization treatments significantly affected chemical traits during the two cuts in the both seasons. Whereas, the significantly highest values of leaf green color degree and the percentages of N, K and total carbohydrate during the two cuts in the both seasons resulted from applying T4, in addition to T2, T8 and T10 at the second cut in the first season in case of leaf green color degree and T6 at the two cuts in the first season in case of total carbohydrate %. While highest significant P % was recorded for T1, T8 and T10 during the two cuts in the both seasons, besides some treatments during different cuts in the two seasons, without significant differences among them.

On the other side, the significantly least values of leaf green color degree resulted from T7 during the two cuts in the both seasons, N% from T5 and T7 in the two cuts for both seasons, except for the second cut in the second season, P% from T5 during the two cuts in the both seasons, K% from T5 during the two cuts in the both seasons, besides T2 at the first and second cuts in the both seasons, respectively, and T3 at the two cuts in the first season and total carbohydrate % from T1 during the two cuts in the two seasons. The other used fertilization treatments gave intermediate values for such parameters with significant differences among themselves in the most cases during the two cuts in the both seasons.

The treatment of 3/4 NPK dose +200 mg/l ascorbic acid (T4) improved the chemical traits may be due to the increase essential elements (N, P and K) in the root zone which reflected on its uptake by plant roots and it turn account on photosynthesis, N, P, K and total carbohydrate in plant leaves (Devlin, 1975), because of these elements play important roles in many physiological and biochemical processes in the plant (Wu et al., 1998, Abel et al.,  2002 and Bashir, 2012). In addition, to the role of ascorbic acid that might   increase the organic acids excreted from the roots into the soil and consequently increases the solubility of most nutrients which release slowly into the rhizosphere zone where it may be utilized by the plants, also, ascorbic acid acts as coenzyme reactions by which carbohydrates, facts and protein were metabolized (Ahmed, 1996).

These results are similar to those of Rupa et al. (2007) who concluded that maximum chlorophyll content and dry matter production were obtained under 5 foliar sprays of  NPK at 17+ 10 +27, Mazher et al. (2011) revealed that ascorbic acid at 200 mg/l increased total carbohydrates, N, P and K% in Codiaeum variegatum, Mosleh et al. (2014) indicated that chlorophyll content in the apricot leaves significantly increased by increasing sheep manure, ascorbic acid and sulphur levels and El-Morsy (2015) mentioned that application of  12+ 6+ 6g/ plant NPK plus 5% compost from soil dry weight in pot for Duranta and Murraya seedlings resulted in the beast values of green color degree and total carbohydrates.

Table (5). Effect of fertilization treatments on chemical compositions of sage (Salvia officinalis, L.) during the two cuts in seasons of 2014 and 2015.

Conclussion

Results cleared that to achieve the beast vegetative growth, oil production and chemical composition it can be fertilized Salvia officinalis with 225 kg/fed ammonium sulphate + 150 kg/fed calcium superphosphate + 75 kg/fed potassium sulphate + 200 mg/l ascorbic acid. Whereas, calcium superphosphate is added as one dose at soil preparation before planting, while, ammonium sulphate and potassium sulphate are divided into four equal doses, where the first and second doses are done after 20 and 50 days from transplanting, respectively, and the third and fourth  doses are added after 15 and 35 days from the first cut, respectively. While, ascorbic acid is sprayed four times at 22 and 52 days from transplanting and 18 and 38 days after the first cut.