Flame photometer 128

The soil pH and conductivity were determined in 1:5 soil:water suspension using HM digital meter-COM-100 (New Delhi, India) and Equip-tronics EQ-614-A (Mumbai, India), respectively. The soil suspension was prepared by mixing soil and water in desired proportion (1:5) and the mixture was shaken for 2 h and the supernatant was filtered and used for measurement. Soil texture and organic carbon content was determined by Hydrometer method (Jacob and Clarke 2002 ) and Walkley–Black wet oxidation method (Nelson and Sommers 1982 ) respectively. A factor of 1.72 was multiplied with organic carbon content to determine SOM. EDTA titration method was used for measuring calcium (Ca) and magnesium (Mg) (Lanyon and Heald 1982 ), acid neutralization method for CaCO₃ (Hesse 1971 ) and potassium (K) and sodium (Na) were measured by using a Systronics Flame Photometer-128, after digesting the samples in a diacid mixture (HClO4/HNO3 in a 4:1 ratio) (Bhat et al. 2014 (link)). Total nitrogen (N) was determined by Kjeldahl method (Bremner and Mulvaney 1982 ) and available phosphorous (P) by sodium bicarbonate extraction method using Spectrophotometer-2202 (Ahmedabad, Gujarat, India) (Olsen et al. 1954 ).

Na⁺ and K⁺ contents of leaves, roots, and bulbs were determined at the time of harvesting using the flame photometer (Systronics Flame Photometer 128, Olathe, KS, USA) and estimated as mg g⁻¹ of dry weight.

Sodium (Na) and potassium (K) were determined using flame photometer 128 (Systronics), phosphorus (P) by using spectrophotometer DR5000 (HACH). Calcium (Ca) and magnesium (Mg) were determined by using Atomic Absorption spectrophotometer (AnalytiKjena ZEEnit 700, Germany). Other mineral elements namely, iron, copper, zinc, manganese, molybdenum, nickel, lead, cadmium, cobalt and arsenic were determined using ICP-Mass Spectrophotometer (NexIon^® 300X Series, Perkin-Elmer). All the standards of different mineral elements used in the study were of analytical grade. All the observations were recorded in two replications.

TY and EV yeast strains were grown in 10 mL of LSM supplemented with galactose, or in the treatment media (LSM) supplemented with galactose and 25 mM NaCl for 48 h at 30 °C and shaken at 200 rpm in a shaking incubator. To extract Na⁺ and K⁺ from yeast cells, cells were collected in 2 -mL tubes by centrifugation at 5000 rpm for 5 min before being washed three times with sterile distilled water. Finally, the yeast cells were suspended in 1 mL sterile distilled water and boiled at 100 °C for 20 min in a water bath. After boiling the tubes, they were centrifuged at 12,000 rpm for 10 min, with the clear supernatant separated and diluted 10 times for Na⁺ and K⁺ measurements using Systronics Flame Photometer 128 (Systronics India Ltd. Ahmedabad, Gujarat, India) as previously described [26 (link)].
Arabidopsis plant samples were prepared by digesting the plant tissue using weak nitric acid as previously described by Munns et al. [27 ]. Briefly, plant samples were dried, weighted and digested in 10 mL of a 0.1 M nitric acid solution. Subsequently, the samples were kept at room temperature and shaken for two days at 100 rpm before being filtered using Whatman No. 1 filter paper and used for Na⁺ and K⁺ measurements. The Na⁺ and K⁺ concentrations were measured as previously mentioned in this study.

Fresh leaf samples and flower buds were collected one month before flower bud differentiation and at the time of fruit bud differentiation, respectively. For this, terminal shoots were tagged in July, and leaves were harvested after 4 months of growth for recording biochemical traits and nutrient acquisition, while buds were collected in December. A total of 30 leaves /samples (3^rd and 4^th mature leaves from the top of the terminal shoot) were collected for sampling for nutrient observations. For estimation of P, K, Na, Ca, Mg, S, and micronutrients B, Fe, Mn, Cu, and Zn, one gram of the plant sample was digested in 4:1 nitric acid and perchloric acid (HNO₃: HClO₄) mixture. Phosphorus (%) (Vanadomolybdophosphoric yellow color method) [25 ] with spectrophotometer (Systronics UV- VS Spectrophotometer 117), Potassium (%) and Sodium with Flame photometer (Systronics Flame Photometer 128) [25 ] and Sulphur (%) (Turbidimetric method) [26 (link)] with spectrophotometer (Systronics UV- VS Spectrophotometer 117) estimated. Calcium (%), Magnesium (%) [25 ], and micronutrients- Fe, Mn, Cu, Zn (ppm) [27 (link)] with Motras Scientific Atomic Absorption Spectrophotometer recorded. Boron (ppm) estimated with Azomethine-H [28 (link)] with spectrophotometer- [Systronics UV- VS Spectrophotometer 117].

pH and electrical conductivity (EC) of initial feed mixture and final products were measured in distilled water suspension of each concentration in the ratio of 1:10 (W/V) using Systronics μ pH system 362 and Systronics conductivity meter-304, respectively. Total organic carbon (TOC) was determined after burning the 0.5 g of waste in a muffle furnace at 550°c for 60 min as described by Nelson and Sommers (1996 ). Micro-Kjeldhal method of AOAC (2000 ) was used for measuring nitrogen after digesting the waste in digestion mixture (H₂SO₄ + K₂SO₄:CuSO₄:SeO₂ in 10:4:1). The method described by John (1970 (link)) was used for measuring total available phosphorus (TAP) using Systronics double beam spectrophotometer 2202, total potassium (TK) and sodium (TNa) was measured by using a Systronics flame photometer-128 after digesting the samples in diacid mixture (HClO₄:HNO₃ in 4:1 ratio). Heavy metals were measured by Agilent 240 FS AA model Atomic Absorption Spectrophotometer in the digested samples.

The soil samples were analyzed for texture, total dissolved solids (TDS), pH, electrical conductivity (EC), organic carbon (OC), nitrogen (N), sodium (Na), calcium (Ca), phosphorus (P), potassium (K). The method of Bouyoucos [42 (link)] as used for soil texture analysis. The content of pH, EC, and TDS was analyzed by making the suspension of 1:10 (soil sample: distilled water) using a shaker and finally measured with a digital meter (Eutech Instruments). The OC was measured by using Nelson and Sommers [43 ] method after igniting the soil samples in a muffle furnace at 550 °C. The N was analyzed according to Bremner and Mulvaney [44 ] method after digesting the soil sample with concentrated H₂SO₄ followed by running the sample in Kjeldahl assembly and finally titration was done with 0.01 N HCl. The process of John [45 (link)] was used for phosphorus estimation after digesting the soil with 1:4 of perchloric acid and nitric acid respectively followed by using a spectrophotometer (Systronics). The content of Na, K and Ca was analyzed from samples digested for phosphorus by using Flame Photometer-128 (Systronics).