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India occupies a special status in terms of ecosystem, species and genetic diversity because of its location in the tropical zone, physical features and eco-climatic conditions. Rajasthan is the largest State of India and is situated in the north-western part of India between 23°3’N and 30°12’N latitude and 69°30E and 78°17’E longitude, occupying an area of 3,42,239 sq.km. The elevation of land surface varies from 214 to 1375 m. In shape, it is an irregular rhomb with north-south and east-west diagonals, the former about 784 km. and the latter 850 km. long.[9] The remarkable feature of Rajasthan is the Aravalli range, perhaps the oldest folded mountain range in the world. It intersects Rajasthan from end to end, diagonally running from Delhi to the plains of Gujarat for a distance of about 692 km. Within Rajasthan, the range runs from Khetri in the north-east to Khed Brahma in the south- west for a length of about 550 km. The elevation of the Aravalli range gradually rises in south-west direction, as it is 335 m at Delhi and in Rajasthan 792 m at Khetri, and 1727 m at Mt. Abu. Further south- west wards, the elevation gradually decreases to the plains in Gujarat. It has a wide range of habitats, climatic factors, physiography, soil types, and geological antiquity. Phytogeographically, the state of Rajasthan forms the eastern extremity of the great arid and semi-arid belt of the world. Nagaur district is located in the center of the Rajasthan state between 26°25' and 27°40' north latitudes and 73°10' and 75°15' east longitudes. This district covers an area of 17,718 sq. km. which is about 5.177% of the total area of the state. The present study is a report based on a survey of Angiospermic plants of Deedwana block of Nagaur district over five years. Regular and periodical visits to different habitats were made during these years of intensive survey. A total of three hundred thirty-one species, grouped into two hundred eighteen genera, assigned to sixty-five families according to Bentham and Hooker's system of classification have been recorded from Deedwana block. Halophytes are a unique group of plants that have adapted to thrive in high-salt environments such as coastal regions, salt marshes, or saline soils. These plants possess specialized mechanisms that enable them to tolerate and utilize the excess salt present in their surroundings. Halophytes play a crucial role in maintaining the ecological balance of saline habitats and contribute to biodiversity in these challenging environments. They have garnered significant scientific interest due to their exceptional ability to survive and grow under extreme conditions, which provides valuable insights into plant adaptation and stress tolerance mechanisms. Understanding the physiological and biochemical adaptations of halophytes can have implications for various fields such as agriculture, environmental conservation, and biotechnology. Phytochemical studies of halophytes are of great importance for several reasons. Firstly, halophytes are known to produce unique and diverse chemical compounds as a result of their adaptation to saline environments. These compounds, such as alkaloids, flavonoids, phenols, terpenoids, and others, have shown potential therapeutic properties, antioxidant activities, and ecological significance. Exploring the phytochemical composition of halophytes can provide valuable information about these bioactive compounds, which may have applications in traditional medicine, drug discovery, and natural product development. Secondly, phytochemical studies contribute to our understanding of the adaptive strategies employed by halophytes to survive in high-salt conditions. By elucidating the biochemical and physiological mechanisms underlying salt tolerance, researchers can identify key pathways and genes involved, which may have implications for improving crop plants' salt tolerance and addressing food security challenges in saline agricultural areas. Phytochemical investigations taken up for the present studies of some selected halophytic species viz Salsola baryosama, Suaeda fruticosa, and Trianthema triquetra occurring in Deedwana block of State Rajasthan include the quantitative estimation of total soluble sugars, soluble proteins, starch, free amino acids, phenols, and lipids. Representative halophyte species were selected based on their abundance, diversity, and ecological significance in the study area. Proper taxonomical identification of the collected species was done with the help of botanical experts to ensure accurate classification. A systematic collection of halophyte species was carried out in the study area, followed by sample preparation and extraction of phytochemicals. Material was obtained from various localities of Deedwana block.

Most of the areas have been reviewed by Jain (1970) Bhandari (1978), Sharma (1980). Publication of Flora of the Indian Desert (Bhandari, 1990), Flora of north-east Rajasthan (Sharma & Tiagi, 1979) and Flora of Rajasthan (Shetty & Singh, 1987) have further added to our knowledge of the flora and floral composition of Rajasthan. Quereishi (2002, 2017, 2018) and Sharma & Aggarwal (2008), have significantly contributed to our knowledge about the vegetation of Deedwana and Nagaur. In recent years a large number of publications dealing with the Halophytic flora and floral composition of Rajasthan have been published.  Vegetation Ecology of Halophytic Communities of Saline Arid studied by Hari and Dagar (2004). Contribution about halophytes of saline lands provided by Kumar (2015). Mangalassery Dayal and Patel, (2017b). Mohammed & Sen, (1987, 1988). Intensive botanical exploration of the Nagaur district of Rajasthan is in progress including a study of phytodiversity of Deedwana block. Some authors studied the Desert Plants in detail such as Ramawat (2010), Joshi, Bhanupriya, and Jaya Arora (2018).  Phytochemical analysis of some selected species of the family Convolvulaceae occurring in Central Rajasthan has been carried out recently by Sharma and Tomar (2023).

Material and Methods :

For the phytochemical study of halophytes, the three most common halophytic species viz. Salsola baryosma, Suaeda fruticosa, and Trianthema triquetra have been selected and Material for this purpose was obtained from plants growing at various localities of Deedwana block. Details of habitats, phenology, and soil characteristics were regularly recorded during the collection tours.

The methodology employed for this investigation is as follows:

Total Soluble Sugars: (Yem & Willis, 1964)

Reagents: 200 mg of Anthrone was dissolved in 100 ml ice cooled 70 percent sulphuric acid. the solution was stirred well. the fresh solution was used every time. 200 mg of dried material (leaves) was homogenized in 10 ml of 80 percent alcohol. After centrifugation the residue was again extracted with 10 ml of 80 percent alcohol and the two supernatants were combined and made up to a particular volume. an appropriate amount of aliquot and 4 ml of Anthrone reagent were mixed well and [placed in boiling water for 8 min. In blank supernatants were omitted. optical density was recorded at 800nm. A standard curve was prepared using glucose.

Total Soluble Proteins: (Lawry et al, 1951)

Reagents:

1. 02 percent Sodium carbonate (Na2CO3) in 0.1 N Sodium hydroxide (NaOH): 2 gm of Na2CO3 and 4 gm of NaOH dissolved in water and made up to 100 ml.

2. 0.3 percent Copper sulfate (CuSO4) in 1 % Na-K tartrate: 300 mg of CuSO4 and 1 gm of Na-K tartrate dissolved in distilled water made up to 100 ml.

3. Folin’s Reagent: Commercially available reagent used after two times dilution with distilled water.

Reagents (1) and (2) were mixed in a ratio of 50:1 shortly before use. A suitable quantity of dried material (200 gm) was homogenized in 10 ml of phosphate buffer (0.2 molar, pH=6.1). After centrifugation supernatants were used as a source of soluble proteins.

Total Starch:

Reagent: 0-2 % Iodine (I2) in 2 % KI. A suitable amount (200 gm) of dried material from each organ was homogenized in 10 ml of 80% ethanol (C2H5OH). After centrifugation, the residue was boiled in 10 ml of 1% KOH for 30 min. After centrifugation supernatant was used for the estimation of starch. To a suitable amount of aliquot was added 1 ml of reagent. In the blank, the aliquot was replaced by an equal amount of distilled water. Optical density was recorded after 10 min. at 600 nm. A standard curve was prepared using starch.

Total Phenols:

Reagents:

1. 30 % Sodium carbonate (Na2CO3) solution: 30 gm of Na2CO3 was dissolved in distilled water and made up to 100 ml

2. Folin’s Reagent: Commercially available reagent used after two times dilution with distilled water.

3. 200 mg homogenized in 10 ml. 80% alcohol. After centrifugation residue was again extracted with 10 ml of 80% alcohol. The supernatants were combined and made up to a specific volume and used as the source of total phenols.

The vegetation of Deedwana block in general, tolerates higher temperatures and intense solar radiation for most of the year. This includes the rainy season also, which is otherwise the most congenial period for the growth performances, of plants in natural habitats. The mean temperature during the rainy season is on an average 30⁰C during the day which is much higher compared to other parts of the Indian sub-continent, Ecological adaptations of the selected species of halophytes under hostile conditions of Deedwana block, Rajasthan has been investigated to evaluate the underlying eco-physiological characteristics of these plants:

A. Total Soluble Sugars (Table 1; Text Fig.1) :

Organwise, in root maximum sugar contents were in Salsola baryosma (17.9 mg/gdw), and minimum value was found in Sueada fruticosa (6.2 mg/gdw). and it was 11.4 mg/gdw in Trianthema triquetra. In stem the maximum sugar was recorded again in  Salsola baryosma (45.8 mg/gdw), followed by Sueada fruticosa (23.2 mg/gdw), the minimum being present in Trianthema triquetra (22.1 mg/gdw). In leaves, maximum sugar contents were observed in Salsola baryosma (40.7 mg/gdw), followed by Trianthema triquetra (30.9 mg/gdw), the minimum being present in Sueada fruticosa (28.6 mg/gdw). In general, the maximum amount of sugar contents was observed in leaves followed by stem, and the minimum was recorded in root of each of the investigated species.

B. Total Soluble Proteins ( Table 1; Text Fig.2) :

In the root, the maximum protein contents were found in Sueada fruticosa (8.1mg/gdw), followed by Salsola baryosma (5.6 mg/gdw), whereas Trianthema triquetra had a minimum amount of proteins (4.2 mg/gdw). In stem the maximum protein contents were found in Sueada fruticosa (84.6mg/gdw), followed by Trianthema triquetra (68.1 mg/gdw) with Salsola baryosma possessing a minimum amount  (47.4 mg/gdw). In leaves the maximum protein contents were found in Sueada fruticosa (68.6 mg/gdw), followed by Trianthema triquetra (51.8 mg/gdw), the minimum amount of proteins being in Salsola baryosma  (36.4 mg/gdw). Protein contents were comparatively maximum in stem and minimum in root with leaves an intermediate amount of the species except that the protein contents in the roots of Trianthema triquetra were less than that of Sueada fruticosa and Salsola baryosma

C. Total Starch (Table 1; Text fig. 3 ):

Organwise, in root maximum starch contents were observed in Trianthema triquetra (31.2 mg/gdw), followed by Salsola baryosma (13.8 mg/gdw). In stem maximum value was recorded in Trianthema triquetra (94.2 mg/gdw), the minimum being present in Salsola baryosma (41.3 mg/gdw). Similarly, the amount of starch was more in leaves of Trianthema triquetra (8.2 mg/gdw), as compared to Salsola baryosma (7.4 mg/gdw).In the inflorescence part the maximum starch contents observed in Trianthema triquetra (35.2 mg/gdw), followed by Salsola baryosma (17.5 mg/gdw) Generally, a very high quantity of starch was found in the stem of both these species whereas a minimum amount was found in the leaves and roots.

D. Total Free Amino Acids (Table 1; Text Fig. 4 ):

 Of the two species investigated for their amino acid contents Trianthema triquetra was found to have more of these acids as compared to Salsola baryosma. Organwise, In stem amino acid contents, were found to be more or less similar in both species viz., Trianthema triquetra (84.6 mg/gdw). and Salsola baryosma (76.4 mg/gdw). In leaves, the maximum amount of amino acid contents were recorded in Trianthema triquetra (94.7 mg/gdw) followed by  Salsola baryosma (84.8 mg/gdw). In the Inflorescence part, the maximum starch contents were observed in Trianthema triquetra  (81.6 mg/gdw).followed by  Salsola baryosma (75.2 mg/gdw).

Free amino acid contents in general were more in the stem followed by leaves and the minimum was recorded in the root in these species.

E. Total Phenols (Table 1; Text fig. 5 ):

Organwise, in the stem, maximum phenol contents were found in Trianthema triquetra (17.6 mg/gdw) and a minimum amount of phenols was observed in Salsola baryosma (13.8 mg/gdw). The amount of phenol in the leaves of all the species was almost similar to Trianthema triquetra (15.4 mg/gdw) and Salsola baryosma (15.2 mg/gdw). In the inflorescence part, Salsola baryosma (17.8 mg/gdw) had a maximum amount compared to Trianthema triquetra (12.9 mg/gdw).

F. Total Lipids ( Table 2; Text Fig.6) :

All three selected halophytic species occurring in the Deedwana block were studied for their lipid contents. Maximum lipid contents for the entire plant were recorded in Salsola baryosma (85.4 mg/gdw) and the minimum in Trianthema triquetra (34.9 mg/gdw) with Sueada fruticosa (65.2 mg/gdw) possessing value in between these two.

Total Polar and Non-Polar lipids ( Table 2; Text Fig. 6) :                                                             

Among the three species, maximum polar lipids were recorded in Salsola baryosma (22.6mg/gdw) followed by Suaeda fruticosa (21.1 mg/gdw) with Trianthema triquetra (10.9 mg/gdw) having minimum value. Among the investigated species, maximum non-polar lipid contents for the entire plant were recorded in Salsola baryosma (62.8mg/gdw)and the minimum in Trianthema triquetra (24.0mg/gdw) with Suaeda fruticosa (44.1 mg/gdw) possessing a value in between these two.

To a suitable amount of aliquot was added 3 ml of Na2CO3 solution and 0.5 ml Folin’s reagents. In the blank, the aliquot was replaced by equal volumes of distilled water. Test tubes were placed in boiling water for 1 minute and centrifuged to clear turbidity. Optical density was recorded at 800 nm.

Total Free Amino Acids:

Reagent:

1. Ninhydrin solution- 0.8 gm of reagent grade stannous chloride (SnCl2) dissolved in 500 ml of citrate buffer (pH=5). the solution is added to 500 ml of methyl cellosolve having 20 gm of Ninhydrin.

2. Diluent- A 50 % of iso-prapanol used as diluents.

A suitable amount of (200 mg) of dried material was homogenized in 10 ml of alcohol. After centrifugation residue was extracted with 10 ml of 80% alcohol. The two supernatants were combined. Chlorophyll pigments were removed by adding chloroform and water. the upper aqueous phases were used for the estimation of total free amino acids. To a suitable amount of aliquot was added 1 ml of Ninhydrin reagent. The test tubes were placed in boiling water for 20 min., immediately after this 5 ml of diluents was added and optical density was recorded at 570nm. A standard curve was prepared by using alanine.

Total Lipids:

Reagent:

500 gm of dried material was homogenized in Methanol: Chloroform mixture (2:1) and left at 40C overnight. It was filtered under a vacuum using sintered glass filter. The residue was repeatedly washed with Methanol. Chloroform mixture to ensure complete extraction of lipids. To this extract was added 0.9 % NaCl solution (one-third of the total volume) and left overnight. The lower phases were removed and dried in a vacuum at 50-550C. Total lipids were measured by gravimetric method.

Total polar and Non-polar lipids (Nichols)

Samples of total lipids were dissolved in 2-5 ml of Chloroform (CHCl3) and mixed with petroleum spirit: 95% of Methanol (1:1) and left overnight. The upper phase contains polar and the lower phase contains non-polar lipids. The two phases were separated and dried in a vaccum and weighed.

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