Int. J. Aquat. Biol. (2021) 9(4): 254-257 ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.ij-aquaticbiology.com © 2021 Iranian Society of Ichthyology Quality analysis and effect of reused water on some biological characteristics of Phaseolus vulgaris and Pisum sativum Batool M.H. AL-Adily *,1Noor M. Naji, Shaymaa O.H. Al-Mamoori Collage of Sciences, Babylon University, Hillah, Iraq. Article history: Received 7 April 2021 Accepted 21 August 2021 Available online 2 5 August 2021 Keywords: Reused water, P. vulgaris, P. sativum, Stem anatomy. Abstract: Seeds of Phaseolus vulgaris L. and Pisum sativum were planted in separated pots, then these pots divided into two groups, each group divided into four subgroups. Four different wastewater were used to irrigate the treatments. The results showed the effects induced by irrigated wastewater on P. vulgaris L. and P. sativum. The degree of water salinity induced increasing SOD, protein content and the area of xylem elements, but reduced chlorophyll-α in P. vulgaris. The CAT, protein and chlorophyll-α were increased in in P. sativum as a result to salinity which also reduces the area of xylem elements. Introduction Phaseolus vulgaris L. is a legume species with a wide distribution, therefore it can be grown in a high range of environmental conditions except those of extremes (Singh, 1999). Also, Pisum sativum is an edible leguminous seed that need a particular soil feature i.e. clay with slight acidity (Vural et al., 2000; Mani, 2015). In this species, salinity is more effective on germination of the bean (Ghezal et al., 2016). In Iraq, especially in past years, the regression of fresh water and increased the area with saline soil (Al-Adily, 2014) has been led to use other unusual water resources for irrigation (Al-Maamori and Al-Adily, 2018). Many works done regarding the effects of using wastewater for irrigation (Khurana and Singh, 2012; Nwaokobia et al., 2018). Since both P. vulgaris and P. sativum are sensitive to salinity (Maas and Hoffman, 1977; De Pascales et al., 1997; Munns and Tester, 2008), therefore, this work aimed to study the effects of different reused water for the irrigation of P. vulgaris and P. sativum as alternative for fresh water. Materials and Methods The seeds of P. vulgaris and P. sativum were planted in pots each with 1 kg soil, then seed pots of each *Correspondence: Batool M.H. AL-Adily DOI: https://doi.org/10.22034/ijab.v9i4.1347 E-mail: batool_aladily@yahoo.com species were divided into four subgroups each with three replicates. They were irrigated with four different wastewaters (Table 1). These wastewaters were collected from four different puncture water within Babylon Governorate, Iraq. Standard methods were used to measure of the water characteristics (APHA, 2005). In addition, the standers methods of soil labs were used to measure the important features of the used soil in the palnted pots (ICARDA, 2001). The measure characteristics are shown in Table 1. The biochemical measurement of the plants with ages of 45 day were chlorophyll-α (Yash, 1998), proline content (Bates, 1973), SOD (Marklund and Marklund, 1974), total protein (Bradford, 1976) and catalase content (Aeibi, 1984). The method to study the anatomical variations was based on Jahanson (1940). Results and Discussions The biochemical results of P. vulgaris L. and P. sativum showed their different strategies to adapt with water quality (Tables 1, 2). The high salinity of type 3 water (EC=3320 μs/cm) reduce chlorophyll-α and CAT in P. vulgaris, which may be due to concentration of chloride because bean is sensitive to salinity ions (Bouzid and Rahmoune, 2012; Anna et 255 Int. J. Aquat. Biol. (2021) 9(4): 254-257 al., 2015) causing defect in amount of gas exchange and the producig chlorophyll-α (Kauymakanova and Stoeva, 2008). Whereas, P. sativum do not showed an increase in chlorophyll-α content with increase salinity of irrigated water and high concentration recorded in group 4 containing high concentration of organic matter (0.28%). Pisum was able to tolerant salinity by increasing proline formation and protein content which was in a positive relationship with CAT. This results are in agreement with other studies done on pisum on other type of soil and under different weather conditions (Mani, 2015; Artega et al., 2020). The results also give a good indicator to prove ability of pisum agriculture in Iraq. Histological sections of the stem in the studied species (Figs. 1 and 2) revealed that thickness of epidermal cells was higher in group 2 which irrigated with wastewater containing lower concentrations of total ions and salinity, while the high levels of salinity induced increasing of the epidermal layer numbers and no increase found in group 3, so in this group showed an increasing in area of xylem elements and the higher increase was in group 4. Bean is adapted to salinity in many different adaptation, some were Table 1. Some characters of four types of wastewater used in irrigated of Phaseolus vulgaris and Pisum sativum. Wastewater Parameters 4 3 2 1 8.3 8.5 8.1 8.6 pH 3210 3320 950 1438 E.C (μs/cm) 1450 2340 698 1030 TDS (mg/l) 1740 2930 786 1240 Salinity (mg/l) 920 1480 840 700 Total Hardness (mg/l) 368.74 593.18 336.67 280.56 Calcium (mg/l) 134.51 216.38 122.81 102.34 Magnesium (mg/l) 499.85 779.76 409.76 349.89 Chloride (mg/l) 320 400 240 140 Alkalinity (mg/l) 241.18 153.92 122.55 67.65 Sulphate (mg/l) 0.28 0.12 0.16 0.07 Organic matter (%) Figure 1. Anatomical variation of stem of Phaseolus vulgaris induced by different wastewater sources (20X). 256 AL-Adily et al./ effect of reused water on some biological characteristics of P. vulgaris and P. sativum biochemically or phonologically (Al-Hassan et al., 2016) and in this study anatomical adaptations, including decrease the layers of collenchyma tissue was observed. In contrast, in pisum, there was no significant variations in the area of epidermis, while organic matter may be induced the producing of the parenchyma tissue and the width of vascular bands (as seen in group 4) and the number of the xylem elements but it reduced the diameter of xylem vessel. Plants can tolerant elevated concentrations of salinity ions by increasing the fibers of xylem (Al-Adily et al., 2018). Salinity causes some plants to change the anatomy of vascular system according to degree of EC of water or soil (Tietjen et al., 2017; Qaderi et al., 2019). References Aeibi H. (1984). Catalase in vitro. Methods of Enzymology, 105: 121-126. Al-Hassan M., Morosan M., Pilar M., Prohens J., Vicente O., Boscauo M. (2016). Salinity-Induced Variation in Biochemical Markers Provides Insight into the Mechanisms of Salt Tolerance in Common (Phaseolus vulgaris) and Runner (P. coccineus) Beans. International Journal of Molecular Sciences, 17(9): 1582. Table 2. Physiological responses of Phaseolus vulgaris and Pisum sativum to type of irrigated waste water. Chlorophyll content (SPAD) proline (µmole / g.D.W) protein (mg/ g.D.W) SOD (U/ml) CAT (U/ml) Wastewater Species 25.4 16.85 23.74 4.477 22.24 1 P. vulgaris 22.9 20.48 24.79 2.947 15.09 2 12.4 22.22 27.8 3.91 11.32 3 29.5 23.15 26.96 3.343 30.48 4 17.3 48.63 24.4 2.493 13.81 1 P. sativum 21.7 47.82 27.8 1.45 16.02 2 23.4 59.55 27.96 1.813 18.11 3 29.3 60.79 24 2.55 15.07 4 Figure 2. Anatomical variation of stem of Pisum sativum induced by different wastewater sources (20X). 257 Int. J. Aquat. Biol. (2021) 9(4): 254-257 Al-Adily B.M., Al-Ani H.A., Al-Taee M.M. (2016). Anatomical variation of some halophytes from different sites in Babylon Provinus, Iraq. Mesopotamia Environmental Journal, 2(2). Al-Adily B.M.H. (2014). Study of some halophytic vegetation environments in Babylon Governorate/Iraq. A thesis, college of Science–University of Babylon. Al-Mamoori S.O., Naji N.M., Sura I.A.J., Kamil Z.S., Naji O.M. (2020). Efficiency of Vigna radiata and Cucumis sativus for the accumulation of heavy elements in their tissues. Plant Archives, 20(Supplement 1): 2824-2826. Anna S., Salmas I., Nifakos K., Kalogeropolous P. (2015). Effect of Salt stress on three green bean (Phaseolus vulgaris L.) cultivars. Not Bot Horti Agrobo, 43(1): 113-118. APHA (American public health association). (2005). Standard methods for examination of water and wastewater.21the Ed. Washington, D.C., U.S.A. Artega S., Yabor L., Diez M., Prohens J., Boscaiu M., Vicente A. (2020). The Use of Proline in Screening for Tolerance toDrought and Salinity in Common Bean (Phaseolus vulgaris L.) Genotypes. Agronomy, 10: 817. Bates L.S., Waldren R., Teare I.D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil, 39: 205-207. Bouzaid S., Rahmoune, C. (2012). Enhancement of Saline Water for Irrigation of Phaseolus vulgaris L. Species in Presence of Molybdenum. Procedia Engineering, 33: 168-173. Bradford M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2): 248-254. De Pascale S., Barbieri G., Ruggiero C. (1996). Effects of water salinity on plant growth and water relations in snap bean (Phaseolus vulgaris L.). In II International Symposium on Irrigation of Horticultural Crops, 449: 649-656). Ghezal N., Rinez I., Sbai H., Farooq M., Rinez A., Zribi I., Haouala R. (2016). Improvement of Pisum sativum salt stress tolerance by bio-priming their seeds using Typha angustifolia leaves aqueous extract. South African Journal of Botany, 105: 240-250. ICARDA. (2001). Organic matter determination. International Center for Agricultural Research in drier areas, 2ed. Johanson A.D. (1940). Plant microtechnique. 1st (Eds.) Mc. Graw. Hill Book Company, New York and London, 523 p. Kaymakanova M., Stoeva N. (2008). Physiological reactions of bean plants P.vulgaris to salt stress, Gen. Appl. Plant Physiology, 34(3-4): 177-188. Khurana M.P.S., Singh P. (2012). Waste water use in crop production: a review. Resources and Environment, 2(4): 116-131. Maas E.V., Hoffman G.J. (1977). Crop salt tolerance- current assessment. Journal of the Irrigation and Drainage Division, 103(2): 115-134. Mani F. (2015). Effect of salt stress on physiological attributes of pea (Pisum sativum). International Journal of Agricultural Science and Research (IJASR), 5(1): 29- 41. Marklund S., Marklund G. (1974). Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry, 47(3): 469-474. Munns R., Tester M. (2008). Mechanisms of salinity tolerance. Anneal Review of Plant Biology, 59: 651- 681. Nwaokobia K., Ogboru R.O., Idibie C.A. (2018). Effects of grey water irrigation on the cultivation of African spinach (Amaranthus hybridus). World News of Natural Sciences 18(2): 133-145. QadEri M.M., Martel A.B., Dixon S.L. (2019). Environmental Factors Influence Plant Vascular System and Water Regulation. Plants, 8: 65. Qasim M.T., Al-Mayali H.K. (2019). Investigate the relation between Baicalin effect and gene expression of LH, FSH, Testosterone in male rats treated with Gemcitabine drug. Research Journal of Pharmacy and Technology, 12(9): 4135-4141. Qasim M.T., Al-Mayali, H.K. (2019). The immunological and protective role of baicalin in male rats treated with chemotherapy (Gemcitabine). Journal of Physics Conference Series, 1234:012065. Singh, S.P. (1999). Developments in plant breeding: Common bean improvement in the twenty-first century. Kluwer Academic Publishers, the Netherlands. 409 p. Tietjen B., Schlaepfer D.R., Bradford J.B., Lauenroth W.K., Hall S.A., Duniway M.C., Hochstrasser T., Jia G., Munson S.M., Pyke D.A. (2017). Climate change- induced vegetation shifts lead to more ecological droughts despite projected rainfall increases in many global temperate dry lands. Global Change Biology, 23(7): 2743-2754. Wilkinson S., Davies W.J. (2008). Manipulation of the apoplastic pH of intact plants mimics stomatal and growth responses to water availability and microclimatic variation. Journal of Experimental Botany, 59(3): 619-631. Yash K.P. (1998). Handbook of references methods for plant analysis. Soil and Plant Analysis Council, Inc. CRC Press. Taylor and Francis Group. 291 p.