1. Experiment No 2

2. 5 Experiment Material and Chemicals Overview Introduction Procedure Objective 1 2 3 4

3. Introduction Introduction

4. Water containing dissolved substances such as chlorides, sulfates, carbonates, and bicarbonates of calcium, magnesium, sodium, and potassium in high concentration is called as high salt water. Low concentration of dissolved salt will become cause of low salt water. High and low salt water

5. Threat of squandering of one third of cultivated land by desertification. Limited sweet water resources and increase in the use of brackish underground water,as the main reasons of desertification in Pakistan. Abiotic stresses for crops Drought light Heat salt stress Effect of salt concentration on crops

6. Excess salinity in soil water can decrease plant available water and cause plant stress. Salt tolerance of plant depends upon its ability to check the movement of salts at five sites. Selective absorption of roots. Entry of nutrients in xylem. Entry of food materials in the phloem. Exclusion of salts. Evacuation of toxic salts by salt glands. Effect of high salt water: Wilting of plant

7. High or low salt concentration become cause of fluctuation in chloride and sodium ions of different parts of plant. Different critical mechanisms of plant Photosynthesis Stomatal conductance Transpiration rate Effects

8. Salt tolerance and salt sensitivity Lower conductivity of distilled water than tap water Types and Causes of Salinity human-induced salinity Example of salinity

9. Experiment for In vitro Seed Germination Experiment for In vitro Seed Germination

10. Filter paper Tap water NaCl Distilled water Petri dishes Agar HgCl 2 Tissue paper Balance Cylinder Beaker Apparatus and Chemicals

11. Layout and protocol of the Experiment Layout and protocol of the Experiment

12. Preparation of Agar Tap,distilled and salted water is used Seed Preparation Soaking of seeds Agar +Tap water Autoclave to avoid contamination Surface Sterilization of seeds 70% ethanol is used Laminar flow hood is used Do not dip into ethanol overnight

13. Washing of seeds Seed coat is removed Propagation of seed germination Transfer of seeds in test Petri dishes Agar+ prepared seeds or Filter paper +prepared seeds Provide favorable conditions for growth Placement in growth room And observation of results Use of Laminar flow hood+ hands should be washed properly

14. Soak seeds in distilled water, tap water and salted water overnight. In order to prepare plain agar dissolve 1 gram of agar is in 100 ml of tap water and autoclave it with other necessary apparatus in autoclave at 120c for 20 min. Take all apparatus in laminar flow to avoid contamination. For surface sterilization of seeds dip them into ethanol solution for 2-3 minutes. Protocol

15. Wash them 2-3 times and then remove seed coats. Place those soaked and rest of the un soaked seeds in Petri dish with the help of forceps, in which solidified agar media is present. Place filter paper in Petri dish and pour 5 ml distilled water on it and then place soaked and un soaked seeds on it for germination. Clover Petri plates with aluminum foil and place them in growth room for a day and check the growth of roots and shoots. Record observation in form of table daily for one week.

16. Seeds will germinate more quickly if they are first placed in a cup of tap water and allowed to sit and soak-up water. Do not dip in HgCl2 overnight. Avoid contamination during experiment. Agar should be fully dissolved in water for media preparation. Observation Precautions

17. Thank You