Ameliorating Role Of N.P.K Fertilizer On The Toxic Effects Of Ni On (Sorghum) Root Antioxidant Enzymes

Project and Seminar Material for Biochemistry

Ameliorating Role Of N.P.K Fertilizer On The Toxic Effects Of Ni On (Sorghum) Root Antioxidant Enzymes


Abstract


This study investigated the activities of superoxide dismutase (SOD), catalase (CAT), glutothione peroxidase (GP) and the level of malondialdehyde (MDA) in the root of sorghum grown in soils contaminated with 30ppm nickel, 30ppm nickel +20ppm fertilizer and 30ppm nickel + 40ppm fertilizer. Sixty sorghum seeds were germinated in these contaminated soils and were harvested after 2 weeks, 3 weeks, and 4 weeks of planting. Treatment of the plants with 30ppm nickel significantly increased (P < 0.05) the activities of SOD and the level of MDA in the roots compared with the controls. Also, the treatment significantly decreased (P < 0.05) the activities of CAT and GP in the roots compared with controls.

The study also revealed a significant decrease (P < 0.05) in the activities of SOD and the level of MDA in plants grown in 30ppm Ni + 20ppm NPK fertilizer and 30ppm Ni + 40ppm NPK fertilizer respectively compared with those grown in 30ppm Ni concentration. These results show that 30ppm Nickel is toxic to sorghum roots for it increases significantly the production of reactive oxygen species but decreases significantly the excretion of reactive oxygen species. This is due to significant increase in the activity of SOD but significant decrease in the activities of CAT and GP. These results also showed that 30ppm Nickel damaged sorghum roots by significantly increasing lipid peroxidation and the levels of MDA. In addition, the results revealed that 20ppm and 40ppm NPK fertilizer had ameliorating effect on the toxicity caused by 30ppm nickel.


Table Of Contents


  • Title Page i
  • Certification ii
  • Dedication iii
  • Acknowledgement iv
  • Table of Contents vi
  • Abstract ix

Chapter One

  • Introduction and Literature Review 1
  • 1.1 Introduction 1
  • 1.2 Literature Review 3
  • 1.2.1 Definition of heavy metals 3
  • 1.2.2 Characteristics of Nickel 4
  • 1.2.3 Nickel in the environment 5
  • 1.2.4 Biological roles of nickel 6
  • 1.2.5 Absorption of nickel by plant 7
  • 1.2.6 Accumulation of Nickel in plants 9
  • 1.2.7 Nickel and photosynthesis 10
  • 1.2.8 Effects of nickel on plant respiration 10
  • 1.2.9 Metabolic effects of nickel 11
  • 1.2.10 Effects of nickel on enzyme activity 11
  • 1.2.11 Mechanism of nickel toxicity 12
  • 1.2.12 Strategies of plant tolerance to nickel toxicity 14
  • 1.2.13 Management of nickel toxicity 18
  • 1.2.13.1 Land management procedure 18
  • 1.2.13.2 Phytoremediation 19
  • 1.2.14 The use of micro-organisms to mitigate nickel toxicity 22
  • 1.3 Scientific classification of Sorghum 23
  • 1.4 Chemical Composition and Nutritive Value of Sorghum 25
  • 1.5 Classification of sorghum 28
  • 1.6 Uses of Sorghum 29
  • 1.7 Germination / Growth Stages of Sorghum 32
  • 1.7.1 Growth Stages 32
  • 1.72 Nutrient Uptake 36
  • 1.8 Diseases of Grain Sorghum 37
  • 1.9 Activities that induce Germination 38
  • 1.10 Metabolism of Germinating Seeds 40
  • 1.11 NPK (15-15-15) Fertilizer 41
  • 1.11.1 Catalase 44
  • 1.12 History of Catalase 45
  • 1.12.1 Activities of Catalase 46
  • 1.12.2 Molecular mechanism of catalase action 47
  • 1.13 Superoxide Dismatase 48
  • 1.13.1 Types of Superoxide Dismutase 49
  • 1.13.2 Physiological Importance of Superoxide Dismutase 51
  • 1.13.3 Use of Superoxide Dismutase in Cosmetic 52
  • 1.14 Peroxidase 52
  • 1.14.1 Isozymes of Glutathione Peroxidase 53
  • 1.15 Oxidative stress and reactive oxygen species 53
  • 1.16 Objective of the Study 56

Chapter Two

  • Materials and Method 55
  • 2.1 Materials 55
  • 2.1.1 Contaminant 55
  • 2.1.2 Fertilizer 55
  • 2.1.3 Quantity of soil used 55
  • 2.1.4 Source of Soil 55
  • 2.1.5 Source of Soybean seed used 56
  • 2.1.6 Instruments/Apparatus used 56
  • 2.1.7 Reagents used for the study 57
  • 2.1.8 Methods 59
  • 2.1.9 Preparation of Soil 59
  • 2.1.10 Contamination of Soil 59
  • 2.1.11 Viability test of Seeds 59
  • 2.1.12 Experimental design 59
  • 2.2 Biochemical analysis 62
  • 2.2.1 Estimation of total protein 61
  • 2.2.2 Estimation of malondialdehyde level 64
  • 2.2.3 Estimation of Superoxide Dismutase activity 66
  • 2.2.4 Estimation of Catalase activity 68
  • 2.2.5 Estimation of peroxidase activity 70
  • 2.2.6 Statistical Analysis 72

Chapter Three

  • Results 73
  • 3.1 Soil Analysis 78

Chapter Four

  • Discussion and Conclusion 79
  • Bibliography 83
  • Appendix One; Reagents Preparation 97
  • Appendix Two; Statistics 101

Chapter One


Introduction And Literature Review

1.1 Introduction

Trace metals are redistributed in environment by fossil fuel combustion. This release can be expected to increase soil levels of trace elements such as Ni2+ resulting in a concomitant increase in the concentration of Ni2+ in plants and possibly in the food chain (Dominic et al, 1978).

Nickel (Ni) is an essential micronutrient for plants since it is the active centre of the enzyme urease required for nitrogen metabolism in higher plants (Yan et al, 2008). Nickel deficiencies lead to reduced urease activity in tissue cultures of sorghum, rice and tobacco and in excessive accumulation of urea and toxic damage to the leaves of leguminous plants such as sorghum (Peter and Andre, 1986). However, excess Ni is known to be toxic and many studies have been conducted concerning Ni toxicity of various plant species.

The most common symptoms of nickel toxicity in plants are inhibition of growth, photosynthesis, mineral nutrition, sugar transport and water relations (Seregin and Kozhevnikova, 2006). Heavy metal affects plants in two ways. First, it alters reaction rates and influences the kinetic properties of enzymes leading to changes in plant metabolism (Yan et al, 2008). Second, excessive heavy metals lead to oxidant stress.

During the period of metal treatment, plants develop different resistance mechanisms to avoid or tolerate metal stress, including the changes of lipid composition, enzyme activity, sugar or amino acid contents, and the level of soluble proteins and gene expressions. These adaptations entail qualitative and/or quantitative advantage, and affect plant existence (Schutzendubel and Polle, 2002).

It is known that excessive heavy metal exposure may increase the generation of reactive oxygen species (ROS) in plants, and oxidative stress would arise if the balance between ROS generation and removal were broken. Oxidative stress is a part of general stress that arises when an organism experiences different external or internal factors changing its homeostasis. In response, an organism either aims to maintain the previous status by activation of corresponding protective mechanisms or goes to a new stable state (Mittler, 2002).

In several plants, Ni has been shown to induce changes in the activity of ROS – scavenging enzymes, including SOD catalase and glutathione peroxidase (Yan et al, 2008).

The aim of this study is to investigate the effects of nickel on the activities of sorghum root antioxidant enzymes and also monitor the ameliorating effects of N.P.K. Fertilizer.


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