BIO-FORTIFICATION OF Murraya koenigii and Vernonia amygdalina WITH IODINE

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ABSTRACT
This study was aimed at increasing the level of iodine in commonly consumed vegetables by fortifying them and help reach the important health and social objective of IDD elimination. Two plant species(Murraya koenigii and Vernonia amygdalina) were cultivated, fortified and analysed for concentration of iodine uptake. The seedlings of these plants were obtained from Eke-Awka market, cultivated and were inoculated with different concentrations of potassium iodide and potassium iodate, namely 0.1M, 0.2M, 0.3M and 0.4M of both KI and KIO3. After two weeks, the leaves of these plants were taken for iodine analysis using the alkaline dry ash technique. the result shows that in Murraya koenigii, the various concentrations of 0.05M, 0. 1M, 0.2M and 0.3M KI gave iodine values of 26.014, 0, 0, 12.056,. concentrations of 0.1M, 0.2M, 0.3M and 0.4M KIO3 gave iodine values of 1.269, 0, 16.497, 26.0 15. In Vernonia amygdalina, the various concentrations of 0.05M, 0.1M, 0.2M and 0.3M KI gave iodine values of 1.904, 22.842, 16.497, 27.284. concentrations of 0.1M, 0.2M, 0.3M and 0.4M KIO3 gave iodine values of 0, 0, 14.59, 0. The results also show that plants were able to tolerate different levels of iodine concentrations as KIO3 better than KI in the root environment. Between the two cultivated species, Murraya koenigii showed greater tendency of iodine accumulation than Vernonia amygdalina.

TABLE OF CONTENTS
Title page — — — — — — — — — i
Certification– — — – — — — — — ii
Dedication — — – — — — — — — iii
Acknowledgement — — — — — — — — iv
Table of Contents — — — — — — — — v
List of Tables — — — — — — — — x
Abstract — — — — — — — — — xi
CHAPTER ONE
1.0 Introduction — — — — — — — — 1
1.1 Importance of the study — — — — — — 4
1.2 Significance of the study — — — — — — 5
CHAPTER TWO
2.0 Literature Review — — — — — – — 6
2.1 Bitter Leaf (Vernonia amygdalina) — — — — 6
2.1.1 Ecology — — — — — — — 6
2.1.2 Major Uses and Functions — — — — 7
2.1.3 Feeding Value — — — — — — 7
2.1.4 Scientific Classification — — — — — 8
2.1.5 Chemical Composition Of Vernonia amygdalina Collected
From Different Reference — — — — — 9
2.2 Curry (Murraya koenigii) — — — — — — 13
2.2.1 Scientific Classification — — — — — — 13
2.2.2 Description — — — — — — — — 14
2.2.3 Uses — — — — — — — — — 14
2.2.4 Chemical Constituents — — — – — — 14
2.2.5 Identification Test — — — — — — — 15
2.3 Iodine– — — — — — — — — 15
2.3.1 Where Is Iodine Found In The Body? — — — — 16
2.4 Iodine Deficiency Diseases (IDDs) — — — — 18
2.4.1 WhatIsIDD? — — — — — — — 18
2.4.2 What Are The Signs And Symptoms Of Iodine Deficiency? 19
2.4.1 What Causes IDD? — — — — — — 21
2.4.2 How Is IDD Measured? — — — — — — 23

CHAPTER THREE
3.0 Materials and Methods — — — — — — 28
3.1 Equipment Used — — — — — — — 28
3.2 Reagents — — — — — — — — 28
3.3 Preparation of Reagents — — — — — — 29
3.4 Cultivation of Plants — — — — — — — 36
3.5 Treatment of Plants — — — — — — — 36
3.6 Harvest — — – — — — — — — 37
3.7 Determination of Iodine Value of oil (Wij’s Method) — — 37
CHAPTER FOUR
4.0 Results and Discussion — — — — — — 39
4.1 Results — — — — — — — — — 39
4.2 Calculations — — — — — — — — 41
4.3 Discussion — — — – — — — — 44
Conclusion — — — — — — — — — 47
References — — — — — — — — — 48

LIST OF TABLES
Table 1: Basic Information about iodine — — — — 2
Table 2. Chemical Composition Of Vernonia amygdalina — — 12
Table 4.1.0 showing the result of Iodine value of KI concentrations in bitter leaf — — — — — — — 39
Table 4.1.1 showing the result of Iodine value of KIO3 concentrations in bitter leaf — — — — – — — 39
Table 4.1.3 showing the result of Iodine value of KI concentrations in curry — — — — — — — — 40
Table 4.1.4 showing the result of Iodine value of KIO3 concentrations in Curry — — — — — — — — 40
ix
1
CHAPTER 1
1.0 Introduction
Iodine is a trace mineral and an essential nutrient found naturally in the human body. It is needed for the metabolism of cells, for normal thyroid function and for the production of thyroid hormones.
1.1 Food Sources of Iodine
Include iodized salt which is the main food source of iodine. Sea food is also naturally rich in iodine, kelp is the most common vegetable seafood that is a rich source of iodine, and dairy products also contain iodine. Other good sources are plants grown in iodine – rich soil.
Lack of enough iodine(deficiency) may occur in places that have iodine- poor soil. Many months of iodine deficiency in a person’s diet may cause goiter or hypothyroidism. Without enough iodine the thyroid cells and the thyroid gland become enlarged.

Deficiency happens more often in women than in men, and it is more common in pregnant women and older children. Getting enough iodine in the diet may prevent a form of physical and mental retardation called cretinism.
Table 1: Basic Information about iodine.
Name Iodine
Symbol I
Atomic number 53
Atomic weight 126.90447
Standard state Solid at 298k
Group in periodic table 17
Group name Halogen
Period in periodic table 5
Block in periodic table P-block
Colour Violet-dark grey, lustrous
Classification Non-metallic

In the last few years numerous studies have been carried out on iodine bio fortification (enrichment) of plants [white and broadly 2005, 2009; Yang et al., 2007; Zhao and McGrath, 2009].

Studies presented in this work respond to the goals proposed by the WHO program. Iodine is not a mineral nutrient essential for plant growth and development. For that reason, evaluation of agronomic rules of its application requires thorough research focus not only on optimization of bio fortification but also on iodine effect on the quantity and quality of yield. This evaluation is so important as iodine, most probably, affect nitrogen metabolism in plants which contributes the most to plant productivity. This assumption is indirectly confirmed by studies conducted by Tsugonai andSase [1969] on Escherichia Coli,
Extracts as well as Oy Wong and hung [2001] and hung et al. [2005] on marine phytoplankton.
Iodine bio fortification of plants through soil fertilization is relatively low effective, which is caused by strong iodine sorption in soil. Plant uptake of iodine depends on its availability, which is initially governed by the adsorption – desorption. Characteristics soils [Dai et al. 2009]. Three days after introduction to soil, approximately 90% of iodine is strongly bound by aluminum and iron sesquioxides [muramatsu et al. 1990, Yoshida da et al.

1992]. The process of desorption is very slow that results in low level of iodine in soil solution and, furthermore, limits its uptake by plants [Quge and John sun, 1986, muramatsu et al. 1996; Yamaguchi et al. 1996; Yamaguchi et al.2005].
All attempts of enriching plants with iodine by fertilization with high doses bring risks of plant damage due to toxic effects of excessive iodine levels (Smith and Middleton 1982; Mackowiak and Gross, 1999; Mackowiak et al. 2005; Hong et al.2009). A significant issue undertaken in studies on iodine bio fortification is how to increase soil level of iodine forms available for plants without applying too high iodine dose. Works presented by Muramatsu et al. (1996) proposed the possible solution of enhancing iodine desorption from soil which is observed with negative values of soil redox potential (EH). Most traditionally cultivated soils are characterized by positive Eh and negative values of this parameter are noted after prolonged flooding which induces anaerobic conditions in soils [Astbury, et al., 1999].

1.2 Aim of the Study
The aim or objectives of the study was to determine the effect of iodine form
and the method of iodine fertilization on iodine bio fortification. [Astbury, et
al., 1999].
The use of salt has its attendant problems which include: recurrent cost of production, delivery network, storage ability and monitoring (Babikir, 1994) and some health implications (Feid. Ramussen, 2001; Delange et al., 1999; Laurberg et al.,). In such situation, alternative methods of IDD prevention such as iodization of water used for the irrigation of plants may be necessary for some category of people who may not benefit from iodized salt either for health or other reasons. We may therefore say that salt fortification with iodine is not enough to eliminate IDDs which are primarily the result of inadequate amounts of iodine in soil, water and food, resupplying iodine to the soil medium surely increases the iodine available for plant uptake. Therefore the objectives of this study were to develops a method of improving the iodine content of these two leafy vegetable plants: Vernonia

amygdalina (commonly known as bitter leaf) and Murraya koenigii (curry
leaf) by the application of weighed amounts of potassium iodide (KI) and
potassium iodate (KIO3) to the soil or pre-sowing KI and KIO3fertilization
and examining the effects of the applied iodate on the vegetable growth by determining the proximate compositions.
1.3 Importance of the Study
Excessive consumption of table salt is in many countries one of the main contributors to increased occurrences of cardiovascular diseases. For that reason, WHO has developed “The Global strategy on Diet, physical Activities Health” on years 2008-2013. This includes limitation of salt consumption with concurrent search for effective search for effective ways of introducing iodine into food chain, which is crucial due to the numerous functions played by this element in human organism [Astbury, et al., 1999].

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