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EFFECTS OF CHEMICAL MUTAGENS ON PHENOTYPIC AND CRYOGENIC VARIABILITY IN GARLIC

ABSTRACT

The study investigates the influence of chemical mutagens on the phenotypic traits and cryogenic variability of garlic (Allium sativum L.), a critical crop with significant agricultural and culinary importance. Mutagenesis, a process of inducing mutations to enhance genetic diversity, has been applied using chemical agents such as ethyl methanesulfonate (EMS) and sodium azide (SA). The primary objective is to assess the resultant phenotypic changes, including alterations in growth patterns, bulb morphology, and resistance to biotic and abiotic stresses. Additionally, the study evaluates cryogenic variability by examining the genetic stability and viability of garlic genotypes post cryopreservation.

Field trials and controlled environment experiments were conducted to compare treated and untreated garlic populations. Detailed phenotypic assessments revealed significant variations in traits such as plant height, leaf length, bulb size, and clove number, indicating the mutagenic potential of EMS and SA. Molecular analysis through techniques like RAPD and AFLP provided insights into genetic polymorphisms induced by the mutagens. Cryopreservation studies involved evaluating garlic samples stored in liquid nitrogen, with subsequent analysis of their survival rate, regrowth potential, and genetic integrity.

Table of Contents

Chapter 1: Introduction

1.1 Background and Rationale

1.2 Objectives of the Study

1.3 Hypotheses

1.4 Significance of the Study

1.5 Structure of the Thesis

Chapter 2: Literature Review

2.1 Garlic: Botanical Characteristics and Economic Importance

2.2 Genetic Variability and Breeding in Garlic

2.3 Chemical Mutagenesis: Mechanisms and Applications

2.4 Phenotypic Variability: Assessing Plant Traits

2.5 Cryogenic Techniques in Plant Science

2.6 Previous Studies on Mutagenesis and Cryopreservation in Garlic

2.7 Knowledge Gaps and Research Needs

Chapter 3: Materials and Methods

3.1 Plant Material

3.2 Experimental Design

3.2.1 Field Trials

3.2.2 Controlled Environment Experiments

3.3 Chemical Mutagen Treatment

3.3.1 Selection of Mutagens (EMS, Sodium Azide)

3.3.2 Treatment Protocols

3.4 Phenotypic Assessment

3.4.1 Growth Parameters

3.4.2 Morphological Traits

3.4.3 Stress Resistance

3.5 Molecular Analysis

3.5.1 DNA Extraction

3.5.2 RAPD and AFLP Techniques

3.6 Cryogenic Preservation

3.6.1 Sample Preparation

3.6.2 Cryopreservation Protocols

3.6.3 Post-Thaw Viability and Regrowth Assessment

3.7 Data Analysis

3.7.1 Statistical Methods

3.7.2 Interpretation of Results

Chapter 4: Results and Discussion

4.1 Phenotypic Variability Induced by Chemical Mutagens

4.1.1 Growth Patterns and Leaf Traits

4.1.2 Bulb Morphology and Clove Characteristics

4.1.3 Stress Resistance Traits

4.2 Molecular Analysis of Mutagenic Effects

4.2.1 Genetic Polymorphism via RAPD

4.2.2 AFLP Analysis of Genetic Variability

4.3 Cryogenic Variability and Genetic Stability

4.3.1 Survival Rates Post-Cryopreservation

4.3.2 Regrowth Potential and Morphological Integrity

4.3.3 Molecular Integrity After Cryopreservation

4.4 Comparative Analysis with Untreated Controls

4.5 Implications for Garlic Breeding Programs

4.6 Discussion of Results in Context of Existing Literature

Chapter 5: Conclusion and Recommendations

5.1 Summary of Findings

5.2 Conclusions

5.3 Practical Implications

5.4 Recommendations for Future Research

5.5 Final Thoughts

References

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