Jackfruit (Artocarpus heterophyllus) is commonly grown in home gardens of tropical and sub-tropical countries. The fruit contains high levels of carbohydrates, protein, starch, calcium and vitamins. Jack fruit has diverse medicinal uses, especially anti-oxidant, anti-inflammatory, anti-microbial, anti-cancer and anti-fungal activity. Jackfruit is considered to be an underutilized fruit where most of the fruits get wasted due to ignorance, lack of post harvest technology and gaps in supply chain systems.
Jackfruit contains more protein, calcium, iron, vitamins and other essential nutrients as compared to the common fruits. To find out the morphological variation among jack fruits among Kerala we conducted an elaborative survey and found out the parameters such as tree characteristics, leaf characterises or fruit characteristics of selected samples.
Both qualitative and quantitative data are measured using field visits and standard methods and morphological indicators; after analysis of data there is a morphological variation among the jack fruit samples identified. Advances in the genetic markers such as RFLP and PCR based methods are more reliable for identification of genetic diversity than morphological markers although each technique has advantages and limitations.
The objective of this research work was to estimate the level of genetic diversity and to assess genetic relationships among six varieties of jackfruit using the ‘rbcL gene’ based on PCR technique and RFLP markers. The partial sequence of ‘rbcL’ gene of six different Artocarpus varieties was used in the analysis. The size of amplified products was approximately 700 bp. After sequencing and sequence editing, sequence information on a 651 bp region was finally obtained for analysis. The alignment of sequences revealed two haplotypes out of 651 sites. The nucleotide frequencies are 27.96% (A), 29.47% (T/U), 19.69% (C), and 22.89% (G). Being one of the underutilized fruits in India, Artocarpus heterophyllus Lam has promising leads to further scientific research and livelihood strategies. The study of rbcL gene using PCR and RFLP seems to be a promising tool in establishing genetic diversity among jackfruit varieties.
Table of Contents
1. Introduction
1.1Taxonomical classification
2. Review of literature
3. Hypothesis
4. Materials and Methods
4.1 Botanical description and varieties
4.2 Nutritional and vitamin composition
4.3 Study area
4.4 Sample collection
4.5 Morphological descriptors for jackfruit tree classification
4.6 Isolation of DNA
4.7 Quantification of DNA
4.8 PCR amplification
4.9 Data sequencing
4.10 Data analysis
4.11 Statistical analysis
5. Results and discussion
5.1 General characteristics
5.2 Leaf characteristics
5.3 Fruit
5.4 DNA isolation
5.5 Polymerase chain reaction
5.6 Molecular phylogentic analysis using ‘rbcL’ gene
5.7 Restriction fragment length polymorphism
5.8 Polymorphic sites
5.9 Analysis
6. Conclusions
Objectives & Topics
This research aims to estimate the level of genetic diversity and assess genetic relationships among six common jackfruit varieties in Kerala, India. The study utilizes molecular marker technology, specifically sequence analysis of the rbcL gene, to overcome the limitations of traditional morphological classification and to provide a more reliable basis for plant conservation and breeding strategies.
- Morphological assessment of diverse jackfruit varieties in Kerala.
- Application of molecular markers (RFLP and PCR) for genomic characterization.
- Sequence analysis of the rbcL gene to determine genetic relationships.
- Evaluation of genetic diversity levels to support sustainable livelihood strategies.
Excerpt from the Book
4.6 Isolation of DNA
The total genomic samples from jackfruit leaves were isolated using a modified CTAB method with the addition of β-mercaptoethanol (Simon et al., 2007; Shyamalamma et al., 2008). Three gram leaves samples were wiped with 70% alcohol and chopped into fine pieces and later homognized along with 10 ml extraction buffer using a pre-chilled pestile and motar. The extraction buffer contains 100 mM Tris-HCl, pH 8.0, 20 mM EDTA, 1.4 M NaCl, 3% (w/v) CTAB, 2% polyvinyl pyrrolidine (PVP) and 1% β-mercaptoethanol. The contents were slowly mixed and incubated in water bath at 65°C for 1 hr with slight shaking. The contents were brought to room temperature after incubation and 5 ml chloroform: isoamyl alcohol mixture (24:1) was added. The tubes were centrifuged at 8000 rpm for 20 min at 4°C till a clear supernatant was obtained. After the final spin, the DNA was precipitated using ice-cold isopropanol for overnight at 4°C. The tubes were further centrifuged at 5000 rpm and the pellet washed with 2-3 drops of 70% alcohol, air dried and redissolved in 50 µl Tris-EDTA (TE) buffer and stored at -20°C till further analysis (Simon et al., 2007; Shyamalamma et al., 2008; Xiaoming and Xiuxin, 2001).
Summary of Chapters
1. Introduction: Provides an overview of the jackfruit tree, its taxonomy, and the importance of genetic diversity studies for this underutilized crop.
2. Review of literature: Examines existing studies on jackfruit morphology, traditional uses, and the application of molecular markers in plant breeding.
3. Hypothesis: Outlines the research assumptions regarding genotypic differences and the feasibility of using the rbcL gene for phylogenetic analysis.
4. Materials and Methods: Describes the study area, sampling collection process, morphological descriptors, and detailed laboratory procedures for DNA isolation, PCR, and data sequencing.
5. Results and discussion: Presents the analysis of collected samples categorized into general, leaf, and fruit characteristics, along with molecular findings including sequence alignment and phylogenetic tree construction.
6. Conclusions: Summarizes the key findings, confirming significant morphological variations and validating the use of the rbcL gene as a promising tool for establishing genetic diversity in jackfruit.
Keywords
Anti-oxidant, Jackfruit, rbcL, Monecious, RFLP, Underutilized fruit, Genetic diversity, DNA extraction, PCR, Phylogenetic analysis, Morphology, Kerala, Plant breeding, Molecular markers, Taxonomy
Frequently Asked Questions
What is the primary focus of this publication?
The publication focuses on evaluating the genetic diversity of six jackfruit varieties in Kerala using molecular techniques, specifically rbcL gene sequencing.
Which specific jackfruit varieties were analyzed in this study?
The study analyzed six common varieties: Koozha (Ku), Varikka (Va), Unda chakka (Uc), Pathi varikka (Pv), Theen varikka (Tv), and Singapore varikka (Sv).
What is the main objective of the research?
The main objective is to estimate genetic diversity and assess genetic relationships among these varieties to facilitate better conservation and agricultural management.
Which scientific methods were employed?
The researchers used a combination of field surveys for morphological assessment and advanced genetic methods, including modified CTAB DNA isolation, PCR amplification, and RFLP markers.
What does the main body of the work cover?
The main body covers botanical descriptions, sample collection methods in Kerala, morphological classification parameters, and detailed computational analysis of sequence data.
Which keywords best characterize this work?
Key terms include Genetic diversity, rbcL, Jackfruit, Molecular markers, PCR, RFLP, and Plant breeding.
Why is the rbcL gene used in this phylogenetic analysis?
The rbcL gene is chosen for its slow synonymous nucleotide substitution rate and functional constraints, making it an appropriate marker for inferring phylogenetic relationships at taxonomic levels.
What are the findings regarding the morphological variations?
The study found significant morphological variations in leaf, fruit, and trunk characteristics among the six varieties, suggesting that these differences are genetically determined rather than purely environmental.
- Quote paper
- Dr. Prem Jose Vazhacharickal (Author), Jiby John Mathew (Author), Ajesh C Kuriakose (Author), Sajeshkumar N.K (Author), 2016, Evaluation of genetic diversity of jackfruit (Artocapus heterophyllus Lam) varieties based on sequence analysis of the rbcL gene, Munich, GRIN Verlag, https://www.grin.com/document/351290
