Citrus, one of the major genes of Rutaceae family and most economically important fruit tree and widely cultivated throughout the country. The Citrus have high nutritional value and medicinal value. The three varieties obtained from various districts in Kerala were used in this study. The phylogenetics is the study of the evolutionary relationships among different species which have common ancestor. These relationships are shown in the form of phylogenetic trees composed of branches which indicate the descendents and nodes which represent the most recent common ancestors.
In order to assess the phylogenetic relationships among the 6 samples belongs to the 3 Citrus species (Citrus aurantifolia, Citrus maxima, Citrus limon) a chloroplast gene rbcL and matK was successfully amplified and sequenced. For this study DNA was extracted by using CTAB method. This extracted DNA was analysed by spectrophotometry method for checking purity of DNA.The samples were gel electrophoresed by 1% agarose gel electrophoresis at 80 volts. After electrophoresis the gel is examined in gel documentation system. The DNA band was observed under UV light looking florescent Orangish red colour. The extracted DNA was amplified by PCR method and PCR sample was applied for electrophoresis for checking DNA bands. PCR sample were purified sent for sequencing. Sequences obtained were subjected to editing and alignment using ClustalW programme of Bio Edit and phylogenetic analysis was performed using MEGA software.
High similarity observed between selected verities. The locally collected gene sequence-based phylogeny presented here provides support for the early studies of speciation within the Rutaceae. An understanding of the main phylogenetic relationships between Citrus species will help to fine-tune the taxonomy of Rutaceae.
Table of Contents
- Studies on genetic relationships among locally cultivated Citrus varieties in Kerala employing matk and rbcL gene using PCR technique and RFLP markers.
- Abstract
- Introduction
- Objectives
- Scope of the study
- Taxonomical classification
- Review of literature
- Uses and importance
- Molecular markers
- Hypothesis
- Materials and Methods
- Study area
- Sample collection
- Description of the species
- Citrus aurantifolia/lime:
- Citrus maxima/pomelo:
- Citrus limon/lemon:
- Species identification and morphological characterization
- Isolation of DNA
- Quantification of DNA
- PCR primers
- PCR amplification
- Data sequencing
- Sequence alignment
- Phylogenetic and molecular evolutionary analysis
- Statistical analysis
- Results and discussion
- Blast search for rbcL gene of Citrus species
- Blast search for matk gene of Citrus species
- Sequence alignment
- Construction of phylogentic tree
- Phylogenetic tree using matk gene
- Calculation of the genetic distance
- Conclusions
- Acknowledgements
- References
Objectives and Key Themes
The study aims to investigate the genetic relationships among locally cultivated Citrus varieties in Kerala, India. This is achieved by analyzing the matk and rbcL gene sequences of these varieties using PCR and RFLP techniques. The study focuses on identifying genetic diversity and understanding the evolutionary relationships among different Citrus species. This research contributes to the development of sustainable citrus cultivation practices in Kerala.
- Genetic relationships among locally cultivated Citrus varieties in Kerala
- Analysis of matk and rbcL gene sequences using PCR and RFLP techniques
- Identification of genetic diversity among Citrus species
- Understanding evolutionary relationships among different Citrus species
- Sustainable citrus cultivation practices in Kerala
Chapter Summaries
The study commences with an introduction outlining the objectives, scope, and taxonomical classification of the research. This is followed by a review of literature, exploring the uses and importance of Citrus species, as well as the significance of molecular markers in genetic analysis. The research methodology section details the study area, sample collection process, species description, DNA isolation and quantification, PCR techniques, sequence alignment, and phylogenetic analysis.
The results and discussion section delve into the analysis of BLAST search results for both rbcL and matk genes, sequence alignment, and the construction of phylogenetic trees. Finally, the study concludes with a summary of the findings and their implications.
Keywords
This research focuses on analyzing genetic relationships among locally cultivated Citrus varieties in Kerala using molecular markers such as matk and rbcL genes. The study utilizes PCR and RFLP techniques, focusing on identifying genetic diversity and understanding evolutionary relationships among different Citrus species. This research provides insights into the genetic diversity of Citrus species in Kerala, potentially contributing to improved citrus cultivation practices.
- Citation du texte
- Dr. Prem Jose Vazhacharickal (Auteur), Sajeshkumar N. K. (Auteur), Jiby John Mathew (Auteur), Jaina James (Auteur), 2016, Studies on genetic relationships among locally cultivated Citrus varieties in Kerala employing matK and rbcL gene using PCR technique and RFLP markers, Munich, GRIN Verlag, https://www.grin.com/document/370769
