In this study, the callus initiation and rhizogenesis of Justicia gendarussa from nodal and leaf explants were established in Murashige and Skoog medium supplemented with different hormonal concentrations and also optimization of callus initiation at hormonal concentrations. As a result the callus was initiated on MS medium supplemented with different hormones. The higher concentration of NAA and BA induced callus from nodal and leaf explants. The higher concentrations of 2, 4 – D and kinetin induced callus on nodal explants. Lower concentration of 2, 4 – D and kinetin induce callus on leaf explants. The combination of NAA and kinetin induce rhizogenesis from nodal explants. The antimicrobial activity of callus was evaluated by well diffusion method. Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilus, Staphylococcus aureus, Klebsiella pneumonia were used as test organisms. The results showed that Justicia gendarussa have antimicrobial activity on Escherichia coli, Klebsiella pneumonia, and Staphylococcus aureus.
Keywords: Justicia gendarussa, callus initiation, direct organogenesis, antimicrobial assay.
Table of Contents
Optimization of media formulations for callus induction and direct organogenesis in Justicia gendarussa Burm Fil., an exotic medicinal plant in Kerala
Abstract
1. Introduction
1.1 Aim
1.2 Objectives
2. Review of literature
2.1 Justicia gendarussa Burn Fil. Common names
2.2 Taxonomical classification: Justicia gendarussa Burn Fil.
3. Hypothesis
4. Materials and Methods
4.1 Study area
4.2 Plant material
4.3 Equipment’s used for experiment
4.4 Culture media
4.5 Compositions of MS media (Chawla H.S.)
4.6 Preparation of MS media (stock preparation)
4.7 Compositions of nutrient broth
4.8 Preparation of nutrient broth
4.9 Compositions of MHA
4.10 Preparation of MHA
4.11 Test organisms
4.12 Experimental systems
4.13 Preparation of MS media
4.14 Preparation of explants
4.15 Surface sterilization of explants
4.16 Inoculation procedure
4.17 Statistical analysis
5. Results and discussion
5.1 Establishment of callus cultures from nodal and leaf explants
5.2 Establishment of direct organogenesis from nodal explants
5.3 Optimization of callus initiation of Justicia gendarussa
5.4 Evaluation of antimicrobial activity
6. Conclusions
References
Research Objectives and Topics
The research aims to develop an efficient and reproducible in vitro protocol for callus proliferation and direct organogenesis of the medicinal plant Justicia gendarussa, while simultaneously evaluating the antimicrobial properties of the generated callus tissue against various human pathogens.
- In vitro plant tissue culture and callus induction techniques.
- Optimization of hormonal concentrations (NAA, BA, 2,4-D, Kinetin) for growth.
- Direct organogenesis from nodal explants of Justicia gendarussa.
- Antimicrobial activity evaluation using the agar well diffusion method.
- Phytochemical considerations for medicinal applications.
Excerpt from the book
4.16.4 Inoculation
Under aseptic conditions, explants were inoculated on basal MS (Murashige and Skoog) (1962) medium containing 3% (w/v) sucrose, supplemented with four different auxins (2, 4-D, NAA, IAA, IBA) and cytokinine (BA). The pH was adjusted to 5.8 prior to the addition of 0.8% agar and autoclaved at 121°C for 15 minutes. Cultures were then incubated at 20 ± 2°C with a 16-hour photoperiod by cool white fluorescent tubes (Das et al. (2010) and 70-75% relative humidity (Mukherjee et al. (1991).
Summary of Chapters
1. Introduction: Outlines the fundamentals of plant tissue culture and introduces Justicia gendarussa as a significant medicinal plant used in traditional treatments.
2. Review of literature: Provides a comprehensive overview of existing studies on the classification, medicinal properties, and in vitro regeneration of Justicia species.
3. Hypothesis: Proposes that the growth response and organogenesis in Justicia gendarussa are dependent on varying hormonal concentrations.
4. Materials and Methods: Details the experimental setup, including media preparation (MS and MHA), sterilization processes, and the specific procedures for inoculation and statistical analysis.
5. Results and discussion: Presents the findings regarding the success of callus induction, direct organogenesis, and the observed antimicrobial efficacy of the callus extracts.
6. Conclusions: Summarizes the study’s findings and suggests that the optimized hormonal combinations are effective for in vitro propagation and antimicrobial production.
Keywords
Justicia gendarussa, callus initiation, direct organogenesis, antimicrobial assay, Murashige and Skoog medium, plant growth regulators, rhizogenesis, nodal explants, leaf explants, in vitro propagation, phytochemical analysis, Ayurveda.
Frequently Asked Questions
What is the primary focus of this research?
The research focuses on the optimization of culture media for callus induction and direct organogenesis of the medicinal plant Justicia gendarussa, as well as testing its antimicrobial activity.
What are the central themes of the work?
The study revolves around plant biotechnology, specifically plant tissue culture, the impact of various growth hormones, and the evaluation of medicinal plant extracts for antimicrobial potential.
What is the primary objective of this study?
The primary goal is to develop a simple, efficient, and reproducible in vitro protocol for callus proliferation and organogenesis of Justicia gendarussa in Kerala.
Which scientific methods were employed?
The researchers used Murashige and Skoog (MS) media for cultivation, combined with different concentrations of plant hormones (NAA, BA, 2,4-D, Kinetin) and assessed antimicrobial activity via the agar well diffusion method.
What topics are covered in the main body?
The main body details the preparation of culture media, surface sterilization techniques, inoculation procedures, and discusses the observed growth responses and antimicrobial zones of inhibition.
Which keywords define this work?
Key terms include Justicia gendarussa, callus initiation, direct organogenesis, antimicrobial assay, and in vitro propagation.
Which specific plant parts were used for the experiment?
The study utilized young leaves and nodal regions of Justicia gendarussa as explants for the tissue culture experiments.
What were the findings regarding antimicrobial activity?
The callus extracts showed significant antimicrobial activity against Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus.
How was the hormonal concentration optimized?
Optimization was achieved by testing various combinations of auxins and cytokinins to determine which concentrations induced the most prolific callus growth and organogenesis.
What is the significance of the findings in chapter 5?
Chapter 5 demonstrates that specific combinations, such as 0.7mg/L NAA + 0.2 mg/L BA, significantly improve callus initiation compared to other concentrations.
- Citar trabajo
- Dr. Prem Jose Vazhacharickal (Autor), Elizabeth Cherian (Autor), Noby Mathew (Autor), 2017, Optimization of Media Formulations for Callus Induction and direct Organogenesis in Justicia Gendrussa Burm Fil, Múnich, GRIN Verlag, https://www.grin.com/document/378387
