It is hereby certified that the work described in this thesis entitled "Genetic resources of Curcuma and Zingiber from Northeast India: diversity, characterization and utilization" by Archana Das for the award of the degree of Doctor of Philosophy is an authentic record of the results obtained from the research work carried out under my supervision in the Department of Biotechnology, Indian Institute of Technology Guwahati, India, and this work has not been submitted elsewhere for the award of a degree in any form. My sincere thanks to all the staff members of the Department of Biotechnology, Anita, Niranjan, Nurulda, Prarthana, Rashmiba, Sharan and Krishnada who helped me in all ways when I just asked them.
LIST OF TABLES
ABSTRACT
General introduction
The identification of the members of this genus has traditionally been achieved using morphological data. The members of the genus Zingiber are reported to possess diarylheptaniods in their rhizome essential oils.
Introduction
The green cover of the region is shrinking due to various socio-geographical reasons, causing greater loss of important flora. In this study, an attempt was made to collect, identify and maintain elite genotypes of important members of Zingiberaceae family from NE India.
Literature review
Although only two species were included in this study from the Zingiberaceae flora of Northern India, they nevertheless represent the most commonly encountered genera of the family. Studies on karyotyping and chromosome number in this genus have revealed fewer reports of the existence of polyploidy compared to other members of the family.
Materials and Methods
Sato (1960) carried out karyotypic studies of 24 species belonging to 13 genera of the family Zingiberaceae and found that the base number of the genus Zingiber was n = 11 and Z. Most of the plants wilted very quickly after being cut or dug up. from the ground
Results and Discussion
The preparation of a good slide depends on proper handling of the root material, cutting, maceration and pinching. Adding the acid to the root tip slowly macerates it and also decolorizes the cytoplasmic material, leaving only the chromosomes stained.
Conclusion
Introduction
In vitro culture technique has been recognized as an effective tool for rapid clonal multiplication. However, establishment of tissue culture system is required prior to further exploration of the biosynthetic capabilities of various in vitro cultures.
Literature Review
A considerable amount (10-20%) of rhizomes of the total yield is required for the next crop year (Shirgurkar et al. 2001). Bharalee and his associates (Bharalee et al. 2005) used rhizome bud explants for in vitro clonal propagation of C .
Material and Methods
Total genomic DNA of mother plants and regenerated plants was extracted from fresh tender leaves using the SDS protocol (Kesari et al. PCR amplification of genomic DNA was performed using 10 arbitrary decamer oligonucleotide primers (Operon Tech, USA).The quality and quantity of extracted DNA was checked both by spectrophotometric method and using 1.0% agarose gel prestained with 0.5 µg/ml EtBr.
Results and Discussion
In the same medium, the maximum average length of the longest shoot was also recorded in the MSR medium (4.5 cm) (Figure 2.2D). It was found that a higher concentration of BA inhibited shoot proliferation regardless of the applied Kn concentrations. The maximum average length of the longest shoot in the MSR medium (5.8 cm) was also recorded in the same culture medium (Table 2.5).
Conclusion
However, higher concentrations of sugar source were found to be ideal for in vitro microrhizome production in Z. Application of in vitro propagation techniques can help in the conservation of biodiversity of locally used medicinal plants. The improved in vitro propagation protocol for the conservation of wild but medicinally healthy species of Zingiber from NE India reported here may be useful for the endemic species of Z.
Introduction
The investigation was carried out on two important genera of the family Zingiberaceae which will surely contribute to the conservation effort to ensure the sustainability of nature's wealth for the future. Curcuma and Zingiber are two main genera of this family with great commercial and medicinal significance and the earliest spices known in the east (Singh et al. 2008). Due to overexploitation of such species for their extreme medicinal properties, the germplasm is becoming extinct.
Literature Review
AFLP is considered more powerful and reliable than RAPD for distinguishing genetic diversity (Powell et al. 1996;. Advantages of ISSR markers are that it is accurate, highly polymorphic and can be automated (Medhi et al. 2009). Das et al. al. 2011), but there was no report on any of the endemic Zingiber species from NE India.
Materials and methods
The study also aimed to discuss the usefulness of the three compared methods for assessing genetic diversity. Genomic DNA was isolated from tender leaves (~5 g fresh weight) of the test plants using the modified SDS method of McCouch et al. For RAPD fingerprinting, PCR amplification of the genomic DNA was performed using 20 random decamer oligonucleotide primers for Curcuma and 10 primers for Z.
Results and discussion
In the present study, three DNA-based marker systems were used for the first time to detect the genetic relationships between the nine Curcuma species and ten Z. The RAPD profile generated by OPAN 01 for the 9 different Curcuma species is shown in figure 3.4A. This illustrated a good degree of confidence in the association obtained for the ten ecotypes under study.
Conclusion
Diversity studies generally use different criteria and statistical approaches to obtain a reliable and true picture of genetic diversity (Jatoi et al. 2008). But the use of three sets of marker systems to study the genetic variations in the members of Zingiberaceae is scarce (Das et al. 2011). RAPD markers were found to be useful in distinguishing closely related taxa (Kumar et al. 2009), which was again established in this study.
Antimicrobial spectrum of different solvent based extraction from rhizomes of species of the two genera Curcuma and Zingiber
Introduction
It is estimated that 10% of all plant species are currently threatened in India (Pandey et al. 2005). People in developing countries use traditional medicine for their primary health care needs (Palombo et al. 2001; Cowan 1999). The potential of higher plants as a source for new drugs is therefore still largely unexploited (Dubey et al. 2004).
Literature Review
The genus Curcuma has been extensively studied for its antimicrobial and other biological properties (Negi et al. Boiled rhizome juice has also been used as a remedy for worm infestation in children (Bhuiyan et al. 2009). -inflammatory and anti-HIV activities (Das et al. 1997; Chien et al. 2008).
Materials and Methods
Therefore, another aim of the study was to characterize the essential oils from the rhizome of the selected gingers against gram-positive and gram-negative bacteria using micro-Raman spectroscopy. The effect of the rhizome oil fractions was also investigated by determining the growth kinetics of bacteria. After incubation, 100 µl of the growth mixtures (bacteria + rhizome oil + media) were inoculated onto solid NB-agar (2%) plates and kept at 37 oC for 24 hours.
Results and Discussion
The vegetative plant body and rhizomes of the studied species are shown in Figure 4.1. The growth kinetics of bacterial strains showed a marked decrease in the optical density of bacterial cultures after treatment with rhizome extracts. Therefore, the root oils of the four studied members of the Zingiberaceae are effective against common pathogenic bacteria at a very low concentration of 10 µg/ml.
Antibacterial effect of Z. moran water extract
- Conclusion
Some of the members are also reported to have antimicrobial properties (Sabulal et al. 2006; Norajit et al. 2007). The results suggested that the concentrations of the essential oils from the rhizome were sufficient enough to kill or inhibit the growth of the pathogen. Therefore, an advanced characterization in evaluating the antibacterial potential of rhizome essential oils using micro-Raman spectroscopy is presented here.
Isolation and characterization of potential bioactive compounds from rhizome essential oil of Zingiber moran
5a.1 Introduction
The present study is about one such attempt to investigate one of the endemic plant species of the family Zingiberaceae collected from NE India. The medicinal family Zingiberaceae is found to grow naturally in this region of India, which has been an essential part of ethno-medicinal practices. Therefore, the aim of this chapter was to isolate and characterize at least three potential natural antimicrobial compounds from the rhizome oil of wild ginger (Z. moran).
5a.2 Literature Review
Furthermore, 9 of the 20 best-selling drugs are derived from knowledge of plant secondary metabolites (Tulp and Bohlin 2002). The design and scope of studies should be consistent with traditional use and in consultation with traditional healers. Most herbal products do not have regulatory drug approval to demonstrate their safety and efficacy.
5a.3 Materials and Method
Result and Discussion
Since the crude essential oil contained many components, TLC could not serve as an effective means of separation, and therefore column chromatography was followed to isolate at least three major components of the oil. The initial separation of the three selected components with CC is as shown in Figure 5a.2. The spectra showed single peaks for each of the three isolated compounds, confirming the purity of the same as shown in Figure 5a.3 (A, B & C) for compound 1, 2 and 3, respectively.
Fraction 1, B-Fraction 2, C-Fraction 3
- Conclusion
Linalool or 3,7-dimethyl octa-1,6-dien-3-ol was identified as the third compound from fraction 3 of Z. However, the characterization of the mechanism and the biological activity of chemical constituents is of particular importance. In this study, only three of the chemical constituents from this 'new' ginger variety were isolated and characterized.
5b.2 Literature Review
In vitro and in vivo experimental studies show that these phytochemicals interfere with several cell signaling pathways and lead to apoptosis and cell cycle arrest (Chathoth et al. 2008). Furthermore, since these agents are obtained from natural sources and have been consumed by humans for centuries, they can be considered as safe chemotherapeutic agents (Chathoth et al. 2008). This cancer has been targeted by researchers to discover new anticancer drugs that can replace current unsafe regimens for such a disease (Liu et al. 2008).
5b.3 Materials and methods
