Genetic and chemo-divergence in eighteen core collection of Zingiber officinale from North-West Himalayas☆
Introduction
Zingiberaceae family harbors many commercially important plants including Zingiber officinale (Ginger). Rhizome of the plant is used as spice, flavorant and herbal medicine for anti-emetic, antipyretic and hypotensive effects, since ancient time (Govindarajan, 1982). Due to its universal appeal and adaptability, ginger is cultivated in most tropical and subtropical countries (Ravindran and Nirmal, 2005). According to WHO, India is among the largest cultivator and exporter of ginger and ginger derived product globally, occupying 50% of world market in terms of ginger oil and oleoresin (WHO, 1999). The crop is grown throughout India from temperate hilly regions to subtropical wet areas. Cultivation is mostly based on developed varieties and cultivars or landraces selected by farmers over time, which are well adapted to local needs and prevailing environmental conditions. Most of these germplasm are grown by farmers over small marginal areas generally for self or local consumptions (Govindarajan, 1982, Ravindran and Nirmal, 2005).
Beside uses in medicine, ginger rhizome is the source of essential oil which is valued for its pleasant aroma ranging from sweet, warm to camphoraceous or citrus-like depending on the relative essential oil constituents which decide the quality and industrial usage of the oil (Gupta et al., 2011). Literature survey revealed hydro-distilled volatile oil of ginger is a combination of monoterpenes, oxygenated monoterpenes and sesquiterpenes, whose relative proportion depends on genotype and influenced by geographical locations (Sharma et al., 2002, Raina et al., 2005). In India, the oil obtained from the southern part of country is rich in sesquiterpenes and oxygenated sesquiterpenes whereas that from Northern India has more of oxygenated monoterpenes (Figueiredo et al., 2008, Gupta et al., 2011). Since environmental factors, in addition to genetic variation influence phyto-chemical profile, the chemical and molecular characterization becomes necessary for proper characterization of a medicinal plant.
Polymerase Chain Reaction (PCR) based techniques provide a simple, useful and definite method of molecular characterization, in contrast to chemical and morphological markers which are unreliable and affected by the environmental factors. These markers are reproducible, robust, polymorphic, dominant/co-dominant and unaffected by age thereby lowering the cost, time and labor for analyses (Zietkiewicz et al., 1994, Kalender et al., 1999). Molecular characterization using various molecular markers (ISSR, SSR, IRAP) have been successfully used in estimating the extent of genetic diversity at inter/intra-specific and varietal levels of a number of species in a wide range of crop from angiosperm (Joshi et al., 2000, Charters and Wilkinson, 2000, Charles et al., 2008) to gymnosperms (Yong et al., 2003). They have been also utilized in assessing genetic variation either specifically in Zingiber or more general in Zingiberaceae family (Jatoi et al., 2008, Kizhakkayil and Sasikumar, 2010). However, most of the earlier work published included genotypes from East Asian countries with medium to low genetic diversity.
Despite immense commercial importance of the crop, studies pertaining to essential oil variation and molecular genotyping of this clonally propagated crop are sparse (Jiang et al., 2006, Kavyashree, 2008) and no study pertaining to chemical-genetic relationship have been documented on germplasm utilized/available in North-Western parts of India. In the present study, ginger rhizome was collected from seven locations of North Western Himalayan India, which were being cultivated by the farmers of the two states. They are best adapted to the local climate and grown for local needs of the area. These germplasm were selected on the basis of differences in rhizome color, size and aroma. Genetic study was performed using three molecular marker techniques ISSR, SSR and IRAP and investigation was further extended to rhizome essential oil analysis to determine the variation in essential oil components.
Section snippets
Plant material
Ginger germplasm was collected from farmer's field located in Uttrakhand and Himachal Pradesh, the two states of North-West Himalayan region of India and cultivated and maintained at IIIM Jammu experimental farm during 2009–2010. The initial criteria of selection were based on visual properties like rhizome size, color, odor and fiber level. North-West Himalayan region includes hilly terrain with temperate climate of Himachal Pradesh, (32°29′N:75°10′E) and Uttarakhand (30°15′N:79°15′E), India.
Results
Morphologically, more than half of the plants (11) were of medium height and rhizome size was observed to be large in 13 of the 18 samples studied, while more than 66% of the rhizomes were of light brown color, in the present investigation (Table 1).
Discussion
Although di- and tri-nucleotide motifs primed equal number of polymorphic loci/primer (8), polymorphism exhibited by tri-nucleotide motif was maximum (85.7%) while it was minimum (69.6%) in di-nucleotide repeat sequence motif. Thus choice of primer plays a crucial role in assessing germplasm diversity. Although a higher number of ISSR primers need to be analyzed, our results indicated tri-nucleotide repeat sequences are present more abundantly in ginger genome. Similar to our findings, Galvan
Conclusions
In summary, the results demonstrated unexplored chemical as well as genetic diversity existing in the North Western Himalayan region. It also revealed efficiency of ISSR, SSR and IRAP genetic marker techniques and oil based chemical diversity in distinguishing ginger germplasm at an intra-species level and in revealing the genetic make-up of population collected from North-West Himalayan states of India. Molecular and essential oil analyses, in combination with advanced computational methods
Acknowledgements
Authors are thankful to the Director, Ram Vishwakarma, Indian Institute of Integrative Medicine (CSIR), Jammu for providing the facilities. Acknowledgements are also due to Department of Biotechnology for funding the study (BT/PR9218/AGR/05/376/2007). Mr. Pankaj Pandotra and Dr. Mohd. Kashif Husain extends acknowledgements to CSIR for providing the fellowships.
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MS Communication No. IIIM/1551/2013.