Abstract
Genetic variations of 15 Brahmi (Bacopa monnieri L.) accessions were evaluated using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers. During RAPD analysis, amplification of genomic DNA of the 15 accessions by 22 primers generated 197 fragments, of which 187 were polymorphic with an average of 8.95 bands per primer. The amplified products varied in size from 2,200 to 250 bp. Twenty-five selected ISSR primers produced 284 bands across 15 accessions, of which 270 were polymorphic with an average of 10.80 bands per primer. The PIC value ranges from 0.363 to 0.908 for RAPD primers, while 0.419 to 0.836 in case of ISSR. The size of amplified bands ranged from 2,800 to 240 bp. Similarity index values ranged from 0.16 to 0.95 (RAPD), 0.18 to 0.98 (ISSR) and 0.179 to 0.945 for pooled ISSR and RAPD markers data. Mantel test revealed the similar distribution pattern of the polymorphism between RAPD and ISSR markers and the correlation co-efficient (r) was 0.71384. The results indicated that both of the marker systems RAPD and ISSR, individually or combined can be effectively used in determination of genetic relationship among B. Monnieri accessions collected from different parts of Central India. It could be concluded that the information of genetic similarities and diversity among Brahmi accessions is necessary for their conservation and breeding programs.
Highlights
Bacopa monnieri (L.) Penn. commonly known as ‘NeeraBrahmi’ of family Scrophulariaceae is a small prostrate herb that grows wild in marshy and damp places near water logs throughout India
The inter simple sequence repeats (ISSR) analysis is a very useful molecular tool for studying population genetics on a wide range of plant species, as well as for identifying species, cultivars, or population of the same species (Zietkiewicz et al 1994; Raina et al 2001; Wang et al 2009). In this first report with two PCR-based DNA (RAPD and ISSR) markers, we describe the feasibility for the identification of phylogenetic relationship among B. monnieri accessions collected from Central India
For random amplified polymorphic DNA (RAPD) marker analysis, the PCR reaction mix and program profile were similar to ISSR markers analysis except the annealing temperature which was adjusted to 37 °C
Summary
Bacopa monnieri (L.) Penn. commonly known as ‘NeeraBrahmi’ of family Scrophulariaceae is a small prostrate herb that grows wild in marshy and damp places near water logs throughout India. Medicinal and aromatic plant utilizations and conservation have attracted global attention due to their over exploitation (Chomachalow 1980; Parrotta 2001). According to a report of National Medicinal Plants Board (NMPB) and Technology Information Forecasting and Assessment Council (TIFAC) Department of Science and Technology, Government of India, (2007), the annual market demand for brahmi during 2000 was around 1,000 tones which increased many fold due to its potential uses in ayurvedic system of medicine to treat variety of ailments. B. monnieri has been identified among the seven important medicinal plants recommended for immediate attention and included in the list of highly endangered medicinal plants of India by NMPB and Technology Information Forecasting and Assessment Council (TIFAC), Department of Science and Technology, Government of India (http:// www.nmpb.nic.in/prioritisemedicinalplants.htm). The present status of the plant attracted attention of biotechnologists to conserve this important medicinal herb. Accurate identification based merely on morphological traits becomes difficult and reinforce the importance of molecular markers for precise identification
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