Abstract
Kalmegh (Andrographis paniculata (Burm. F.) Nees) is one of the most important medicinal plants and has been widely explored as traditional medicine. To exploit its natural genetic diversity and initiations of molecular breeding to develop novel cultivars or varieties, developments of genomic resources are essential. Four microsatellite-enriched genomic libraries—(CT)14, (GT)12, (AG)15 and (AAC)8—were constructed using the genomic DNA of A. paniculata. Initially, 183 recombinant colonies were screened for the presence of CT, GT, AG, and AAC microsatellite repeats, out of which 47 clones found positive for the desired simple sequence repeats (SSRs). It was found that few colonies had more than one desirable SSR. Thus, a sum of 67 SSRs were designed and synthesized for their validation among 42 A. paniculata accessions. Out of the 67 SSRs used for genotyping, only 41 were found to be polymorphic. The developed set of g-SSR markers showed substantial genetic variability among the selected A. paniculata accessions, with an average polymorphic information content (PIC) value of 0.32. Neighbor-joining tree analysis, population structure analysis, analysis of molecular variance (AMOVA), and principal coordinate analysis (PCoA) illustrated the considerable genetic diversity among them. The novel g-SSR markers developed in the present study could be important genomic resources for future applications in A. paniculata.
Highlights
The Kalmegh is an important medicinal crop species that is botanically known as Andrographis paniculate
Seeds of 42 A. paniculata accessions were collected from the National Gene Bank Indian Council of Agricultural Research (ICAR)-NBPGR, New Delhi, which were earlier collected from different geographical regions of Indian states (Figure 1)
A set of 67 developed simple sequence repeats (SSRs) primers were selected for genetic diversity analysis, and these primers were amplified on 42 A. paniculata accessions, out of which 41 were found reproducible and polymorphic
Summary
The Kalmegh is an important medicinal crop species that is botanically known as Andrographis paniculate Andrographolide is one of the major bitter-tasting secondary metabolites derived from Kalmegh, a major bioactive substance responsible for the therapeutic interest [6] This crop species is widely used as a traditional medicine in different parts of the world due to its versatile biological properties like immune-stimulatory [7], hepatoprotection [8], antibacterial [9], antimalarial [10], antithrombotic [11], antitumor [12], and anti-inflammatory [13]. These dominant marker systems have low reproducibility and low consistency [18], which is a significant impediment for their further utilization in A. paniculate genomics and molecular breeding These dominant marker systems, namely, microsatellite or simple sequence repeat (SSR) markers, are one of the most preferred marker systems in studies of plant breeding and genetics due to the abundance in genomes, codominant natures, high reproducibility, multiallelic traits, and high transferability across the species [19].
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