Assessment of genetic diversity through RAPD, ISSR and AFLP markers in Podophyllum hexandrum: a medicinal herb from the Northwestern Himalayan region

Abstract

Total synthesis of podophyllotoxin is an expensive process and availability of the compound from the natural resources is an important issue for pharmaceutical companies that manufacture anticancer drugs. In order to facilitate reasoned scientific decisions on its management and conservation for selective breeding programme, genetic analysis of 28 populations was done with 19 random primers, 11 ISSR primers and 13 AFLP primer pairs. A total of 92.37 %, 83.82 % and 84.40 % genetic polymorphism among the populations of Podophyllum were detected using RAPD, ISSR and AFLP makers, respectively. Similarly the mean coefficient of gene differentiation (Gst) were 0.69, 0.63 and 0.51, indicating that 33.77 %, 29.44 % and 26 % of the genetic diversity resided within the population. Analysis of molecular variance (AMOVA) indicated that 53 %, 62 % and 64 % of the genetic diversity among the studied populations was attributed to geographical location while 47 %, 38 % and 36 % was attributed to differences in their habitats using RAPD, ISSR and AFLP markers. An overall value of mean estimated number of gene flow (Nm) were 0.110, 0.147 and 0.24 from RAPD, ISSR and AFLP markers indicating that there was limited gene flow among the sampled populations.

Genetic diversity and structure of the threatened species Sinopodophyllum hexandrum (Royle) Ying

“…Furthermore, the Mantel test revealed that the genetic distance between populations did not correlate with the geographical distance. Restricted gene flow associated with geographical distance is consistent with the results of previous studies on this species (Alam et al, 2008;Alam et al, 2009;Naik et al, 2010).…”

“…Comparing the average genetic diversity with that of S. hexandrum from other regions (Xiao et al, 2006b;Alam et al, 2008;Naik et al, 2010;Liu et al, 2014) and with other Berberidaceae species (Qiu et al, 2006;Zong et al, 2008) based on the ISSR approach showed that S. hexandrum populations have low genetic diversity (average H E = 0.0366). The S. hexandrum populations from the Northwestern Himalayan region (Xiao et al, 2006b;Alam et al, 2008;Naik et al, 2010) (average H E = 0.2944, 0.092, respectively) and Qinling Mountains (average H E = 0.0621) showed high genetic diversity. S. hexandrum is native to the Himalayan region, where it grows in valleys with secondary vegetation, under shrubs, or around trees (Ma and Hu, 1996).…”

“…9) and the study of S. hexandrum (Xiao et al, 2006a,b;Alam et al, 2008;Alam et al, 2009;Naik et al, 2010;Liu et al, 2014). The reaction system was optimized by screening DNA, Mg 2+ , dNTP, primer (UBC900 was used for preliminary test), Taq DNA polymerase concentrations, annealing temperatures, and reaction conditions.…”

“…However, previous studies have focused on determining the chemical components (Giri and Lakshmi, 2000;Zhao et al, 2011), biological properties (Chattopadhyay et al, 2004), micropropagation (Nadeem et al, 2000), and phylogenetic evolution (Li et al, 2011) of this species. To date, limited publications have characterized the genetic diversity of S. hexandrum, and only studies in populations from the Northwestern Himalayan region are available (Alam et al, 2008;Alam et al, 2009;Naik et al, 2010). Of note, the genetic diversity of this species was investigated in populations from western Sichuan Province and Qinling Mountains in China, using molecular marker techniques (Xiao et al, 2006a(Xiao et al, , 2006bLiu et al, 2014).…”

Genetic Diversity of the Endemic Species Phlomis aurea Decne. in Southern Sinai, Egypt

“…It is also used in the investigation of genetic diversity within and among populations thus provides valid evidence for genetic relatedness (Crawford, The DNA molecular markers are not influenced by the external environmental factors unlike that of morphological and biochemical markers, which had its own limitations as they were always, do not completely represent the genetic structure (Noli et al, 1997) and hence accurately testify the genetic relationship between and among individual groups (Lawrence et al, 1992). Inter-simple sequence repeats (ISSR)has been used regularly for genetic diversity assessment as a thorough knowledge of the level and distribution of genetic variation (of length of DNA fragments between simple sequence repeats), which was essential for conservation (Dreisigacker et al, 2005;Sharma et al, 2008;Naik et al, 2010 andDas et al, 2011). This PCR-based DNA finger printing technique was proven to be very informative and costeffective technique in many individual species as their primers do not require prior knowledge of a species genome (Williams et al, 1990;Zietkiewicz et al, 1994;Kharazian et al, 2015 andBadr et al, 2014;.…”