Fenugreek is one of the most important spice crops largely grown in Rajasthan which is also termed as “Bowl of Fenugreek” and produces more than 77.5% of the country’s fenugreek. Looking its importance in the Indian economy, the present study was carried out using 20 fenugreek genotypes to characterize them through SSR molecular markers to record precise assessment of genetic diversity for further improvement.
Molecular markers offer plant geneticists and breeders a set of genetic tools that are abundant, non-deleterious, reliable since these are least affected by environmental fluctuation and heritable. In the present investigation, SSR markers were used for the analysis of genetic diversity. DNA was isolated from the young leaf through the CTAB method and analyzes the concentration which ranged between 0.40 µg/µl (RMt-305 and RMt-351) to 2.65 µg/µl (NS 2206-6 and UM 354). The quality of DNA was determined by calculating the ratio between O.D. at 260 nm and O.D. at 280 nm which ranged from 1.78 (Um-353) to 2.05 (RMt-305) indicating fairly high quality of isolated DNA.
In this study 50, RM series SSR primers were screened and only 43 primers produced amplification. Twenty-five primers showed 100% polymorphism with an average of 84.60% polymorphism among the studied genotypes. Forty-three primers amplified a total of 130 alleles, out of which 110 were polymorphic. The average number of alleles per primer was 3.02, while the average numbers of polymorphic alleles per primer were 2.5 (Table 2). A high level of polymorphism was also observed by
Selvaraj et al., (2011) in rice,
Kumari et al., 2013 in
Amaranthus spp.,
Mandal et al., (2018) in lentil,
Suman et al., (2018) in mung bean.
Genetic relationship among fenugreek genotypes and cluster analysis
The similarity coefficient ranged from 0.39 to 0.82 with a maximum similarity coefficient (0.82) was observed between V8 (UM 134) and V9 (UM 152), followed by V1 (NS 2006-1) and V2 (NS 2006-2) which was 0.80. The minimum similarity coefficient (0.39) was observed between V16 (RMt-303) and V17 (RMt-305).
Cluster I which was the major cluster included three genotypes
viz., V1 (NS 2006-1), V2 (NS 2006-2) and V3 (NS 2006-3) at a similarity coefficient of 0.749 (Fig 1). This cluster was divided into two sub-clusters namely A and B. Sub-cluster A had two genotypes
viz., V1 (NS 2006-1) and V2 (NS 2006-2). Cluster II contained five genotypes
viz., V5 (NS 2006-5), V6 (NS 2006-6), V7 (NS 2006-7), V8 (UM 134) and V9 (UM 152) at a similarity coefficient of 0.719. This cluster was divided into two sub-clusters
viz., C and D. Sub-cluster C included only V5 (NS 2006-5) at a similarity coefficient of 0.719, which sub-cluster D consisted of four genotypes
viz., V6 (NS 2006-6), V7 (NS 2006-7), V8 (UM 134) and V9 (UM 152) at a similarity coefficient of 0.762. Cluster III included four genotypes
viz., V11 (UM 189), V13 (UM 353), V12 (UM 202) and V15 (JFG 244) at a similarity coefficient of 0.642. This cluster was divided into two sub-clusters E and F. Sub-cluster E included one genotype
viz.,V11 (UM 189) at a similarity coefficient of 0.642 whereas sub-cluster F consisted of three genotypes
viz., V13 (UM 353), V12 (UM 202) and V15 (JFG 244) at a similarity coefficient of 0.695. Cluster IV included five genotypes
viz., V14 (UM 354), V17 (RMt-303), V18 (RMt-351), V19 (Rmt-1), and V20 (RMt-143) at a similarity coefficient of 0.652. This cluster was divided into two sub-clusters G and H. Sub-cluster G was further sub divided into two sub groups G1 and G2. G1 included single
viz., V14 (UM 354) at a similarity coefficient of 0.715 whereas G2 consisted of two genotypes
viz., V17 (RMt-303) and V18 (RMt-351) at a similarity coefficient of 0.730. Cluster V included three fenugreek entries
viz., V4 (NS 2006-4), V10 (UM 163) and V16 (RMt-303) at a similarity coefficient of 0.508. This cluster was divided into two sub-clusters I and J. Sub-cluster I included one genotype
viz., V4 (NS 2006-4) at a similarity coefficient of 0.508 whereas the sub-cluster II consisted V10 (UM 163) and V16 (RMt-303) at a similarity coefficient of 0.571 (Fig 1).