Development of an In Vitro Propagation Protocol and a Sequence Characterized Amplified Region (SCAR) Marker of Viola serpens Wall. ex Ging

Abstract

An efficient protocol of plant regeneration through indirect organogenesis in Viola serpens was developed in the present study. Culture of leaf explants on MS (Murashige and Skoog) medium supplemented with 2.0 mg/L 6-benzyladenine and 0.13 mg/L 2,4-dichloro phenoxy acetic acid. Adventitious shoot formation was observed when calli were transferred on to MS medium containing 0.5 mg/L α-naphthalene acetic acid and 2.25 mg/L kinetin, which showed the maximum 86% shoot regeneration frequency. The highest root frequency (80.92%) with the 5.6 roots per explant and 1.87 cm root length was observed on MS medium supplemented with 2 mg/L indole-3-butyric acid. The plantlets were transferred to the mixture of sand, coffee husk and soil in the ratio of 1:2:1 in a pot, and placed under 80% shade net for one month. It was then transferred to 30% shade net for another one month, prior to transplantation in the field. These plantlets successfully acclimatized under field conditions. A Sequence Characterized Amplified Region (SCAR) marker was also developed using a 1135 bp amplicon that was obtained from RAPD (Random Amplification of Polymorphic DNA) analysis of six accessions of V. serpens. Testing of several market samples of V. serpens using the SCAR marker revealed successful identification of the genuine samples of V. serpens. This study, therefore, provides a proficient in vitro propagation protocol of V. serpens using leaf explants and a SCAR marker for the authentic identification of V. serpens. This study will be helpful for conservation of authentic V. serpens.

<i>In Vitro</i> Propagation and Cytological Analysis of <i>Boerhaavia diffusa </i>L—An Important Medicinal Plant Species of Bangladesh

“…The highest elongation (4.24 ± 0.06 cm) was observed in the medium fortified with 3.0 mg/l BAP + 0.5 mg/l NAA (Figure 1(c) & Figure 1(d)) after 30 days of inoculation. The effect of BAP in combination with NAA has been noted in many other medicinal plant species viz Plectranthus amboinicus [13], Orthosiphon stamineus [14] and Viola serpens [15].…”

An overview of remote monitoring methods in biodiversity conservation

“…SCAR markers provide authenticate information both for species identification and population genetic diversity analysis. Researchers have successfully developed SCAR markers for the medicinal plant V. serpens using 1135-bp long amplicon through RAPD obtained by six accessions of the plant, thereby preventing it from extinction (Jha et al 2020 ) (Table 3 , Fig. 2D ).…”

Development and Application of a Cultivar-Specific Sequence-Characterized Amplified Region (SCAR) Marker for the Detection of Chrysanthemum morifolium Ramat. ‘Daboju’

“…To date, many DNA molecular markers, including randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), inter-simple sequence repeat (ISSR), sequence-related amplified polymorphism (SRAP), and simple sequence repeat (SSR), have been used widely used in genetic diversity assessment [ 20 , 21 , 22 ], phylogenetic analysis [ 23 ], genetic linkage map construction [ 24 ], QTL mapping [ 25 , 26 ], and phenotypic trait association analysis [ 27 ] of chrysanthemum germplasm resources. However, a number of studies have shown that the direct application of these markers to the molecular identification of plants is not ideal [ 28 , 29 , 30 ]. Sequence-characterized amplified region (SCAR), a class of reliable PCR-based DNA molecular marker, has been developed from a specific nucleotide sequence generated by certain traditional molecular markers, such as RAPDs, ISSRs, and AFLPs [ 31 , 32 , 33 ].…”

“…Sequence-characterized amplified region (SCAR), a class of reliable PCR-based DNA molecular marker, has been developed from a specific nucleotide sequence generated by certain traditional molecular markers, such as RAPDs, ISSRs, and AFLPs [ 31 , 32 , 33 ]. SCAR markers have been widely used in the molecular identification of plant species, cultivars, or traditional medicinal preparations due to their high stability and specificity, simple operation, and low cost [ 28 , 30 , 34 , 35 , 36 , 37 ].…”