DNA barcoding of Spiraea hypericifolia L. and development of an efficient micropropagation using nodal explant: an economically important ornamental and medicinal shrub of Eurasian areal
摘要
Spiraea hypericifolia L. (Rosaceae) is an important plant having medicinal importance, and useful for growing on the forest-steppe due to soil strengthening property. However, its natural habitat has been affected due to human activities, and thus development of conservation strategies is needed, but for that correct identification of the species is a prerequisite. Thus, DNA barcoding using matK and rbcL was carried out for S. hypericifolia. The sequenced product size of matK and rbcL was 895 bp and 599 bp, respectively, and they were deposited to GenBank. For shoot regeneration, nodal explants were placed in Murashige and Skoog (MS) medium fortified with 0.5 mg L−1 6-benzyladenine (BA) for bud break. Shoot multiplication was carried out on MS and Linsmaier and Skoog (LS) media containing 0.2 or 0.5 mg L−1 BA or thidiazuron (TDZ), with or without 0.01 mg L−1 indole-3-butyric acid (IBA). Optimal shoot number of 15.00 ± 2.71 with 0.78 ± 0.09 cm shoot length and 6.62 ± 0.65 number of leaves were recorded in LS medium containing 0.5 mg L−1 BA. Rooting of shoots was accomplished on hormone-free ½MS medium, forming 2.57 ± 0.22 number of roots (90% response) with 3.19 ± 0.23 cm root length. The plantlets were transferred to soil:vermiculite (3:1 w/w), where shoot length reached 7.41 ± 0.85 cm with 50.46 ± 7.00 number of leaves, 2.46 ± 0.37 number of roots, and 5.01 ± 0.39 cm root length with 70% survival rate. Plants were successfully hardened and acclimatized in field with 47% survival rate within 2.5 mo. They continued to adapt and grow in field conditions with increased shoot number (6.20 ± 0.68), shoot length (22.17 ± 7.00 cm), and number of leaves per shoot (33.83 ± 11.50) within 10 mo.