In vitro propagation and phytochemical assessment of Berberis chitria : An important medicinal shrub of Kumaun Himalaya, India

Abstract

Berberis chitria is an important Himalayan shrub known for its diverse medicinal uses. A reproducible in vitro micropropagation protocol for B. chitria has been developed. Seeds were initially inoculated on water agar medium; contamination-free green seeds were transferred to Woody Plant Medium (WPM) with 78.89% seed germination. Initially, maximum multiple shoots (8.30±1.0) per explant were observed in WPM supplemented with 6-benzyl adenine (BA)+α-Naphthalene acetic acid (NAA) (8.88+1.34 µM). The shoot multiplication rate was increased up to 22.89±2.51 shoots per explant on successive subculture but shoot necrosis and concomitant decrease in shoot height was the big problem. Incorporation of casein hydrolysate (CH; 500 mg/L) and gibberellic acid (1.44 µM) in shoot multiplication medium not only facilitated shoot height, but the number of shoots per explant also increased without any necrosis. Microshoots were exposed to auxins in two different ways. The highest rooting percentage (100%) was observed after 8 weeks on WPM supplemented with indol butyric acid (IBA; 100 µM) for 7 days with 5.92±0.36 roots per microshoot. Well-rooted plants were transferred to thermacol pots containing nonsterile, sieved soil and farmyard manure and hardened successfully. This study also provides evidences that the methanolic extract of in vitro raised plants have strong antioxidant activities and higher phenol content than naturally grown plants. Key words: Berberis chitria, 6-benzyl adenine (BA), gibberellic acid (GA3),micropropagation, antioxidant activity.