In vitro micropropagation of Basilicum polystachyon (L.) Moench and identification of endogenous auxin through HPLC

Abstract

An efficient plant regeneration protocol was developed for Basilicum polystachyon (L.) Moench using shoot tip from in vitro germinated plant. Both shoot multiplication and root induction were initiated from shoot tip explants in Murashige and Skoog’s (MS) basal medium supplemented with N6-benzylaminopurine (BAP) and 6-furfurylaminopurine (Kin) combination with 1-naphthaleneacetic acid (NAA) and without any plant growth regulator. Among the different concentrations and combinations of growth regulators, the highest number of shoots per explants was induced on 13.32 μM BAP with 0.53 μM NAA. It was also found that the multiplication of shoots along with roots induced in MS medium without any plant growth regulators. The in vitro grown plants were successfully hardened and acclimatized in the field with a 99% survival rate. The results obtained from HPLC analysis established the presence of a significant amount of endogenous auxin, viz. indole-3-acetic acid acid and indole-3-butyric acid in the shoot and root tips of B. polystachyon. This is the first report of a successful multiplication of B. polystachyon in absence of plant growth regulators and the presence of an abundant quantity of endogenous auxin in root and shoot tips using Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) coupled with ultraviolet–visible (UV–Vis) detector. This is the first report on identification of endogenous indole-3-acetic acid and indole-3-butyric acid from the shoot and root tips of B. polystachyon (L.) Moench.