In-vitro Regeneration of An Endangered Medicinal Plant Anodendron paniculatum (Apocynaceae)

Abstract

Objectives: The present study aims to establish a standardized and efficient regeneration protocol for Anodendron paniculatum A. DC. (Apocynaceae) by optimizing the effect of various plant growth regulators as it is categorized globally as an endangered medicinal plant. Methods: MS medium fortified with 3% sucrose (w/v) and different concentrations of cytokinins 6-benzylamino purine (BAP), kinetin, thidiazuron either alone or in combination with auxins, indole 3-acetic acid (IAA), naphthalene acetic acid (NAA) and GA3. For the in vitro root induction, individual and healthy shoots (3-4 cm in length) were excised in the 7th week and were cultured onto basal and half-strength MS medium supplemented with various concentrations of auxins and incubated in the culture room. Findings: MS medium supplemented with thidiazuron (2.5 𝜇M) in combination with indole 3-acetic acid (2.5 𝜇M) and GA3 (1.25 𝜇M) was most effective, providing multiple shoot regeneration for 79.33% of cotyledonary leaf explants associated with a high number of shoots per explant (20.00 ± 1.15) and length of shoot (2.66 ± 0.08 cm). Whereas, the frequency of response, mean number of shoots, and respective lengths were varied with different concentrations of 6-benzylamino purine when used in combination with IAA or NAA and BAP (5.0 𝜇M) along with IAA (2.5 𝜇M) at the frequency of 47.66%. Whereas, the addition of GA3 (1.25 𝜇M) in combination with BAP (5.0 𝜇M) and IAA (2.5 𝜇M) has shown a maximum number of multiple shoot formation/explant (12.33) with length of shoot (1.53 ± 0.08 cm). MS medium supplemented with 10 𝜇M 6-benzyl amino purine and 2.5 𝜇M naphthalene acetic acid recorded the highest response (53.66%) with more number of the shoot (9.66) and length of the shoot (1.83 ± 0.03 cm) per explant. The maximum percentage of response (84.33%) was recorded at 1 2 strength MS medium with 5.0 𝜇M NAA, whereas the highest number of roots per shoot (6.66 ± 0.33) was induced at MS + 5.0 𝜇M NAA. The highest length of roots (3.66 cm) was recorded at basal MS medium fortified with 5.0 𝜇M NAA and 7.5 𝜇M NAA. Novelty: The standardized protocol method is of first of its kind, a quick and suitable approach for the high quantity production of Anodendron paniculatum with appropriate concentrations of plant growth regulators that meet thecommercial demand of endangered medicinal plants. Keywords: Anodendron paniculatum; Endangered plant; Plant growth regulators; Regeneration; Protocol standardization