Isolated microspore culture for embryoid production in Artemisia annua L.

Abstract

The haploidy technique is a useful tool to produce pure, fully homozygous lines in a short time. Artemisia annua (L) is a medicinal plant that produces artemisinin widely known as an anti-malarial drug. Due to its very small ower (< 3.0 mm), this study aimed to conduct microscopic observations to determine the type of owers and their corresponding microspore development stages suitable for microspore embryogenesis and to obtain embryoids from isolated microspore cultures. Media to induce embryoid production were based on NLN medium with 13% or 17% sucrose and the addition of plant growth regulators as follows: (1) a combination of NAA and BA, designated as MCA media; and (2) a combination of 2,4-D and Kinetin, designated as MCAD media. The result indicated that due to their proportion of uninucleate and binucleate microspore stages, owers at about to bloom (AB) and early bloom (EB) stages contained su cient microspore stages of late uninucleate to early binucleate microspores suitable to induce embryogenesis. The production of embryoid is faster in MCA13 and MCA17 than those of MCAD13 and MCAD17. MCA13 and MCAD13 gave rise to abundant callus-like structures than those of MCA17 and MCAD17. It is indicated that the addition of 2.4-D in MCAD media inhibited the growth of the embryoid, due to the formation of a white-gluey structure covered and mixed with the embryoid. Key messageThis is the rst report of successful induction of embryoids in Artemisia annua from isolated microspore culture, which is closely related to ower type and stage of microspore development.