Isolated microspore culture for embryoid production in Artemisia annua L.

IF 2.3 3区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Plant Cell, Tissue and Organ Culture Pub Date : 2024-03-20 DOI:10.1007/s11240-024-02716-6

Ragapadmi Purnamaningsih, Iswari Saraswati Dewi, Deden Sukmadjaja, Aniversari Apriana, Bambang Sapta Purwoko

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Abstract

The haploidy technique is a useful tool for quickly producing pure, fully homozygous lines. Artemisia annua L. is a medicinal plant that produces artemisinin, a widely used antimalarial drug. Because of its extremely small flowers (≤ 3.0 mm), this study conducted microscopic observations to determine the types of flowers of A. annua suitable for microspore embryogenesis, as well as their corresponding microspore development stages, and obtained embryoids (non-zygotic embryos) from isolated microspore cultures. The media for inducing embryoid production were based on Nitsch and Nitsch medium containing 13% or 17% sucrose and the following plant growth regulators: (1) a combination of naphthaleneacetic acid and 6-benzyladenine (MCA medium) and (2) a combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (MCAD medium). The results indicated that based on the proportions of uninucleate and binucleate microspores, flowers at the prebloom and early bloom stages contained sufficient late uninucleate to early binucleate microspores suitable for inducing embryogenesis. The production of microspore-derived embryogenic (MDE) structures was faster in MCA13 and MCA17 media than in MCAD13 and MCAD17 media. MCA13 and MCAD13 media induced the production of more callus-like structures than MCA17 and MCAD17 media. Thus, the addition of 2, 4-D to MCAD medium inhibited the growth of MDE structures. Globular embryoids emerged from the multicellular cluster.

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用于黄花蒿胚状体生产的分离小孢子培养

摘要单倍体技术是快速培育纯合、完全同源品系的有效工具。黄花蒿（Artemisia annua L.）是一种生产青蒿素的药用植物，青蒿素是一种广泛使用的抗疟疾药物。由于其花朵极小（≤ 3.0 毫米），本研究通过显微镜观察确定了适合小孢子胚胎发生的黄花蒿花朵类型及其相应的小孢子发育阶段，并从分离的小孢子培养物中获得了胚状体（非同源胚胎）。诱导胚状体产生的培养基是基于含有 13%或 17%蔗糖和以下植物生长调节剂的 Nitsch 和 Nitsch 培养基：(1) 萘乙酸和 6-苄基腺嘌呤的组合（MCA 培养基）；(2) 2,4-二氯苯氧乙酸（2,4-D）和激肽的组合（MCAD 培养基）。结果表明，根据无核小孢子和双核小孢子的比例，开花前和开花初期的花朵含有足够的适合诱导胚胎发生的晚期无核小孢子和早期双核小孢子。与 MCAD13 和 MCAD17 培养基相比，在 MCA13 和 MCA17 培养基中产生小孢子胚胎发生（MDE）结构的速度更快。与 MCA17 和 MCAD17 培养基相比，MCA13 和 MCAD13 培养基能诱导产生更多的胼胝体样结构。因此，在 MCAD 培养基中添加 2, 4-D 会抑制 MDE 结构的生长。多细胞团中出现球状胚。

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来源期刊

Plant Cell, Tissue and Organ Culture 生物-生物工程与应用微生物

CiteScore

5.40

自引率

13.30%

发文量

203

审稿时长

3.3 months

期刊介绍： This journal highlights the myriad breakthrough technologies and discoveries in plant biology and biotechnology. Plant Cell, Tissue and Organ Culture (PCTOC: Journal of Plant Biotechnology) details high-throughput analysis of gene function and expression, gene silencing and overexpression analyses, RNAi, siRNA, and miRNA studies, and much more. It examines the transcriptional and/or translational events involved in gene regulation as well as those molecular controls involved in morphogenesis of plant cells and tissues. The journal also covers practical and applied plant biotechnology, including regeneration, organogenesis and somatic embryogenesis, gene transfer, gene flow, secondary metabolites, metabolic engineering, and impact of transgene(s) dissemination into managed and unmanaged plant systems.