Germline transformation of Artemisia annuaL. plant via in planta transformation technology “Floral dip”

Q1 Immunology and Microbiology

Biotechnology Reports Pub Date : 2022-12-01 DOI:10.1016/j.btre.2022.e00761

Firdaus Qamar, Shazia Khan, Kudsiya Ashrafi, Sadia Iqrar, Syed Naved Quadri, Monica Saifi, M.Z. Abdin

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引用次数: 1

Abstract

The therapeutic efficacy of Artemisia annua L. is governed by artemisinin (ART), prevalently produced by A. annua extraction. Due to the modest amount of ART (0.01-1 %dw) in this plant, commercialization of ACTs is difficult. In this study, the floral-dip based transformation protocol for A. annua was developed to enhance expression of artemisinin biosynthesis genes and ART content. For dipping, the effective infiltration media components were optimized, and to obtain high transformation (26.9%) partially open bud stage capitulum of floral development was used. Hygromycin phospho-transferase (hptII) selection marker was used to validate the transformed T₁ progenies. The copy numbers of the transgene (hptII) in T₁ progenies were determined using a sensitive, high-throughput SYBR Green based quantitative RT-PCR. The results of the hptII transgene were compared with those of the low copy number, internal standard (hmgr). Using optimised PCR conditions, one, two and three transgene copies in T₁ transformants were achieved.

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黄花蒿的种系转化。植物通过植物转化技术“花浸”

黄花蒿(Artemisia annua L.)的治疗效果受青蒿素(artemisinin, ART)的控制，青蒿素通常由黄花蒿提取物产生。由于该厂的ART含量有限(每天0.01- 1%)，ACTs的商业化很困难。为了提高青蒿素生物合成基因的表达和抗逆转录病毒(ART)的含量，本研究开发了基于花浸液的青蒿素转化方案。对于浸渍，优化了有效的浸渍介质成分，利用部分开放的花蕾期头状花序获得较高的转化率(26.9%)。用湿霉素磷酸转移酶(hptII)选择标记对转化后的T1后代进行验证。采用敏感、高通量的SYBR Green定量RT-PCR技术测定T1后代中转基因(hptII)的拷贝数。并与低拷贝数内标(hmgr)的结果进行了比较。利用优化的PCR条件，在T1转化子上分别获得了1、2和3个转基因拷贝。

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来源期刊

Biotechnology Reports Immunology and Microbiology-Applied Microbiology and Biotechnology

CiteScore

15.80

自引率

0.00%

发文量

审稿时长

55 days

期刊介绍： Biotechnology Reports covers all aspects of Biotechnology particularly those reports that are useful and informative and that will be of value to other researchers in related fields. Biotechnology Reports loves ground breaking science, but will also accept good science that can be of use to the biotechnology community. The journal maintains a high quality peer review where submissions are considered on the basis of scientific validity and technical quality. Acceptable paper types are research articles (short or full communications), methods, mini-reviews, and commentaries in the following areas: Healthcare and pharmaceutical biotechnology Agricultural and food biotechnology Environmental biotechnology Molecular biology, cell and tissue engineering and synthetic biology Industrial biotechnology, biofuels and bioenergy Nanobiotechnology Bioinformatics & systems biology New processes and products in biotechnology, bioprocess engineering.