Highly Efficient Genetic Transformation Methods for the Marine Oleaginous Diatom Fistulifera solaris

Naser, Insaf; Yabu, Yusuke; Maeda, Yoshiaki; Tanaka, Tsuyoshi

doi:10.1007/s10126-022-10189-x

Cited by 4 publications

(1 citation statement)

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“…Pre-culture for 3–8 days could improve the GUS expression in microspores [ 36 ]. The results of optimization experiments of Fistulifera solaris showed that the highest conversion rate was achieved by pre-culturing for 48 h [ 37 ]. The above results indicated that the optimal pre-cultivation time varied with individual differences.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Efficient Optimization Method for a Genetic Transformation System of Medicinal Plants Erigeron breviscapus

Zhao

Guo

et al. 2023

IJMS

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Erigeron breviscapus is an important medicinal plant with high medicinal and economic value. It is currently the best natural biological drug for the treatment of obliterative cerebrovascular disease and the sequela of cerebral hemorrhage. Therefore, to solve the contradiction between supply and demand, the study of genetic transformation of E. breviscapus is essential for targeted breeding. However, establishing an efficient genetic transformation system is a lengthy process. In this study, we established a rapid and efficient optimized protocol for genetic transformation of E. breviscapus using the hybrid orthogonal method. The effect of different concentrations of selection pressure (Hygromycin B) on callus induction and the optimal pre-culture time of 7 days were demonstrated. The optimal transformation conditions were as follows: precipitant agents MgCl2 + PEG, target tissue distance 9 cm, helium pressure 650 psi, bombardment once, plasmid DNA concentration 1.0 μg·μL−1, and chamber vacuum pressure 27 mmHg. Integration of the desired genes was verified by amplifying 1.02 kb of htp gene from the T0 transgenic line. Genetic transformation of E. breviscapus was carried out by particle bombardment under the optimized conditions, and a stable transformation efficiency of 36.7% was achieved. This method will also contribute to improving the genetic transformation rate of other medicinal plants.

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Section: Discussionmentioning

confidence: 99%