Enhanced polyethylene glycol (PEG)–mediated protoplast transformation system for the phytopathogenic fungus, Ganoderma boninense

2021 ◽  
Author(s):  
Fook-Hwa Lim ◽  
Omar Abd Rasid ◽  
Abu Seman Idris ◽  
Abdul Wahab Mohd As’wad ◽  
Ganesan Vadamalai ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Phytopathogenic Fungus ◽  
Ganoderma Boninense ◽  
Protoplast Transformation ◽  
Transformation System

Effect of Heat Shock Treatment on Hordeum Vulgare Protoplast Transformation Mediated by Polyethylene Glycol

2001 ◽  
Vol 44 (1) ◽  
pp. 25-31
Author(s):  
V.K. Tiwari ◽  
J. Zhang ◽  
T.J. Golds ◽  
E.C. Cocking ◽  
M.R. Davey ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Heat Shock ◽  
Hordeum Vulgare ◽  
Shock Treatment ◽  
Heat Shock Treatment ◽  
Protoplast Transformation

Establishment of polyethylene-glycol-mediated protoplast transformation for Lecanicillium lecanii and development of virulence-enhanced strains against Aphis gossypii

2016 ◽  
Vol 72 (10) ◽  
pp. 1951-1958 ◽  
Author(s):  
Yan-Jun Zhang ◽  
Ming Xie ◽  
Xiao-Lin Zhang ◽  
De-Liang Peng ◽  
Wen-Bin Yu ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Aphis Gossypii ◽  
Protoplast Transformation ◽  
Lecanicillium Lecanii

scholarly journals Establishment of a PEG-mediated protoplast transformation system based on DNA and CRISPR/Cas9 ribonucleoprotein complexes for banana

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Shaoping Wu ◽  
Haocheng Zhu ◽  
Jinxing Liu ◽  
Qiaosong Yang ◽  
Xiuhong Shao ◽  
...  
Keyword(s):  
Transient Expression ◽  
Targeted Delivery ◽  
Amplicon Sequencing ◽  
Site Directed Mutagenesis ◽  
Restriction Enzyme Digestion ◽  
Protoplast Transformation ◽  
Transformation System ◽  
Ribonucleoprotein Complexes ◽  
Free Gene ◽  
A Site

Abstract Background To date, CRISPR/Cas9 RNP editing tools have not been applied to the genetic modification of banana. Here, the establishment of a PEG-mediated banana protoplast transformation system makes it possible to build an efficient DNA-free method for a site-directed mutagenesis system. Results Protoplasts constitute a versatile platform for transient expression in plant science. In this study, we established a PEG-mediated banana protoplast transformation system. This system was further optimized for successfully delivering CRISPR/Cas9 and CRISPR/Cas12a plasmids and CRISPR/Cas9 ribonucleoproteins (RNPs) for targeted delivery of the PDS gene into banana protoplasts. Specific bands were observed in PCR-Restriction Enzyme Digestion (PCR-RE) assays, and Sanger sequencing of single clones further confirmed the occurrence of indels at target sites. Deep amplicon sequencing results showed that the editing efficiency of the CRISPR/Cas9 system was higher than that of the other two systems. Conclusions The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana. The application of the CRISPR/Cas9 RNP system enables the generation of banana plants engineered by DNA-free gene editing.

scholarly journals An efficient polyethylene glycol-mediated transformation system of lentiviral vector in Shiraia bambusicola

2016 ◽  
Vol 51 (10) ◽  
pp. 1357-1362 ◽  
Author(s):  
Huaxiang Deng ◽  
Ruijie Gao ◽  
Jiajun Chen ◽  
Xiangru Liao ◽  
Yujie Cai
Keyword(s):  
Polyethylene Glycol ◽  
Lentiviral Vector ◽  
Transformation System ◽  
Shiraia Bambusicola

scholarly journals Draft Genome Sequence of the Phytopathogenic Fungus Ganoderma boninense , the Causal Agent of Basal Stem Rot Disease on Oil Palm

2018 ◽  
Vol 6 (17) ◽  
Author(s):  
Condro Utomo ◽  
Zulfikar Achmad Tanjung ◽  
Redi Aditama ◽  
Rika Fithri Nurani Buana ◽  
Antonius Dony Madu Pratomo ◽  
...  
Keyword(s):  
Fungal Pathogen ◽  
Elaeis Guineensis ◽  
Draft Genome ◽  
Nuclear Genome ◽  
Stem Rot ◽  
Phytopathogenic Fungus ◽  
Ganoderma Boninense ◽  
Basal Stem Rot ◽  
Content Type ◽  
Rot Disease

ABSTRACT Ganoderma boninense is the dominant fungal pathogen of basal stem rot (BSR) disease on Elaeis guineensis . We sequenced the nuclear genome of mycelia using both Illumina and Pacific Biosciences platforms for assembly of scaffolds. The draft genome comprised 79.24 Mb, 495 scaffolds, and 26,226 predicted coding sequences.

scholarly journals Establishment of A PEG-mediated Protoplast Transformation System Based on DNA and CRISPR/Cas9 Ribonucleoprotein Complexes For Banana

2020 ◽  
Author(s):  
Shaoping Wu ◽  
Haocheng Zhu ◽  
Jinxing Liu ◽  
Qiaosong Yang ◽  
Xiuhong Shao ◽  
...  
Keyword(s):  
Transient Expression ◽  
Amplicon Sequencing ◽  
Enzyme Digestion ◽  
Site Directed Mutagenesis ◽  
Plant Science ◽  
Restriction Enzyme Digestion ◽  
Protoplast Transformation ◽  
Transformation System ◽  
Ribonucleoprotein Complexes ◽  
Target Sites

Abstract Background: To date, CRISPR/Cas9 RNPs editing tools have not been applied to genetic modification of banana. Here, the establishment of PEG-mediated banana protoplast transformation system makes it possible to build an efficient DNA- free method for the site-directed mutagenesis system.Results: Protoplasts are a versatile platform for transient expression in plant science. In this study, we established a PEG-mediated banana protoplast transformation system. This system was further optimized for successfully delivering plasmids of CRISPR/Cas9 and CRISPR/Cas12a , CRISPR/Cas9 ribonucleoproteins (RNPs) that targeted PDS gene into banana protoplasts. Specific bands were observed in PCR-Restriction Enzyme Digestion (PCR-RE) assays and monoclonal sequencing further confirmed the occurring of indels at target sites. Deep amplicon sequencing results showed that the editing efficiency of CRISPR/Cas9 system was higher than that of the other two systems. Conclusions: The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana.The application of CRISPR/Cas9 RNPs system enables the generation of DNA-free genome edited banana plants.

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