Author Correction: An in planta biolistic method for stable wheat transformation

Hamada, Hideaki; Linghu, Qianyan; Nagira, Yozo; Miki, Ryuji; Taoka, Naoaki; Imai, Ryozo

doi:10.1038/s41598-017-17188-2

Cited by 5 publications

(3 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It allows the design of viruses capable of producing Cas proteins along with gRNAs contained in viruses with large cargo sizes, enabling their utilization in future research endeavors [ 58 ]. Additionally, recent studies have attempted to improve transformation efficiency by employing morphogenic genes, establishing transformation through phytohormone-free tissue culture [ 19 , 59 ], and performing plant transformation directly, such as in planta transformation, for obtaining shoots directly from plants, without tissue culture [ 16 , 19 ].. The dual virus system may help facilitate efficient gene editing in a wider range of crop species and genotypes, particularly when one viral vector expresses CRISPR components and the other virus expresses morphogenic genes [ 19 , 59 , 60 ].…”

Section: Discussionmentioning

confidence: 99%

“…Efforts have been undertaken to circumvent the requirement of tissue culture by specifically targeting meristem or egg cells for CRISPR/Cas reagents delivery. Recent studies have prioritized the enhancement of transformation efficiency through the utilization of morphogenic genes, the establishment of phytohormone-free tissue culture protocols, and the implementation of direct plant transformation techniques, including in planta transformation [ 16–21 ]. These advanced approaches aim to optimize the effectiveness and efficiency of GE while minimizing dependence on conventional tissue culture methods.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of virus-induced genome editing methods in Solanaceous crops

Lee,

Kang,

Venkatesh

et al. 2023

Horticulture Research

View full text Add to dashboard Cite

Genome editing (GE) using CRISPR/Cas systems has revolutionized plant mutagenesis. However, conventional transgene-mediated GE methods have limitations due to the time-consuming generation of stable transgenic lines expressing the Cas9/single guide RNA (sgRNA) module through tissue cultures. Virus-induced genome editing (VIGE) systems have been successfully employed in model plants, such as Arabidopsis thaliana and Nicotiana spp. In this study, we developed two VIGE methods for Solanaceous plants. First, we used the tobacco rattle virus (TRV) vector to deliver sgRNAs into a transgenic tomato (Solanum lycopersicum) line of cultivar Micro-Tom expressing Cas9. Second, we devised a transgene-free GE method based on a potato virus X (PVX) vector to deliver Cas9 and sgRNAs. We designed and cloned sgRNAs targeting Phytoene desaturase in the VIGE vectors and determined optimal conditions for VIGE. We evaluated VIGE efficiency through deep sequencing of the target gene after viral vector inoculation, detecting 40.3% and 36.5% mutation rates for TRV- and PVX-mediated GE, respectively. To improve editing efficiency, we applied a 37°C heat treatment, which increased the editing efficiency by 33–46% and 56–76% for TRV- and PVX-mediated VIGE, respectively. To obtain edited plants, we subjected inoculated cotyledons to tissue culture, yielding successful editing events. We also demonstrated that PVX-mediated GE can be applied to other Solanaceous crops, such as potato (Solanum tuberosum) and eggplant (Solanum melongena). These simple and highly efficient VIGE methods have great potential for generating genome-edited plants in Solanaceous crops.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of virus-induced genome editing methods in Solanaceous crops

Lee,

Kang,

Venkatesh

et al. 2023

Horticulture Research

View full text Add to dashboard Cite

show abstract

“…Згідно з опублікованими даними, більшість трансгенних рослин пшениці отримані за допомогою прямого переносу генів -біолістичним методом (Altpeter et al, 1996;Lazzeri, Jones 2009;Vasil et al, 1992;Weeks et al, 1993;Zhou et al, 1995;Qin et al, 2014;Hamada et al, 2017). Тим не менш, така трансформація має ряд обмежень та недоліків, основним з яких є можливі перебудови перенесених ДНК касет та мультикопійність, що зумовлюють «замовчування» відповідних генів (Бончук и др., 2014: пат.…”

unclassified

Study of transgene expression in Triticum aestivum L. after Agrobacterium-mediated in planta transformation

Zhalii¹,

Bannikova²,

Plugatar³

et al. 2019

Visn. ukr. tov. genet. sel.

View full text Add to dashboard Cite

Aim. Detection of sequences of target transgenes nptII and bar in the genome of probable transformants of bread winter wheat Triticum aestivum L. cultivars Zymoiarka and Podolianka obtained as a result of Agrobacterium-mediated transformation in planta and determination of their expression level. Methods. Polymerase chain reaction (PCR) method was used independently and in combination with reverse transcription (RT-PCR), electrophoresis of DNA in agarose gel. Tolerance to the herbicide was evaluated in the physiological test. Results Through PCR analysis, the sequence of nptII transgene was detected in 30 samples of 145 analyzed, the frequency of transformation was 20.7 %. The sequence of the gene bar was observed in 85 experimental plants, and the frequency of transformation was 15.6 %. mRNAs of both transgenes were detected, indicating their transcriptional activity and stable expression. Conclusions PCR analysis allowed to detect nptII transgenic signal in 20.7 % of plants, while the presence of the bar gene was detected in 15.6 % of cases, indicating a higher efficiency of this genetic construct. The transcription is shown in all the specimens studied for both transgene. According to the results of the physiological test, 25 % of plants containing the gene bar showed resistance to the Basta® herbicide.Keywords: genetically modified organisms, transgenic plants, biotechnological cultures, bread winter wheat, genetic engineering.

show abstract