scholarly journals CRISPR/Cas9-mediated efficient and heritable targeted mutagenesis in tomato plants in the first and later generations

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Changtian Pan ◽  
Lei Ye ◽  
Li Qin ◽  
Xue Liu ◽  
Yanjun He ◽  
...  
Keyword(s):  
Average Frequency ◽  
Transformation Method ◽  
Targeted Mutagenesis ◽  
Efficient Tool ◽  
Tomato Plants ◽  
High Frequencies ◽  
High Mutation Frequency ◽  
Transgenic Tomato Plants ◽  
Targeted Gene Editing ◽  
The Stability

Abstract The CRISPR/Cas9 system has successfully been used in various organisms for precise targeted gene editing. Although it has been demonstrated that CRISPR/Cas9 system can induce mutation in tomato plants, the stability of heredity in later generations and mutant specificity induced by the CRISPR/Cas9 system in tomato plants have not yet been elucidated in detail. In this study, two genes, SlPDS and SlPIF4, were used for testing targeted mutagenesis in tomato plants through an Agrobacterium tumefaciens-mediated transformation method. A high mutation frequency was observed in all tested targets in the T0 transgenic tomato plants, with an average frequency of 83.56%. Clear albino phenotypes were observed for the psd mutants. High frequencies of homozygous and biallelic mutants were detected even in T0 plants. The majority of the detected mutations were 1- to 3-nucleotide deletions, followed by 1-bp insertions. The target mutations in the T0 lines were stably transmitted to the T1 and T2 generations, without new modifications or revision. Off-target activities associated with SlPDS and SlPIF4 were also evaluated by sequencing the putative off-target sites and no clear off-target events were detected. Our results demonstrate that the CRISPR/Cas9 system is an efficient tool for generating stable and heritable modifications in tomato plants.

scholarly journals Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System

2018 ◽  
Vol 19 (10) ◽  
pp. 3000 ◽  
Author(s):  
Shouhong Zhu ◽  
Xiuli Yu ◽  
Yanjun Li ◽  
Yuqiang Sun ◽  
Qianhao Zhu ◽  
...  
Keyword(s):  
Gene Editing ◽  
Crop Improvement ◽  
Targeted Mutagenesis ◽  
Cotton Fibers ◽  
Efficient Tool ◽  
Gene Encoding ◽  
Editing Event ◽  
Highly Efficient ◽  
Target Sites ◽  
Targeted Gene Editing

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system has been shown to be able to induce highly efficient mutagenesis in the targeted DNA of many plants, including cotton, and has become an important tool for investigation of gene function and crop improvement. Here, we developed a simple and easy to operate CRISPR/Cas9 system and demonstrated its high editing efficiency in cotton by targeting-ALARP, a gene encoding alanine-rich protein that is preferentially expressed in cotton fibers. Based on sequence analysis of the target site in the 10 transgenic cottons containing CRISPR/Cas9, we found that the mutation frequencies of GhALARP-A and GhALARP-D target sites were 71.4–100% and 92.9–100%, respectively. The most common editing event was deletion, but deletion together with large insertion was also observed. Mosaic mutation editing events were detected in most transgenic plants. No off-target mutation event was detected in any the 15 predicted sites analyzed. This study provided mutants for further study of the function of GhALARP in cotton fiber development. Our results further demonstrated the feasibility of use of CRISPR/Cas9 as a targeted mutagenesis tool in cotton, and provided an efficient tool for targeted mutagenesis and functional genomics in cotton.

scholarly journals Overexpression of Pear (Pyrus pyrifolia) CAD2 in Tomato Affects Lignin Content

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2595 ◽  
Author(s):  
Mingtong Li ◽  
Chenxia Cheng ◽  
Xinfu Zhang ◽  
Suping Zhou ◽  
Lixia Li ◽  
...  
Keyword(s):  
Lignin Content ◽  
Lignin Biosynthesis ◽  
Cinnamyl Alcohol Dehydrogenase ◽  
Transformation Method ◽  
Transgenic Tomato ◽  
Pyrus Pyrifolia ◽  
Tomato Plants ◽  
Over Expression ◽  
Transgenic Tomato Plants ◽  
Vessel Elements

PpCAD2 was originally isolated from the ‘Wangkumbae’ pear (Pyrus pyrifolia Nakai), and it encodes for cinnamyl alcohol dehydrogenase (CAD), which is a key enzyme in the lignin biosynthesis pathway. In order to verify the function of PpCAD2, transgenic tomato (Solanum lycopersicum) ‘Micro-Tom’ plants were generated using over-expression constructs via the agrobacterium-mediated transformation method. The results showed that the PpCAD2 over-expression transgenic tomato plant had a strong growth vigor. Furthermore, these PpCAD2 over-expression transgenic tomato plants contained a higher lignin content and CAD enzymatic activity in the stem, leaf and fruit pericarp tissues, and formed a greater number of vessel elements in the stem and leaf vein, compared to wild type tomato plants. This study clearly indicated that overexpressing PpCAD2 increased the lignin deposition of transgenic tomato plants, and thus validated the function of PpCAD2 in lignin biosynthesis.

scholarly journals Generation and molecular characterization of CRISPR/Cas9-induced mutations in 63 immunity-associated genes in tomato reveals specificity and a range of gene modifications

2019 ◽  
Author(s):  
Ning Zhang ◽  
Holly M. Roberts ◽  
Joyce Van Eck ◽  
Gregory B. Martin
Keyword(s):  
Molecular Characterization ◽  
Gene Editing ◽  
Induced Mutations ◽  
Tomato Plants ◽  
Transgenic Tomato Plants ◽  
Target Sites ◽  
Targeted Gene Editing ◽  
Gene Alterations ◽  
New Mutations

AbstractThe CRISPR/Cas9 system is a powerful tool for targeted gene editing in many organisms including plants. However, most of the reported uses of CRISPR/Cas9 in plants have focused on modifying one or a few genes, and thus the overall specificity, types of mutations, and heritability of gene alterations remain unclear. Here we describe the molecular characterization of 361 T0 transgenic tomato plants that were generated using CRISPR/Cas9 to induce mutations in 63 immunity-associated genes. Among the T0 transformed plants, 245 carried mutations (68%), with 20% of those plants being homozygous for the mutation, 30% being heterozygous, 32% having two different mutations (biallelic) and 18% having multiple mutations (chimeric). The mutations were predominantly short insertions or deletions, with 87% of the affected sequences being smaller than 10 bp. The majority of 1 bp insertions were A (50%) or T (29%). The mutations from the T0 generation were stably transmitted to later generations, although new mutations were detected in some T1 plants. No mutations were detected in 18 potential off-target sites among 144 plants. Our study provides a broad and detailed view into the effectiveness of CRISPR/Cas9 for genome editing in an economically important plant species.

Dark respiration rate of transgenic tomato plants expressing FeSOD1 gene under chloride and sulfate salinity

2014 ◽  
Vol 40 (1) ◽  
pp. 14-17 ◽  
Author(s):  
Ye. N. Baranova ◽  
E. N. Akanov ◽  
A. A. Gulevich ◽  
L. V. Kurenina ◽  
S. A. Danilova ◽  
...  
Keyword(s):  
Respiration Rate ◽  
Dark Respiration ◽  
Transgenic Tomato ◽  
Dark Respiration Rate ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

Genetic transactivation of Dissociation elements in transgenic tomato plants

1989 ◽  
Vol 218 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Michael W. Lassner ◽  
Joseph M. Palys ◽  
John I. Yoder
Keyword(s):  
Transgenic Tomato ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

Overexpression of LeNHX2 and SlSOS2 increases salt tolerance and fruit production in double transgenic tomato plants

2019 ◽  
Vol 135 ◽  
pp. 77-86 ◽  
Author(s):  
Mourad Baghour ◽  
Francisco Javier Gálvez ◽  
M. Elena Sánchez ◽  
M. Nieves Aranda ◽  
Kees Venema ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Fruit Production ◽  
Transgenic Tomato ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

scholarly journals Control of Ethylene Synthesis by Expression of a Bacterial Enzyme in Transgenic Tomato Plants

1991 ◽  
Vol 3 (11) ◽  
pp. 1187 ◽  
Author(s):  
Harry J. Klee ◽  
Maria B. Hayford ◽  
Keith A. Kretzmer ◽  
Gerard F. Barry ◽  
Ganesh M. Kishore
Keyword(s):  
Transgenic Tomato ◽  
Ethylene Synthesis ◽  
Tomato Plants ◽  
Bacterial Enzyme ◽  
Transgenic Tomato Plants

Amplifier Performance in Terms of Two-Port Parameters

1998 ◽  
Vol 35 (1) ◽  
pp. 47-70
Author(s):  
Jack Willis
Keyword(s):  
Feedback Loop ◽  
High Frequencies ◽  
Feedback Amplifier ◽  
Alternative Means ◽  
The Stability

At high frequencies the complexity of the circuit representing a feedback amplifier is such that the concept of a feedback loop becomes meaningless, and an alternative means of checking the stability of the amplifier is needed. Representing the network as a two-port realises such a solution.

Discrete-Time H-Infinity Synthesis of Frequency-Shaped Sliding Mode Control for Suppression of Vibration With Multiple Peak Frequencies

2016 ◽  
Author(s):  
Minghui Zheng ◽  
Masayoshi Tomizuka
Keyword(s):  
High Frequency ◽  
Sliding Mode ◽  
Disk Drive ◽  
Sliding Surface ◽  
Surface Dynamics ◽  
H Infinity ◽  
High Frequencies ◽  
Multiple Peak ◽  
Significant Performance ◽  
The Stability

Vibration with multiple large peaks at high frequencies may cause significant performance degradation and have become a major concern in modern high precision control systems. To deal with such high-frequency peaks, it is proposed to design a frequency-shaped sliding mode controller based on H∞ synthesis. It obtains an ‘optimal’ filter to shape the sliding surface, and thus provides frequency-dependent control allocation. The proposed frequency-shaping method assures the stability in the presence of multiple-peak vibration sources, and minimizes the weighted H∞ norm of the sliding surface dynamics. The evaluation is performed on a simulated hard disk drive with actual vibration sources from experiments, and the effectiveness of large vibration peak suppression is demonstrated.

scholarly journals Construction of CRISPR/Cas9 expression vectors habouring gRNA targeted on SLIAA9 gene of tomato

2020 ◽  
Vol 18 (1) ◽  
pp. 147-156
Author(s):  
Bui Manh Minh ◽  
Ha Hong Hanh ◽  
Le Thi Thu Hien ◽  
Huynh Thi Thu Hue
Keyword(s):  
Transgenic Plants ◽  
Genome Editing ◽  
Tomato Fruit ◽  
Expression System ◽  
Secondary Compounds ◽  
Expression Vectors ◽  
Tomato Plants ◽  
Normal Morphology ◽  
Transgenic Tomato Plants ◽  
Pcr Test

Tomato (Solanum lycopersicum) is a nutritious fruit containing many secondary compounds with health benefits. The formation of tomato fruit through fertilization is controlled by auxin through Aux/IAA9 and ARF8 proteins. The mutated SlIAA9 gene leads to the parthenocarpic development of fruit or seedless tomato fruit. Nowadays, the CRISPR/Cas9 genome editing system is becoming increasingly popular in modifying desired genes on plant objects. In this study, gRNAs which target on tomato SlIAA9 gene were designed and inserted into CRISPR/Cas9 vectors. In addition, two strains of A. tumefaciens harboring pRGEB31-IAA9G2 and pRGEB32-IAA9G2 vectors carrying CRISPR/Cas9 expression system towards SlIAA9 gene in tomato were successfully created. The strain of A. tumefaciens harboring pRGEB31- IAA9G2 plasmid was used to develop transgenic tomato plants from Micro-Tom variety. PCR test showed that 5/14 plants had the presence of Cas9 gene in T0 plants. The transgenic plants have a normal morphology in comparation with the controls. The evaluation of mutant efficiency, type, and stability of mutations on the SlIAA9 will be conducted on next-generation plants when the mutations are stable and segregated into descendents.

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