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https://www.readbyqxmd.com/read/28913803/doubled-haploid-transgenic-wheat-lines-by-microspore-transformation
#1
Sachin Rustgi, Nii O Ankrah, Rhoda A T Brew-Appiah, Yue Sun, Weiguo Liu, Diter von Wettstein
Microspores are preferred explant choice for genetic transformation, as their use shortens the duration of obtaining homozygous transformants. All established gene-delivery methods of particle bombardment, electroporation, and cocultivation with Agrobacterium tumefaciens were optimized on androgenic microspores or derived tissues. In the biolistic gene delivery method 35-40 days old haploid microspore embryoids were used for genetic transformation, whereas freshly isolated androgenic microspores were used for genetic transformation in the electroporation and Agrobacterium cocultivation-based methods...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28913799/biolistic-transformation-of-wheat
#2
Caroline Tassy, Pierre Barret
The wheat genome encodes some 100,000 genes. To understand how the expression of these genes is regulated it will be necessary to carry out many genetic transformation experiments. Robust protocols that allow scientists to transform a wide range of wheat genotypes are therefore required. In this chapter, we describe a protocol for biolistic transformation of wheat that uses immature embryos and small quantities of DNA cassettes. An original method for DNA cassette purification is also described. This protocol can be used to transform a wide range of wheat genotypes and other related species...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28904403/an-in-planta-biolistic-method-for-stable-wheat-transformation
#3
Haruyasu Hamada, Qianyan Linghu, Yozo Nagira, Ryuji Miki, Naoaki Taoka, Ryozo Imai
The currently favoured method for wheat (Triticum aestivum L.) transformation is inapplicable to many elite cultivars because it requires callus culture and regeneration. Here, we developed a simple, reproducible, in planta wheat transformation method using biolistic DNA delivery without callus culture or regeneration. Shoot apical meristems (SAMs) grown from dry imbibed seeds were exposed under a microscope and subjected to bombardment with different-sized gold particles coated with the GFP gene construct, introducing DNA into the L2 cell layer...
September 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28861822/transient-expression-of-chimeric-fluorescent-reporter-proteins-in-pollen-tubes-to-study-protein-polar-secretion-and-dynamics
#4
Guitao Zhong, Ronghe Liu, Menglong Zhuang, Hao Wang
Transient expression of chimeric fluorescent reporter proteins by biolistic bombardment is a quick and useful procedure for studying subcellular protein localization and dynamics in plants. It is especially beneficial in specific plant cells which are not suitable for protoplast-based and Agrobacterium-mediated protein transient expression. Polar protein secretion and vesicular trafficking play essential functions for cell polarization and tip growth. The growing pollen tube is regarded as an ideal model plant cell system to study the machinery and regulation of polar protein trafficking and targeting...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28815500/nanoparticle-mediated-recombinase-delivery-into-maize
#5
Susana Martin-Ortigosa, Brian G Trewyn, Kan Wang
We describe a non-DNA-based system for delivering Cre recombinase protein into maize tissue using gold-plated mesoporous silica nanoparticle (Au-MSN). Cre protein is first loaded into the pores of Au-MSNs and then delivered using the biolistic method to immature embryos of a maize line (Lox-corn), which harbors loxP sites flanking a selection and a reporter gene. The release of the Cre recombinase protein inside the plant cell leads to recombination at the loxP sites, eliminating both genes. Visual screening is used to identify recombination events, which can be regenerated to mature and fertile plants...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28791544/infectivity-of-deinbollia-mosaic-virus-a-novel-weed-infecting-begomovirus-in-east-africa
#6
Martina Kyallo, Elijah Miinda Ateka, Peter Sseruwagi, José Trinidad Ascencio-Ibáñez, Mildred-Ochwo Ssemakula, Robert Skilton, Joseph Ndunguru
Weed-infecting begomoviruses play an important role in the epidemiology of crop diseases because they can potentially infect crops and contribute to the genetic diversity of crop-infecting begomoviruses. Despite the important epidemiological role that weed-infecting begomoviruses play, they remain insufficiently studied in Africa. Recently, we identified Deinbollia mosaic virus (DMV), a distinct begomovirus found naturally infecting the weed host Deinbollia borbonica (Sapindaceae) in Kenya and Tanzania. In this study, we investigated the capacity of DMV to infect a restricted host range of Solanaceae and Euphorbiaceae species...
August 9, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28667403/molecular-and-fish-analyses-of-a-53-kbp-intact-dna-fragment-inserted-by-biolistics-in-wheat-triticum-aestivum-l-genome
#7
A Partier, G Gay, C Tassy, M Beckert, C Feuillet, P Barret
A large, 53-kbp, intact DNA fragment was inserted into the wheat ( Triticum aestivum L.) genome. FISH analyses of individual transgenic events revealed multiple insertions of intact fragments. Transferring large intact DNA fragments containing clusters of resistance genes or complete metabolic pathways into the wheat genome remains a challenge. In a previous work, we showed that the use of dephosphorylated cassettes for wheat transformation enabled the production of simple integration patterns. Here, we used the same technology to produce a cassette containing a 44-kb Arabidopsis thaliana BAC, flanked by one selection gene and one reporter gene...
June 30, 2017: Plant Cell Reports
https://www.readbyqxmd.com/read/28539970/developing-transgenic-wheat-to-encounter-rusts-and-powdery-mildew-by-overexpressing-barley-chi26-gene-for-fungal-resistance
#8
Hala F Eissa, Sameh E Hassanien, Ahmed M Ramadan, Moustafa M El-Shamy, Osama M Saleh, Ahmed M Shokry, Mohamed Abdelsattar, Yasser B Morsy, Maher A El-Maghraby, Hussien F Alameldin, Sabah M Hassan, Gamal H Osman, Hesham T Mahfouz, Gharib A Gad El-Karim, Magdy A Madkour, Ahmed Bahieldin
BACKGROUND: The main aim of this study was to improve fungal resistance in bread wheat via transgenesis. Transgenic wheat plants harboring barley chitinase (chi26) gene, driven by maize ubi promoter, were obtained using biolistic bombardment, whereas the herbicide resistance gene, bar, driven by the CaMV 35S promoter was used as a selectable marker. RESULTS: Molecular analysis confirmed the integration, copy number, and the level of expression of the chi26 gene in four independent transgenic events...
2017: Plant Methods
https://www.readbyqxmd.com/read/28395198/propagation-and-some-physiological-effects-of-citrus-bark-cracking-viroid-and-apple-fruit-crinkle-viroid-in-multiple-infected-hop-humulus-lupulus-l
#9
J Matoušek, K Siglová, J Jakše, S Radišek, Joseph R J Brass, T Tsushima, T Guček, G S Duraisamy, T Sano, G Steger
The hop metabolome important for the brewing industry and for medical purposes is endangered worldwide due to multiple viroid infections affecting hop physiology. Combinatorial biolistic hop inoculation with Citrus bark cracking viroid (CBCVd), Apple fruit crinkle viroid (AFCVd), Hop latent viroid, and Hop stunt viroid (HSVd) showed a low CBCVd compatibility with HSVd, while all other viroid combinations were highly compatible. Unlike to other viroids, single CBCVd propagation showed a significant excess of (-) over (+) strands in hop, tomato, and Nicotiana benthamiana, but not in citruses...
June 2017: Journal of Plant Physiology
https://www.readbyqxmd.com/read/28274738/labeling-of-neuronal-morphology-using-custom-diolistic-techniques
#10
Lyon H Hough, Michael E Brown
BACKGROUND: Diolistic labeling is increasingly utilized in neuroscience as an efficient, reproducible method for visualization of neuronal morphology. The use of lipophilic carbocyanine dyes, combined with particle-mediated biolistic delivery allows for non-toxic fluorescent labeling of multiple neurons in both living and fixed tissue. Since first described, this labeling method has been modified to fit a variety of research goals and laboratory settings. NEW METHOD: Diolistic labeling has traditionally relied on commercially available devices for the propulsion of coated micro-particles into tissue sections...
March 6, 2017: Journal of Neuroscience Methods
https://www.readbyqxmd.com/read/28270156/a-rapid-virus-induced-gene-silencing-vigs-method-for-assessing-resistance-and-susceptibility-to-cassava-mosaic-disease
#11
Getu Beyene, Raj Deepika Chauhan, Nigel J Taylor
BACKGROUND: Cassava mosaic disease (CMD) is a major constraint to cassava production in sub-Saharan Africa. Under field conditions, evaluation for resistance to CMD takes 12-18 months, often conducted across multiple years and locations under pressure from whitefly-mediated transmission. Under greenhouse or laboratory settings, evaluation for resistance or susceptibility to CMD involves transmission of the causal viruses from an infected source to healthy plants through grafting, or by using Agrobacterium-mediated or biolistic delivery of infectious clones...
March 7, 2017: Virology Journal
https://www.readbyqxmd.com/read/28148855/viral-vector-based-transduction-of-slice-cultures
#12
J Simon Wiegert, Christine E Gee, Thomas G Oertner
Transgenes can be introduced into the cells of organotypic slice cultures using different delivery methods, such as biolistic transfection, electroporation, and viral vector-based transduction. These methods produce different patterns of transgene expression. Local injection of recombinant adeno-associated virus (rAAV) produces a small cluster of transgene-expressing neurons around the injection site. Expression in individual cells varies with the distance from the injection site, indicating that many neurons take up several rAAV particles...
February 1, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28132145/agrobacterium-mediated-transformation-of-leaf-base-segments
#13
Sebastian Gasparis
Agrobacterium-mediated transformation has become a routine method of genetic engineering of cereals, gradually replacing the biolistic protocols. Simple integration patterns of transgenic loci, decent transformation efficiency, and technical simplicity are the main advantages offered by this method. Here we present a detailed protocol for the production of transgenic oat plants by Agrobacterium-mediated transformation of leaf base segments. The use of leaf explants as target tissues for transformation and in vitro regeneration of transgenic plants may be a good alternative for genotypes which are not susceptible to regeneration from immature or mature embryos...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28120101/virus-induced-gene-silencing-in-rauwolfia-species
#14
Cyrielle Corbin, Florent Lafontaine, Liuda Johana Sepúlveda, Ines Carqueijeiro, Martine Courtois, Arnaud Lanoue, Thomas Dugé de Bernonville, Sébastien Besseau, Gaëlle Glévarec, Nicolas Papon, Lucia Atehortúa, Nathalie Giglioli-Guivarc'h, Marc Clastre, Benoit St-Pierre, Audrey Oudin, Vincent Courdavault
Elucidation of the monoterpene indole alkaloid biosynthesis has recently progressed in Apocynaceae through the concomitant development of transcriptomic analyses and reverse genetic approaches performed by virus-induced gene silencing (VIGS). While most of these tools have been primarily adapted for the Madagascar periwinkle (Catharanthus roseus), the VIGS procedure has scarcely been used on other Apocynaceae species. For instance, Rauwolfia sp. constitutes a unique source of specific and valuable monoterpene indole alkaloids such as the hypertensive reserpine but are also well recognized models for studying alkaloid metabolism, and as such would benefit from an efficient VIGS procedure...
July 2017: Protoplasma
https://www.readbyqxmd.com/read/28081289/transgenic-cotton-expressing-cry10aa-toxin-confers-high-resistance-to-the-cotton-boll-weevil
#15
Thuanne Pires Ribeiro, Fabricio Barbosa Monteiro Arraes, Isabela Tristan Lourenço-Tessutti, Marilia Santos Silva, Maria Eugênia Lisei-de-Sá, Wagner Alexandre Lucena, Leonardo Lima Pepino Macedo, Janaina Nascimento Lima, Regina Maria Santos Amorim, Sinara Artico, Márcio Alves-Ferreira, Maria Cristina Mattar Silva, Maria Fatima Grossi-de-Sa
Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation...
August 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28071976/precise-flexible-and-affordable-gene-stacking-for-crop-improvement
#16
Weiqiang Chen, David W Ow
The genetic engineering of plants offers a revolutionary advance for crop improvement, and the incorporation of transgenes into crop species can impart new traits that would otherwise be difficult to obtain through conventional breeding. Transgenes introduced into plants, however, can only be useful when bred out to field cultivars. As new traits are continually added to further improve transgenic cultivars, clustering new DNA near previously introduced transgenes keep from inflating the number of segregating units that breeders must assemble back into a breeding line...
January 10, 2017: Bioengineered
https://www.readbyqxmd.com/read/28005227/in-planta-production-and-characterization-of-a-hyperthermostable-gh10-xylanase-in-transgenic-sugarcane
#17
Jae Yoon Kim, Guang Nong, John D Rice, Maria Gallo, James F Preston, Fredy Altpeter
Sugarcane (Saccharum sp. hybrids) is one of the most efficient and sustainable feedstocks for commercial production of fuel ethanol. Recent efforts focus on the integration of first and second generation bioethanol conversion technologies for sugarcane to increase biofuel yields. This integrated process will utilize both the cell wall bound sugars of the abundant lignocellulosic sugarcane residues in addition to the sucrose from stem internodes. Enzymatic hydrolysis of lignocellulosic biomass into its component sugars requires significant amounts of cell wall degrading enzymes...
March 2017: Plant Molecular Biology
https://www.readbyqxmd.com/read/27973750/controlled-release-and-intracellular-protein-delivery-from-mesoporous-silica-nanoparticles
#18
REVIEW
Gauri V Deodhar, Marisa L Adams, Brian G Trewyn
Protein therapeutics are promising candidates for disease treatment due to their high specificity and minimal adverse side effects; however, targeted protein delivery to specific sites has proven challenging. Mesoporous silica nanoparticles (MSN) have demonstrated to be ideal candidates for this application, given their high loading capacity, biocompatibility, and ability to protect host molecules from degradation. These materials exhibit tunable pore sizes, shapes and volumes, and surfaces which can be easily functionalized...
January 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/27943461/high-efficiency-gene-targeting-in-hexaploid-wheat-using-dna-replicons-and-crispr-cas9
#19
Javier Gil-Humanes, Yanpeng Wang, Zhen Liang, Qiwei Shan, Carmen V Ozuna, Susana Sánchez-León, Nicholas J Baltes, Colby Starker, Francisco Barro, Caixia Gao, Daniel F Voytas
The ability to edit plant genomes through gene targeting (GT) requires efficient methods to deliver both sequence-specific nucleases (SSNs) and repair templates to plant cells. This is typically achieved using Agrobacterium T-DNA, biolistics or by stably integrating nuclease-encoding cassettes and repair templates into the plant genome. In dicotyledonous plants, such as Nicotinana tabacum (tobacco) and Solanum lycopersicum (tomato), greater than 10-fold enhancements in GT frequencies have been achieved using DNA virus-based replicons...
March 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/27848933/genome-editing-in-maize-directed-by-crispr-cas9-ribonucleoprotein-complexes
#20
Sergei Svitashev, Christine Schwartz, Brian Lenderts, Joshua K Young, A Mark Cigan
Targeted DNA double-strand breaks have been shown to significantly increase the frequency and precision of genome editing. In the past two decades, several double-strand break technologies have been developed. CRISPR-Cas9 has quickly become the technology of choice for genome editing due to its simplicity, efficiency and versatility. Currently, genome editing in plants primarily relies on delivering double-strand break reagents in the form of DNA vectors. Here we report biolistic delivery of pre-assembled Cas9-gRNA ribonucleoproteins into maize embryo cells and regeneration of plants with both mutated and edited alleles...
November 16, 2016: Nature Communications
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