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Plant Methods

Israel A Ramírez-Alanis, Justin B Renaud, Silverio García-Lara, Rima Menassa, Guy A Cardineau
Background: Expression of economically relevant proteins in alternative expression platforms, especially plant expression platforms, has gained significant interest in recent years. A special interest in working with plants as bioreactors for the production of pharmaceutical proteins is related to low production costs, product safety and quality. Among the different properties that plants can also offer for the production of recombinant proteins, protein glycosylation is crucial since it may have an impact on pharmaceutical functionality and/or stability...
2018: Plant Methods
Iván Jauregui, Shane A Rothwell, Samuel H Taylor, Martin A J Parry, Elizabete Carmo-Silva, Ian C Dodd
Background: Improving plant water use efficiency (WUE) is a major target for improving crop yield resilience to adverse climate change. Identifying genetic variation in WUE usually relies on instantaneous measurements of photosynthesis (An) and transpiration (Tr), or integrative measurements of carbon isotope discrimination, at the leaf level. However, leaf gas exchange measurements alone do not adequately represent whole plant responses, especially if evaporative demand around the plant changes...
2018: Plant Methods
Charlotte Bureau, Nadège Lanau, Mathieu Ingouff, Boukhaddaoui Hassan, Anne-Cécile Meunier, Fanchon Divol, Rosie Sevilla, Delphine Mieulet, Anne Dievart, Christophe Périn
Background: The clear visualization of 3D organization at the cellular level in plant tissues is needed to fully understand plant development processes. Imaging tools allow the visualization of the main fluorophores and in vivo growth monitoring. Confocal microscopy coupled with the use of propidium iodide (PI) counter-staining is one of the most popular tools used to characterize the structure of root meristems in A. thaliana . However, such an approach is relatively ineffective in species with more complex and thicker root systems...
2018: Plant Methods
S V G N Priyadarshani, Bingyan Hu, Weimin Li, Hina Ali, Haifeng Jia, Lihua Zhao, Simon Peter Ojolo, Syed Muhammad Azam, Junjie Xiong, Moakai Yan, Zia Ur Rahman, Qingsong Wu, Yuan Qin
Background: An efficient transformation protocol is a primary requisite to study and utilize the genetic potential of any plant species. A quick transformation system is also crucial for the functional analysis of genes along with the study of proteins and their interactions in vivo. Presently, however, quick and effective transformation systems are still lacking for many plant species including pineapple. This has limited the full exploration of the genetic repository of pineapple as well as the study of its genes, protein localization and protein interactions...
2018: Plant Methods
Hector Camargo Alvarez, Melba Salazar-Gutiérrez, Diana Zapata, Markus Keller, Gerrit Hoogenboom
Background: The reduced growth of plants during the winter causes a lack in the perceptibility of the phenological events making challenging the study of dormancy. For deciduous crops, dormancy is generally determined by evaluating budbreak of single-node cuttings that are exposed to conditions suitable for growth. However, the absence of a statistical basis for analyzing and interpreting the budbreak behavior evaluated as the percent budbreak, the average time to budbreak and the time to reach 50% budbreak, has caused inconsistent and contradictory criteria to identify the dormancy status of different deciduous crops...
2018: Plant Methods
Sadiye Hayta, Mark A Smedley, Jinhong Li, Wendy A Harwood, Philip M Gilmartin
Background: Genetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics. Primulaceae species provide one of the best-known examples of heteromorphic flower development, a breeding system which has attracted considerable attention, including that of Charles Darwin. Molecular approaches, including plant transformation give the best opportunity to define and understand the role of genes involved in floral heteromorphy in the common primrose, Primula vulgaris , along with other Primula species...
2018: Plant Methods
Catalina Salgado-Salazar, Gary R Bauchan, Emma C Wallace, Jo Anne Crouch
Background: Plasmopara obducens is the biotrophic oomycete responsible for impatiens downy mildew, a destructive disease of Impatiens that causes high crop loss. Currently, there are no available methods for the microscopic detection of P. obducens from leaves of impatiens, which may be contributing to the spread of the disease. Fluorescence in situ hybridization (FISH) is a sensitive and robust method that uses sequence-specific, fluorescence-labeled oligonucleotide probes to detect target organisms from the environment...
2018: Plant Methods
Michael A Reeve, Alan G Buddie
Background: Protein-containing samples can readily be characterised and/or identified using matrix-assisted laser-desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS). This technique however requires relatively-fresh biological material that contains proteins that have not yet undergone significant degradation. For field-work collection of samples, problems are often encountered due to delays between collection and sample processing, sample storage (possibly at elevated temperature and/or humidity in some climates), quarantine/regulatory restrictions on the transfer of living biological materials across national borders, and the potential to transfer unwanted microorganisms via non-living biological materials...
2018: Plant Methods
Matthew D Fischer, Emmanuel Mgboji, Zhongchi Liu
Background: Insertion of engineered DNA fragments into bacterial vectors is the foundation of recombinant DNA technology, yet existing methods are still laborious, require many steps, depend on specific vector configuration, or require expensive reagents. Results: We have developed a method, called "Pyrite" cloning that combines the traditional restriction enzyme digestion and ligation reaction in a single tube and uses a programmed thermocycler reaction, allowing rapid and flexible cloning in a single tube...
2018: Plant Methods
Kevin Vidot, Cédric Gaillard, Camille Rivard, René Siret, Marc Lahaye
Background: The in vivo observation of diffusible components, such as ions and small phenolic compounds, remains a challenge in turgid plant organs. The analytical techniques used to localize such components in water-rich tissue with a large field of view are lacking. It remains an issue to limit compound diffusion during sample preparation and observation processes. Results: An experimental setup involving the infusion staining of plant tissue and the cryo-fixation and cryo-sectioning of tissue samples followed by fluorescence cryo-observation by laser scanning confocal microscopy (LSCM) was developed...
2018: Plant Methods
Lidiane Lindinalva Barbosa Amorim, José Ribamar Costa Ferreira-Neto, João Pacífico Bezerra-Neto, Valesca Pandolfi, Flávia Tadeu de Araújo, Mitalle Karen da Silva Matos, Mauro Guida Santos, Ederson Akio Kido, Ana Maria Benko-Iseppon
Background: Due to cowpea ability to fix nitrogen in poor soils and relative tolerance to drought and salt stresses, efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Real-time quantitative PCR (qPCR) has been widely used as the most reliable method to measure gene expression, due to its high accuracy and specificity. In the present study, nine candidate reference genes were rigorously tested for their application in normalization of qPCR data onto roots of four distinct cowpea accessions under two abiotic stresses: root dehydration and salt (NaCl, 100 mM)...
2018: Plant Methods
Min-Rui Wang, Zhen-Hua Cui, Jing-Wei Li, Xin-Yi Hao, Lei Zhao, Qiao-Chun Wang
Production of virus-free plants is necessary to control viral diseases, import novel cultivars from other countries, exchange breeding materials between countries or regions and preserve plant germplasm. In vitro techniques represent the most successful approaches for virus eradication. In vitro thermotherapy-based methods, including combining thermotherapy with shoot tip culture, chemotherapy, micrografting or shoot tip cryotherapy, have been successfully established for efficient eradication of various viruses from almost all of the most economically important crops...
2018: Plant Methods
Koushik Nagasubramanian, Sarah Jones, Soumik Sarkar, Asheesh K Singh, Arti Singh, Baskar Ganapathysubramanian
Background: Charcoal rot is a fungal disease that thrives in warm dry conditions and affects the yield of soybeans and other important agronomic crops worldwide. There is a need for robust, automatic and consistent early detection and quantification of disease symptoms which are important in breeding programs for the development of improved cultivars and in crop production for the implementation of disease control measures for yield protection. Current methods of plant disease phenotyping are predominantly visual and hence are slow and prone to human error and variation...
2018: Plant Methods
Lu Long, Dan-Dan Guo, Wei Gao, Wen-Wen Yang, Li-Pan Hou, Xiao-Nan Ma, Yu-Chen Miao, Jose Ramon Botella, Chun-Peng Song
Background: When developing CRISPR/Cas9 systems for crops, it is crucial to invest time characterizing the genome editing efficiency of the CRISPR/Cas9 cassettes, especially if the transformation system is difficult or time-consuming. Cotton is an important crop for the production of fiber, oil, and biofuel. However, the cotton stable transformation is usually performed using Agrobacterium tumefaciens taking between 8 and 12 months to generate T0 plants. Furthermore, cotton is a heterotetraploid and targeted mutagenesis is considered to be difficult as many genes are duplicated in this complex genome...
2018: Plant Methods
Gernot Bodner, Alireza Nakhforoosh, Thomas Arnold, Daniel Leitner
Background: Root phenotyping aims to characterize root system architecture because of its functional role in resource acquisition. RGB imaging and analysis procedures measure root system traits via colour contrasts between roots and growth media or artificial backgrounds. In the case of plants grown in soil-filled rhizoboxes, where the colour contrast can be poor, it is hypothesized that root imaging based on spectral signatures improves segmentation and provides additional knowledge on physico-chemical root properties...
2018: Plant Methods
Umidjon Shapulatov, Mark van Hoogdalem, Marielle Schreuder, Harro Bouwmeester, Ibrokhim Y Abdurakhmonov, Alexander R van der Krol
Background: Recently, putative pre-miRNAs locations have been identified in the introns of plant genes, raising the question whether such genes can show a dual functionality by having both correct maturation of the host gene pre-mRNA and maturation of the miRNAs from the intron. Here, we demonstrated that such dual functionality is indeed possible, using as host gene the firefly luciferase gene with intron (ffgLUC), and different artificial intronic miRNAs (aimiRNA) placed within the intron of ffgLUC...
2018: Plant Methods
Joseph Fennell, Charles Veys, Jose Dingle, Joachim Nwezeobi, Sharon van Brunschot, John Colvin, Bruce Grieve
Background: The paper introduces a multispectral imaging system and data-processing approach for the identification and discrimination of morphologically indistinguishable cryptic species of the destructive crop pest, the whitefly Bemisia tabaci . This investigation and the corresponding system design, was undertaken in two phases under controlled laboratory conditions. The first exploited a prototype benchtop variant of the proposed sensor system to analyse four cryptic species of whitefly reared under similar conditions...
2018: Plant Methods
Shuxian Li
Background: Phomopsis seed decay (PSD) of soybean ( Glycine max L. Merr.) is caused primarily by the seed-borne fungal pathogen Phomopsis longicolla T. W. Hobbs. The PSD disease reduces seed quality and yield worldwide. Development of effective techniques to evaluate soybean for resistance to PSD can facilitate identification of new sources of host resistance to manage the disease. This study was undertaken to develop and utilize a rapid cut-seedling inoculation technique to evaluate soybean genotypes for resistance to P...
2018: Plant Methods
William T Salter, Matthew E Gilbert, Thomas N Buckley
Background: Existing methods for directly measuring photosynthetic capacity ( A max ) have low throughput, which creates a key bottleneck for pre-breeding and ecological research. Currently available commercial leaf gas exchange systems are not designed to maximize throughput, on either a cost or time basis. Results: We present a novel multiplexed semi-portable gas exchange system, OCTOflux, that can measure A max with approximately 4-7 times the throughput of commercial devices, despite a lower capital cost...
2018: Plant Methods
Carmen Quiñonero López, Patricia Corral, Bénédicte Lorrain-Lorrette, Karen Martinez-Swatson, Franck Michoux, Henrik Toft Simonsen
Background: Thapsigargin and nortrilobolide are sesquiterpene lactones found in the Mediterranean plant Thapsia garganica L. Thapsigargin is a potent inhibitor of the sarco/endoplasmic reticulum calcium ATPase pump, inducing apoptosis in mammalian cells. This mechanism has been used to develop a thapsigargin-based cancer drug first by GenSpera and later Inspyr Therapeutics (Westlake Village, California). However, a stable production of thapsigargin is not established. Results: In vitro regeneration from leaf explants, shoot multiplication and rooting of T...
2018: Plant Methods
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