共查询到20条相似文献，搜索用时 109 毫秒
1.
Lilian E Pino Simone Lombardi-Crestana Mariana S Azevedo Danielle C Scotton Lucélia Borgo Vera Quecini Antonio Figueira Lázaro EP Peres 《Plant methods》2010,6(1):23
Background
The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps. 相似文献2.
3.
Carvalho RF Campos ML Pino LE Crestana SL Zsögön A Lima JE Benedito VA Peres LE 《Plant methods》2011,7(1):18-14
Background
The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways.Results
We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system.Conclusions
The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk. 相似文献4.
A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts 总被引：2，自引：0，他引：2
Rebecca Bart Mawsheng Chern Chang-Jin Park Laura Bartley Pamela C Ronald 《Plant methods》2006,2(1):13-9
Background
Transient assays using protoplasts are ideal for processing large quantities of genetic data coming out of hi-throughput assays. Previously, protoplasts have routinely been prepared from dicot tissue or cell suspension cultures and yet a good system for rice protoplast isolation and manipulation is lacking. 相似文献5.
Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues 总被引：2，自引：0，他引：2
Anna Lytovchenko Romina Beleggia Nicolas Schauer Tal Isaacson Jan E Leuendorf Hanjo Hellmann Jocelyn KC Rose Alisdair R Fernie 《Plant methods》2009,5(1):4-11
Background
The concept of metabolite profiling has been around for decades and technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. However, studies are generally confined to polar compounds alone. Here we describe a simple method for lipophilic compounds analysis in various plant tissues. 相似文献6.
Kelvin T. Chiong Mona B. Damaj Carmen S. Padilla Carlos A. Avila Shankar R. Pant Kranthi K. Mandadi Ninfa R. Ramos Denise V. Carvalho T. Erik Mirkov 《Plant methods》2017,13(1):106
Background
Several high-throughput molecular genetic analyses rely on high-quality genomic DNA. Copurification of other molecules can negatively impact the functionality of plant DNA preparations employed in these procedures. Isolating DNA from agronomically important crops, such as sugarcane, rice, citrus, potato and tomato is a challenge due to the presence of high fiber, polysaccharides, or secondary metabolites. We present a simplified, rapid and reproducible SDS-based method that provides high-quality and -quantity of DNA from small amounts of leaf tissue, as required by the emerging biotechnology and molecular genetic applications.Results
We developed the TENS-CO method as a simplified SDS-based isolation procedure with sequential steps of purification to remove polysaccharides and polyphenols using 2-mercaptoethanol and potassium acetate, chloroform partitioning, and sodium acetate/ethanol precipitation to yield high-quantity and -quality DNA consistently from small amounts of tissue (0.15 g) for different plant species. The method is simplified and rapid in terms of requiring minimal manipulation, smaller extraction volume, reduced homogenization time (20 s) and DNA precipitation (one precipitation for 1 h). The method has been demonstrated to accelerate screening of large amounts of plant tissues from species that are rich in polysaccharides and secondary metabolites for Southern blot analysis of reporter gene overexpressing lines, pathogen detection by quantitative PCR, and genotyping of disease-resistant plants using marker-assisted selection.Conclusion
To facilitate molecular genetic studies in major agronomical crops, we have developed the TENS-CO method as a simple, rapid, reproducible and scalable protocol enabling efficient and robust isolation of high-quality and -quantity DNA from small amounts of tissue from sugarcane, rice, citrus, potato, and tomato, thereby reducing significantly the time and resources used for DNA isolation.7.
Background
Deep sequencing is a powerful tool for novel small RNA discovery. Illumina small RNA sequencing library preparation requires a pre-adenylated 3?? end adapter containing a 5??,5??-adenyl pyrophosphoryl moiety. In the absence of ATP, this adapter can be ligated to the 3?? hydroxyl group of small RNA, while RNA self-ligation and concatenation are repressed. Pre-adenylated adapters are one of the most essential and costly components required for library preparation, and few are commercially available.Results
We demonstrate that DNA oligo with 5?? phosphate and 3?? amine groups can be enzymatically adenylated by T4 RNA ligase 1 to generate customized pre-adenylated adapters. We have constructed and sequenced a small RNA library for tomato (Solanum lycopersicum) using the T4 RNA ligase 1 adenylated adapter.Conclusion
We provide an efficient and low-cost method for small RNA sequencing library preparation, which takes two days to complete and costs around $20 per library. This protocol has been tested in several plant species for small RNA sequencing including sweet potato, pepper, watermelon, and cowpea, and could be readily applied to any RNA samples. 相似文献8.
Virus-induced gene silencing as a tool for functional analyses in the emerging model plant Aquilegia (columbine, Ranunculaceae) 总被引：1，自引：0，他引：1
Background
There is considerable interest in rapid assays or screening systems for assigning gene function. However, analysis of gene function in the flowers of some species is restricted due to the difficulty of producing stably transformed transgenic plants. As a result, experimental approaches based on transient gene expression assays are frequently used. Biolistics has long been used for transient over-expression of genes of interest, but has not been exploited for gene silencing studies. Agrobacterium-infiltration has also been used, but the focus primarily has been on the transient transformation of leaf tissue. 相似文献9.
Naohiro Kato Dexter Reynolds Matthew L Brown Marietta Boisdore Yukichi Fujikawa Andrea Morales Lee A Meisel 《Plant methods》2008,4(1):9
Background
The isolation of green fluorescent protein (GFP) and the development of spectral variants over the past decade have begun to reveal the dynamic nature of protein trafficking and organelle motility. In planta analyses of this dynamic process have typically been limited to only two organelles or proteins at a time in only a few cell types. 相似文献10.
Background
The amount of DNA in the chloroplasts of some plant species has been shown recently to decline dramatically during leaf development. A high-throughput method of DNA detection in chloroplasts is now needed in order to facilitate the further investigation of this process using large numbers of tissue samples. 相似文献11.
Background
X-ray micro-CT has increasingly been used for 3D imaging of plant structures. At the micrometer resolution however, limitations in X-ray contrast often lead to datasets with poor qualitative and quantitative measures, especially within dense cell clusters of plant tissue specimens. The current study developed protocols for delivering a cesium based contrast enhancing solution to varying plant tissue specimens for the purpose of improving 3D tissue structure characterization within plant specimens, accompanied by new image processing workflows to extract the additional data generated by the contrast enhanced scans.Results
Following passive delivery of a 10% cesium iodide contrast solution, significant increases of 85.4 and 38.0% in analyzable cell volumes were observed in pear fruit hypanthium and tomato fruit outer mesocarp samples. A significant increase of 139.6% in the number of analyzable cells was observed in the pear fruit samples along the added ability to locate and isolate better brachysclereids and vasculature in the sample volume. Furthermore, contrast enhancement resulted in significant improvement in the definition of collenchyma and parenchyma in the petiolule of tomato leaflets, from which both qualitative and quantitative data can be extracted with respect to cell measures. However, contrast enhancement was not achieved in leaf vasculature and mesophyll tissue due to fundamental limitations. Active contrast delivery to apple fruit hypanthium samples did yield a small but insignificant increase in analyzable volume and cells, but data on vasculature can now be extracted better in correspondence to the pear hypanthium samples. Contrast delivery thus improved visualization and analysis the most in dense tissue types.Conclusions
The cesium based contrast enhancing protocols and workflows can be utilized to obtain detailed 3D data on the internal microstructure of plant samples, and can be adapted to additional samples of interest with minimal effort. The resulting datasets can therefore be utilized for more accurate downstream studies that requires 3D data.12.
Background
Recent developments, including the sequencing of a number of plant genomes, have greatly increased the amount of data available to scientists and has enabled high throughput investigations where many genes are investigated simultaneously. To perform these studies, recombinational cloning methods such as the Gateway system have been adapted to plant transformation vectors to facilitate the creation of overexpression, tagging and silencing vectors on a large scale. 相似文献13.
Berthold Heinze 《Plant methods》2007,3(1):4-7
Background
Chloroplast genomes evolve slowly and many primers for PCR amplification and analysis of chloroplast sequences can be used across a wide array of genera. In some cases 'universal' primers have been designed for the purpose of working across species boundaries. However, the essential information on these primer sequences is scattered throughout the literature. 相似文献14.
15.
Shufu Que Kuan Li Min Chen Yongfei Wang Qiaobin Yang Wenfeng Zhang Baoqian Zhang Bangshu Xiong Huaqin He 《Plant methods》2012,8(1):5
Background
As a result of the growing body of protein phosphorylation sites data, the number of phosphoprotein databases is constantly increasing, and dozens of tools are available for predicting protein phosphorylation sites to achieve fast automatic results. However, none of the existing tools has been developed to predict protein phosphorylation sites in rice. 相似文献16.
Background
Plant genome sequencing has resulted in the identification of a large number of uncharacterized genes. To investigate these unknown gene functions, several transient transformation systems have been developed as quick and convenient alternatives to the lengthy transgenic assay. These transient assays include biolistic bombardment, protoplast transfection and Agrobacterium-mediated transient transformation, each having advantages and disadvantages depending on the research purposes. 相似文献17.
Milene Miranda Praa Carlos Roberto Carvalho Wellington Ronildo Clarindo 《Scientia Horticulturae》2009,122(3):501-505
A relatively rapid, practical and reliable procedure for in vitro production of tetraploid tomato using colchicine is described. Using this procedure 11.11% generation of tetraploids was obtained after exposure of seedling shoot meristem explants with 8 mM colchicine for 96 h. Confirmation of tetraploid production (2n = 48), was determined by flow cytometry and verified by cytogenetic analysis of root tip preparations. The results indicate that the procedure is adequate for induction and screening of tetraploid tomato plantlets. 相似文献
18.
Kiyoon Kang Kyungjin Lee Sung-Oh Sohn Sangkyu Park Sungbeom Lee Soo Young Kim Young Soon Kim Kyoungwhan Back 《Scientia Horticulturae》2009,120(4):504-510
Tomato contains high levels of amines such as serotonin and tyramine and is a suitable host to enhance phenylpropanoid amides (PAs), an important class of nutraceuticals with strong antioxidant activity and chemotherapeutic effects, by ectopic expression of the corresponding gene, serotonin N-hydroxycinnamoyltransferase (SHT). To assess whether ectopic overexpression of SHT cDNA under the control of the CaMV 35S promoter would enhance levels of PAs, we generated transgenic tomato plants and analyzed the levels of PAs. Transgenic tomato plants exhibited increased synthesis of PAs such as feruloylserotonin (FS), 4-coumaroylserotonin (CS), feruloyltyramine (FT), 4-coumaroyltyramine (CT), and feruloyloctopamine (FO) in 1-month-old leaves compared to the wild type. The increase and relative levels of PAs were even more apparent in 3-month-old leaves of transgenic tomato. When tomato leaves were challenged by wounding, levels of PAs in the best transgenic line increased by 3- and 10-fold for CS + FS and CT + FT, respectively. In contrast to leaves, tomato fruit only showed enhanced synthesis of CT + FT, whereas CS + FS levels were not enhanced. Regarding amine content, the levels of tyramine were much higher than those of serotonin in tomato leaves and fruits. The high levels of tyramine may contribute to the preferential production of CT + FT rather than CS + FS, although SHT enzyme shows the highest substrate affinity toward serotonin rather than tyramine. 相似文献
19.
Background
High throughput DNA isolation from plants is a major bottleneck for most studies requiring large sample sizes. A variety of protocols have been developed for DNA isolation from plants. However, many species, including conifers, have high contents of secondary metabolites that interfere with the extraction process or the subsequent analysis steps. Here, we describe a procedure for high-throughput DNA isolation from conifers. 相似文献20.
Qin Li Zhongwen Huang Meng Xu Chenguang Wang Junyi Gai Youjun Huang Xiaoming Pang Rongling Wu 《Plant methods》2010,6(1):13