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Claudia D Norzagaray-Valenzuela, Lourdes J Germán-Báez, Marco A Valdez-Flores, Sergio Hernández-Verdugo, Luke M Shelton, Angel Valdez-Ortiz
Microalgae are photosynthetic microorganisms widely used for the production of highly valued compounds, and recently they have been shown to be promising as a system for the heterologous expression of proteins. Several transformation methods have been successfully developed, from which the Agrobacterium tumefaciens-mediated method remains the most promising. However, microalgae transformation efficiency by A. tumefaciens is shown to vary depending on several transformation conditions. The present study aimed to establish an efficient genetic transformation system in the green microalgae Dunaliella tertiolecta using the A...
May 16, 2018: Journal of Microbiological Methods
Léon Otten
The transfer of T-DNA sequences from Agrobacterium to plant cells is a well-understood process of natural genetic engineering. The expression of T-DNA genes in plants leads to tumors, hairy roots, or transgenic plants. The transformed cells multiply and synthesize small molecules, called opines, used by Agrobacteria for their growth. Several T-DNA genes stimulate or influence plant growth. Among these, iaaH and iaaM encode proteins involved in auxin synthesis, whereas ipt encodes a protein involved in cytokinin synthesis...
May 17, 2018: Current Topics in Microbiology and Immunology
Paul J J Hooykaas, G Paul H van Heusden, Xiaolei Niu, M Reza Roushan, Jalal Soltani, Xiaorong Zhang, Bert J van der Zaal
Two decades ago, it was discovered that the well-known plant vector Agrobacterium tumefaciens can also transform yeasts and fungi when these microorganisms are co-cultivated on a solid substrate in the presence of a phenolic inducer such as acetosyringone. It is important that the medium has a low pH (5-6) and that the temperature is kept at room temperature (20-25 °C) during co-cultivation. Nowadays, Agrobacterium-mediated transformation (AMT) is the method of choice for the transformation of many fungal species; as the method is simple, the transformation efficiencies are much higher than with other methods, and AMT leads to single-copy integration much more frequently than do other methods...
May 17, 2018: Current Topics in Microbiology and Immunology
Guillaume P Robin, Jochen Kleemann, Ulla Neumann, Lisa Cabre, Jean-Félix Dallery, Nicolas Lapalu, Richard J O'Connell
The genome of the hemibiotrophic anthracnose fungus, Colletotrichum higginsianum , encodes a large inventory of putative secreted effector proteins that are sequentially expressed at different stages of plant infection, namely appressorium-mediated penetration, biotrophy and necrotrophy. However, the destinations to which these proteins are addressed inside plant cells are unknown. In the present study, we selected 61 putative effector genes that are highly induced in appressoria and/or biotrophic hyphae. We then used Agrobacterium -mediated transformation to transiently express them as N -terminal fusions with fluorescent proteins in cells of Nicotiana benthamiana for imaging by confocal microscopy...
2018: Frontiers in Plant Science
Tania Ho-Plágaro, Raúl Huertas, María I Tamayo-Navarrete, Juan A Ocampo, José M García-Garrido
Background: Solanum lycopersicum , an economically important crop grown worldwide, has been used as a model for the study of arbuscular mycorrhizal (AM) symbiosis in non-legume plants for several years and several cDNA array hybridization studies have revealed specific transcriptomic profiles of mycorrhizal tomato roots. However, a method to easily screen candidate genes which could play an important role during tomato mycorrhization is required. Results: We have developed an optimized procedure for composite tomato plant obtaining achieved through Agrobacterium rhizogenes -mediated transformation...
2018: Plant Methods
Shraddha Shukla, Kashyap Kumar Dubey
Coenzyme Q10 (CoQ) or ubiquinone is found in the biological system which is synthesized by the conjugation of benzoquinone ring with isoprenoid chain of variable length. Coenzyme Q10 supplementation energizes the body and increases body energy production in the form of ATP and helps to treat various human diseases such as cardiomyopathy, muscular dystrophy, periodontal disease, etc. Reports of these potential therapeutic advantages of CoQ10 have resulted in its high market demand, which focus the researchers to work on this molecule and develop better bioprocess methods for commercial level production...
May 2018: 3 Biotech
Xindi Li, Yanning Wang, Sha Chen, Huiqin Tian, Daqi Fu, Benzhong Zhu, Yunbo Luo, Hongliang Zhu
Numerous studies have been focusing on breeding tomato plants with enhanced lycopene accumulation, considering its positive effects of fruits on the visual and functional properties. In this study, we used a bidirectional strategy: promoting the biosynthesis of lycopene, while inhibiting the conversion from lycopene to β- and α-carotene. The accumulation of lycopene was promoted by knocking down some genes associated with the carotenoid metabolic pathway. Finally, five genes were selected to be edited in genome by CRISPR/Cas9 system using Agrobacterium tumefaciens -mediated transformation...
2018: Frontiers in Plant Science
Hye Jeong Kim, Hyun Suk Cho, Jung Hun Pak, Tackmin Kwon, Jai-Heon Lee, Doh-Hoon Kim, Dong Hee Lee, Chang-Gi Kim, Young-Soo Chung
Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from T2 seedlings...
May 10, 2018: Molecules and Cells
Hai-Lei Wei, Wei Zhang, Alan Collmer
The bacterial pathogen Pseudomonas syringae pv. tomato DC3000 suppresses the two-tiered innate immune system of Nicotiana benthamiana and other plants by injecting a complex repertoire of type III secretion effector (T3E) proteins. Effectorless polymutant DC3000D36E was used with a modularized system for native delivery of the 29 DC3000 T3Es singly and in pairs. Assays of the performance of this T3E library in N. benthamiana leaves revealed a matrix of T3E interplay, with six T3Es eliciting death and eight others variously suppressing the death activity of the six...
May 8, 2018: Cell Reports
Yunfu Gu, Yingyan Wang, Yihao Sun, Ke Zhao, Quanju Xiang, Xiumei Yu, Xiaoping Zhang, Qiang Chen
BACKGROUND: Alleviating arsenic (As) contamination is a high-priority environmental issue. Hyperaccumulator plants may harbor endophytic bacteria able to detoxify As. Therefore, we investigated the distribution, diversity, As (III) resistance levels, and resistance-related functional genes of arsenite-resistant bacterial endophytes in Pteris vittata L. growing in a lead-zinc mining area with different As contamination levels. RESULTS: A total of 116 arsenite-resistant bacteria were isolated from roots of P...
May 8, 2018: BMC Microbiology
Bin Zhang, Bingyi Guo, Yunlong Bai, Huizhe Lu, Yanhong Dong
Novel azamacrolides comprising the triazole moiety were synthesized and evaluated for their quorum sensing inhibitor activities on the Agrobacterium tumefaciens . It was found that the inhibition rate of compound Z12 - 3 at 200 mg/L (0.45 mM) can reach 67%. The potential binding modes between these molecules and the TraR QS receptor was performed by molecular docking. The results showed that the two nitrogen atoms in the triazole ring of Z12 - 3 formed hydrogen bonds with GLN-2, and the carbonyl group (C=O) in the amide formed hydrogen bonds with water...
May 4, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Xueyu Zhang, Shaokui Zheng, Hangyu Zhang, Shoupeng Duan
This study clarified the dominant nitrogen (N)-transformation pathway and the key ammonia-oxidizing microbial species at three loading levels during optimization of the anoxic/oxic (A/O) process for sewage treatment. Comprehensive N-transformation activity analysis showed that ammonia oxidization was performed predominantly by aerobic chemolithotrophic and heterotrophic ammonia oxidization, whereas N2 production was performed primarily by anoxic denitrification in the anoxic unit. The abundances of ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria, and anaerobic AOB in activated sludge reflected their activities on the basis of high-throughput sequencing data...
April 30, 2018: Bioresource Technology
Jiaying Sun, Ruidi Xu, Shuqin Xiao, Yuanyuan Lu, Qifeng Zhang, Chunsheng Xue
Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used in filamentous fungi. In this study, an efficient Agrobacterium tumefaciens-mediated transformation approach was developed for the plant pathogenic fungus, Kabatiella zeae, the causative pathogen of eyespot in maize. Five parameters were selected to optimize efficiencies of transformation. A. tumefaciens concentration, conidia concentration of K. zeae and mixing ratio of A. tumefaciens and K. zeae were found to exert a significant influence on all parameters...
May 3, 2018: Journal of Microbiological Methods
Qian Du, Xiangdong Yang, Jinhua Zhang, Xiaofang Zhong, Kyung Seok Kim, Jing Yang, Guojie Xing, Xiaoyu Li, Zhaoyuan Jiang, Qiyun Li, Yingshan Dong, Hongyu Pan
Phytophthora root and stem rot (PRR) caused by Phytophthora sojae is one of the most devastating diseases reducing soybean (Glycine max) production all over the world. Harpin proteins in many plant pathogenic bacteria were confirmed to enhance disease and insect resistance in crop plants. Here, a harpin protein-encoding gene hrpZpsta from the P. syringae pv. tabaci strain Psta218 was codon-optimized (renamed hrpZm) and introduced into soybean cultivars Williams 82 and Shennong 9 by Agrobacterium-mediated transformation...
May 4, 2018: Transgenic Research
Tian Tian Cui, Yu Bin, Jian Hong Yan, Peng Ying Mei, Zhong An Li, Chang Yong Zhou, Zhen Song
Yellow vein clearing disease (YVCD) causes significant economic losses in lemon and other species of citrus. Usually, citrus yellow vein clearing virus (CYVCV) is considered to be the causal agent of YVCD. However, mixed infection of CYVCV and Indian citrus ringspot virus (ICRSV) or other pathogens is often detected in citrus plants with YVCD. In this study, we re-examined the causal agent of YVCD to fulfill Koch's postulates. First, the full-length genome of CYVCV isolate AY (CYVCV-AY) was amplified by long-distance RT-PCR from a Eureka lemon [Citrus limon (L) Brum...
May 4, 2018: Phytopathology
Guomin Han, Qian Shao, Cuiping Li, Kai Zhao, Li Jiang, Jun Fan, Haiyang Jiang, Fang Tao
Aspergillus flavus often invade many important corps and produce harmful aflatoxins both in preharvest and during storage stages. The regulation mechanism of aflatoxin biosynthesis in this fungus has not been well explored mainly due to the lack of an efficient transformation method for constructing a genome-wide gene mutant library. This challenge was resolved in this study, where a reliable and efficient Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for A. flavus NRRL 3357 was established...
May 2018: Journal of Microbiology / the Microbiological Society of Korea
Gulamnabi L Vanti, Ishwarappa S Katageri, Shashikala R Inamdar, Vamadevaiah Hiremathada, Bale M Swamy
Cotton is an important crop that is continuously cultivated around the world. However, its production has decreased in recent times due to wide ranging insects and also current practices of using synthetic insecticides that are not precise and their residues impairing the biodiversity. Hence, the search for newer classes of efficient entomotoxic proteins continues. Genetically modified cotton crops with cry genes from Bacillus thuringiensis, have been cultivated across the world, which overcome the chewing type insect menace...
April 28, 2018: Journal of Biotechnology
Yunxiao Liu, Xia Lan, Shiren Song, Ling Yin, Ian B Dry, Junjie Qu, Jiang Xiang, Jiang Lu
Downy mildew is one of the most destructive diseases of grapevine, causing tremendous economic loss in the grape and wine industry. The disease agent Plasmopara viticola is an obligate biotrophic oomycete, from which over 100 candidate RXLR effectors have been identified. In this study, 83 candidate RXLR effector genes ( PvRXLRs ) were cloned from the P. viticola isolate "JL-7-2" genome. The results of the yeast signal sequence trap assay indicated that most of the candidate effectors are secretory proteins...
2018: Frontiers in Plant Science
J Jansing, J F Buyel
The yield of recombinant proteins in plants determines their economic competitiveness as a production platform compared to microbes and mammalian cells. The promoter, untranslated regions (UTRs) and codon usage can all contribute to the yield, but potential interactions among these components have not been examined in detail. Here we investigated the effect of two promoters (35SS and nos) and four 5'UTRs on the spatiotemporal expression of DsRed mRNA and the accumulation of DsRed protein during transient expression in tobacco (Nicotiana tabacum) mediated by Agrobacterium tumefaciens...
April 27, 2018: Biotechnology Journal
Almudena González-Mula, Julien Lang, Catherine Grandclément, Delphine Naquin, Mohammed Ahmar, Laurent Soulère, Yves Queneau, Yves Dessaux, Denis Faure
Agrobacterium tumefaciens constructs an ecological niche in its host plant by transferring the T-DNA from its Ti plasmid into the host genome and by diverting the host metabolism. We combined transcriptomics and genetics for understanding the A. tumefaciens lifestyle when it colonizes Arabidopsis thaliana tumors. Transcriptomics highlighted: a transition from a motile to sessile behavior that mobilizes some master regulators (Hfq, CtrA, DivK and PleD); a remodeling of some cell surface components (O-antigen, succinoglucan, curdlan, att genes, putative fasciclin) and functions associated with plant defense (Ef-Tu and flagellin pathogen-associated molecular pattern-response and glycerol-3-phosphate and nitric oxide signaling); and an exploitation of a wide variety of host resources, including opines, amino acids, sugars, organic acids, phosphate, phosphorylated compounds, and iron...
April 27, 2018: New Phytologist
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