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Andrew S Fister, Lena Landherr, Siela N Maximova, Mark J Guiltinan
Theobroma cacao , the source of cocoa, suffers significant losses to a variety of pathogens resulting in reduced incomes for millions of farmers in developing countries. Development of disease resistant cacao varieties is an essential strategy to combat this threat, but is limited by sources of genetic resistance and the slow generation time of this tropical tree crop. In this study, we present the first application of genome editing technology in cacao, using Agrobacterium-mediated transient transformation to introduce CRISPR/Cas9 components into cacao leaves and cotyledon cells...
2018: Frontiers in Plant Science
Guirong Tang, Qiong Li, Shenghui Xing, Ningning Li, Zheng Tang, Liangliang Yu, Junhui Yan, Xuan Li, Li Luo
Agrobacterium tumefaciens infects and causes crown galls in dicot plants by transferring T-DNA from the Ti plasmid to the host plant via a type IV secretion system (T4SS). This process requires appropriate environmental conditions, certain plant secretions and bacterial regulators. In our previous work, a member of the LysR family of transcriptional regulators (LsrB) in Sinorhizobium meliloti was found to modulate its symbiotic interactions with the host plant alfalfa. However, the function of its homologue in A...
March 16, 2018: Molecular Plant-microbe Interactions: MPMI
Jia Fang, Peng Nan, Zongying Gu, Xiaochun Ge, Yu-Qi Feng, Bao-Rong Lu
Transgenic glyphosate-tolerant plants overproducing EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) may exhibit enhanced fitness in glyphosate-free environments. If so, introgression of transgenes overexpressing EPSPS into wild relative species may lead to increased competitiveness of crop-wild hybrids, resulting in unpredicted environmental impact. Assessing fitness effects of transgenes overexpressing EPSPS in a model plant species can help address this question, while elucidating how overproducing EPSPS affects the fitness-related traits of plants...
2018: Frontiers in Plant Science
Longzheng Chen, Wei Li, Lorenzo Katin-Grazzini, Jing Ding, Xianbin Gu, Yanjun Li, Tingting Gu, Ren Wang, Xinchun Lin, Ziniu Deng, Richard J McAvoy, Frederick G Gmitter, Zhanao Deng, Yunde Zhao, Yi Li
Developing CRISPR/Cas9-mediated non-transgenic mutants in asexually propagated perennial crop plants is challenging but highly desirable. Here, we report a highly useful method using an Agrobacterium -mediated transient CRISPR/Cas9 gene expression system to create non-transgenic mutant plants without the need for sexual segregation. We have also developed a rapid, cost-effective, and high-throughput mutant screening protocol based on Illumina sequencing followed by high-resolution melting (HRM) analysis. Using tetraploid tobacco as a model species and the phytoene desaturase ( PDS ) gene as a target, we successfully created and expediently identified mutant plants, which were verified as tetra-allelic mutants...
2018: Horticulture Research
C R Gedling, E M Ali, A Gunadi, J J Finer, K Xie, Y Liu, N Yoshikawa, F Qu, A E Dorrance
Background: Virus induced gene silencing (VIGS) is a powerful genomics tool for interrogating the function of plant genes. Unfortunately, VIGS vectors often produce disease symptoms that interfere with the silencing phenotypes of target genes, or are frequently ineffective in certain plant genotypes or tissue types. This is especially true in crop plants like soybean [ Glycine max (L.) Merr]. To address these shortcomings, we modified the inoculation procedure of a VIGS vector based on Apple latent spherical virus (ALSV)...
2018: Plant Methods
Georg Hölzl, Christian Sohlenkamp, Miguel Angel Vences-Guzmán, Nicolas Gisch
Amino acid-containing acyloxyacyl lipids are composed of a 3-hydroxy fatty acid amide-bound to the α-amino group of an amino acid. A second fatty acid is ester-linked to the 3-hydroxy group of the first fatty acid. Most commonly, ornithine is the headgroup of these lipids, but glycine, serineglycine, glutamine and lysine have also been described in bacteria. Ornithine lipids (OL) can be synthesized by about 50% of the sequenced bacterial species, and several covalent modifications of its basic structure have been described...
March 7, 2018: Chemistry and Physics of Lipids
Samuel T Coradetti, Dominic Pinel, Gina Geiselman, Masakazu Ito, Stephen Mondo, Morgann C Reilly, Ya-Fang Cheng, Stefan Bauer, Igor Grigoriev, John M Gladden, Blake A Simmons, Rachel Brem, Adam P Arkin, Jeffrey M Skerker
The basidiomycete yeast Rhodosporidium toruloides (a.k.a. Rhodotorula toruloides ) accumulates high concentrations of lipids and carotenoids from diverse carbon sources. It has great potential as a model for the cellular biology of lipid droplets and for sustainable chemical production. We developed a method for high-throughput genetics (RB-TDNAseq), using sequence-barcoded Agrobacterium tumefaciens T-DNA insertions. We identified 1337 putative essential genes with low T-DNA insertion rates. We functionally profiled genes required for fatty acid catabolism and lipid accumulation, validating results with 35 targeted deletion strains...
March 9, 2018: ELife
Chenwen Xiao, Ling Li, Limin Lao, Yan Liu, Qiang Wei, Quan'an Ji, Guochang Sun, Fucheng Lin, Jiaoyu Wang, Guolian Bao
Trichophyton mentagrophytes is a fungus that causes skin disease in humans and other animals worldwide. Studies on molecular biology and fluorescent labeling of the fungus are limited. Here, we applied mCherry for the first time in T. mentagrophytes to label the fungus and its organelles. We constructed four expression vectors of mCherry or mCherry fusions containing a variety of resistance markers and promoters, which were then integrated, together with two previous mCherry expression vectors, in T. mentagrophytes via Agrobacterium tumefaciens-mediated transformation (AtMT)...
March 1, 2018: FEMS Microbiology Letters
Rongmei Li, Shuo Wang, Ronghua Sun, Xiang He, Yongting Liu, Congfeng Song
Transcription activator-like (TAL) effectors encoded by tal genes were recognized as a key virulence strategy used by Xanthomonas oryzae pv. oryzae (Xoo) to cause bacterial leaf blight of rice. TAL effector PthXo3 is a major virulence factor identified in a Philippine Xoo strain PXO61, and it can induce the expression of susceptibility gene OsSWEET14 by binding to the effector-binding element (EBE) in the promoter region. In this study, pthXo3 homologous genes were also identified and isolated from Xoo Chinese strain OS198 and Japanese strain JXOV, which were named as PthXo3OS198 and PthXo3JXOV respectively...
March 4, 2018: FEMS Microbiology Letters
Jesús Martínez-Cruz, Diego F Romero, Fernando de la Torre, Dolores Fernández-Ortuño, Juan Antonio Torés, Antonio de Vicente, Alejandro Perez-Garcia
Podosphaera xanthii is the main causal agent of powdery mildew disease in cucurbits. In a previous study, we determined that P. xanthii expresses approximately 50 Podosphaera effector candidates (PECs), identified based on the presence of a predicted signal peptide and the absence of functional annotation. In this work, we used host-induced gene silencing (HIGS), employing Agrobacterium tumefaciens as a vector for the delivery of the silencing constructs (ATM-HIGS), to identify genes involved in early plant-pathogen interaction...
March 7, 2018: Molecular Plant-microbe Interactions: MPMI
Hang Su, Yun-Tong Jiao, Fang-Fang Wang, Yue-E Liu, Wei-Li Niu, Guo-Tian Liu, Yan Xu
Putrescine and spermidine increase the transformation efficiency of Vitis vinifera L. cv. Thompson seedless. Accumulation of VpPR10.1 in transgenic V. vinifera Thompson seedless, likely increases its resistance to downy mildew. A more efficient method is described for facilitating Agrobacterium-mediated transformation of Vitis vinifera L. cv. Thompson Seedless somatic embryogenesis using polyamines (PAs). The efficacies of putrescine, spermidine and spermine are identified at a range of concentrations (10 µM, 100 µM and 1 mM) added to the culture medium during somatic embryo growth...
March 6, 2018: Plant Cell Reports
Abhishek Sharma, Priyanka Verma, Archana Mathur, Ajay Kumar Mathur
Terpenoid indole alkaloid (TIA) biosynthetic pathway of Catharanthus roseus possesses the major attention in current metabolic engineering efforts being the sole source of highly expensive antineoplastic molecules vinblastine and vincristine. The entire TIA pathway is fairly known at biochemical and genetic levels except the pathway steps leading to biosynthesis of catharanthine and tabersonine. To increase the in-planta yield of these antineoplastic metabolites for the pharmaceutical and drug industry, extensive plant tissue culture-based studies were performed to provide alternative production systems...
March 5, 2018: Protoplasma
Xiaotong Wang, Junkai Zhi, Xinru Liu, Hao Zhang, Huabo Liu, Jichen Xu
Heavy metal ATPase (HMA) plays an important role in phytoremediation via long-distance transportation from root to shoot. In this report, we identified a heavy metal ATPase gene, PtoHMA5, from Populus tomentosa Carr. Its encoded peptide consists of 967 amino acids and has eight trans-membrane motifs inside. Tobacco plants were transformed with this gene via Agrobacterium tumefaciens-mediated method. After exposure to 50 mg/LCdCl2 for 10 d, the transgenic lines displayed higher cadmium accumulation in leaves than did the wild-type plants with an absolute increase of 25...
February 28, 2018: International Journal of Biological Macromolecules
Han Ming Gan, Michael A Savka
This chapter presents a historical overview of the development and changes in scientific approaches to classifying members of the Agrobacterium genus. We also describe the changes in the inference of evolutionary relationships among Agrobacterium biovars and Agrobacterium strains from using the 16S rRNA marker to recA genes and to the use of multilocus sequence analysis (MLSA). Further, the impacts of the genomic era enabling low cost and rapid whole genome sequencing on Agrobacterium phylogeny are reviewed with a focus on the use of new and sophisticated bioinformatics approaches to refine phylogenetic inferences...
March 3, 2018: Current Topics in Microbiology and Immunology
Benoît Lacroix, Vitaly Citovsky
Besides the massive gene transfer from organelles to the nuclear genomes, which occurred during the early evolution of eukaryote lineages, the importance of horizontal gene transfer (HGT) in eukaryotes remains controversial. Yet, increasing amounts of genomic data reveal many cases of bacterium-to-eukaryote HGT that likely represent a significant force in adaptive evolution of eukaryotic species. However, DNA transfer involved in genetic transformation of plants by Agrobacterium species has traditionally been considered as the unique example of natural DNA transfer and integration into eukaryotic genomes...
March 3, 2018: Current Topics in Microbiology and Immunology
Tatiana V Matveeva
In most cases, the genetic engineering of plants uses Agrobacterium-mediated transformation to introduce novel genes. In nature, insertion of T-DNA into the plant genome and its subsequent transfer via sexual reproduction have been shown for several species in the genera Nicotiana, Ipomoea , and Linaria . A sequence homologous to T-DNA of the Ri plasmid of Agrobacterium rhizogenes was found in the genome of wild-type Nicotiana glauca (section Noctiflorae) more than 30 years ago and was named "cellular T-DNA" (cT-DNA)...
March 3, 2018: Current Topics in Microbiology and Immunology
Po-Yuan Shih, Shu-Jen Chou, Caroline Müller, Barbara Ann Halkier, Rosalia Deeken, Erh-Min Lai
Agrobacterium tumefaciens is the causal agent of crown gall disease in a wide range of plants via a unique interkingdom DNA transfer from bacterial cells into the plant genome. A. tumefaciens is capable of transferring its T-DNA into different plant parts at different developmental stages for transient and stable transformation. However, the plant genes and mechanisms involved in these transformation processes are not well understood. We used Arabidopsis thaliana Col-0 seedlings to reveal the gene expression profiles at early time points during Agrobacterium infection...
March 2, 2018: Molecular Plant Pathology
Fan-Chen Huang, Bi-Ju Fu, Yin-Tzu Liu, Yao-Ren Chang, Shin-Fei Chi, Pei-Ru Chien, Si-Chi Huang, Hau-Hsuan Hwang
Agrobacterium tumefaciens can genetically transform various eukaryotic cells because of the presence of a resident tumor-inducing (Ti) plasmid. During infection, a defined region of the Ti plasmid, transfer DNA (T-DNA), is transferred from bacteria into plant cells and causes plant cells to abnormally synthesize auxin and cytokinin, which results in crown gall disease. T-DNA and several virulence (Vir) proteins are secreted through a type IV secretion system (T4SS) composed of T-pilus and a transmembrane protein complex...
February 24, 2018: International Journal of Molecular Sciences
Kourosh Mohammadi, Ali Movahedi, Samaneh Sadat Maleki, Weibo Sun, Jiaxin Zhang, Amir Almasi Zadeh Yaghuti, Saeed Nourmohammadi, Qiang Zhuge
Drought and salinity are two main abiotic stressors that can disrupt plant growth and survival. Various biotechnological approaches have been used to alleviate the problem of drought stress by improving water stress resistance in forestry and agriculture. The drought sensitive 1 (DRS1) gene acts as a regulator of drought stress, identified in human, yeast and some model plants, such as Arabidopsis thaliana, but there have been no reports of DRS1 transformation in poplar plants to date. In this study, we transformed the DRS1 gene from Populus trichocarpa into Populus deltoides × Populus euramericana 'Nanlin895' using Agrobacterium tumefaciens-mediated transformation...
February 21, 2018: Plant Physiology and Biochemistry: PPB
Chao Fan, Xinxin Xu, Liya Song, Weishi Guan, Jinyang Li, Bo Liu, Pengjun Shi, Wei Zhang
A transfer DNA (T-DNA)-tagged mutant library of Humicola insolens was screened for mutants with altered cellulase production using the plate-clearing zone assay. Three selected mutants (5-A7, 5-C6, and 13-B7) exhibited significantly depressed FPase, CMCase and xylanase activities compared with the wild-type strain upon shake-flask fermentation, while the pNPCase and pNPGase activities of the three mutants were relatively higher than those of the parental strain. Combined with the results of SDS-PAGE and mass spectrometry, we suggest that expression of the CMCases Cel6B, Cel7B, CMC3, and XynA/B/C was reduced in the mutant strains...
March 2018: 3 Biotech
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