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https://www.readbyqxmd.com/read/28928381/crispr-cas9-induced-targeted-mutagenesis-and-gene-replacement-to-generate-long-shelf-life-tomato-lines
#1
Qing-Hui Yu, Baike Wang, Ning Li, Yaping Tang, Shengbao Yang, Tao Yang, Juan Xu, Chunmiao Guo, Peng Yan, Qiang Wang, Patiguli Asmutola
Quickly and precisely gain genetically enhanced breeding elites with value-adding performance traits is desired by the crop breeders all the time. The present of gene editing technologies, especially the CRISPR/Cas9 system with the capacities of efficiency, versatility and multiplexing provides a reasonable expectation towards breeding goals. For exploiting possible application to accelerate the speed of process at breeding by CRISPR/Cas9 technology, in this study, the Agrobacterium tumefaciens-mediated CRISPR/Cas9 system transformation method was used for obtaining tomato ALC gene mutagenesis and replacement, in absence and presence of the homologous repair template...
September 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28923688/expression-and-activity-analysis-of-%C3%AE-gallinacin-3-in-arabidopsis
#2
Libo Jin, Yunpeng Wang, Nuo Xu, Dezhong Wang, Xiuming Liu, Renyi Peng, Chao Jiang, Xiaokun Li
β Gallinacin-3 (β Gal-3) is an antimicrobial peptide with strong antibacterial activity against Escherichia coli, Staphylococcus aureus and Salmonella typhimurium. In this study, the β Gal-3 gene was transferred into a plant genome by genetic engineering techniques. These transgenic plants can be used as feed additives to prevent poultry diseases and them might replace the antibiotics used in poultry industry. To ensure the β Gal-3 expresses effectively in Arabidopsis seeds, the expression was driven by promoter Ppha cloned from the β-phaseolin storage protein gene...
September 16, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28922840/quantification-of-agrobacterium-tumefaciens-c58-attachment-to-arabidopsis-thaliana-roots
#3
Anna Petrovicheva, Jessica Joyner, Theodore R Muth
Agrobacterium tumefaciens is the causal agent of crown gall disease and is a vector for DNA transfer in transgenic plants. The transformation process by A. tumefaciens has been widely studied, but the attachment stage has not been well characterized. Most measurements of attachment have used microscopy and colony counting, both of which are labor and time intensive. To reduce the time and effort required to analyze bacteria attaching to plant tissues, we developed a quantitative real-time PCR (qPCR) assay to quantify attached A...
October 2, 2017: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/28918530/exogenous-and-endogenous-increase-in-fungal-ggpp-increased-fungal-taxol-production
#4
Sameh S M Soliman, Kareem A Mosa, Ali A El-Keblawy, Mohamed I Husseiny
Taxol is an anticancer identified in both endophytic fungus and its host plant. Plant Taxol is a diterpenoid with geranylgeranyl diphosphate (GGPP) mediates the biosynthesis of its terpenoid moiety. Previous report has suggested that fungal Taxol may require terpenoid pathway for its biosynthesis. Here in this study, feeding a Taxol-producing endophytic fungus (Paraconiothyrium SSM001) with terpenoid precursors including isopentenyl pyrophosphate (IPP, isoprene) and GGPP enhanced Taxol production threefold and fivefold, respectively, compared to the control...
September 16, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28913803/doubled-haploid-transgenic-wheat-lines-by-microspore-transformation
#5
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/28913798/agrobacterium-mediated-transformation-of-wheat-using-immature-embryos
#6
Goetz Hensel, Cornelia Marthe, Jochen Kumlehn
Methods of the Agrobacterium-mediated transformation of bread wheat (Triticum aestivum L.) have been improved in recent years so that genetic engineering can be routinely used for functional genomics as well as for wheat breeding. In the protocol described here, immature embryos of the spring-type model genotype Bobwhite SH 98 26 have been used. Preculture and temperature pretreatment of these explants have led to the reproducible generation of transgenic plants at efficiencies between 5 and 15%. Whereas primary transgenic plants regenerated in vitro commonly show reduced fitness and fertility, no apparent variations with regard to morphology and grain set in their transgenic progeny as compared to wild-type counterparts were observed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28912104/dissection-of-membrane-binding-and-remodeling-regions-in-two-classes-of-bacterial-phospholipid-n-methyltransferases
#7
Linna Danne, Meriyem Aktas, Nadine Grund, Tim Bentler, Ralf Erdmann, Franz Narberhaus
Bacterial phospholipid N-methyltransferases (Pmts) catalyze the formation of phosphatidylcholine (PC) via successive N-methylation of phosphatidylethanolamine (PE). They are classified into Sinorhizobium-type and Rhodobacter-type enzymes. The Sinorhizobium-type PmtA protein from the plant pathogen Agrobacterium tumefaciens is recruited to anionic lipids in the cytoplasmic membrane via two amphipathic helices called αA and αF. Besides its enzymatic activity, PmtA is able to remodel membranes mediated by the αA domain...
September 11, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28911019/early-arabidopsis-root-hair-growth-stimulation-by-pathogenic-strains-of-pseudomonas-syringae
#8
Tamara Pecenková, Martin Janda, Jitka Ortmannová, Vladimíra Hajná, Zuzana Stehlíková, Viktor Žárský
Background and Aims: Selected beneficial Pseudomonas spp. strains have the ability to influence root architecture in Arabidopsis thaliana by inhibiting primary root elongation and promoting lateral root and root hair formation. A crucial role for auxin in this long-term (1week), long-distance plant-microbe interaction has been demonstrated. Methods: Arabidopsis seedlings were cultivated in vitro on vertical plates and inoculated with pathogenic strains Pseudomonas syringae pv...
September 1, 2017: Annals of Botany
https://www.readbyqxmd.com/read/28906473/engineered-disease-resistance-in-cotton-using-rna-interference-to-knock-down-cotton-leaf-curl-kokhran-virus-burewala-and-cotton-leaf-curl-multan-betasatellite-expression
#9
Aftab Ahmad, Muhammad Zia-Ur-Rehman, Usman Hameed, Abdul Qayyum Rao, Ammara Ahad, Aneela Yasmeen, Faheem Akram, Kamran Shahzad Bajwa, Jodi Scheffler, Idrees Ahmad Nasir, Ahmad Ali Shahid, Muhammad Javed Iqbal, Tayyab Husnain, Muhammad Saleem Haider, Judith K Brown
Cotton leaf curl virus disease (CLCuD) is caused by a suite of whitefly-transmitted begomovirus species and strains, resulting in extensive losses annually in India and Pakistan. RNA-interference (RNAi) is a proven technology used for knockdown of gene expression in higher organisms and viruses. In this study, a small interfering RNA (siRNA) construct was designed to target the AC1 gene of Cotton leaf curl Kokhran virus-Burewala (CLCuKoV-Bu) and the βC1 gene and satellite conserved region of the Cotton leaf curl Multan betasatellite (CLCuMB)...
September 14, 2017: Viruses
https://www.readbyqxmd.com/read/28905515/crispr-cas9-mediated-efficient-targeted-mutagenesis-in-grape-in-the-first-generation
#10
Xianhang Wang, Mingxing Tu, Dejun Wang, Jianwei Liu, Yajuan Li, Zhi Li, Yuejin Wang, Xiping Wang
The clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system is a powerful tool for editing plant genomes. Efficient genome editing of grape (Vitis vinifera) suspension cells using the type II CRISPR/Cas9 system has been demonstrated; however, it has not been established whether this system can be applied to get biallelic mutations in the first generation of grape. In this current study, we designed four guide RNAs for the VvWRKY52 transcription factor gene for using with the CRISPR/Cas9 system, and obtained transgenic plants via Agrobacterium-mediated transformation, using somatic embryos of the Thompson Seedless cultivar...
September 14, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28904411/structural-and-functional-properties-biosynthesis-and-patenting-trends-of-bacterial-succinoglycan-a-review
#11
REVIEW
Urmi Halder, Aparna Banerjee, Rajib Bandopadhyay
The exopolysaccharide succinoglycan is produced mainly by a large number of soil microbes of Agrobacterium, Rhizobium or Pseudomonas genera etc. Structural properties of succinoglycan are unique in terms of its thermal stability and superior viscosifying property. Unlike the other highly commercialized bacterial exopolysaccharides like dextran or xanthan, mass scale application of succinoglycan has not been that much broadly explored yet. Bacterial succinoglycan is found suitable as a viscosifying and emulsifying agent in food industry, in gravel packing or fluid-loss control agent etc...
September 2017: Indian Journal of Microbiology
https://www.readbyqxmd.com/read/28901382/construction-of-a-chalcone-reductase-expression-vector-and-transformation-of-soybean-plants
#12
Nan Wu, Pi-Wu Wang, Nan Lin, Shi Lu, Yong-Qi Feng, Jie Rong, Zhuo Zhang, Jing Qu
The present study aimed to clone the soybean chalcone reductase 3 (CHR3) and create a recombinant expression vector pCAMBIA3300‑CHR3 containing Bar resistance gene as a selection marker, and then obtain transgenic soybean plants using Agrobacterium infection. The plant expression vector pCAMBIA3300‑CHR3 was transferred into soybean receptor plants, Jinong 17 and Jilin 30. Polymerase chain reaction (PCR) and Southern blotting were used to confirm the positive transgenic plants. Additionally, reverse transcription‑quantitative PCR (RT‑qPCR) was used to detect CHR3 expression and isoliquiritigenin content was measured using high‑performance liquid chromatography (HPLC) in the transgenic offspring...
August 29, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28899486/immobilization-on-graphene-oxide-improves-the-thermal-stability-and-bioconversion-efficiency-of-d-psicose-3-epimerase-for-rare-sugar-production
#13
Samir R Dedania, Manisha J Patel, Dijit M Patel, Rekha C Akhani, Darshan H Patel
D-Psicose (D-ribo-2-hexulose or D-allulose), an epimer of D-fructose is considered as a rare low-calorie sugar displaying important physiological functions. Enzymatic production using ketose 3-epimerases is the feasible process for the production of D-Psicose. However, major drawbacks in application of ketose 3-epimerases are bioconversion efficiency and reusability of the enzyme. We have attempted immobilization of ketose 3-epimerases from Agrobacterium tumefaciens (agtu) D-psicose 3-epimerase (DPEase) on graphene oxide...
December 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28895654/graft-accelerated-virus-induced-gene-silencing-facilitates-functional-genomics-in-rose-flowers
#14
Huijun Yan, Shaochuan Shi, Nan Ma, Xiaoqian Cao, Hao Zhang, Xianqin Qiu, Qigang Wang, Hongying Jian, Ningning Zhou, Zhao Zhang, Kaixue Tang
Rose has emerged as a model ornamental plant for studies of flower development, senescence, and morphology, as well as the metabolism of floral fragrances and colors. Virus-induced gene silencing (VIGS) has long since been used in functional genomics studies of rose by vacuum infiltration of cuttings or seedlings with an Agrobacterium suspension carrying TRV-derived vectors. However, VIGS in rose flowers remains a challenge because of its low efficiency and long time to silencing. Here we present a novel and rapid VIGS method that can be used to analyze gene function in rose, called 'graft-accelerated VIGS', where axillary sprouts are cut off the rose plant and vacuum infiltrated with Agrobacterium...
September 12, 2017: Journal of Integrative Plant Biology
https://www.readbyqxmd.com/read/28893289/recombinant-expression-of-beak-and-feather-disease-virus-capsid-protein-and-assembly-of-virus-like-particles-in-nicotiana-benthamiana
#15
Guy L Regnard, Edward P Rybicki, Inga I Hitzeroth
BACKGROUND: Beak and feather disease virus (BFDV) is an important disease causing agent affecting psittacines. BFDV is highly infectious and can present as acute, chronic or subclinical disease. The virus causes immunodeficiency and is often associated with secondary infections. No commercial vaccine is available and yields of recombinant BFDV capsid protein (CP) expressed in insect cells and bacteria are yet to be seen as commercially viable, although both systems produced BFDV CP that could successfully assemble into virus-like particles (VLPs)...
September 11, 2017: Virology Journal
https://www.readbyqxmd.com/read/28878794/a-tau-class-glutathione-s-transferase-is-involved-in-trans-resveratrol-transport-out-of-grapevine-cells
#16
Ascensión Martínez-Márquez, María J Martínez-Esteso, María T Vilella-Antón, Susana Sellés-Marchart, Jaime A Morante-Carriel, Elias Hurtado, Javier Palazon, Roque Bru-Martínez
Vitis vinifera cell cultures respond to pathogens and elicitors by synthesizing and extracellularly accumulating stilbenoid phytoalexins. Large amounts of trans-resveratrol (t-R) are produced when a cell culture is elicited with methylated cyclodextrins (MBCD), either alone or combined with methyl jasmonate (MeJA). t-R transport to the extracellular medium, which represents the apoplastic space, would place this antifungal defense right in the battlefield to efficiently fight against pathogen attack. Yet despite their physiological relevance, these transport pathways are mostly unknown...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28878506/analysis-of-tandem-repeat-units-of-the-promoter-of-capsanthin-capsorubin-synthase-ccs-gene-in-pepper-fruit
#17
Shi-Lin Tian, Zheng Li, Li Li, S N M Shah, Zhen-Hui Gong
Capsanthin/capsorubin synthase (Ccs) gene is a key gene that regulates the synthesis of capsanthin and the development of red coloration in pepper fruits. There are three tandem repeat units in the promoter region of Ccs, but the potential effects of the number of repetitive units on the transcriptional regulation of Ccs has been unclear. In the present study, expression vectors carrying different numbers of repeat units of the Ccs promoter were constructed, and the transient expression of the β-glucuronidase (GUS) gene was used to detect differences in expression levels associated with the promoter fragments...
July 2017: Physiology and Molecular Biology of Plants: An International Journal of Functional Plant Biology
https://www.readbyqxmd.com/read/28878341/a-botrytis-cinerea-klp-7-kinesin-acts-as-a-virulence-determinant-during-plant-infection
#18
Pamil Tayal, Sumit Raj, Esha Sharma, Manoj Kumar, Vikram Dayaman, Nidhi Verma, Abhimanyu Jogawat, Meenakshi Dua, Rupam Kapoor, Atul Kumar Johri
Botrytis cinerea is a necrotrophic pathogen that infects many important crops. In an attempt to unravel some novel factors that govern pathogenicity in B. cinerea, Agrobacterium tumefaciens mediated transformation (ATMT) was deployed, and a number of tagged transformants were generated. Among these, a mutant, BCM-29 exhibited slower growth rate, reduced conidia size, conidiation and penetration. The mutant was also defective in secretion of oxalic acid (OA) and exhibited reduced activities of polygalacturonase (PG) and pectin methyl esterases (PME)...
September 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28874565/virulence-protein-vird5-of-agrobacterium-tumefaciens-binds-to-kinetochores-in-host-cells-via-an-interaction-with-spt4
#19
Xiaorong Zhang, G Paul H van Heusden, Paul J J Hooykaas
The bacterium Agrobacterium tumefaciens causes crown gall tumor formation in plants. During infection the bacteria translocate an oncogenic piece of DNA (transferred DNA, T-DNA) into plant cells at the infection site. A number of virulence proteins are cotransported into host cells concomitantly with the T-DNA to effectuate transformation. Using yeast as a model host, we find that one of these proteins, VirD5, localizes to the centromeres/kinetochores in the nucleus of the host cells by its interaction with the conserved protein Spt4...
September 5, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28874475/broad-host-range-expression-reveals-native-and-host-regulatory-elements-that-influence-heterologous-antibiotic-production-in-gram-negative-bacteria
#20
Jia Jia Zhang, Xiaoyu Tang, Michelle Zhang, Darlene Nguyen, Bradley S Moore
Heterologous expression has become a powerful tool for studying microbial biosynthetic gene clusters (BGCs). Here, we extend the transformation-associated recombination cloning and heterologous expression platform for microbial BGCs to include Gram-negative proteobacterial expression hosts. Using a broad-host-range expression platform, we test the implicit assumption that biosynthetic pathways are more successfully expressed in more closely related heterologous hosts. Cloning and expression of the violacein BGC from Pseudoalteromonas luteoviolacea 2ta16 revealed robust production in two proteobacterial hosts, Pseudomonas putida KT2440 and Agrobacterium tumefaciens LBA4404, but very little production of the antibiotic in various laboratory strains of Escherichia coli, despite their closer phylogenetic relationship...
September 5, 2017: MBio
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