pSiM24 Is a Novel Versatile Gene Expression Vector for Transient Assays As Well As Stable Expression of Foreign Genes in Plants

Sahoo, Dipak Kumar; Dey, Nrisingha; Maiti, Indu B.

doi:10.1371/journal.pone.0098988

Cited by 15 publications

(9 citation statements)

References 83 publications

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“…These include constitutive viral, bacterial (CmYLCV, M24, FMV, and nos), or plant (AtUbi10, OsAct1, PvUbi1, and PvUbi2) promoters, which provide a range of expression strengths in a variety of plant species (Stavolone et al, 2003;Sahoo et al, 2014;Maiti et al, 1997;Norris et al, 1993;McElroy et al, 1990;Mann et al, 2012). We have also included the Arabidopsis Ec1.2 and YAO promoters, which are egg cell and embryo sac/pollen specific, respectively.…”

Section: Discussionmentioning

confidence: 99%

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

et al. 2017

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We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination. Regulation of transcription is also possible. A Web-based tool streamlines vector selection and construction. One advantage of our platform is the use of the Csy-type (CRISPR system yersinia) ribonuclease 4 (Csy4) and tRNA processing enzymes to simultaneously express multiple guide RNAs (gRNAs). For example, we demonstrate targeted deletions in up to six genes by expressing 12 gRNAs from a single transcript. Csy4 and tRNA expression systems are almost twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promoters. Mutagenesis can be further enhanced 2.5-fold by incorporating the Trex2 exonuclease. Finally, we demonstrate that Cas9 nickases induce gene targeting at frequencies comparable to native Cas9 when they are delivered on geminivirus replicons. The reagents have been successfully validated in tomato (Solanum lycopersicum), tobacco (Nicotiana tabacum), Medicago truncatula, wheat (Triticum aestivum), and barley (Hordeum vulgare).

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Section: Discussionmentioning

confidence: 99%

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

et al. 2017

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“…Approximately 149.9 g of MDIPU, the demethylated metabolite of IPU by PdmAB, was detected in the medium planted with (32), and a terminator of the tobacco nopaline synthase gene (tNos). The pdmB expression cassette contains the enhancer of the figwort mosaic virus 35S gene (eFMV) (48), the promoter of the Brassica CBP1 gene (prBrCBP) (reference patent, CN201310724357), the chloroplast transit peptide-coding sequence (AtCTP) (32), and a terminator of the pea rbcSE9 gene (tPsE9) (49). The phosphinothricin (glufosinate) N-acetyltransferase gene (pat) expression cassette for the selection of transgenic lines contains the promoter of the cauliflower mosaic virus 35S gene (pr35s), the pat gene from Streptomyces viridochromogenes, and the terminator of the cauliflower mosaic virus 35S gene (t35S).…”

Section: Resultsmentioning

confidence: 99%

Enhanced and Complete Removal of Phenylurea Herbicides by Combinational Transgenic Plant-Microbe Remediation

Yan

Huang

et al. 2018

Appl Environ Microbiol

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The synergistic relationships between plants and their rhizospheric microbes can be used to develop a combinational bioremediation method, overcoming the constraints of individual phytoremediation or bioaugmentation method. Here, we provide a combinational transgenic plant-microbial remediation system for a more efficient removal of phenylurea herbicides (PHs) from contaminated-sites. The transgenic synthesizing the bacterial-demethylase PdmAB in the chloroplast was developed. The constructed transgenic exhibited significant tolerance to isoproturon (IPU), a typical PH, and it took up the IPU through roots and transported to leaves, where the majority of the IPU was demethylated to 3-(4-isopropylphenyl)-1-methylurea (MDIPU). The produced intermediate was released outside of roots and further metabolized by the combinationally inoculated MDIPU-mineralizing bacterium sp. strain 1017-1 in the rhizosphere, resulting in an enhanced and complete removal of IPU from soil. Mutual benefits were built for both transgenic and strain 1017-1. The transgenic offered strain 1017-1 a suitable accommodation and in return, the latter protected the plant from the phytotoxicity of MDIPU. The biomass of the transgenic and the residence of the inoculated degrading microbes in the combinational treatment increased significantly compared to that in their respective individual transgenic plant treatment or bioaugmentation treatment. The influence of the structure of bacterial community by combinational treatment was between that of the two individual treatments. Overall, the combination of two approaches, phytoremediation by transgenic plants and bioaugmentation with intermediate-mineralizing microbes in the rhizosphere, represents an innovative strategy for the enhanced and complete remediation of pollutant-contaminated sites. Phytoremediation of organic pollutant-contaminated sites using transgenic plants expressing bacterial enzyme has been well described. The major constraint of transgenic plants transferred with a single catabolic gene is that they can also accumulate/release intermediates, still causing phytotoxicity or additional environmental problems. On the other hand, bioaugmentation with degrading strains also has its drawbacks including instability of the inoculated strains and low bioavailability of pollutants. In this study, the synergistic relationship between transgenic expressing the bacterial-demethylase PdmAB in the chloroplast and the inoculated intermediate-mineralizing bacterium sp. strain 1017-1 in the rhizosphere is used to develop an intriguing bioremediation method. The combinational transgenic plant-microbe remediation system shows a more efficient and complete removal of phenylurea herbicides from contaminated-sites, and can overcome the constraints of individual phytoremediation or bioaugmentation methods.

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“…S‐6‐6, from S. Gowda, UF, FL); (12) Nicotiana benthamiana (‘Labstrain’ from E. Hiatt, LSBC‐KY); (13) Nicotiana tabacum cv KY14; (14) Nicotiana tabacum cv Samsun NN; (15) Nicotiana tabacum cv Petit Havana; (16) Nicotiana tabacum cv Xanthi; (17) Nicotiana tabacum cv Xanthi xcnn; (18) Nicotiana tabacum cv Hybrid 49‐50 (19) Nicotiana tabacum cv Hybrid 110; (20) Nicotiana tabacum cv Hybrid 111; (21) Nicotiana tabacum cv Hybrid 112. Plants were grown in a greenhouse under standard condition as described in previous publication …”

Section: Methodsmentioning

confidence: 99%