Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotiana tabacum

TransgenicNicotiana tabacumwith tolerance to 2,4-D has previously been produced using a bacterial 2,4-D-dioxygenase gene (tfdA) driven by the 35S promoter of cauliflower mosaic virus. Using promoters from thePisum sativumplastocyanin gene (petE) and anArabidopsis thalianahistone gene (H4A), we demonstrate that similar protection from 2,4-D can be obtained in transgenicN. tabacumby targeting expression oftfdAto either meristematic tissues or chloroplast-containing tissues. As with the 35S promoter constructs, the plants are tolerant but not completely resistant; very young seedlings in particular are only slightly protected. However, the levels of tolerance observed could offer a useful degree of protection from accidental spray drift.

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Unique Shine–Dalgarno Sequences in Cyanobacteria and Chloroplasts Reveal Evolutionary Differences in Their Translation Initiation

Genome Biology and Evolution ◽

10.1093/gbe/evz227 ◽

2019 ◽

Vol 11 (11) ◽

pp. 3194-3206 ◽

Cited By ~ 3

Author(s):

Yulong Wei ◽

Xuhua Xia

Keyword(s):

Arabidopsis Thaliana ◽

Nicotiana Tabacum ◽

Translation Initiation ◽

Ribosomal Proteins ◽

Selection Pressure ◽

Rna Seq ◽

Prominent Feature ◽

Transcriptomic Data ◽

Reduced Genome ◽

Cyanobacterial Genes

Abstract Microorganisms require efficient translation to grow and replicate rapidly, and translation is often rate-limited by initiation. A prominent feature that facilitates translation initiation in bacteria is the Shine–Dalgarno (SD) sequence. However, there is much debate over its conservation in Cyanobacteria and in chloroplasts which presumably originated from endosymbiosis of ancient Cyanobacteria. Elucidating the utilization of SD sequences in Cyanobacteria and in chloroplasts is therefore important to understand whether 1) SD role in Cyanobacterial translation has been reduced prior to chloroplast endosymbiosis or 2) translation in Cyanobacteria and in plastid has been subjected to different evolutionary pressures. To test these alternatives, we employed genomic, proteomic, and transcriptomic data to trace differences in SD usage among Synechocystis species, Microcystis aeruginosa, cyanophages, Nicotiana tabacum chloroplast, and Arabidopsis thaliana chloroplast. We corrected their mis-annotated 16S rRNA 3′ terminus using an RNA-Seq-based approach to determine their SD/anti-SD locational constraints using an improved measurement DtoStart. We found that cyanophages well-mimic Cyanobacteria in SD usage because both have been under the same selection pressure for SD-mediated initiation. Whereas chloroplasts lost this similarity because the need for SD-facilitated initiation has been reduced in plastids having much reduced genome size and different ribosomal proteins as a result of host-symbiont coevolution. Consequently, SD sequence significantly increases protein expression in Cyanobacteria but not in chloroplasts, and only Cyanobacterial genes compensate for a lack of SD sequence by having weaker secondary structures at the 5′ UTR. Our results suggest different evolutionary pressures operate on translation initiation in Cyanobacteria and in chloroplast.

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Construction of Two Vectors for a Bypass of Monocotyledon Plants Photorespiration

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.340.351 ◽

2011 ◽

Vol 340 ◽

pp. 351-356

Author(s):

Xue Liang Bai ◽

Dan Wang ◽

Ning Ning Liu ◽

Li Jing Wei ◽

Ye Rong Zhu ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Transgenic Plants ◽

Binary Vector ◽

Transit Peptide ◽

Expression Vectors ◽

Chloroplast Transit Peptide ◽

E Coli ◽

Plant Expression ◽

Glycolate Dehydrogenase ◽

Save Energy

In order to modify the photorespiration of monocotyledonous crops, we aimed to construct vectors that will be used to introduce a bypass to the native photorespiration pathway. Firstly, we cloned the encoding sequences of glyoxylate carboligase (GCL) and tartronic semialdehyde reductase (TSR) fromE. coli, glycolate dehydrogenase (GDH) fromArabidopsis thalianaand chloroplast transit peptide (cTP) from rice. Then we constructed a universal vector pEXP harboring the encoding sequence of cTP for targeting a protein into chloroplast. By insertion of these three encoding sequences into the universal vector pEXP, we obtained the expression cassettes for GCL, TSR and GDH, respectively. Finally, we inserted the cassettes for GCL and TSR in tandem into the binary vector pCAMBIA 1301, and for GDH into another binary vector, pPGN, to obtain our plant expression vectors pCAMBIA 1301-TG and pPGN-GDH, respectively. These two expression vectors possess different selection resistance and can be used to transform monocots together, to introduce the bypass pathway of photorespiration. By this way, the transgenic plants can recycle glycolate, the by-product of photosynthesis in C3plants, within the chloroplast, simultaneously, save energy and avoid the loss of ammonia, which will contribute to improved growth.

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Agroinfection of cloned Sri Lankan cassava mosaic virus DNA to Arabidopsis thaliana, Nicotiana tabacum and cassava

Archives of Virology ◽

10.1007/s00705-008-0238-z ◽

2008 ◽

Vol 153 (11) ◽

pp. 2149-2155 ◽

Cited By ~ 13

Author(s):

Dheeraj Mittal ◽

Basanta Kumar Borah ◽

Indranil Dasgupta

Keyword(s):

Arabidopsis Thaliana ◽

Nicotiana Tabacum ◽

Mosaic Virus ◽

Sri Lankan ◽

Cassava Mosaic Virus

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Growth modulation effects of CBM2a under the control of AtEXP4 and CaMV35S promoters in Arabidopsis thaliana, Nicotiana tabacum and Eucalyptus camaldulensis

Transgenic Research ◽

10.1007/s11248-017-0015-4 ◽

2017 ◽

Vol 26 (4) ◽

pp. 447-463 ◽

Cited By ~ 3

Author(s):

Pornthep Keadtidumrongkul ◽

Anongpat Suttangkakul ◽

Phitsanu Pinmanee ◽

Kanokwan Pattana ◽

Chokchai Kittiwongwattana ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Nicotiana Tabacum ◽

Eucalyptus Camaldulensis ◽

Growth Modulation

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A binary vector-based large insert library for Brassica napus and identification of clones linked to a fertility restorer locus for Ogura cytoplasmic male sterility (CMS)

Genome ◽

10.1139/g99-104 ◽

2000 ◽

Vol 43 (1) ◽

pp. 102-109 ◽

Cited By ~ 1

Author(s):

Yongzhong Wu ◽

Lomas Tulsieram ◽

Quangzhou Tao ◽

Hong-Bin Zhang ◽

Steven J Rothstein

Keyword(s):

Brassica Napus ◽

Cytoplasmic Male Sterility ◽

Male Sterility ◽

Plant Transformation ◽

Transformation Efficiency ◽

Binary Vector ◽

Brassica Species ◽

Chromosome Walking ◽

Fertility Restorer ◽

Plant Transformation Vector

We constructed and characterized a large DNA insert library for Brassica napus that would facilitate genome-related research and map-based cloning efforts in Brassica species. This library, consisting of 92 160 clones arrayed in 384-well microtiter dishes, was based on a conventional plant transformation vector (binary vector), and was constructed using a single ligation with transformation efficiency of over 5000 recombinants per microliter of ligation mixture. Every clone in this library contains an insert in the size range of 30-190 kb, facilitating both chromosome walking and plant transformation. Screening this library with three DNA markers (C2, F10, and CabR) that are linked to a fertility restorer locus for Ogura cytoplasmic male sterility (CMS) identified at least 17 positive clones for each probe. Among the 17 positive clones identified by C2, nine are linked to the restorer locus. Marker F10 identified 21 clones, of which only two are linked to the restorer locus. None of 68 clones identified by CabR is linked to the restorer locus. A stability test using two clones identified by the C2 marker indicated that large DNA inserts are stable in this conventional vector in both Escherichia coli and Agrobacterium. Key words: Brassica napus, binary vector, large DNA insert library, restorer gene.

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