A mini binary vector series for plant transformation.

C Xiang; P Han; I Lutziger; K Wang; D J Oliver

doi:10.1023/a:1006201910593

Home/Archive/PubMed

PubMed · 10480394

A mini binary vector series for plant transformation.

C Xiang·P Han·I Lutziger·K Wang·D J Oliver

View original source ↗Export metadata ↓

Abstract

A streamlined mini binary vector was constructed that is less than 1/2 the size of the pBIN19 backbone (3.5 kb). This was accomplished by eliminating over 5 kb of non-T-DNA sequences from the pBIN19 vector. The vector still retains all the essential elements required for a binary vector. These include a RK2 replication origin, the nptIII gene conferring kanamycin resistance in bacteria, both the right and left T-DNA borders, and a multiple cloning site (MCS) in between the T-DNA borders to facilitate cloning. Due to the reduced size, more unique restriction sites are available in the MCS, thus allowing more versatile cloning. Since the traF region was not included, it is not possible to mobilize this binary vector into Agrobacterium by triparental mating. This problem can be easily resolved by direct transformation. The mini binary vector has been demonstrated to successfully transform Arabidopsis plants. Based on this mini binary vector, a series of binary vectors were constructed for plant transformation.

Explore related subjects

Agrobacterium tumefaciens Arabidopsis Blotting, Southern Cloning, Molecular DNA, Bacterial DNA, Plant Genetic Vectors Molecular Sequence Data Plants Sequence Analysis, DNA Transformation, Genetic Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

C Xiang, P Han, I Lutziger, K Wang, D J Oliver. 1999. A mini binary vector series for plant transformation.. https://doi.org/10.1023/a%3A1006201910593

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2026

The near-complete genome sequence of vB_AgrS_573, a lytic Agrobacterium tumefaciens phage isolated from karst cave sediment.

Phage vB_AgrS_573, isolated from oligotrophic karst cave sediment, is a lytic phage that infects Agrobacterium tumefaciens GV3101. It belongs to the class Caudoviricetes and harbors a 55,316-bp linear double-stranded DNA genome with 91 predicted protein-coding genes and a GC content of 52.19%; no transfer RNA genes were identified.

Agrobacterium tumefaciens↗

PubMed2007

Rhizobitoxine production in Agrobacterium tumefaciens C58 by Bradyrhizobium elkanii rtxACDEFG genes.

We examined the genetic basis and transfer for production of rhizobitoxine, an inhibitor of ethylene biosynthesis in plants, directed by the rtx genes of Bradyrhizobium elkanii. Comparison with genome sequences of Bradyrhizobium japonicum and Xanthomonas oryzae suggests that the rtx genes extend from the previously identified rtxAC genes through four additional genes rtxDEFG. Reverse transcription-PCR analysis showed that the rtxACDEFG genes are expressed as an operon. Mutational analysis indicated that rtxDEG mutants reduced rhizobitoxine biosynthesis, while the rtxA gene is essential for its synthesis. Introduction of the rtxACDEFG into Agrobacterium tumefaciens resulted in strong expression of rtxACDEFG and production of RtxA protein, but no rhizobitoxine was detectable. Addition of O-acetylhomoserine, a precursor of rhizobitoxine, to the Agrobacterium derivative, however, fostered production of rhizobitoxine in culture. The diluted culture supernatant inhibited the activities of beta-cystathionase and 1-aminocyclopropane-1-carboxylate synthase, indicating that A. tumefaciens carrying rtxACDEFG genes excreted biologically active rhizobitoxine.

Agrobacterium tumefaciens↗

PubMed2007

Generation of transgenic Lolium temulentum plants by Agrobacterium tumefaciens-mediated transformation.

Lolium temulentum L. (Darnel ryegrass) has been proposed to be used as a model species for functional genomics studies in forage and turf grasses, because it is a self-fertile, diploid species with a short life cycle and is closely related to other grasses. Embryogenic calluses were induced from mature embryos of a double haploid line developed through anther culture. The calluses were broken up into small pieces and used for Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harboring pCAMBIA1301 and pCAMBIA1305.2 vectors were used to infect embryogenic callus pieces. Hygromycin was used as a selection agent in stable transformation experiments. Hygromycin resistant calluses were obtained after 4-6 weeks of selection and transgenic plants were produced in 10-13 weeks after Agrobacterium-mediated transformation. Fertile plants were readily obtained after transferring the transgenics to the greenhouse. Transgenic nature of the regenerated plants was demonstrated by Polymerase chain reaction (PCR), Southern hybridization analysis, and GUS staining. Progeny analysis showed Mendelian inheritance of the transgenes. The transformation system provides a valuable tool for functionality tests of candidate genes in forage and turf grasses.

Agrobacterium tumefaciens↗