Techniques in plant molecular biology – progress and problems

Abstract

Progress in plant molecular biology has been dependent on efficient methods of introducing foreign DNA into plant cells. Gene transfer into plant cells can be achieved by either direct uptake of DNA or the natural process of gene transfer carried out by the soil bacterium Agrobacterium. Versatile gene‐transfer vectors have been developed for use with Agrobacterium and more recently vectors based on the genomes of plant viruses have become available. Using this technology the expression of foreign DNA, the functional analysis of plant DNA sequences, the investigation of the mechanism of viral DNA replication and cell to cell spread, as well as the study of transposition, can be carried out. In addition, the versatility of the gene‐transfer vectors is such that they may be used to isolate genes not amenable to isolation using conventional protocols. This review concentrates on these aspects of plant molecular biology and discusses the limitations of the experimental systems that are currently available.

Transformation of cauliflower (Brassica oleracea L. var. botrytis) ? an experimental survey

“…Often regenerated plants revealed an altered morphology. This might be due to insertional mutagenesis of gene coding for morphological traits or position effects [29]. Insertional mutagenesis could also be the reason for failure in regenerating transformed cauliflower calli in our experiments, although this has not been tested yet.…”

Systemic expression of a bacterial gene by a tobacco mosaic virus-based vector.

“…Hybrid RNAs containing both TMV-U1 and the inserted bacterial gene sequences were encapsidated by the odontoglossum ringspot virus coat protein, facilitating their transmission and amplification on pgin to subsequent plants. The vector TB2 provides a rapid means of expressing genes and gene variants in plants.The expression of foreign genes in plants has proven advantageous to the study of molecular biology (1,2). Stable gene transfer to whole plants can be achieved by a combination of DNA transformation and tissue culture techniques or by virus transfection.…”

“…The expression of foreign genes in plants has proven advantageous to the study of molecular biology (1,2). Stable gene transfer to whole plants can be achieved by a combination of DNA transformation and tissue culture techniques or by virus transfection.…”

“…Stable gene transfer to whole plants can be achieved by a combination of DNA transformation and tissue culture techniques or by virus transfection. With viral-based vectors the ease of infection avoids the time-consuming procedures, "position" effects, and "somaclonal" variation seen when foreign genes are integrated into the plant genome (1). The merits of using a systemically expressing plant RNA virus-based vector have been extensively reviewed (3)(4)(5)(6)(7).…”

Mechanical vs Agroinfiltration: Efficient mechanism of plant RNA virus infection

“…Even a robust approach like agrobacterium mediated gene transfer doesn't come without limitations, as it is limited to dicotyledonous plants and is not amenable for monocots [26]. Moreover, there is difference in the virulence of strains for Agrobacterium tumefaciens, and requires extensive optimization of technique as a protocol for transformation differs from one species to another [25,27].…”