barriers to sexual hybridization between cultivated and wild relatives, rescue of pollinated embryos has been used: when a cross yields a viable embryo but the surrounding seed endosperm4 is not viable, the embryo is taken from the nonviable seed environment and “rescued” by being grown in tissue culture.

Other techniques to introduce variation in cultivars include cell fusion, somaclonal variation, chemical or x-ray mutagenesis, and genetic engineering (see section 2.4.2). Cell fusion is used to produce novel combinations of genomic material from nuclei and organelles when plants are not sexually compatible (Ehlenfeldt and Helgeson 1987); it can be performed only on plants that can be cultured with protoplast technologies. With protoplast technologies, cells are disconnected from tissues, their walls are removed, and their membranes are prepared for fusion. Somaclonal variation is variation that occurs during the tissue-culture process, and its phenotypic outcomes are often similar to other forms of mutagenesis. Genetic engineering is the transfer of a or a few genes into a cultivar with the use of Agrobacterium tumefaciens, microprojectile bombardment, electroporation, or microinjection. Transgenic methods will be discussed in more detail in subsequent sections of this report.

One of the main differences among the techniques used for introducing variation is in the amount of DNA involved. In progeny resulting from a cross between two cultivars, half the genome comes from each cultivar. Each half (haploid) genome contains a significant amount of DNA (table 1.1). The amount transferred with conventional breeding in the case of Arabidopsis could be 70 megabases (Mb) (half the progeny's haploid genome comes from each parent). For bread wheat, the amount of DNA could be almost 8000 Mb. In contrast, transgenic methods involve the addition of only a few genes and flanking regulatory sequences (totaling about 1-20 kilobases).

Another important difference among the techniques can be the source of the transferred DNA. Sexual hybridization involves genes from sexually compatible species, which tend to be rather similar. Mutagenesis and the somaclonal variation process do not add genes, but rather modify existing genes. Cell fusion can add genes from evolutionarily divergent plant species (such as, plants from different genera), but normally fused cells are from somewhat related plants (for example, the technique has not been conducted by fusing cells from plants and microorganisms). In genetic engineering or transgenic methods, genes from any organism in the biosphere can be used as long as the regulatory sequences are functional in the host plant. For example with genetic engineering researchers

4  

Nutritional tissue in seeds.



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