Genetic modification of the waxy character in barley under the action of exogenous DNA of the wild variety |
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| Authors: | NV Turbin VN Soyfer NA Kartel NM Chekalin YL Dorohov YB Titov KK Cieminis |
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| Institution: | 1. All-Union Institute of Applied Molecular Biology and Genetics of All-Union Academy of Agricultural Sciences, Moscow, g-117, USSR;2. Institute of Genetics and Cytology, Academy of Sciences of Byelorussian SSR, Minsk and Poltavian Agricultural Institute, Poltava, U.S.S.R. |
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| Abstract: | Genetic transformation was studied on spring barley mutants carrying the recessive mutant allele of waxy locus.Analysis of the pollen grains of 38 control plants that were not subjected to any treatment showed that in the whole sample there was no mutant grain that stained black with iodine. It is also indicative of the genotypical purity of the original waxy plants.After mechanical damage (puncture in the top and in the side of a grain) there were 5 pollen grains fo wild type among 124000 mutant grains. Injection of grains with 2 μl distilled water led to the emergence of pollen grains that stained black at a frequency similar to that after puncture. The overall frequency of wild barley pollen grains for all control variants was 2.2·10−5.The frequencies of wild-type pollen grains were practically the same after injections of DNA from E. coli, extensively deproteinized highly polymeric DNA from barley leaves or slightly deproteinized low polymeric DNA from barley endospermal material.There was no marked increase of the wild pollen frequency after the injection of slightly deproteinized low polymeric DNA from the endospernal material (2.2·10−5 in the control versus 3.7·10−5 in the experiment).The analysis of the material for the amount of altered pollen grains in invidual plants also unequivocally demonstrated significant differences between the control and the experiment. In the first four variants there was no plant having over these altered pollen grains among the 2500 mutant grains examined. In the variants with injections of barley DNA differing in the extent of deproteinization and in polymerisation only in one case (of low polymeric slightly deproteinized DNA) there were no plants with many altered pollen grains. In all the other variants there were plants having much pollen of wild type.The largest number of plants with a great many affected pollen grains occurred in the variant with highly polymeric DNA from the endospermal material not subjected to deproteinization with chloroform and isoamyl alcohol. |
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| Keywords: | SSC |
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