Introduction of herbicide resistant gene (bar) into maize (Zea mays L.) via pollen tube pathway

来源 :Journal of Northeast Agricultural University(English Edition | 被引量 : 0次 | 上传用户:yoyo1028
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The pollen tube pathway method of transformation has been reported to be successful in most crops,but less successfu in maize.DNA can be transferred by cutting the stigma following pollination and applying the DNA solution in a suitable period DNA presumably reaches the ovary by flowing down the pollen tube and then integrates into the just fertilized but undivided zygotic cells.To provide the molecular evidence for this procedure,the plasmids pGBIRC carrying a CaMV35S promoter-PPT acetyle transferase(bar)gene-nos terminator gene fusion construct were used.Total 3 276 seeds were produced from the ears treated with DNA.It was found that 35 seedlings were GUS assay positive,but less intense than that of the positive controls,of which 17 were PCR amplification positive.But,only 13 of the seeds from the plants treated with DNA containing the bar gene were found to be resistant compared with the negative control.Less than 1.07% of progeny seedlings tested expressed a herbicide positive reaction and polymerase chain reaction(PCR)with seedling DNA did detect the bar gene.Morphological variation was observed in six plants.We succeed in obtain PPT-resistant maize inbred lines via pollen tube pathway. The pollen tube pathway method of transformation has been reported to be successful in most crops, but less successfull maize. DNA can be transferred by cutting the stigma following pollination and applying the DNA solution in a suitable period DNA presumably reaches the ovary by flowing down the pollen tube and then integrates into the just fertilized but undivided zygotic cells. To provide the molecular evidence for this procedure, the plasmids pGBIRC carrying a CaMV35S promoter-PPT acetyle transferase (bar) gene-nos terminator gene fusion construct were used. Total 3 276 seeds were produced from the ears treated with DNA.It was found that 35 seedlings were GUS assay positive, but less intense than that of the positive controls, of which 17 were PCR amplification positive. But, only 13 of the seeds from the plants treated with DNA containing the bar gene were found to be resistant compared with the negative control. Less than 1.07% of progeny seedlings demonstrated expressed a herbicide positive reac tion and polymerase chain reaction (PCR) with seedling DNA did detect the bar gene. Morphological variation was observed in six plants. We succeed in obtaining PPT-resistant maize inbred lines via pollen tube pathway.
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