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United States Department of Agriculture

Agricultural Research Service

Title: Soybean Transformation Via the Pollen Tube Pathway

Authors
item Li, Zenglu - CROPSCI UOFI URBANA
item Nelson, Randall
item Widholm, Jack - CROPSCI UOFI URBANA
item Bent, Andrew - U WISCONSIN MAIDSON

Submitted to: Soybean Genetics Newsletter
Publication Type: Other
Publication Acceptance Date: December 20, 2002
Publication Date: December 20, 2003
Citation: LI, Z., NELSON, R.L., WIDHOLM, J.M., BENT, A. SOYBEAN TRANSFORMATION VIA THE POLLEN TUBE PATHWAY. SOYBEAN GENETICS NEWSLETTER. 2003. v. 29.

Technical Abstract: The pollen tube pathway method of transformation has been reported to be successful in several crops including soybeans. DNA can be transferred by cutting the stigma following pollination and applying the DNA solution on the severed style. 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 plasm ids pBI221 carrying a CaMV35S promoter-beta glucuronidase (gus) gene-nos terminator gene fusion construct and pBI0122 carrying a CaMV35S promoter-bar-g73' terminator gene fusion constructs were used. Approximately 5,000 seeds were produced from the flowers treated with DNA. None of the seeds from the plants treated with DNA containing the bar gene were found to be as resistant as the positive control. Morphological variation was observed in some plants, but this was not observed in the following generation. Approximately two percent of the seeds from plants treated with DNA containing the gus gene had a positive reaction in the GUS assay, however, no GUS activity was observed in the plant leaves from those seeds. Less than three percent of progeny seeds tested expressed a positive reaction and polymerase chain reaction (PCR) with seedling DNA did not detect the gus gene. Thus, we were unable to confirm the positive results of others when we used screen able marker genes.

Last Modified: 9/20/2014
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