Award Abstract #9975715

Identification of Plant Genes Involved in Agrobacterium-Mediated Transformation

NSF Org: IOS Division of Integrative Organismal Systems

divider line
Initial Amendment Date: September 13, 1999
divider line
Latest Amendment Date: September 16, 2004
divider line
Award Number: 9975715
divider line
Award Instrument: Continuing grant
divider line
Program Manager: Jane Silverthorne
IOS Division of Integrative Organismal Systems
BIO Directorate for Biological Sciences
divider line
Start Date: September 1, 1999
divider line
Expires: August 31, 2005 (Estimated)
divider line
Awarded Amount to Date: $4,465,751.00
divider line
Investigator(s): Stanton Gelvin gelvin@bilbo.bio.purdue.edu (Principal Investigator)
Vitaly Citovsky (Co-Principal Investigator)

divider line
Sponsor: Purdue Research Foundation

West Lafayette, IN 47907- (317)494-6200
divider line
NSF Program(s): PLANT GENOME RESEARCH PROJECT
divider line
Program Reference Code(s): 9178, 9251, BIOT, 1228, 9179, 9109
divider line
Program Element Code(s): 1329

ABSTRACT

The objectives of this project are to identify plant genes involved in Agrobacterium-mediated genetic transformation, and subsequently to utilize this information to improve the genetic transformation of maize and to prevent crown gall disease on grape.

Agrobacterium-mediated transformation is the most popular method currently in use for generating transgenic plants. However, there are many aspects of the transformation process that are poorly understood. Specifically, little is known about the roles that host genes and proteins play in Agrobacterium-plant cell interactions. Understanding of the plant contribution to the transformation process will certainly result in new technologies for genetic manipulation of agronomically important species that are presently recalcitrant to Agrobacterium-mediated transformation. For example, many plants can be relatively efficiently transformed transiently by Agrobacterium, yet their stable transformation remains elusive. It is likely that these crops are deficient in T-DNA integration into the plant genome. An understanding of the plant genes involved in the integration process may allow scientists to devise strategies to over-express homologues of these genes in the recalcitrant species and alter them to transformation-competence. On the other hand, inactivation of the same genes in susceptible plants will make them crown gall-resistant. Crown gall disease affects numerous target crop species, and its prevention would be important for many sectors of the agricultural community.

This project involves an international collaboration of three laboratories (two in the US and one in Switzerland) and will use four approaches to identify and characterize plant genes involved in Agrobacterium-mediated transformation: 1) Screening of existing collections of T-DNA-tagged Arabidopsis mutants for rat (resistant to Agrobacterium transformation) phenotypes. These mutants will be investigated genetically and biochemically to identify the specific stage at which the transformation process is arrested; 2) Utilization of the yeast one- and two-hybrid systems to identify plant proteins that interact with Vir (virulence) proteins transferred to the plant by Agrobacterium. These Vir proteins include VirF, VirB2, VirD2, and VirE2; 3) Utilization of differential display, microarray, and subtraction approaches to identify plant genes whose expression is up- or down-regulated during early stages of Agrobacterium-mediated transformation; 4) Characterization of Arabidopsis genes involved in Agrobacterium-mediated transformation using reverse genetics, and examination of the functions of proteins encoded by these genes using specific biological assays developed in each of the collaborating laboratories.

The information gained from our analysis of Arabidopsis genes will be used to genetically modify maize such that it will become more susceptible to Agrobacterium-mediated transformation, and to genetically modify grape such that it will become resistant to crown gall disease.

Deliverables: E-mail the PI

Additional project information can be found at: http://www.bio.purdue.edu/courses/gelvinweb/gelvin.html

BOOKS/ONE TIME PROCEEDING

Crouzet, P., and Hohn, B.. "Transgenic Plants",&nbsp05/01/2000-05/01/2001, "Encyclopedia of Life Sciences", &nbsp2001, "MacMillan
www.els.net".

Ziemienowicz, A., Gorlich, D., rossi, L., Merkle, T., Tinland, B., and Hohn. B.. "Nuclear import and integration of Agrobacterium T-DNA in vitro.",&nbsp05/01/2000-05/01/2001, , P. de Wit, T. Bisseling, W. Stiekma"Proceedings of the MPMI Congress on the Biology of Plant-Microbe Interactions", &nbsp2000, "Vol. 2
International Society of Molecular Plant--Microbe Interactions, St. Paul, Minn USA
pp. 164-169.".

Crouzet, P., and Hohn, B.. "Transgenic Plants",&nbsp05/01/2001-05/01/2002, "Encyclopedia of Life Sciences", &nbsp2001, "MacMillan
www.els.net".

Ziemienowicz, A., Gorlich, D., rossi, L., Merkle, T., Tinland, B., and Hohn. B.. "Nuclear import and integration of Agrobacterium T-DNA in vitro.",&nbsp05/01/2001-05/01/2002, , P. de Wit, T. Bisseling, W. Stiekma"Proceedings of the MPMI Congress on the Biology of Plant-Microbe Interactions", &nbsp2000, "Vol. 2
International Society of Molecular Plant--Microbe Interactions, St. Paul, Minn USA
pp. 164-169.".

Crouzet, P., and Hohn, B.. "Transgenic Plants",&nbsp05/01/2002-05/01/2003, "Encyclopedia of Life Sciences", &nbsp2001, "MacMillan
www.els.net".

Ziemienowicz, A., Gorlich, D., rossi, L., Merkle, T., Tinland, B., and Hohn. B.. "Nuclear import and integration of Agrobacterium T-DNA in vitro.",&nbsp05/01/2002-05/01/2003, , P. de Wit, T. Bisseling, W. Stiekma"Proceedings of the MPMI Congress on the Biology of Plant-Microbe Interactions", &nbsp2000, "Vol. 2
International Society of Molecular Plant--Microbe Interactions, St. Paul, Minn USA
pp. 164-169.".

Wu, Y.-Q., and Hohn, B.. "Transfer to plants and integration into chromosomal DNA of T-DNA of Agrobacterium tumefaciens",&nbsp05/01/2002-05/01/2003, , G. Stacey and N.T. Keen"Plant-Microbe Interactions", &nbsp2003, "pp. 1-18".

Crouzet, P., and Hohn, B.. "Transgenic Plants",&nbsp09/01/1999-08/31/2005, "Encyclopedia of Life Sciences", &nbsp2001, "MacMillan
www.els.net".

Ziemienowicz, A., Gorlich, D., rossi, L., Merkle, T., Tinland, B., and Hohn. B.. "Nuclear import and integration of Agrobacterium T-DNA in vitro.",&nbsp09/01/1999-08/31/2005, , P. de Wit, T. Bisseling, W. Stiekma"Proceedings of the MPMI Congress on the Biology of Plant-Microbe Interactions", &nbsp2000, "Vol. 2
International Society of Molecular Plant--Microbe Interactions, St. Paul, Minn USA
pp. 164-169.".

Wu, Y.-Q., and Hohn, B.. "Transfer to plants and integration into chromosomal DNA of T-DNA of Agrobacterium tumefaciens",&nbsp09/01/1999-08/31/2005, , G. Stacey and N.T. Keen"Plant-Microbe Interactions", &nbsp2003, "pp. 1-18".

Please report errors in award information by writing to: awardsearch@nsf.gov.

Back to Top of page

FUNDING   AWARDS   DISCOVERIES   NEWS   PUBLICATIONS   STATISTICS   ABOUT NSF   FASTLANE  

Research.gov  |  USA.gov  |  National Science Board  |  Recovery Act  |  Budget and Performance  |  Annual Financial Report

Web Policies and Important Links  |  Privacy  |  FOIA  |  NO FEAR Act  |  Inspector General  |  Webmaster Contact  |  Site Map

National Science Foundation Logo

The National Science Foundation, 4201 Wilson Boulevard, Arlington, Virginia 22230, USA Tel: (703) 292-5111, FIRS: (800) 877-8339 | TDD: (800) 281-8749

Text Only Version

Select a optioninsert the text to search 

Top of page

---

Usablenet Assistive is a UsableNet product. Usablenet Assistive Main Page.