Engineered selective plant male sterility through pollen-specific expression of the EcoRI restriction endonuclease.
Unintended gene flow from transgenic plants via pollen, seed and vegetative propagation is a regulatory concern because of potential admixture in food and crop systems, as well as hybridization and introgression to wild and weedy relatives. Bioconfinement of transgenic pollen would help address some of these concerns and enable transgenic plant production for several crops where gene flow is an issue. Here, we demonstrate the expression of the restriction endonuclease EcoRI under the control of the tomato pollen-specific LAT52 promoter is an effective method for generating selective male sterility in Nicotiana tabacum (tobacco). Of nine transgenic events recovered, four events had very high bioconfinement with tightly controlled EcoRI expression in pollen and negligible-to-no expression other plant tissues. Transgenic plants had normal morphology wherein vegetative growth and reproductivity were similar to nontransgenic controls. In glasshouse experiments, transgenic lines were hand-crossed to both male-sterile and emasculated nontransgenic tobacco varieties. Progeny analysis of 16 000-40 000 seeds per transgenic line demonstrated five lines approached (>99.7%) or attained 100% bioconfinement for one or more generations. Bioconfinement was again demonstrated at or near 100% under field conditions where four transgenic lines were grown in close proximity to male-sterile tobacco, and 900-2100 seeds per male-sterile line were analysed for transgenes. Based upon these results, we conclude EcoRI-driven selective male sterility holds practical potential as a safe and reliable transgene bioconfinement strategy. Given the mechanism of male sterility, this method could be applicable to any plant species.
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Reginald J
RJ
Millwood
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Hong S
HS
Moon
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Charleson R
CR
Poovaiah
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Balasubramaniam
B
Muthukumar
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
John Hollis
JH
Rice
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Jason M
JM
Abercrombie
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Laura L
LL
Abercrombie
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
William Derek
WD
Green
Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
Charles Neal
CN
Stewart
D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D051456:Gene Flow; D005818:Genetic Engineering; D006824:Hybridization, Genetic; D009928:Organ Specificity; D051479:Plant Infertility / Q000235:genetics*; D030821:Plants, Genetically Modified; D011058:Pollen / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012639:Seeds / Q000235:genetics; D014026:Nicotiana / Q000235:genetics*; D019076:Transgenes
D015246:Deoxyribonuclease EcoRI
Eco
RI; bioconfinement; gene flow; pollen; selective male sterility; transgene