Presentation Title

Developing a Functional Genomics Tool for Cowpea

Faculty Mentor

Isgouhi Kaloshian

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 10:30 AM

Location

CREVELING 118

Session

POSTER 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

The cowpea plant (Vigna unguiculata) is a major source of nutrition in drought-ridden areas such as Sub-Saharan Africa. Like many crops, cowpea is plagued by numerous pests, one of which is the cowpea aphid (Aphis craccivora), which causes serious crop damage. To better understand cowpea and cowpea aphid interactions and to identify aphid effectors, proteins secreted into plant tissues to alter plant immunity, it is necessary to express these proteins in cowpea for functional analysis. Although a number of genetic and genomics resources for breeding cowpea have been developed, no functional genomics tool or transformation system exists for cowpea. Transiently expressing proteins by Agrobacterium tumefaciens is typically used for functional characterization of proteins in plants and for evaluating their effects on pathogens and pests. We therefore developed a transient expression system for cowpea by screening seven different A. tumefaciens strains for the expression of the transgene green fluorescent protein (GFP) tagged for nuclear localization. As a positive control for GFP expression, the GFP construct in the different A. tumefaciens strains was infiltrated, with a needless syringe, into leaves of Nicotiana benthamiana, a plant commonly used for transient expression. GFP expression in cowpea and N. benthamiana leaf nuclei was monitored using confocal microscopy. Our results indicate that all seven A. tumefaciens strains were able to transiently express GFP with at least a medium level of expression in 10-day-old cowpea cotyledons, but only some strains were able to transiently express GFP in cowpea 19-day-old cotyledons and trifoliate leaves. A. tumefaciens strain AGL01 appears to be the best candidate for use in transient expression in cowpea because it was the only strain that transiently expressed GFP at a high level in both 10-day-old cotyledons and trifoliate leaves.

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Nov 17th, 8:30 AM Nov 17th, 10:30 AM

Developing a Functional Genomics Tool for Cowpea

CREVELING 118

The cowpea plant (Vigna unguiculata) is a major source of nutrition in drought-ridden areas such as Sub-Saharan Africa. Like many crops, cowpea is plagued by numerous pests, one of which is the cowpea aphid (Aphis craccivora), which causes serious crop damage. To better understand cowpea and cowpea aphid interactions and to identify aphid effectors, proteins secreted into plant tissues to alter plant immunity, it is necessary to express these proteins in cowpea for functional analysis. Although a number of genetic and genomics resources for breeding cowpea have been developed, no functional genomics tool or transformation system exists for cowpea. Transiently expressing proteins by Agrobacterium tumefaciens is typically used for functional characterization of proteins in plants and for evaluating their effects on pathogens and pests. We therefore developed a transient expression system for cowpea by screening seven different A. tumefaciens strains for the expression of the transgene green fluorescent protein (GFP) tagged for nuclear localization. As a positive control for GFP expression, the GFP construct in the different A. tumefaciens strains was infiltrated, with a needless syringe, into leaves of Nicotiana benthamiana, a plant commonly used for transient expression. GFP expression in cowpea and N. benthamiana leaf nuclei was monitored using confocal microscopy. Our results indicate that all seven A. tumefaciens strains were able to transiently express GFP with at least a medium level of expression in 10-day-old cowpea cotyledons, but only some strains were able to transiently express GFP in cowpea 19-day-old cotyledons and trifoliate leaves. A. tumefaciens strain AGL01 appears to be the best candidate for use in transient expression in cowpea because it was the only strain that transiently expressed GFP at a high level in both 10-day-old cotyledons and trifoliate leaves.