Zionsville, IN, United States
Zionsville, IN, United States

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Damaj M.B.,Texas AgriLife Research Center | Beremand P.D.,Texas A&M University | Buenrostro-Nava M.T.,Texas AgriLife Research Center | Ivy J.,Texas A&M University | And 6 more authors.
Genome | Year: 2010

The availability of a wider range of promoters for regulated expression in valuable transgenic crops would benefit functional genomics studies and current biotechnology programs aimed at improved productivity. Polymerase chain reaction (PCR)-based genome walking techniques are commonly used to isolate promoters or 5′ flanking genomic regions adjacent to known cDNA sequences in genomes that are not yet completely sequenced. However, these techniques are problematic when applied directly to DNA isolated from crops with highly complex and large genomes. An adaptor ligationmediated PCR-based BAC genome walking method is described here for the efficient isolation of promoters of multigene family members, such as the putative defense and fiber biosynthesis DIRIGENT genes that are abundant in the stem of sugarcane, a species with a highly polyploid genome. The advantage of this method is the efficient and specific amplification of the target promoter using BAC genomic DNA as template for the adaptor ligation-mediated PCR walking.


Damaj M.B.,Texas AgriLife Research Center | Kumpatla S.P.,DowAgroSciences LLC | Emani C.,Texas A&M University | Beremand P.D.,Texas A&M University | And 6 more authors.
Planta | Year: 2010

Transcription profiling analysis identified Saccharum hybrid DIRIGENT (SHDIR16) and O-METHYLTRANSFERASE (SHOMT), putative defense and fiber biosynthesis-related genes that are highly expressed in the stem of sugarcane, a major sucrose accumulator and biomass producer. Promoters (Pro) of these genes were isolated and fused to the β-glucuronidase (GUS) reporter gene. Transient and stable transgene expression analyses showed that both ProDIR16:GUS and ProOMT:GUS retain the expression characteristics of their respective endogenous genes in sugarcane and function in orthologous monocot species, including rice, maize and sorghum. Furthermore, both promoters conferred stem-regulated expression, which was further enhanced in the stem and induced in the leaf and root by salicylic acid, jasmonic acid and methyl jasmonate, key regulators of biotic and abiotic stresses. ProDIR16 and ProOMT will enable functional gene analysis in monocots, and will facilitate engineering monocots for improved carbon metabolism, enhanced stress tolerance and bioenergy production. © Springer-Verlag 2010.


Shukla J.N.,University of Kentucky | Kalsi M.,University of Kentucky | Sethi A.,DuPont Pioneer | Narva K.E.,DowAgrosciences LLC | And 4 more authors.
RNA Biology | Year: 2016

RNA interference (RNAi) has become a widely used reverse genetic tool to study gene function in eukaryotic organisms and is being developed as a technology for insect pest management. The efficiency of RNAi varies among organisms. Insects from different orders also display differential efficiency of RNAi, ranging from highly efficient (coleopterans) to very low efficient (lepidopterans). We investigated the reasons for varying RNAi efficiency between lepidopteran and coleopteran cell lines and also between the Colorado potato beetle, Leptinotarsa decemlineata and tobacco budworm, Heliothis virescens. The dsRNA either injected or fed was degraded faster in H. virescens than in L. decemlineata. Both lepidopteran and coleopteran cell lines and tissues efficiently took up the dsRNA. Interestingly, the dsRNA administered to coleopteran cell lines and tissues was taken up and processed to siRNA whereas the dsRNA was taken up by lepidopteran cell lines and tissues but no siRNA was detected in the total RNA isolated from these cell lines and tissues. The data included in this paper showed that the degradation and intracellular transport of dsRNA are the major factors responsible for reduced RNAi efficiency in lepidopteran insects. © 2016 Taylor & Francis Group, LLC.


Ruf-Pachta E.K.,Kansas State University | Rule D.M.,Kansas State University | Rule D.M.,DowAgroSciences LLC | Dille J.A.,Kansas State University
Weed Science | Year: 2013

Palmer amaranth influences selection of crop production practices such as irrigation, nitrogen (N) application, and weed control. The objectives of this research were to determine if Palmer amaranth was more responsive to applied N than corn and if this differed under dryland and irrigated conditions in Kansas. Field experiments were conducted near Manhattan, KS, in 2005 and 2006 to evaluate the influence of N rate and Palmer amaranth densities when grown with corn in two soil moisture environments. A very drought-stressed environment and a well-watered environment occurred in 2006, while both environments in 2005 were intermediate. Dryland weed-free corn yields were 46.5% of irrigated corn yields at the high N rate across years. Irrigated corn yields responded to increasing N rates. In the presence of Palmer amaranth, parameter estimates I and A for the yield loss relationship were not different across N rates for each environment and year except 2006 where 100% yield loss was estimated in dryland compared to 62.5% loss in irrigated environment at high N rates. In three of four environment-years, N rate did not affect the corn yield loss relationship with weed density. In 2006 irrigated environment, greater N rates had less corn yield loss caused by Palmer amaranth. By corn anthesis, weed-free corn biomass was 167.5% greater in irrigated than dryland environments in 2006. Palmer amaranth with no corn increased its biomass by 373 and 361% as N rate increased in 2005 and 2006, respectively. Nitrogen concentrations in plant tissues of corn or weed increased similarly as N rates increased from 0 to 224 kg N ha -1, thus highlighting that both corn and Palmer amaranth responded similarly to increasing N. In general, soil moisture environment was most critical when determining potential corn yield, followed by Palmer amaranth density and N rate. Nomenclature: Palmer amaranth, Amaranthus palmeri S. Wats. AMAPA; corn, Zea mays L. "DKC60-19RR".© Weed Science Society of America 2013.

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