Monica Schmidt

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Scholarly Contributions

Chapters

• Schmidt, M., & Frederico, M. L. (2016). Carotenoid biosynethesis in seeds: modern breeding and biotechnological approaches to enhance carotenoid accumulation. In Carotenoids in Nature; subcellular biochemistry.
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  MS and MLF co-corresponding authors
• Herman, E. M., Herman, E. M., & Schmidt, M. A. (2015). Towards Using Biotechnology to Modify Soybean Seeds as Protein Bioreactors. In Recent Advances in Gene Expression and Enabling Technologies in Crop Plants(pp 193-212). Springer New York. doi:10.1007/978-1-4939-2202-4_5
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  Soybean is an attractive platform as a protein bioreactor for the industrial scale-up of recombinant proteins. It is the major global vegetable protein commodity widely used as animal feed and as a component of processed food. The industrial capability to grow and process soybeans to obtain the protein fraction is a well-developed turnkey technology. There is an immense potential to create new feed products that meet the nutritional needs for production animals by developing soybeans as a protein production platform. Modified feed soybeans could be immediately fed into existing production paths as part of deployable products. The same technology used to modify soybeans to produce new feed and food proteins can be leveraged to produce other industrial proteins, where there is an increasing need for large-scale low-cost production for end uses such as enzymes for biofuel and food/feed processing. Producing proteins in soybean has the further advantage that soy oil is a global commodity coproduct with sufficient value to subsidize the costs of protein production. While the production of proteins in soybean is based on standard biotechnology approaches that have been developed for other plant species, soybean possesses some possibly unique intrinsic advantages that separate it from other plant protein bioreactor platforms, enhancing the capacity of soybean to stably accumulate foreign proteins. This chapter reviews the current state of engineering soybeans for protein production and outlines both the technical and biological aspects of protein production, and emerging opportunities to meet the product needs of the future.
• Hermand, E., & Schmidt, M. (2012). recent advances in plant expression in crop plants. In Towards using biotechnology to modify soybean seeds as protein bioreactors.
• Semenyuk, E., Schmidt, M. A., Woodford-thomas, T., Moravec, T., Moravec, T., & Semenyuk, E. G. (2015). Engineering Seeds for the Production and Delivery of Oral Vaccines. In Modification of seed composition to promote health nutrition, volume 51. John Wiley & Sons, Ltd. doi:10.2134/AGRONMONOGR51.C6
• Herman, E. M., Herman, E. M., & Schmidt, M. A. (2012). The Path to Economically Viable Foreign Protein Co-Products of Oilseeds. In Designing Soybeans for the 21st Century Markets. AOCS Press. doi:10.1016/B978-0-9830791-0-1.50016-9
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  Publisher Summary Biotechnology would play a fundamental role in implementing societal objectives, such as supplementing the U.S. and world energy supplies with soybean oil as a primary feedstock for biodiesel fuel. However, engineering soybean to produce more oil might not be sustainable unless value also is added to soybean co-products like soybean meal. A successful strategy for adding more value to soybean meal must consider the impact that more specialized high-value industrial protein products might exert upon critical applications such as commodity scale uses of soybean for feed and food. While the natural intrinsic characteristics of soybean protein already have significant value as a co-product, the strength of those advantages may be amplified through precise changes in the composition of soybean seed protein. This chapter discusses the need to understand the fundamental aspects of soybean molecular biology in efforts to reconstruct the protein composition in soybean seed and to translate this biology into useful tools for genetic engineering of novel beneficial traits in seeds.
• Schmidt, M. A., Dietrich, C. R., & Cahoon, E. B. (2005). Biotechnological Enhancement of Soybean Oil for Lubricant Applications. In synthetic, mineral oils and bio-based lubricants. CRC Press. doi:10.1201/9781420027181-26

Journals/Publications

• Schmidt, M. A., Herman, E. M., Kinney, A., He, Y., Samadar, P. P., Booth, R., Everard, J. D., Frank, M., Molloy, L., Abbitt, S., Coy, M., Liu, Z., Collet, K. H., & Shen, B. (2022). RNAi and CRISPR–Cas silencing E3-RING ubiquitin ligase AIP2 enhances soybean seed protein content. Journal of Experimental Botany. doi:10.1093/jxb/erac376
• Shen, B., Schmidt, M., Collet, K., Liu, Z., Coy, M., Abbitt, S., Molloy, L., Frank, M., Everard, J., Booth, R., Samadar, P., He, Y., Kinney, A., & Herman, E. M. (2022). RNAi and CRISPR-Cas silencing E3-ring ubiquitin ligase AiP2 enhances soybean seed protein content. Journal of Experimental Botany, 73(22), 7285-7297.
• Schmidt, M., Mao, Y., Opoku, J., & Mehl, H. (2021). Enzymatic degradation is an effective means to reduce aflatoxin contamination in maize. BMC Biotechnology, 21(1), 1-10.
• Konda, A., Nazarenus, T., Nguyen, H., Yang, J., Gelli, M., Swenson, S., Shipp, J., Schmidt, M., Cahoon, R., Ciftci, O., Zhang, C., Clemente, T., & Cahoon, E. (2020). Metabolic engineering of soybean seeds for enhanced vitamin E tocochromanol content and effects on oil antioxidant properties in polyunsaturated fatty acid rich germplasmi. Metabolic Engineering, 57, 63-73. doi:doi.org/10.1016/j.ymben.2019.10.005
• Krogdahl, A., Kortner, T., Gamil, A., Chikwati, E., Schmidt, M., Herman, E. M., Hymowitz, T., Teimouri, S., Kandel, S., & Storebakken, T. (2020). Soybean meal without Kunitz trypsin inhibitor, lectin and allergen P34/Gly m Bd 30k as protein source in diets for Atlantic salmon (Salmo salar, L) induce enteritis. Aquaculture.
• Herman, E. M., Schmidt, M., Radcliffe, J. S., Brito, L. F., Reddivari, L., & Schinckel, A. P. (2019). A swine model of soy protein–induced food allergenicity: implications in human and swine nutrition. Animal Frontiers, 9(3), 52-59. doi:10.1093/af/vfz025
• Krogdahl, A., Kortner, T., Gamil, A., Chikwati, E., Schmidt, M., Herman, E. M., Hymowitz, T., Teimouri, S., Kandel, S., & Storebakken, T. (2019). Soybean meal without Kunitz trypsin inhibitor, lectin and allergen P34/Gly m Bd 30k as protein source in diets for Atlantic salmon (Salmo salar, L) induce enteritis. Aquaculture.
• Radcliffe, J., Brito, L., Reddivari, L., Schmidt, M., Herman, E. M., & Schinckel, A. (2019). A swine model of soy protein-induced food allergenicity: implications in human and swine nutrition. Animal Frontiers, 9(3), 52-59.
• Barron, L. K., Bao, J. W., Aladeqbami, B. G., Schmidt, M., Herman, E. M., Erwin, C. R., & Warner, B. W. (2017). Soybean-derived human EGF enhances weight gain and lean muscle mass in malnurished mice. J Parenteral and Enteral Nutrition.
• Schmidt, M. A., & Herman, E. M. (2018). Characterization and functional biology of the soybean aleurone layer. BMC Plant Biology.
• Schmidt, M., & Pendarvis, K. (2017). Proteome rebalancing in Camelina occurs within the enlarged proteome induced by B-carotene accumulation. Transgenic Research.
• Thakare, D., Zhang, J., Cotty, P., Wing, R. A., & Schmidt, M. (2017). Host induced gene silencing inhibits aflatoxin production in transgenic maize when challenged with Aspergillus. Science Advances.
• Herman, E. M., & Schmidt, M. (2016). The potential of engineering functional-feed soybeans for sustainable aquaculture feed. Frontiers Plant Science, 7, 440. doi:Doi: 10.3389/fpls.2016.00440
• Herman, E. M., Schmidt, M., He, Y., Erwin, C., Guo, J., Sun, R., Pendarvis, K., & Warner, B. (2016). Transgenic soybean production of bioactive human epidermal growth factor (EGF). PLOS One. doi:DOI:10.1371/journal.pone.0157034
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  Necrotizing enterocolitis (NEC) is a devastating condition of premature infants that results from the gut microbiome invading immature intestinal tissues. This results in a life-threaten- ing disease that is frequently treated with the surgical removal of diseased and dead tis- sues. Epidermal growth factor (EGF), typically found in bodily fluids, such as amniotic fluid, salvia and mother’s breast milk, is an intestinotrophic growth factor and may reduce the onset of NEC in premature infants. We have produced human EGF in soybean seeds to lev- els biologically relevant and demonstrated its comparable activity to commercially available EGF. Transgenic soybean seeds expressing a seed-specific codon optimized gene encod- ing of the human EGF protein with an added ER signal tag at the N’ terminal were produced. Seven independent lines were grown to homozygous and found to accumulate a range of 6.7 +/- 3.1 to 129.0 +/- 36.7 μg EGF/g of dry soybean seed. Proteomic and immunoblot anal- ysis indicates that the inserted EGF is the same as the human EGF protein. Phosphoryla- tion and immunohistochemical assays on the EGF receptor in HeLa cells indicate the EGF protein produced in soybean seed is bioactive and comparable to commercially available human EGF. This work demonstrates the feasibility of using soybean seeds as a biofactory to produce therapeutic agents in a soymilk delivery platform.
• Schmidt, M. A., & Federico, M. L. (2016). Modern Breeding and Biotechnological Approaches to Enhance Carotenoid Accumulation in Seeds.. Sub-cellular biochemistry, 79, 345-58. doi:10.1007/978-3-319-39126-7_13
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  There is an increasing demand for carotenoids, which are fundamental components of the human diet, for example as precursors of vitamin A. Carotenoids are also potent antioxidants and their health benefits are becoming increasingly evident. Protective effects against prostate cancer and age-related macular degeneration have been proposed for lycopene and lutein/zeaxanthin, respectively. Additionally, β-carotene, astaxanthin and canthaxanthin are high-value carotenoids used by the food industry as feed supplements and colorants. The production and consumption of these carotenoids from natural sources, especially from seeds, constitutes an important step towards fortifying the diet of malnourished people in developing nations. Therefore, attempts to metabolically manipulate β-carotene production in plants have received global attention, especially after the generation of Golden Rice (Oryza sativa). The endosperms of Golden Rice seeds synthesize and accumulate large quantities of β-carotene (provitamin A), yielding a characteristic yellow color in the polished grains. Classical breeding efforts have also focused in the development of cultivars with elevated seed carotenoid content, with maize and other cereals leading the way. In this communication we will summarize transgenic efforts and modern breeding strategies to fortify various crop seeds with nutraceutical carotenoids.
• Schmidt, M. -., Hymowitz, T., & Herman, E. M. (2015). Breeding and characterization of soybean triple null; a stack of recessive alleles of Kunitz Trypsin Inhibitor, soybean Agglutinin and P34 allergen nulls. plant breeding.
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  EMH corresponding
• Schmidt, M. -., Parrott, W. A., Hildebrand, D. F., Berg, R. H., & Herman, E. M. (2015). transgenic soybean seeds accumulating b-carotene exhibit collateral enhancements of high oleate and high protein content traits. Plant Biotechnology.
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  MS corresponding and lead author
• Schmidt, M. A., Hymowitz, T., & Herman, E. M. (2015). Breeding and characterization of soybeanTriple Null;a stack of recessive alleles of Kunitz Trypsin Inhibitor, Soybean Agglutinin, and P34 allergen nulls. Plant Breeding, 134(3), 310-315. doi:10.1111/pbr.12265
• McCarthy, F., Schmidt, M. A., Parrott, W. A., Hildebrand, D. F., Berg, R. H., Cooksey, A., Pendarvis, K., He, Y., & Herman, E. M. (2014). Transgenic soya bean seeds accumulating β-carotene exhibit the collateral enhancements of oleate and protein content traits. Plant Biotechnology Journal, 13(4), 590-600. doi:10.1111/pbi.12286
• Schmidt, M. A., Barbazuk, B., Greg, M., Basil, N., M, S., W, H., & Eliot, H. M. (2011). Silencing of soybean seed storage proteins results in a rebalanced protein composition preserving seed protein content without major collateral changes in the metabolome and transcriptome. Plant Physiology, 156, 330-345.
• Schmidt, M., & Herman, E. (2010). Industial Protein Production Crops - New Needs and New Opportunities. GM Crops, 1, 2-7.
• Semenyuk, E. G., Schmidt, M. A., Beachy, R. N., Moravec, T., & Woodford-Thomas, T. (2010). Adaptation of an ecdysone-based genetic switch for transgene expression in soybean seeds. Transgenic Research, 19(6), 987-999.
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  PMID: 20191320;Abstract: Soybean was used as a model for studies of chemical induction of gene expression in seeds. A chimeric transcriptional activator, VGE, driven by the soybean seed glycinin G1 promoter, was used to induce the expression of an ER-targeted GFPKDEL reporter protein upon addition of the chemical ligand, methoxyfenozide. The chemical gene switch activated gene expression under in vitro conditions in somatic cotyledonary embryos and zygotic seed embryos cultured from transgenic soybean plants, as well as in seeds in planta under greenhouse conditions. The efficiency of induction of GFP expression under different growth conditions was strongly influenced by the developmental stage of the seed and availability of the inducer. The formation of ER-derived GFP-containing protein bodies in seed storage parenchyma cells was correlated with the level of induced expression. © 2010 Springer Science+Business Media B.V.
• Schmidt, M. A., LaFayette, P. R., Artelt, B. A., & Parrott, W. A. (2008). A comparison of strategies for transformation with multiple genes via microprojectile-mediated bombardment. In Vitro Cellular and Developmental Biology - Plant, 44(3), 162-168.
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  Abstract: The stable insertion and expression of multiple transgenes in crops is highly desirable, as the manipulation of complex agronomic traits and the introduction of novel biosynthetic pathways are dependent upon it. This study was performed to explore the frequency and efficiency of introducing multiple genes in soybean by using somatic embryogenesis and microprojectile bombardment transformation. The co-transformation frequency of six selectable marker or reporter genes (GusA, bleomycin resistance, glufosinate resistance, hygromycin resistance, green fluorescent protein, and kanamycin resistance) were followed throughout the T0, T1, and T2 generations. Three bombardment strategies were compared to determine the best method to generate transgenic plants that express the introduced transgenes and have a simple insertion pattern that would facilitate any downstream breeding. The plasmid bombardment treatments were (1) a six-gene-containing plasmid, (2) an equimolar treatment of five individual plasmids that collectively contained the six transgenes of interest (genes of glufosinate and hygromycin resistance were on the same plasmid), and (3) a 1:9 ratio mixture of the five plasmids, in which the plasmid containing the selectable marker used in the regeneration process, hygromycin resistance, was used in ninefold excess to all the other plasmids. Of the six bombardments performed per plasmid treatment, the results of seven independent events for the six-gene plasmid, four events for the 1:9 treatment, and a single regenerated event for the equimolar treatment indicate that containing all the transgenes on one plasmid just had an advantage in terms of frequency of a successful transformation events. Based on Southern analysis, the only events that contained all six transgenes was the one obtained by the equimolar treatment. No event was obtained that expressed all six transgenes, and certain transgenes seem to be non-randomly lost, namely gusA, bleomycin resistance, and glufosinate resistance, regardless of treatment. The addition of elements to optimize the expression of each gene cassette when multiple genes are in close proximity needs to be further investigated. © 2007 The Society for In Vitro Biology.
• Schmidt, M. a., & Herman, E. m. (2008). A RNAi knockdown of soybean 24 kDa oleosin results in the formation of micro oil bodies that aggregate to form large complexes of oil bodies and ER containing caleosin. Molecular Plant, 1, 910-924.
• Schmidt, M. a., & Herman, E. m. (2008). The collateral protein compensation mechanism can be exploited to enhance foreign protein accumulation in soybean seeds. Plant Biotechnology Journal, 6, 832-842.
• Livingston, D., Beilinson, V., Marina, K., Schmidt, M. a., Herman, E. m., & nielson, N. c. (2007). Reduction of protease inhibitor activity by expression of a mutant Bowman Birk gene in soybean seed. Plant Molecular Biology, 64, 397-408.
• Moravec, T., Schmidt, M. a., Herman, E. m., & Woodford Thomas, T. (2007). Production of Escherichia coli: heat labile toxin (Lt) B subunit in soybean seed and analysis of its immunogenicity. vaccine, 25, 1637-1657.
• Srivastava, V., Boyko, A., Kovalchuk, I., Sadder, M. T., Quain, M. D., Xie, C., Wen, J., Tucker, D., Tsay, H., Tapping, R. I., Stewart, C. N., Staats, E., Srivastava, V., Singer, S., Shen, L., Scoffield, J., Scoffield, J., Schmidt, M. A., Sadder, M. T., , Rosner, A., et al. (2007). 2007 In Vitro Biology meeting 2007 meeting of the Society for In Vitro Biology June 9–13, Indianapolis, IN. In Vitro Cellular & Developmental Biology – Plant, 43(6), 666-674. doi:10.1007/s11627-007-9069-y
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  Toll-like receptors (TLRs) constitute an essential family of pattern recognition molecules that, through the direct recognition of conserved microbial components, initiate inflammatory responses after infection. Phylogenetic evidence suggests that vertebrate TLRs are under strong purifying selection for the maintenance of function. Our lab is focused on a related group of vertebrate TLRs that comprise the TLR2 subfamily. The most closely related members, TLRs 1 and 6, appear to have arisen from a recent gene duplication event and have acquired differential microbial recognition specificity. We have characterized two single nucleotide polymorphisms (SNPs), P315L, and I602S, in human TLR1 that effect receptor function through different mechanisms. The 315L variant is associated with deficient recognition of microbial products whereas the 602S variant is associated with aberrant trafficking of the receptor to the cell surface. It is surprising to note that the 602S allele is associated with a decreased incidence of leprosy suggesting that Mycobacterium leprae subverts the TLR system as a mechanism of immune evasion. TLR1 I602S exhibits strikingly different allele frequencies among different races suggesting that if strong purifying selection took place, it was restricted by either additional genetic or environmental factors that were geographically constrained.High expression of transgenes is desired for molecular farming. However, transgenes are often subjected to gene silencing pathways in plant cells. Gene silencing may be triggered by the production of aberrant (hairpin) RNA molecule from a complex integration locus in plant genome or by the overexpression of transgene. Excessively transcribed RNA are subject to gene silencing even if they are produced from a single-copy locus. Therefore, we sought to determine how highly a transgene can be expressed before its transcript is subjected to gene silencing. In a previous study, we demonstrated that precise single-copy locus generated by Cre/lox-mediated site-specific integration (SSI) in rice is stably expressed at predictable levels through subsequent generations, and that its expression invariably doubled in homozygous progenies. To further explore the stability of the SSI locus and determine the expression-threshold of rice genomic sites, we generated transformation vectors containing 1–3 copies of 35S-GUS and 35S-GFP transgenes. These vectors were used to develop SSI locus containing 1–3 copies of each transgene in two different varieties of rice, Nipponbare and Taipei 309. SSI lines will be analyzed by PCR and Southern blotting to ascertain the presence of precise integration structures consisting of 1–3 copies of each trans-gene. The precise SSI lines will be subjected to quantitative GUS and GFP assay to determine if gene expression indeed increased with the increase in gene dosage. Molecular and expression data of SSI lines will be presented.Tobacco plants (Nicotiana tabacum L.) were transformed with a construct based on pCAMBIA 2301 containing a “hairpin” inverted repeat of 598 nucleotides derived from the Potato Virus Y (PVY) replicase (NIb gene) of the N strain (Robaglia et al., J. Gen. Virol. 70, 935, 1989). Such constructs confer virus resistance by a post translational gene silencing mechanism. Homozygous (T3) plants were challenged with a range of PVY strains and resistance was measured by symptom expression, ELISA titer, and back inoculation of controls with extracts from resistant plants. The nucleotide homology of PVY strains to the transgene was: WP (99.5%) PVY-NTN (96.3), PVY-H (95.6%), PVY-O (88.9%), strain 52 (88.3%), and local field isolates from tomato (86.8%), and pepper (86.3%). A transgenic tobacco line was immune to the five PVY strains with which the transgene had the greatest homology (WP, NTN, H, O, 52). Infection with the PVY isolates from tomato and pepper, which had the lowest degree of homology with the transgene, caused delayed symptom appearan
• Joseph, L., Hymowitz, T., Schmidt, M., & Herman, E. M. (2006). Evaluation of Glycine Germplasm for Nulls of the Immunodominant Allergen P34/Gly m Bd 30k. Crop Sciences, 46(4), 155-1763.
• Schmidt, M. A., Tucker, D. M., Cahoon, E. B., & Parrott, W. A. (2005). Towards normalization of soybean somatic embryo maturation. Plant Cell Reports, 24(7), 383-391.
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  PMID: 15856235;Abstract: Soybean (Glycine max L. Merrill) somatic embryos have been useful for assaying seed-specific traits prior to plant recovery. Such traits could be assessed more accurately if somatic embryos more closely mimicked seed development. Amino acid supplements, carbon source, and abscisic acid and basal salt formulations were tested in an effort to modify existing soybean embryogenesis histodifferentiation/maturation media to further normalize the development of soybean somatic embryos. The resultant liquid medium, referred to as soybean histodifferentiation and maturation medium (SHaM), consists of FNL basal salts, 3% sucrose, 3% sorbitol, filter-sterilized 30 mM glutamine and 1 mM methionine. SHaM-derived somatic embryos are more similar to seed in terms of protein and fatty acid/lipid composition, and conversion ability, than somatic embryos obtained from traditional soybean histodifferentiation and maturation media. © Springer-Verlag 2005.
• Schmidt, M. A., Martin, G. S., Artelt, B. J., & Parrott, W. A. (2004). Increased transgene expression by breeding and selection in white clover. Crop Science, 44(3), 963-967.
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  Abstract: To determine if standard breeding methodology is applicable to transgenes, phenotypic recurrent selection was used to select for increased transgene expression in white clover, Trifolium repens L. Plants were transformed with nptII and gusA, and selected on 100 mg L-1 of kanamycin. Independently transformed plants were intercrossed, and the progeny was germinated on 200, 300, or 400 mg L-1 of kanamycin. Those seedlings surviving on 400 mg-1 were in turn intercrossed, and the progeny was selected on 300, 400, or 500 mg L-1 of kanamycin. NPTII levels were measured in each selected population, and Southern blots were made from individuals in each population. The highest-expressing individual in the T 2 had levels of NPTII that were more than four times higher than those in the highest parent. With selection on increasing levels of kanamycin, average expression across each generation went from 0.033 ng μg-1 NPTII in the parents to 0.095 ng μg-1 in the selected T 1 plants to 0.539 ng μg-1 in the selected T2 plants. Southern hybridization suggested that plants displaying a heightened level of nptII expression in the T1 and T2 fell into two categories. The first contained one particular transgenic event, implicating the importance of other genomic factors in modulating gene expression. Alternatively, the plants had an accumulation of various nptII loci, suggesting an association between multiple transgene copies and high expression levels. On the basis of these results, selection for transgene expression appears to be a viable option for plant breeding programs.
• Schmidt, M. a., & Herman, E. m. (2004). Endoplasmic reticulum to vacule trafficking of endoplasmic reticulum bodies provides an alternative pathway for protein transfer to teh vacuole. Plant Physiology, 136, 3440-3446.
• Thomson, J. M., Lafayette, P. R., Schmidt, M. A., & Parrott, W. A. (2002). Artificial gene-clusters engineered into plants using a vector system based on intron- and intein-encoded endonucleases. In Vitro Cellular and Developmental Biology - Plant, 38(6), 537-542.
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  Abstract: The ability to create artificial gene-clusters for genetic transformation could facilitate the development of crops with multiple engineered traits, or with traits which result from the expression of multiple genes. A simple method to assemble artificial gene-clusters was developed by designing a multiple cloning site consisting of an array of homing endonuclease cleavage sites into a single vector. These enzymes are also known as intron- or intein-encoded endonucleases, and have very long recognition sequences, which makes them very rare cutters. The resulting vectors are pUGA for microprojectile-mediated transformation, and pUGA2 for Agrobacterium-mediated transformation. In addition, a series of unidirectional shuttle vectors containing various combinations of homing endonuclease restriction sites was constructed. Gene cassettes can be cloned into individual shuttles, and then transferred to either pUGA or pUGA2 to construct artificial gene-clusters. To test the feasibility of this approach, a six-gene cluster was constructed and transformed into soybean via microprojectile bombardment and into tobacco via Agrobacterium. The genes were assayed for expression in both the T0 and T1 generations for three independent transgenics. Up to five of the six genes were expressed. Additional changes to the construction of individual gene cassettes may improve the frequency with which all genes in the cluster are expressed.
• Thomson, J. M., Schmidt, M. A., Parrott, W. A., & Lafayette, P. R. (2002). ARTIFICIAL GENE-CLUSTERS ENGINEERED INTO PLANTS USING A VECTOR SYSTEM BASED ON INTRON- AND INTEIN-ENCODED ENDONUCLEASES. In Vitro Cellular & Developmental Biology – Plant, 38(6), 537-542. doi:10.1079/ivp2002329
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  The ability to create artificial gene-clusters for genetic transformation could facilitate the development of crops with multiple engineered traist, or with traits which result from the expression of multiple genes. A simple method to assemble artificial gene-clusters was developed by designing a multiple cloning site consisting of an array of homing endonuclease cleavage sites into a single vector. These enzymes are also known as intron-or intein-encoded endonucleases, and have very long recognition sequences, which makes them very rare cutters. The resulting vectors are pUGA for microprojectile-mediated transformation, and pUGA2 for Agrobacterium-mediated transformation. In addition, a series of unidirectional shuttle vectors containing various combinations of homing endonuclease restriction sites was constructed. Gene cassettes can be cloned into individual shuttles, and then transferred to either pUGA or pUGA2 to construct artificial gene-clusters. To test the feasibility of this approach, a six-gene cluster was constructed and transformed into soybean via microprojectile bombardment and into tobacco via Agrobacterium. The genes were assayed for expression in both the T0 and T1 generations for three independent transgenics. Up to five of the six genes were expressed. Additional changes to the construction of individual gene cassettes may improve the frequency with which all genes in the cluster are expressed.
• Schmidt, M. A., & Parrott, W. A. (2001). Quantitative detection of transgenes in soybean [Glycine max (L.) Merrill] and peanut (Arachis hypogaea L.) by real-time polymerase chain reaction. Plant Cell Reports, 20(5), 422-428.
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  Abstract: Quantitative real-time polymerase chain reaction (PCR) assays were designed that enabled the zygosity of transgenes in soybean [Glycine max (L.) Merrill] and peanut (Arachis hypogaea L.) to be determined. The two zygosity assays, based on TaqMan technology that uses a fluorogenic probe which hybridizes to a PCR target sequence flanked by primers, were both accurate and reproducible in the determination of the number of transgenes present in a cell line. In the first assay, in which TaqMan assays were performed on increasing amounts of a plasmid containing the transgene of interest, a linear relationship between the level of fluorescence and the template amount was produced. Using the resultant linear relationships as standard curves, we were able to determine the zygosity of both soybeans segregating for the cry1Ac transgene and that of a T1 peanut segregating for the hph transgene. In the second assay, a relative determination of copy number (referred to as comparative Ct) was performed on transgenic soybeans by comparing the amplification efficiency of the transgene of interest to that of an endogenous gene in a multiplexed PCR reaction. Both methods proved to be sufficiently sensitive to differentiate between homozygotes and hemizygotes. These assays have numerous potential applications in plant genetic engineering and tissue culture, including the hastening of the identification of transgenic tissue, selecting transformation events with a low number of transgenes and the monitoring of the transmission of transgenes in subsequent crosses.
• Lovett Doust, J., Schmidt, M., & Lovett Doust, L. (1994). Biological Assessment of aquatic pollution: a review with emphasis on plants as biomonitors. Biological Reviews, 69, 147-186.
• Doust, L. L., Doust, J. L., & Schmidt, M. (1993). In praise of plants as biomonitors - Send in the clones. Functional Ecology, 7(6), 754-758.

Presentations

• Schmidt, M. (2021, June). Engineering Functional Foods. CN Yang Scholars Program. virtual: Oregon State University AND University of Singamore.
• Schmidt, M. (2021, Nov). High resolution soybean genome of high-protein mutants. PACBIO Seminar -- Better Data Yields Better Biology. Arizona Genomics Institute.
• Schmidt, M. (2019, January). Enhanced accumulation of B-carotene in seeds of both soybean and camellia result in altered seed composition. PAG 2019. San Diego.
• Schmidt, M. (2018, April). Functional Foods Through Biotechnology. University of Florida.
• Schmidt, M. (2018, Jan). HIGS as an effective means to alleviate aflatoxin contamination. PAG 2018. San Diego, CA USA.
• Schmidt, M. (2018, June). HIGS to eliminate mycotoxins. Corn Utilization and Technology meeting. St.Louis MO USA.
• Schmidt, M. (2018, March). Aflatoxin-free transgenic maize using host-induced gene silencing. World Mycotoxin Meeting. Amsterdam, Netherlands.
• Schmidt, M. (2018, Nov). Characterization of aleurone layer in soybean. Crop Science Society. Baltimore MD USA.
• Schmidt, M. (2018, Sept). Functional Foods through Biotechnology. Montana State University. Bozeman MT USA.
• Schmidt, M. (2018, Sept). Functional Foods through Biotechnology. University of California Davis. Davis, CA USA.
• Schmidt, M. (2017, Dec). Aflatoxin-free maize through the use of host-induced gene silencing. American Seed Trade Association (CSS). Chicago IL: American Seed Trade.
• Schmidt, M. (2017, July). Aflatoxin-free crops through the use of biotechnology. Opportunities for Reduction of Aflatoxin Contamination of Food. Cold Spring Harbor NY.
• Schmidt, M. (2017, March). Functional Foods Through Biotechnology. Science City. Tucson AZ: Tucson Festival of Books.
• Schmidt, M. (2017, May). assessing phenotypic variation in soybean seed protein traits using GFP as a reporter in both mutagenesis and transgenomic approaches. Biotechnology Risk Assessment. Washington DC: NIFA.
• Schmidt, M. (2016, April). Functional Foods through Biotechnology. Nutritional Sciences seminar series. University of Arizona.
• Schmidt, M. (2016, Feb). Functional Foods -- using biotechnology to engineer healthier foods/feeds for consumers. Indian Institute of Technology and Science and Technology Institute. Guwahati and Jorhat India: Borlaug Fellowship.
• Schmidt, M. (2016, June). Functional Foods -- making food/feed better through biotechnology. World Congress on In Vitro Biology. San Diego CA USA.
• Schmidt, M. (2016, May). Proteome rebalancing in transgenic Camelina occurs within the enlarged proteome induced by B-carotene accumulation and storage protein suppression. Gordon research conference -- carotenoid biosynthesis, functions and applications to human health. Tuscany, Italy.
• Schmidt, M. (2016, Nov). Aflatoxin-free transgenic maize plants vis HIGS. American Society of Agronomy/ Crop Science Society. Phoenix AZ USA.
• Schmidt, M. (2016, jan). Modern Crops Through Breeding and Genetic Engineering. City HighSchool. City High School Tucson AZ.
• He, Y., Schmidt, M. a., Sun, R., Erwin, C., Warner, B., & Herman, E. (2015, May). Production of Transgenic soybean seeds expressing human epidermal growth factor (hEGF) for a therapeutic formula to prevent neonatal necrotizing enterocolitis. SIVB Conference:Plant Biotechnology Section. Tucson AZ: society of in vitro biology.
• Thake, D., Cotty, P., & Schmidt, M. (2015, May). Aflatoxin free transgenic maize. SIVB Conference. Tucson AZ: Society of In Vitro Biology.

Poster Presentations

• Schmidt, M., & Herman, E. M. (2019, January). Altered expression of the ubiquitin E3 ring ligase AiP2 increases soybean seed protein content. PAG 2019. San Diego.
• Verla, M., Style, C., Stoll, B., Herman, E. M., Schmidt, M., Burrin, D., Yu, L., & Olutoye, O. (2019, March). Efficacy of Soybean-derived Epidermal Growth Factor Therapy in the Preterm Piglet Model of Necrotizing Enterocolitis. American College of Surgeons 2019 Clinical CongressAmerican College of Surgeons.
• Schmidt, M. (2018, May). Assessing genetic diversity through mutagenesis and transgenesis. BRAG NIFA meeting. Washington DC, USA.
• Schmidt, M. (2016, Aug). Increased protein content in transgenic seeds with enhanced b-carotene content. SOY2016- Molecular and Cellular Biology of the Soybean 16th Biennial Conference. Columbus, OH USA.
• Schmidt, M. -. (2012, October). epigenetic events in transgenic soybean plants leading to non-seed phenotypes. Legume Genomics Conference. Hyderabad India.