Deyu Xie

Plant lectins are a large group of proteins that do not have catalytic activities. They are carbohydrate-binding proteins that bind sugar groups of proteins, peptides, polysaccharides, lipids, and other glycoconjugates. However, the production of lectins in plants are not high. Under this project, we propose to test different Escherichia coli and expression vectors to induce plant lectins, develop expression induction approaches for a high production, and then establish a combination of tag and affinity-column chromatography methods to isolate lectins.

Borlaug Fellowship Program for Turkey fellow to spend 12 weeks at NC State conducting research to weaken wheat phenylalanine ammonia-lyase gene promotor by homology directed repaid CRISPR-Cas9 technology to obtain more economical and effective biofuel production.

Using antibiotics in feed has become a global concern due to its contribution to the antibiotic resistance. Exploring the alternatives to antibiotics is necessary for human health and animal productive efficiency. Proanthocyanidins extracted from engineered novel plants and cells with biological and medicinal activities will be screened in this project.

Borlaug Fellowship Program for Sri Lanka fellow to spend 12 weeks at NC State conducting research to identify and characterize candidate resistance genes against blister blight disease in teac (Chinese sinensis).

The fellow will work on a topic entitled ����������������Metabolomics of cacao��������������� under the mentoring of Dr. Deyu Xie. Through this project, the fellow will learn extraction approaches of metabolites from cacao nuts and chocolate, HPLC-qTOF-MS/MS based metabolomics, structural annotation of metabolites, principal component analysis, and heatmap and hierarchy clustering analysis. The fellow will be instructed to read important literatures to understand the principle of metabolomics. In addition, the fellow will attend Xie������������������s weekly lab meeting and departmental seminars to gain additional opportunities to learn research progresses in other topics. By the end of this program, the fellow will master main metabolomics-based technology for characterization of functional metabolites in cacao nuts and will be able to develop research projects at their home university.

1. Understanding downregulation mechanisms of tobacco alkaloid biosynthesis by PAP1 Specific goals include: ��������������� 1-1: Transcriptional analysis of known genes involved in tobacco alkaloid biosynthesis ��������������� 1-2: PAP1 interaction with promoters of down-regulated pathway genes ��������������� 1-3: PAP1 interaction with NtMYC2a, b, c ��������������� 1-4: Transcriptomics of red versus vector control transgenic plants as well as wild-type plants ��������������� 1-5: PAP1 or PAP1 complex-based biotechnology for reduction of nicotine, nornicotine and TSNA 2. Screening of anthocyanin-pigmentation from fast neutron deletion mutagenesis of K326 and Dark tobacco for red tobacco varieties

Variation in floral display (inflorescence) affects the success of plant reproduction and the yield of a crop by influencing seed number and dispersal/harvest ability. Despite its importance, little progress has been made in understanding how developmental and genetic changes have shaped inflorescence architectures in angiosperm evolution, in part because existing model organisms exhibit little variation in these traits. Species of Dogwood (Cornus L.) are popular ornamental trees in American landscapes due to their spectacular inflorescences often associated with large showy (petaloid) bracts. The genus offers us a unique opportunity to tackle this important problem. Cornus exhibits a wide variation in inflorescence structure, including heads, umbels and compound cymes. A recent breakthrough in our laboratory has resulted in successful regeneration and transformation of a key species of the genus. This new ability along with our recent achievements in phylogenetic reconstruction and nearly completed comparative developmental studies, now provides a timely opportunity to develop a model for investigating the molecular mechanisms shaping inflorescence architectures in a non-model plant lineage. In this proposal, we aim to test four hypotheses to gain insights into the changes in developmental and genetic mechanisms that may have led to alteration of inflorescence forms in dogwoods. Hypotheses: 1. Differences in early development processes lay the ground for divergence of inflorescence architectures in dogwood species. 2. Spatial, temporal, and quantitative variation of expression of conserved key inflorescence regulatory genes are essential for the changes of inflorescence architecture. 3. Expression of petal identity genes in bracts is essential for the origin of the petaloidy of bracts in dogwoods. 4. Spatial, temporal, and quantitative variation of expression of other genes are essential for the modification of inflorescence architecture and origin of petaloid bracts in dogwoods. Objectives: 1. Complete the comparative developmental characterization of four Cornus inflorescence types. 2. Comparative characterization of the expression of conserved key inflorescence regulators and petal identity genes in different inflorescence types. 3. Identify new genes regulating inflorescence development and bract petaloidy in dogwood. 4. Optimize the existing transformation system of Cornus canadensis and test the function of conserved key inflorescence regulatory genes Intellectual merits: This project represents the first study investigating the molecular controls of inflorescence development and evolution using a comparative approach on multiple closely related woody species from a non-model plant lineage. Although the proposal has technical challenges, we have overcome the most significant hurdles and achieving of our objectives would offer novel insights into the genetic basis underlying the evolutionary transitions of floral display strategies, of which the knowledge is presently lacking. The transformation system has tremendous potentials for future research of genetic controls of other plant traits including resistance to fungal diseases and drought, and flowering time divergence among species. By providing a transformation system in a non-model plant, this study will potentially provide a new tool for genetic analysis in other plants that are more closely related to the dogwoods than to the few existing model species. Broader impact: The project will not only enhance our understanding of inflorescence development and evolution in angiosperms, but also hold promise in breeding and bioengineering of dogwoods. Biotechnological improvement of Cornus species in display, disease and drought resistance holds tremendous industrial potential. Thus the results of this project will have broad interests to the scientific, biotechnolgical, and industrial communities. Perfection of the transformation system will immediately benefit the research of identifying genes resistant to the dogwood anthracnose disease that

The new equipment will serve a diverse user community. Because of its combined capability as a highthroughputb sequencer that generates reads ultra fast, it will be an essential tool in research requiring: 1) variant detection for genotyping breeding populations, 2) near real-time metagenomics remediation assays and other population surveys, 3) SNP detection, genotyping, and genetic mapping, 4) transcriptome sequencing and gene expression analysis, and 5) full genome and targeted resequencing (including ChIP seq and bisulfite-treated DNA applications)

Comparison of metabolomes in leaves of tobacco grown in India, Brazil, and North Carolina; Establishment of flavor-related metabolome; Identification of biosynthetic pathways regulated by biogeography-specific conditions; Establishment of flavor-specific network pathways to predict potential genes to direct next step of gene cloning and metabolic engineering

Fossil fuels (crude oil and natural gas) are currently the major energy source in the United States and many other countries. However, the reserves of crude oil and natural gas are limited and will be exhausted in the near future if their consumption continues at its current rate. Campbell and Laherrere used several different techniques to estimate the current known crude oil reserves and the reserves as yet undiscovered and concluded that the decline in world-wide crude oil production will begin before 2010. They also predicted that annual global oil production would decline from the current 25 billion barrels to around 5 billion barrels in 2050. Because the economy in the U.S. and many other nations depends on fossil fuels, the consequence of inadequate supply of crude oil and natural gas would be severe. Therefore, our government has realized that it is urgent to explore alternative renewable energy sources for sustainable economic development; on the other hand, there are a tremendous amount of biomass and organic wastes such as agricultural and forest residues, grasses, and trees that can be used for renewable energy production. Switchgrass is one of the top renewable plant resources to meet future requirement of energy. Certain extensive research has been conducted on conversion of switchgrass to ethanol over the past several years. However, most of the previous efforts have been mainly focused on processing of the switchgrass material, including the costly pretreatment to remove lignin. Genetic manipulation to reduce lignin content in lignocellulosic materials has been shown to be an efficient way to improve or eliminate pretreatment in other plants, but has not been applied to switchgrass. In this project, for the first time we propose to develop an integrated genetic technology to reduce lignin level and reduce the cost of processing for improved bioethanol production of switchgrass. The long term goal is to genetically develop new switchgrass with reduced levels of lignin and increased levels of cellulose, and to express enzymes which facilitate processing and reduce cost in efficient production of bioethanol. Underlying this particular goal, we have four specific aims for the proposed project: 1. Cloning of switchgrass HCT gene 2. Suppression of the lignin biosynthesis in switchgrass by suppressing expression of the HCT and 4CL genes 3. Expression of the thermostable OSS1949 cellulase in switchgrass to reduce cost during fermentation 4. Identify lignin-reduced and cellulase-expressing transgenic switchgrass plants as value-added feedstocks Our research team has years of experience in grass transformation, lignin/cellulose synthesis biochemistry and molecular biology, and metabolic engineering. We do not anticipate any major difficulties to achieve our aims.