Manipulation of plants by transformation with sequences promoting cell division

Polynucleotides encode polypeptides for increasing the rate of growth of plants. Introduction of the polynucleotides into plants produces plants having altered characteristics, such as increased growth, increased leaf area and reduced fertility. Expression of polypeptides in plants or plant cells promotes cell division. Expression of the polynucleotides in plants in the antisense orientation produces plants that are sterile or have smaller leaves.https://digitalcommons.mtu.edu/patents/1012/thumbnail.jp

Genome-wide transcriptome analysis of the transition from primary to secondary stem development in Populus trichocarpa

<p>Abstract</p> <p>Background</p> <p>With its genome sequence and other experimental attributes, <it>Populus trichocarpa </it>has become the model species for genomic studies of wood development. Wood is derived from secondary growth of tree stems, and begins with the development of a ring of vascular cambium in the young developing stem. The terminal region of the developing shoot provides a steep developmental gradient from primary to secondary growth that facilitates identification of genes that play specialized functions during each of these phases of growth.</p> <p>Results</p> <p>Using a genomic microarray representing the majority of the transcriptome, we profiled gene expression in stem segments that spanned primary to secondary growth. We found 3,016 genes that were differentially expressed during stem development (Q-value ≤ 0.05; >2-fold expression variation), and 15% of these genes encode proteins with no significant identities to known genes. We identified all gene family members putatively involved in secondary growth for carbohydrate active enzymes, tubulins, actins, actin depolymerizing factors, fasciclin-like AGPs, and vascular development-associated transcription factors. Almost 70% of expressed transcription factors were upregulated during the transition to secondary growth. The primary shoot elongation region of the stem contained specific carbohydrate active enzyme and expansin family members that are likely to function in primary cell wall synthesis and modification. Genes involved in plant defense and protective functions were also dominant in the primary growth region.</p> <p>Conclusion</p> <p>Our results describe the global patterns of gene expression that occur during the transition from primary to secondary stem growth. We were able to identify three major patterns of gene expression and over-represented gene ontology categories during stem development. The new regulatory factors and cell wall biogenesis genes that we identified provide candidate genes for further functional characterization, as well as new tools for molecular breeding and biotechnology aimed at improvement of tree growth rate, crown form, and wood quality.</p

A feasibility study of hand kinematics for EVA analysis using magnetic resonance imaging

A new method of analyzing the kinematics of joint motion is developed. Magnetic Resonance Imaging (MRI) offers several distinct advantages. Past methods of studying anatomic joint motion have usually centered on four approaches. These methods are x-ray projection, goniometric linkage analysis, sonic digitization, and landmark measurement of photogrammetry. Of these four, only x-ray is applicable for in vivo studies. The remaining three methods utilize other types of projections of inter-joint measurements, which can cause various types of error. MRI offers accuracy in measurement due to its tomographic nature (as opposed to projection) without the problems associated with x-ray dosage. Once the data acquisition of MR images was complete, the images were processed using a 3D volume rendering workstation. The metacarpalphalangeal (MCP) joint of the left index finger was selected and reconstructed into a three-dimensional graphic display. From the reconstructed volumetric images, measurements of the angles of movement of the applicable bones were obtained and processed by analyzing the screw motion of the MCP joint. Landmark positions were chosen at distinctive locations of the joint at fixed image threshold intensity levels to ensure repeatability. The primarily two dimensional planar motion of this joint was then studied using a method of constructing coordinate systems using three (or more) points. A transformation matrix based on a world coordinate system described the location and orientation of a local target coordinate system. Future research involving volume rendering of MRI data focusing on the internal kinematics of the hand's individual ligaments, cartilage, tendons, etc. will follow. Its findings will show the applicability of MRI to joint kinematics for gaining further knowledge of the hand-glove (power assisted) design for extravehicular activity (EVA)

Validating internal controls for quantitative plant gene expression studies

BACKGROUND: Real-time reverse transcription PCR (RT-PCR) has greatly improved the ease and sensitivity of quantitative gene expression studies. However, accurate measurement of gene expression with this method relies on the choice of a valid reference for data normalization. Studies rarely verify that gene expression levels for reference genes are adequately consistent among the samples used, nor compare alternative genes to assess which are most reliable for the experimental conditions analyzed. RESULTS: Using real-time RT-PCR to study the expression of 10 poplar (genus Populus) housekeeping genes, we demonstrate a simple method for determining the degree of stability of gene expression over a set of experimental conditions. Based on a traditional method for analyzing the stability of varieties in plant breeding, it defines measures of gene expression stability from analysis of variance (ANOVA) and linear regression. We found that the potential internal control genes differed widely in their expression stability over the different tissues, developmental stages and environmental conditions studied. CONCLUSION: Our results support that quantitative comparisons of candidate reference genes are an important part of real-time RT-PCR studies that seek to precisely evaluate variation in gene expression. The method we demonstrated facilitates statistical and graphical evaluation of gene expression stability. Selection of the best reference gene for a given set of experimental conditions should enable detection of biologically significant changes in gene expression that are too small to be revealed by less precise methods, or when highly variable reference genes are unknowingly used in real-time RT-PCR experiments

Genome scale transcriptome analysis of shoot organogenesis in Populus

<p>Abstract</p> <p>Background</p> <p>Our aim is to improve knowledge of gene regulatory circuits important to dedifferentiation, redifferentiation, and adventitious meristem organization during <it>in vitro </it>regeneration of plants. Regeneration of transgenic cells remains a major obstacle to research and commercial deployment of most taxa of transgenic plants, and woody species are particularly recalcitrant. The model woody species <it>Populus</it>, due to its genome sequence and amenability to <it>in vitro </it>manipulation, is an excellent species for study in this area. The genes recognized may help to guide the development of new tools for improving the efficiency of plant regeneration and transformation.</p> <p>Results</p> <p>We analyzed gene expression during poplar <it>in vitro </it>dedifferentiation and shoot regeneration using an Affymetrix array representing over 56,000 poplar transcripts. We focused on callus induction and shoot formation, thus we sampled RNAs from tissues: prior to callus induction, 3 days and 15 days after callus induction, and 3 days and 8 days after the start of shoot induction. We used a female hybrid white poplar clone (INRA 717-1 B4, <b><it>Populus tremula × P. alba</it></b>) that is used widely as a model transgenic genotype. Approximately 15% of the monitored genes were significantly up-or down-regulated when controlling the false discovery rate (FDR) at 0.01; over 3,000 genes had a 5-fold or greater change in expression. We found a large initial change in expression after the beginning of hormone treatment (at the earliest stage of callus induction), and then a much smaller number of additional differentially expressed genes at subsequent regeneration stages. A total of 588 transcription factors that were distributed in 45 gene families were differentially regulated. Genes that showed strong differential expression included components of auxin and cytokinin signaling, selected cell division genes, and genes related to plastid development and photosynthesis. When compared with data on in vitro callogenesis in <it>Arabidopsis</it>, 25% (1,260) of up-regulated and 22% (748) of down-regulated genes were in common with the genes regulated in poplar during callus induction.</p> <p>Conclusion</p> <p>The major regulatory events during plant cell organogenesis occur at early stages of dedifferentiation. The regulatory circuits reflect the combinational effects of transcriptional control and hormone signaling, and associated changes in light environment imposed during dedifferentiation.</p

EVA Glove Research Team

The goal of the basic research portion of the extravehicular activity (EVA) glove research program is to gain a greater understanding of the kinematics of the hand, the characteristics of the pressurized EVA glove, and the interaction of the two. Examination of the literature showed that there existed no acceptable, non-invasive method of obtaining accurate biomechanical data on the hand. For this reason a project was initiated to develop magnetic resonance imaging as a tool for biomechanical data acquisition and visualization. Literature reviews also revealed a lack of practical modeling methods for fabric structures, so a basic science research program was also initiated in this area