Browsing by Author "Nowak, Jerzy"
Now showing 1 - 20 of 22
Results Per Page
Sort Options
- Burkholderia phytofirmans strain PsJN effects on drought resistance, physiological responses and growth of switchgrassWang, Bingxue (Virginia Tech, 2015-02-09)To decrease dependency of fossil fuels and avoid direct competition with food crops, massive research efforts are investigating next-generation cellulose biofuel crops such as switchgrass (Panicum virgatum). A low-input, sustainable switchgrass production could be achieved by reducing traditional management practices though applying plant growth promoting rhizobacteria (PGPR), of which our understanding is still rather limited. To elucidate physiological mechanisms behind PGPR's beneficial effects, we inoculated switchgrass seedlings with Burkholderia phytofirmans strain PsJN. Two experiments were conducted to determine the initial and long-term responses of switchgrass to PsJN inoculation by tracking growth and leaf physiology. In a third experiments, we tested the effects of PsJN on growth and leaf-level physiology of switchgrass under a moderate pre-drought conditioning and a successive severe drought stress. PsJN inoculation increased biomass and promoted elongation of shoots within 17 days following inoculation. The enhanced root growth in PsJN inoculated plants lagged behind the shoot response, resulting in greater allocation to aboveground growth (p=0.0041). Lower specific root length (p=0.0158) and higher specific leaf weight (p=0.0029) were also observed in PsJN inoculated seedlings, indicating advanced development. Photosynthetic rates (Ps) were higher in PsJN inoculated seedlings after 17 days (54%, p=0.0016), which were related to higher stomatal conductance, greater water use efficiency, and lower non-stomatal limitation of Ps. These rapid changes in leaf physiology are at least partially responsible for switchgrass growth enhancement from PsJN treatment. The early growth enhancement in PsJN inoculated switchgrass linearly decreased with plant age. PsJN inoculation increased Ps of upper canopy leaves by 13.6% but reduced Ps of lower canopy leaves by 8.2%. Accelerated leaf senescence and early flowering were observed in PsJN-inoculated switchgrass, which might contribute to slightly lower aboveground biomass at final harvesting. Drought preconditioning increased Ps of PsJN-inoculated switchgrass during a later severe drought; whereas, control switchgrass only benefited from drought preconditioning when leaf water potential dropped below -1 MPa. This study verified early growth enhancement and accelerated development of switchgrass due to PsJN inoculation. Rapid improvement in leaf physiology is related to enhanced productivity. PsJN inoculation also improve drought tolerance of switchgrass.
- Characterization of activation tagged potato (Solanum tuberosum L.) mutantsAulakh, Sukhwinder Singh (Virginia Tech, 2012-09-14)Generation and characterization of activation tagged potato mutants could aid in functional genomic studies. Morphological and molecular studies were conducted to compare potato cv. Bintje, its two mutants, underperformer (up), and nikku generated using the activation tagging vector pSKI074, and nikku revertant plants. Mutant up exhibited a dwarf phenotype (plant height 42 cm vs. 73 cm in cv. Bintje), abundant axillary shoot growth (3.1 shoots/plant compared to 0.7 shoots/plant in cv. Bintje; in vitro plants), greater tuber yield, altered tuber traits and early senescence compared to wild-type Bintje under in vitro conditions. Under in vivo conditions, the dwarf and early senescence phenotypes of the mutant were consistent, but the tuber yield of up was less (250 g/plant compared to 610 g/plant in wild-type Bintje) and had fewer axillary shoots compared to wild-type (1.9 shoots/plant in up vs. 4.7 shoots/plant in Bintje). Mutant nikku plants exhibited an extremely dwarf phenotype (plant height 2 cm in nikku vs. 6 cm in Bintje), had small hyponastic leaves, were rootless, and infrequently produced small tubers when compared to cv. Bintje. The overall nikku phenotype was suggestive of a constitutive stress response, which was further supported by the higher expression levels of several stress-responsive genes in nikku. The nikku revertant plants exhibited near normal stem elongation, larger leaves and consistent rooting, and it was a case of partial reversion. Southern blot analyses indicated the presence of single T-DNA insertions on chromosome 10 in the up and on chromosome 12 in the nikku mutant. The reversion in the nikku plants was not associated with the loss of enhancer copies from the original nikku mutant. Reverse transcriptase PCR analyses indicated transcriptional activation/repression of several genes in the up and nikku mutants, suggesting pleiotropic effects. In revertant, the expression levels of several genes which were differentially regulated in the nikku mutant were similar to Bintje. The gene immediately flanking the right border of the T-DNA insertion, which encoded a novel BTB/POZ (Broad complex, Tramtrac, Bric a brac; also known as Pox virus and Zinc finger) domain-containing protein, was highly up-regulated in the up mutant. This protein domain plays an important role in several important developmental, transcriptional and regulatory pathways. The mRNA-seq analyses resulted in 1,632 genes that were differentially expressed between mutant up and Bintje and the total number of up-regulated genes (661) were less than the number of genes down-regulated (971 genes) in the up mutant. Further analyses indicated that a variety of biological processes including decreased cell division, cell cycle activity, and abiotic stress responses were modified in the up mutant. In the nikku mutant, two potato genes, encoding an Acyl-CoA N-acyltransferases (NAT) superfamily protein, and a predicted major facilitator superfamily protein (MFS) were identified and overexpression lines Bintje/35S::NAT1 and Bintje/35S::PMT1 were created for recapitulation of the nikku mutant phenotype. Methylated DNA-PCR between the nikku and the revertant indicated a change in methylation status of the 35S enhancers, suggesting that the nikku revertant phenotype may be associated with some epigenetic modification.
- Characterization of Polymorphic Microsatellites in Strawberry and Their Transferability to Other Genera in the Rosaceae FamilyArora, Vishal (Virginia Tech, 2006-02-09)We investigated the transferability of 20 Fragaria vesca microsatellite primer pairs to 13 Fragaria vesca accessions, six Fragaria species and ten commercially important species in Rosaceae. Genetic diversity studies were carried among 16 diploid Fragaria accessions using these polymorphic microsatellites. The average number of alleles amplified for a polymorphic locus was 4.7 with maximum being 8.0 and minimum being 3.0. Observed heterozygosity ranged from 0.00 to 0.84 with an average of 0.28. Expected heterozygosity ranged from 0.33 to 0.91 with an average of 0.76. Power of discrimination varied from 0.43 to 0.92 with an average of 0.78. Transferability of microsatellites to F. orientalis (4x) and F. Ã ananassa (8x) was high, i.e., 18 (90%) primers produced amplicons. Cross species amplification within Rosaceae using these primers showed limited transference. Four microsatellites showed amplification for different species in Rosaceae. Products generated by UDF-003 and UDF-018 primers were sequenced. Sequencing results for UDF-018 showed that three species, i.e., Pyrus calleryana, Prunus persica and Rubus idaeus contained the expected microsatellite whereas another four, i.e., Cotoneaster salicifolius, Rosa rugosa, Amelanchier arborea and Potentilla fruticosa had conserved regions resulting in generation of amplicons. For UDF 003, Spirea xbumalda and Prunus persica did not contain a microsatellite although there was some sequence similarity with Fragaria. Size homoplasy, i.e., alleles of identical size with different numbers of repeats within the SSR was observed among Fragaria and Rosaceae species for primer UDF-018, suggesting a need for caution when interpreting SSR variation from band migration in the absence of DNA sequences.
- Comparative genome analysis of Burkholderia phytofirmans PsJN reveals a wide spectrum of endophytic lifestyles based on interaction strategies with host plantsMitter, Birgit; Petric, Alexandra; Shin, Maria W.; Chain, Patrick S. G.; Hauberg-Lotte, Lena; Reinhold-Hurek, Barbara; Nowak, Jerzy; Sessitsch, Angela (Frontiers, 2013-04-30)Burkholdena phytohanans PsJN is a naturally occurring plant-associated bacterial endophyte that effectively colonizes a wide range of plants and stimulates their growth and vitality. Here we analyze whole genomes, of PsJN and of eight other endophytic bacteria. This study illustrates that a wide spectrum of endophytio life styles exists. Although we postulate the existence of typical endophytic traits, no unique gene cluster could be exclusively linked to the endophytic lifestyle. Furthermore, our study revealed a high genetic diversity among bacterial endophytes as reflected in their genotypic and phenotypic features. B. phytofirrnans PsJN is in many aspects outstanding among the selected endophytes. It has the biggest genome consisting of two chromosomes and one plasmid, well-equipped with genes for the degradation of complex organic compounds and detoxification, e.g., 24 glutathione-S-transferase (GST) genes. Furthermore, strain PsJN has a high number of cell surface signaling and secretion systems and harbors the 3-OH-PAME quorum sensing system that coordinates the switch of free-living to the symbiotic lifestyle in the plant-pathogen B. solanacearum. The ability of B. phytofirmans PsJN to successfully colonize such a wide variety of plant species might be based on its large genome harboring a broad range of physiological functions.
- Computer Vision Based Analysis of Broccoli for Application in a Selective Autonomous HarvesterRamirez, Rachael Angela (Virginia Tech, 2006-07-25)As technology advances in all areas of society and industry, the technology used to produce one of life's essentials - food - is also improving. The majority of agriculture production in developed countries has gone from family farms to industrial operations. With the advent of large-scale farming, the automation of basic farming operations has increasingly made practical and economic sense. Broccoli, which is still harvested by hand, is one of the most expensive crops to produce. Investing in sensing technology that can provide detailed information about the location, maturity and viability of broccoli heads has the potential to produce great commercial benefits. This technology is also a prerequisite for developing an autonomous harvester that could select and harvest mature heads of broccoli. This thesis details the work done to develop a computer vision algorithm that has the ability to locate the broccoli head within an image of an entire broccoli plant and to distinguish between mature and immature broccoli heads. Locating the head involves the use of a Hough transform to find the leaf stems and, once the stems are found, the location and extent of the broccoli head can be ascertained with the use of contrast texture analysis at the intersection of the stems. A co-occurrence matrix is then produced of the head and statistical texture analysis is performed to determine the maturity of the broccoli head. The conceptual design of a selective autonomous broccoli harvester, as well as suggestions for further research, is also presented.
- A Computer Vision Tool For Use in Horticultural ResearchThoreson, Marcus Alexander (Virginia Tech, 2017-02-13)With growing concerns about global food supply and environmental impacts of modern agriculture, we are seeing an increased demand for more horticultural research. While research into plant genetics has seen an increased throughput from recent technological advancements, plant phenotypic research throughput has lagged behind. Improvements in open-source image processing software and image capture hardware have created an opportunity for the development of more competitively-priced, faster data-acquisition tools. These tools could be used to collect measurements of plants' phenotype on a much larger scale without sacrificing data quality. This paper demonstrates the feasibility of creating such a tool. The resulting design utilized stereo vision and image processes in the OpenCV project to measure a representative collection of observable plant traits like leaflet length or plant height. After the stereo camera was assembled and calibrated, visual and stereo images of potato plant canopies and tubers(potatoes) were collected. By processing the visual data, the meaningful regions of the image (the canopy, the leaflets, and the tubers) were identified. The same regions in the stereo images were used to determine plant physical geometry, from which the desired plant measurements were extracted. Using this approach, the tool had an average accuracy of 0.15 inches with respect to distance measurements. Additionally, the tool detected vegetation, tubers, and leaves with average Dice indices of 0.98, 0.84, and 0.75 respectively. To compare the tool's utility to that of traditional implements, a study was conducted on a population of 27 potato plants belonging to 9 separate genotypes. Both newly developed and traditional measurement techniques were used to collect measurements of a variety of the plants' characteristics. A multiple linear regression of the plant characteristics on the plants' genetic data showed that the measurements collected by hand were generally better correlated with genetic characteristics than those collected using the developed tool; the average adjusted coefficient of determination for hand-measurements was 0.77, while that of the tool-measurements was 0.66. Though the aggregation of this platform's results is unsatisfactory, this work has demonstrated that such an alternative to traditional data-collection tools is certainly attainable.
- Development of intermonoploid somatic hybrids of potato and their molecular analysis based on polymorphism for retroelement Tst1Lightbourn, Gordon James (Virginia Tech, 2004-08-12)Inbred lines for hybrid crop production have been a mainstay of plant breeding. Biotechnological approaches to hasten the process are available including anther culture to halve the genome and protoplast fusion to create hybrids between incompatible partners. We applied these techniques to potato to evaluate their potential for breeding highly heterozygous, cross-pollinating species. Four families of monoploids (2n=1x=12), developed from diploid hybrids with diverse genomic constitutions but heavily favoring Solanum phureja, a primitive cultivated potato, were used in electrofusion experiments to create intermonoploid somatic hybrids (SH). The "monoploid sieve" results in the survival of only those gametes free of lethal and deleterious genes but generates sterile sporophytes, necessitating protoplast fusion for SH development. From six intermonoploid electrofusion combinations, 276 plants were regenerated over 6-9 months. Fusion conditions were optimized. Ploidy was determined by flow-cytometry and SH confirmed by microsatellite analysis. Field evaluations over three years revealed that intermonoploid SH were inferior to cultivars. Dihaploids derived by anther culture of a tetraploid intermonoploid SH were reduced in vigor with an increase in homozygosity, while 2x X 2x sexually derived populations had better yield than the SH, suggesting that producing SH introduced or eliminated factors required for productivity. Molecular analysis of the SH was conducted to examine genomic stability through protoplast isolation and plant regeneration. Sequence specific amplified polymorphism (S-SAP) represents a hybrid system incorporating amplified fragment length polymorphism (AFLP) technology in conjunction with the use of a defined genomic sequence, e.g., retrotransposon display (RD) when the defined sequence is anchored into a consensus sequence of a retrotransposon such as the long terminal repeat (LTR) sequence of Tst1. Parental monoploids, SH and various Solanaceae were evaluated by RD. Fluorescently-labeled retrotransposon-based primers were used in the ALFexpress automated fragment analyzer system. Eleven probes from RD were created for Southern blot analysis and used to verify taxonomic relationships between selected Solanaceae. Blots of intermonoploid somatic hybrids confirmed hybridity and occasional loss of genomic fragments. No activation or replication of retrotransposons was detected. Sequencing of inter-retrotransposon amplified polymorphism (IRAP) and S-SAP fragments revealed that all fragments had the expected Tst1 retroelement and/or the AFLP adaptor sequence. BLAST analysis identified 4 of the 17 fragments sequenced as part of the chloroplast genome, a tobacco anther-specific gene, repetitive DNA, and the phytochrome F gene.
- Function and Regulation of Xylem Cysteine Protease 1 and Xylem Cysteine Protease 2 in ArabidopsisIsmail, Ihab (Virginia Tech, 2003-04-04)A functional water-conducting system, the tracheary elements of the xylem, is required to sustain plant growth and development. Tracheary element formation is dependent on many biological processes terminated by programmed cell death and cellular autolysis. The final two processes are probably dependent on the activity of hydrolytic enzymes such as XCP1 and XCP2 known to be expressed in tracheary elements during these final two processes. Thus, the transcriptional regulation of XCP1 and the function of XCP2 were investigated. Qualitative and quantitative assessments of GUS activity as directed by various fragments of the XCP1 promoter showed that a 237-bp internal region was able to drive GUS expression in a tracheary element-specific manner in Arabidopsis. A 25-bp deletion at the 3' end of this region abolished GUS expression. The 237-bp region served as bait in a yeast one-hybrid analysis. Screening of yeast colonies retrieved 109 putative positive interactions, which included a potential transcriptional regulator, indole acetic acid-induced protein 8 (IAA8). An auxin responsive element that potentially binds auxin responsive transcription factors was found within the 25-bp deletion. Cis-elements were predicted by Genomatix and Athamap computer programs. The cis-elements form pyrimidine and gibberellic acid responsive elements that can potentially bind Dof and Myb transcription factors, respectively. In an independent effort, attempts to develop a mapping population to isolate upstream regulators of XCP1 expression did not succeed. Functionally, tracheary element-specific expression of XCP2 in Arabidopsis suggested a specialized role for XCP2 in final phases of tracheary element differentiation. The function of XCP2 was assessed using T-DNA insertional mutants, post-transcriptional gene silencing, and through tracheary element-specific expression of the cysteine protease inhibitor, soyacystatin N in Arabidopsis. Our findings revealed that the absence of XCP2 expression due to T-DNA insertional mutagenesis did not affect plant growth and development in the laboratory. Soyacystatin N was an effective in vitro inhibitor of cysteine proteases. Plants expressing 35S-driven cytosolic form of soyacystatin exhibited stunting and reduced apical dominance. Plants expressing pXCP1-driven cytosolic soyacystatin did not differ from wild type plants. Additionally, transgenic plants expressing pXCP1- and 35S-directed XCP2-double-stranded RNA for the silencing of XCP2 showed no unusual phenotypes compared to their wild type counterparts
- Gametophytic Selection for Thermotolerance in PhalaenopsisBlischak, Leslie Anne (Virginia Tech, 2005-07-06)Gametophytic selection was examined as a breeding tool in developing Phalaenopsis hybrids that are more cool or warm temperature tolerant. Two hybrid Phalaenopsis, P. (Taisoco Windian à Sogo Sogo Yukidian) by P. hybrid unknown, were reciprocally cross-pollinated and exposed to 14°C/9°C for 7 days as a cold pollination treatment. Plants were pollinated again and exposed to 30°C/25°C for 3 days for the warm pollination treatment. Each cultivar was placed in either of two growth chambers during the pollination treatments and exposed to the selected temperatures, an 11-h photoperiod with an irradiance of 180 Mmol·m-2·s-1 and a relative humidity of 70%. The plants were returned to the greenhouse after pollination and the green capsules were collected after 150 days. Seeds obtained from these treatments were surface-sterilized and equal volumes were placed on Phytamax® medium. Evaluation of protocorm development was done after 73 days on a thermogradient table ranging from 10 to 30ºC. For the first family for which reciprocal crosses were available, the number of protocorms per plate ranged from 0 in the coldest treatments to 290 at 28°C. For cold pollinated seeds, protocorm development was optimum at 22 and 28°C (means of 290 and 250 protocorms per plate, respectively) whereas the greatest protocorm development for warm pollinated seeds occurred at 20°C (103 protocorms per plate). Of the 1471 total protocorms obtained 1095 were from cold pollinations, whereas 376 were from the warm pollinations. Protocorms were evaluated for leaf and root formation 125 days after initial plating. Transfer to warm or cold incubators occurred as protocorms developed leaves and roots. Seedlings were finally transferred to dried sphagnum and placed in growth chambers set to original pollination temperatures. One year after initial plating seedlings were evaluated on the following criteria: wet weight, number of leaves, leaf area, number of roots, and root length. The pollination treatment significantly affected the number of roots per seedling whereas germination temperature during germination significantly affected the weight (g). Weight of the seedlings, number of roots and the average root length were significantly affected by the interaction between pollination treatment and germination temperature. The weight, number of leaves, and average root length were significantly affected by the interaction between pollination treatment and incubator/growth chamber. These differences indicated that seedlings derived from warm pollination were more vigorous under warm growing conditions and those derived from cold pollination were more vigorous under cold growing conditions. The significance of the interaction between pollination treatment and incubator/growth chamber indicates that gametophytic selection for thermotolerance in Phalaenopsis can be successfully used as a plant breeding tool. Additional replication is required to confirm the greater germinability of seed derived from pollination occurring over a greater range of temperatures.
- Growth promotion and colonization of switchgrass (Panicum virgatum) cv. Alamo by bacterial endophyte Burkholderia phytofirmans strain PsJNKim, Seonhwa; Lowman, James Scott; Hou, Guichuan; Nowak, Jerzy; Flinn, Barry; Mei, Chuansheng (2012-05-30)Background Switchgrass is one of the most promising bioenergy crop candidates for the US. It gives relatively high biomass yield and can grow on marginal lands. However, its yields vary from year to year and from location to location. Thus it is imperative to develop a low input and sustainable switchgrass feedstock production system. One of the most feasible ways to increase biomass yields is to harness benefits of microbial endophytes. Results We demonstrate that one of the most studied plant growth promoting bacterial endophytes, Burkholderia phytofirmans strain PsJN, is able to colonize and significantly promote growth of switchgrass cv. Alamo under in vitro, growth chamber, and greenhouse conditions. In several in vitro experiments, the average fresh weight of PsJN-inoculated plants was approximately 50% higher than non-inoculated plants. When one-month-old seedlings were grown in a growth chamber for 30 days, the PsJN-inoculated Alamo plants had significantly higher shoot and root biomass compared to controls. Biomass yield (dry weight) averaged from five experiments was 54.1% higher in the inoculated treatment compared to non-inoculated control. Similar results were obtained in greenhouse experiments with transplants grown in 4-gallon pots for two months. The inoculated plants exhibited more early tillers and persistent growth vigor with 48.6% higher biomass than controls. We also found that PsJN could significantly promote growth of switchgrass cv. Alamo under sub-optimal conditions. However, PsJN-mediated growth promotion in switchgrass is genotype specific. Conclusions Our results show B. phytofirmans strain PsJN significantly promotes growth of switchgrass cv. Alamo under different conditions, especially in the early growth stages leading to enhanced production of tillers. This phenomenon may benefit switchgrass establishment in the first year. Moreover, PsJN significantly stimulated growth of switchgrass cv. Alamo under sub-optimal conditions, indicating that the use of the beneficial bacterial endophytes may boost switchgrass growth on marginal lands and significantly contribute to the development of a low input and sustainable feedstock production system.
- Integration of High Residue/No-till and Farmscaping Systems in Organic Production of BroccoliBenson, Gordon Brinkley (Virginia Tech, 2006-05-01)High-biomass cover cropping enhances marketable yields in organic production of vegetables, linked to the improvement of soil quality and weed control. Although, during transition from chemical to organic cover-cropping production, especially with no-till systems, reduction of nitrogen availability to the main crop and increase in weed and pest pressure may occur. In 2004-2005, summer and fall broccoli (Brassica oleraceae L. Botrytis Group) crops were grown in twin rows on permanent (controlled traffic) raised beds (185-cm wide). Before broccoli transplanting, high-biomass cover crops were grown in specific bed areas. Legumes (Vicia villosa, Glycine max L., and Crotolaria juncea L.) on bed tops (grow zones) and grass species (Secale Cereale L., Setaria italica L., and Sorghum bicolor X S. bicolor var. Sudanese) in the alleyways (bed shoulders and bottoms). Experimental treatments were tillage (conventional, CT; and no-tillage, NT), farmscaping (with and without), and nitrogen sidedressing (with and without, applied 3 weeks after transplanting as a mixture of sodium nitrate - 22 kg N ha-1 - and feathermeal - 44 kg N ha-1). Weeds were managed by mechanical cultivation in CT and a spot weeding by hand in NT treatments. High numbers of beneficial insects (Cotesia glomerata, Cotesia orobenae and Diadegma insulare) kept the primary insect pest population (Pieris rapae, Evergestis rimosalis (Guenee), and Plutella xylostella) at a pest to predator ratio below 4:1. Although the excellent insect pest control was attributed to the farmscape plantings, pest level and crop yields were not significantly affected by farmscaping (likely due to the close proximity of the farmscaped plots (10-50m from non-farmscaped)). Broccoli yield averaged 62% higher in fall than summer (12.1 vs. 7.5 t ha-1) likely due to cool weather conditions during broccoli head development (October), increasing head size, uniformity, and marketability. In 2004, broccoli yield in CT plots was either equal or slightly higher than NT (9.5 vs. 9.0 t ha-1). However, in 2005, broccoli yield in CT plots was significantly higher in both spring (8.8 vs. 6.8 t ha-1; p = 0.0258) and fall crops (13.5 vs. 12.3 t ha-1 with p = 0.0484). Nitrogen sidedressing improved yield in all plots (9.8. vs. 12.7 t ha-1) and particularly in NT (8.6 vs. 12.1 t ha-1), indicating that availability and/or synchrony of nitrogen was a limiting factor. Incorporating high-N legume residues in the grow zones resulted in a lesser N response in CT.
- Micro-Level Management of Agricultural Inputs: Emerging ApproachesWeekley, Jonathan; Gabbard, Joseph L.; Nowak, Jerzy (MDPI, 2012-12-18)Through the development of superior plant varieties that benefit from high agrochemical inputs and irrigation, the agricultural Green Revolution has doubled crop yields, yet introduced unintended impacts on environment. An expected 50% growth in world population during the 21st century demands novel integration of advanced technologies and low-input production systems based on soil and plant biology, targeting precision delivery of inputs synchronized with growth stages of crop plants. Further, successful systems will integrate subsurface water, air and nutrient delivery, real-time soil parameter data and computer-based decision-making to mitigate plant stress and actively manipulate microbial rhizosphere communities that stimulate productivity. Such an approach will ensure food security and mitigate impacts of climate change.
- Multispectral Imaging Techniques for Monitoring Vegetative Growth and HealthWeekley, Jonathan Gardner (Virginia Tech, 2007-12-04)Electromagnetic radiation reflectance increases dramatically around 700 nm for vegetation. This increase in reflectance is known as the vegetation red edge. The NDVI (Normalized Difference Vegetation index) is an imaging technique for quantifying red edge contrast for the identification of vegetation. This imaging technique relies on reflectance values for radiation with wavelength equal to 680 nm and 830 nm. The imaging systems required to obtain this precise reflectance data are commonly space-based; limiting the use of this technique due to satellite availability and cost. This thesis presents a robust and inexpensive new terrestrial-based method for identifying the vegetation red edge. This new technique does not rely on precise wavelengths or narrow wavelength bands and instead applies the NDVI to the visible and NIR (near infrared) spectrums in toto. The measurement of vegetation fluorescence has also been explored, as it is indirectly related to the efficiency of photochemistry and heat dissipation and provides a relative method for determining vegetation health. The imaging methods presented in this thesis represent a unique solution for the real time monitoring of vegetation growth and senesces and the determination of qualitative vegetation health. A single, inexpensive system capable of field and greenhouse deployment has been developed. This system allows for the early detection of variations in plant growth and status, which will aid production of high quality horticultural crops.
- Promoter Deletion Analysis of Xylem Cysteine Protease 2 (XCP2) in Arabidopsis thalianaPetzold, Herman Earl III (Virginia Tech, 2007-05-02)The process of xylem tracheary element differentiation involves the coordination of vascular cambium activity, cell fate determination, cell expansion/elongation, secondary wall synthesis, programmed cell death, and cellular autolysis. The end result of tracheary element differentiation is a cellular corpse lacking a protoplast and consisting of a thickened cell wall composed mostly of lignin and cellulose. Little is known about the genetic mechanisms regulating the process of tracheary element differentiation. XCP2 expression localizes to tracheary elements according to two independent methods of analysis: promoter reporter experiments and immunogold localization by electron microscopy. XCP2 may be involved in catalyzing the degeneration of the protoplast during the final autolytic stages of tracheary element differentiation. To this date XCP2 function has not been directly demonstrated. In principle, any tracheary element-specific markers can be linked to upstream regulatory genes with roles in tracheary element differentiation. To develop the XCP2 promoter as a tool for identification of transacting factors, a promoter deletion analysis was carried out. Utilizing information from 5â and 3â deletion constructs, a 70-bp region upstream of the XCP2 translational start site is both necessary and sufficient for TE-specific expression of the UidA reporter gene. Mutational analysis of the ACTTTA element at position -113-bp strongly suggests it is a cis element required for XCP2 expression. In silico analysis of an 18-bp promoter region located within 200-bp of the translation start site and including the ACTTTA element revealed high indentity shared between xylem-specific XCP2 homologs from Zinnia elegans, Populus trichocarpa, and XCP1 from Arabidopsis thaliana.
- Resident Advisors' Attitudes and Beliefs About the Process of Identifying and Reporting Threatening BehaviorsCasten, Jill Nicole (Virginia Tech, 2011-03-29)Threatening behavior is a cause for concern on college campuses. Even though solutions presented cannot completely prevent crime and violence, steps must be taken towards continuous improvement of violence prevention efforts. The purpose of this case study was to examine resident advisors' attitudes and beliefs about the process of identifying and choosing to report threatening behaviors. Thirteen participants from Residence Life at a major Land Grant University served as the case study group. A qualitative approach guided the inquiry of the study and was represented through observations, document analysis, and participant interviews. Analysis of the research questions supported the study's findings. Overall, students' exposure to violence prevention on campus is often through educational programs and resources, while Resident Advisors are also instrumental resources in sharing information with students. They suggest making more efforts in sharing violence prevention education with students. Because a wide range of issues face today's college students, they seek assistance from Resident Advisors, as well as peers, friends, and family. Building community through the residence halls provides a proactive approach in efforts to develop and provide students opportunities for growth, responsibility, and accountability to their communities. Finally, despite reporting systems in place and resources available, barriers still remain for students. The inconsistency between the views and perceptions of violence prevention education and barriers to reporting between Resident Advisors and administration in Residence Life indicates need for further engagement between students and Institutions of Higher Education based upon policies, educational efforts, and reporting structures. Institutions have an obligation to engage students in the prevention of violence through sustained community-building measures and working with targeted peer groups, such as Resident Advisors. Conclusions from this study were explained through the individual, information, and social background factors of The Reasoned Action Approach and guided the recommendations for practice and further research. Addressing the disconnect between the administration and Resident Advisors' perceptions is crucial in reducing barriers to reporting. A need remains for further engagement with peers, friends, family, RAs, and other influential groups to help shape students' understanding, awareness, and continued involvement in the identification and reporting of threatening behaviors.
- Seeds : Newsletter for Alumni of the Department of Horticulture at Virginia Tech : Fall 2006Nowak, Jerzy; Shelton, Joyce (Virginia Tech. Department of Horticulture, 2006)I hope all our Alumni had a successful year and are looking forward to receiving good news from the Home Department. I am very pleased to share with you some of our successes. The Peggy Lee Hahn Garden Pavilion was completed in June and formally dedicated on June 15 (http://www.vtnews.vt.edu/story.php? relyear=2006&itemno=344). This beautiful structure attracts university, town, New River communities, alumni and professional organizations throughout the state. Our ambitious plan is to make it a center for the expansion of consumer horticulture programs. On September 22, the Colleges of Agriculture and Life Sciences and Natural Resources celebrated the opening and dedication of Latham Hall, a new 84,000-square-foot multidisciplinary research facility (http://www.vtnews.vt.edu/ story.php?relyear=2006&itemno=515). I am pleased to let you know that our Department secured five laboratories in this new building, accommodating research and graduate programs of six faculty. The labs of Drs. Eric Beers, Jim Tokuhisa, and Richard Veilleux have already been established and searches for three additional faculty (Specialty Crops Breeder for Bioenergy/Bioproducts, Ornamental Plant Breeder, and Rhizosphere Biologist) are in progress. We would like to renovate the vacated space in Saunders Hall and convert it into a modern undergraduate teaching and research facility.
- Seeds : Newsletter for Alumni of the Department of Horticulture at Virginia Tech : Fall 2007Nowak, Jerzy; Shelton, Joyce (Virginia Tech. Department of Horticulture, 2007)The University and the Department went through a traumatic experience on April 16. The tragedy has unified Virginia Tech's community probably stronger than ever before. Several memorial projects are now under development that include participation of students, victim families, faculty, staff, alumni, and others across the US and beyond. One project is a Hokie Spirit Garden Trail that will integrate green spaces on campus, including proposed development of new theme gardens, and link the Trail to the Huckleberry Trail. Both walking and biking trails are considered. The project is a partnership between the campus community and the green industry. The Horticulture Department plays a significant role in this undertaking as well as in the development of a Jocelyne Couture-Nowak Memorial Garden as a part of the Terrace Garden currently under construction at the Peggy Lee Hahn Garden. Here I would like to take the opportunity to thank the alumni for the support you gave me and my family after my wife’s tragic death. Your sympathy and kindness helped carry us through these darkest hours and have been a source of our strength. Here is the url of the article on our family which has been recently published in the Chronicle of Higher Education: http://chronicle.com/free/v54/i04/ 04a00101.htm.
- Seeds: A Publication for Alumni of the Department of Horticulture at Virginia Tech : Fall 2005Nowak, Jerzy; Shelton, Joyce (Virginia Tech, 2005)
- Total fatty acid production in golden alga Prymnesium parvum a potential bio-diesel feedstockCulver, Bruce (Virginia Tech, 2009-05)Developing renewable and sustainable feedstocks for biodiesel production is one strategy for reducing our dependence on petroleum-based fuel. Microalgae are attractive feedstocks for biodiesel due to their high lipid content and fast growth rate. In addition algae, can be cultured on marginal lands using saline water and thus avoiding competition with arable land. This study investigates the total fatty acid content in Prymnesium parvum, which has the potential as a feedstock for biodiesel production. Prymnesium parvum was cultured in artificial sea water with the salinity adjusted to 14 - 15 (psu) and a pH ranging from 7.8 - 8.5. The culture temperature was 25.7°C under florescent light with a 16:8 (L/D) cycle. Total fatty acids produced had a mean of 11.679 % ± 0.68671 (DW). These percentages were below the 22-38% previously reported by E. W. Becker 1994. Total algal biomass produced had a mean yield of 0.34mg ± 0.07204 (DW) / L. In comparison to other algal species used as feedstocks, P. parvum produced significantly less fatty acids and biomass. Prymnesium parvum also exhibited signs of self toxicity with frequent crash cycles. The self toxicity would have contributed to its inability to produce both lipids and dry biomass. Further studies for increasing fatty acid production in P. parvum may require using a continuous bioreactor system, controlling environmental conditions to relieve self-toxicity and adjusting nutrient concentrations. Exposure to different light/dark regiments along with increasing CO2 concentrations should be explored to possibly increase yields.
- Towards Identifying Cis and Trans Regulators of Expression of Xylem Cysteine Protease 1 (XCP1) in ArabidopsisStroud, William Jefferson (Virginia Tech, 2009-05-06)Secondary xylem, commonly known as wood, is a valuable commercial commodity. Among the major components of wood are the elongated, thick-walled water-conducting cells known as tracheary elements. Understanding tracheary element differentiation and maturation is of scientific and commercial importance as it may lead to broad understanding of cellular differentiation processes as well as ways to increase both the quality and quantity of wood produced by economically important tree species. One way to begin to understand the regulation of tracheary element differentiation is to identify elements that control expression of genes associated with tracheary elements. In Arabidopsis thaliana, Xylem Cysteine Protease 1 (XCP1) is specifically expressed in tracheary elements where it catalyzes microautolysis. Thus XCP1 can serve as a useful model for identifying factors that regulate tracheary element-specific gene expression. A deletion analysis of the XCP1 promoter was conducted to identify promoter elements that are necessary and sufficient for tracheary element-restricted gene expression. Two regions required for tracheary element-specific gene expression were identified. One of these was assembled as a multimeric bait construct and used in yeast one-hybrid assays to identify candidate transcription factors that bind to the XCP1 promoter region. Subsequently, a southwestern blot analysis was used to identify transcription factors displaying specific binding to a previously reported cis-element, CTTCAAAGCCA, found in the XCP1 promoter and other tracheary element-associated genes from multiple species.