Switchgrass (Panicum virgatum L.) is a candidate bioenergy crop. Somatic embryogenic (SE) calli are used for genetic transformation in switchgrass. A superior switchgrass line, HR8, was developed using recurrent tissue culture selection from cv. Alamo. HR8 SE calli were genetically transformable using Agrobacterium at an efficiency of ~12%. We used HR8 somatic embryogenic calli for genetic improvement of switchgrass. The lignin content of feedstock has been proposed as one key trait impacting biofuel production. 4-Coumarate: Coenzyme A ligase (4CL) is one of the key enzymes involved in the monolignol biosynthetic pathway. Two homologous 4CL genes, Pv4CL1 and Pv4CL2, were identified in switchgrass. Gene expression patterns and enzymatic activity assays suggested that Pv4CL1 is involved in monolignol biosynthesis. Stable transgenic plants were obtained with Pv4CL1 down-regulated. RNA interference of Pv4CL1 reduced extractable 4CL activity by 80%, leading to a reduction in lignin content with decreased guaiacyl unit composition. The transgenic plants had uncompromised biomass yield. After dilute acid pretreatment, the low lignin transgenic biomass had significantly increased cellulose hydrolysis (saccharification) efficiency for biofuel production. Erect leaf is a desirable trait to adjust the overall plant architecture to perceive more solar energy and thereby to increase the plant biomass production in a field population. We overexpressed an Arabidopsis NAC transcriptional factor gene, LONG VEGETATIVE PHASE ONE (AtLOV1), in switchgrass. Surprisingly, AtLOV1 induced smaller leaf angle by changing morphologies of epidermal cells in the leaf collar region, affecting lignin content and monolignol composition, and also causing delayed flowering time in switchgrass. Global gene-expression analysis of AtLOV1 transgenic plants demonstrated an array of genes has altered expressions. Potential downstream genes involved in the pleiotropic phenotypic traits of the transgenic plants are discussed. / Ph. D.

<p>In recent years, hyperspectral imaging technologies have been broadly applied to evaluate complex plant physiological features such as leaf moisture content, nutrient level and disease stress. A  critical  component  of  this  technique  is  white  referencing  used  to  remove  the  effect  of  non-uniform  lighting  intensity  in  different  wavelengths  on  raw  hyperspectral  images. Based  on  the  literature,  the leaf  geometry (e.g.,  tilt  angles)  and its interaction  with  the  illumination  severely impact  the  plant  reflectance  spectra  and vegetation  indices  such  as  the  normalized  difference  vegetation index (NDVI).  This thesis is  aimed to address the issues caused by the tilt angles across the leaf surface. To achieve this, two methods based on the fusion of the hyperspectral images and 3D  point  clouds  were  proposed.  The  first  method  was  to  build  a  3D  white  reference  library  in  which a point with almost the same tilt angle, height and position with the pixel on the plant leaf can be found, and then the white reference spectrum at that point can be used to calibrate the raw spectrum of the leaf pixel. The second method was to observe and summarize how the plant spectra and NDVI values changed with the leaf angles. Using the changing trends, the original NDVI and spectra  of  leaf  pixels  at  different  angles  can  be  calibrate  to  a  same  standard  as  if  the  leaf  was  imaged  at  a  flat  and  horizontal  surface.  The  approach  was  called  3D  calibration.  The  results  showed  that  the NDVI  values significantly  changed  with  leaf  angles  and  the  changing  trends differed  between  the  corn  and  soybean  species.  To evaluate the  performance  of  3D  calibration, 180 soybean plants with different genotypes, nitrogen (N), phosphorus (P) and water treatments were  grown  in  the  greenhouse. Each  plant  was  imaged  in three systems:  the high-throughput greenhouse hyperspectral imaging system, the indoor desktop imaging system with a visible-near infrared  (VINIR)  hyperspectral camera and  an  Intel  RealSense  depth  camera  and  the handheld device hyperspectral imaging system. In the greenhouse system, the whole canopy was captured. In the indoor desktop system, the partial canopy was captured because of the space limitation and the  top-matured  leaf  (the  middle  leaf  of  the  uppermost  matured  trifoliate)  was  focused.  The proposed  3D  calibration  was  applied  on  the  top-matured  leaf  to  remove  angle  impacts.  In  the  handheld device system, the flat top-matured leaf was captured. After done with imaging work, the plants were harvested to collect the ground truth data such as relative water content (RWC), N content and P content. Combined with the ground truth data, the NDVI values from three systems were  used  to  discriminate  different  genotypes  and  biochemical treatments,  whereas,  the  spectra from three systems were used to build partial least squares regression (PLSR) models for N, P and RWC. The results showed that the averaged tilt angles of top-matured leaves were impacted by different treatments. For instance, the low-nitrogen (LN) plants showed significantly higher leaf angles than high-nitrogen (HN) plants; the leaf angles on water-stressed (WS) plants were higher than those on well-watered (WW) plants. The leaf angles carried some signals that influenced not only the NDVI discrimination but also the PLSR modelling results. The signals were lost after 3D calibration.  For  the  top-matured  leaves,  the  discrimination  and  modelling  results  after  3D  calibration in the indoor desktop system were close to those from the flat leaves in the handheld device  system.  The  proposed  3D  calibration  approach  has  a  potential  to  eliminate  leaf  angle  impacts.</p>

Submitted by (edna.saturno@ufrpe.br) on 2017-03-23T12:40:14Z No. of bitstreams: 1 Bruno Leal Viana.pdf: 1623265 bytes, checksum: 5707cff554bc2506977fb551f2f6b45e (MD5) / Made available in DSpace on 2017-03-23T12:40:14Z (GMT). No. of bitstreams: 1 Bruno Leal Viana.pdf: 1623265 bytes, checksum: 5707cff554bc2506977fb551f2f6b45e (MD5) Previous issue date: 2009-07-27 / The research was conducted at the Experimental Station of the Agronomic Institute of Pernambuco (IPA) located in Itambé, dry rainforest region of Pernambuco. The objective was to estimate the coefficients of repeatability, of determination (R2) and the number of measures needed to predict the real value [using principal component analysis (PCA)] to variable morphophysiological and productive, and the responses dwarf elephant grass pasture under grazing sheep. The experiment was carried out in a randomized block design with five Pennisetum purpureum clones were evaluated: Taiwan A-146 2.27, Taiwan A-146 2.37, Taiwan A-146 2114, Merker Mexico MX 6.31 and Mott. Evaluations were performed during five grazing cycles, from September 2008 to April 2009. Twenty five sheep (42 kg average liveweight) were used in a mobgrazing protocol, leaving the paddock at 30 cm of residual stubble height. The clones were managed under rotational stocking, with 32 days of resting period and three days of grazing period for cycles 1, 2, and 5, and 64 days of resting period and two days ofgrazing period for cycles 3 and 4. Response variables analyzed at pre- and post-grazing included leaf area index, light interception, leaf angle, average sward height, green portion of herbage mass (DM basis). In addition, we estimated net herbage accumulation (ALF), in kg DM ha-1 cycle-1, and net herbage accumulation rate (TALF), in kg DM ha-1 day-1. Data were analyzed by univariate or multivariate analysis of repeated measures over time in the SAS. Means were compared by Tukey test at 5% probability. According to the PCA, for the variables LAI and LI in cycles of shorter period of regrowth, and height of plants in the cycles of higher period of rest is needed only one measurement (R2=0.80), classifying them among the characteristics evaluatedas the easiest prediction in elephant grass clones. Assessments for NHA and NHAR size of dwarf elephant grass, under grazing, need long experimental periods. / Esta pesquisa foi realizada na Estação Experimental do Instituto Agronômico de Pernambuco (IPA), no município de Itambé, Zona da Mata Seca de Pernambuco, com o objetivo de estimar os coeficientes de repetibilidade, de determinação (R2) e o número mínimo de medições necessárias para predizer o valor real [pela análise dos componentes principais (ACP)] para variáveis morfofisiológicas e produtivas, bem como as respostas dos pastos de capim elefante de porte baixo sob o pastejo de ovinos. O experimento foi conduzido no delineamento em blocos casualizados, com cinco clones de Pennisetum purpureum (Taiwan A-146 2.27, Taiwan A-146 2.37, Taiwan-146 2.114, Merker México MX 6.31, e Mott). As avaliações foram realizadas em cinco ciclos de pastejo, compreendidos entre os meses de setembro de 2008 a abril de 2009. Foram utilizados 25 ovinos, com peso médio de 42 kg, como animais pastejadores em “mobgrazing”, buscando-se a altura de 30 cm de resíduo pós-pastejo. Os clones foram manejados sob lotação intermitente, sendo 32 dias de descanso e três de pastejo para os ciclos 1, 2 e 5 e 64 dias de descanso e dois de pastejo para os ciclos 3 e 4. As variáveisanalisadas foram índice de área foliar, interceptação luminosa, ângulos foliares médios realizados, alturas médias do dossel e massa de forragem seca verde (MSV), no pré e pós pastejo. Foram estimados ainda o acúmulo líquido de forragem (ALF), em kg MS ha-1 ciclo-1 e a taxa de acúmulo líquido de forragem (TALF), em kg MS ha-1 dia-1. Os dados foram analisados por meio de análise univariada ou multivariada de medidas repetidas no tempo no SAS. Para comparação das médias foi utilizado o teste de Tukey a 5% de probabilidade. De acordo com a ACP, para as variáveis IAF e IL nos ciclos de menor período de rebrota, e altura média de plantas nos ciclos de maior período de descanso é necessário apenas uma medição (R2=0,80), classificando-as entre às características avaliadas como as de mais fácil predição em clones de capim elefante. Avaliações de ALF e de TALF para capim elefante de porte baixo, sob pastejo, necessitam de períodos experimentais mais longos.

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