Browsing by Author "Brunner, Amy M."

Now showing 1 - 19 of 19
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    Architecture and Evolution of Xylem-related Gene Coexpression Networks in Poplars
    Suren, Haktan (Virginia Tech, 2013-02-14)
    With the advent of sequencing technologies, a variety of methods have been available day by day. Each of these methods have helped scientists to for a deeper understanding of the biological function and evolutionary constraints on the relevant genes, which can be gained through the use of modern computational approaches. Numerous approaches have being developed to advance these goals, and interaction network mapping is one of them. This method has been employed to study a variety of organisms to illustrate shared (conserved) or individual (unique) properties, and is mainly based on identifying and visualizing modules of co-expressed genes. As being a very strong candidate for such tools, co-expression gene network was used in this study to indentify the genes in wood formation of Populus trichocarpa with the help of the other novel bioinformatics tools such as Gene Ontology and Cytoscape. In order to booster the accuracy of the findings, we have combined it with an evolutionary approach, synonymous and non-synonymous ratio (dN/dS) of the proteins to show the selective patterns of the genes in a comparative fashion between woody and non-woody plants. This thesis is proposed to help plant scientists to gain insights into the genes that are involved in wood formation. By taking advantage of the computational studies have been done on this paper, one can validate the experiments along with reducing the cumbersomeness of the lab trials on the topic of wood formation in plants
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    Functional analysis of novel protein-protein interactions involving ROP GTPases in Arabidopsis thaliana and Populus trichocarpa
    Jia, Xiaoyan (Virginia Tech, 2013-09-02)
    We are using the yeast two-hybrid (Y2H) system to identify novel protein-protein interactions (PPI) relevant to wood formation. Bait proteins for Y2H binary assays and screening against a xylem cDNA prey library were selected from approximately 400 Populus trichocarpa genes that are at least 8-fold more highly expressed in differentiating secondary xylem versus phloem-cambium, and designated here as poplar biomass (PB) genes. Here we report some of the interactions involving selected PB proteins and efforts to characterize their functions in Populus and Arabidopsis. Members of the ROP GTPase family, PB15 in poplar and ROP11 in Arabidopsis, interact with the domain of unknown function (DUF) 620 (DUF620) proteins (e.g., PB129 in poplar). Ectopic co-expression of PB15 and PB129 in Arabidopsis caused outgrowths at the base of flower pedicels and altered leaf morphology. Interestingly, the co-expression phenotype could not be observed in transgenic plants that are only expressing either one of the interacting partners separately. Transgenics altered in expression of PB15 and/or PB129 were prepared in Populus and characterization of transgenic trees will be performed in greenhouse and field. In addition to DUF620 family proteins, ROP11 also interacts with the COP9 subunit CSN5A in Arabidopsis. We confirmed the interaction of ROP11 and CSN5A in Y2H and employed available mutants for ROP11 and CSN5A in Arabidopsis to genetically characterize this interaction. Surprisingly, loss of ROP11 was found to rescue the csn5a-2 pleiotropic phenotype. Ectopic expression of a ROP11 dominant negative mutant in the csn5a-2 background also complemented the stunted growth phenotype. Transcript analysis and gel blot assays showed that CSN5A transcript levels remained unchanged in all rescue lines, whereas CSN5A protein levels increased relative to WT. Taken together, we concluded that ROP11 negatively regulate CSN5A protein level in plant by some as yet unknown mechanism.
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    Functional characterization of NAC-domain transcription factors implicated in control of vascular cell differentiation in Arabidopsis and Populus
    Grant, Emily H. (Virginia Tech, 2008-12-10)
    Wood has a wide variety of uses and is arguably the most important renewable raw material. The composition of xylem cell types in wood determines the utility of different types of wood for distinct commercial applications. Using expression profiling and phylogenetic analysis, we identified many xylem-associated regulatory genes that may control the differentiation of cells involved in wood formation in Arabidopsis and poplar. Prominent among these are NAC-domain transcription factors (NACs). In addition to their roles as regulators of xylem differentiation, NACs are regulators of meristem development, organ elongation and separation. We studied a subset of Populus and Arabidopsis NACs with putative involvement in xylem cell expansion and elongation (XND1/ANAC104, PopNAC118, PopNAC122, PopNAC128, PopNAC129), and secondary cell wall synthesis (ANAC073, PopNAC105, PopNAC154, PopNAC156, PopNAC157). Using quantitative Real-Time PCR, we evaluated expression of the selected Populus NACs in a developmental gradient and in response to bending stress. We prepared transgenic Arabidopsis and Populus plants with increased or decreased expression of select NAC genes. For dominant repression of target gene expression, we evaluated transgenic plants expressing translational fusions of NAC-EAR (ERF amphiphilic repressor) chimeras through chimeric repressor silencing-technology (CRES-T). XND1 overexpression in Populus and Arabidopsis resulted in severe stunting and suppression of xylem differentiation. Overexpression of PopNAC122, an XND1 ortholog, yielded an analogous phenotype in Arabidopsis. Populus XND1 overexpressors lacked phloem fibers and showed a reduction in cell size and number, vessel number and frequency of rays. Knowledge gained through characterization of these wood-associated regulatory genes can be used to optimize molecular breeding and genetic engineering strategies for improved wood quality and increased biomass.
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    Functional Diversification of Populus FLOWERING LOCUS D-LIKE3 Transcription Factor and Two Paralogs in Shoot Ontogeny, Flowering, and Vegetative Phenology
    Sheng, Xiaoyan; Hsu, Chuan-Yu; Ma, Cathleen; Brunner, Amy M. (Frontiers, 2022-02-03)
    Both the evolution of tree taxa and whole-genome duplication (WGD) have occurred many times during angiosperm evolution. Transcription factors are preferentially retained following WGD suggesting that functional divergence of duplicates could contribute to traits distinctive to the tree growth habit. We used gain- and loss-of-function transgenics, photoperiod treatments, and circannual expression studies in adult trees to study the diversification of three Populus FLOWERING LOCUS D-LIKE (FDL) genes encoding bZIP transcription factors. Expression patterns and transgenic studies indicate that FDL2.2 promotes flowering and that FDL1 and FDL3 function in different vegetative phenophases. Study of dominant repressor FDL versions indicates that the FDL proteins are partially equivalent in their ability to alter shoot growth. Like its paralogs, FDL3 overexpression delays short day-induced growth cessation, but also induces distinct heterochronic shifts in shoot development-more rapid phytomer initiation and coordinated delay in both leaf expansion and the transition to secondary growth in long days, but not in short days. Our results indicate that both regulatory and protein coding sequence variation contributed to diversification of FDL paralogs that has led to a degree of specialization in multiple developmental processes important for trees and their local adaptation.
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    Genome-wide analysis of Aux/IAA and ARF gene families in Populus trichocarpa
    Kalluri, Udaya C.; DiFazio, Stephen P.; Brunner, Amy M.; Tuskan, Gerald A. (2007-11-06)
    Background Auxin/Indole-3-Acetic Acid (Aux/IAA) and Auxin Response Factor (ARF) transcription factors are key regulators of auxin responses in plants. We identified the suites of genes in the two gene families in Populus and performed comparative genomic analysis with Arabidopsis and rice. Results A total of 35 Aux/IAA and 39 ARF genes were identified in the Populus genome. Comparative phylogenetic analysis revealed that several Aux/IAA and ARF subgroups have differentially expanded or contracted between the two dicotyledonous plants. Activator ARF genes were found to be two fold-overrepresented in the Populus genome. PoptrIAA and PoptrARF gene families appear to have expanded due to high segmental and low tandem duplication events. Furthermore, expression studies showed that genes in the expanded PoptrIAA3 subgroup display differential expression. Conclusion The present study examines the extent of conservation and divergence in the structure and evolution of Populus Aux/IAA and ARF gene families with respect to Arabidopsis and rice. The gene-family analysis reported here will be useful in conducting future functional genomics studies to understand how the molecular roles of these large gene families translate into a diversity of biologically meaningful auxin effects.
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    Growth and Physiological Responses to Fertilizer Application in Clonal Loblolly Pine
    Stovall, Jeremy Patrick (Virginia Tech, 2010-06-16)
    More than 20 million clonal loblolly pines have been planted throughout the southeastern United States. Fertilizer has been applied to more than 6.5 million hectares of plantations to alleviate deficiencies of nitrogen and phosphorus that limit growth. Because cloning loblolly pine in large numbers has only become possible in the last decade, it is unknown how clones may respond differently to fertilizer application. Growth, growth efficiency, and biomass partitioning responses to fertilizer application were investigated among 25 clones planted in the Virginia Piedmont. Closely related clones varied in their fertilizer stem volume responses, but not enough to be statistically significant (p = 0.11). Clones varied in growth efficiency and partitioning to individual tissues, but clone-by-fertilizer interactions were not observed. Clonal variability was observed in root morphology, and maximum rooting depth showed a significant clone-by-fertilizer interaction. Clones with rapid growth rates can be selected with a range of other desirable traits. Short-term (i.e. weeks) responses to fertilization are often inconsistent with long-term (i.e. years) responses, but are critical to understanding growth responses. We investigated carbon allocation in two full-sibling clones of loblolly pine under two levels of fertilizer application over four months in a greenhouse. Using monthly harvests of some trees and ecophysiological measurements throughout, we determined carbon allocation on a monthly scale. In response to fertilizer application, both clones reduced allocation belowground and increased allocation to foliage to some extent, increasing whole-canopy photosynthetic capacity. However, these changes in allocation were ephemeral. By the end of the experiment, root-shoot ratios were no longer significantly affected by fertilizer application. Clones had allocation patterns distinct from one another, with one allocating more belowground and the other allocating more to stem mass. While their overall growth responses to fertilizer application were similar, the physiological mechanisms that resulted in these responses were different between clones. Results of the two studies indicate that while fertilizer responses may not need to be included when testing clones for deployment, knowledge of the fertilizer responses of widely-deployed clones would offer forest managers opportunities to apply clone-specific precision-silvicultural systems to optimize growth rates and manage for a range of products.
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    Identification and characterization of a matrix metalloproteinase (Pta1-MMP) expressed during Loblolly pine (Pinus taeda) seed development and germination
    Ratnaparkhe, Supriya M. (Virginia Tech, 2009-03-19)
    Extracellular matrix (ECM) modifications occur during plant growth, development, and in response to environmental stimuli. Key modulators of ECM modification in vertebrates, the extracellular matrix metalloproteinases (MMPs), have also been described in a few plants. Here, we report the identification of Loblolly pine (Pinus taeda) Pta1-MMP and its characterization during seed development and germination. The Pta1-MMP protein has the structural characteristics of other plant MMPs, and a recombinant protein (rPta-MMP) generated by using EST sequences for a seed-expressed MMP exhibits Zn2+-dependent protease activity, and is inhibited by the active site-binding hydroxamate inhibitor GM6001 and EDTA. The Pta1-MMP gene is expressed during embryo development, with transcript levels increasing from proembryo to early cotyledonary stage, then declining during late cotyledonary expansion and maturation drying. Protein extracts exhibited similar developmental-stage MMP-like activity. Seed imbibition in water facilited germination, which was stimulated by GA3 and inhibited by ABA. The timing of germination was mirrored by the presence of MMP-like protease activity in both water- and GA3-imbibed embryos. Pta1-MMP transcript levels increased in association with germination for both GA3- and water-treated embryos, in agreement with MMP-like activity. In contrast, by 10 days after imbibition, Pta1-MMP transcripts in ABA-treated embryos were at levels similar to the other treatments, although MMP-like activity was not observed. The application of GM6001 during Loblolly pine seed imbibition inhibited germination in a dose-dependent manner. Our results suggest that Pta1-MMP is required for ECM modification, facilitating the cell division and expansion required for both embryo development and germination. To our knowledge, this is the first report of an MMP in any gymnosperm and also its involvement in embryo development and subsequent germination.
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    Metabolic engineering of the pterin branch of folate synthesis by over-expression of a GTP cyclohydrolase I in peanut
    Juba, Nicole Czarina (Virginia Tech, 2011-10-14)
    Folate, also known as vitamin B9, is an essential dietary vitamin that provides the donor group for one carbon transfer reactions. Deficiency in folate is associated with neural tube birth defects (NTDs), cancer, cardiovascular disease, and anemia. In the US enriched food products including bread, pasta, and cereal are fortified with folic acid, the synthetic analog of folate. While effective in reducing NTDs, this practice is costly and not economically practical in developing countries. Folate biofortification, increasing the natural folate level in foods by metabolic engineering, has been proposed as a sustainable alternative to food fortification with folic acid. To increase folate levels in peanut seed, GTP cyclohydrolase I from Arabidopsis thaliana (AtGCHI) was introduced into peanut by biolistic transformation. Plant transformation vectors were constructed using publicly available or licensable vector components to avoid intellectual property restrictions that hinder commercialization. Thirteen peanut cultivars were evaluated for transformation efficiencies and regeneration potential. Expression levels of the AtGCHI transgene were determined by quantitative real-time PCR. The endogenous peanut GCHI (AhGCHI) was isolated and sequenced. Studies were conducted to test whether heterologous over-expression of AtGCHI altered expression of the endogenous AhGCHI. Seed-specific expression of AtGCHI does not affect AhGCHI transcript accumulation. For validation of the proposed folate biofortification strategy, vitamin quantification will be required. A liquid chromatography tandem mass spectrometry (LC/MS/MS) method was developed to identify and quantify the different forms of folate. However, additional work will be needed to determine sensitivity of the instrument, to optimize vitamin extraction, and to increase sufficient seed for vitamin extraction and analysis. Peanut products derived from folate biofortified peanut kernels will have a niche market in the United States, but there is a larger global implication as a mechanism for sustainable delivery of essential vitamins to populations that can not adopt synthetic vitamin supplementation/fortification. Successful demonstration of increased folate in peanut will result in better vitamin availability for populationssonsuming peanut based foods as a dietary staple.
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    Overexpression of SHORT VEGETATIVE PHASE-LIKE (SVL) in Populus delays onset and reduces abundance of flowering in field-grown trees
    Goralogia, Greg S.; Howe, Glenn T.; Brunner, Amy M.; Helliwell, Emily; Nagle, Michael F.; Ma, Cathleen; Lu, Haiwei; Goddard, Amanda L.; Magnuson, Anna C.; Klocko, Amy L.; Strauss, Steven H. (2021-12)
    The spread of transgenes and exotic germplasm from planted crops into wild or feral species is a difficult problem for public and regulatory acceptance of genetically engineered plants, particularly for wind-pollinated trees such as poplar. We report that overexpression of a poplar homolog of the floral repressor SHORT VEGETATIVE PHASE-LIKE (SVL), a homolog of the Arabidopsis MADS-box repressor SHORT VEGETATIVE PHASE (SVP), delayed the onset of flowering several years in three genotypes of field-grown transgenic poplars. Higher expression of SVL correlated with a delay in flowering onset and lower floral abundance, and did not cause morphologically obvious or statistically significant effects on leaf characteristics, tree form, or stem volume. Overexpression effects on reproductive and vegetative phenology in spring was modest and genotype-specific. Our results suggest that use of SVL and related floral repressors can be useful tools to enable a high level of containment for vegetatively propagated short-rotation woody energy or pulp crops.
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    Partitioning of multivariate phenotypes using regression trees reveals complex patterns of adaptation to climate across the range of black cottonwood (Populus trichocarpa)
    Oubida, Regis Wendpouire (Virginia Tech, 2014-03-04)
    Local adaptation to climate in temperate forest trees involves the integration of multiple physiological, morphological, and phenological traits. Latitudinal clines for the relevant component traits are frequently observed for species that have a north-south distribution, but these relationships do not account for climatic variation within a given latitudinal band, which may be reflected in adaptive traits. We used black cottonwood (Populus trichocarpa) as a model to characterize the interplay between geography, climate, and adaptation to abiotic factors. Twelve traits (height, diameter, volume index, crown diameter, number of branches, number of sylleptic branches, relative number of branches, Relative canopy depth, Bud set, Bud flush, cold index of injury, carbon isotope ratio) were measured in a range-wide sample of 124 P. trichocarpa genotypes grown in a common garden. Heritability's were moderate to high (0.24 to 0.55) and significant population differentiation (QST > 0.3) suggested adaptive divergence. When climate variables were taken as predictors and the 12 traits as response variables in a multivariate regression tree analysis, aridity (Eref) explained the most variation, with subsequent splits grouping individuals according to mean temperature of the warmest month, frost-free period (FFP), and mean annual precipitation (MAP). This grouping matches relatively well the splits using geographic variables as predictors: the northernmost groups (short FFP and low Eref) had the lowest growth performance, and the highest cold hardiness. The groups spanning the south of British Columbia (low Eref and intermediate temperatures) displayed an average growth and cold hardiness. The group from the coast of California and Oregon (high Eref and FFP) had the best growth performance and the lowest cold hardiness. The southernmost and high-elevated group (with High Eref and low FFP) performed poorly, had a low cold hardiness and a significantly lower WUE.
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    Phenotypic Expression and Stability in a Large-Scale Field Study of Genetically Engineered Poplars Containing Sexual Containment Transgenes
    Klocko, Amy L.; Lu, Haiwei; Magnuson, Anna C.; Brunner, Amy M.; Ma, Cathleen; Strauss, Steven H. (2018-08-03)
    Genetic engineering (GE) has the potential to help meet demand for forest products and ecological services However, high research and development costs, market restrictions, and regulatory obstacles to performing field tests have severely limited the extent and duration of field research. There is a notable paucity of field studies of flowering GE trees due to the time frame required and regulatory constraints. Here we summarize our findings from field testing over 3,300 GE poplar trees and 948 transformation events in a single, 3.6 hectare field trial for seven growing seasons; this trial appears to be the largest field-based scientific study of GE forest trees in the world. The goal was to assess a diversity of approaches for obtaining bisexual sterility by modifying RNA expression or protein function of floral regulatory genes, including LEAFY, AGAMOUS, APETALA1, SHORT VEGETATIVE PHASE, and FLOWERING LOCUS T. Two female and one male clone were transformed with up to 23 different genetic constructs designed to obtain sterile flowers or delay onset of flowering. To prevent gene flow by pollen and facilitate regulatory approval, the test genotypes chosen were incompatible with native poplars in the area. We monitored tree survival, growth, floral onset, floral abundance, pollen production, seed formation and seed viability. Tree survival was above 95%, and variation in site conditions generally had a larger impact on vegetative performance and onset of flowering than did genetic constructs Floral traits, when modified, were stable over three to five flowering seasons, and we successfully identified RNAi or overexpression constructs that either postponed floral onset or led to sterile flowers. There was an absence of detectable somaclonal variation; no trees were identified that showed vegetative or floral modifications that did not appear to be related to the transgene added. Surveys for seedling and sucker establishment both within and around the plantation identified small numbers of vegetative shoots (root sprouts) but no seedlings, indicative of a lack of establishment of trees via seeds in the area. Overall, this long term study showed that GE containment traits can be obtained which are effective, stable, and not associated with vegetative abnormalities or somaclonal variation
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    Populus biomass protein-protein interactions and their functions
    Jia, Xiaoyan; Zhao, Mingzhe; Zhao, Chengsong; Sheng, Xiaoyan; Dickerman, Allan; Beers, Eric P.; Brunner, Amy M. (2011-09-13)
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    Regulation of shoot-system development in Populus
    Brunner, Amy M.; Sheng, Xiaoyan; Edwards, Joesph; Fujino, Takeshi; Wang, Chieh-Ting; DiFazio, Stephen P. (2011-09-13)
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    RNA interference suppression of AGAMOUS and SEEDSTICK alters floral organ identity and impairs floral organ determinacy, ovule differentiation, and seed-hair development in Populus
    Lu, Haiwei; Klocko, Amy L.; Brunner, Amy M.; Ma, Cathleen; Magnuson, Anna C.; Howe, Glenn T.; An, Xinmin; Strauss, Steven H. (2019-04)
    The role of the floral homeotic gene AGAMOUS (AG) and its close homologues in development of anemophilous, unisexual catkins has not previously been studied. We transformed two RNA interference (RNAi) constructs, PTG and its matrix-attachment-region flanked version MPG, into the early-flowering female poplar clone 6K10 (Populus alba) to suppress the expression of its two duplicate AG orthologues. By early 2018, six out of 22 flowering PTG events and 11 out of 12 flowering MPG events showed modified floral phenotypes in a field trial in Oregon, USA. Flowers in catkins from modified events had carpel-inside-carpel' phenotypes. Complete disruption of seed production was observed in seven events, and sterile anther-like organs in 10 events. Events with strong co-suppression of both the two AG and two SEEDSTICK (STK) paralogues lacked both seeds and associated seed hairs. Alterations in all of the modified floral phenotypes were stable over 4yr of study. Trees from floral-modified events did not differ significantly (P<0.05) from nonmodified transgenic or nontransgenic controls in biomass growth or leaf morphology. AG and STK genes show strong conservation of gene function during poplar catkin development and are promising targets for genetic containment of exotic or genetically engineered trees.
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    RNAi of AGAMOUS genes in sweetgum alters reproductive organ identity and decreases fruit persistence
    Klocko, Amy L.; Brunner, Amy M.; Ma, Cathleen; Etherington, Elizabeth; Rosenstiel, Kori; Magnuson, Anna C.; Taylor, Barbara J.; Cappellazzi, Jed; Lockwood, Thomas; Covarrubias, Nichole; Bao, Manzhu; Morrell, Jeffrey J.; Strauss, Steven H. (2020-05)
    Sweetgums (Liquidambar), members of the family Altingiaceae (Altingiales), have inflorescences and floral organs that are distinctive in structure compared with other angiosperms in which the roles of floral homeotic genes have been studied. To begin to understand the role of AGAMOUS (AG)-a floral homeotic gene that has a major role in stamen and carpel development-in development of the monosexual flowers of sweetgum, we used RNAi to reduce the expression of two members of the AG subfamily. Because AG suppression should induce floral sterility, RNAi might also provide a tool to mitigate the risks of invasiveness-and to reduce the production of its nuisance fruits or allergenic pollen-when sweetgum is used as an exotic shade or forest tree. We tested 33 independent transgenic events and non-transgenic controls during 10 years in the field. The RNAi-AG sweetgum trees maintained normal growth, phenology, and vivid fall coloration during the 10 years of study, but 8 insertion events had highly modified inflorescence and floral morphology. The modified flowers had anthers and carpels that were converted to flat leaf-like structures lacking pollen grains and ovules, respectively. The female inflorescences developed into dry papery structures that failed to produce seeds. These infructescences were smaller than control infructescences, and lost a greater percentage of biomass in a controlled decay assay. RNAi against AG genes was highly effective at impairing fertility and modifying reproductive development without significant vegetative effects in sweetgum and gave phenotypes distinct from, but similar to, that of AG loss of function in other angiosperms.
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    Strategies for Engineering Reproductive Sterility in Plantation Forests
    Fritsche, Steffi; Klocko, Amy L.; Boron, Agnieszka; Brunner, Amy M.; Thorlby, Glenn (2018-11-15)
    A considerable body of research exists concerning the development of technologies to engineer sterility in forest trees. The primary driver for this work has been to mitigate concerns arising from gene flow from commercial plantings of genetically engineered (GE) trees to non-GE plantations, or to wild or feral relatives. More recently, there has been interest in the use of sterility technologies as a means to mitigate the global environmental and socio-economic damage caused by the escape of non-native invasive tree species from planted forests. The current sophisticated understanding of the molecular processes underpinning sexual reproduction in angiosperms has facilitated the successful demonstration of a number of control strategies in hardwood tree species, particularly in the model hardwood tree Poplar. Despite gymnosperm softwood trees, such as pines, making up the majority of the global planted forest estate, only pollen sterility, via cell ablation, has been demonstrated in softwoods. Progress has been limited by the lack of an endogenous model system, long timescales required for testing, and key differences between softwood reproductive pathways and those of well characterized angiosperm model systems. The availability of comprehensive genome and transcriptome resources has allowed unprecedented insights into the reproductive processes of both hardwood and softwood tree species. This increased fundamental knowledge together with the implementation of new breeding technologies, such as gene editing, which potentially face a less oppressive regulatory regime, is making the implementation of engineered sterility into commercial forestry a realistic possibility.
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    Synergies and Entanglement in Secondary Cell Wall Development and Abiotic Stress Response in Trees
    Coleman, Heather D.; Brunner, Amy M.; Tsai, Chung-Jui (2021-03-19)
    A major challenge for sustainable food, fuel, and fiber production is simultaneous genetic improvement of yield, biomass quality, and resilience to episodic environmental stress and climate change. For Populus and other forest trees, quality traits involve alterations in the secondary cell wall (SCW) of wood for traditional uses, as well as for a growing diversity of biofuels and bioproducts. Alterations in wood properties that are desirable for specific end uses can have negative effects on growth and stress tolerance. Understanding of the diverse roles of SCW genes is necessary for the genetic improvement of fast-growing, short-rotation trees that face perennial challenges in their growth and development. Here, we review recent progress into the synergies and antagonisms of SCW development and abiotic stress responses, particularly, the roles of transcription factors, SCW biogenesis genes, and paralog evolution.
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    Transgenic Suppression of AGAMOUS Genes in Apple Reduces Fertility and Increases Floral Attractiveness
    Klocko, Amy L.; Borejsza-Wysocka, Ewa; Brunner, Amy M.; Shevchenko, Olga; Aldwinckle, Herb; Strauss, Steven H. (PLOS, 2016-08-08)
    We investigated the ability of RNA interference (RNAi) directed against two co-orthologs of AGAMOUS (AG) from Malus domestica (domestic apple, MdAG) to reduce the risks of invasiveness and provide genetic containment of transgenes, while also promoting the attractiveness of flowers for ornamental usage. Suppression of two MdAG-like genes, MdMADS15 and MdMADS22, led to the production of trees with highly showy, polypetalous flowers. These “double-flowers” had strongly reduced expression of both MdAG-like genes. Members of the two other clades within in the MdAG subfamily showed mild to moderate differences in gene expression, or were unchanged, with the level of suppression approximately proportional to the level of sequence identity between the gene analyzed and the RNAi fragment. The double-flowers also exhibited reduced male and female fertility, had few viable pollen grains, a decreased number of stigmas, and produced few viable seeds after cross-pollination. Despite these floral alterations, RNAi-AG trees with double-flowers set full-sized fruit. Suppression or mutation of apple AG-like genes appears to be a promising method for combining genetic containment with improved floral attractiveness.
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    Vernalization and the chilling requirement to exit bud dormancy: shared or separate regulation?
    Brunner, Amy M.; Evans, Luke M.; Hsu, Chuan-Yu; Sheng, Xiaoyan (Frontiers, 2014-12-17)
    Similarities have long been recognized between vernalization, the prolonged exposure to cold temperatures that promotes the floral transition in many plants, and the chilling requirement to release bud dormancy in woody plants of temperate climates. In both cases the extended chilling period occurring during winter is used to coordinate developmental events to the appropriate seasonal time. However, whether or not these processes share common regulatory components and molecular mechanisms remain largely unknown. Both gene function and association genetics studies in Populus are beginning to answer this question. In Populus, studies have revealed that orthologs of the antagonistic flowering time genes FT and CEN/TFL1 might have central roles in both processes. We review Populus seasonal shoot development related to dormancy release and the floral transition and evidence for FT/TFL1-mediated regulation of these processes to consider the question of regulatory overlap. In addition, we discuss the potential for and challenges to integrating functional and population genomics studies to uncover the regulatory mechanisms underpinning these processes in woody plant systems.