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At least 55 records · Page 3

Discovering Innovations in Stress Tolerance through Comparative Gene Regulatory Network Analysis and Cell-Type Specific Expression Maps (Final Technical Report with Cover Page)

Through this grant, we developed a comparative framework to elucidate the mechanisms behind variations in environmental stress responses among a diverse group of species within the Brassicaceae family. Our focus was on the differences in physiological and transcriptomic responses to abscisic acid (ABA), a hormone associated with water stress. We examined the differential growth responses of four Brassicaceae species, finding that most exhibited reduced root growth correlated with smaller meristem size. In contrast, Schrenkiella parvula showed accelerated growth due to increased root cell elongation. We employed RNA sequencing to analyze the transcriptional responses to ABA across these species, and innovative bioinformatics techniques were used to pinpoint biological pathways with significant divergence. Additionally, we utilized DAP-seq to map the gene regulatory networks associated with ABAresponsive transcription factors, revealing that variations in the regulation of growth hormone biosynthesis play a critical role in the distinct ABA effects on root growth among the species. This research sets a new standard for comparative physiology by integrating comparative genomics and transcriptomics to uncover pathway divergences.

59 BASIC BIOLOGICAL SCIENCES↗

Enhanced Resistance Pines for Improved Renewable Biofuel and Chemical Production (Technical Report)

We completed phenotyping constitutive and inducible oleoresin flow across two seasons, constitutive resin canal number and density and wood terpene content in our ADEPT2 and CCLONES populations. We completed genetic association between 19 oleoresin phenotypes and a total of 523,192 SNP markers from ADEPT2 and 13,883 SNP markers in CCLONES using four mixed linear models. A total of 293 significant SNPs (FDR = 0.20) were identified. We used the MENTOR tool to mine mechanistic connections from a multiplex network constructed from poplar multi-omic data to construct a conceptual model for a subset of these significant SNPs. Our model contains 6 transcriptional regulators in addition to 3 monoterpene synthases. To generate more lines of evidence for these significant SNPs, we completed a time course RNAseq experiment after inducing vascular zone cells to differentiate into new resin canals with a methyl jasmonate treatment, a single nuclei RNAseq that identified differentiating resin canal epithelial cells and are completing analysis for a QTL study in a hybrid pine population. The time course identified 4634 significantly down and 1890 significantly up regulated transcripts after treatment with methyl jasmonate, an inducer of new resin canal formation in the vascular cambial meristem. To analyze this large set of differentially regulated genes, we created a predictive expression network and analyzed it with random walk restart using 6 seed genes coding for transcription factors regulating xylem differentiation in poplar. Of the top ranked 200 transcripts, 119 transcripts were significant differentially expressed supporting these transcripts as potential candidates regulating resin canal formation. Analysis of single nuclei sequencing of shoot tips that contain differentiating resin canals, identified 10 clusters. One cluster was highly enriched in transcripts coding for 9 of the enzymes in the MEP pathway 3 prenyl synthetases, and 3 monoterpene synthases strongly suggesting that this cluster represents resin canal epithelial cells. We are mining the additional transcripts to create a trajectory analysis. In summary, we have identified > 10 novel genes that are strongly supported candidates for further analysis in breeding lines and for genetic engineering over- and under- expressing lines to increase wood terpene content to improve resistance to insect and fungal pathogens while simultaneously increasing terpene supplies for renewable chemicals and biofuels.

59 BASIC BIOLOGICAL SCIENCES↗

Annual Herbaceous Plants Exhibit Altered Morphological Traits in Response to Altered Precipitation and Drought Patterns in Semiarid Sandy Grassland, Northern China

The frequency and intensity of extreme precipitation events and severe drought are predicted to increase in semiarid areas due to global climate change. Plant morphological traits can reflect plant responses to a changing environment, such as altered precipitation or drought patterns. In this study, we examined the response of morphological traits of root, stem, leaf and reproduction meristems of annual herbaceous species to altered precipitation and drought patterns in a semiarid sandy grassland. The study involved a control treatment (100% of background precipitation) and the following six altered precipitation treatments: (1) P(+): precipitation increased by 30%, (2) P(++): precipitation increased by 60%, (3) P(-): precipitation decreased by 30%, (4) P(--): precipitation decreased by 60%, (5) drought 1 (D1): 46-day drought from May 1st to June 15th, and (6) drought 2 (D2): 46-day drought from July 1st to August 15th. P(++) significantly increased root length, flower length-to-width ratio, both P(+) and P(++) significantly increased stem length and flower number in the plant growing seasons, while all of them decreased under P(-) and P(--). The annual herbaceous plants marginally increased the number of second-level stem branches and stem diameter in order to better resist the severe drought stress under P(--). P(+) and P(++) increased the root, stem, leaf, and flower dry weight, with the flower dry weight accounting for a larger proportion than the other aboveground parts. Under D2, the plants used the limited water resources more efficiently by increasing the root-to-shoot ratio compared with P(-), P(--) and D1, which reflects biomass allocation to belowground increased. The linear mixed-effects models and redundancy analysis showed that the root-to-shoot ratio and the dry weight of various plant components were significantly affected by morphological traits and altered precipitation magnitude. Our results showed that the herbaceous species have evolved morphological trait responses that allow them to adapt to climate change. Such differences in morphological traits may ultimately affect the growing patterns of annual herbaceous species, enhancing their drought-tolerant capacity in semiarid sandy grassland during the ongoing climate change.

Sun, Shan-Shan↗

Non-TZF Protein AtC3H59/ZFWD3 Is Involved in Seed Germination, Seedling Development, and Seed Development, Interacting with PPPDE Family Protein Desi1 in Arabidopsis

Despite increasing reports on the function of CCCH zinc finger proteins in plant development and stress response, the functions and molecular aspects of many non-tandem CCCH zinc finger (non-TZF) proteins remain uncharacterized. AtC3H59/ZFWD3 is an Arabidopsis non-TZF protein and belongs to the ZFWD subfamily harboring a CCCH zinc finger motif and a WD40 domain. In this study, we characterized the biological and molecular functions of AtC3H59, which is subcellularly localized in the nucleus. The seeds of AtC3H59-overexpressing transgenic plants (OXs) germinated faster than those of wild type (WT), whereas atc3h59 mutant seeds germinated slower than WT seeds. AtC3H59 OX seedlings were larger and heavier than WT seedlings, whereas atc3h59 mutant seedlings were smaller and lighter than WT seedlings. Moreover, AtC3H59 OX seedlings had longer primary root length than WT seedlings, whereas atc3h59 mutant seedlings had shorter primary root length than WT seedlings, owing to altered cell division activity in the root meristem. During seed development, AtC3H59 OXs formed larger and heavier seeds than WT. Using yeast two-hybrid screening, we isolated Desi1, a PPPDE family protein, as an interacting partner of AtC3H59. AtC3H59 and Desi1 interacted via their WD40 domain and C-terminal region, respectively, in the nucleus. Taken together, our results indicate that AtC3H59 has pleiotropic effects on seed germination, seedling development, and seed development, and interacts with Desi1 in the nucleus via its entire WD40 domain. To our knowledge, this is the first report to describe the biological functions of the ZFWD protein and Desi1 in Arabidopsis.

59 BASIC BIOLOGICAL SCIENCES↗

Role of SaPCR2 in Zn Uptake in the Root Elongation Zone of the Zn/Cd Hyperaccumulator Sedum alfredii

Zn pollution is a potential toxicant for agriculture and the environment. Sedum alfredii is a Zn/Cd hyperaccumulator found in China and has been proven as a useful resource for the phytoremediation of Zn-contaminated sites. However, the molecular mechanism of Zn uptake in S. alfredii is limited. In this study, the function of SaPCR2 on Zn uptake in S. alfredii was identified by gene expression analysis, yeast function assays, Zn accumulation and root morphology analysis in transgenic lines to further elucidate the mechanisms of uptake and translocation of Zn in S. alfredii. The results showed that SaPCR2 was highly expressed in the root elongation zone of the hyperaccumulating ecotype (HE) S. alfredii, and high Zn exposure downregulated the expression of SaPCR2 in the HE S. alfredii root. The heterologous expression of SaPCR2 in yeast suggested that SaPCR2 was responsible for Zn influx. The overexpression of SaPCR2 in the non-hyperaccumulating ecotype (NHE) S. alfredii significantly increased the root uptake of Zn, but did not influence Mn, Cu or Fe. SR-μ-XRF technology showed that more Zn was distributed in the vascular buddle tissues, as well as in the cortex and epidermis in the transgenic lines. Root morphology was also altered after SaPCR2 overexpression, and a severe inhibition was observed. In the transgenic lines, the meristematic and elongation zones of the root were lower compared to the WT, and Zn accumulation in meristem cells was also reduced. These results indicate that SaPCR2 is responsible for Zn uptake, and mainly functions in the root elongation zone. This research on SaPCR2 could provide a theoretical basis for the use of genetic engineering technology in the modification of crops for their safe production and biological enhancement.

59 BASIC BIOLOGICAL SCIENCES↗

Plant Adaptation to Cold. 1: Chlorophyll. 2: Minerals

A number of montane herbs in northern Utah typically form flower buds beneath the snow cover and flower either through it or immediately after its recession. Two of these species, one naturally occurring, Claytonia lanceolata, and one cultivated bulb, Galanthus nivalis, were investigated for their response to this stress environment. Snow depth patterns, chlorophyll content of tissues, and plants grown in light-tight boxes, suggest that light passing through the snow to reach plants growing underneath is not critically involved in the timing of their developmental cycles or in their ability to endure this low temperature environment. Ability to endure stress seems to be closely related in a number of ways to activity at the plant membranes. Plants were protected from low temperature damage by application of cytokinin or calcium, both of which probably acted at the membrane. Potassium calcium antagonisms were reflected in the internal distribution of the ions under natural stress conditions; and plants that differentiated at the meristem while growing through the snow accumulated calcium at the tip during this growth.

Rosen, P.↗

Genetic changes induced by space flight factors in barley seeds on Soyuz-5 and Soyuz-9 craft

Air-dry seeds of the barley Zimujuschij moscowskyi of the 1969 harvest were taken into space onboard the spaceships Soyuz-5 and Soyuz-9. A cytological study of the mitoses in meristemic cells in rootlet terminals revealed that space flight factors (SFF) in nonirradiated seeds induced about 3% of aberrant cells. After irradiation the effect of SFF increased over two-fold. Although the radio protectors ensured the seeds against from the SFF-induced damage either in irradiated or nonirradiated seed cells which is inconsistent with the previously obtained data.

Nuzhdin, N. I.↗

Calcium elicited asymmetric auxin transport in gravity influenced root segments

Auxin is a prime candidate for regulating and modulating the differential growth response of primary corn roots to gravity. Auxin, indole-3-acetic acid (IAA), both promotes and inhibits root elongation rapidly within a narrow concentration range. Thus growth regulation would require only small changes in the short lag period for initiation of gravitropism. Since auxin is transported to/through the zone of elongation toward the meristem, it may serve as a direct communication link between the zone of elongation, site of gravitropic response, and the root cap (RC), site of gravity perception. When auxin transport is inhibited, gravitropism is also inhibited. Napthylpthalamic acid (NPA) is one such inhibitor. It inhibits gravitropism only when applied to the apical growing and dividing region of the root. Application at the basal end of the root does not influence gravitropic NPA causes upward curvature when applied to the upper surface of horizontal, two day-old, intact corn roots. This effect is countered by application of IAA to the opposite side.

Edwards, K. L.↗

Transduction of the Root Gravitropic Stimulus: Can Apical Calcium Regulate Auxin Distribution?

The hypothesis was tested that calcium, asymmetrically distributes in the root cap upon reorientation to gravity, affects auxin transport and thereby auxin distribution at the elongation zone. It is assumed that calcium exists in the root cap and is asymmetrically transported in root caps altered from a vertical to a horizontal position and that the meristem, the tissue immediately adjacent to the root cap and lying between the site of gravity perception and the site of gravity response, is essential for mediation of gravitropism. Tip calcium in root gravicurvature was implicated. The capstone evidence is that the root cap has the capacity to polarly translocate exogenous calcium downward when tissue is oriented horizontally, and that exogenous calcium, when supplied asymmetrically at the root tip, induces curvature and dictates the direction of curvature in both vertical and horizontal corn roots.

Edwards, K. L.↗

Modification of cytogenetic and physiological effects of space flight factors by biologically active compounds

Physiological and cytogenetic changes in the Welsh onion plants induced by a short (82 days) and long term (522 days) space flight are expressed in decrease of seed germination, inhibition of stem growth, depression of cell division in root meristem, and increase in the number of structural chromosome rearrangements. The treatment of such plants with solutions of a-tocopherol, auxin, and kinetin decreased the level of chromosome aberrations to the control one and normalized cell divisions and growth partly or completely.

Aliyev, A. A.↗

Root gravitropism in maize and Arabidopsis

Research during the period 1 March 1992 to 30 November 1993 focused on improvements in a video digitizer system designed to automate the recording of surface extension in plants responding to gravistimulation. The improvements included modification of software to allow detailed analysis of localized extension patterns in roots of Arabidopsis. We used the system to analyze the role of the postmitotic isodiametric growth zone (a region between the meristem and the elongation zone) in the response of maize roots to auxin, calcium, touch and gravity. We also used the system to analyze short-term auxin and gravitropic responses in mutants of Arabidopsis with reduced auxin sensitivity. In a related project, we studied the relationship between growth rate and surface electrical currents in roots by examining the effects of gravity and thigmostimulation on surface potentials in maize roots.

Evans, Michael L.↗

Cosmic heavy ion tracks in mesoscopic biological test objects

Since more than 20 years ago, when the National Academy of Sciences and the National Research Council of the U.S.A. released their report on 'HZE particle effects in manned spaced flight', it has been emphasized how difficult - if not even impossible - it is to assess their radiobiological impact on man from conventional studies where biological test organisms are stochastically exposed to 'large' fluences of heavy ions. An alternative, competing approach had been realized in the BIOSTACK experiments, where the effects of single cosmic as well as accelerator - heavy ions on individual biological test organisms could be investigated. Although presented from the beginning as the preferable approach for terrestrial investigations with accelerator heavy ions too ('The BIOSTACK as an approach to high LET radiation research'), only recently this insight is gaining more widespread recognition. In space flight experiments, additional constraints imposed by the infrastructure of the vehicle or satellite further impede such investigations. Restrictions concern the physical detector systems needed for the registration of the cosmic heavy ions' trajectories as well as the biological systems eligible as test organisms. Such optimized procedures and techniques were developed for the investigations on chromosome aberrations induced by cosmic heavy ions in cells of the stem meristem of lettuce seeds (Lactuca sativa) and for the investigation of the radiobiological response of Wolffia arriza, which is the smallest flowering (water) plant. The biological effects were studied by the coworkers of the Russian Institute of Biomedical Problems (IBMP) which in cooperation with the European Space Agency ESA organized the exposure in the Biosatellites of the Cosmos series. Since biological investigations and physical measurements of particle tracks had to be performed in laboratories widely separated, the preferred fixed contact between biological test objects and the particle detectors until the geometrical correlation between tracks and organisms has been established could not be maintained. This gave rise to half a dozen of coordinate systems for different measurements which finally had to be related to a single stack reference system.

Facius, R.↗

Induced Abnormality In Mir- and Earth-Grown Super Dwarf Wheat

Super-dwarf wheat grown on the Mir space station using the Svet "Greenhouse" exhibited morphological, metabolic and reproductive abnormalities compared with normal wheat. Of prominent importance were the abnormalities associated with reproductive ontogeny and the total absence of seed formation on Mir. Changes in the apical meristem associated with transition from the vegetative phase to floral initiation and development of the reproductive spike were all typical of 'Super Dwarf' wheat up to the point of anthesis. Observation of ruptured anthers from the Mir-grown plants revealed what appeared to be normally developed pollen. These pollen grains however, contain only one nucleus, while normal viable pollen is trinucleate. A potentially important difference in the flight experiment, compared with ground reference studies, was identified - a high level of atmospheric ethylene (1200 ppb). Ground studies conducted exposing "Super-dwarf" wheat to ethylene at just prior to anthesis resulted in manifestation of the same abnormalities observed in the space flight samples.

Bubenheim, David L.↗

Transport of the two natural auxins, indole-3-butyric acid and indole-3-acetic acid, in Arabidopsis

Polar transport of the natural auxin indole-3-acetic acid (IAA) is important in a number of plant developmental processes. However, few studies have investigated the polar transport of other endogenous auxins, such as indole-3-butyric acid (IBA), in Arabidopsis. This study details the similarities and differences between IBA and IAA transport in several tissues of Arabidopsis. In the inflorescence axis, no significant IBA movement was detected, whereas IAA is transported in a basipetal direction from the meristem tip. In young seedlings, both IBA and IAA were transported only in a basipetal direction in the hypocotyl. In roots, both auxins moved in two distinct polarities and in specific tissues. The kinetics of IBA and IAA transport appear similar, with transport rates of 8 to 10 mm per hour. In addition, IBA transport, like IAA transport, is saturable at high concentrations of auxin, suggesting that IBA transport is protein mediated. Interestingly, IAA efflux inhibitors and mutations in genes encoding putative IAA transport proteins reduce IAA transport but do not alter IBA movement, suggesting that different auxin transport protein complexes are likely to mediate IBA and IAA transport. Finally, the physiological effects of IBA and IAA on hypocotyl elongation under several light conditions were examined and analyzed in the context of the differences in IBA and IAA transport. Together, these results present a detailed picture of IBA transport and provide the basis for a better understanding of the transport of these two endogenous auxins.

Non-NASA Center↗

Induced abnormality in Mir- and Earth grown Super Dwarf wheat

Super-dwarf wheat grown on the Mir space station using the Svet "Greenhouse" exhibited morphological, metabolic and reproductive abnormalities compared with Earth-grown wheat. Of prominent importance were the abnormalities associated with reproductive ontogeny and the total absence of seed formation on Mir. Changes in the apical meristem associated with transition from the vegetative phase to floral initiation and development of the reproductive spike were all typical of 'Super-Dwarf' wheat up to the point of anthesis. Observation of ruptured anthers from the Mir-grown plants revealed what appeared to be normally developed pollen. These pollen gains, however, contained only one nuclei, while normal viable pollen is tri-nucleate. A potentially important difference in the flight experiment, compared with ground reference studies, was the presence of a high level of atmospheric ethylene (1,200 ppb). Ground studies conducted by exposing 'Super-Dwarf' wheat to ethylene just prior to anthesis resulted in manifestation of the same abnormalities observed in the space flight samples. c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

long duration↗

Genes encoding calmodulin-binding proteins in the Arabidopsis genome

Analysis of the recently completed Arabidopsis genome sequence indicates that approximately 31% of the predicted genes could not be assigned to functional categories, as they do not show any sequence similarity with proteins of known function from other organisms. Calmodulin (CaM), a ubiquitous and multifunctional Ca(2+) sensor, interacts with a wide variety of cellular proteins and modulates their activity/function in regulating diverse cellular processes. However, the primary amino acid sequence of the CaM-binding domain in different CaM-binding proteins (CBPs) is not conserved. One way to identify most of the CBPs in the Arabidopsis genome is by protein-protein interaction-based screening of expression libraries with CaM. Here, using a mixture of radiolabeled CaM isoforms from Arabidopsis, we screened several expression libraries prepared from flower meristem, seedlings, or tissues treated with hormones, an elicitor, or a pathogen. Sequence analysis of 77 positive clones that interact with CaM in a Ca(2+)-dependent manner revealed 20 CBPs, including 14 previously unknown CBPs. In addition, by searching the Arabidopsis genome sequence with the newly identified and known plant or animal CBPs, we identified a total of 27 CBPs. Among these, 16 CBPs are represented by families with 2-20 members in each family. Gene expression analysis revealed that CBPs and CBP paralogs are expressed differentially. Our data suggest that Arabidopsis has a large number of CBPs including several plant-specific ones. Although CaM is highly conserved between plants and animals, only a few CBPs are common to both plants and animals. Analysis of Arabidopsis CBPs revealed the presence of a variety of interesting domains. Our analyses identified several hypothetical proteins in the Arabidopsis genome as CaM targets, suggesting their involvement in Ca(2+)-mediated signaling networks.

NASA Discipline Plant Biology↗

Cellular specificity of the gravitropic motor response in roots

A number of features of the gravitropic response of roots are not readily accounted for by the classical Cholodny-Went theory. These include the observations that (i) in the later stages of the response the growth gradient is reversed with no evident reversal of the auxin gradient; (ii) a major component of the acceleration of growth along the upper side occurs in the distal elongation zone (DEZ), a group of cells located between the meristem and the main elongation, not within the central elongation zone; and (iii) the initiation of differential growth in the DEZ appears to be independent of the establishment of auxin asymmetry. Alternative candidates for mediation of differential growth in the DEZ include calcium ions and protons. Gravi-induced curvature is accompanied by polar movement of calcium toward the lower side of the maize root tip and the DEZ is shown to be particularly sensitive to growth inhibition by calcium. Also, gravistimulation of maize roots causes enhanced acid efflux from the upper side of the DEZ. Evidence for gravi-induced modification of ion movements in the root tip includes changes in intracellular potentials and current flow. It is clear that there is more than one motor region in the root with regard to gravitropic responses and there is evidence that the DEZ itself consists of more than one class of responding cells. In order to gain a more complete understanding of the mechanism of gravitropic curvature, the physiological properties of the sub-zones of the root apex need to be thoroughly characterized with regard to their sensitivity to hormones, calcium, acid pH and electrical perturbations.

NASA Discipline Plant Biology↗

Specialized zones of development in roots

The authors propose using the term "distal elongation zone" (DEZ) rather than "postmitotic isodiametric growth zone" to refer to the group of cells between the apical meristem and the elongation zone in plant roots. Reasons presented for the change are that the proposed DEZ includes many cells that are still dividing, most cells in the region are not isodiametric, and the pattern of cell expansion in this region varies with position in the region. Cells in the DEZ respond to gravistimulation, mechanical impedance, electrotropic stimulation, water stress, and auxin. Differences in gene expression patterns between DEZ cells and cells in the main elongation zone are noted.

NASA Discipline Plant Biology↗