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At least 19 records

Data from: "Lab and Field Warming Similarly Advance Germination Date and Limit Germination Rate for High and Low Elevation Provenances of Two Widespread Subalpine Conifers"

This archive contains data used to draw conclusions in “Lab and Field Warming Similarly Advance Germination Date and Limit Germination Rate for High and Low Elevation Provenances of Two Widespread Subalpine Conifers”, by Kueppers et al. 2017. Cone collection and field experiments were conducted on Niwot Ridge, Colorado, with the latter completed within the Alpine Treeline Warming Experiment (ATWE). Laboratory germination experiments were conducted at the University of California Merced. Geospatial files are also provided for additional geographical context.There are four data file formats in this package: comma-separated values (.csv), Microsoft Excel (.xlsx), keyhole markup language (.kml), and ESRI shapefile (.shp). The latter two are geospatial file formats. .csv files can be opened using any simple text editor program such as TextEdit (Mac) and Notepad (Windows), and .xlsx files can be opened using Microsoft Excel or Google Sheets. The .kml file contains coordinate data, and is compatible with Google Earth and Google Maps. ESRI shapefiles (.shp) are compatible with any geospatial software able to read the file type, such as QGIS and ESRI’s ArcGIS Suite. This data user’s guide is available in as a .pdf, and can be opened using software such as Adobe Acrobat Reader.-----------------------------------------------------------------------------------------------------------------------------------Accurately predicting upslope shifts in subalpine tree ranges with warming requires understanding how future forest populations will be affected by climate change, as these are the seed sources for new tree line and alpine populations. Early life history stages are particularly sensitive to climate and are also influenced by genetic variation among populations. We tested the climate sensitivity of germination and initial development for two widely distributed subalpine conifers, using controlled-environment growth chambers with one temperature regime from subalpine forest in the Colorado Rocky Mountains and one 5 °C warmer, and two soil moisture levels. We tracked germination rate and timing, rate of seedling development, and seedling morphology for two seed provenances separated by ~300 m elevation. Warming advanced germination timing and initial seedling development by a total of ~2 weeks, advances comparable to mean differences between provenances. Advances were similar for both provenances and species; however, warming reduced the overall germination rate, as did low soil moisture, only for Picea engelmannii. A three-year field warming and watering experiment planted with the same species and provenances yielded responses qualitatively consistent with the lab trials. Together these experiments indicate that in a warmer, drier climate, P. engelmannii germination, and thus regeneration, could decline, which could lead to declining subalpine forest populations, while Pinus flexilis forest populations could remain robust as a seed source for upslope range shifts.

54 ENVIRONMENTAL SCIENCES↗

Gene activity during germination of spores of the fern, Onoclea sensibilis. Cell-free translation analysis of mRNA of spores and the effect of alpha-amanitin on spore germination

Poly(A)-RNA fractions of dormant, dark-imbibed (non-germinating) and photoinduced (germinating) spores of Onoclea sensibilis were poor templates in the rabbit reticulocyte lysate protein synthesizing system, but the translational efficiency of poly(A)+RNA was considerably higher than that of unfractionated RNA. Poly(A)+RNA isolated from photoinduced spores had a consistently higher translational efficiency than poly(A)+RNA from dark-imbibed spores. Analysis of the translation products by one-dimensional polyacrylamide gel electrophoresis showed no qualitative differences in the mRNA populations of dormant, dark-imbibed, and photoinduced spores. However, poly(A)+RNA from dark-imbibed spores appeared to encode in vitro fewer detectable polypeptides at a reduced intensity than photoinduced spores. A DNA clone encoding the large subunit of maize ribulose bisphosphate carboxylase hybridized at strong to moderate intensity to RNA isolated from dark-imbibed spores, indicating the absence of mRNA degradation. Although alpha-amanitin did not inhibit the germination of spores, the drug prevented the elongation of the rhizoid and protonemal initial with a concomitant effect on the synthesis of poly(A)+RNA. These results are consistent with the view that some form of translational control involving stored mRNA operates during dark-imbibition and photoinduced germination of spores.

NASA Discipline Number 29-20↗

Germination and elongation of flax in microgravity

This experiment was conducted as part of a risk mitigation payload aboard the Space Shuttle Atlantis on STS-101. The objectives were to test a newly developed water delivery system, and to determine the optimal combination of water volume and substrate for the imbibition and germination of flax (Linum usitatissimum) seeds in space. Two different combinations of germination paper were tested for their ability to absorb, distribute, and retain water in microgravity. A single layer of thick germination paper was compared with one layer of thin germination paper under a layer of thick paper. Paper strips were cut to fit snugly into seed cassettes, and seeds were glued to them with the micropyle ends pointing outward. Water was delivered in small increments that traveled through the paper via capillary action. Three water delivery volumes were tested, with the largest (480 microliters) outperforming the 400 microliters and 320 microliters volumes for percent germination (90.6%) and root growth (mean=4.1 mm) during the 34-hour spaceflight experiment. The ground control experiment yielded similar results, but with lower rates of germination (84.4%) and shorter root lengths (mean=2.8 mm). It is not clear if the roots emerged more quickly in microgravity and/or grew faster than the ground controls. The single layer of thick germination paper generally exhibited better overall growth than the two layered option. Significant seed position effects were observed in both the flight and ground control experiments. Overall, the design of the water delivery system, seed cassettes and the germination paper strip concept was validated as an effective method for promoting seed germination and root growth under microgravity conditions. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

Non-NASA Center↗

Oxygen requirement of germinating flax seeds

Plant experiments in earth orbit are typically prepared on the ground and germinated in orbit to study gravity effects on the developing seedlings. Germination requires the breakdown of storage compounds, and this metabolism depends upon respiration, making oxygen one of the limiting factors in seed germination. In microgravity lack of run-off of excess water requires careful testing of water dispensation and oxygen availability. In preparation for a shuttle experiment (MICRO on STS-107) we studied germination and growth of flax (Linum usitatissimum L.) seedlings in the developed hardware (Magnetic Field Chamber, MFC). We tested between four to 32 seeds per chamber (air volume=14 mL) and after 36 h measured the root length. At 90 microliters O2 per seed (32 seeds/chamber), the germination decreased from 94 to 69%, and the root length was reduced by 20%, compared to 8 seeds per chamber. Based on the percent germination and root length obtained in controlled gas mixtures between 3.6 and 21.6% O2 we determined the lower limit of reliable germination to be 10 vol. % O2 at atmospheric pressure. Although the oxygen available in the MFC's can support the intended number of seeds, the data show that seed storage and microgravity-related limitations may reduce germination. c2003 Published by Elsevier Ltd on behalf of COSPAR.

NASA Discipline Plant Biology↗

Mixed-Linkage Glucan Is the Main Carbohydrate Source and Starch Is an Alternative Source during Brachypodium Grain Germination

Seeds of the model grass Brachypodium distachyon are unusual because they contain very little starch and high levels of mixed-linkage glucan (MLG) accumulated in thick cell walls. It was suggested that MLG might supplement starch as a storage carbohydrate and may be mobilised during germination. In this work, we observed massive degradation of MLG during germination in both endosperm and nucellar epidermis. The enzymes responsible for the MLG degradation were identified in germinated grains and characterized using heterologous expression. By using mutants targeting MLG biosynthesis genes, we showed that the expression level of genes coding for MLG and starch-degrading enzymes was modified in the germinated grains of knocked-out cslf6 mutants depleted in MLG but with higher starch content. Our results suggest a substrate-dependent regulation of the storage sugars during germination. These overall results demonstrated the function of MLG as the main carbohydrate source during germination of Brachypodium grain. More astonishingly, cslf6 Brachypodium mutants are able to adapt their metabolism to the lack of MLG by modifying the energy source for germination and the expression of genes dedicated for its use.

59 BASIC BIOLOGICAL SCIENCES↗

Limited period of graviresponsiveness in germinating spores of Ceratopteris richardii

Rhizoids of the fern Ceratopteris richardii Brogn. usually emerge 40 h after germination is initiated by light, and more than 90% of them emerge growing in a downward direction. However, when the spores are germinated on a clinostat, the emerging rhizoids show no preferential orientation. This indicates that under normal 1 g conditions the initial growth direction of rhizoids can be oriented by gravity. If the orientation of the spores is changed 3 h or less after the start of germination, the growth direction of most emerging rhizoids becomes downward relative to the new orientation. However, if the orientation of the spores is changed by 180 degrees 8 h or more after germination is initiated by light, most rhizoids emerge growing upward; i.e., the same direction as if there had been no orientation change. Emerged rhizoids also do not change their direction of growth if their orientation is changed. These results indicate that the growth direction of emerging rhizoids is set by gravity prior to actual emergence, and that the time of full orientation responsiveness is limited to a period ranging from the initiation of germination to about 3-4 h after the start of germination. There is a gravity-oriented nuclear movement beginning at about 13 h after germination, and this movement appears to predict the initial growth direction of rhizoids.

NASA Discipline Plant Biology↗

A Stochastic Intracellular Model of Anthrax Infection With Spore Germination Heterogeneity

We present a stochastic mathematical model of the intracellular infection dynamics of Bacillus anthracis in macrophages. Following inhalation of B. anthracis spores, these are ingested by alveolar phagocytes. Ingested spores then begin to germinate and divide intracellularly. This can lead to the eventual death of the host cell and the extracellular release of bacterial progeny. Some macrophages successfully eliminate the intracellular bacteria and will recover. Here, a stochastic birth-and-death process with catastrophe is proposed, which includes the mechanism of spore germination and maturation of B. anthracis . The resulting model is used to explore the potential for heterogeneity in the spore germination rate, with the consideration of two extreme cases for the rate distribution: continuous Gaussian and discrete Bernoulli. We make use of approximate Bayesian computation to calibrate our model using experimental measurements from in vitro infection of murine peritoneal macrophages with spores of the Sterne 34F2 strain of B. anthracis . The calibrated stochastic model allows us to compute the probability of rupture, mean time to rupture, and rupture size distribution, of a macrophage that has been infected with one spore. We also obtain the mean spore and bacterial loads over time for a population of cells, each assumed to be initially infected with a single spore. Our results support the existence of significant heterogeneity in the germination rate, with a subset of spores expected to germinate much later than the majority. Furthermore, in agreement with experimental evidence, our results suggest that most of the spores taken up by macrophages are likely to be eliminated by the host cell, but a few germinated spores may survive phagocytosis and lead to the death of the infected cell. Finally, we discuss how this stochastic modelling approach, together with dose-response data, allows us to quantify and predict individual infection risk following exposure.

59 BASIC BIOLOGICAL SCIENCES↗

Effects of germination time on phenolics, antioxidant capacity, in vitro phenolic bioaccessibility and starch digestibility in sorghum

Summary The effects of germination time on phenolic, flavonoid, anthocyanin, antioxidant capacities, inhibition capacity of α‐amylase, bioaccessibility of phenolic and digestibility of starch in sorghum were evaluated in this study. The levels of total phenolic, flavonoid and anthocyanin in germinated sorghum for 48 h increased by 39.74%, 37.28% and 52.21%, respectively. Germination also increased the composition of phenolic, flavonoid and anthocyanin in sorghum, and their antioxidant capacity and inhibitory rates of α‐amylase. Additionally, in vitro digestion results showed that phenolic bioaccessibility increased by 10.18%, and digestibility of starch and expected glycaemic index (eGI) decreased by 13.87% and 5.26 in germinated sorghum for 48 h compared with the ungerminated sorghum. These results indicate that germination might be a green method to improve the nutritional quality of sorghum and promote the development of germinated whole‐grain food.

Li, Ren↗

Relationships Between Seed Weight, Germination Potential and Biochemical Reserves of Maritime Pine in Morocco: Elements for Tree Seedlings Improvement

Selection of quality seeds in breeding programs can significantly improve seedling productivity. Germination and biochemical analyses on seeds from ten natural populations of maritime pine (Pinus pinaster Ait.) in Morocco reveals significant differences among populations in seed weight, germination characters and protein content in both dry seeds and megagametophytes. During germination, the mobilization of protein content in megagametophyte is significantly different among populations than sugar content. A strong positive correlation between the germination capacity and the protein content in both dry seeds and megagametophytes indicates that the best populations in term of germination capacity may also be the richest in protein content. The present study finds that seed weight is not a good indicator for quality seed selection, nor is it recommended to increase the degree of germinability. Our results suggest that the pine population in southern Morocco might have adapted to drought conditions as it is characterized by heavy seed weight and lower speed of protein content mobilization in megagametophyte compared to northern populations growing in temperate climate.

Wahid, Nadya↗

Early quantitative method for measuring germination in non-green spores of Dryopteris paleacea using an epifluorescence-microscope technique

A method is described to determine germination by blue-light excited red fluorescence in the positively photoblastic spores of Dryopteris paleacea Sw. This fluorescence is due to chlorophyll as evidenced from 1) a fluorescence-emission spectrum in vivo, where a bright fluorescence around 675 nm is obtained only in red light (R)-irradiated spores and 2) in vitro measurements with acetone extracts prepared from homogenized spores. Significant amounts of chlorophyll can be found only in R-treated spores; this chlorophyll exhibits an emission band around 668 nm, when irradiated with 430 nm light at 21 degrees C. Compared to other criteria for germination, such as swelling of the cell, coat splitting, greening, and rhizoid formation, which require longer periods after induction for their expression, chlorophyll fluorescence can be used to quantify germination after two days. This result is confirmed by fluence-response curves for R-induced spore germination; the same relationship between applied R and germination is obtained by the evaluation with the epifluorescence method 2 days after the light treatment as compared with the evaluation with bright-field microscopy 5 days after the inducing R. Using this technique we show for the first time that Ca2+ contributes to the signal-transduction chain in phytochrome-mediated chlorophyll synthesis in spores of Dryopteris paleacea.

NASA Discipline Plant Biology↗

Lunar Plant Experiment: Testing Plant Germination in Lunar Gravity and Ionizing Radiation

We present an open design for a first plant growth module on the Moon (LPX). The primary science goal of lunar habitat is to investigate germination and initial plant growth when subject to the combined effects of lunar gravity and lunar surface radiation. The LPX module has been designed to be a flexible base unit that can be adapted to fly to the lunar surface on a variety of landers and rovers. LPX has size and shape of a 1 Unit CubeSat (10 centimeters on a side) and a total mass of 0.3 kilograms. The base will contain a seed module holding around 50 Arabidopsis seeds and provide approximately 0.5 liters of normal air at standard pressure. By the action of a small pump, water will be released after landing to initiate germination. Images will document the germination, initial growth, phototropism, and circumnutation of the small plants while sunlight remains available. A CO2 sensor tracks the release of CO2 during germination and the subsequent uptake of CO2 by photosynthesis. Simulations indicate that the CO2 peak level will be an increase by 2250 parts per million after 2 earth-days and then decline to about 1000 parts per million after 7 earth-days. The camera and CO2 sensor interfaces are USB. The water pump initiation requires a 5-volt signal. After initiation of germination by the addition of water, the habitat must be maintained above 22 degrees Centigrade to allow for plant growth and below 27 degrees Centigrade to prevent damage to the plants. This relatively tight temperature tolerance will require thermal control systems using reflecting surfaces and insulation that must be specifically designed for each lander or rover. The plant growth unit will function to tilt angles of 20 degrees along any axis. Total image data volume required to gauge the plant growth rate and leaf area is 1 megabyte (minimum data required) to 40 megabytes (preferred data). The lid of the habitat will allow adequate sunlight for plant growth. To provide for optimum growth, light levels in the plant growth stage need to be between 75 and 150 micromoles per meter squared per second interval.

first plant growth↗

Conceptual Model of Autonomous Seed Germination Habitat for Mars Mission

As human space exploration extends to Mars, the ability to germinate seeds in extraterrestrial environments is becoming a necessity. Recent technological feats such as the development of the European Modular Cultivation System (EMCS) have made botany experiments possible on the International Space Station (ISS). Despite preliminary designs, a biocompatible plant life support system capable of traveling to Mars has yet to be developed. This study focuses on two preparatory measures regarding seed germination in spaceflight: analysis of seed dormancy protocols and compact autonomous habitat development.The objective of this project is to conceptualize a habitat capable of preserving arabidopsis plant seeds on a long duration spaceflight for the purpose of germinating the first plants on Mars. The proposed container will require a compact, low wattage system to provide gas ventilation, artificial light, and water. A visualization system will also need to be developed in order to monitor seed germination remotely. In order to test the effects of dormancy durations on plant viability, we will conduct a ground study to monitor seed germination in seeds which have been dormant for three, six, nine, and twelve months. We will also compare the effects of different sterilization procedures. The results of this study will be instrumental in developing a viable procedure for transferring the first living plants to Mars.

Peter, Jonah↗

APH Experiments: Germination and Seedling Establishment Methods

Plant experiments during ground studies were conducted in the Advanced Plant Habitat (APH) Science Carrier (SC) for ensuring high germination rates and vigorous seedling establishment. The methods constitute a mixture of environmental and planting factors that control moisture and humidity during early seedling establishment. The APH uses granular media that is fertilized with slow release fertilizer pellets. Seeds were sanitized and their germination was tested prior to use in the SC. Seeds were glued in place with guar gum in different wicking materials. It was found that seedlings establishment was reduced if the chamber was too dry or if salt buildup near the seeds was permitted. Thus, germination was carried in media that was flooded and drained, in a chamber held at 80% relative humidity at a low wind speed of 0.3 m/s. After a week the humidity was lowered to 60% RH and wind speed was increased to 0.6 m/s. The germination of different plant species were found to respond differently to germination wicks or to gauze, as well as fertilization rates. Several methods for growing Arabidopsis, wheat, radish and peppers in the APH SC will be presented

O Monje↗

Anaerobic conditions improve germination of a gibberellic acid deficient rice

Dwarf plants are useful in research because multiple plants can be grown in a small area. Rice (Oryza sativa L.) is especially important since its relatively simple genome has recently been sequenced. We are characterizing a gibberellic acid (GA) mutant of rice (japonica cv 'Shiokari,' line N-71) that is extremely dwarf (20 cm tall). Unfortunately, this GA mutation is associated with poor germination (70%) under aerobic conditions. Neither exogenous GA nor a dormancy-breaking heat treatment improved germination. However, 95% germination was achieved by germinating the seeds anaerobically, either in a pure N2 environment or submerged in unstirred tap water. The anaerobic conditions appear to break a mild post-harvest dormancy in this rice cultivar. Copyright 2002 Crop Science Society of America.

Non-NASA Center↗

Gene activity during germination of spores of the fern, Onoclea sensibilis: RNA and protein synthesis and the role of stored mRNA

Pattern of 3H-uridine incorporation into RNA of spores of Onoclea sensibilis imbibed in complete darkness (non-germinating conditions) and induced to germinate in red light was followed by oligo-dT cellulose chromatography, gel electrophoresis coupled with fluorography and autoradiography. In dark-imbibed spores, RNA synthesis was initiated about 24 h after sowing, with most of the label accumulating in the high mol. wt. poly(A) -RNA fraction. There was no incorporation of the label into poly(A) +RNA until 48 h after sowing. In contrast, photo-induced spores began to synthesize all fractions of RNA within 12 h after sowing and by 24 h, incorporation of 3H-uridine into RNA of irradiated spores was nearly 70-fold higher than that into dark-imbibed spores. Protein synthesis, as monitored by 3H-arginine incorporation into the acid-insoluble fraction and by autoradiography, was initiated in spores within 1-2 h after sowing under both conditions. Autoradiographic experiments also showed that onset of protein synthesis in the cytoplasm of the germinating spore is independent of the transport of newly synthesized nuclear RNA. One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of 35S-methionine-labelled proteins revealed a good correspondence between proteins synthesized in a cell-free translation system directed by poly(A) +RNA of dormant spores and those synthesized in vivo by dark-imbibed and photo-induced spores. These results indicate that stored mRNAs of O. sensibilis spores are functionally competent and provide templates for the synthesis of proteins during dark-imbibition and germination.

NASA Discipline Number 29-20↗

Zinnia Germination and Lunar Soil Amendment

Germination testing was performed to determine the best method for germinating zinnias. This method will be used to attempt to germinate the zinnia seeds produced in space. It was found that seed shape may be critically important in determining whether a seed will germinate or not. The ability of compost and worm castings to remediate lunar regolith simulant for plant growth was tested. It was found that neither treatment effectively improves plant growth in lunar regolith simulant. A potential method of improving lunar regolith simulant by mixing it with arcillite was discovered.

Reese, Laura↗

A prion-like protein regulator of seed germination undergoes hydration-dependent phase separation

Many organisms evolved strategies to survive desiccation. Plant seeds protect dehydrated embryos from various stressors and can lay dormant for millennia. Hydration is the key trigger to initiate germination, but the mechanism by which seeds sense water remains unresolved. Here we identified an uncharacterized Arabidopsis thaliana prion-like protein we named FLOE1, which phase separates upon hydration and allows the embryo to sense water stress. We demonstrate that biophysical states of FLOE1 condensates modulate its biological function in vivo in suppressing seed germination under unfavorable environments. We find intragenic, intraspecific, and interspecific natural variation in FLOE1 expression and phase separation and show that intragenic variation is associated with adaptive germination strategies in natural populations. This combination of molecular, organismal, and ecological studies uncovers FLOE1 as a tunable environmental sensor with direct implications for the design of drought-resistant crops, in the face of climate change.

59 BASIC BIOLOGICAL SCIENCES↗

Switchgrass-associated soil microbes have subtle but distinct effects on germination vs. growth under drought

Soil microbial communities can play a large role in plant fitness, including plants’ response to drought. How microbes mediate drought response may also differ across the plants’ life stage, but studies rarely study causal effects of microbes on plant fitness at multiple stages. We conducted a greenhouse experiment on switchgrass (Panicum virgatum L.) to investigate how microbial presence (sterile bulk vs. live bulk soils) and microbial source (communities from bulk vs. rhizosphere soil) affect germination and seedling growth (life-stage) during drought. We also investigated how drought and life-stage alter the assembly of the inoculated communities. Both drought conditions and the absence of microbes reduced germination (49% and 50% fewer seedlings, respectively, p < 0.05), but there were few effects of microbial presence on older seedlings. In addition, microbial source had no effect on the evaluated plant traits, but interestingly, the initially similar bulk and rhizosphere communities became compositionally distinct after 34 days with germinating seeds, but not with seedlings. Therefore, it may be that the plants did respond to microbial source through changes in seed or root exudates, and calls for further study. Finally, while sometimes microbes did benefit plants under drought, we did not find evidence that they are especially beneficial in this condition. We suggest that microbes’ effects differ across plant life stage, and may be both positive and negative. Further research should advance understanding of plant stress tolerance within a framework of plant-microbiome co-development.

drought↗