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All sources NASA NTRS DOE OSTI

DOE OSTI2021

Methyl jasmonate effects on sugarbeet root responses to postharvest dehydration

Background: Sugarbeet (Beta vulgaris L.) roots are stored under conditions that cause roots to dehydrate, which increases postharvest losses. Although exogenous jasmonate applications can reduce drought stress in intact plants, their ability to alleviate the effects of dehydration in postharvest sugarbeet roots or other stored plant products is unknown. Research was conducted to determine whether jasmonate treatment could mitigate physiological responses to dehydration in postharvest sugarbeet roots. Methods: Freshly harvested sugarbeet roots were treated with 10 µM methyl jasmonate (MeJA) or water and stored under dehydrating and non-dehydrating storage conditions. Changes in fresh weight, respiration rate, wound healing, leaf regrowth, and proline metabolism of treated roots were investigated throughout eight weeks in storage. Results: Dehydrating storage conditions increased root weight loss, respiration rate, and proline accumulation and prevented leaf regrowth from the root crown. Under dehydrating conditions, MeJA treatment reduced root respiration rate, but only in severely dehydrated roots. MeJA treatment also hastened wound-healing, but only in the late stages of barrier formation. MeJA treatment did not impact root weight loss or proline accumulation under dehydrating conditions or leaf regrowth under non-dehydrating conditions. Both dehydration and MeJA treatment affected expression of genes involved in proline metabolism. In dehydrated roots, proline dehydrogenase expression declined 340-fold, suggesting that dehydration-induced proline accumulation was governed by reducing proline degradation. MeJA treatment altered proline biosynthetic and catabolic gene expression, with greatest effect in nondehydrated roots. Overall, MeJA treatment alleviated physiological manifestations of dehydration stress in stored roots, although the beneficial effects were small. Postharvest jasmonate applications, therefore, are unlikely to significantly reduce dehydrationrelated storage losses in sugarbeet roots.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2023

Wounding rapidly alters transcription factor expression, hormonal signaling, and phenolic compound metabolism in harvested sugarbeet roots

Injuries sustained by sugarbeet (Beta vulgaris L.) roots during harvest and postharvest operations seriously reduce the yield of white sugar produced from stored roots. Although wound healing is critically important to reduce losses, knowledge of these processes is limited for this crop as well as for roots in other species. To better understand the metabolic signals and changes that occur in wounded roots, dynamic changes in gene expression were determined by RNA sequencing and the activity of products from key genes identified in this analysis were determined in the 0.25 to 24 h following injury. Nearly five thousand differentially expressed genes that contribute to a wide range of cellular and molecular functions were identified in wounded roots. Highly upregulated genes included transcription factor genes, as well as genes involved in ethylene and jasmonic acid (JA) biosynthesis and signaling and phenolic compound biosynthesis and polymerization. Enzyme activities for key genes in ethylene and phenolic compound biosynthesis and polymerization also increased due to wounding. Results indicate that wounding causes a major reallocation of metabolism in sugarbeet taproots. Although both ethylene and JA are likely involved in triggering wound responses, the greater and more sustained upregulation of ethylene biosynthesis and signaling genes relative to those of JA, suggest a preeminence of ethylene signaling in wounded sugarbeet roots. Changes in gene expression and enzymes involved in phenolic compound metabolism additionally indicate that barriers synthesized to seal off wounds, such as suberin or lignin, are initiated within the first 24 h after injury.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2002

Swiss chard: a salad crop for the space program

Salad greens will be among the first crops grown on lunar or planetary space stations. Swiss chard (Beta vulgaris L.) is an important candidate salad crop because it is high yielding and rich in vitamins and minerals. Five Swiss chard cultivars were grown in the greenhouse under two light levels for 13 weeks to compare cumulative yields from weekly harvests, mineral composition, and to evaluate sensory attributes as a salad green. The varieties Large White Ribbed (LWR) and Lucullus (LUC) were the highest yielding in both light regimes. LWR was the shortest of the cultivars requiring the least vertical space. LWR also received the highest sensory ratings of the five cultivars. LWR Swiss chard should be considered as an initial test variety in food production modules.

NASA Discipline Life Support Systems↗

DOE OSTI2022

Newly developed sugarbeet lines with altered postharvest respiration rates differ in transcription factor and glycolytic enzyme expression

Respiration is the principal cause for postharvest sucrose loss in sugarbeet (Beta vulgaris L.) roots. Although reductions in respiration rate could mitigate these losses, developing sugarbeet cultivars and storage procedures that reduce respiration are hindered by a lack of knowledge of the genetic and metabolic factors that control storage respiration rate. Research was conducted to identify genes and gene products that affect storage respiration rate by creating two sugarbeet lines that differ in respiration rate and characterizing gene expression differences between these lines. Sugarbeet lines F1056 and F1057, which differ by up to 42% in respiration rate, were created by divergent selection of a sugarbeet population using root respiration rate after 30 d in storage as the principal selection criterion. RNA sequencing identified 287 differentially expressed genes (DEGs) between these lines on the day of harvest and after 28-d storage. Of these DEGs, nine encoded transcription factors and five encoded enzymes involved in the respiratory pathway. Other DEGs contributed to a variety of biological and molecular functions based on gene ontology classifications. Of respiratory pathway DEGS, genes for NAD+- and NADP+-dependent forms of glyceraldehyde-3-phosphate dehydrogenase were of note due to their high upregulation in the high respiring line and for their established role in glycolysis, a pathway identified as a likely bottleneck in respiratory substrate production. Overall, lines F1056 and F1057 provide new tools for investigating genetic and physiological differences in storage respiration rate, and their DEGs identify candidates for genes affecting sugarbeet root respiration rate.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2000

Low potassium enhances sodium uptake in red-beet under moderate saline conditions

Due to the discrepancy in metabolic sodium (Na) requirements between plants and animals, cycling of Na between humans and plants is limited and critical to the proper functioning of bio-regenerative life support systems, being considered for long-term human habitats in space (e.g., Martian bases). This study was conducted to determine the effects of limited potassium (K) on growth, Na uptake, photosynthesis, ionic partitioning, and water relations of red-beet (Beta vulgaris L. ssp. vulgaris) under moderate Na-saline conditions. Two cultivars, Klein Bol, and Ruby Queen were grown for 42 days in a growth chamber using a re-circulating nutrient film technique where the supplied K levels were 5.0, 1.25, 0.25, and 0.10 mM in a modified half-strength Hoagland solution salinized with 50 mM NaCl. Reducing K levels from 5.0 to 0.10 mM quadrupled the Na uptake, and lamina Na levels reached -20 g kg-1 dwt. Lamina K levels decreased from -60 g kg-1 dwt at 5.0 mM K to -4.0 g kg-1 dwt at 0.10 mM K. Ruby Queen and Klein Bol responded differently to these changes in Na and K status. Klein Bol showed a linear decline in dry matter production with a decrease in available K, whereas for cv. Ruby Queen, growth was stimulated at 1.25 mM K and relatively insensitive to a further decreases of K down to 0.10 mM. Leaf glycinebetaine levels showed no significant response to the changing K treatments. Leaf relative water content and osmotic potential were significantly higher for both cultivars at low-K treatments. Leaf chlorophyll levels were significantly decreased at low-K treatments, but leaf photosynthetic rates showed no significant difference. No substantial changes were observed in the total cation concentration of plant tissues despite major shifts in the relative Na and K uptake at various K levels. Sodium accounted for 90% of the total cation uptake at the low K levels, and thus Na was likely replacing K in osmotic functions without negatively affecting the plant water status, or growth. Our results also suggest that cv. Ruby Queen can tolerate a much higher Na tissue concentration than cv. Klein Bol before there is any growth reduction. Grant numbers: 12180.

NASA Center KSC↗

NASA NTRS1999

How far can sodium substitute for potassium in red beet?

Sodium (Na) movement between plants and humans is one of the more critical aspects of bioregenerative systems of life support, which NASA is studying for the establishment of long-term bases on the Lunar or Martian surface. This study was conducted to determine the extent to which Na can replace potassium (K) in red beet (Beta vulgaris L. ssp vulgaris) without adversely affecting metabolic functions such as water relations, photosynthetic rates, and thus growth. Two cultivars, Ruby Queen and Klein Bol, were grown for 42 days at 1200 micromoles mol-1 CO2 in a growth chamber using a re-circulating nutrient film technique with 0%, 75%, 95%, and 98% Na substitution for K in a modified half-strength Hoagland solution. Total biomass of Ruby Queen was greatest at 95% Na substitution and equal at 0% and 98% Na substitution. For Klein Bol, there was a 75% reduction in total biomass at 98% Na substitution. Nearly 95% of the total plant K was replaced with Na at 98% Na substitution in both cultivars. Potassium concentrations in leaves decreased from 120 g kg-1 dwt in 0% Na substitution to 3.5 g kg-1 dwt at 98% Na substitution. Leaf chlorophyll concentration, photosynthetic rate, and osmotic potential were not affected in either cultivar by Na substitution for K. Leaf glycinebetaine levels were doubled at 75% Na substitution in Klein Bol, but decreased at higher levels of Na substitution. For Ruby Queen, glycinebetaine levels in leaf increased with the first increase of Na levels and were maintained at the higher Na levels. These results indicate that in some cultivars of red beet, 95% of the normal tissue K can be replaced by Na without a reduction in growth.

NASA Discipline Life Support Systems↗

NASA NTRS1992

Antipeptide antibodies that can distinguish specific subunit polypeptides of glutamine synthetase from bean (Phaseolus vulgaris L.)

The amino acid sequences of the beta and gamma subunit polypeptides of glutamine synthetase from bean (Phaseolus vulgaris L.) root nodules are very similar. However, there are small regions within the sequences that are significantly different between the two polypeptides. The sequences between amino acids 2 and 9 and between 264 and 274 are examples. Three peptides (gamma 2-9, gamma 264-274, and beta 264-274) corresponding to these sequences were synthesized. Antibodies against these peptides were raised in rabbits and purified with corresponding peptide-Sepharose affinity chromatography. Western blot analysis of polyacrylamide gel electrophoresis of bean nodule proteins demonstrated that the anti-beta 264-274 antibodies reacted specifically with the beta polypeptide and the anti-gamma 264-274 and anti-gamma 2-9 antibodies reacted specifically with the gamma polypeptide of the native and denatured glutamine synthetase. These results showed the feasibility of using synthetic peptides in developing antibodies that are capable of distinguishing proteins with similar primary structures.

NASA Program NSCORT↗

DOE OSTI2024

Transcriptomic and metabolomic changes in postharvest sugarbeet roots reveal widespread metabolic changes in storage and identify genes potentially responsible for respiratory sucrose loss

Endogenous metabolism is primarily responsible for losses in sucrose content and processing quality in postharvest sugarbeet roots. The genes responsible for this metabolism and the transcriptional changes that regulate it, however, are largely unknown. To identify genes and metabolic pathways that participate in postharvest sugarbeet root metabolism and the transcriptional changes that contribute to their regulation, transcriptomic and metabolomic profiles were generated for sugarbeet roots at harvest and after 12, 40 and 120 d storage at 5 and 12°C and gene expression and metabolite concentration changes related to storage duration or temperature were identified. During storage, 8656 genes, or 34% of all expressed genes, and 225 metabolites, equivalent to 59% of detected metabolites, were altered in expression or concentration, indicating extensive transcriptional and metabolic changes in stored roots. These genes and metabolites contributed to a wide range of cellular and molecular functions, with carbohydrate metabolism being the function to which the greatest number of genes and metabolites classified. Because respiration has a central role in postharvest metabolism and is largely responsible for sucrose loss in sugarbeet roots, genes and metabolites involved in and correlated to respiration were identified. Seventy-five genes participating in respiration were differentially expressed during storage, including two bidirectional sugar transporter SWEET17 genes that highly correlated with respiration rate. Weighted gene co-expression network analysis identified 1896 additional genes that positively correlated with respiration rate and predicted a pyruvate kinase gene to be a central regulator or biomarker for respiration rate. Overall, these results reveal the extensive and diverse physiological and metabolic changes that occur in stored sugarbeet roots and identify genes with potential roles as regulators or biomarkers for respiratory sucrose loss.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1995

Chemical Topography of Efferent Projections from the Median Preoptic Nucleus to Pontine Monoaminergic Cell Groups in the Rat

This study examined efferent output from the median preoptic nucleus (MnPO) to pontine noradrenergic and serotonergic cell groups using an anterograde tracing technique (Phaseolus vulgaris leucoagglutinin, PHA-L) combined with glucose oxidase immunocytochemistry to scrotonin (5-HT) or to dopamine-(beta)-hydroxylase (DBH). Injections of PHA-L into the ventral MNPO resulted in moderate axonal labeling within the region of the B7 and B8 serotonergic groups in the dorsal raphe. PHA-L labeled fibers and punctate processes were observed in close apposition to many of the 5-HT immunoreactive neurons in these regions, In contrast, sparse terminal labeling was found within the B5 group in the raphe pontis nucleus, and only trace fiber labeling observed in the B3 and B6 groups. Efferents from the MNPO also provided moderate innervation to the A6 and A7 noradrenergic groups. PHA-L labeled punctate processes were found most frequently in close apposition to DBH-immunorcactive neurons at mid- to caudal levels of the locus coeruleus. Some labeled axons were also present within the A7 and A5 groups. Additionally, a close apposition between labeled MNPO efferents and 5-HT fibers within the lateral parabrachial nucleus was observed, The results indicate the MNPO provides a topographic innerva- tion of monoaminergic groups in the upper brainstem.

Zardetto-Smith, Andrea M.↗

NASA NTRS1995

Chemical Topography of Efferent Projections from the Median Preoptic Nucleus to Pontine Monoaminergic Cell Groups in the Rat

This study examined efferent output from the median preoptic nucleus (MNPO) to pontine noradrenergic and serotonergic cell groups using an anterograde tracing technique (Phaseolus vulgaris leucoagglutinin, PHA-L) combined with glucose oxidase immunocytochemistry to serotonin (5-HT) or to dopamine-beta-hydroxylase (DBH). Injections of PHA-L into the ventral MNPO resulted in moderate axonal labeling within the region of the B7 and B8 serotonergic groups in the dorsal raphe. PHA-L labeled fibers and punctate processes were observed in close apposition to many of the 5-HT immunoreactive neurons in these regions. In contrast, sparse terminal labeling was found within the B5 group in the raphe pontis nucleus, and only trace fiber labeling observed in the B3 and B6 groups. Efferents from the MNPO also provided moderate innervation to the A6 and A7 noradrenergic groups. PHA-L labeled punctate processes were found most frequently in close apposition to DBH-immunoreactive neurons at mid- to caudal levels of the locus coeruleus. Some labeled axons were also present within the A7 and A5 groups. Additionally, a close apposition between labeled MNPO efferents and 5-HT fibers within the lateral parabrachial nucleus was observed. The results indicate the MNPO provides a topographic innervation of monoaminergic groups in the upper brainstem.

Zardetto-Smith, Andrea M.↗