Like fungi and some prokaryotes, plants use a thiazole synthase (THI4) to make the thiazole precursor of thiamin. Fungal THI4s are suicide enzymes that destroy an essential active-site Cys residue to obtain the sulfur atom needed for thiazole formation. In contrast, certain prokaryotic THI4s have no active-site Cys, use sulfide as sulfur donor, and are truly catalytic. The presence of a conserved active-site Cys in plant THI4s and other indirect evidence implies that they are suicidal. To confirm this, we complemented the Arabidopsistz-1 mutant, which lacks THI4 activity, with a His-tagged Arabidopsis THI4 construct. LC–MS analysis of tryptic peptides of the THI4 extracted from leaves showed that the active-site Cys was predominantly in desulfurated form, consistent with THI4 having a suicide mechanism in planta. Unexpectedly, transcriptome data mining and deep proteome profiling showed that barley, wheat, and oat have both a widely expressed canonical THI4 with an active-site Cys, and a THI4-like paralog (non-Cys THI4) that has no active-site Cys and is the major type of THI4 in developing grains. Transcriptomic evidence also indicated that barley, wheat, and oat grains synthesize thiamin de novo, implying that their non-Cys THI4s synthesize thiazole. Structure modeling supported this inference, as did demonstration that non-Cys THI4s have significant capacity to complement thiazole auxotrophy in Escherichia coli. There is thus a prima facie case that non-Cys cereal THI4s, like their prokaryotic counterparts, are catalytic thiazole synthases. Bioenergetic calculations show that, relative to suicide THI4s, such enzymes could save substantial energy during the grain-filling period.
Like fungi and some prokaryotes, plants use a thiazole synthase (THI4) to make the thiazole precursor of thiamin. Fungal THI4s are suicide enzymes that destroy an essential active-site Cys residue to obtain the sulfur atom needed for thiazole formation. In contrast, certain prokaryotic THI4s have no active-site Cys, use sulfide as sulfur donor, and are truly catalytic. The presence of a conserved activesite Cys in plant THI4s and other indirect evidence implies that they are suicidal. To confirm this, we complemented the Arabidopsis tz-1 mutant, which lacks THI4 activity, with a His-tagged Arabidopsis THI4 construct. LC-MS analysis of tryptic peptides of the THI4 extracted from leaves showed that the active-site Cys was predominantly in desulfurated form, consistent with THI4 having a suicide mechanism in planta. Unexpectedly, transcriptome datamining and deep proteome profiling showed that barley, wheat, and oat have both a widely expressed canonical THI4 with an active-site Cys, and a THI4-like paralog (non-Cys THI4) that has no active-site Cys and is the major type of THI4 in developing grains. Transcriptomic evidence also indicated that barley, wheat, and oat grains synthesize thiamin de novo, implying that their non-Cys THI4s synthesize thiazole. Structure modeling supported this inference, as did demonstration that non-Cys THI4s have significant capacity to complement thiazole auxotrophy in Escherichia coli. There is thus a prima facie case that non-Cys cereal THI4s, like their prokaryotic counterparts, are catalytic thiazole synthases. Bioenergetic calculations show that, relative to suicide THI4s, such enzymes could save substantial energy during the grain filling period. Short title: Suicidal and catalytic plant THI4 enzymes
A universal lexicon to describe the appearance, aroma/flavors, and textures/feeling factors of peaches was developed. The objective was to provide a standardized lexicon for descriptive validation. A trained descriptive panel established 29 attributes using 51 peach cultivars grown throughout the production season. This lexicon includes 18 aroma and flavor attributes to describe mature peaches as well as under‐ripe and over‐ripe, redness of flesh for appearance, three feeling factors, and seven terms for describing textures. Principal component analysis was used to discern if differences were found among peach samples using the lexicon terms utilized by trained panelists. Texture was the primary differentiating factor in the first dimension of the biplot followed by peach‐identity in the second dimension. Additionally, the attributes “peach‐identity,” “fruity,” “sweet,” “tart,” “citrus,” “sour,” along with textures and feeling factors were prominent in all peach varieties. This lexicon can be useful to identify and quantify sensory attributes in fresh peaches for food and agriculture research. Practical applications The assessment of peach fruit varieties grown throughout the southeastern United States would create a basis for understanding the prominent and unique characteristics of peach varieties and their inherited variability. The peach lexicon created in this study will provide a platform for researchers and producers to understand the desirable sensory traits in peaches. It will allow comparisons among varieties currently available, create a database to be used in breeding applications, and help the growers to produce peaches with desirable sensory traits that could be commercially successful.
Peach production in Georgia, USA, extends from mid-May to mid-August. Multiple cultivars are commercially grown in the U.S., and each cultivar has unique fruit quality characteristics, which could influence consumer perception and acceptability. Among those, peach flavor has been minimally characterized among cultivars. Headspace-solid phase microextraction (HS–SPME) combined with gas chromatography–mass spectrometry (GC–MS) was used to characterize the volatile profile of 42 peach cultivars commercially grown in 2016. The goal of this research was to understand the aroma composition and content of peach cultivars grown in Georgia, USA. Thirty-six volatile compounds were identified. Significant differences (p ≤ 0.0001) were observed for all the flavor volatiles across all cultivars being evaluated. Esters were the major compounds isolated from the 42 peach cultivars, followed by lactones, alcohols, aldehydes, and terpenoids, in that order. Lactones, benzaldehyde, and linalool, which are known to be peach aroma compounds, exhibited the highest mean concentrations among the cultivars with ɣ- and δ-decalactones having a concentration of approximately 180 ng·g−1 and 60 ng·g−1, respectively. Among the cultivars, “Majestic” had the highest mean concentration of ɣ-decalactone, followed by “Southern Pearl” with approximately 503 ng·g−1 and 443 ng·g−1, respectively. “Southern Pearl” and “Fairtime” also exhibited the highest average concentration of δ-decalactone. The results obtained will help provide an understanding of the distribution of the various volatile organic compounds found in Georgia-grown peach cultivars.
The postharvest life of lettuce (Lactuca sativa) is variable and negatively affected by mechanical injury, incomplete cooling, and poor genetic quality. Lettuce breeders are developing cultivars with a longer shelf life and rely on subjective, destructive, and time-consuming methods for quality analysis. One method of accelerating quality evaluations is known as accelerated shelf-life testing (ASLT), which has the potential to assist breeders in assessing lettuce quality and shelf life. The objective of this research was to determine the quality traits that significantly affect shelf life to develop an ASLT procedure to rapidly assess the postharvest quality of lettuce accessions in breeding programs. In Test 1, Romaine lettuce quality was evaluated using one subjective and five objective parameters during storage at 5, 10, 15, or 20 °C. Results determined that weight loss, lightness*, and hue* angle were best correlated with the overall appearance rating, whereas storage at 10 or 15 °C differentiated the shelf-life potential quickly and without excessive deterioration. In Test 2, these objective characteristics and storage temperatures were used to study rates of quality deterioration of a commercial Romaine cultivar (Okeechobee) and a breeding line (60182), both with long shelf lives, and a Batavia lettuce cultivar (La Brillante) with a short shelf life. Lettuce was evaluated during storage at 10 °C (winter and spring seasons) or at 15 °C (winter season). Weight loss was the most appropriate quality index for lettuce at these storage temperatures for a single harvest, whereas lightness* and hue* angle were the most appropriate indices for comparing quality between harvests. To apply ASLT to postharvest assessments of lettuce, breeders and other researchers should include two controls with good and poor shelf life (similar to ‘Okeechobee’ and ‘La Brillante’, respectively) as standard baseline cultivars during storage at either 10 or 15 °C.
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