Qiuju Liang

Agronomic biofortification of horticultural crops using plant growth-promoting rhizobacteria (PGPR) under crop residue incorporation systems remains largely underexploited. Bacillus subtilis (B1), Bacillus laterosporus (B2), or Bacillus amyloliquefaciens (B3) was inoculated on soil containing chili residue, while chili residue without PGPR (NP) served as the control. Two hybrid long cayenne peppers, succeeding a leaf mustard crop were used in the intensive cultivation study. Net photosynthesis, leaf stomatal conductance, transpiration rate, photosynthetic water use efficiency, shoot and root biomass, and fruit yield were evaluated. Derivatives of folate, minerals, and nitrate contents in the pepper fruits were also assessed. B1 elicited higher net photosynthesis and photosynthetic water use efficiency, while B2 and B3 had higher transpiration rates than B1 and NP. B1 and B3 resulted in 27–36% increase in pepper fruit yield compared to other treatments, whereas B3 produced 24–27.5% and 21.9–27.2% higher 5-methyltetrahydrofolate and total folate contents, respectively, compared to B1 and NP. However, chili residue without PGPR inoculation improved fruit calcium, magnesium, and potassium contents than the inoculated treatments. ‘Xin Xian La 8 F1’ cultivar had higher yield and plant biomass, fruit potassium, total soluble solids, and total folate contents compared to ‘La Gao F1.’ Agronomic biofortification through the synergy of Bacillus amyloliquefaciens and chili residue produced better yield and folate contents with a trade-off in the mineral contents of the greenhouse-grown long cayenne pepper.

Folates are indispensable for plant development, but their molecular mode of action remains elusive. We synthesized a probe, “5-F-THF-Dayne,” comprising 5-formyl-tetrahydrofolate (THF) coupled to a photoaffinity tag. Exploiting this probe in an affinity proteomics study in Arabidopsis thaliana, we retrieved 51 hits. Thirty interactions were independently validated with in vitro expressed proteins to bind 5-F-THF with high or low affinity. Interestingly, the interactors reveal associations beyond one-carbon metabolism, covering also connections to nitrogen (N) metabolism, carbohydrate metabolism/photosynthesis, and proteostasis. Two of the interactions, one with the folate biosynthetic enzyme DIHYDROFOLATE REDUCTASE-THYMIDYLATE SYNTHASE 1 (AtDHFR-TS1) and another with N metabolism-associated glutamine synthetase 1;4 (AtGLN1;4), were further characterized. In silico and experimental analyses revealed G35/K36 and E330 as key residues for the binding of 5-F-THF in AtDHFR-TS1 and AtGLN1;4, respectively. Site-directed mutagenesis of AtGLN1;4 E330, which co-localizes with the ATP-binding pocket, abolished 5-F-THF binding as well as AtGLN1;4 activity. Furthermore, 5-F-THF was noted to competitively inhibit the activities of AtDHFR-TS1 and AtGLN1;4. In summary, we demonstrated a regulatory role for 5-F-THF in N metabolism, revealed 5-F-THF-mediated feedback regulation of folate biosynthesis, and identified a total of 14 previously unknown high-affinity binding cellular targets of 5-F-THF. Together, this sets a landmark toward understanding the role of folates in plant development.

The aim of the present study was to analyze folate content and composition in foods consumed daily by Chinese people. The concentration of seven folate derivatives in sixty‐four selected foods was determined by a liquid‐chromatography tandem‐mass spectrometry method. The total folate levels ranged from 0.28 to 129 µg/100 g fresh weight, with an average of 21.18 μg/100 g. The highest folate content was found in boiled egg yolk and waxy corn (>120 µg/100 g), abundant folate levels in cooked vegetables such as hot pepper, spinach, soybean sprout, stem lettuce, coriander, and broccoli (44–72 µg/100 g), and lowest in Coca Cola (0.28 µg/100 g). 5‐Methyl‐tetrahydrofolate was the major folate derivative in various foods, accounting for 72% of the total folates on average, with the highest being 90% in egg yolk. These data will enable estimation of the daily folate intake and allow dietary recommendations to improve folate status in humans.