The protective influence of family support on anxiety, depressive symptoms, and suicidal ideation among elderly Chinese nursing home residents: A study of serial mediation.

In light of the aging demographic in China, heightened attention is warranted for the mental well-being of elderly individuals. Nevertheless, the prevalence of suicidal ideation among older residents in Chinese nursing homes and the nuanced impact of family support on this phenomenon, mediated by anxiety and depressive symptoms, remain unclear. A cohort of 506 Chinese elderly adults participated in the study. Psychosocial traits were assessed using the Perceived Social Support from Family scale (PSS-Fa) for family support, the 7-item Generalized Anxiety Disorder scale (GAD-7) for anxiety symptoms, the 9-item Patient Health Questionnaire (PHQ-9) for depressive symptoms, and suicidal ideation. A structural equation model (SEM) was employed to execute a serial mediation model. The analysis of 506 elderly adults revealed that 8.1% reported varying levels of suicidal ideation within Chinese nursing homes. The pathway from family support to anxiety symptoms (standardized beta = -0.025, P = .241), family support to depressive symptoms (standardized beta = -0.072, P < .05), and family support to suicidal ideation (standardized beta = -0.082, P < .05) were explored. Additionally, pathways from anxiety symptoms to suicidal ideation (standardized beta = 0.364, P < .001), anxiety symptoms to depressive symptoms (standardized beta = 0.647, P < .001), and depressive symptoms to suicidal ideation (standardized beta = 0.369, P < .001) were examined. This study elucidated the underlying mechanisms connecting family support to suicidal ideation, with depressive symptoms partially mediating this association. Additionally, our discoveries shed light on the partial mediation of anxiety symptoms by depressive factors when it came to the realm of suicidal ideation.

CRISPR/Cas9-mediated BoaAOP2s editing alters aliphatic glucosinolate side-chain metabolic flux and increases the glucoraphanin content in Chinese kale.

Glucoraphanin (GRA) is an aliphatic glucosinolate (GSL), and its hydrolysis product has powerful anticancer activity. ALKENYL HYDROXALKYL PRODUCING 2 (AOP2) gene, encodes a 2-oxoglutarate-dependent dioxygenase, which can catalyze GRA to form gluconapin (GNA). However, GRA only present in trace amounts in Chinese kale. To increase the content of GRA in Chinese kale, three copies of BoaAOP2 were isolated and edited using CRISPR/Cas9 system. The content of GRA was 11.71- to 41.29-fold (0.082-0.289 µmol g-1 FW) higher in T1 generation of boaaop2 mutants than in wild-type plants, and this was accompanied by an increase in the GRA/GNA ratio and reductions in the content of GNA and total aliphatic GSLs. BoaAOP2.1 is an effective gene for the alkenylation of aliphatic GSLs in Chinese kale. Overall, targeted editing of CRISPR/Cas9-mediated BoaAOP2s altered aliphatic GSL side-chain metabolic flux and enhanced the GRA content in Chinese kale, suggesting that metabolic engineering of BoaAOP2s has huge potential in improving nutritional quality of Chinese kale.

Sequencing and Analysis of Complete Chloroplast Genomes Provide Insight into the Evolution and Phylogeny of Chinese Kale (Brassica oleracea var. alboglabra).

Chinese kale is a widely cultivated plant in the genus Brassica in the family Brassicaceae. The origin of Brassica has been studied extensively, but the origin of Chinese kale remains unclear. In contrast to Brassica oleracea, which originated in the Mediterranean region, Chinese kale originated in southern China. The chloroplast genome is often used for phylogenetic analysis because of its high conservatism. Fifteen pairs of universal primers were used to amplify the chloroplast genomes of white-flower Chinese kale (Brassica oleracea var. alboglabra cv. Sijicutiao (SJCT)) and yellow-flower Chinese kale (Brassica oleracea var. alboglabra cv. Fuzhouhuanghua (FZHH)) via PCR. The lengths of the chloroplast genomes were 153,365 bp (SJCT) and 153,420 bp (FZHH) and both contained 87 protein-coding genes and eight rRNA genes. There were 36 tRNA genes in SJCT and 35 tRNA genes in FZHH. The chloroplast genomes of both Chinese kale varieties, along with eight other Brassicaceae, were analyzed. Simple sequence repeats, long repeats, and variable regions of DNA barcodes were identified. An analysis of inverted repeat boundaries, relative synonymous codon usage, and synteny revealed high similarity among the ten species, albeit the slight differences that were observed. The Ka/Ks ratios and phylogenetic analysis suggest that Chinese kale is a variant of B. oleracea. The phylogenetic tree shows that both Chinese kale varieties and B. oleracea var. oleracea were clustered in a single group. The results of this study suggest that white and yellow flower Chinese kale comprise a monophyletic group and that their differences in flower color arose late in the process of artificial cultivation. Our results also provide data that will aid future research on genetics, evolution, and germplasm resources of Brassicaceae.

Enhancing health-promoting isothiocyanates in Chinese kale sprouts via manipulating BoESP.

Glucosinolates (GSLs) are a group of sulfur-containing secondary metabolites, which are abundant in Brassica vegetables. GSL breakdown products (GBPs), especially isothiocyanates (ITCs) benefit human health. Chinese kale is a native Brassica vegetable in China, and its sprouts are rich in GSLs and nutritional substances. ITCs are the predominant GBPs while alternative products are formed in the presence of specifier proteins. However, fewer ITCs are formed in the sprouts. Epithiospecifier (ESP) promotes the formation of epithionitriles at the expense of ITCs in Arabidopsis, but a systematic study of different isoforms of ESPs in most vegetables is still missing. In this study, changes in the content of GBPs and the precursor GSLs, as well as thiols per plant were monitored during sprout development. The proportions of epithionitriles and ITCs in total GBPs were found to be increased and decreased, respectively. RNA-seq showed enhanced expression of numerous genes involved in GSLs biosynthesis and degradation, as well as sulfur assimilation in sprouts compared to seeds. Four copies of BoESPs were isolated and BoESP2 was the most abundant isoform. Generally, transcription of BoESPs showed a strong response to abscisic acid and gibberellin, and consequently epithionitriles increased under these treatments. Knockdown of BoESP2 expression through virus-induced gene silencing system could effectively increase total ITCs and decrease total epithionitriles. Overall, dynamic GSL metabolic flux exists in the sprouting period, and the expression of BoESPs determines the pattern of GBPs, suggesting that improving the health-promoting ITCs in Chinese kale sprouts through manipulating BoESPs by metabolic engineering is feasible.

Brassinosteroids fine-tune secondary and primary sulfur metabolism through BZR1-mediated transcriptional regulation.

For adaptation to ever-changing environments, plants have evolved elaborate metabolic systems coupled to a regulatory network for optimal growth and defense. Regulation of plant secondary metabolic pathways such as glucosinolates (GSLs) by defense phytohormones in response to different stresses and nutrient deficiency has been intensively investigated, while how growth-promoting hormone balances plant secondary and primary metabolism has been largely unexplored. Here, we found that growth-promoting hormone brassinosteroid (BR) inhibits GSLs accumulation while enhancing biosynthesis of primary sulfur metabolites, including cysteine (Cys) and glutathione (GSH) both in Arabidopsis and Brassica crops, fine-tuning secondary and primary sulfur metabolism to promote plant growth. Furthermore, we demonstrate that of BRASSINAZOLE RESISTANT 1 (BZR1), the central component of BR signaling, exerts distinct transcriptional inhibition regulation on indolic and aliphatic GSL via direct MYB51 dependent repression of indolic GSL biosynthesis, while exerting partial MYB29 dependent repression of aliphatic GSL biosynthesis. Additionally, BZR1 directly activates the transcription of APR1 and APR2 which encodes rate-limiting enzyme adenosine 5'-phosphosulfate reductases in the primary sulfur metabolic pathway. In summary, our findings indicate that BR inhibits the biosynthesis of GSLs to prioritize sulfur usage for primary metabolites under normal growth conditions. These findings expand our understanding of BR promoting plant growth from a metabolism perspective.

WRKY33-mediated indolic glucosinolate metabolic pathway confers resistance against Alternaria brassicicola in Arabidopsis and Brassica crops.

The tryptophan (Trp)-derived plant secondary metabolites, including camalexin, 4-hydroxy-indole-3-carbonylnitrile, and indolic glucosinolate (IGS), show broad-spectrum antifungal activity. However, the distinct regulations of these metabolic pathways among different plant species in response to fungus infection are rarely studied. In this study, our results revealed that WRKY33 directly regulates IGS biosynthesis, notably the production of 4-methoxyindole-3-ylmethyl glucosinolate (4MI3G), conferring resistance to Alternaria brassicicola, an important pathogen which causes black spot in Brassica crops. WRKY33 directly activates the expression of CYP81F2, IGMT1, and IGMT2 to drive side-chain modification of indole-3-ylmethyl glucosinolate (I3G) to 4MI3G, in both Arabidopsis and Chinese kale (Brassica oleracea var. alboglabra Bailey). However, Chinese kale showed a more severe symptom than Arabidopsis when infected by Alternaria brassicicola. Comparative analyses of the origin and evolution of Trp metabolism indicate that the loss of camalexin biosynthesis in Brassica crops during evolution might attenuate the resistance of crops to Alternaria brassicicola. As a result, the IGS metabolic pathway mediated by WRKY33 becomes essential for Chinese kale to deter Alternaria brassicicola. Our results highlight the differential regulation of Trp-derived camalexin and IGS biosynthetic pathways in plant immunity between Arabidopsis and Brassica crops.

Characterization of the Role of the Neoxanthin Synthase Gene BoaNXS in Carotenoid Biosynthesis in Chinese Kale.

Chinese kale (Brassica oleracea var. alboglabra) is rich in carotenoids, and neoxanthin is one of the most important carotenoids in Chinese kale. In this study, the function of the neoxanthin synthase gene (BoaNXS) in Chinese kale was investigated. BoaNXS, which had a 699-bp coding sequence, was cloned from the white flower cultivar of Chinese kale and was expressed in all developmental stages and organs of Chinese kale; its expression was highest in young seeds. The subcellular localization indicated that BoaNXS was localized in the chloroplast. BoaNXS-overexpressed plants were obtained via Agrobacterium-mediated transient overexpression methodology, and the gene overexpression efficiencies ranged from 2.10- to 4.24-fold. The color in the leaves of BoaNXS-overexpressed plants changed from green to yellow-green; the content of total and individual carotenoids, such as neoxanthin, violaxanthin, and lutein, was significantly increased, and the expression levels of most carotenoid biosynthetic genes were notably increased. These findings indicated that BoaNXS is of vital importance in carotenoid biosynthesis in Chinese kale and could be used as a candidate gene for enriching the carotenoid accumulation and color of Chinese kale and other Brassica vegetables.

Characterization of BoaCRTISO Reveals Its Role in Carotenoid Biosynthesis in Chinese Kale.

Carotenoids are organic pigments that play an important role in both plant coloration and human health; they are a critical subject in molecular breeding due to growing demand for natural molecules in both food and medicine. In this study, we focus upon characterizing BoaCRTISO, the carotenoid isomerase gene before the branch of the carotenoid biosynthetic pathway, which is expressed in all organs and developmental stages of Chinese kale, and BoaCRTISO, which is located in the chloroplast. The expression of BoaCRTISO is induced by strong light, red and blue combined light, and gibberellic acid treatment, but it is suppressed by darkness and abscisic acid treatment. We obtained BoaCRTISO-silenced plants via virus-induced gene silencing technology, and the silence efficiencies ranged from 52 to 77%. The expressions of most carotenoid and chlorophyll biosynthetic genes in BoaCRTISO-silenced plants were downregulated, and the contents of carotenoids and chlorophyll were reduced. Meanwhile, BoaCRTISO-silenced plants exhibited phenotypes of yellowing leaves and inhibited growth. This functional characterization of BoaCRTISO provides insight for the biosynthesis and regulation of carotenoid in Chinese kale.

The flavor of Chinese kale sprouts is affected by genotypic variation of glucosinolates and their breakdown products.

Metabolic profiling of glucosinolates and their breakdown products in sprouts of 22 Chinese kale (Brassica oleracea var. alboglabra, BOA) varieties were investigated by using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). Relationships between glucosinolate metabolites and flavor of Chinese kale sprouts were also analyzed. Results showed that compositions and contents of both glucosinolates and their breakdown products varied greatly among different varieties of Chinese kale sprouts. Gluconapin and 4,5-Epithio-pentanenitrile were the dominant glucosinolate and glucosinolate breakdown product in Chinese kale sprouts, respectively. Gluconapin and glucobrassicin were significantly related to bitterness (r = 0.577, 0.648, respectively; p < 0.05). BOA 1 and BOA 13, BOA 3 and BOA 10 are good candidates for future breeding programs since the former two varieties have light bitterness and pungency, and the latter two varieties contain high levels of glucosinolate breakdown products such as isothiocyanates and epithionitriles in sprouts.

Virus-induced phytohormone dynamics and their effects on plant-insect interactions.

Attacks on plants by both viruses and their vectors is common in nature. Yet the dynamics of the plant-virus-vector tripartite system, in particular the effects of viral infection on plant-insect interactions, have only begun to emerge in the last decade. Viruses can modulate the interactions between insect vectors and plants via the jasmonate, salicylic acid and ethylene phytohormone pathways, resulting in changes in fitness and viral transmission capacity of their insect vectors. Virus infection of plants may also modulate other phytohormones, such as auxin, gibberellins, cytokinins, brassinosteroids and abscisic acid, with yet undefined consequences on plant-insect interactions. Moreover, virus infection in plants may incur changes to other plant traits, such as nutrition and secondary metabolites, that potentially contribute to virus-associated, phytohormone-mediated manipulation of plant-insect interactions. In this article, we review the research progress, discuss issues related to the complexity and variability of the viral modulation of plant interactions with insect vectors, and suggest future directions of research in this field.

Improvement of glucosinolates by metabolic engineering in Brassica crops.

Glucosinolates (GSLs) are a class of sulfur- and nitrogen-containing, and amino acid-derived important secondary metabolites, which mainly present in plants of Brassicaceae family, including Brassica crops, such as broccoli, cabbage, and oilseed rape. The bioactive GSL metabolites confer benefits to plant defense, human health, and the unique flavor of some Brassica crops. However, certain GSL profiles have adverse effects and are known as anti-nutritional factors. This has attracted mounting attempts to increase beneficial GSLs and reduce detrimental ones in the most commonly consumed Brassica crops. We provide a comprehensive overview of metabolic engineering applied in Brassica crops to achieve this purpose, including modulation of GSL biosynthesis, ablation of GSL hydrolysis, inhibition of GSL transport processes, and redirection of metabolic flux to GSL. Moreover, advances in omics approaches, i.e., genomics, transcriptome, and metabolome, applied in the elucidation of GSL metabolism in Brassica crops, as well as promising and potential genome-editing technologies are also discussed.

Effect of melatonin treatment on visual quality and health-promoting properties of broccoli florets under room temperature.

Effect of melatonin treatment on visual quality and contents of health-promoting compounds of broccoli florets under room temperature was investigated in the present study. Broccoli florets were treated with 1 µM melatonin and then stored at room temperature. Results showed that melatonin treatment could delay the post-harvest senescence of broccoli, and performed well in maintaining higher levels of antioxidants, such as carotenoids, vitamin C and total phenols, as well as higher antioxidant capacity than the control. Besides, 1 µM melatonin treatment sustained higher content of glucosinolates, and also resulted in increased percentage of the most potent anticarcinogenic profile, glucoraphanin. Further analysis revealed that 1 µM melatonin strongly induced the expression of glucosinolate biosynthesis-related genes BoMYB28, BoMYB34, BoCYP79F1, and BoCYP79B2, as well as BoTGG1, a gene involved in glucosinolate hydrolysis. In conclusion, post-harvest treatment with 1 µM melatonin is potential in maintaining visual quality and health-promoting properties of broccoli florets.

Combined treatment of epi-brassinolide and NaCl enhances the main phytochemicals in Chinese kale sprouts.

The effects of individual epi-brassinolide (eBL) and NaCl, as well as their combination on contents of main phytochemicals and antioxidant capacity of Chinese kale sprouts were investigated. Our results showed that the application of 100 nM eBL decreased the contents of individual and total glucosinolates, while treatments of 160 mM NaCl both alone and combined with 100 nM eBL enhanced the glucosinolates accumulation by promoting the expression of genes involved in glucosinolate biosynthesis in Chinese kale sprouts and the combined treatment led to significantly higher content of most glucosinolate profiles. Moreover, it also elevated the contents of ascorbic acid and total carotenoids, whereas did not influence the total phenolics and antioxidant capacity. These findings indicated that the combined treatment of 100 nM eBL plus 160 mM NaCl could provide a new strategy to improve the main health promoting compounds in Chinese kale sprouts.

The γ-tubulin complex protein GCP6 is crucial for spindle morphogenesis but not essential for microtubule reorganization in Arabidopsis.

γ-Tubulin typically forms a ring-shaped complex with 5 related γ-tubulin complex proteins (GCP2 to GCP6), and this γ-tubulin ring complex (γTuRC) serves as a template for microtubule (MT) nucleation in plants and animals. While the γTuRC takes part in MT nucleation in most eukaryotes, in fungi such events take place robustly with just the γ-tubulin small complex (γTuSC) assembled by γ-tubulin plus GCP2 and GCP3. To explore whether the γTuRC is the sole functional γ-tubulin complex in plants, we generated 2 mutants of the GCP6 gene encoding the largest subunit of the γTuRC in Arabidopsis thaliana. Both mutants showed similar phenotypes of dwarfed vegetative growth and reduced fertility. The gcp6 mutant assembled the γTuSC, while the wild-type cells had GCP6 join other GCPs to produce the γTuRC. Although the gcp6 cells had greatly diminished γ-tubulin localization on spindle MTs, the protein was still detected there. The gcp6 cells formed spindles that lacked MT convergence and discernable poles; however, they managed to cope with the challenge of MT disorganization and were able to complete mitosis and cytokinesis. Our results reveal that the γTuRC is not the only functional form of the γ-tubulin complex for MT nucleation in plant cells, and that γ-tubulin-dependent, but γTuRC-independent, mechanisms meet the basal need of MT nucleation. Moreover, we show that the γTuRC function is more critical for the assembly of spindle MT array than for the phragmoplast. Thus, our findings provide insight into acentrosomal MT nucleation and organization.

Main Health-Promoting Compounds Response to Long-Term Freezer Storage and Different Thawing Methods in Frozen Broccoli Florets.

The effects of long-term freezer storage and different defrosting methods on the retention of glucosinolates, vitamin C, and total phenols in frozen broccoli florets were investigated in the present study. Frozen broccoli florets were stored in a freezer at -20 °C for 165 days or subjected to defrosting by three different house-hold thawing methods (water, air, and refrigerator defrosting). Results showed that all glucosinolates were well preserved, while vitamin C and total phenols were reduced by less than 12% and 19% of the control, respectively, during the storage. Besides, refrigerator and air defrosting were better than water defrosting in glucosinolates retention, and refrigerator defrosting was the best in vitamin C preservation. No difference was observed in reserving phenolic compounds among the three methods. In conclusion, long-term freezer storage is an excellent way to preserve broccoli florets, and refrigerator defrosting is the best way to maintain the nutritional compounds in frozen broccoli florets.

Functional Characterization of BoaMYB51s as Central Regulators of Indole Glucosinolate Biosynthesis in Brassica oleracea var. alboglabra Bailey.

R2R3-MYB transcription factor MYB51 is known to control indole glucosinolate (indole GSL) biosynthesis in Arabidopsis. Here, two copies of BoaMYB51 have been isolated in Chinese kale (Brassica oleracea var. alboglabra Bailey), designated BoaMYB51.1 and BoaMYB51.2, which exhibit overlapping but distinct expression levels among different organs and respond to signaling molecules in a similar pattern. It has been demonstrated a structural and functional conservation between BoaMYB51s and AtMYB51 by phylogenetic analysis, complementation studies and transient expression assay. To further investigate the transcriptional mechanism, we identified the transcriptional activation domain (TAD) and putative interacting proteins of BoaMYB51s by means of yeast (Saccharomyces cerevisiae) two hybrid. Using tobacco (Nicotiana benthamiana) transient expression assay, we confirmed that the carboxy-end is required for transcriptional activation activity of BoaMYB51s. In addition, several BoaMYB51-interacting proteins have been identified by yeast two-hybrid screening. These results provide important insights into the molecular mechanisms by which MYB51 transcriptionally regulates indole GSL biosynthesis.

Effects of postharvest methyl jasmonate treatment on main health-promoting components and volatile organic compounds in cherry tomato fruits.

Effects of postharvest methyl jasmonate (MeJA) treatment on the contents of ascorbic acid and carotenoids, as well as the compositions and contents of volatile organic compounds (VOCs) in cherry tomato fruits were investigated during 11 days of storage at room temperature (25 °C). The results showed that MeJA treatment significantly increased the contents of ascorbic acid and carotenoids, especially lycopene in postharvest cherry tomato fruits. Moreover, MeJA treatment improved the contents of carotenoids derived VOCs such as 6-methyl-5-hepten-2-one (MHO), while had no effect on firmness, sugars and titratable acidity. All above results suggested that the exogenous MeJA application is potential in enhancement of main health-promoting components and VOCs in postharvest cherry tomato fruits.

Effects of glucose and gibberellic acid on glucosinolate content and antioxidant properties of Chinese kale sprouts.

Glucosinolates, anthocyanins, total phenols, and vitamin C, as well as antioxidant capacity, were investigated in Chinese kale sprouts treated with both glucose and gibberellic acid (GA3). The combination of 3% (0.03 g/ml) glucose and 5 µmol/L GA3 treatment was effective in increasing glucosinolate content while glucose or GA3 treatment alone did not influence significantly almost all individual glucosinolates or total glucosinolates. The total phenolic content and antioxidant activity of Chinese kale sprouts were enhanced by combined treatment with glucose and GA3, which could be useful in improving the main health-promoting compounds and antioxidant activity in Chinese kale sprouts.

Influence of pre-harvest red light irradiation on main phytochemicals and antioxidant activity of Chinese kale sprouts.

The effects of pre-harvest red light irradiation on main healthy phytochemicals as well as antioxidant activity of Chinese kale sprouts during postharvest storage were investigated. 6-day-old sprouts were treated by red light for 24h before harvest and sampled for further analysis of nutritional quality on the first, second and third day after harvest. The results indicated that red light exposure notably postponed the degradation of aliphatic, indole, and total glucosinolates during postharvest storage. The vitamin C level was remarkably higher in red light treated sprouts on the first and second day after harvest when compared with the control. In addition, red light treatment also enhanced the accumulation of total phenolics and maintained higher level of antioxidant activity than the control. All above results suggested that pre-harvest red light treatment might provide a new strategy to maintain the nutritive value of Chinese kale sprouts during postharvest storage.