Transcription Cofactor CsMBF1c Enhances Heat Tolerance of Cucumber and Interacts with Heat-Related Proteins CsNFYA1 and CsDREB2.

Multiprotein bridging factor 1 (MBF1) is a very important transcription factor (TF) in plants, whose members influence numerous defense responses. Our study found that MBF1c in Cucurbitaceae was highly conserved. CsMBF1c expression was induced by temperature, salt stress, and abscisic acid (ABA) in cucumber. Overexpressed CsMBF1c enhanced the heat resistance of a cucumber, and the Csmbf1c mutant showed decreased resistance to high temperatures (HTs). CsMBF1c played an important role in stabilizing the photosynthetic system of cucumber under HT, and its expression was significantly associated with heat-related TFs and genes related to protein processing in the endoplasmic reticulum (ER). Protein interaction showed that CsMBF1c interacted with dehydration-responsive element binding protein 2 (CsDREB2) and nuclear factor Y A1 (CsNFYA1). Overexpression of CsNFYA1 in Arabidopsis improved the heat resistance. Transcriptional activation of CsNFYA1 was elevated by CsMBF1c. Therefore, CsMBF1c plays an important regulatory role in cucumber's resistance to high temperatures.

Positive effects of extracellular polysaccharides from Paecilomyces hepiali on immune-enhancing properties by regulating gut microbiota in cyclophosphamide-induced mice.

Paecilomyces hepiali is a precious health-care edible medicinal fungus with rich polysaccharides and exhibits various biological activities. Polysaccharides from P. hepiali fermentation broth (PHP) exhibits good immunomodulatory activity; however, the mechanism underlying PHP-mediated regulation of immunity and gut microbiota remains unclear. To reveal the mechanisms, PHP of different doses were used to intervene cyclophosphamide (CTX)-induced immunosuppressive model mice. The results revealed that PHP facilitated the secretion of serum cytokines, increased the mRNA and protein expression of TLR4/NF-κB signaling pathway. Furthermore, it improved the physical barrier function of the intestine by upregulating the expression of tight junction proteins. PHP increased the proliferation of beneficial bacteria, including, Actinobacteriota, Alistipes, Candidatus_Saccharimonas and unclassified_Clostridia_vadinBB60_group, and reduced the abundance of Proteobacteria, Deferribacterota, Mucispirillum and Escherichia_Shigella, promoted the production of short-chain fatty acids, which were positively associated with immune traits. Thus, as an immune enhancer, PHP has the potential to regulate the intestinal immune response in immunosuppressed mice through modulating gut microbiota.

A quick and effective method for thermostability differentiation in cucumber (Cucumis sativus L.).

High temperature affects the growth and production of cucumber. Selecting thermotolerant cucumber cultivars is conducive to coping with high temperatures and improving production. Thus, a quick and effective method for screening thermotolerant cucumber cultivars is needed. In this study, four cucumber cultivars were used to identify heat resistance indexes. The morphological, physiological and biochemical indexes were measured. When exposed to high temperatures, thermotolerant cucumber had a more stable photosystem, membrane, and oxidation-reduction systems. The impact of high temperatures on plants is multifaceted, and the accurate discrimination of heat resistance cannot be achieved solely based on a single or multiple indicators. Therefore, principal component analysis (PCA) was employed to comprehensively evaluate the heat resistance of cucumber plants. The results showed that the heat resistance obtained by PCA was significantly correlated with the heat injury index. In addition, the stepwise regression equation identified two heat-related indices, hydrogen peroxide content (H2 O2 ) and photosynthetic operating efficiency (Fq'/Fm'), and they can quickly distinguish the heat resistance of the other 8 cucumber cultivars. These results will help to accelerate the selection of thermotolerant resources and assist in cucumber breeding.

Modulating effects of Hericium erinaceus polysaccharides on the immune response by regulating gut microbiota in cyclophosphamide-treated mice.

BACKGROUND: The gut microbiota (GM) is recognized as a significant contributor to the immune system. In the present study, the effects of Hericium erinaceus polysaccharides (HEP) on immunoregulation and GM in cyclophosphamide (CTX)-treated mice were investigated to elucidate the attenuate of immunosuppression by modulating GM. RESULTS: The results revealed that HEP significantly improved the body weight and immune organ index in immunodeficient mice (P < 0.05). They significantly increased operational taxonomic units (OTUs) (P < 0.05), adjusted the α and ß diversity of the GM, and the bacterial community structure was more similar to that of control group. Taxonomic composition analysis found that HEP increased the abundance of Alistipse, uncultured_bacterium_f_Muribaculaceae, Lachnospiraceae_NK4A136_group, uncultured_bacterium_f_Lachnospiracea, uncultured_bacterium_f_Ruminococcaceae and Ruminococcaceae_UCG-014, and decreased Lactobacillus, Bacteroides, and Alloprevotella, suggesting that HEP can improve the GM structure and inhibit CTX-induced GM dysregulation. Moreover, HEP increased short-chain fatty acid (SCFA)-producing bacteria, recovered SCFA levels, alleviated immunosuppression caused by CTX, enhanced the serum immune cytokine factors, and upregulated TLR4/NF-κB pathway key proteins (TLR4, NF-κB p65) at mRNA and protein levels. CONCLUSION: Hericium erinaceus polysaccharides effectively regulated GM and enhancement of intestinal immune function, so they have the potential to be developed as functional ingredients or foods to modulate immune responses. © 2022 Society of Chemical Industry.

Systematic analysis of the UDP-glucosyltransferase family: discovery of a member involved in rutin biosynthesis in Solanum melongena.

Eggplant (Solanum melongena) is an economically important crop and rich in various nutrients, among which rutin that has positive effects on human health is found in eggplant. Glycosylation mediated by UDP-glycosyltransferases (UGTs) is a key step in rutin biosynthesis. However, the UGT gene has not been reported in eggplant to date. Herein, 195 putative UGT genes were identified in eggplant by genome-wide analysis, and they were divided into 17 subgroups (Group A-P and Group R) according to the phylogenetic evolutionary tree. The members of Groups A, B, D, E and L were related to flavonol biosynthesis, and rutin was the typical flavonol. The expression profile showed that the transcriptional levels of SmUGT genes in Clusters 7-10 were closely related to those of rutin biosynthetic pathway genes. Notably, SmUGT89B2 was classified into Cluster 7 and Group B; its expression was consistent with rutin accumulation in different tissues and different leaf stages of eggplant. SmUGT89B2 was located in the nucleus and cell membrane. Virus-induced gene silencing (VIGS) and transient overexpression assays showed that SmUGT89B2 can promote rutin accumulation in eggplant. These findings provide new insights into the UGT genes in eggplant, indicating that SmUGT89B2 is likely to encode the final enzyme in rutin biosynthesis.

Genome-Wide Analysis and Characterization of Eggplant F-Box Gene Superfamily: Gene Evolution and Expression Analysis under Stress.

F-box genes play an important role in plant growth and resistance to abiotic and biotic stresses. To date, systematic analysis of F-box genes and functional annotation in eggplant (Solanum melongena) is still limited. Here, we identified 389 F-box candidate genes in eggplant. The domain study of F-box candidate genes showed that the F-box domain is conserved, whereas the C-terminal domain is diverse. There are 376 SmFBX candidate genes distributed on 12 chromosomes. A collinearity analysis within the eggplant genome suggested that tandem duplication is the dominant form of F-box gene replication in eggplant. The collinearity analysis between eggplant and the three other species (Arabidopsis thaliana, rice and tomato) provides insight into the evolutionary characteristics of F-box candidate genes. In addition, we analyzed the expression of SmFBX candidate genes in different tissues under high temperature and bacterial wilt stress. The results identified several F-box candidate genes that potentially participate in eggplant heat tolerance and bacterial wilt resistance. Moreover, the yeast two-hybrid assay showed that several representative F-box candidate proteins interacted with representative Skp1 proteins. Overexpression of SmFBX131 and SmFBX230 in tobacco increased resistance to bacterial wilt. Overall, these results provide critical insights into the functional analysis of the F-box gene superfamily in eggplant and provide potentially valuable targets for heat and bacterial resistance.

Genome-wide identification of the Capsicum bHLH transcription factor family: discovery of a candidate regulator involved in the regulation of species-specific bioactive metabolites.

BACKGROUND: The basic helix-loop-helix (bHLH) transcription factors (TFs) serve crucial roles in regulating plant growth and development and typically participate in biological processes by interacting with other TFs. Capsorubin and capsaicinoids are found only in Capsicum, which has high nutritional and economic value. However, whether bHLH family genes regulate capsorubin and capsaicinoid biosynthesis and participate in these processes by interacting with other TFs remains unknown. RESULTS: In this study, a total of 107 CabHLHs were identified from the Capsicum annuum genome. Phylogenetic tree analysis revealed that these CabHLH proteins were classified into 15 groups by comparing the CabHLH proteins with Arabidopsis thaliana bHLH proteins. The analysis showed that the expression profiles of CabHLH009, CabHLH032, CabHLH048, CabHLH095 and CabHLH100 found in clusters C1, C2, and C3 were similar to the profile of carotenoid biosynthesis in pericarp, including zeaxanthin, lutein and capsorubin, whereas the expression profiles of CabHLH007, CabHLH009, CabHLH026, CabHLH063 and CabHLH086 found in clusters L5, L6 and L9 were consistent with the profile of capsaicinoid accumulation in the placenta. Moreover, CabHLH007, CabHLH009, CabHLH026 and CabHLH086 also might be involved in temperature-mediated capsaicinoid biosynthesis. Yeast two-hybrid (Y2H) assays demonstrated that CabHLH007, CabHLH009, CabHLH026, CabHLH063 and CabHLH086 could interact with MYB31, a master regulator of capsaicinoid biosynthesis. CONCLUSIONS: The comprehensive and systematic analysis of CabHLH TFs provides useful information that contributes to further investigation of CabHLHs in carotenoid and capsaicinoid biosynthesis.

Genome-wide identification of the tea plant bHLH transcription factor family and discovery of candidate regulators of trichome formation.

Leaf trichomes play vital roles in plant resistance and the quality of tea. Basic helix-loop-helix (bHLH) transcription factors (TFs) play an important role in regulating plant development and growth. In this study, a total of 134 CsbHLH proteins were identified in the Camellia sinensis var. sinensis (CSS) genome. They were divided into 17 subgroups according to the Arabidopsis thaliana classification. Phylogenetic tree analysis indicated that members of subgroups IIIc-I and IIIc-II might be associated with trichome formation. The expression patterns of CsbHLH116, CsbHLH133, CsbHLH060, CsbHLH028, CsbHLH024, CsbHLH112 and CsbHLH053 from clusters 1, 3 and 5 were similar to the trichome distribution in tea plants. CsbHLH024 and CsbHLH133 were located in the cell nucleus and possessed transcriptional activation ability. They could interact with CsTTG1, which is a regulator of tea trichome formation. This study provides useful information for further research on the function of CsbHLHs in trichome formation.

The draft genome of Tibetan hulless barley reveals adaptive patterns to the high stressful Tibetan Plateau.

The Tibetan hulless barley (Hordeum vulgare L. var. nudum), also called "Qingke" in Chinese and "Ne" in Tibetan, is the staple food for Tibetans and an important livestock feed in the Tibetan Plateau. The diploid nature and adaptation to diverse environments of the highland give it unique resources for genetic research and crop improvement. Here we produced a 3.89-Gb draft assembly of Tibetan hulless barley with 36,151 predicted protein-coding genes. Comparative analyses revealed the divergence times and synteny between barley and other representative Poaceae genomes. The expansion of the gene family related to stress responses was found in Tibetan hulless barley. Resequencing of 10 barley accessions uncovered high levels of genetic variation in Tibetan wild barley and genetic divergence between Tibetan and non-Tibetan barley genomes. Selective sweep analyses demonstrate adaptive correlations of genes under selection with extensive environmental variables. Our results not only construct a genomic framework for crop improvement but also provide evolutionary insights of highland adaptation of Tibetan hulless barley.

[Effect of electro-acupuncture on hepatic Toll-like receptor 4 and nuclear factor κB expressions in rats with non-alcoholic fatty liver disease].

OBJECTIVE: To investigate the effect of electro-acupuncture on Toll-like receptor 4 (TLR4) nuclear factor κB (NF-κB) expressions in the liver of rats with non-alcoholic fatty liver disease (NAFLD). METHODS: SD rat models of NAFLD induced by feeding high-fat diet for 5 weeks were subjected to early or late intervention with electro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao for 20 min once daily for 21 consecutive days. The changes in serum ALT, AST, TC, and TG levels were determined in the rats and hepatic expressions of TLR4 and NF-κB detected with quantitative fluorescence PCR and Western blotting after treatments. RESULTS: Serum ALT, AST, TC, and TG levels and TLR4 and NF-κB levels increased significantly after NAFLD onset (P<0.05), and were markedly reduced by interventions with electro-acupuncture (P<0.05). CONCLUSION: Electro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao can modify blood lipid and hepatic function to produce positive effect on NAFLD possibly by modifying lipid metabolism, lowering expressions of TLR4 and NF-κB in the liver, and reducing the impact of inflammation on NAFLD.

[Effect of different intensities of electroacupuncture stimulation on expression of SOCS-3 and PPAR-gamma mRNA in adipose tissues of obesity rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of suppressor of cytokine signaling 3 (SOCS-3) and peroxisome proliferator-activated receptor (PPAR)-gamma genes in the epididymis adipose tissue of obesity rats so as to explore its mechanism underlying weight reduction. METHODS: SD rats were randomized into normal control, obesity model, EA-5 mA and EA-1 mA groups (10 rats/group). EA stimulation (6 Hz/20 Hz, 5 mA or 1 mA) was delivered to the bilateral "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) for 15 min, once everyday for two weeks. The expression of SOCS-3 mRNA and PPAR-gamma mRNA in the epididymis adipose tissue was detected by reverse transcriptase polymerase chain reaction. RESULTS: Compared with the normal group, the body weight and expression of SOCS-3 and PPAR-gamma mRNA in the model group were up-regulated obviously (P < 0.01). Compared with the model group, the body weight and expression of SOCS-3 and PPAR-gamma mRNA were significantly decreased in the EA group, with the effects of EA-5 mA being better than those of EA-1 mA. CONCLUSION: EA stimulation of ST 36-SP 6 can reduce body weight in rats with simple obesity, and the effect of higher intensity of EA is apparently better. Accordingly, EA intervention induces down-regulation of SOCS-3 and PPAR-gamma mRNA expression in the epididymis adipose tissue.

[Effect of different intensities of electroacupuncture on expression of monocyte chemoattractant protein-1 and TNF-alpha in adipose tissue in obesity rats].

OBJECTIVE: To observe the effect of different intensities of electroacupuncture (EA) on adipose tissue inflammatory cytokines in rats with simple obesity so as to investigate its mechanism underlying body weight reduction. METHODS: Forty SD male rats were randomly divided into normal, model, strong EA and weak EA groups (n = 10/group). Obesity model was established by feeding the rats with high fat diet. EA (20 Hz, strong EA: 5 V, weak EA: 2. 5 V) was applied to "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) for 15 min, once everyday and for 14 days. Body weight and Lee's index (= body weight(1/3) x 10(3) / body length) were detected. The fasting blood glucose was detected by hexokinase method, serum triglyceride (TG) was detected by glycerol-phosphoric acid oxidase peroxydase (GPO-POD)method, total cholesterol (TC) was detected by cholesterol oxidase method, high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-O) were measured by using one-step method, respectively. The expression of monocyte chemoattractant protein-1 (MOP-1) mRNA and tumor necrosis factor (TNF)-alpha mRNA in the epididymis adipose tissue was detected by reverse transcription-polymerase chain reaction (RT-POR). RESULTS: In comparison with the normal group, the body weight, Lee's index, blood lipid (TG, TC, LDL-C), fasting blood glucose levels, and expression of MOP-1 mRNA and TNF-alpha mRNA were significantly higher in the model group (P < 0.01), and HDL-C was significantly lower (P < 0.01). After EA,compared with the model group, the body weight, Lee's index, TG, TC, LDL-C, fasting blood glucose levels, and expression of MCP-1 mRNA and TNF-a mRNA in both strong and weak EA groups were significantly decreased (P < 0.01, P < 0.05), and HDL-C was significantly increased (P < 0.01, P < 0.05). The effects of strong EA group were obviously superior to those of weak EA group (P < 0.05, P < 0.01). No statistical significance was observed between the two EA groups in fasting blood glucose levels (P > 0.05). CONCLUSION: EA of "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) has a beneficial weight-reduction effect on rats with simple obesity, and moreover, the effect of strong EA stimulation is evidently superior to weak EA stimulation.

[Comparison of volatile oils of cultivated Houttaynia cordata populations with wild].

OBJECTIVE: To provide scientific proofs for the exploitation, utilization, and normalized cultivation of Houttuynia cordata. METHOD: The underground parts of 17 wild H. cordata populations from different valleys and altitudes of mountain. Emei were transplanted to the same growth conditions. After one year's cultivation, volatile oil was obtained by steam distillation from the aerial part of the materials. The chemical constituents were separated and identified by GC-MS, and the relative content of each constituent was determined by area normalization. RESULT: Totally, 31 chemical components were identified, 19 components could be detected in all materials. The t-test results indicated that the contents of alpha-pinene and D-limonene were extremely significantly higher than that in the wild populations, and the contents of camphene and 2-undecanone were also significantly higher than that in the wild populations. And the reverse was found in the content of trans-beta-ocimene. All these 31 components could be divided into 9 chemical compositions, and 7 chemical compositions could be detected in the wild and cultivated. RSD values of 5 chemical compositions in wild populations were higher than that in the cultivated, except for the contents of diterpenyl alcohols and diterpenyl aldehydes. These five chemical compositions accounted for 84.05% and 90.12% of the whole volatile oils in the wild and cultivated conditions, respectively. CONCLUSION: The components and contents of the volatile oils between wild and cultivated were different. The volatile oils polymorphism decrease distinctively as all the wild populations of H. cordata were transplanted to the uniform environmental conditions.