Erinacine A, a cyathane-type diterpenoid mainly from Hericium erinaceus mycelia, exhibits prominent neurotrophic and neuroprotective activities, making it a promising candidate for managing neurodegenerative diseases. However, its low abundance and unclear genetic regulatory mechanisms hinder its application as a nutraceutical. This study aimed to decipher its regulatory mechanisms and enhance production. Four exogenous inducers were screened, with salicylic acid (SA) and ergosterol (ERG) significantly increasing erinacine A content by 62.21% and 146.70% at 20 days, respectively. Transcriptome and WGCNA of inducer-treated sample identified darkorange and magenta modules associated with erinacine A biosynthesis, with the eri gene cluster enriched in the darkorange module and eriG and eriF as hub genes. Forward genetic analysis via QTL mapping of the HeD127 dikaryon population revealed significant phenotypic variation in erinacine A content (0.341-13.085 mg/g) and identified two loci (erA-1 and erA-2) explaining 18.63% of phenotypic variation. Integrating these forward and reverse genetic analyses revealed that salicylic acid and ergosterol synergistically regulate core carbon metabolic pathways to augment acetyl-CoA supply for the mevalonate pathway, suppressed competitive metabolism, enhanced diterpene skeleton construction and structural modification. These results deepen our understanding of the genetic and molecular basis governing accumulation of erinacine A, and facilitate its application in neuroprotective pharmaceuticals.

Sulforaphane (SFN) is widely recognized for its health-promoting benefits. While several studies have reported that methyl jasmonate (MeJA) enhances SFN accumulation, the role of epigenetic regulation remains unclear. Here, we performed an integrated analysis of transcriptome, whole-genome bisulfite sequencing, and small RNA-seq data from broccoli florets and leaves under MeJA treatment and control conditions. Our results revealed that MeJA-responsive differentially methylated regions (DMRs) predominantly occurring in the CHG context. Integrative analysis of methylation and siRNA data indicated that changes in MeJA-responsive CHH methylation were associated with 24-nucleotide siRNA clusters. Furthermore, inhibiting DNA methylation using 5-azacytidine treatment in broccoli seedlings increased SFN content, validating the role of DNA hypomethylation in this process. By combining transcriptional regulatory network analysis with DMR data, we identified the MeJA-responsive CHG-hypomethylated gene BoCYP83A1. The McrBC-PCR assay confirmed reduced methylation in the BoCYP83A1 promoter region. The function of BoCYP83A1 in MeJA-induced SFN biosynthesis was validated using RNA interference in broccoli. Further evidence demonstrated that the transcription factors BoMYB68 and BoIAA18 promote BoCYP83A1 expression by directly binding to its promoter. Our findings not only provide novel insights into the epigenetic regulation underlying MeJA-induced SFN accumulation in plants but also facilitate molecular breeding for the development of high-SFN crops.

Ergosterol, a precursor of vitamin D₂ and a substrate for steroid drug synthesis, is a key metabolite in macrofungi. In this study, Trametes versicolor PS genome (58.86 Mb) was assembled using HiFi sequencing and 79 genes involved in ergosterol biosynthetic pathways were annotated. Six ERG25 family members (TvERG25-1 ~ 6) were identified, encoding proteins of 327 ~ 372 amino acids with 37,617 ~ 42,809 Da MW. TvERG25-1 and TvERG25-4 displayed highly similar gene architectures, and together with TvERG25-3, shared conserved motif and clustered within subfamily II, whereas TvERG25-2 and TvERG25-6 contained fewer conserved motifs. Light-responsive elements were abundant in six genes. HPLC analysis revealed ergosterol level in T. versicolor mycelia increased under white, blue, and green light stress (P < 0.001), peaking under blue light (2076.0 ± 1.1 μg/g). Ergosterol levels correlated positively with TvERG25-2, TvERG25-4, and TvERG25-6 expression. This study establishes a basis for elucidating TvERG25 function and for developing T. versicolor strains with enhanced ergosterol content.

Purple flowering stalk, with vivid color and high yield, is a special vegetable rich in glucosinolates. This study analyzed and evaluated the variability of agronomic traits and nutritional quality of 127 genotypes of purple flowering stalks. The results showed that the genetic diversity index of qualitative traits varied from 0.16 (leaf vein clarity) to 1.57 (cotyledon color), and quantitative traits varied from 1.48 (rosette number) to 2.06 (cotyledon groove depth). Moreover, the highest coefficient of variation in quantitative traits was found in three yield-related traits: main flowering stalk weight, main flowering stalk length, and lateral flowering stalk number (63.35 %, 45.17 %, and 46.57 %, respectively). Cluster analysis divided accessions into three major clusters. The hierarchical cluster analysis implied that the yield-related traits and nutritional traits contribute most to the distinction. In principal component analysis based on yield and nutritional traits, the eight components explained 76.4 % of the variance in the data. All varieties were evaluated by comprehensive score, and the top ten of them (CS 70, CS 56, CS 105, CS 114, CS 113, CS 129, CS 93, CS134, CS 35, CS 74) with different characters can be used in the improvement of breeding. Correlation analysis suggested that shorter flowering stalks may have more content of total glucosinolates, ascorbic acid, and soluble protein. In conclusion, this work provides basic materials and theories for purple flowering stalk breeding.

Sulforaphane (SF) is a sulfur (S)-containing isothiocyanate found in cruciferous vegetables and is known for its potent anticancer properties. Broccoli sprouts, in particular, are considered safe and healthy dietary choices due to their high SF content and other beneficial biological activities, such as enhanced metabolite ingestion. The application of selenium (Se) is an excellent approach to enhance the abundance of SF. Previous studies have often focused on gene expression and changes in the synthetic substrates of glucoraphanin (RAA) to explain SF variation in response to Se application. However, the regulatory network and other physiological and biochemical reactions involved in the regulation of SF biosynthesis are poorly understood. In this study, Se-treated broccoli sprouts had higher SF and RAA contents; they increased with increasing Se application. Using RNA-seq in combination with KEGG, GO, phenotypic, and WGCNA analyses, it was observed that not only gene expression was induced but also that glutathione serves as an S donor for SF biosynthesis and acts as an oxidative stress reliever as a result of Se treatment. Additionally, a module related to glucosinolate biosynthesis was identified. Yeast one-hybrid system and dual luciferase reporter assay were utilized. These assays demonstrated the hub transcription factors GATA22, ERF12-like, and MYB108 would directly bind to SUR1 promoter and positively regulate its expression. Our study presents the first global overview of the role of GSH metabolism in response to Se for SF biosynthesis, and provides a novel and valuable gene resource for the molecular breeding of high-SF broccoli.

Broccoli is an economically significant vegetable with high nutritional and medicinal value. Seed size/weight is one of the important agronomic traits that determine crop yield, which is regulated by multiple plant hormones. However, limited information is known about the regulation of seed size control in broccoli. Here, we report that the basic helix-loop-helix transcription factor BoMYC2 exerts a negative regulatory influence on seed-related traits in broccoli. The overexpression of BoMYC2 in Arabidopsis and broccoli led to notably decreased seed size and seed weight, likely by repressing cell proliferation of embryonic. Besides, overexpression of BoMYC2 increased the fatty acid accumulation while reducing the protein and soluble sugar levels in stably transgenic broccoli seeds, affecting seed storage compounds composition and seed size. Based on DNA affinity purification sequencing, BoMYC2 targeted the cytokinin oxidase gene BoCKX5 by binding to the G-box motif in its promoter regions. BoMYC2 activated the expression of BoCKX5 to modulate cytokinin metabolism. The expression levels of cytokinin-related genes and jasmonic acid-related genes were further analyzed in transgenic broccoli seeds, implying that BoMYC2 leads to higher jasmonic acid and lower cytokinin hormone content, which might affect the cell proliferation in seeds. Furthermore, cytokinin levels were significantly lower in BoMYC2-overexpressing broccoli than in wild-type plants. Collectively, these findings reveal a BoMYC2-BoCKX5 regulatory module that controls seed size/weight, enriching our understanding of the internal mechanism in seed size regulation and providing promising targets for high-yield broccoli breeding.

Sulforaphane, a naturally specialized metabolite, plays significant roles in human disease prevention and plant defense. Myrosinase (MY) is a key gene responsible for the catalysis of sulforaphane formation, but the molecular mechanisms through which MY regulates sulforaphane biosynthesis in plants remains largely unknown. Here, we discovered that the change of sulforaphane content in broccoli sprouts caused by exogenous selenite treatments is positively related to BoMY expression. BoMY overexpression in the Arabidopsis thaliana tgg1 mutants could dramatically increase myrosinase activity and sulforaphane content in the rosette leaves of 35S::BoMY/tgg1 and rescue its phenotypes. Moreover, an obvious increase of myrosinase activity and sulforaphane content was displayed in transgenic BoMY-overexpressed broccoli lines. In addition, a 2 033 bp promoter fragment of BoMY was isolated. Yeast one-hybrid (Y1H) library screening experiment uncovered that one bHLH transcription factor, BoFAMA, could directly bind to BoMY promoter to activate its expression, which was further evidenced by Y1H assay and dual-luciferase reporter assay. BoFAMA is a selenite-responsive transcription factor that is highly expressed in broccoli leaves; its protein is solely localized to nucleus. Additionally, genetic evidence suggested that the knockdown of FAMA gene in Arabidopsis thaliana could significantly decrease sulforaphane yield by inhibiting the expression of myrosinase genes. Interestingly, exogenous selenite supply could partially restore the low level of sulforaphane content in transgenic Arabidopsis FAMA-silencing plants. Our findings uncover a novel function of FAMA-MY module in the regulation of selenite-mediated sulforaphane synthesis and provide a new insights into the molecular mechanism by which selenite regulates the accumulation of sulforaphane in plants.

Selenocysteine methyltransferase (SMT) is a key enzyme involved in the Se metabolism pathway, and it is responsible for the catalysis of Se-methylselenocysteine (SeMSC) compound formation. Previous studies showed that selenium treatment activated SMT expression and promoted the accumulation of glucosinolates (GSLs) and sulforaphane, but the roles and functional mechanisms of SMT in mediating GSLs and sulforaphane synthesis remain unclear. In this study, we identified the BoSMT gene in broccoli and uncovered its roles in mediating GSLs biosynthesis. Transgenic assays revealed that BoSMT is involved in SeMSC biosynthesis in broccoli. More importantly, the contents of GSLs and sulforaphane were significantly increased in the BoSMT-overexpressing broccoli lines but decreased in the knockdown lines, suggesting that BoSMT played a positive role in regulating GSLs and sulforaphane synthesis. Further evidence indicated that BoSMT-mediated overaccumulation of GSLs and sulforaphane might be due to the increase in the endogenous SeMSC content. Compared with the mock (water) treatment, selenite-induced significantly increases of the SeMSC content in the BoSMT-knockdown plants partially compensated the phenotype of GSLs and sulforaphane loss. Compared with the mock treatment, exogenous SeMSC treatment significantly increased the contents of GSL and sulforaphane and activated GSL synthesis-related gene expression, suggesting that SeMSC acted as a positive regulator for GSL and sulforaphane production. Our findings provided novel insights into selenium-mediated GSLs and sulforaphane accumulation. The genetic manipulation of BoSMT might be a useful strategy for improving the dietary nutritional values of broccoli.

Broccoli (Brassica oleracea var. italica Plenck) is an important vegetable crop, as it is rich in health-beneficial glucosinolates (GSLs). However, the genetic basis of the GSL diversity in Brassicaceae remains unclear. Here we report a chromosome-level genome assembly of broccoli generated using PacBio HiFi reads and Hi-C technology. The final genome assembly is 613.79 Mb in size, with a contig N50 of 14.70 Mb. The GSL profile and content analysis of different B. oleracea varieties, combined with a phylogenetic tree analysis, sequence alignment, and the construction of a 3D model of the methylthioalkylmalate synthase 1 (MAM1) protein, revealed that the gene copy number and amino acid sequence variation both contributed to the diversity of GSL biosynthesis in B. oleracea. The overexpression of BoMAM1 (BolI0108790) in broccoli resulted in high accumulation and a high ratio of C4-GSLs, demonstrating that BoMAM1 is the key enzyme in C4-GSL biosynthesis. These results provide valuable insights for future genetic studies and nutritive component applications of Brassica crops.

为探究以南荻为基质栽培秀珍菇的效果,试验以台秀1号为试验菌株,考察南荻基质含量(16%、32%、48%、64%、80%)对台秀1号菌丝生长速度、产量以及营养品质的影响.试验结果表明,栽培料中添加南荻基质,台秀1号菌丝生长比对照快,且菌丝浓密;配方①、配方②与对照(CK)的产量相当,配方③、配方④、配方⑤与对照产量差异显著.南荻32%,棉籽壳30%,杂木屑18%,麸皮15%,石灰1%,石膏1%,糖1%、玉米粉2%的配方②栽培秀珍菇的效果最好.

Using the whole genome sequence of PM＿ss5 downloaded from GenBank database, genome-wide simple sequence repeats（SSRs） of Pleurotus pulmonarius were mined and characterized. A total of 2348 SSR loci were detected in PM＿ss5 genome, with a relative abundance of 59 SSRs per Mb. Among all SSR loci, the di-nucleotide repeats were the dominant（51.8%）, followed by the tri-nucleotide repeats（27.7%）. The identified SSR loci contained 141 base motifs,among which the dominant base motifs were GA/TC and CT/AG. The length of SSRs ranged from 10 to 156 bp, and 77.2% of the identified SSRs were 10 to 15 bp. The SSR patterns in P. pulmonarius were also compared with those of other four species of Pleurotus. The short nucleotide repeats were dominant SSRs in all five Pleurotus spp., with the di-nucleotide SSRs accounting for higher rate in P. pulmonarius. Then, 53 pairs of polymorphic SSR primers were designed and selected to analyze the genetic diversity of the 18 varieties of P. pulmonarius, which showed P. pulmonarius owned moderate genetic diversity, and the average numbers of effective alleles, Nei’s genetic diversity and Shannon index were 1.35, 0.23 and 0.38, respectively.

Vegetable crops are abundant in a variety of biologically active compounds that have biological effects on human metabolism, immunity, and the prevention of cancer and chronic diseases. Light is one of the environmental elements that has the greatest impact on the growth and development of vegetable crops. It also plays a significant role in controlling the synthesis and metabolism of a variety of bioactive chemicals in plants. The molecular mechanism of light quality regulating the aforementioned active substances was summarized. This article analyzes the effects of LED light quality and ratio on the metabolism of active substances such as phenolic compounds, carotenoid, sulforaphane, and vitamin C in vegetable crops. In addition to serving as a theoretical guide, this review can offer technical assistance for LED light quality control of high-quality vegetables as well as theoretical support for the measurement of bioactive compounds in vegetable crops.

Tobacco mosaic virus（TMV） is a major factor leading to the yield reduction of Solanaceae crops. In order to explore edible fungus-derived polysaccharide materials for the prevention and control of Tobacco mosaic virus, the control effect of six crude polysaccharides against TMV was determined by the half-leaf method, five-point injection test and systematic host infection activity evaluation. Among them, the extraction rate of crude polysaccharide from Agaricus blazei, 24.32%, was the highest, followed by Pleurotus ostreatus 18.56%, and that from Pleurotus eryngii,10.04% was the lowest. The total sugar content of A. blazei was significantly higher than that of other species, reaching 37.71%, followed by that of P. igniarius and G. Lucidum, exceeding 32.00%. In addition, the sulfate content of A. blazei crude polysaccharide was also as high as 2.10%. In this research, the inhibitory effect of the crude polysaccharides was determined by the half leaf method. Among the 6 tested polysaccharide treatments, the polysaccharides that were inoculated after treated for 48 h had a good inhibition on TMV. The preventive effects of P. ostreatus, C. versicolor and A. blazei reached more than 60.00%. Especially, the preventive effect of 24 mg/mL A. blazei crude polysaccharide was up to 92.04%, and the number of dead spots on the leaves was significantly less than those in the clear water control group. In the five-point injection test, the inhibition rate of A. blazei crude polysaccharide was 38.18%. The brightness of the fluorescent spots was weaker than that in the control group, the fluorescent area did not change much, and did not appeared in the new leaves. A. blazei crude polysaccharide had an inhibitory effect in the early stage of virus infection,which could delay the moving speed of TMV and slow down the system invasion process. On NC-89, the control effect of agaricus antler polysaccharide on TMV was as high as 76.81%, which was better than that of 0.5% lentinan water agent and 1.8% octinamine acetate water agent. When the concentration was 48 mg/mL, the plants were damaged, and the excessive concentration of polysaccharide formed precipitation on the surface of the plants, causing the plants to yellow. Field test showed that the treatment effect of A. blazei crude polysaccharides on TMV was 70.31%. The plants with twisted, curled leaves and stopped growing had an obvious improvement. At the same time, it had a certain inhibitory effect on TMV, the onset time of sprayed tobacco was delayed, the tobacco grew well, and the leaves were extended. The results of this study would lay a foundation for further study on the action mode and mechanism of fungal polysaccharides against Tobacco mosaic virus.

Chinese broccoli [Brassica oleracea var. alboglabra (L. H. Bailey) Musil] is an annual herbaceous healthy vegetable plant mainly cultivated in southern China. Previous studies have been primarily focused on investigating the nutritional ingredients of this plant, little is known about its medicinal potentials. Here we report the profiling of sulforaphane, an active ingredient with potent anticancer activities, in 47 Chinese broccoli germplasms collected from China and some southeast-Asian countries. High-performance liquid chromatography technology was used to systemically quantify the sulforaphane content in leaf, stem, and flower bud clusters of all 47 Chinese broccoli germplasms. The results indicated a diverse distribution pattern of sulforaphane among different tissues. Flower bud contained the highest average of sulforaphane, roughly twice as much as in leaf and stem tissues. Interestingly, different germplasms exhibited great differences in terms of sulforaphane abundance. Germplasm BOK15-20 is extremely rich in sulforaphane in all tissues tested, similar to 2,500 times of BOK15-15 in leaf, 66 times of BOK15-42 in the stem, and 77 times of BOK15-40 in flower bud. The tremendous disparity of sulforaphane content among different germplasms suggests considerable variations in germplasms' medicinal potentials. In addition, we systemically characterized a series of morphological and phenotypic traits of all 47 Chinese broccoli germplasms. Cluster analysis through NTSYSpc v2.2 showed that the 47 germplasms were classified into five different groups, respectively, based on sulforaphane content and qualitative phenotypic traits. Our results not only confirmed the medicinal values of Chinese broccoli but also provided crucial information that will eventually benefit future germplasm development and new variety breeding.

为探寻合理的化肥与有机肥配施方式,改善青花菜生长、品质、土壤状况及经济效益,以国王100青花菜品种为材料,采用单因素随机区组设计,以不施用肥料的CK和常规施用化肥的CF为对照,设置T200(化肥减施25％+增施200 kg·667 m-2有机肥)和T400(化肥减施25％+增施400 kg·667 m-2有机肥)2个处理.结果表明,T200、T400处理分别较CF每667 m2增产126.23、61.71 kg;T200处理下可溶性糖含量最高,为7.48 mg·g-1;青花菜对钾肥的吸收利用率在T200处理下达到最高,且显著高于其他处理.综上所述,在T200(化肥减施25％+增施200 kg·667 m-2有机肥处理)可以提高青花菜品质、增强肥料有效性、改善土壤养分环境、增加农民收益,是一种适于生产需要、提升青花菜种植经济效益的合理施肥方式.

Mustard, which belongs to the family Brassicaceae, is an annual or biennial herb and is considered as one of the most important native vegetables in China. Glucosinolates are important secondary metabolites containing sulfur and nitrogen in plants, which form a network with other metabolic pathways that play important roles in plant growth, development, and interaction with the environment. We studied varied phenotypic and glucosinolate contents of 60 mustard resources collected from various areas of China. The results showed both agronomic traits and glucosinolates varied greatly among mustard resources. We detected nine glucosinolates in mustard resources and the contents of total glucosinolates ranged from 1.2023 to 30.7310 μmol/g. Through the correlation analysis, we preliminarily found a significant negative correlation between leaf color and glucosinolate contents but needed further validation. For mustard resource JC 18-56, we analyzed the glucosinolate contents in different organs of different growth stages. The results indicated a significant difference among organs in both glucosinolates concentration and composition. The contents of glucosinolatess in alabastrums at bolting stage were highest, up to 140.1257 μmol/g dry weight (DW). We found that the main glucosinolates in roots were 4-methoxyglucobrassicin, while in other organs the glucosinolates were sinigrin. The contents of glucosinolatess in different organs of mustard were as follows: alabastrums > seeds > flowers > siliques > leaves > flower stems > stems > roots. This study provides important references for the selection and cultivation of high-quality mustard varieties.

Broccoli is becoming increasingly popular among consumers owing to its nutritional value and rich bioactive compounds, such glucosinolates (GSLs) and hydrolysis products, which are secondary metabolites for plant defense, cancer prevention, and higher antioxidant activity for humans. In this study, 40 μmol/L methyl jasmonate (MeJA) was sprayed onto broccoli from budding until harvest. The harvested broccoli florets, stem, and leaves were used to measure the contents of GSLs, sulforaphane, total phenolics, and flavonoids, as well as myrosinase activity, antioxidant activity, and gene expression involved in GSL biosynthesis. The overall results revealed that GSL biosynthesis and sulforaphane accumulation were most likely induced by exogenous MeJA treatment by upregulating the expression of CYP83A1, SUR1, UGT74B1, and SOT18 genes. Exogenous MeJA treatment more remarkably contributed to the increased GSL biosynthesis in broccoli cultivars with low-level GSL content (Yanxiu) than that with high-level GSLs (Xianglv No.3). Moreover, MeJA treatment had a more remarkable increasing effect in broccoli florets than stem and leaves. Interestingly, total flavonoid content substantially increased in broccoli florets after MeJA treatment, but total phenolics did not. Similarly, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, trolox-equivalent antioxidant capacity (ABTS), and ferric-reducing antioxidant power (FRAP) were higher in broccoli floret after MeJA treatment. In conclusion, MeJA mediated bioactive compound metabolism, had positive effects on GSL biosynthesis, sulforaphane, and flavonoids accumulation, and showed positive correlation on inducing higher antioxidant activities in broccoli floret. Hence, preharvest supplementation with 40 μM MeJA could be a good way to improve the nutritional value of broccoli florets.

夏季高温炎热、暴雨、强光照,严重影响越夏辣椒育苗,为探究不同颜色和密度的遮阳网对越夏辣椒幼苗生长的影响,以软皮15号辣椒品种为试材,分别以5种不同规格的遮阳网:绿色3针、黑色3针、绿色6针、黑色6针、蓝色6针覆盖育苗,以不覆盖遮阳网为对照.结果表明,黑色3针遮阳网处理的辣椒幼苗株高、地上部鲜质量及叶片叶绿素、类胡萝卜素含量、初始荧光(Fo)、最大荧光(Fm)、最大光化学效率(Fv/Fm)、光化学猝灭系数(qP)均显著高于对照,提高了越夏辣椒幼苗叶片生长及同化产物向营养器官的运输和积累、光合色素含量、PSⅡ反应中心的开放程度,保护光合系统膜完整、光合电子传递顺畅.结合各项指标,认为黑色3针遮阳网覆盖有利于越夏辣椒育苗,值得在生产上推荐.