Glucosinolate Biosynthesis Research Articles

Characterisation of the Introgression of Brassica villosa Genome Into Broccoli to Enhance Methionine-Derived Glucosinolates and Associated Health Benefits.

Broccoli cultivars that have enhanced accumulation of methionine-derived glucosinolates have been developed through the introgression of a novel allele of the MYB28 transcription factor from the wild species Brassica villosa. Through a novel k-mer approach, we characterised the extent of the introgression of unique B. villosa genome sequences into high glucosinolate broccoli genotypes. RNAseq analyses indicated that the introgression of the B. villosa MYB28 C2 allele resulted in the enhanced expression of the MYB28 transcription factor, and modified expression of genes associated with sulphate absorption and reduction, and methionine and glucosinolate biosynthesis when compared to standard broccoli. A adenine-thymine (AT) short tandem repeat (STR) was identified within the 5′ untranslated region (UTR) B. villosa MYB28 allele that was absent from two divergent cultivated forms of Brassica oleracea, which may underpin the enhanced expression of B. villosa MYB28.

Synergistic Influence of Sulphur and Boron Fertilization on Enhancing the Productivity of Rapeseed (Brassica napus L.) and Nutrient Status in Subtropical Acidic Soil of Assam, India

ABSTRACT Rapeseed has an immense requirement for S and B, largely for the biosynthesis of glucosinolates and S amino acids. The present investigation was conducted for three consecutive rabi seasons of the year 2014–17 in rapeseed (cv.TS-38) to assess the impact of S and B fertilization on crop yield, soil chemical properties and nutrient status. Results of three years of investigations indicated that among the various soil chemical properties, the combined appliance of S and B level of S30B1.5 (30 kg S ha−1 + 1.5 kg B ha−1) significantly augmented the SOC, while soil pH, Fe2O3 and Al2O3 content in soil were found to be non-affected significantly. Interaction effect between S and B significantly affected available primary nutrients as fertilization with S30B1.5 registered maximum available N (298.7 kg ha−1), P (31.0 kg ha−1) and K (126.2 kg ha−1), respectively. Application of the same levels of S30B1.5 enhanced the status of exchangeable Ca and Mg, available S and B up to 0.90 cmol(p+) kg−1, 0.50 cmol(p+) kg−1, 13.67 mg kg−1 and 0.65 mg kg−1, respectively. The NUE in soil were found to improve on scaling both the levels of S and B. Yield attributing characters were not significantly affected by graded levels of S and B, however, there was a significant improvement in seed yield, stover yield and biological yield of rapeseed. Yield gap analysis exhibited that the combined application of S30B1.0 resulted in the lowest yield gap of 275 kg ha−1 while the control plot recorded the highest yield gap of 647 kg ha−1. Correlation study indicated the existence of a highly significant correlation among most of the soil parameters under investigation. Path analysis revealed that available P was the most dominant soil nutrient implicated in improving the BYR. The inclusive results indicated that higher levels of both S and B could improve the soil fertility status and yield of rapeseed.

Transcriptomic Analysis Reveals Potential Candidate Pathways and Genes Involved in Toxin Biosynthesis in True Toads.

Synthesized chemical defenses have broadly evolved across countless taxa and are important in shaping evolutionary and ecological interactions within ecosystems. However, the underlying genomic mechanisms by which these organisms synthesize and utilize their toxins are relatively unknown. Herein, we use comparative transcriptomics to uncover potential toxin synthesizing genes and pathways, as well as interspecific patterns of toxin synthesizing genes across 10 species of North American true toads (Bufonidae). Upon assembly and annotation of the 10 transcriptomes, we explored patterns of relative gene expression and possible protein-protein interactions across the species to determine what genes and/or pathways may be responsible for toxin synthesis. We also tested our transcriptome dataset for signatures of positive selection to reveal how selection may be acting upon potential toxin producing genes. We assembled high-quality transcriptomes of the bufonid parotoid gland, a tissue not often investigated in other bufonid-related RNAseq studies. We found several genes involved in metabolic and biosynthetic pathways (e.g., steroid biosynthesis, terpenoid backbone biosynthesis, isoquinoline biosynthesis, and glucosinolate biosynthesis) that were functionally enriched and/or relatively expressed across the 10 focal species that may be involved in the synthesis of alkaloid and steroid toxins, as well as other small metabolic compounds that cause distastefulness in bufonids. We hope that our study lays a foundation for future studies to explore the genomic underpinnings and specific pathways of toxin synthesis in toads, as well as at the macroevolutionary scale across numerous taxa that produce their own defensive toxins.

The MIK2/SCOOP Signaling System Contributes to Arabidopsis Resistance Against Herbivory by Modulating Jasmonate and Indole Glucosinolate Biosynthesis.

Initiation of plant immune signaling requires recognition of conserved molecular patterns from microbes and herbivores by plasma membrane-localized pattern recognition receptors. Additionally, plants produce and secrete numerous small peptide hormones, termed phytocytokines, which act as secondary danger signals to modulate immunity. In Arabidopsis, the Brassicae-specific SERINE RICH ENDOGENOUS PEPTIDE (SCOOP) family consists of 14 members that are perceived by the leucine-rich repeat receptor kinase MALE DISCOVERER 1-INTERACTING RECEPTOR LIKE KINASE 2 (MIK2). Recognition of SCOOP peptides elicits generic early signaling responses but knowledge on how and if SCOOPs modulate specific downstream immune defenses is limited. We report here that depletion of MIK2 or the single PROSCOOP12 precursor results in decreased Arabidopsis resistance against the generalist herbivore Spodoptera littoralis but not the specialist Pieris brassicae. Increased performance of S. littoralis on mik2-1 and proscoop12 is accompanied by a diminished accumulation of jasmonic acid, jasmonate-isoleucine and indolic glucosinolates. Additionally, we show transcriptional activation of the PROSCOOP gene family in response to insect herbivory. Our data therefore indicate that perception of endogenous SCOOP peptides by MIK2 modulates the jasmonate pathway and thereby contributes to enhanced defense against a generalist herbivore.

Combined metabolome and transcriptome analysis reveal the mechanism of selenate influence on the growth and quality of cabbage (Brassica oleracea var. capitata L.)

Selenium is an essential trace element for human and animal health, and an appropriate amount of Se can promote the growth and development of plants. Cabbage is a popular cruciferous vegetable with a good ability to accumulate Se, and Se-enriched cabbage can be used as an important Se source for humans. However, the effects of Se-enriched cultivation and the Se accumulation mechanism in cabbage are still unclear. In this study, the effects of different concentrations (0, 0.1, 0.2, 0.4, 0.8, and 1.6 mmol/L) of selenate on cabbage growth and quality were explored. A low concentration of selenate (0.1 mmol/L) promoted growth and nutritional quality. The contents of total Se, S, selenocystine, and selenomethionine significantly increased following selenate application. Important secondary metabolites, namely glucosinolates, phenolic acids, and flavonoids, participate in the response to selenate in cabbage. Comparative transcriptome and metabolomics analysis revealed that SULTR2.2, SULTR3.1, APS, APK2, HMT, MMT, and NTR2 played important roles in Se absorption and conversion. Additionally, the SUR1, UGT74B1, and ST5b genes and cytochrome P450 family genes CYP83A1, CYP79A2, and CYP79F1 may be the crucial genes in the glucosinolates biosynthesis and regulation pathway. The PAL, 4CL, CAD, CHS3, FLS, and CYP73A5 genes were involved in flavonoid and phenolic acid accumulation under selenate treatment. These results reveal the internal relationships in the regulatory network of Se metabolism and secondary metabolite biosynthesis in cabbage and help further the understanding of the physiological and molecular mechanism of how Se biofortification affects cabbage quality, thereby providing genetic resources and a technical basis for the breeding and cultivation of Se-enriched cabbage with excellent nutritional quality.

A Comparative Transcriptome and Metabolome Combined Analysis Reveals the Key Genes and Their Regulatory Model Responsible for Glucoraphasatin Accumulation in Radish Fleshy Taproots.

Radish (Raphanus sativus L.) is rich in specific glucosinolates (GSLs), which benefit human health and special flavor formation. Although the basic GSLs metabolic pathway in Brassicaceae plants is clear, the regulating mechanism for specific glucosinolates content in radish fleshy taproots is not well understood. In this study, we discovered that there was a significant difference in the GSLs profiles and the content of various GSLs components. Glucoraphasatin (GRH) is the most predominant GSL in radish taproots of different genotypes as assessed by HPLC analysis. Further, we compared the taproot transcriptomes of three radish genotypes with high and low GSLs content by employing RNA-seq. Totally, we identified forty-one differentially expressed genes related to GSLs metabolism. Among them, thirteen genes (RsBCAT4, RsIPMDH1, RsMAM1a, RsMAM1b, RsCYP79F1, RsGSTF9, RsGGP1, RsSUR1, RsUGT74C1, RsST5b, RsAPK1, RsGSL-OH, and RsMYB28) were significantly higher co-expressed in the high content genotypes than in low content genotype. Notably, correlation analysis indicated that the expression level of RsMYB28, as an R2R3 transcription factor directly regulating aliphatic glucosinolate biosynthesis, was positively correlated with the GRH content. Co-expression network showed that RsMYB28 probably positively regulated the expression of the above genes, particularly RsSUR1, and consequently the synthesis of GRH. Moreover, the molecular mechanism of the accumulation of this 4-carbon (4C) GSL in radish taproots was explored. This study provides new perspectives on the GSLs accumulation mechanism and genetic improvements in radish taproots.

General regulatory factor 3 regulates the expression of alternative oxidase 1a and the biosynthesis of glucosinolates in cytoplasmic male sterile Brassica juncea

Mitochondrial retrograde signaling (MRS) plays an essential role in sensing and responding to internal and external stimuli to optimize growth to adapt to the prevailing environmental conditions. Previously studies showed alterations on MRS in cytoplasmic male sterile (CMS) plant. However, the regulators involved in MRS in CMS plants remain largely unknown. In this study, we used alternative oxidase 1a (AOX1a) as an indicator of MRS and found that the expression of AOX1a was significantly downregulated in a CMS line comparing to its revertant line, thus indicating an alteration in MRS in the CMS line. By performing a BLAST search of known regulatory components involved in MRS in yeast, we identified general regulatory factor 3 (GRF3), an orthologue of Bmh1/2 in yeast, and demonstrated an association between this gene and MRS in plants, as evidenced by change in AOX1a expression. GRF3 protein was found to be located in the nucleus and the plasma membrane. Further studies showed that GRF3 interacted with MYB29, and regulated the biosynthesis of glucosinolates in Brassica juncea. These findings revealed that GRF3, a negative regulator of AOX1a, is involved in MRS, and also plays a vital role in the accumulation of glucosinolates in CMS crops.

Effect of cultivars on hairy root induction and glucosinolate biosynthesis in a hairy root culture of Kimchi cabbage (Brassica rapa L. ssp. Pekinensis)

Effect of cultivars on hairy root induction and glucosinolate biosynthesis in a hairy root culture of Kimchi cabbage (Brassica rapa L. ssp. Pekinensis)

Melatonin in Brassicaceae: Role in Postharvest and Interesting Phytochemicals.

Brassicaceae plants are of great interest for human consumption due to their wide variety and nutritional qualities. Of the more than 4000 species that make up this family, about a hundred varieties of 6–8 genera are extensively cultivated. One of the most interesting aspects is its high content of glucosinolates, which are plant secondary metabolites with widely demonstrated anti-oncogenic properties that make them healthy. The most relevant Brassicaceae studies related to food and melatonin are examined in this paper. The role of melatonin as a beneficial agent in seedling grown mainly in cabbage and rapeseed and in the postharvest preservation of broccoli is especially analyzed. The beneficial effect of melatonin treatments on the organoleptic properties of these commonly consumed vegetables can be of great interest in the agri-food industry. Melatonin application extends the shelf life of fresh-cut broccoli while maintaining optimal visual and nutritional parameters. In addition, an integrated model indicating the role of melatonin on the organoleptic properties, the biosynthesis of glucosinolates and the regulatory action of these health-relevant compounds with anti-oncogenic activity is presented.

The Investigation of Phenylalanine, Glucosinolate, Benzylisothiocyanate (BITC) and Cyanogenic Glucoside of Papaya Fruits (Carica papaya L. cv. ‘Tainung No. 2’) under Different Development Stages between Seasons and Their Correlation with Bitter Taste

Papaya fruit is one of economic crops in Taiwan, mostly eaten as table fruits. In some Asian countries, unripe papaya fruit is eaten as salad and this led to trends in Taiwan as well. However, unripe papaya fruit may taste bitter during cool seasons. Glucosinolate and cyanogenic glucoside are among the substances that cause bitter taste in many plants, which can also be found in papaya. However, there is still no report about the relationship between seasons and bitter taste in papaya fruits. Thus, the purpose of this study is to investigate the glucosinolate biosynthesis and its correlation between bitterness intensity during cool and warm seasons. The bitterness intensity was highest at the young fruit stage and decreased as it developed. In addition, the bitterness intensity in cool season fruits is higher than in warm season fruits. Cyanogenic glucoside and BITC content showed negative correlation with bitterness intensity (r = −0.54 ***; −0.46 ***). Phenylalanine showed positive correlation with bitterness intensity (r = 0.35 ***), but its content did not reach the bitterness threshold concentration, which suggested that phenylalanine only acts as cyanogenic glucoside and glucosinolate precusors. Glucosinolate content showed positive correlation with bitterness intensity at different developmental stages (r = 0.805 ***). However, the correlation value in different lines/cultivars decreased (0.44 ***), suggesting that glucosinolate was not the only substance that caused bitter taste in immature papaya fruits.

Whole-transcriptome RNA sequencing highlights the molecular mechanisms associated with the maintenance of postharvest quality in broccoli by red LED irradiation

Irradiation of green vegetables with different wavelengths of light represents a safe and effective technology that can be used to maintain postharvest quality. In this regard, our previous research demonstrated that red LED irradiation (50 μmol m−2 s−1) helped to delay the senescence of broccoli stored at room temperature. In the present study, a total of 20,246 differentially expressed mRNAs (DEGs), 3450 differentially expressed lncRNAs (DElncRNAs), and 135 differentially expressed miRNAs (DEmiRNAs) were identified in the response of broccoli to LED irradiation during storage, relative to non-irradiated broccoli. Results of the analysis of the DEG products indicated that the effect of red LED light on delaying senescence in broccoli stored at 20 °C involves an enhancement of genes involved in glucosinolate biosynthesis and the synthesis of chlorophyl a from glutamate. Red LED irradiation also delays increases in the expression of photosynthesis- and oxidative phosphorylation-related genes during the senescence of broccoli. This study provides insight into the molecular mechanisms associated with the response of broccoli to red LED irradiation, and confirms the potential of LED preservation technology.

Identification of Potential Genes Encoding Protein Transporters in Arabidopsis thaliana Glucosinolate (GSL) Metabolism.

Several species in Brassicaceae produce glucosinolates (GSLs) to protect themselves against pests. As demonstrated in A. thaliana, the reallocation of defence compounds, of which GSLs are a major part, is highly dependent on transport processes and serves to protect high-value tissues such as reproductive tissues. This study aimed to identify potential GSL-transporter proteins (TPs) using a network-biology approach. The known A. thaliana GSL genes were retrieved from the literature and pathway databases and searched against several co-expression databases to generate a gene network consisting of 1267 nodes and 14,308 edges. In addition, 1151 co-expressed genes were annotated, integrated, and visualised using relevant bioinformatic tools. Based on three criteria, 21 potential GSL genes encoding TPs were selected. The AST68 and ABCG40 potential GSL TPs were chosen for further investigation because their subcellular localisation is similar to that of known GSL TPs (SULTR1;1 and SULTR1;2) and ABCG36, respectively. However, AST68 was selected for a molecular-docking analysis using AutoDOCK Vina and AutoDOCK 4.2 with the generated 3D model, showing that both domains were well superimposed on the homologs. Both molecular-docking tools calculated good binding-energy values between the sulphate ion and Ser419 and Val172, with the formation of hydrogen bonds and van der Waals interactions, respectively, suggesting that AST68 was one of the sulphate transporters involved in GSL biosynthesis. This finding illustrates the ability to use computational analysis on gene co-expression data to screen and characterise plant TPs on a large scale to comprehensively elucidate GSL metabolism in A. thaliana. Most importantly, newly identified potential GSL transporters can serve as molecular tools in improving the nutritional value of crops.

QM/MM study on enzymatic mechanism in sinigrin biosynthesis

As the major and abundant type of glucosinolates (GL) in plants, sinigrin has potential functions in promoting health and insect defense. The final step in the biosynthesis of sinigrin core structure is highly representative in GL compounds, which corresponds to the process from 3-methylthiopropyl ds-GL to 3-methylthiopropyl GL catalyzed by sulfotransferase (SOT). However, due to the lack of the crystallographic structure of SOT complexed with the 3-methylthiopropyl GL, little is known about this sulfonation process. Fortunately, the crystal structure of SOT 18 from Arabidopsis thaliana (AtSOT18) containing the substance (sinigrin) similar to 3-methylthiopropyl GL has been determined. To understand the enzymatic mechanism, we employed molecular dynamics (MD) simulation and quantum mechanics combined with molecular mechanics (QM/MM) methods to study the conversion from ds-sinigrin to sinigrin catalyzed by AtSOT18. The calculated results demonstrate that the reaction occurs through a concerted dissociative mechanism. Moreover, Lys93, Thr96, Thr97, Tyr130, His155, and two enzyme peptide chains (Pro92-Lys93 and Gln95-Thr96-Thr97) play a role in positioning the substrates and promoting the catalytic reaction by stabilizing the transition state geometry. Particularly, His155 acts as a catalytic base while Lys93 acts as a catalytic acid in the reaction process. The presently proposed concerted dissociative mechanism explains the role of AtSOT18 in sinigrin biosynthesis, and could be instructive for the study of GL biosynthesis catalyzed by other SOTs.

Remodeling and protecting the membrane system to resist phosphorus deficiency in wild soybean (Glycine soja) seedling leaves.

The poor-soil-tolerant wild soybean resist phosphorus deficiency by remodeling membrane lipids to reuse phosphorus. The plants synthesize phenolic acids and flavonoids to remove reactive oxygen species and protect membrane stability. Poor soil largely limits plant yields, and the development and utilization of high-quality wild plant resources is an effective approach to resolving this problem. Two ecotypes of wild soybean were used as experimental materials in this experiment. We integrated metabolomics and transcriptomics to determine whether wild soybean (Glycine soja) could resist phosphorus deficiency by remodeling and protecting its membrane system. Under phosphorus-deficient conditions, the plant height and aboveground fresh and dry weight of poor-soil-tolerant wild soybean seedlings were less inhibited than those in common wild soybean. In poor-soil-tolerant wild soybean seedling leaves, the glycerol-3-phosphate content decreased significantly, while caffeic acid, ferulic acid, shikimic acid, phenylalanine, tyrosine, and tryptophan increased significantly. β-Glucosidase and chalcone synthase genes and those that encode SQD2, a crucial enzyme in thiolipid biosynthesis, were specifically up-regulated, whereas the glucosyltransferase UGT74B1 gene was down-regulated. The poor-soil-tolerant wild soybean enhanced glycerolipid metabolism to decompose phospholipids and release phosphorus for reuse to improve resistance to phosphorus deficiency. The plants synthesized thiolipids to replace phospholipids and maintain membrane structure integrity and inhibited glucosinolate biosynthesis to promote phenylpropanoid biosynthesis, leading to the production of phenolic acids and flavonoids that removed reactive oxygen species and protected membrane system stability. The experiments evaluated and provided insight into the innovative utilization of wild soybean germplasm resources.

A novel putative 2-oxoglutarate-dependent dioxygenase gene (BoaAOP-like) regulates aliphatic glucosinolate biosynthesis in Chinese kale

Glucosinolates are important secondary metabolites in Brassica vegetables, and the side-chain modifications of aliphatic glucosinolates are regulated by 2-oxoglutarate-dependent dioxygenases (AOPs). However, how the side-chain modifications of aliphatic glucosinolates are regulated in Chinese kale remains unclear. Here, we show that the novel gene BoaAOP-like affects the biosynthesis of aliphatic glucosinolates in Chinese kale. BoaAOP-like was localized in the cytoplasm of Chinese kale. The expression levels of BoaAOP-like increased with plant growth; among different organs, BoaAOP-like was the highest in the roots. BoaAOP-like was introduced into Chinese kale using Agrobacterium-mediated transient overexpression system, of which the expression levels were 2.20- to 49.56-fold higher in transiently overexpressed plants than in wild-type plants; the content of total aliphatic glucosinolates in transiently overexpressed plants was up to 70.26-fold higher than in wild-type plants. The content of each aliphatic glucosinolate was significantly accumulated in BoaAOP-like transiently overexpressed plants, with the gluconapin content increasing the greatest when compared to the wild-type plants. The expression levels of aliphatic glucosinolate-related genes, especially MYB28 and AOP2.1 were significantly up-regulated in transiently overexpressed plants. Overall, our results indicated that BoaAOP-like plays a critical role in regulating the biosynthesis of aliphatic glucosinolate in Chinese kale.

Genome-Wide Association Reveals Trait Loci for Seed Glucosinolate Accumulation in Indian Mustard (Brassica juncea L.).

Glucosinolates (GSLs) are sulphur- and nitrogen-containing secondary metabolites implicated in the fitness of Brassicaceae and appreciated for their pungency and health-conferring properties. In Indian mustard (Brassica juncea L.), GSL content and composition are seed-quality-determining traits affecting its economic value. Depending on the end use, i.e., condiment or oil, different GSL levels constitute breeding targets. The genetic control of GSL accumulation in Indian mustard, however, is poorly understood, and current knowledge of GSL biosynthesis and regulation is largely based on Arabidopsis thaliana. A genome-wide association study was carried out to dissect the genetic architecture of total GSL content and the content of two major GSLs, sinigrin and gluconapin, in a diverse panel of 158 Indian mustard lines, which broadly grouped into a South Asia cluster and outside-South-Asia cluster. Using 14,125 single-nucleotide polymorphisms (SNPs) as genotyping input, seven distinct significant associations were discovered for total GSL content, eight associations for sinigrin content and 19 for gluconapin. Close homologues of known GSL structural and regulatory genes were identified as candidate genes in proximity to peak SNPs. Our results provide a comprehensive map of the genetic control of GLS biosynthesis in Indian mustard, including priority targets for further investigation and molecular marker development.

Arabidopsis Glutathione-S-Transferases GSTF11 and GSTU20 Function in Aliphatic Glucosinolate Biosynthesis.

Glutathione (GSH) conjugation with intermediates is required for the biosynthesis of glucosinolate (GSL) by serving as a sulfur supply. Glutathione-S-transferases (GSTs) primarily work on GSH conjugation, suggesting their involvement in GSL metabolism. Although several GSTs, including GSTF11 and GSTU20, have been recently postulated to act in GSL biosynthesis, molecular evidence is lacking. Here, we demonstrated that GSTF11 and GSTU20 play non-redundant, although partially overlapping, roles in aliphatic GSL biosynthesis. In addition, GSTU20 plays a more important role than GSTF11, which is manifested by the greater loss of aliphatic GSLs associated with GSTU20 mutant and a greater number of differentially expressed genes in GSTU20 mutant compared to GSTF11 mutant. Moreover, a double mutation leads to a greater aggregate loss of aliphatic GSLs, suggesting that GSTU20 and GSTF11 may function in GSL biosynthesis in a dosage-dependent manner. Together, our results provide direct evidence that GSTU20 and GSTF11 are critically involved in aliphatic GSL biosynthesis, filling the knowledge gap that has been speculated in recent decades.

Dissecting the Role of SAL1 in Metabolizing the Stress Signaling Molecule 3'-Phosphoadenosine 5'-Phosphate in Different Cell Compartments.

Plants possess the most highly compartmentalized eukaryotic cells. To coordinate their intracellular functions, plastids and the mitochondria are dependent on the flow of information to and from the nuclei, known as retrograde and anterograde signals. One mobile retrograde signaling molecule is the monophosphate 3′-phosphoadenosine 5′-phosphate (PAP), which is mainly produced from 3′-phosphoadenosine 5′-phosphosulfate (PAPS) in the cytosol and regulates the expression of a set of nuclear genes that modulate plant growth in response to biotic and abiotic stresses. The adenosine bisphosphate phosphatase enzyme SAL1 dephosphorylates PAP to AMP in plastids and the mitochondria, but can also rescue sal1 Arabidopsis phenotypes (PAP accumulation, leaf morphology, growth, etc.) when expressed in the cytosol and the nucleus. To understand better the roles of the SAL1 protein in chloroplasts, the mitochondria, nuclei, and the cytosol, we have attempted to complement the sal1 mutant by specifically cargoing the transgenic SAL1 protein to these four cell compartments. Overexpression of SAL1 protein targeted to the nucleus or the mitochondria alone, or co-targeted to chloroplasts and the mitochondria, complemented most aspects of the sal1 phenotypes. Notably, targeting SAL1 to chloroplasts or the cytosol did not effectively rescue the sal1 phenotypes as these transgenic lines accumulated very low levels of SAL1 protein despite overexpressing SAL1 mRNA, suggesting a possibly lower stability of the SAL1 protein in these compartments. The diverse transgenic SAL1 lines exhibited a range of PAP levels. The latter needs to reach certain thresholds in the cell for its impacts on different processes such as leaf growth, regulation of rosette morphology, sulfate homeostasis, and glucosinolate biosynthesis. Collectively, these findings provide an initial platform for further dissection of the role of the SAL1–PAP pathway in different cellular processes under stress conditions.

Vernalization Regulates Flowering Genes and Modulates Glucosinolates Biosynthesis in Chinese Cabbage

Vernalization Regulates Flowering Genes and Modulates Glucosinolates Biosynthesis in Chinese Cabbage

Analysis of Glucosinolate Content and Metabolism Related Genes in Different Parts of Chinese Flowering Cabbage.

Glucosinolates (GSLs) are important secondary metabolites that play important defensive roles in cruciferous plants. Chinese flowering cabbage, one of the most common vegetable crops, is rich in GSLs and thus has the potential to reduce the risk of cancer in humans. Many genes that are involved in GSL biosynthesis and metabolism have been identified in the model plant Arabidopsis thaliana; however, few studies investigated the genes related to GSL biosynthesis and metabolism in Chinese flowering cabbage. In the present study, the GSL composition and content in three different organs of Chinese flowering cabbage (leaf, stalk, and flower bud) were determined. Our results showed that the total GSL content in flower buds was significantly higher than in stalks and leaves, and aliphatic GSLs were the most abundant GSL type. To understand the molecular mechanisms underlying the variations of GSL content, we analyzed the expression of genes encoding enzymes involved in GSL biosynthesis and transport in different tissues of Chinese flowering cabbage using RNA sequencing; the expression levels of most genes were found to be consistent with the pattern of total GSL content. Correlation and consistency analysis of differentially expressed genes from different organs with the GSL content revealed that seven genes (Bra029966, Bra012640, Bra016787, Bra011761, Bra006830, Bra011759, and Bra029248) were positively correlated with GSL content. These findings provide a molecular basis for further elucidating GSL biosynthesis and transport in Chinese flowering cabbage.