Comparative transcriptome analyses of genes involved in sulforaphane metabolism at different treatment in Chinese kale using full-length transcriptome sequencing

- genes involved in sulforaphane metabolism at different treatment in Chinese kale using full-length transcriptome sequencing.
- Here, we investigated the molecular mechanism related to glucosinolates biosynthesis in Chinese kale using combined NGS and SMRT sequencing..
- NaCl (75 mM), methyl jasmonate (MeJA, 40 μ M), selenate (Se, sodium selenite 100 μ M), and brassinolide (BR, 1.5 μ M) treatment induced and 11,041 differentially expressed genes (DEGs) in Chinese kale seedlings comparing with control.
- These genes were associated with pathways of glucosinolates biosynthesis, including phenylalanine, tyrosine and tryptophan biosynthesis, cysteine and methionine metabolism, and glucosinolate biosynthesis.
- We found NaCl decreased sulforaphane and glucosinolates (indolic and aliphatic) contents and downregulated expression of cytochrome P45083b1 (CYP83b1), S-alkyl-thiohydroximatelyase or carbon – sulfur lyase (SUR1) and UDP-glycosyltransferase 74B1 (UGT74b1).
- MeJA increased sulforaphane and glucosinolates contents and upregulated the expression of CYP83b1, SUR1 and UGT74b1.
- Conclusions: We confirmed that NaCl inhibited the biosynthesis of both indolic and aliphatic glucosinolates, while MeJA and BR increased them.
- MeJA and BR treatments, conferred the biosynthesis of glucosinolates, and Se and MeJA contributed to sulforaphane in Chinese kale via regulating the expression of CYP83b1, SUR1 and UGT74b1..
- NaCl reduced sulforaphane and glucosinolate contents in Chinese kale..
- MeJA, BR, and Se increased sulforaphane and/or glucosinolate contents..
- the administration of sulforaphane shows antitumor formation [9].
- The biosynthesis and ac- cumulation of glucosinolates as well as sulforaphane is influenced by temperature, processing methods and abiotic stresses .
- They confirmed that NaCl and SA co-treatment induced sulforaphane and isothiocyanates accumulation was inde- pendent of the biosynthetic genes of cytochrome P45083b1 (CYP83b1), CYP83a1 and CYP79b2, but associ- ated with the increased hydrolytic enzymes of myrosinase (MYO) and epithiospecifier moifier 1 (ESM1) [17].
- performed a comparative transcriptome analysis in two broccoli cultivars under MeJA treatment and iden- tified that the glucosinolates’ core structure biosynthesis genes (including CYP79b2, UDP-glycosyltransferase 74B1, UGT74b1, SUR1) were upregulated by MeJA [23].
- Several comparative transcriptome analyses in Chinese kale (B..
- alboglabra Bailey) [24, 25] and watercress [26] showed that the expression patterns of aforemen- tioned genes related to glucosinolates biosynthesis were changed in different tissues.
- It has been reported that the influence of stress-induced bio- synthesis of sulforaphane is dependent on plant variety and growth conditions [17, 27].
- However, the causality of abiotic stresses-induced dysregulation of sulforaphane and glucosinolates biosynthesis has yet to be clearly explored, and the underlying mechanisms should be dissected..
- Here we investigated the molecular mecha- nisms related to glucosinolates biosynthesis under different abiotic stresses in Chinese kale using combined NGS and SMRT sequencing data.
- The market demand of Chinese kale sprouts is increasing due to the rich in glu- cosinolates.
- Transcriptome with complete and full-length reads would be to dissect molecular mechanisms of gluco- sinolates biosynthesis in Chinese kale..
- Sulforaphane and glucosinolates contents were changed by different treatments.
- We firstly treated with Chinese kale seedlings with series of NaCl, Se, MeJA and BR to select the optimal.
- Treatment with NaCl (0–150 mM) and BR (0–3.0 μM) significantly reduced sulforaphane..
- We confirmed that the contents of sulforaphane (Fig..
- 1c, total, aliphatic and indo- lic) in Chinese kale seedlings were significantly reduced by NaCl treatment.
- Se and MeJA treatments obviously increased the contents of sulforaphane in kale seedlings in comparison with control (p <.
- 3a), and MeJA and BR treatments significantly increased the contents of total and aliphatic glucosinolates (p <.
- 1 The content of sulforaphane and glucosinolates in Chinese kale seedlings in response to different treatments.
- a the content of sulforaphane in Chinese kale seedlings under different treatments with series concentrations.
- b the content of sulforaphane under different treatments with optimal contents.
- c the total content of 18, aliphatic, aromatic and indolic glucosinolates in Chinese kale seedlings (dry weight, g).
- Summary of Illumina Hiseq and PacBio RSII transcriptome sequencing data of Chinese kale.
- In comparative analysis, we observed that the unigene number originated from PacBio RSII transcriptome (185,134 genes) was higher than that of Illumina tran- scriptome (130,553 genes.
- KEGG and KOG classification showed that the top clusters involved uni- genes were energy and carbohydrate metabolism, trans- port, catabolism and signal transduction (Additional file 3 Figure S3B and C).
- followed with Brassica napus (38.0%) and Brassica rapa (3.0.
- while Table 1 Summary of the transcriptome data from PacBio RSII platform and Illumina Hiseq platform.
- PacBio RSII (n = 15).
- Additional file 3: Figure S3D)..
- A total of 6893 (includ- ing 3176 up- and 3717 down-regulated DEGs), 13,287 (including 4377 up- and 8910 down-regulated DEGs), 13,659 (including 4879 up- and 8780 down-regulated DEGs), and 11,041 DEGs (including 4351 up- and 6690 down-regulated DEGs) were identified from Chinese kale seedlings under NaCl, Se, MeJA, and BR treatment, respectively (p-value.
- Analysis of DEGs in pathways related to glucosinolate biosynthesis.
- According to aforementioned results, we confirmed that the influence of NaCl, BR, MeJA, and Se treatments on glucosinolates accumulation might be mediated by.
- altering the expression profiles of sulforaphane and/or glucosinolate biosynthesis related genes.
- CDY01177.1) and HMT1 (homocysteine S-methyltransfer- ase 1.
- CDY19364.1], DNMT2 (CDX99400.1) and metE (5-methyltetrahydropteroyltriglutamate:homocysteineme- thyltransferase 1.
- CDX85436.1) were increased by Se, MeJA and BR treatments.
- the expression of CYP83b1 (CDX80735.1 and XP CYP79b1 (CDX69373.1), SUR1 (XM and UDP- glycosyltransferase 74B1 (UGT74b1.
- ACB59204.1) were up- regulated by MeJA and BR (p <.
- AAP92453.1) and SAHH2L X4 (adenosylhomocysteinase 2 (LOC transcript variant X4.
- The glucosinolate biosynthesis pathway and DEGs induced by Se, NaCl, MeJA and BR are shown in Additional file 6: Figure S4.
- c clustering analysis of the DEGs.
- ERF12), SNF2 family, and WRKY family (including WRKY33, WRKY18, WRKY48, WRKY51, WRKY22–1), and upregulated MYB28 and WRKY47.
- We performed the combined transcriptome analysis of NGS and SMRT sequencing and investigated the molecular mechanism of sulforaphane and glucosin- olate biosynthesis in Chinese kale seedlings in response to abiotic stresses.
- The combination of NGS short reads and SMRT long reads identified and 11,041 DEGs in kale seedlings in response to NaCl (75 mM), MeJA (40 μM), Se (100 μM) and BR (1.5 μM) treatment, re- spectively.
- These pathways were core pathways for the synthesis of glucosinolates (Additional file 6: Figure S4).
- These facts showed that these biosyn- thetic pathways were influenced by NaCl, MeJA, Se and BR treatments in Chinese kale seedlings, and the gluco- sinolates might take important roles in the plant defense or resistance to these abiotic stresses..
- We confirmed that the content of aliphatic glucosinolates was highest in Chin- ese kale seedlings, followed with indolic glucosinolates and then aromatic glucosinolates.
- In addition, we con- firmed that the content of aliphatic glucosinolates was increased by MeJA and BR treatments, while aromatic glucosinolates was increased by Se and BR treatments (Fig.
- Transcriptome analysis showed that the expres- sion of genes in core pathways of aliphatic (including CYP83a1, SUR1, and UGT74b1) and aromatic glucosin- olates (including CYP83b1, SUR1 and UGT74b1), were increased by MeJA and BR treatments (Fig.
- In addition, we found the con- tents of sulforaphane and total glucosinolates were increased by MeJA and BR treatments compared with control.
- We also observed the dec- rement in ST5a_b_c expression by Se, MeJA and BR.
- It has been reported that the synthesis of aliphatic gluco- sinolates could be controlled by ST5b_c, and the aro- matic glucosinolates is controlled by ST5a (Additional file 6: Figure S4).
- ordinal mechanism of glucosinolates biosynthesis in plant [33]..
- Our current study confirmed that the contents of all glucosinolates and sulforaphane in Chinese kale were decreased by NaCl stress (75 mM).
- They also suggested that the increased contents of sulfo- raphane by NaCl stress (80 and 160 mM) were associ- ated with upregulation of MYO and ESM1.
- showed the higher expression of MYO was found in Chinese kale roots with low-glucosinolates [25], which might responsible for the increased sulforaphane [17]..
- Our study demonstrated that the expression of CYP83a1 and CYP83b1 were not significantly influenced by 75 mM NaCl stress.
- In addition, the treatment with 75 mM NaCl decreased the contents of both sulforaphane and glucosinolates.
- Although total and the major aliphatic glucosinolates were increased by MeJA and BR treat- ments, the FPKM of ST5a (crucial for aromactic gluco- sinolates) and ST5b_c (crucial for aliphatic.
- Additional file 6: Figure S4) coding se- quences were decreased by MeJA and BR compared with control (Additional file 7: Figure S5).
- Other studies of Chinese kale transcriptome showed that the expression of CYP83a1 and CYP83b1 showed high expression level in root [24], and in plants with low contents of glucosinolates [25].
- It has been reported that the biosynthesis of glucosinolates in susceptible varieties of Chinese cabbage were active than that in resistant varieties, and glucosinolates were increased by infection of Plasmodiophora [39], while that in resistant varieties was not influenced by treatments.
- They also showed that the two susceptible varieties had different contents of in- dolic and aliphatic glucosinolates in leaves or roots, while aromatic glucosinolates were the major glucosino- lates in resistant varieties in response to Plasmodio- phora, which were differentially influenced by the addition of SA and JA [39].
- Our study was performed with 7-day old (post germination) Chinese kale seed- lings, and the whole plant contents of glucosinolates were detected.
- In summary, we performed the de novo assembly of Chinese kale genomes using SMRT sequencing com- bined with NGS sequencing.
- MeJA and BR treatments increased the content of total glucosino- lates and/or sulforaphane by upregulated the expression of genes related to glucosinolate core pathways, including CYP83b1, CYP83b1, SUR1 and/or UGT74b1.
- We concluded that the treatment of MeJA, Se and BR, especially MeJA, confer the formation and biosynthesis of glucosinolates and sulforaphane..
- High sulforaphane Chinese kale germplasm resource BOK92 (inbred line) was obtained from the seed bank of College of Horticulture and Landscape, Hunan Agricul- tural University, Changsha, China.
- solutions and treated with NaCl and 150 mM), MeJA and 120 μM), Se and 120 μM) and BR and 3.0 μM) with four triplicates of 30 seeds each.
- Extraction and measurements of sulforaphane and glucosinolate.
- The content of sulforaphane was determined using high performance liquid chromatog- raphy (HPLC: Shimadzu, Kyoto, Japan) methods with conditions as follows: C18 column (250 mm × 4.6 mm, 5 μm, at 30 °C), mobile phase (20% acetonitrile-80%.
- Glucosinolate were methanol-extracted from kale seedlings.
- 30.0 and QD <.
- Additional file 1: Figure S1.
- Additional file 2: Figure S2.
- Additional file 3: Figure S3.
- Additional file 6: Figure S4.
- Additional file 7: Figure S5.
- SXM performed the sulforaphane and glucosinolates content measure and attended data re-interpreting discussion.
- Antioxidant functions of sulforaphane: a potent inducer of phase II detoxication enzymes.
- Enhancement of sulforaphane content in cabbage outer leaves using hybrid drying technique and stepwise change of drying temperature.
- Biosynthesis and bioactivity of glucosinolates and their production in plant in vitro cultures.
- Reactivity and stability of glucosinolates and their breakdown products in foods.
- Amplification of sulforaphane content in red cabbage by pressure and temperature treatments.
- Chung FL: Disposition of glucosinolates and sulforaphane in humans after ingestion of steamed and fresh broccoli.
- Changes in Sulforaphane and Selenocysteine methyltransferase transcript levels in broccoli treated with sodium selenite.
- De novo transcriptome assembly of Chinese kale and global expression analysis of genes involved in Glucosinolate metabolism in multiple tissues.
- The chemical diversity and distribution of glucosinolates and isothiocyanates among plants.
- Variation of glucosinolates in vegetable crops of Brassica rapa.
- Metabolic engineering of aliphatic glucosinolates in Chinese cabbage plants expressing Arabidopsis MAM1, CYP79F1, and CYP83A1

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