Induction of Glucoraphasatin Biosynthesis Genes by MYB29 in Radish (Raphanus sativus L.) Roots
Abstract
:1. Introduction
2. Results
2.1. GSL Profiles in Radish Roots
2.2. RNA Sequencing and Mapping of Radish Reference Genomes
2.3. Analysis of Differentially Expressed Genes between Roots of the Radish Accessions
2.4. Identification and Expression Analysis of GSL Biosynthesis Genes in Radish Roots
2.5. GRH Content and Expression of GRH Biosynthesis Genes in Radish Roots
3. Discussion
3.1. Abundant GRH Content and Functional Possibility for Radish Breeding
3.2. Induction of GRH Biosynthesis by MYB29 Transcription Factor in Radish Roots
3.3. Expression Profiling of GRH Biosynthesis Genes in Radish Roots
4. Materials and Methods
4.1. Plant Materials
4.2. Determination of GSL Content
4.3. Total RNA Isolation and RNA-Seq Analysis
4.4. Analysis of Differentially Expressed Genes Involved in GSL Biosynthesis
4.5. Quantitative Real-Time PCR
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
GSL | Glucosinolate |
GSLs | Glucosinolates |
GRH | Glucoraphasatin |
GRA | Glucoraphanin |
GRE | Glucoraphenin |
GNA | Gluconapin |
GBN | Glucobrassicanapin |
GER | Glucoerucin |
4HGBS | 4-hydroxyglucobrassicin |
4MOGBS | 4-methoxyglucobrassicin |
NGBS | Neoglucobrassicin |
GBS | Glucobrassicin |
LGRHA | Low GRH accession |
HGRHA | High GRH accession |
GO | Gene ontology |
DEGs | Differentially expressed genes |
7MTH | 7-Methylthioheptyl |
8MTO | 8-Methylthiooctyl |
4MTB | 4-Methlthiobutyl |
3MTP | 3-Methylthiopropyl |
BCATs | Branched-chain aminotransferases |
MAMs | Methylthioalkylmalate synthases |
IPMIs | Isopropylmalate isomerases |
IPMDH | Isopropylmalate dehydrogenase |
BAT5 | Bile acid transporter 5 |
CYP | Cytochrome P450 monooxygenase |
SUR1 | S-alkyl-thiohydroximate lyase |
UGT74 | UDP-glucosyl transferase 74 |
SOTs | Sulfotransferases |
FMOGS-OXs | Flavin-monooxygenase glucosinolate S-oxygenases |
MYB | MYB transcription factor |
GRS1 | Glucoraphasatin synthase 1 |
GS-OH | 2-oxoglutarate and Fe (II)-dependent oxygenase |
IGMTs | O-methyltransferases |
AOP2 | 2-oxoglutarate-dependent dioxygenase |
qRT-PCR | Quantitative real-time Polymerase Chain Reaction |
RsActin | Raphanus sativus actin |
MeJA | Methyl jasmonate |
DS-GSL | Desulfo-glucosinolate |
DH | Doubled haploid |
FDR | False discovery rate |
ITCs | Isothiocyanates |
Wt. | Weights |
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Classification | Common Name | Abbreviation | Chemical Name | Molecular Formula |
---|---|---|---|---|
Aliphatic | Glucoraphanin | GRA | 4-(Methylsulfinyl)butyl | C12H23NO10S3 |
Glucoraphenin | GRE | 4-Methylsulfinyl-3-butenyl | C12H21NO10S3 | |
Gluconapin | GNA | 3-Butenyl | C11H19NO9S2 | |
Glucobrassicanapin | GBN | 4-Pentenyl | C12H21NO9S2 | |
Glucoerucin | GER | 4-(Methylthio)butyl | C12H23NO9S3 | |
Glucoraphasatin | GRH | 4-Methylthio-3-butenyl | C12H21NO9S3 | |
Indolic | 4-Hydroxyglucobrassicin | 4HGBS | 4-Hydroxyindol-3-ylmethyl | C16H20N2O10S2 |
4-Methoxyglucobrassicin | 4MOGBS | 4-Methoxyindol-3-ylmethyl | C17H22N2O10S2 | |
Neoglucobrassicin | NGBS | 1-Methoxyindol-3-ylmethyl | C17H22N2O10S2 | |
Glucobrassicin | GBS | 3-Indolylmethyl | C16H20N2O9S2 |
Accessions | Origin | Species | Selfing Generation | GRA | GRE | GNA | GBN | GER | GRH | NGBS | GBS | 4HGBS | 4MOGBS | Unknown | Unknown | Total GSLs |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
RA502-92 | Hungary | Raphanus sativus convar. sativus | S2 | ND | ND | 0.27 ± 0.05 | 0.04 ± 0.04 | ND | 0.17 ± 0.30 | 0.17 ± 0.04 | 0.01 ± 0.01 | 0.05 ± 0.06 | 0.08 ± 0.07 | 0.09 ± 0.15 | ND | 0.35 ± 0.05 |
IT119282-15 | Lebanon | Raphanus sativus var. sativus | S3 | 0.08 ± 0.13 | 0.12 ± 0.12 | ND | 0.66 ± 0.39 | ND | 32.70 ± 23.09 | 0.19 ± 0.05 | 0.23 ± 0.21 | 0.16 ± 0.14 | 0.01 ± 0.02 | 0.27 ± 0.22 | ND | 62.35 ± 9.14 |
IT119238-8 | Iran | Raphanus sativus var. sativus | S2 | 0.17 ± 0.29 | 0.39 ± 0.31 | ND | 0.46 ± 0.18 | ND | 47.92 ± 11.01 | 0.18 ± 0.02 | 0.20 ± 0.09 | 0.36 ± 0.23 | 0.07 ± 0.01 | 0.35 ± 0.10 | ND | 38.55 ± 5.43 |
RA280-82 | Kazakhstan | Raphanus sativus convar. sativus | S3 | 0.74 ± 0.24 | 0.32 ± 0.14 | 0.23 ± 0.05 | 0.56 ± 0.26 | ND | 39.69 ± 16.99 | 0.17 ± 0.04 | 0.38 ± 0.19 | 2.06 ± 0.58 | 0.02 ± 0.04 | 0.68 ± 0.34 | ND | 60.12 ± 6.30 |
RA157-74 | Portugal | Raphanus sativus convar. sativus | S3 | 0.85 ± 0.47 | 0.49 ± 0.37 | 0.22 ± 0.02 | 0.71 ± 0.22 | ND | 138.39 ± 31.11 | 0.19 ± 0.08 | 0.25 ± 0.11 | 1.68 ± 0.50 | 0.19 ± 0.04 | 1.51 ± 0.35 | 0.08 ± 0.13 | 122.46 ± 14.39 |
Gene ID | BLAST | |
---|---|---|
RSG02297 | nr BLAST | PREDICTED: 1-aminocyclopropane-1-carboxylate oxidase homolog 5-like, BAW81934.1| GLUCORAPHASATIN SYNTHASE 1 (Raphanus sativus) |
InterProScan | IPR026992 (Pfam); Non-haem dioxygenase N-terminal domain, | |
IPR005123 (Pfam); Oxoglutarate/iron-dependent dioxygenase | ||
Araport11 | AT1G03410.2 | 2-oxoglutarate (2OG) and Fe (II)-dependent oxygenase superfamily protein | Chr1:844782-846463 REVERSE LENGTH = 361 |
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Kang, J.-N.; Won, S.Y.; Seo, M.-S.; Lee, J.; Lee, S.M.; Kwon, S.-J.; Kim, J.S. Induction of Glucoraphasatin Biosynthesis Genes by MYB29 in Radish (Raphanus sativus L.) Roots. Int. J. Mol. Sci. 2020, 21, 5721. https://doi.org/10.3390/ijms21165721
Kang J-N, Won SY, Seo M-S, Lee J, Lee SM, Kwon S-J, Kim JS. Induction of Glucoraphasatin Biosynthesis Genes by MYB29 in Radish (Raphanus sativus L.) Roots. International Journal of Molecular Sciences. 2020; 21(16):5721. https://doi.org/10.3390/ijms21165721
Chicago/Turabian StyleKang, Ji-Nam, So Youn Won, Mi-Suk Seo, Jeongyeo Lee, Si Myung Lee, Soo-Jin Kwon, and Jung Sun Kim. 2020. "Induction of Glucoraphasatin Biosynthesis Genes by MYB29 in Radish (Raphanus sativus L.) Roots" International Journal of Molecular Sciences 21, no. 16: 5721. https://doi.org/10.3390/ijms21165721