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(A) Average height of plants (in millimeters) before harvesting for chemical analysis. (B) RNA gel blot analysis. Shown are D transcript levels in D, d x , and 35S::D plant material. No hybridization of D sequences to a D transcript was detected in d x mRNA, suggesting this as a null allele. Low and high transcript accumulation was seen in the D and 35S::D lines, respectively. 18S RNA hybridization is shown as RNA loading control.
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![Table 1 . GC-MS identification of [ 2 H6]6-OHCS and [ 2 H6]CS converted...](profile/Yuji-Kamiya/publication/13299562/figure/tbl1/AS:667825647673346@1536233450066/GC-MS-identification-of-2-H66-OHCS-and-2-H6CS-converted-from-2_Q320.jpg)
Brassinosteroids (BRs) are steroidal plant hormones essential for normal plant growth and development. Mutants in the biosynthesis or perception of BRs are usually dwarf. The tomato Dwarf gene (D), which was predicted to encode a cytochrome P450 enzyme (P450) with homology to other P450s involved in BR biosynthesis, was cloned previously. Here, we...
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Context 1
... average height of six representative individuals from d x , D, and 35S::D lines (Fig. 2A) indicates the effective comple- mentation of the d x phenotype using the 35S::D construct. Although the 35S::D line is larger than the D line, more independent 35S::D transformants need to be analyzed in detail to see whether this is a general phenomenon. Gel blot analysis (Fig. 2B) carried out on RNA extracted from d x , D, and 35S::D ...
Context 2
... of six representative individuals from d x , D, and 35S::D lines (Fig. 2A) indicates the effective comple- mentation of the d x phenotype using the 35S::D construct. Although the 35S::D line is larger than the D line, more independent 35S::D transformants need to be analyzed in detail to see whether this is a general phenomenon. Gel blot analysis (Fig. 2B) carried out on RNA extracted from d x , D, and 35S::D plant material showed that the D transcript was not detected from d x plants and was overexpressed in 35S::D plants. This is consistent with d x having a null phenotype for D, which aids the interpretation of the BR analysis and complementation ...
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The phytohormones brassinosteroid (BR), auxin, and gibberellin (GA) regulate photomorphogenesis-related hypocotyl elongation in Arabidopsis via the cooperative interaction of BZR-ARF-PIF/DELLA (BAP/D) transcription factors/regulators. Moreover, ethylene activates the PIF3 or ERF1 pathway through EIN3/EIL1 to balance hypocotyl elongation in Arabidop...
In Arabidopsis, interplay between nuclear auxin perception and trans-cellular polar auxin transport determines the transcriptional auxin response. In brevis radix (brx) mutants, this response is impaired, probably indirectly because of disturbed crosstalk between the auxin and brassinosteroid pathways. Here we provide evidence that BRX protein is p...
The plant steroid hormones, brassinosteroids (BRs), and their precursors, phytosterols, play major roles in plant growth, development, and stress tolerance. Here, we review the impressive progress made during recent years in elucidating the components of the sterol and BR metabolic and signaling pathways, and in understanding their mechanism of act...
Background
Grain size is one of key agronomic traits that determine grain yield in rice. Several regulators of grain size have been identified in rice, but the mechanisms that determine grain size and yield remain largely unknown. ResultsHere we characterize a small grain (smg11) mutant in rice, which exhibits small grains, dense panicles and the i...
Brassinosteroids (BRs) are plant polyhydroxy-steroids that play important roles in plant growth and development via extensive signal integration through direct interactions between regulatory components of different signaling pathways. Recent studies have shown that diverse helix-loop-helix/basic helix-loop-helix (HLH/bHLH) family proteins are acti...
Citations
... The deetiolated2 (det2) mutant, characterized by a dwarfed phenotype, was initially discovered in Arabidopsis [20] and subsequently found in maize [21]. Further investigation of mutants exhibiting severe pleiotropic phenotypes led to the identification of additional key BRbiosynthetic genes, such as CONSTITUTIVE PHOTOMORPHOGENESIS AND DWARFISM (CPD) [22], DWARF4 (DWF4) [23], ROTUNDFOLIA3 (ROT3) [24] and BR-6-OXIDASE1(BR6OX1) [25], in both Arabidopsis and rice. Mutants lacking these crucial BR-biosynthetic enzymes manifest various phenotypic changes, including dwarfism, delayed senescence, dark-green leaves, shortened petioles, and reduced hypocotyl length in darkness [23]. ...
Brassinosteroids (BRs), one of the major classes of phytohormones are essential for various processes of plant growth, development, and adaptations to biotic and abiotic stresses. In Arabidopsis, AtCYP90D1 acts as a bifunctional cytochrome P450 monooxygenase, catalyzing C-23 hydroxylation in the brassinolide biosynthetic pathway. The present study reports the functional characterizations of PtoCYP90D1, one of the AtCYP90D1 homologous genes from Populus tomentosa. The qRT-PCR analysis showed that PtoCYP90D1 was highly expressed in roots and old leaves. Overexpression of PtoCYP90D1 (PtoCYP90D1-OE) in poplar promoted growth and biomass yield, as well as increased xylem area and cell layers. Transgenic plants exhibited a significant increase in plant height and stem diameter as compared to the wild type. In contrast, the CRISPR/Cas9-generated mutation of PtoCYP90D1 (PtoCYP90D1-KO) resulted in significantly decreased biomass production in transgenic plants. Further studies revealed that cell wall components increased significantly in PtoCYP90D1-OE lines but not in PtoCYP90D1-KO lines, as compared to wild-type plants. Overall, the findings indicate a positive role of PtoCYP90D1 in improving growth rate and elevating biomass production in poplar, which will have positive implications for its versatile industrial or agricultural applications.
... As a result, plant mutants that are defective in the biosynthetic steps leading from mevalonic acid to sterol, as well as in the steps involved in modifying sterols to brassinolide, often display the characteristic dwarf phenotype [41,42] . DWARF1 was reported as a biosynthetic enzyme involved in the conversion of the early brassinosteroid precursor 24-methylenecholesterol to campesterol [43] . In DWARF1, a conserved flavin adenine dinucleotide (FAD)-binding domain has been observed across various plant species [11,44,45] . ...
Curd setting height, a critical component of plant height, plays an important role in ideotype construction and yield improvement in cauliflower. However, the underlying mechanisms governing this trait in Brassica crops remain unclear. In this study, we developed two F2 populations by crossing a cauliflower DH line, ZAASC4101, having a short main stem, with two Brassica oleracea inbred lines possessing a long main stem. Subsequently, two high-density linkage maps were constructed for each F2 population using a 10K MNP (multiple nucleotide polymorphism) array developed for Brassica oleracea species. These maps covered genetic distances of 8,373.66 and 8,250.01 cM, respectively. Notably, a major QTL related to curd setting height (BocDWARF1) was consistently identified on chromosome C01 with the highest LOD value in both F2 populations. Through the analysis of the recombinants identified from the F2:3 and F3:4 populations at the BocDWARF1 locus, we successfully fine-mapped this locus to a narrow interval delimited by SNP markers 1_40298271 and 1_40400283. This interval corresponds to an approximately 102-kb region harboring 12 predicted genes. Combined with parental resequencing data with HDEM reference genome annotation information, we proposed BolC1t04399H as the most promising candidate gene. This gene exhibits homology to the well-known plant height-controlling gene DWARF1 and shows a total of four nonsynonymous SNPs in the exon region between the parents. This study lays the foundation for map-based cloning of BocDWARF1 and provides valuable insights for breeding improved cauliflower varieties with the ideal curd setting height.
... To date, a number of mutants related to BR biosynthesis or perception, such as cpd, dwf1, dwf4, dwf7/ste1, det2, bri1 and brd1 have been isolated in Arabidopsis (Choe et al. 1998(Choe et al. , 1999Chory et al. 1991;Clouse et al. 1996;Klahre et al. 1998;Mori et al. 2002;Tanabe et al. 2005;Hong et al. 2005;Szekeres et al. 1996;Wang et al. 2017a). It has been demonstrated that mutants blocking various steps of BR biosynthesis or signal transduction generally result in characteristic dwarf phenotypes, which include extremely dwarfed statures, prolonged life span, delayed flowering and senescence, dark-green and small curled leaves, reduced fertility and altered photomorphogenesis in dark conditions (Bishop et al. 1999;Mori et al. 2002;Kwon and Choe 2005;Li et al. 2013;Nolan et al. 2020). Although BR-deficient mutants and signaling mutants are morphologically alike, BR-deficient mutants can be rescued by exogenous application of brassinolide (BL), while BR-signaling mutants often exhibit insensitivity to BL treatment (Nolan et al. 2020). ...
Key message
A novel super compact mutant, scp-3, was identified using map-based cloning in cucumber. The CsDWF7 gene encoding a delta7 sterol C-5(6) desaturase was the candidate gene of scp-3.
Abstract
Mining dwarf genes is important in understanding stem growth in crops. However, only a small number of dwarf genes have been cloned or characterized. Here, we characterized a cucumber (Cucumis sativus L.) dwarf mutant, super compact 3 (scp-3), which displays shortened internodes and dark green leaves with a wrinkled appearance. The photosynthetic rate of scp-3 is significantly lower than that of the wild type. The dwarf phenotype of scp-3 mutant can be partially rescued by the exogenous brassinolide (BL) application, and the endogenous brassinosteroids (BRs) levels in the scp-3 mutant were significantly lower compared to the wild type. Microscopic examination revealed that the reduced internode length in scp-3 resulted from a decrease in cell size. Genetic analysis showed that the dwarf phenotype of scp-3 was controlled by a single recessive gene. Combined with bulked segregant analysis and map-based cloning strategy, we delimited scp-3 locus into an 82.5 kb region harboring five putative genes, but only one non-synonymous mutation (A to T) was discovered between the mutant and its wild type in this region. This mutation occurred within the second exon of the CsGy4G017510 gene, leading to an amino acid alteration from Leu¹⁵⁶ to His¹⁵⁶. This gene encodes the CsDWF7 protein, an analog of the Arabidopsis DWF7 protein, which is known to be involved in the biosynthesis of BRs. The CsDWF7 protein was targeted to the cell membrane. In comparison to the wild type, scp-3 exhibited reduced CsDWF7 expression in different tissues. These findings imply that CsDWF7 is essential for both BR biosynthesis as well as growth and development of cucumber plants.
... Then, BRI1 binds to BR to form a BRI1-BR complex that is capable of forming an intercalative interface outside the cell to induce binding of the extracellular domain of the co-receptor BAK1 to form a BRI1-BR-BAK1 complex. Mutual phosphorylation between the kinase regions of BRI1 and BAK1 occurs so that BRI1 is fully activated (Bishop et al., 1999). As a result, BR signaling can continue. ...
Brassinolides (BRs) are a kind of endogenous plant hormones that plays an important role in regulating fruit ripening. BRs can regulate the process of fruit ripening by interacting with plant growth regulators and transcription factors. In this review, we have discussed recent research advances on the role of BRs in fruit ripening and its relation to plant growth regulators and transcription factors. In addition, BRs signaling molecules which are involved in fruit ripening and ripening-related genes were also highlighted. Consequently, the review can provide a better theoretical basis for improving fruit quality and regulating fruit ripening to speed up market availability, and for studying the mechanisms of BRs in fruit ripening.
... For example, d x , a tomato BR-deficient mutant with an impaired Dwarf gene encoding a BR C-6 oxidase, exhibits severe dwarfism. d x fruits show delayed ripening and reduced yield and dry mass, which can be partially rescued by BR application (Bishop et al., 1996(Bishop et al., , 1999Nomura et al., 2005;Lisso et al., 2006). Third, intense BR biosynthesis occurs during tomato fruit development, suggesting that BRs are required for tomato fruit development (Montoya et al., 2005;Lisso et al., 2006). ...
The plant hormone ethylene is essential for climacteric fruit ripening, although it is unclear how other phytohormones and their interactions with ethylene might affect fruit ripening. Here, we explored how brassinosteroids (BRs) regulate fruit ripening in tomato (Solanum lycopersicum) and how they interact with ethylene. Exogenous BR treatment and increased endogenous BR contents in tomato plants overexpressing the BR biosynthetic gene SlCYP90B3 promoted ethylene production and fruit ripening. Genetic analysis indicated that the BR signaling regulators Brassinazole‐resistant1 (SlBZR1) and BRI1‐EMS‐suppressor1 (SlBES1) act redundantly in fruit softening. Knocking out SlBZR1 inhibited ripening through transcriptome reprogramming at the onset of ripening. Combined transcriptome deep sequencing and chromatin immunoprecipitation followed by sequencing identified 73 SlBZR1‐repressed targets and 203 SlBZR1‐induced targets involving major ripening‐related genes, suggesting that SlBZR1 positively regulates tomato fruit ripening. SlBZR1 directly targeted several ethylene and carotenoid biosynthetic genes to contribute to the ethylene burst and carotenoid accumulation to ensure normal ripening and quality formation. Furthermore, knock‐out of Brassinosteroid‐insensitive2 (SlBIN2), a negative regulator of BR signaling upstream of SlBZR1, promoted fruit ripening and carotenoid accumulation. Taken together, our results highlight the role of SlBZR1 as a master regulator of tomato fruit ripening with potential for tomato quality improvement and carotenoid biofortification.
... DET2-encoded reductase participates in reducing 22 of 23di OH-4-EN-3-One to 6-deoxO3DT (3-dihydro-6-deoxoteasterone) to accelerate the BR biosynthesis (Poppenberger et al., 2011). DWARF, another BRs biosynthetic enzyme, encodes CYP80 to mediate c-6 oxidation of 6-deoxybrassinosterol (CR) to testosterone (CN) in tomatoes and participates in the synthesis of BRs (Bishop et al., 1999). BRI1 can activate the signaling of BRs. ...
Yeast cell walls (YCW) are promising bio-based elicitors for controlling post-harvest fruit decay. In this study, 1% YCW induction increased the resistance of cherry tomato fruits, reducing disease incidence by 66%. This study aimed to explore the interaction of hormones and crosstalk with MAPKs (mitogen-activated protein kinases) in the early response of resistance regulation in cherry tomato fruits treated with YCW and U0126. We analyzed the temporal changes in hormone content, the expression of critical genes involved in phytohormone biosynthesis, and signal transduction in cherry tomato fruits response to the induction. Results revealed that jasmonic acid (JA) and brassinosteroids (BR) significantly regulated early resistance response in fruit induced by 1% YCW. The salicylic acid (SA) pathway is inhibited by the activation of the JA pathway. JA and SA signaling pathway crosstalk with the MAPK3 pathway. BR plays an essential role in the regulation of fruit resistance. The BR pathway may function independently when JA/SA and MAPK3 pathways are inhibited.
... There are five enzymes that play key roles in the BR synthesis pathway, which are steroid 22 α-hydroxylase (90B/724B) [26,27], steroid 5 α-reductase (Det2) [28,29], C-3 oxidase (90A1) [30], C-23 hydroxylase (90C1/D1) [31], and C-6 oxidase (85A1/2) [32]. These key enzymes in the synthesis process not only participate in the reaction in one step, but also play a role in a variety of pathways. ...
Pak choi needs to be exposed to low temperature (vernalized) before flowering will initiate. Early bolting caused by low temperature often occurs in spring and leads to significant economic losses. Therefore, it is of great practical significance to study the flowering process of pak choi. Brassinolide (BR) plays a certain role in the flowering process of pak choi. In order to better understand the role of BR in the flowering process of pak choi, the BR content in the shoot apices of pak choi at different growth stages was determined by enzyme-linked immunosorbent assay (ELISA). The results showed that the BR content increased after low-temperature treatment. With the progress of vegetative growth, the BR content decreased and reached the lowest at 10 days after transplanting (V10), then the content increased and reached a small peak at the critical period of floral bud differentiation (S0). After initiation of floral bud differentiation, the content decreased at floral bud differentiation stage 1 (S1), and then gradually increased and reached a peak at floral bud differentiation stage 3 (S3). In order to clarify the molecular mechanism of BR content changes, we analyzed the expressions of key enzymes coding genes in the BR metabolic pathway, and found that six major synthase-encoding genes (Bra008760, Bra030023, Bra036097, Bra027405, Bra011678, and Bra025409) were upregulated at the critical period of floral bud differentiation, leading to the increase in BR content, which were consistent with changes in the BR content. By analyzing the functions of differentially expressed genes (DEGs) between the vegetative growth stage (V10) and the critical period of floral bud differentiation (S0), 21 DEGs were found to be related to BR metabolism. These findings can provide a reference for elucidating the molecular mechanism of BR regulating the flowering process of pak choi.
... SELF-PRUNING encodes a 23 kDa CETS (CEN-TRORADIALIS/TERMINAL FLOWER 1/SELF-PRUNING) family modulator that determines the potential of the shoot apical meristem for continuous growth (Pnueli et al., 2001). The DWARF gene encodes a P450 protein that catalyzes the C-6 oxidation of 6-deoxocastasterone to castasterone in brassinosteroid biosynthesis (Bishop et al., 1999). Furthermore, Micro-Tom tomato has an additional mutation that decreases the internode length without affecting the active gibberellin levels (Martí et al., 2006). ...
The target of rapamycin complex (TORC) plays a key role in plant cell growth and survival by regulating the gene expression and metabolism according to environmental information. TORC activates transcription, mRNA translation, and anabolic processes under favorable conditions, thereby promoting plant growth and development. Tomato fruit ripening is a complex developmental process promoted by ethylene and specific transcription factors. TORC is known to modulate leaf senescence in tomato. In this study, we investigated the function of TORC in tomato fruit ripening using virus-induced gene silencing (VIGS) of the TORC genes, TOR, lethal with SEC13 protein 8 (LST8), and regulatory-associated protein of TOR (RAPTOR). Quantitative reverse transcription-polymerase chain reaction showed that the expression levels of tomato TORC genes were the highest in the orange stage during fruit development in Micro-Tom tomato. VIGS of these TORC genes using stage 2 tomato accelerated fruit ripening with premature orange/red coloring and decreased fruit growth, when control tobacco rattle virus 2 (TRV2)-myc fruits reached the mature green stage. TORC-deficient fruits showed early accumulation of carotenoid lycopene and reduced cellulose deposition in pericarp cell walls. The early ripening fruits had higher levels of transcripts related to fruit ripening transcription factors, ethylene biosynthesis, carotenoid synthesis, and cell wall modification. Finally, the early ripening phenotype in Micro-Tom tomato was reproduced in the commercial cultivar Moneymaker tomato by VIGS of the TORC genes. Collectively, these results demonstrate that TORC plays an important role in tomato fruit ripening by modulating the transcription of various ripening-related genes.
... In tomato, multiple genes essential for GA synthesis are functionally characterized (Carvalho et al., 2011;Illouz-Eliaz et al., 2019;Koornneef et al., 1990;Li et al., 2012). Similarly, this phenotype can be observed in tomatoes with different hormonal mutations, such as mutations in genes involved in brassinosteroid (Bishop et al., 1999;Li et al., 2016;Martí et al., 2006;Nie et al., 2017) or indole-3-acetic acid syntheses (Higashide et al., 2014;Leyser, 2018;Zhao, 2018). However, such mutation-derived phenotypes with reduced height have not been utilized commercially, particularly in large-fruited fresh-market tomato varieties, often due to extreme reductions in the fruit size (mediumsized or larger fruits are classified as typical marketable fruits in largefruited fresh-market tomato in the United States; Florida Tomato Committee, 2022) and marketable yield, which were observed for tomato germplasm releases and varieties (Scott & Harbaugh, 1989; J.W. Scott and R.G. Gardner, personal communications; a comparison of fruit size shown in Figure S1), and experimental tomato lines (Higashide et al., 2014;Liu et al., 2018;Srivastava & Handa, 2005) that harbor mutant allele(s). ...
Reduced plant height due to shortened stems is beneficial for improving crop yield potential, better resilience to biotic/abiotic stresses, and rapid crop producer adoption of the agronomic and management practices. Breeding tomato plants with a reduced height, however, poses a particular challenge because this trait is often associated with a significant fruit size (weight) reduction. The tomato BRACHYTIC (BR) locus controls plant height. Genetic mapping and genome assembly revealed three flowering promoting factor 1 (FPF1) genes located within the BR mapping interval, and a complete coding sequence deletion of the telomere proximal FPF1 (Solyc01g066980) was found in the br allele but not in BR. The knock-out of Solyc01g066980 in BR large-fruited fresh-market tomato reduced the height and fruit yield, but the ability to produce large size fruits was retained. However, concurrent yield evaluation of a pair of sister lines with or without the br allele revealed that artificial selection contributes to commercially acceptable yield potential in br tomatoes. A network analysis of gene-expression patterns across genotypes, tissues, and the gibberellic acid (GA) treatment revealed that member(s) of the FPF1 family may play a role in the suppression of the GA biosynthesis in roots and provided a framework for identifying the responsible molecular signaling pathways in br-mediated phenotypic changes. Lastly, mutations of br homologs also resulted in reduced height. These results shed light on the genetic and physiological mechanisms by which the br allele alters tomato architecture.
... Classical dwarf mutant nana plant 1 with feminized male flowers in maize was reported to carry a loss-offunction in a DET2 homolog gene in the BR biosynthesis pathway (Hartwig et al. 2011). The block in C-6 oxidation of BRs results in the dwarf phenotype of tomato d x mutant (Bishop et al. 1999). In cucumber, several genes underlying BR related mutants scp-1, scp-2, cpa have been reported with the typical phenotype of BR-deficient mutant including extremely dwarf, no lateral branches, smaller wrinkled and dark green leaves, and lower fertility Hou et al. 2017;Zhang et al. 2021). ...
Key message
A novel compact plant architecture mutant, cpa-2, was identified from EMS-induced mutagenesis. Bulked segregant analysis sequencing and map-based cloning revealed CsDWF1 encoding C-24 reductase enzyme as the candidate gene.
Abstract
The compact architecture is a vital and valuable agronomic trait that helps to reduce the labor of plant management, and improve the fruit yield by increasing planting density in cucumbers. However, the molecular basis underlying the regulation of plant architecture in cucumber is complex and largely unknown. In this study, a novel recessive compact allele, designated as cpa-2 (compact plant architecture-2) was fine mapped in a 109 kb region on chromosome 7 by the strategy of bulked segregant analysis sequencing combined with map-based cloning. Gene annotation of the corresponding region revealed that the CsaV3_7G030530 (CsDWF1) gene encoding C-24 reductase, which acts as the key enzyme in brassinosteroids biosynthesis, functions as the candidate gene for cpa-2. Sequence analysis showed that a single-nucleotide mutation (G to A) in the second exon of CsaV3_7G030530 caused an amino acid substitution from E⁵⁰² to K⁵⁰². Compared with wild-type CCMC, CsDWF1 had lower expression levels in the stem, leaf and ovary of cpa-2. In addition, the compact phenotype in cpa-2 could be partially restored by exogenous BR application. Transcriptome analysis revealed that many genes related to plant growth hormones were differentially expressed in cpa-2 plants. This is the first report about the characterization and cloning of the CsDWF1 gene. This work revealed the importance of CsDWF1 in plant development regulation and extended our understanding of the interaction between BRs and other hormones for plant architecture development.
