Brassinolide Research Articles

Structural insights into brassinosteroid export mediated by the Arabidopsis ABC transporter ABCB1

Brassinosteroids (BRs) are steroidal phytohormones indispensable for plant growth, development, and responses to environmental stresses. The export of bioactive BRs to the apoplast is essential for BR signalling initiation, which requires binding of BR molecule to the extracellular domains of the plasma membrane-localized receptor complex. We have previously shown that the Arabidopsis thaliana ATP-binding cassette (ABC) transporter, ABCB19, functions as a BR exporter, and together with its close homologue, ABCB1, positively regulate BR signalling. Here, we demonstrate that ABCB1 is another BR transporter. The ATP hydrolysis activity of ABCB1 was stimulated by bioactive BRs, and its transport activity was confirmed in proteoliposomes and protoplasts. Structures of ABCB1 in substrate-unbound (apo), brassinolide (BL)-bound, and ATP plus BL-bound states were determined. In the BL-bound structure, BL was bound to the hydrophobic cavity formed by the transmembrane domain, and triggered local conformational changes. Together, our data provide additional insights into the ABC transporter-mediated BR export.

Exogenous Brassinolide Ameliorates the Adverse Effects of Gamma Radiation Stress and Increases the Survival Rate of Rice Seedlings by Modulating Antioxidant Metabolism.

Gamma irradiation-based mutant creation is one of the most important methods for rice plant mutagenesis breeding and molecular biology research. Although median lethal dose irradiation severely damages rice seedlings, applying brassinolide (BR) can increase the survival rate of irradiated seedlings. In this study, we investigated the effects of soaking seeds in solutions containing different BR concentrations (0.001, 0.01, 0.1, 1.0, and 5.0 μmol/L) and then spraying the resulting seedlings twice with 0.1 μmol/L BR. The combined BR treatments markedly decreased the superoxide anion (O2•-), hydrogen peroxide (H2O2), and malondialdehyde contents but increased the chlorophyll content. An appropriate BR treatment of gamma-irradiated samples substantially increased the activities of the antioxidant enzymes superoxide dismutase, peroxidase, and ascorbate peroxidase as well as the proline, ascorbic acid, and glutathione contents in rice seedling shoots. The BR treatment also promoted the growth of seedlings derived from irradiated seeds and increased the shoot and root fresh and dry weights. Most notably, soaking seeds in 0.01 or 0.1 μmol/L BR solutions and then spraying seedlings twice with 0.1 μmol/L BR significantly increased the final seedling survival rate and decreased mutant loss. The study results suggest that exogenous BR treatments can protect rice seedlings from gamma irradiation stress by enhancing antioxidant metabolism.

Characterization of a Rho GDP dissociation inhibitor gene OsRhoGDI1 from rice

The Rho GDP dissociation inhibitor gene OsRhoGDI1 was isolated as the putative partner of OsRac5 from rice young panicles by the yeast two-hybrid screening, but the characterization and functions of the gene remains unknown. In this study, on the basis of the bioinformatics analysis, expression profile chip mining and RT-qPCR were employed to analyze the expression pattern of OsRhoGDI1, and it was found that the gene was widely expressed in various tissues of rice, especially with high abundance during the development of young panicles, and its expression was regulated by abscisic acid (ABA), brassinolide (BL), salicylic acid (SA), indole-3-acetic acid (IAA), gibberellin (GA) and high salinity. Subcellular localization identification showed that OsRhoGDI1 was distributed in the cytomembrane, cytoplasm, and nucleus. Bimolecular fluorescence completion (BiFC) was further performed to identify the in vivo interaction between OsRhoGDI1 and OsRac5, the results suggested that the interaction occurred in the nucleus, cytomembrane, and cytoplasm. In addition, OsRhoGDI1 interacted with OsRac1 in the cytomembrane. Our results indicated that OsRhoGDI1 may be involved in rice panicle development, and participate in regulating various biological processes such as rice growth and development, hormone response, and abiotic stress by regulating the activities of different Rho/Rop proteins via interactions.

The HAT1 transcription factor regulates photomorphogenesis and skotomorphogenesis via phytohormone levels.

Plants dynamically modulate their growth and development to acclimate to the fluctuating light environment via a complex phytohormone network. However, the dynamic molecular regulatory mechanisms underlying how plants regulate phytohormones during skotomorphogenesis and photomorphogenesis are largely unknown. Here, we identified a HD-ZIP II transcription factor, HOMEODOMAIN ARABIDOPSIS THALIANA1 (HAT1), as a key node that modulates the dose effects of brassinosteroids (BR) and auxin on hypocotyl growth during skotomorphogenesis and photomorphogenesis. Compared with the wild-type (Col-0), both HAT1 loss of function and its overexpression led to disrupted photomorphogenic and skotomorphogenic hypocotyl growth. HAT1 overexpression (HAT1OX) plants displayed longer hypocotyls in the light but shorter hypocotyls in darkness, whereas the triple mutant hat1hat2hat3 showed the opposite phenotype. Furthermore, we found that CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) interacted with dephosphorylated HAT1 and facilitated the degradation of HAT1 by ubiquitination in darkness, while HAT1 was phosphorylated and stabilized by BRASSINOSTEROID INSENSITIVE2 (BIN2) in the light. Interestingly, we observed distinct dose-dependent effects of BR and auxin on hypocotyl elongation under varying light conditions and that HAT1 functioned as a key node in this process. The shorter hypocotyl of HAT1OX in darkness was due to the inhibition of BR biosynthetic gene BRASSINOSTEROID-6-OXIDASE2 (BR6OX2) expression to reduce BRs content, while brassinolide (BL) treatment alleviated this growth repression. In the light, HAT1 inhibited BR biosynthesis but enhanced auxin signaling by directly repressing IAA3/SHORT HYPOCOTYL 2 (SHY2) expression. Our findings uncover a dual function of HAT1 in regulating BR biosynthesis and auxin signaling that is crucial for ensuring proper skotomorphogenic and photomorphogenic growth.

Synergistic variation in abscisic acid and brassinolide treatment signaling component alleviates fruit quality of ‘Shine Muscat’ grape during cold storage

Grape fruit are harvested in the late summer or early fall and need to be stored at low temperatures to prevent enfeeblement and prolong their shelf-life. This study aimed to determine the effects of abscisic acid (ABA), brassinolide (BR) and ABA + BR (ABR) treatment on the berry quality of ‘Shine Muscat’ under low temperatures. ABA and BR maintained fruit appearance, cellular structure, weight, firmness. ABR treatments reduced the loss of fruit aroma. Furthermore, the transcriptome and metabolome analysis revealed that ABA, BR, and ABR treatments maintained the quality of fruits during the low temperatures period by influencing chlorophyll metabolism, carotenoid metabolism, flavonoid metabolism, unsaturated fatty acid, and terpene metabolism. These findings identify key genes and metabolites for ABA and BR-induced maintenance of grape fruit quality during cold storage, expanding our understanding of postharvest storage quality maintenance of grape fruit at the transcript and metabolic levels.

The effect of several growth regulators and biostimulant on biochemical and physiological changes in acclimation of micropropagated Echinacea purpurea Moench. ‘Raspberry Truffle’

Micropropagation is currently one of the primary methods for plant propagation, known for its efficiency in producing disease-free and vigorous plants. However, the final stage of this—acclimatization, is critical due to the transfer from a controlled in vitro environment to an external one. To reduce mortality and alleviate acclimation stress, plant growth regulators (PGRs) or biostimulants can be employed. This study investigated the effects of exogenously sprayed PGRs: 0.001 mg L−1 abscisic acid (ABA), 0.001 mL L−1 brassinolide (BL), 0.001 mL L−1 24-epibrassinolide (24-epiBL), and 0.3 mL L−1 biostimulant Goteo on the physiological and biochemical responses of Echinacea purpurea ‘Raspberry Truffle’ plantlets during the acclimation process. The effects of treatments at various acclimation stages on chlorophyll (chl) and carotenoids content, hydrogen peroxide (H2O2), catalase (CAT), malondialdehyde (MDA), free amino acids, soluble proteins, total soluble sugar and reduced soluble sugars were tested in this research. The results confirmed changes in biochemical parameters, including an increase in chlorophyll and carotenoids in the acclimatization period where the highest level obtained by BL spraying. A decline in stomatal conductance was also observed, where ABA influenced the most on drop. It was also recorded the decrease in H2O2 and MDA concentration. CAT activity increased, especially with biostimulant treatment. We recorded an increase in total soluble proteins along acclimatization. Goteo affected the most on morphology parameters, ABA, BL and 24-epiBL increased acclimatization efficiency. Our studies indicate that potentially the most effective substances in the acclimatization of E. purpurea ‘Raspberry Truffle’ are brassinosteroids and ABA.

Interactions between Brassinosteroids and Strigolactones in Alleviating Salt Stress in Maize.

Exogenous brassinolide (BR) and strigolactones (SLs) play an important role in alleviating salt stress in maize. We studied the morphological and physiological responses of the salt-sensitive genotype PH4CV and salt-tolerant genotype Zheng58 to BR (1.65 nM), SL (1 µM), and BS (1.65 nM BR + 1 µM SL) under salt stress. Phenotypic analysis showed that salt stress significantly inhibited the growth of maize seedlings and significantly increased the content of Na+ in the roots. Exogenous hormones increased oxidase activity and decreased Na+ content in the roots and mitigated salt stress. Transcriptome analysis showed that the interaction of BR and SL is involved in photosynthesis-antenna proteins, the TCA cycle, and plant hormone signal transduction pathways. This interaction influences the expression of chlorophyll a/b-binding protein and glucose-6-phosphate isomerase 1 chloroplastic, and aconitase genes are affected. Furthermore, the application of exogenous hormones regulates the expression of genes associated with the signaling pathways of cytokinin (CK), gibberellins (GA), auxin (IAA), brassinosteroid (BR), abscisic acid (ABA), and jasmonic acid (JA). Additionally, exogenous hormones inhibit the expression of the AKT2/3 genes, which are responsible for regulating ion transduction and potassium ion influx. Four candidate genes that may regulate the seedling length of maize were screened out through WGCNA. Respective KOG notes concerned inorganic ion transport and metabolism, signal transduction mechanisms, energy production and conversion, and amino acid transport and metabolism. The findings of this study provide a foundation for the proposition that BR and SL can be employed to regulate salt stress alleviation in maize.

Combined application of silica nanoparticles and brassinolide promoted the growth of sugar beets under deficit irrigation

Silica nanoparticles (SiNPs) and brassinolide (BR) have been used as nano-fertilizer and growth regulator, respectively to enhance crop tolerance to abiotic stress. However, it is unclear whether a combination of the two (BR+SiNPs) is more beneficial than single application of BR or SiNPs to improve the growth of deficit-irrigated sugar beets. In this study, a two-year (2022-2023) field experiment was conducted to investigate the effects of foliar spraying of water (CK), SiNPs, BR, and BR+SiNPs on the antioxidant defense, photosynthetic capacity, dry matter accumulation, nutrient uptake, and yield of sugar beets under full irrigation (100% of crop evapotranspiration (ETc), W1) and deficit irrigation (60% ETc, W2). The results showed that compared with the application of BR or SiNPs, the application of BR+SiNPs could enhance the antioxidant defense, osmoregulation, and photosynthesis of the full-irrigated and deficit-irrigated sugar beet leaves, and ultimately improved the water status, growth, and yield of sugar beet plants. There was no significant difference in the net revenue (NR) between BR+SiNPs treatment and CK under W1 conditions. However, the NR of the BR+SiNPs treatment increased by 27.0% (p < 0.05) compared with that of CK under W2 conditions, and there was no significant difference in NR between BR+SiNPs and SiNPs treatments. A comprehensive evaluation using entropy weight combined with technique for order preference by similarity to ideal solution method found that under deficit irrigation condition, spraying SiNPs could improve the growth of sugar beet, increase the TY, NR, and water use efficiency, and reduce costs compared with spraying BR+SiNPs. Therefore, foliar spraying of SiNP on deficit-irrigated sugar beets can be used to improve sugar beet growth and reduce the potential economic losses caused by deficit irrigation.

Functional Analysis of the PoSERK-Interacting Protein PorbcL in the Embryogenic Callus Formation of Tree Peony (Paeonia ostii T. Hong et J. X. Zhang).

SERK is a marker gene for early somatic embryogenesis. We screened and functionally verified a SERK-interacting protein to gain insights into tree-peony somatic embryogenesis. Using PoSERK as bait, we identified PorbcL (i.e., the large subunit of Rubisco) as a SERK-interacting protein from a yeast two-hybrid (Y2H) library of cDNA from developing tree-peony somatic embryos. The interaction between PorbcL and PoSERK was verified by Y2H and bimolecular fluorescence complementation analyses. PorbcL encodes a 586-amino-acid acidic non-secreted hydrophobic non-transmembrane protein that is mainly localized in the chloroplast and plasma membrane. PorbcL was highly expressed in tree-peony roots and flowers and was up-regulated during zygotic embryo development. PorbcL overexpression caused the up-regulation of PoSERK (encoding somatic embryogenesis receptor-like kinase), PoAGL15 (encoding agamous-like 15), and PoGPT1 (encoding glucose-6-phosphate translocator), while it caused the down-regulation of PoLEC1 (encoding leafy cotyledon 1) in tree-peony callus. PorbcL overexpression led to increased indole-3-acetic acid (IAA) content but decreasing contents of abscisic acid (ABA) and 6-benzyladenosine (BAPR). The changes in gene expression, high IAA levels, and increased ratio of IAA to ABA, BAPR, 1-Aminocyclopropanecarboxylic acid (ACC), 5-Deoxystrigol (5DS), and brassinolide (BL) promoted embryogenesis. These results provide a foundation for establishing a tree-peony embryogenic callus system.

Comparative analysis of element and hormone content in zygotic embryos of Pinus elliottii and P. elliottii × P. caribaea

Somatic embryogenesis is a crucial method for achieving clonal forestry in conifers. Understanding the development of zygotic embryos is essential not only for enhancing the efficiency and quality of somatic embryogenesis, but also for advancing forestry breeding programs. This study investigated dynamic changes of element and hormone contents during ZE development of Pinus elliottii and its hybrid P. elliottii × P. caribaea. Significant differences in embryo development speed among different clones were observed. Elemental analysis was conducted using inductively coupled plasma mass spectrometry (ICP-MS) and identified 68 elements, including major, minor, and beneficial elements. In both species, the contents of potassium (K), calcium (Ca), iron (Fe), boron (B) and five beneficial elements decreased during early ZE development, while phosphorus (P) and copper (Cu) increased. Significantly higher levels of K, Ca and Fe at the initial stage, and sulfur (S) and nickel (Ni) decreased at later stages were detected in P. elliottii than in the hybrid. For the other elements, except for very few significant differences at certain stages, most differences between the two species did not reach a significant level. The contents of endogenous hormones were determined and different accumulation patterns were detected in most hormones between the two species, except abscisic acid (ABA) which simultaneously decreased with developments by stage 8. Significant differences were found in indole-3-acetic acid (IAA) contents at most stages between species, while higher levels of total cytokinin (CK) at each stage were detected in the hybrid in comparison with those in P. elliottii. As a result, lower IAA to CK ratios in the hybrid than in P. elliottii. Methyl jasmonate (JA-me) and gibberellin A3 (GA3) contents showed a similar pattern and exhibited an M-shaped fluctuation in the hybrid. Furthermore, JA-me, GA3, gibberellin A4 (GA4) and brassinolide (BR) showed significantly higher levels in the hybrid than in P. elliottii. K-means clustering and correlation analyses were used to explore relationships between elements and hormones during embryo development, revealing complex interplay in both species. These data indicate different requirement in element and hormone contents for embryogenesis and suggest species-specific media composition for each step in somatic embryogenesis. The findings provide insights into their developmental processes and informing future research and applications in somatic embryogenesis and forestry breeding.

Physiological and Proteomic Analysis of Various Priming on Rice Seed under Chilling Stress.

Rice (Oryza sativa L.) cultivation using direct seeding is susceptible to chilling stress, particularly during seed germination and early seedling growth in the early season of a double cropping system. Alternatively, seed priming with various plant growth-promoting hormones is an effective technique to promote rapid and uniform emergence under chilling stress. Therefore, we evaluated the impact of gibberellin A3 (GA3) and brassinolide (BR) priming on rice seed emergence, examining their proteomic responses under low-temperature conditions. Results indicated that GA3 and BR increased the seed germination rate by 22.67% and 7.33% at 72 h and 35% and 15% at 96 h compared to the control (CK), respectively. Furthermore, proteomic analysis identified 2551, 2614, and 2592 differentially expressed proteins (DEPs) in GA, BR, and CK, respectively. Among them, GA exhibited 84 upregulated and 260 downregulated DEPs, while BR showed 112 upregulated and 102 downregulated DEPs, and CK had 123 upregulated and 81 downregulated DEPs. Notably, under chilling stress, both GA3 and BR are involved in peroxide metabolism, phenylpropanoid biosynthesis, and inositol phosphate metabolism, enhancing antioxidant capacity and providing energy substances for germination. In addition, GA3 triggers the specific regulation of stress responsive protein activation, GTP activation, and ascorbic acid biosynthesis and promotes the stability and integrity of cell membranes, as well as the synthesis of cell walls, providing physical defense for seeds to resist low temperatures. At the same time, BR triggers specific involvement in ribosome synthesis and amino acid synthesis, promoting biosynthetic ability and metabolic regulation to maintain plant life activities under low-temperature stress. Furthermore, the various genes' expression (OsJ_16716, OsPAL1, RINO1) confirmed GA3 and BR involved in peroxide metabolism, phenylpropanoid biosynthesis, and inositol phosphate metabolism, enhancing antioxidant capacity and providing energy substances for germination. This study provides valuable insights into how rice seed embryo responds to and tolerates chilling stress with GA3 seed priming.

Transcriptomic and Physiological Studies Unveil that Brassinolide Maintains the Balance of Maize's Multiple Metabolisms under Low-Temperature Stress.

Low-temperature (LT) is one of the major abiotic stresses that restrict the growth and development of maize seedlings. Brassinolides (BRs) have been shown to enhance LT tolerance in several plant species; the physiological and molecular mechanisms by which BRs enhance maize tolerance are still unclear. Here, we characterized changes in the physiology and transcriptome of N192 and Ji853 seedlings at the three-leaf stage with or without 2 μM 2,4-epibrassinolide (EBR) application at 25 and 15 °C environments via high-performance liquid chromatography and RNA-Sequencing. Physiological analyses revealed that EBR increased the antioxidant enzyme activities, enhanced the cell membrane stability, decreased the malondialdehyde formation, and inhibited the reactive oxygen species (ROS) accumulation in maize seedlings under 15 °C stress; meanwhile, EBR also maintained hormone balance by increasing indole-3-acetic acid and gibberellin 3 contents and decreasing the abscisic acid level under stress. Transcriptome analysis revealed 332 differentially expressed genes (DEGs) enriched in ROS homeostasis, plant hormone signal transduction, and the mitogen-activated protein kinase (MAPK) cascade. These DEGs exhibited synergistic and antagonistic interactions, forming a complex LT tolerance network in maize. Additionally, weighted gene co-expression network analysis (WGCNA) revealed that 109 hub genes involved in LT stress regulation pathways were discovered from the four modules with the highest correlation with target traits. In conclusion, our findings provide new insights into the molecular mechanisms of exogenous BRs in enhancing LT tolerance of maize at the seedling stage, thus opening up possibilities for a breeding program of maize tolerance to LT stress.

A brassinosteroid signal regulated beta-tubulin GhTUB17 play a role in cotton fiber elongation by influencing microtubule organization

Cotton (Gossypium hirsutum L.) is an important natural fiber crop in the world. Cotton fiber cell is an extremely elongated single cell and a major cash product of cotton plants. Both brassinosteroids (BRs) and cytoskeleton play essential roles in fiber cell elongation. However, the relationship of BRs with cytoskeleton in fiber elongation is largely unknown. In the study, a BRs responsive beta-tubulin gene GhTUB17 was identified, which preferentially expressed in fiber cell and peaked in the rapid elongation stage. GhTUB17 was up- and down-regulated by brassinolide (BL) and brassinozole (BRZ), respectively. Furthermore, compared with control, GhTUB17 expression was increased in transgenic fiber cell overexpressing GhBES1, a transcription factor in BRs signaling, while was decreased in the fiber of li-1, a short fiber mutant of upland cotton. GhBES1 could bind to the promoter of GhTUB17 and positively regulate its expression. Overexpressing GhTUB17 promoted fiber elongation while suppressing GhTUB17 inhibited fiber elongation. The arrangement patterns of both microtubule and microfilament were altered in transgenic cotton fibers. These results indicated that GhTUB17 was a target gene of GhBES1 and play an important role in fiber cell elongation by regulating the organization of microtubule and microfilament cytoskeleton. It provided a link between BRs and cytoskeleton in fiber cell elongation and a potential gene to improve the fiber quality, and shed light on understanding the action mechanism of BRs.

Exogenous brassinolides and calcium chloride synergically maintain quality attributes of jujube fruit (Ziziphus jujuba Mill.)

Jujube fruit, rich in nutritional value, is commonly utilized in the processing of dried jujube fruit, jujube powder, concentrates, and fermented products, but improper storage may lead to quality cracking, diminishing the fruit's quality and commercial value. BR, known for its role as a plant growth regulator, is often applied to maintain postharvest fruit quality. The study investigated the impact of exogenous brassinolide (BR) and calcium chloride (CC) either alone or in combination on the quality attributes of jujube during storage. The results revealed that exogenous BC (BR combined with CC) effectively mitigated the senescence of jujube fruit. Specifically, the redden index, decay index, weight loss, and MDA content of jujube in the BC group decreased by 15 %, 8 %, 33 %, and 41 %, respectively, in comparison to the control group. Furthermore, all treatments maintained higher firmness, ascorbic acid content, total phenol content, chlorophyll content, and antioxidant capacity. Although exogenous BR was able to preserve the quality attributes of jujube, it exhibited instability. Treatment with CC alone was better than treatment with BR alone, but BC treatment maintained a better quality of jujube. The relative conductivity of jujube fruit in the BC group was consistently measured lower than that of the control group, with the exception of 8 d. Exogenous CC and BC were found to inhibit the decrease in total flavonoid and phenol content. Overall, the present study contributes to establishing a theoretical foundation for application of exogenous brassinolide combined with calcium chloride to maintain the postharvest quality attributes of fruit and vegetables.

SERK3A and SERK3B could be S-nitrosylated and enhance the salt resistance in tomato seedlings

Salinity hinders plant growth and development, resulting in reduced crop yields and diminished crop quality. Nitric oxide (NO) and brassinolides (BR) are plant growth regulators that coordinate a plethora of plant physiological responses. Nonetheless, the way in which these factors interact to affect salt tolerance is not well understood. BR is perceived by the BR receptor BRASSINOSTEROID INSENSITIVE 1 (BRI1) and its co-receptor BRI1-associated kinase 1 (BAK1) to form the receptor complex, eventually inducing BR-regulated responses. To response stress, a wide range of NO-mediated protein modifications is undergone in eukaryotic cells. Here, we showed that BR participated in NO-enhanced salt tolerance of tomato seedlings (Solanum lycopersicum cv. Micro-Tom) and NO may activate BR signaling under salt stress, which was related to NO-mediated S-nitrosylation. Further, in vitro and in vivo results suggested that BAK1 (SERK3A and SERK3B) was S-nitrosylated, which was inhibited under salt condition and enhanced by NO. Accordingly, knockdown of SERK3A and SERK3B reduced the S-nitrosylation of BAK1 and resulted in a compromised BR response, thereby abolishing NO-induced salt tolerance. Besides, we provided evidence for the interaction between BRI1 and SERK3A/SERK3B. Meanwhile, NO enhanced BRI1-SERK3A/SERK3B interaction. These results imply that NO-mediated S-nitrosylation of BAK1 enhances the interaction BRI1-BAK1, facilitating BR response and subsequently improving salt tolerance in tomato. Our findings illustrate a mechanism by which redox signaling and BR signaling coordinate plant growth in response to abiotic stress.

Tea green leafhopper infestations affect tea plant growth by altering the synthesis of brassinolide.

Tea green leafhoppers are insects widely distributed in major tea-growing areas. At present, less attention has been paid to the study on effect of tea green leafhopper infestation on tea growth phenotype. In this study, tea green leafhoppers were used to treat tea branches in laboratory and co-treated with brassinolide (BL), the highest bioactivity of brassinosteroids (BRs), in tea garden. The results showed that the expression of genes related to BRs synthesis was inhibited and BL content was reduced in tea shoots after infestation by tea green leafhoppers. In addition, area of each leaf position, length and diameter of internodes, and the biomass of the tender shoots of tea plant were decreased after infestation by tea green leafhoppers. The number of trichomes, leaf thickness, palisade tissue thickness and cuticle thickness of tea shoots were increased after tea green leafhoppers infestation. BL spraying could partially recover the phenotypic changes of tea branches caused by tea green leafhoppers infestation. Further studies showed that tea green leafhoppers infestation may regulate the expression of CsDWF4 (a key gene for BL synthesis) through transcription factors CsFP1 and CsTCP1a, which finally affect the BL content. Moreover, BL was applied to inhibit the tea green leafhoppers infestation on tea shoots. In conclusion, our study revealed the effect of plant hormone BL-mediated tea green leafhoppers infestation on the growth phenotype of tea plants.

Brassinosteroids biosynthetic gene MdBR6OX2 regulates salt stress tolerance in both apple and Arabidopsis

Salt stress is a critical limiting factor for fruit yield and quality of apples. Brassinosteroids (BRs) play an important role in response to abiotic stresses. In the present study, application of 2,4- Epicastasterone on seedlings of Malus ‘M9T337’ and Malus domestica ‘Gala3’ alleviated the physiological effects, such as growth inhibition and leaf yellowing, induced by salt stress. Further analysis revealed that treatment with NaCl induced expression of genes involved in BR biosynthesis in ‘M9T337’ and ‘Gala3’. Among which, the expression of BR biosynthetic gene MdBR6OX2 showed a three-fold upregulation upon salt treatment, suggesting its potential role in response to salt stress in apple. MdBR6OX2, belonging to the CYP450 family, contains a signal peptide region and a P450 domain. Expression patterns analysis showed that the expression of MdBR6OX2 can be significantly induced by different abiotic stresses. Overexpressing MdBR6OX2 enhanced the tolerance of apple callis to salt stress, and the contents of endogenous BR-related compounds, such as Typhastero (TY), Castasterone (CS) and Brassinolide (BL) were significantly increased in transgenic calli compared with that of wild-type. Extopic expression of MdBR6OX2 enhanced tolerance to salt stress in Arabidopsis. Genes associated with salt stress were significantly up-regulated, and the contents of BR-related compounds were significantly elevated under salt stress. Our data revealed that BR-biosynthetic gene MdBR6OX2 positively regulates salt stress tolerance in both apple calli and Arabidopsis.

Molecular mechanisms underlying low temperature inhibition of grain filling in maize (Zea mays L.): coordination of growth and cold responses.

Low temperature (LT) greatly restricts grain filling in maize (Zea mays L.), but the relevant molecular mechanisms are not fully understood. To better understand the effect of LT on grain development, 17 hybrids were subjected to LT stress in field trials over 3 years, and two hybrids of them with contrasting LT responses were exposed to 30/20°C and 20/10°C for 7 days during grain filling in a greenhouse. At LT, thousand-kernel weight declined, especially in LT-sensitive hybrid FM985, while grain-filling rate was on average about 48% higher in LT-tolerant hybrid DK159 than FM985. LT reduced starch synthesis in kernel mainly by suppression of transcript levels and enzyme activities for sucrose synthase and hexokinase. Brassinolide (BR) was abundant in DK159 kernel, and genes involved in BR and cytokinin signals were inducible by stress. LT downregulated the genes in light-harvesting complex and photosystem I/II subunits, accompanied by reduced photosynthetic rate and Fv/Fm in ear leaf. The LT-tolerant hybrid could maintain a high soluble sugar content and fast interconversion between sucrose and hexose in the stem internode and cob, improving assimilate allocation to kernel at LT stress and paving the way for simultaneous growth and LT stress responses.

DNA Methylation Analysis of Growth Differences between Upright and Inverted Cuttings of Populus yunnanensis.

DNA methylation is an important mechanism for epigenetic modifications that have been shown to be associated with responses to plant development. Previous studies found that inverted Populus yunnanensis cuttings were still viable and could develop into complete plants. However, the growth status of inverted cuttings was weaker than that of upright cuttings, and the sprouting time of inverted cuttings was later than that of upright cuttings. There is currently no research on DNA methylation patterns in inverted cuttings of Populus yunnanensis. In this study, we detected genome-wide methylation patterns of stem tips of Populus yunnanensis at the early growth stage and the rapid growth stage by Oxford Nanopore Technologies (ONT) methylation sequencing. We found that the methylation levels of CpG, CHG, CHH, and 6mA were 41.34%, 33.79%, 17.27%, and 12.90%, respectively, in the genome of inverted poplar cuttings, while the methylation levels of the four methylation types were higher in the genome of upright poplar cuttings than in inverted cuttings, 41.90%, 34.57%, 18.09%, and 14.11%, suggesting important roles for DNA methylation in poplar cells. In all comparison groups, CpG-type methylation genes in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were annotated to pathways associated with carbon metabolism, ribosome biogenesis in eukaryotes, glycolysis/gluconeogenesis, pyruvate metabolism, and mRNA detection pathways, suggesting that different biological processes are activated in upright and inverted cuttings. The results show that methylation genes are commonly present in the poplar genome, but only a few of them are involved in the regulation of expression in the growth and development of inverted cuttings. From this, we screened the DET2 gene for significant differences in methylation levels in upright or inverted cuttings. The DET2 gene is a key gene in the Brassinolide (BRs) synthesis pathway, and BRs have an important influence on the growth and development process of plants. These results provide important clues for studying DNA methylation patterns in P. yunnanensis.

Scientific Study of Brassinosteroids on Production of Fruit Crops: A Review

Brassinosteroids (BRs) are considered as the sixth group of plant growth regulators with significant growth promoting activity in many horticulture and agriculture crops. This phytohormone was discovered from the pollen of Brassica napus L. on the basis of their ability to promote growth. Currently, more than 70 BRs have been isolated from different plant species, but only three viz., Brassinolide (BL), 24-Epibrassinolide (EBL) and 28-Homobrassinolide (HBL) are found to be the most biologically active forms and extensively used in various crops. They are eco-friendly, effective at very low concentration, cost effective and almost always have synergetic effect when interacts with other phytohormones. Owing to diverse physiological roles and cross talk with other hormones influence on plant growth and development, they are being collectively referred as ‘pleiotropic phytohormone’. Although, BRs were discovered towards the end of twentieth century, but the dynamic applications and outcomes were reported mainly during the inception of twenty first century, hence BRs were termed as ‘hormone of the twenty-first century’. The application of BRs at different growth stages have been reported to positively influences such as nutrients uptakes, growth, yield and quality attributes on many fruit crops which includes tropical, sub-tropical and temperate fruit crops at very low concentrations have great roles in terms of growth, yield, fruit quality and stress tolerance mechanisms, which results to lucrative adventure of farming community. This review article will provide an elucidated insight into the application of BR in different fruit crops.