Strigolactone GR24-mediated mitigation of phosphorus deficiency through mycorrhization in aerobic rice.

Strigolactones (SLs) are a new class of plant hormones that play a significant role in regulating various aspects of plant growth promotion, stress tolerance and influence the rhizospheric microbiome. GR24 is a synthetic SL analog used in scientific research to understand the effects of SL on plants and to act as a plant growth promoter. This study aimed to conduct hormonal seed priming at different concentrations of GR24 (0.1, 0.5, 1.0, 5.0 and 10.0 µM with and without arbuscular mycorrhizal fungi (AMF) inoculation in selected aerobic rice varieties (CR Dhan 201, CR Dhan 204, CR Dhan 205, and CR Dhan 207), Kasalath-IC459373 (P-tolerant check), and IR-36 (P-susceptible check) under phosphorus (P)-deficient conditions to understand the enhancement of growth and priming effects in mycorrhization. Our findings showed that seed priming with 5.0 µM SL GR24 enhanced the performance of mycorrhization in CR Dhan 205 (88.91 %), followed by CR Dhan 204 and 207, and AMF sporulation in CR Dhan 201 (31.98 spores / 10 gm soil) and CR Dhan 207 (30.29 spores / 10 g soil), as well as rice growth. The study showed that the highly responsive variety CR Dhan 207 followed by CR Dhan 204, 205, 201, and Kasalath IC459373 showed higher P uptake than the control, and AMF treated with 5.0 µM SL GR24 varieties CR Dhan 205 followed by CR Dhan 207 and 204 showed the best performance in plant growth, chlorophyll content, and soil functional properties, such as acid and alkaline phosphatase activity, soil microbial biomass carbon (MBC), dehydrogenase activity (DHA), and fluorescein diacetate activity (FDA). Overall, AMF intervention with SL GR24 significantly increased plant growth, soil enzyme activity, and uptake of P compared to the control. Under P-deficient conditions, seed priming with 5.0 µM strigolactone GR24 and AMF inoculum significantly increased selected aerobic rice growth, P uptake, and soil enzyme activities. Application of SLs formulations with AMF inoculum in selected aerobic rice varieties, CR Dhan 207, CR Dhan 204, and CR Dhan 205, will play an important role in mycorrhization, growth, and enhancement of P utilization under P- nutrient deficient conditions.

Identification and expression of strigolactone biosynthesis and signaling genes and the in vitro effects of strigolactones in olive (Olea europaea L.).

Strigolactones (SLs), synthesized in plant roots, play a dual role in modulating plant growth and development, and in inducing the germination of parasitic plant seeds and arbuscular mycorrhizal fungi in the rhizosphere. As phytohormones, SLs are crucial in regulating branching and shaping plant architecture. Despite the significant impact of branching strategies on the yield performance of fruit crops, limited research has been conducted on SLs in these crops. In our study, we identified the transcript sequences of SL biosynthesis and signaling genes in olive (Olea europaea L.) using rapid amplification of cDNA ends. We predicted the corresponding protein sequences, analyzed their characteristics, and conducted molecular docking with bioinformatics tools. Furthermore, we quantified the expression levels of these genes in various tissues using quantitative real-time PCR. Our findings demonstrate the predominant expression of SL biosynthesis and signaling genes (OeD27, OeMAX3, OeMAX4, OeMAX1, OeD14, and OeMAX2) in roots and lateral buds, highlighting their importance in branching. Treatment with rac-GR24, an SL analog, enhanced the germination frequency of olive seeds in vitro compared with untreated embryos. Conversely, inhibition of SL biosynthesis with TIS108 increased lateral bud formation in a hard-to-root cultivar, underscoring the role of SLs as phytohormones in olives. These results suggest that modifying SL biosynthesis and signaling pathways could offer novel approaches for olive breeding, with potential applicability to other fruit crops.

Perception of butenolides by Bacillus subtilis via the α/ß hydrolase RsbQ.

The regulation of behavioral and developmental decisions by small molecules is common to all domains of life. In plants, strigolactones and karrikins are butenolide growth regulators that influence several aspects of plant growth and development, as well as interactions with symbiotic fungi.1,2,3 DWARF14 (D14) and KARRIKIN INSENSITIVE2 (KAI2) are homologous enzyme-receptors that perceive strigolactones and karrikins, respectively, and that require hydrolase activity to effect signal transduction.4,5,6,7 RsbQ, a homolog of D14 and KAI2 from the gram-positive bacterium Bacillus subtilis, regulates growth responses to nutritional stress via the alternative transcription factor SigmaB (σB).8,9 However, the molecular function of RsbQ is unknown. Here, we show that RsbQ perceives butenolide compounds that are bioactive in plants. RsbQ is thermally destabilized by the synthetic strigolactone GR24 and its desmethyl butenolide equivalent dGR24. We show that, like D14 and KAI2, RsbQ is a functional butenolide hydrolase that undergoes covalent modification of the catalytic histidine residue. Exogenous application of both GR24 and dGR24 inhibited the endogenous signaling function of RsbQ in vivo, with dGR24 being 10-fold more potent. Application of dGR24 to B. subtilis phenocopied loss-of-function rsbQ mutations and led to a significant downregulation of σB-regulated transcripts. We also discovered that exogenous butenolides promoted the transition from planktonic to biofilm growth. Our results suggest that butenolides may serve as inter-kingdom signaling compounds between plants and bacteria to help shape rhizosphere communities.

Strigolactone deficiency induces jasmonate, sugar and flavonoid phytoalexin accumulation enhancing rice defense against the blast fungus Pyricularia oryzae.

Strigolactones (SLs) are carotenoid-derived phytohormones that regulate plant growth and development. While root-secreted SLs are well-known to facilitate plant symbiosis with beneficial microbes, the role of SLs in plant interactions with pathogenic microbes remains largely unexplored. Using genetic and biochemical approaches, we demonstrate a negative role of SLs in rice (Oryza sativa) defense against the blast fungus Pyricularia oryzae (syn. Magnaporthe oryzae). We found that SL biosynthesis and perception mutants, and wild-type (WT) plants after chemical inhibition of SLs, were less susceptible to P. oryzae. Strigolactone deficiency also resulted in a higher accumulation of jasmonates, soluble sugars and flavonoid phytoalexins in rice leaves. Likewise, in response to P. oryzae infection, SL signaling was downregulated, while jasmonate and sugar content increased markedly. The jar1 mutant unable to synthesize jasmonoyl-l-isoleucine, and the coi1-18 RNAi line perturbed in jasmonate signaling, both accumulated lower levels of sugars. However, when WT seedlings were sprayed with glucose or sucrose, jasmonate accumulation increased, suggesting a reciprocal positive interplay between jasmonates and sugars. Finally, we showed that functional jasmonate signaling is necessary for SL deficiency to induce rice defense against P. oryzae. We conclude that a reduction in rice SL content reduces P. oryzae susceptibility by activating jasmonate and sugar signaling pathways, and flavonoid phytoalexin accumulation.

A multifaceted assessment of strigolactone GR24 and its derivatives: from anticancer and antidiabetic activities to antioxidant capacity and beyond.

Background: Strigolactones are signaling molecules produced by plants, the main functions are the intracorporeal control of plant development and plant growth. GR24 strigolactone is one of the synthetic strigolactones and due to its universality and easy availability, it is a standard and model compound for research on the properties and role of strigolactones in human health. Purpose: In this research work, the impact of mainly GR24 strigolactone on the human body and the role of this strigol-type lactone in many processes that take place within the human body are reviewed. Study design: The article is a review of publications on the use of GR24 strigolactone in studies from 2010-2023. Publications were searched using PubMed, Elsevier, Frontiers, and Springer databases. The Google Scholar search engine was also used. For the review original research papers and reviews related to the presented topic were selected. Results: The promising properties of GR24 and other strigolactone analogs in anti-cancer therapy are presented. Tumor development is associated with increased angiogenesis. Strigolactones have been shown to inhibit angiogenesis, which may enhance the anticancer effect of these γ-lactones. Furthermore, it has been shown that strigolactones have anti-inflammatory and antioxidant properties. There are also a few reports which show that the strigolactone analog may have antimicrobial and antiviral activity against human pathogens. Conclusion: When all of this is considered, strigolactones are molecules whose versatile action is their undeniable advantage. The development of research on these phytohormones makes it possible to discover their new, unique properties and surprising biological activities in relation to many mammalian cells.

Exogenous strigolactones alleviate drought stress in wheat (Triticum aestivum L.) by promoting cell wall biogenesis to optimize root architecture.

Exogenous strigolactones (SLs, GR24) are widely used to alleviate drought stress in wheat. The physiological and biochemical mechanisms via which SLs help overcome drought stress in wheat shoots have been reported; however, the mechanisms in wheat roots are unclear. The present study explored the effects of the exogenous application of SLs on wheat roots' growth and molecular responses under drought stress using physiological analysis and RNA-seq. RNA-seq of roots showed that SLs mainly upregulated signal transduction genes (SIS8, CBL3, GLR2.8, LRK10L-2.4, CRK29, and CRK8) and transcription factors genes (ABR1, BHLH61, and MYB93). Besides, SLs upregulated a few downstream target genes, including antioxidant genes (PER2, GSTF1, and GSTU6), cell wall biogenesis genes (SUS4, ADF3, UGT13248, UGT85A24, UGT709G2, BGLU31, and LAC5), an aquaporin-encoding gene (TIP4-3), and dehydrin-encoding genes (DHN2, DHN3, and DHN4). As a result, SLs reduced oxidative damage, optimized root architecture, improved leaf-water relation, and alleviated drought damage. Thus, the present study provides novel insights into GR24-mediated drought stress management and a scientific basis for proposing GR24 application.

Alleviation effect of GR24, a strigolactone analogue, on low-nitrogen stress in Malus baccata seedlings.

To investigate the efficacy of foliar application of GR24, a strigolactone analogue, in alleviating low-nitrogen stress in Malus baccata, we applied GR24 with different concentrations (0, 1, 5, 10, and 20 µmol·L-1) to leaves of plants under low nitrogen stress. We evaluated the changes in photosynthetic characteristics of leaves, reactive oxygen metabolism, and nitrogen assimilation in roots. The results showed that shoot biomass of seedling significantly decreased and root-shoot ratio increased under low-nitrogen stress. The chlorophyll contents decreased, the carotenoid content increased, and the photosynthetic activity decreased. The activities of superoxide dismutase and catalase enzymes in roots changed little, while the activities of peroxidase and ascorbic acid peroxidase enzymes, along with the levels of soluble sugar, free proline, and reactive oxygen species showed a significant increase, and the soluble protein content decreased. The NO3- content in roots decreased, the NH4+ content increased, while activities of nitrate reductase and glutamine synthase decreased. Compared to the control group without GR24 application, foliar sprays of 10 and 20 µmol·L-1 GR24 under both normal and low-nitrogen increased biomass and root-shoot ratio to varying degrees. Additionally, GR24 application increased chlorophyll content, photosynthesis indices (net photosynthetic rate, transpiration rate and stomatal conductance), and fluorescence (maximum photochemical efficiency of PSⅡ and quantum yield of electron transfer per unit area) performance parameters, as well as the contents of osmotic regulation substances (soluble protein, soluble sugar, and free proline) and glutamine synthase activity. Application of 10 and 20 µmol·L-1 GR24 under low-nitrogen stress decreased carotenoid, reactive oxygen species, and NH4+ contents, while increased the activities of antioxidases and key enzymes in nitrogen metabolism (nitrate reductase and glutamine synthase) and NO3- content. The 10 µmol·L-1 GR24 treatment was the most effective in alleviating low nitrogen stress, which has potential for application in apple orchards with low nitrogen soil.

Synthesis and Biological Profiling of Novel Strigolactone Derivatives for Arabidopsis Growth and Development.

The artificially synthesized strigolactone (SL) analogue GR24 is currently the most widely used reference compound in studying the biological functions of SLs. To elucidate the structure-activity relationship and find more promising derivatives with unique molecular profiles, we design and synthesized three series of novel GR24 derivatives and explored their activities in hypocotyl and root development of Arabidopsis. Among the 50 synthesized compounds, A11a, A12a, and A20d were found to have high activities comparable to GR24 for hypocotyl and/or primary root elongation inhibition in Arabidopsis. Some new analogues have been discovered to exhibit unique activities: (1) A20c, A21e, and A21o are specific inhibitors in primary root elongation; (2) A21c, A26c, and A27a exhibit a high promotion effect on Arabidopsis primary root elongation; and (3) A27e possesses the most unique profiles completely opposite to GR24 that promotes both hypocotyl elongation and primary root development. Moreover, we revealed that the AtD14 receptor does not affect the inhibitory effect of SL analogues in Arabidopsis root development. The ligand-receptor interactions for the most representative analogues A11a and A27e were deciphered with a long time scale molecular dynamics simulation study, which provides the molecular basis of their distinct functions, and may help scientists design novel phytohormones.

Strigolactone decreases cadmium concentrations by regulating cadmium localization and glyoxalase defense system: Effects on nodules organic acids and soybean yield.

To decrease environmental and human health risks associated with crop and soil contamination, alternative solutions are still needed. The information on strigolactones (SLs)-mediated elicitation of abiotic stress signaling and triggering physiological alterations is scarce in the plant. To unravel the same, soybean plants were subjected to cadmium (Cd) stress (20 mg kg-1), presence or absence of foliar applied SL (GR24) at the concentration of 10 µM. Excess Cd accumulation causes reduced growth (-52% shoot and +24% root), yield (-35%), physio-biochemical markers, organic acid production, and genes encoding heavy metal resilience in soybean. SL exogenous application decreased the growth and yield suppression (-12%), shielded chlorophyll (+3%), and prominently declined Cd-induced oxidative stress biomarkers accumulation in soybean. Moreover, SL effectively alleviates Cd-induced suppression in organic acids, superoxide dismutase (+73%), catalase activities (+117%), and increments ascorbate glutathione (ASA-GSH) cycle activities comprising ascorbate peroxidase, glutathione peroxidase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate reductase. SL-mediated upregulation of genes encoding heavy metals tolerance and glyoxalase defense system in Cd stressed plants. The results of this work point out that SL could be a promising player in mitigating Cd-induced injuries effectively in soybean. It acts through the antioxidant system modulation for redox homeostasis, shielding chloroplasts, enhancing photosynthetic apparatus, and elevating organic acid production in soybean plants.

Strigolactone Alleviates the Adverse Effects of Salt Stress on Seed Germination in Cucumber by Enhancing Antioxidant Capacity.

Strigolactones (SLs), as a new phytohormone, regulate various physiological and biochemical processes, and a number of stress responses, in plants. In this study, cucumber 'Xinchun NO. 4' is used to study the roles of SLs in seed germination under salt stress. The results show that the seed germination significantly decreases with the increase in the NaCl concentrations (0, 1, 10, 50, and 100 mM), and 50 mM NaCl as a moderate stress is used for further analysis. The different concentrations of SLs synthetic analogs GR24 (1, 5, 10, and 20 µM) significantly promote cucumber seed germination under NaCl stress, with a maximal biological response at 10 µM. An inhibitor of strigolactone (SL) synthesis TIS108 suppresses the positive roles of GR24 in cucumber seed germination under salt stress, suggesting that SL can alleviate the inhibition of seed germination caused by salt stress. To explore the regulatory mechanism of SL-alleviated salt stress, some contents, activities, and genes related to the antioxidant system are measured. The malondialdehyde (MDA), H2O2, O2-, and proline contents are increased, and the levels of ascorbic acid (AsA) and glutathione (GSH) are decreased under salt stress conditions, while GR24 treatment reduces MDA, H2O2, O2-, and proline contents, and increases AsA and GSH contents during seed germination under salt stress. Meanwhile, GR24 treatment enhances the decrease in the activities of antioxidant enzymes caused by salt stress [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX)], following which antioxidant-related genes SOD, POD, CAT, APX, and GRX2 are up-regulated by GR24 under salt stress. However, TIS108 reversed the positive effects of GR24 on cucumber seed germination under salt stress. Together, the results of this study revealed that GR24 regulates the expression levels of genes related to antioxidants and, therefore, regulates enzymatic activity and non-enzymatic substances and enhances antioxidant capacity, alleviating salt toxicity during seed germination in cucumber.

Metabolomic Analysis of Key Metabolites and Their Pathways Revealed the Response of Alfalfa (Medicago sativa L.) Root Exudates to rac-GR24 under Drought Stress.

The rac-GR24, an artificial analog of strigolactone, is known for its roles in inhibiting branches, and previous studies have reported that it has a certain mechanism to relieve abiotic stress, but the underlying metabolic mechanisms of mitigation for drought-induced remain unclear. Therefore, the objectives of this study were to identify associated metabolic pathways that are regulated by rac-GR24 in alfalfa (Medicago sativa L.) and to determine the metabolic mechanisms of rac-GR24 that are involved in drought-induced root exudate. The alfalfa seedling WL-712 was treated with 5% PEG to simulate drought, and rac-GR24 at a concentration of 0.1 µM was sprayed. After three days of treatment, root secretions within 24 h were collected. Osmotic adjustment substances and antioxidant enzyme activities were measured as physiological indicators, while LS/MS was performed to identify metabolites regulated by rac-GR24 of root exudate under drought. The results demonstrated that rac-GR24 treatment could alleviate the negative effects from drought-induced on alfalfa root, as manifested by increased osmotic adjustment substance content, cell membrane stability, and antioxidant enzyme activities. Among the 14 differential metabolites, five metabolites were uniquely downregulated in plants in rac-GR24 treatment. In addition, rac-GR24 could relieve drought-induced adverse effects on alfalfa through metabolic reprogramming in the pathways of the TCA cycle, pentose phosphate, tyrosine metabolism, and the purine pathway. This study indicated that rac-GR24 could improve the drought resistance of alfalfa by influencing the components of root exudates.

Zaxinone synthase controls arbuscular mycorrhizal colonization level in rice.

The Oryza sativa (rice) carotenoid cleavage dioxygenase OsZAS was described to produce zaxinone, a plant growth-promoting apocarotenoid. A zas mutant line showed reduced arbuscular mycorrhizal (AM) colonization, but the mechanisms underlying this behavior are unknown. Here, we investigated how OsZAS and exogenous zaxinone treatment regulate mycorrhization. Micromolar exogenous supply of zaxinone rescued root growth but not the mycorrhizal defects of the zas mutant, and even reduced mycorrhization in wild-type and zas genotypes. The zas line did not display the increase in the level of strigolactones (SLs) that was observed in wild-type plants at 7 days post-inoculation with AM fungus. Moreover, exogenous treatment with the synthetic SL analog GR24 rescued the zas mutant mycorrhizal phenotype, indicating that the lower AM colonization rate of zas is caused by a deficiency in SLs at the early stages of the interaction, and indicating that during this phase OsZAS activity is required to induce SL production, possibly mediated by the Dwarf14-Like (D14L) signaling pathway. OsZAS is expressed in arbuscule-containing cells, and OsPT11prom::OsZAS transgenic lines, where OsZAS expression is driven by the OsPT11 promoter active in arbusculated cells, exhibit increased mycorrhization compared with the wild type. Overall, our results show that the genetic manipulation of OsZAS activity in planta leads to a different effect on AM symbiosis from that of exogenous zaxinone treatment, and demonstrate that OsZAS influences the extent of AM colonization, acting as a component of a regulatory network that involves SLs.

Transcriptome analysis reveals the effects of strigolactone on shoot regeneration of apple.

KEY MESSAGE: We have demonstrated that strigolactone inhibitor, Tis108, could be used to improve shoot regeneration of apple, and provided insights into the molecular mechanism of strigolactone-mediated inhibition of adventitious shoot formation. Lack of an efficient transformation system largely stagnated the application of transgenic and CRISPR technology in apple rootstock. High shoot regeneration ability is an important basis for establishing an effective transformation system. In this study, we first demonstrated the inhibitory effects of strigolactones on the adventitious shoot formation of apple rootstock M26. Next, we successfully verified that strigolactone-biosynthesis inhibitor, Tis108, could be used to improve the shoot regeneration of woody plants. Our results also suggest strigolactone-biosynthesis gene, MdCCD7, can be a target gene for biotechnological improvements of shoot regeneration capacity. Furthermore, we have employed transcriptome analysis to reveal the molecular mechanism of strigolactone-mediated inhibition of adventitious shoot formation. Differentially expressed genes associated with photosynthesis, secondary growth, and organ development were identified. WGCNA suggests SLs might affect shoot regeneration through interaction with other hormones, especially, auxin, cytokinin, and ethylene. We were able to identify important candidate genes mediating the cross-talk between strigolactone and other hormones during the process of adventitious shoot formation. Overall, our findings not only propose a useful chemical for improving shoot regeneration in practice but also provide insights into the molecular mechanism of strigolactone-mediated inhibition of adventitious shoot formation.

Ancestral sequence reconstruction of the CYP711 family reveals functional divergence in strigolactone biosynthetic enzymes associated with gene duplication events in monocot grasses.

The strigolactone (SL) class of phytohormones shows broad chemical diversity, the functional importance of which remains to be fully elucidated, along with the enzymes responsible for the diversification of the SL structure. Here we explore the functional evolution of the highly conserved CYP711A P450 family, members of which catalyze several key monooxygenation reactions in the strigolactone pathway. Ancestral sequence reconstruction was utilized to infer ancestral CYP711A sequences based on a comprehensive set of extant CYP711 sequences. Eleven ancestral enzymes, corresponding to key points in the CYP711A phylogenetic tree, were resurrected and their activity was characterized towards the native substrate carlactone and the pure enantiomers of the synthetic strigolactone analogue, GR24. The ancestral and extant CYP711As tested accepted GR24 as a substrate and catalyzed several diversifying oxidation reactions on the structure. Evidence was obtained for functional divergence in the CYP711A family. The monocot group 3 ancestor, arising from gene duplication events within monocot grasses, showed both increased catalytic activity towards GR24 and high stereoselectivity towards the GR24 isomer resembling strigol-type SLs. These results are consistent with a role for CYP711As in strigolactone diversification in early land plants, which may have extended to the diversification of strigol-type SLs.

Strigolactones Modulate Salicylic Acid-Mediated Disease Resistance in Arabidopsis thaliana.

Strigolactones are low-molecular-weight phytohormones that play several roles in plants, such as regulation of shoot branching and interactions with arbuscular mycorrhizal fungi and parasitic weeds. Recently, strigolactones have been shown to be involved in plant responses to abiotic and biotic stress conditions. Herein, we analyzed the effects of strigolactones on systemic acquired resistance induced through salicylic acid-mediated signaling. We observed that the systemic acquired resistance inducer enhanced disease resistance in strigolactone-signaling and biosynthesis-deficient mutants. However, the amount of endogenous salicylic acid and the expression levels of salicylic acid-responsive genes were lower in strigolactone signaling-deficient max2 mutants than in wildtype plants. In both the wildtype and strigolactone biosynthesis-deficient mutants, the strigolactone analog GR24 enhanced disease resistance, whereas treatment with a strigolactone biosynthesis inhibitor suppressed disease resistance in the wildtype. Before inoculation of wildtype plants with pathogenic bacteria, treatment with GR24 did not induce defense-related genes; however, salicylic acid-responsive defense genes were rapidly induced after pathogenic infection. These findings suggest that strigolactones have a priming effect on Arabidopsis thaliana by inducing salicylic acid-mediated disease resistance.

Supplementation with rac-GR24 Facilitates the Accumulation of Biomass and Astaxanthin in Two Successive Stages of Haematococcus pluvialis Cultivation.

The unicellular freshwater green alga Haematococcus pluvialis has attracted much research attention due to its biosynthetic ability for large amounts of astaxanthin, a blood-red ketocarotenoid that is used in cosmetics, nutraceuticals, and pharmaceuticals. Recently, numerous studies have investigated the functions of natural astaxanthin; however, the high cost of the production of astaxanthin from H. pluvialis cultures restricts its commercial viability. There is an urgent need to fulfill commercial demands by increasing astaxanthin accumulation from H. pluvialis cultures. In this study, we discovered that treatment of H. pluvialis cultures at the beginning of the macrozooid stage (day 0) with 1 µM rac-GR24, a synthetic analogue of strigolactones (a class of phytohormones), led to significant increases in biomass [up to a maximum dry cell weight (DCW) of 0.53 g/L] during the macrozooid stage and astaxanthin (from 0.63 to 5.32% of DCW) during the hematocyst stage. We elucidated that this enhancement of biomass accumulation during the macrozooid stage by rac-GR24 is due to its increasing CO2 utilization efficiency in photosynthesis and carbohydrate biosynthesis. We also found that rac-GR24 stimulated the overproduction of nicotinamide adenine dinucleotide phosphate (NADPH) and antioxidant enzymes in H. pluvialis cultures, which alleviated the oxidative damage caused by reactive oxygen species generated during the hematocyst stage due to the exhaustion of nitrogen supplies. Moreover, rac-GR24 treatment of H. pluvialis synergistically altered the activity of the pathways of fatty acid biosynthesis and astaxanthin esterification, which resulted in larger amounts of astaxanthin being generated by rac-GR24-treated cultures than by controls. In summary, we have developed a feasible and economic rac-GR24-assisted strategy that increases the amounts of biomass and astaxanthin generated by H. pluvialis cultures, and have provided novel insights into the mechanistic roles of rac-GR24 to achieve these effects.

Transcriptional Analysis in the Arabidopsis Roots Reveals New Regulators that Link rac-GR24 Treatment with Changes in Flavonol Accumulation, Root Hair Elongation and Lateral Root Density.

The synthetic strigolactone (SL) analog, rac-GR24, has been instrumental in studying the role of SLs as well as karrikins because it activates the receptors DWARF14 (D14) and KARRIKIN INSENSITIVE 2 (KAI2) of their signaling pathways, respectively. Treatment with rac-GR24 modifies the root architecture at different levels, such as decreasing the lateral root density (LRD), while promoting root hair elongation or flavonol accumulation. Previously, we have shown that the flavonol biosynthesis is transcriptionally activated in the root by rac-GR24 treatment, but, thus far, the molecular players involved in that response have remained unknown. To get an in-depth insight into the changes that occur after the compound is perceived by the roots, we compared the root transcriptomes of the wild type and the more axillary growth2 (max2) mutant, affected in both SL and karrikin signaling pathways, with and without rac-GR24 treatment. Quantitative reverse transcription (qRT)-PCR, reporter line analysis and mutant phenotyping indicated that the flavonol response and the root hair elongation are controlled by the ELONGATED HYPOCOTYL 5 (HY5) and MYB12 transcription factors, but HY5, in contrast to MYB12, affects the LRD as well. Furthermore, we identified the transcription factors TARGET OF MONOPTEROS 5 (TMO5) and TMO5 LIKE1 as negative and the Mediator complex as positive regulators of the rac-GR24 effect on LRD. Altogether, hereby, we get closer toward understanding the molecular mechanisms that underlay the rac-GR24 responses in the root.

Involvement of strigolactone hormone in root development, influence and interaction with mycorrhizal fungi in plant: Mini-review.

Arbuscular mycorrhizal fungi (AMF) and plant symbiosis is the old, fascinating and beneficial relation that exist on earth for the plants. In this review, we have elaborated that the strigolactones (SLs) are released from the roots and function with root parasite, seeds and symbiotic AMF as contact chemicals. They are transported through the xylem in the plants and can regulate plant architecture, seed germination, nodule formation, increase the primary root length, influence the root hairs and physiological reactions to non-living agents by regulating their metabolism. SLs first evolved in ancient plant lineages as regulators of the basic production processes and then took a new role to maintain the growing biological complexities of terrestrial plant. SLs belongs to a diversified category of butenolide-bearing plant hormones related to various processes of agricultural concern. SLs also arouses the development of spores, the divergence and enlargement of hyphae of AMF, metabolism of mitochondria, reprogramming of transcription process, and generation of chitin oligosaccharides which further stimulate the early response of symbiosis in the host plant, results from better communication in plant and ability of coexistence with these fungi. The required nutrients are transferred from the roots to the shoots, which affect the physiological, biochemical, and morphological characteristics of the plant. On the other hand, the plant provides organic carbon in the form of sugars and lipids to the fungi, which they use as a source of energy and for carried out different anabolic pathways. SLs also lead to alteration in the dynamic and structure of actin in the root region as well as changes the auxin's transporter localization in the plasma membrane. Thus, this study reveals the functions that SLs play in the growth of roots, as well as their effect and interaction with AMF that promote plant growth.

Synthesis and Evaluation of New Halogenated GR24 Analogs as Germination Promotors for Orobanche cumana.

Orobanche and Striga are parasitic weeds extremely well adapted to the life cycle of their host plants. They cannot be eliminated by conventional weed control methods. Suicidal germination induced by strigolactones (SLs) analogs is an option to control these weeds. Here, we reported two new halogenated (+)-GR24 analogs, named 7-bromo-GR24 (7BrGR24) and 7-fluoro-GR24 (7FGR24), which were synthesized using commercially available materials following simple steps. Both compounds strongly promoted seed germination of Orobanche cumana. Their EC50 values of 2.3±0.28×10-8M (7BrGR24) and 0.97±0.29×10-8M (7FGR24) were 3- and 5-fold lower, respectively, than those of (+)-GR24 and rac-GR24 (EC50=5.1±1.32-5.3±1.44×10-8; p<0.05). The 7FGR24 was the strongest seed germination promoter tested, with a stimulation percentage of 62.0±9.1% at 1.0×10-8M and 90.9±3.8% at 1.0×10-6M. It showed higher binding affinity (IC50=0.189±0.012µM) for the SL receptor ShHTL7 than (+)-GR24 (IC50=0.248±0.032µM), rac-GR24 (IC50=0.319±0.032µM), and 7BrGR24 (IC50=0.521±0.087µM). Molecular docking experiments indicated that the binding affinity of both halogenated analogs to the strigolactone receptor OsD14 was similar to that of (+)-GR24. Our results indicate that 7FGR24 is a promising agent for the control of parasitic weeds.

GR24, A Synthetic Strigolactone Analog, and Light Affect the Organization of Cortical Microtubules in Arabidopsis Hypocotyl Cells.

Strigolactones are plant hormones regulating cytoskeleton-mediated developmental events in roots, such as lateral root formation and elongation of root hairs and hypocotyls. The latter process was addressed herein by the exogenous application of a synthetic strigolactone, GR24, and an inhibitor of strigolactone biosynthesis, TIS108, on hypocotyls of wild-type Arabidopsis and a strigolactone signaling mutant max2-1 (more axillary growth 2-1). Owing to the interdependence between light and strigolactone signaling, the present work was extended to seedlings grown under a standard light/dark regime, or under continuous darkness. Given the essential role of the cortical microtubules in cell elongation, their organization and dynamics were characterized under the conditions of altered strigolactone signaling using fluorescence microscopy methods with different spatiotemporal capacities, such as confocal laser scanning microscopy (CLSM) and structured illumination microscopy (SIM). It was found that GR24-dependent inhibition of hypocotyl elongation correlated with changes in cortical microtubule organization and dynamics, observed in living wild-type and max2-1 seedlings stably expressing genetically encoded fluorescent molecular markers for microtubules. Quantitative assessment of microscopic datasets revealed that chemical and/or genetic manipulation of strigolactone signaling affected microtubule remodeling, especially under light conditions. The application of GR24 in dark conditions partially alleviated cytoskeletal rearrangement, suggesting a new mechanistic connection between cytoskeletal behavior and the light-dependence of strigolactone signaling.