Gibberellins regulate masculinization through the SpGAI-SpSTM module in dioecious spinach.

The sex of dioecious plants is mainly determined by genetic factors, but it can also be converted by environmental cues such as exogenous phytohormones. Gibberellic acids (GAs) are well-known inducers of flowering and sexual development, yet the pathway of gibberellin-induced sex conversion in dioecious spinach (Spinacia oleracea L.) remains elusive. Based on sex detection before and after GA3 application using T11A and SSR19 molecular markers, we confirmed and elevated the masculinization effect of GA on a single female plant through exogenous applications of GA3, showing complete conversion and functional stamens. Silencing of GIBBERELLIC ACID INSENSITIVE (SpGAI), a single DELLA family protein that is a central GA signaling repressor, results in similar masculinization. We also show that SpGAI can physically interact with the spinach KNOX transcription factor SHOOT MERISTEMLESS (SpSTM), which is a homolog of the flower meristem identity regulator STM in Arabidopsis. The silencing of SpSTM also masculinized female flowers in spinach. Furthermore, SpSTM could directly bind the intron of SpPI to repress SpPI expression in developing female flowers. Overall, our results suggest that GA induces a female masculinization process through the SpGAI-SpSTM-SpPI regulatory module in spinach. These insights may help to clarify the molecular mechanism underlying the sex conversion system in dioecious plants while also elucidating the physiological basis for the generation of unisexual flowers so as to establish dioecy in plants.

GATA transcription factor in common bean: A comprehensive genome-wide functional characterization, identification, and abiotic stress response evaluation.

The GATA transcription factors (TFs) have been extensively studied for its regulatory role in various biological processes in many plant species. The functional and molecular mechanism of GATA TFs in regulating tolerance to abiotic stress has not yet been studied in the common bean. This study analyzed the functional identity of the GATA gene family in the P. vulgaris genome under different abiotic and phytohormonal stress. The GATA gene family was systematically investigated in the P. vulgaris genome, and 31 PvGATA TFs were identified. The study found that 18 out of 31 PvGATA genes had undergone duplication events, emphasizing the role of gene duplication in GATA gene expansion. All the PvGATA genes were classified into four significant subfamilies, with 8, 3, 6, and 13 members in each subfamily (subfamilies I, II, III, and IV), respectively. All PvGATA protein sequences contained a single GATA domain, but subfamily II members had additional domains such as CCT and tify. A total of 799 promoter cis-regulatory elements (CREs) were predicted in the PvGATAs. Additionally, we used qRT-PCR to investigate the expression profiles of five PvGATA genes in the common bean roots under abiotic conditions. The results suggest that PvGATA01/10/25/28 may play crucial roles in regulating plant resistance against salt and drought stress and may be involved in phytohormone-mediated stress signaling pathways. PvGATA28 was selected for overexpression and cloned into N. benthamiana using Agrobacterium-mediated transformation. Transgenic lines were subjected to abiotic stress, and results showed a significant tolerance of transgenic lines to stress conditions compared to wild-type counterparts. The seed germination assay suggested an extended dormancy of transgenic lines compared to wild-type lines. This study provides a comprehensive analysis of the PvGATA gene family, which can serve as a foundation for future research on the function of GATA TFs in abiotic stress tolerance in common bean plants.

Genetic analysis of late-maturity α-amylase in twelve wheat populations.

MAIN CONCLUSION: Genetic loci, particularly those with an effect in the independent panel, could be utilised to further reduce LMA expression when used with favourable combinations of genes known to affect LMA. Late maturity α-amylase (LMA) is a grain quality defect involving elevated α-amylase within the aleurone of wheat (Triticum aestivum L.) grains. The genes known to affect expression are the reduced height genes Rht-B1 (chromosome 4B) and Rht-D1 (chromosome 4D), and an ent-copalyl diphosphate synthase gene (LMA-1) on chromosome 7B. Other minor effect loci have been reported, but these are poorly characterised and further genetic understanding is needed. In this study, twelve F4-derived populations were created through single seed descent, genotyped and evaluated for LMA. LMA-1 haplotype C and the Rht-D1b allele substantially reduced LMA expression. The alternative dwarfing genes Rht13 and Rht18 had no significant effect on LMA expression. Additional quantitative trait loci (QTL) were mapped at 16 positions in the wheat genome. Effects on LMA expression were detected for four of these QTL in a large independent panel of Australian wheat lines. The QTL detected in mapping populations and confirmed in the large independent panel provide further opportunity for selection against LMA, especially if combined with Rht-D1b and/or favourable haplotypes of LMA-1.

A natural variation in OsDSK2a modulates plant growth and salt tolerance through phosphorylation by SnRK1A in rice.

Plants grow rapidly for maximal production under optimal conditions; however, they adopt a slower growth strategy to maintain survival when facing environmental stresses. As salt stress restricts crop architecture and grain yield, identifying genetic variations associated with growth and yield responses to salinity is critical for breeding optimal crop varieties. OsDSK2a is a pivotal modulator of plant growth and salt tolerance via the modulation of gibberellic acid (GA) metabolism; however, its regulation remains unclear. Here, we showed that OsDSK2a can be phosphorylated at the second amino acid (S2) to maintain its stability. The gene-edited mutant osdsk2aS2G showed decreased plant height and enhanced salt tolerance. SnRK1A modulated OsDSK2a-S2 phosphorylation and played a substantial role in GA metabolism. Genetic analysis indicated that SnRK1A functions upstream of OsDSK2a and affects plant growth and salt tolerance. Moreover, SnRK1A activity was suppressed under salt stress, resulting in decreased phosphorylation and abundance of OsDSK2a. Thus, SnRK1A preserves the stability of OsDSK2a to maintain plant growth under normal conditions, and reduces the abundance of OsDSK2a to limit growth under salt stress. Haplotype analysis using 3 K-RG data identified a natural variation in OsDSK2a-S2. The allele of OsDSK2a-G downregulates plant height and improves salt-inhibited grain yield. Thus, our findings revealed a new mechanism for OsDSK2a stability and provided a valuable target for crop breeding to overcome yield limitations under salinity stress.

GhSBI1, a CUP-SHAPED COTYLEDON 2 homologue, modulates branch internode elongation in cotton.

Branch length is an important plant architecture trait in cotton (Gossypium) breeding. Development of cultivars with short branch has been proposed as a main object to enhance cotton yield potential, because they are suitable for high planting density. Here, we report the molecular cloning and characterization of a semi-dominant quantitative trait locus, Short Branch Internode 1(GhSBI1), which encodes a NAC transcription factor homologous to CUP-SHAPED COTYLEDON 2 (CUC2) and is regulated by microRNA ghr-miR164. We demonstrate that a point mutation found in sbi1 mutants perturbs ghr-miR164-directed regulation of GhSBI1, resulting in an increased expression level of GhSBI1. The sbi1 mutant was sensitive to exogenous gibberellic acid (GA) treatments. Overexpression of GhSBI1 inhibited branch internode elongation and led to the decreased levels of bioactive GAs. In addition, gene knockout analysis showed that GhSBI1 is required for the maintenance of the boundaries of multiple tissues in cotton. Transcriptome analysis revealed that overexpression of GhSBI1 affects the expression of plant hormone signalling-, axillary meristems initiation-, and abiotic stress response-related genes. GhSBI1 interacted with GAIs, the DELLA repressors of GA signalling. GhSBI1 represses expression of GA signalling- and cell elongation-related genes by directly targeting their promoters. Our work thus provides new insights into the molecular mechanisms for branch length and paves the way for the development of elite cultivars with suitable plant architecture in cotton.

C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 contributes to GA-mediated growth and flowering by interaction with DELLA proteins.

Gibberellic acid (GA) plays a central role in many plant developmental processes and is crucial for crop improvement. DELLA proteins, the core suppressors in the GA signaling pathway, are degraded by GA via the 26S proteasomal pathway to release the GA response. However, little is known about the phosphorylation-mediated regulation of DELLA proteins. In this study, we combined GA response assays with protein-protein interaction analysis to infer the connection between Arabidopsis thaliana DELLAs and the C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 (CPL3), a phosphatase involved in the dephosphorylation of RNA polymerase II. We show that CPL3 directly interacts with DELLA proteins and promotes DELLA protein stability by inhibiting its degradation by the 26S proteasome. Consequently, CPL3 negatively modulates multiple GA-mediated processes of plant development, including hypocotyl elongation, flowering time, and anthocyanin accumulation. Taken together, our findings demonstrate that CPL3 serves as a novel regulator that could improve DELLA stability and thereby participate in GA signaling transduction.

Meiotic instability and irregular chromosome pairing underpin heat-induced infertility in bread wheat carrying the Rht-B1b or Rht-D1b Green Revolution genes.

Mutations in the Rht-B1a and Rht-D1a genes of wheat (Triticum aestivum; resulting in Rht-B1b and Rht-D1b alleles) cause gibberellin-insensitive dwarfism and are one of the most important elements of increased yield introduced during the 'Green Revolution'. We measured the effects of a short period of heat imposed during the early reproductive stage on near-isogenic lines carrying Rht-B1b or Rht-D1b alleles, with respect to the wild-type (WT). The temperature shift caused a significant fertility loss within the ears of Rht-B1b and Rht-D1b wheats, greater than that observed for the WT. Defects in chromosome synapsis, reduced homologous recombination and a high frequency of chromosome mis-segregation were associated with reduced fertility. The transcription of TaGA3ox gene involved in the final stage of gibberellic acid (GA) biosynthesis was activated and ultra-performance liquid chromatography-tandem mass spectrometry identified GA1 as the dominant bioactive GA in developing ears, but levels were unaffected by the elevated temperature. Rht-B1b and Rht-D1b mutants were inclined to meiotic errors under optimal temperatures and showed a higher susceptibility to heat than their tall counterparts. Identification and introduction of new dwarfing alleles into modern breeding programmes is invaluable in the development of climate-resilient wheat varieties.

GhGASA14 regulates the flowering time of upland cotton in response to GA3.

KEY MESSAGE: The silencing of GhGASA14 and the identification of superior allelic variation in its coding region indicate that GhGASA14 may positively regulate flowering and the response to GA3. Gibberellic acid-stimulated Arabidopsis (GASA), a member of the gibberellin-regulated short amino acid family, has been extensively investigated in several plant species and found to be critical for plant growth and development. However, research on this topic in cotton has been limited. In this study, we identified 38 GhGASAs that were dispersed across 18 chromosomes in upland cotton, and all of these genes had a GASA core domain. Transcriptome expression patterns and qRT-PCR results revealed that GhGASA9 and GhGASA14 exhibited upregulated expression not only in the floral organs but also in the leaves of early-maturing cultivars. The two genes were functionally characterized by virus-induced gene silencing (VIGS), and the budding and flowering times after silencing the target genes were later than those of the control (TRV:00). Compared with that in the water-treated group (MOCK), the flowering period of the different fruiting branches in the GA3-treated group was more concentrated. Interestingly, allelic variation was detected in the coding sequence of GhGASA14 between early-maturing and late-maturing accessions, and the frequency of this favorable allele was greater in high-latitude cotton cultivars than in low-latitude ones. Additionally, a significant linear relationship was observed between the expression level of GhGASA14 and flowering time among the 12 upland cotton accessions. Taken together, these results indicated that GhGASA14 may positively regulate flowering time and respond to GA3. These findings could lead to the use of valuable genetic resources for breeding early-maturing cotton cultivars in the future.

Gibberellic acid signaling promotes resistance to saline-alkaline stress by increasing the uptake of ammonium in rice.

Gibberellic acid (GA) plays important roles in diverse biological processes in plants. However, its function in rice (Oryza sativa) resistance to saline-alkaline (SAK) stress is unclear. This study showed that SAK stimuli changed GA signaling gene expression levels. Genetic analyses using the mutants of key GA signaling regulators, Slender rice 1 (SLR1) and Dwarf 1(D1), demonstrated that SLR1 negatively, while D1 positively regulated the resistance of rice to SAK stress, suggesting that the GA signaling positively regulates the resistance of rice to SAK. Further analyses revealed that SLR1 interacted with and inhibited the transcription activation activity of IDD10 and bZIP23. Furthermore, IDD10 interacted with bZIP23 to activate Ammonium transporter 1;2 (AMT1;2), and slr1, IDD10 OX and bZIP23 OX accumulated more ammonium (NH4+), while idd10 and bzip23 accumulated less NH4+ than the wild-type (WT). In addition, the bzip23 mutant was more sensitive to SAK, while bZIP23 OX was less sensitive compared with the WT, suggesting that bZIP23 positively regulates the resistance of rice to SAK. These findings demonstrate that GA signaling promoted rice's SAK resistance by regulating NH4+ uptake through the SLR1-IDD10-bZIP23 pathway.

Plant hormone profiling of scion and rootstock incision sites and intra- and inter-family graft junctions in Nicotiana benthamiana.

Many previous studies have suggested that various plant hormones play essential roles in the grafting process. In this study, to understand the plant hormones that accumulate in the graft junctions, whether these are supplied from the scion or rootstock, and how these hormones play a role in the grafting process, we performed a hormonome analysis that accumulated in the incision site of the upper plants from the incision as "ungrafted scion" and lower plants from the incision as "ungrafted rootstock" in Nicotiana benthamiana. The results revealed that indole-3-acetic acid (IAA) and gibberellic acid (GA), which regulate cell division; abscisic acid (ABA) and jasmonic acid (JA), which regulate xylem formation; cytokinin (CK), which regulates callus formation, show different accumulation patterns in the incision sites of the ungrafted scion and rootstock. In addition, to try discussing the differences in the degree and speed of each event during the grafting process between intra- and inter-family grafting by determining the concentration and accumulation timing of plant hormones in the graft junctions, we performed hormonome analysis of graft junctions of intra-family grafted plants with N. benthamiana as scion and Solanum lycopersicum as rootstock (Nb/Sl) and inter-family grafted plants with N. benthamiana as scion and Arabidopsis thaliana as rootstock (Nb/At), using the ability of Nicotiana species to graft with many plant species. The results revealed that ABA and CK showed different accumulation timings; IAA, JA, and salicylic acid (SA) showed similar accumulation timings, while different accumulated concentrations in the graft junctions of Nb/Sl and Nb/At. This information is important for understanding the molecular mechanisms of plant hormones in the grafting process and the differences in molecular mechanisms between intra- and inter-family grafting.

Genetic study for seed germination and shattering in Euphorbia lagascae in response to different seed treatments.

Euphorbia lagascae Spreng is a promising emerging oilseed crop, with its seed oil accounting for approximately 50% of the seed weight. Euphorbia oil contains a significant amount of vernolic acid, comprising two-thirds of its composition, which boasts various industrial applications, including acting as a stabilizer-plasticizer and natural dye. However, this species was known to have a high degree of seed-shattering and a low germination rate, which act as two important barriers to large-scale production and exploitation. Therefore, the objective of this study is to determine the genetic control of seed germination and seed-shattering traits in order to develop a reliable pipeline that would be applicable for industries and breeders to select superior E. lagascae lines and design a robust breeding scheme in a short time at reduced labor costs. For this objective, five different wild-type genotypes of E. lagascae that demonstrated high germination potential were crossed with an ethyl methanesulfonate (EMS) mutant genotype that produces non-shattering capsules. The F2 populations from two successful crosses (A and B) were separated into three different treated groups for seed germination evaluation and to study the segregation of 200 individuals per F2 population. The three treatments were: light, gibberellic acid (GA3), and control treatment. Consequently, plants treated with approximately 250 µmol/m2/s of light showed significant improvement in germination up to 75% in cross A and 82.4 % in cross B compared with the control plants and the group treated with 0.05% GA3. According to the chi-square test results, the inheritance pattern of seed germination in response to light treatment follows a 3:1 segregation ratio between germinated and non-germinated seeds, indicating a dominant gene action in the F2 generation. The same conclusion was followed for the shattering trait in the group treated with light, which was also simply inherited as a 3:1 ratio for shattering vs. non-shattering capsules. Our results emphasize the importance and significance of light treatment in producing uniform populations through acceptable germination and shattering resistance of the mutant genotypes of E. lagascae. This is the first report of light treatment that significantly improved seed germination of E. lagascae, which may enhance efforts in the development of this new industrial crop as a feedstock for vernolic acid production.

Brassinosteroid decreases cadmium accumulation via regulating gibberellic acid accumulation and Cd fixation capacity of root cell wall in rice (Oryza sativa).

The precise mechanism behind the association between plants' reactions to cadmium (Cd) stress and brassinosteroid (BR) remains unclear. In the current investigation, Cd stress quickly increased the endogenous BR concentration in the rice roots. Exogenous BR also increased the hemicellulose level in the root cell wall, which in turn increased its capacity to bind Cd. Simultaneously, the transcription level of genes responsible for root Cd absorption was decreased, including Natural Resistance-Associated Macrophage Protein 1/5 (OsNRAMP1/5) and a major facilitator superfamily gene called OsCd1. Ultimately, the increased expression of Heavy Metal ATPase 3 (OsHMA3) and the decreased expression of OsHMA2, which was in charge of separating Cd into vacuoles and translocating Cd to the shoots, respectively, led to a decrease in the amount of Cd that accumulated in the rice shoots. In contrast, transgenic rice lines overexpressing OsGSK2 (a negative regulator in BR signaling) accumulated more Cd, while OsGSK2 RNA interference (RNAi) rice line accumulated less Cd. Furthermore, BR increased endogenous Gibberellic acid (GA) level, and applying GA could replicate its alleviative effect. Taken together, BR decreased Cd accumulation in rice by mediating the cell wall's fixation capacity to Cd, which might relied on the buildup of the GA.

Biochemical properties and pigment contents of Satureja genotypes affected by plant growth regulators and temperature stress.

There is little data, to our knowledge, on the biochemical properties of different Satureja sp. genotypes affected by plant growth regulators (PGR) under temperature stress. A split plot research on the basis of a complete randomized block design with three replicates examining temperature stress (planting dates, 8th of April, May and June) (main factor), and the factorial combination of plant growth regulators (PGR, control (CO), gibberellic acid (GA), fertilization (MI), and amino acid (A)), and genotypes (Khuzestani, Mutika, and Bakhtiari) on plant biochemical properties, was conducted. Plant pigment contents (chlorophyll a, and b and carotenoids (car)), antioxidant activity (catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (GP)), and leaf protein were determined. Treatments significantly and differently affected the genotypes performance. PD3 and PD1resulted in significantly higher activity of APX (0.059 U. mg-1) and GP (0.190 U. mg-1), respectively (P ≤ 0.05). Temperature stress significantly affected plant CAT activity (U. mg-1) at PD1 (0.084) and PD3 (0.820). Higher temperature significantly enhanced leaf Pro, MI increased plant APX (0.054) and CAT activities (0.111 U. mg-1) significantly, and GA resulted in the highest and significantly different GP activity (0.186 U. mL-1). Treatments T1 and T3 significantly enhanced Chla and Car content, and MI resulted in significantly higher Chlb content (0.085 mg g-1 leaf fresh weight). Car and CAT are the two most sensitive biochemical traits under temperature stress and can more effectively regulate Satureja growth and activity. It is possible to alleviate temperature stress on Satureja biochemical properties by the tested PGR.

Drought-Tolerant Bacteria and Arbuscular Mycorrhizal Fungi Mitigate the Detrimental Effects of Drought Stress Induced by Withholding Irrigation at Critical Growth Stages of Soybean (Glycine max, L.).

Considering current global climate change, drought stress is regarded as a major problem negatively impacting the growth of soybeans, particularly at the critical stages R3 (early pod) and R5 (seed development). Microbial inoculation is regarded as an ecologically friendly and low-cost-effective strategy for helping soybean plants withstand drought stress. The present study aimed to isolate newly drought-tolerant bacteria from native soil and evaluated their potential for producing growth-promoting substances as well as understanding how these isolated bacteria along with arbuscular mycorrhizal fungi (AMF) could mitigate drought stress in soybean plants at critical growth stages in a field experiment. In this study, 30 Bradyrhizobium isolates and 30 rhizobacterial isolates were isolated from the soybean nodules and rhizosphere, respectively. Polyethylene glycol (PEG) 6000 was used for evaluating their tolerance to drought, and then the production of growth promotion substances was evaluated under both without/with PEG. The most effective isolates (DTB4 and DTR30) were identified genetically using 16S rRNA gene. A field experiment was conducted to study the impact of inoculation with DTB4 and DTR30 along with AMF (Glomus clarum, Funneliformis mosseae, and Gigaspora margarita) on the growth and yield of drought-stressed soybeans. Our results showed that the bioinoculant applications improved the growth traits (shoot length, root length, leaf area, and dry weight), chlorophyll content, nutrient content (N, P, and K), nodulation, and yield components (pods number, seeds weight, and grain yield) of soybean plants under drought stress (p ≤ 0.05). Moreover, proline contents were decreased due to the bioinoculant applications under drought when compared to uninoculated treatments. As well as the count of bacteria, mycorrhizal colonization indices, and the activity of soil enzymes (dehydrogenase and phosphatase) were enhanced in the soybean rhizosphere under drought stress. This study's findings imply that using a mixture of bioinoculants may help soybean plants withstand drought stress, particularly during critical growth stages, and that soybean growth, productivity, and soil microbial activity were improved under drought stress.

Liposome-encapsulated cytochrome P450 and gibberellic acid biosynthesis in Priestia megaterium RP1.

This study explores the potential of liposome encapsulated silica immobilized cytochrome P450 monooxygenase (LSICY) for bioremediation of mercury (Hg2+). Current limitations in Hg2+ reduction, including sensitivity to factors like pH and cost, necessitate alternative methods. We propose LSICY as a solution, leveraging the enzymatic activities of cytochrome P450 monooxygenase (CYPM) for Hg2+ reduction through hydroxylation and oxygenation. Our investigation employs LSICY to assess its efficacy in mitigating Hg2+ toxicity in Oryza sativa (rice) plants. Gas chromatography confirmed gibberellic acid (GA) presence in the Hg2+ reducing bacteria Priestia megaterium RP1 (PMRP1), highlighting a potential link between CYP450 activity and plant health. This study demonstrates the promise of LSICY as a sustainable and effective approach for Hg2+ bioremediation, promoting a safer soil environment.

Gibberellic acid targeting ZBTB16 reduces NF-κB dependent inflammatory stress in sepsis-induced neuroinflammation.

OBJECTIVE: Sepsis is frequently complicated by neuroinflammation. Gibberellic acid (GA3) is recognized for its anti-inflammatory properties. In this study, our objective was to investigate whether GA3 could alleviate Nuclear factor-kappa B (NF-κB) -dependent inflammatory stress in sepsis-induced neuroinflammation. METHODS: C57BL/6 J mice were administered 10 mg/kg lipopolysaccharide (LPS) to induce sepsis. BV2 cells were pre-incubated with GA3 and subjected lipopolysaccharide stimulation to replicate the inflammatory microglia during sepsis. Subsequently, we assessed the release of IL-6, TNF-α, and IL-1ß, along with the expression of Zbtb16, NF-κB, and IκB. To investigate whether any observed anti-inflammatory effects of GA3 were mediated through a Zbtb16-dependent mechanism, Zbtb16 was silenced using siRNA. RESULTS: GA3 improved the survival of sepsis mice and alleviated post-sepsis cognitive impairment. Additionally, GA3 attenuated microglial M1 activation (pro-inflammatory phenotype), inflammation, and neuronal damage in the brain. Moreover, GA3 inhibited the release of TNF-α, IL-6, and IL-1ß in microglia stimulated with LPS. The NF-κB signaling pathway emerged as one of the key molecular pathways associated with the impact of GA3 on LPS-stimulated microglia. Lastly, GA3 upregulated Zbtb16 expression in microglia that had been downregulated by LPS. The inhibitory effects of GA3 on microglial M1 activation were partially reversed through siRNA knockdown of Zbtb16. CONCLUSIONS: Pre-incubation of microglia with GA3 led to the upregulation of the NF-κB regulator, Zbtb16. This process counteracted LPS-induced microglial M1 activation, resulting in an anti-inflammatory effect upon subsequent LPS stimulation.

Functional Characterization and Localization of Plant-Growth Promoting Bacteria Grown Under Stressful Conditions.

Various bacterial species are associated with plant roots. However, symbiotic and free-living plant growth-promoting bacteria (PGPB) can only help plants to grow and develop under normal and stressful conditions. Several biochemical and in vitro assays were previously designed to differentiate between the PGPB and other plant-associated bacterial strains. This chapter describes and summarizes some of these assays and proposes a strategy to screen for PGPB. To determine the involvement of the PGPB in abiotic stress tolerance, assays for the ability to produce 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, ammonium, gibberellic acid (GA), indole acetic acid (IAA), and microbial volatile organic compounds (mVOCs) are described in this chapter. Additionally, assays to show the capacity to solubilize micronutrients such as potassium, phosphorus, and zinc by bacteria were also summarized in this chapter. To determine the contribution of the PGPB in biotic stress tolerance in plants, Fe-siderophore, hydrogen cyanide, and antibiotic and antifungal metabolites production assays were described. Moreover, assays to investigate the growth-promotion activities of a bacterium strain on plants, using the gnotobiotic root elongation, in vitro, and pots assays, were explained. Finally, an assay for the localization of endophytic bacterium in plant tissues was also presented in this chapter. Although the assays described in this chapter can give evidence of the nature of the mechanism behind the PGPB actions, other unknown growth-promoting means are yet to decipher, and until then, new methodologies will be developed.

Gibberellic acid induced oxidative stress, endoplasmic reticulum stress, and apoptosis in the livers of gibel carp (Carassius auratus gibelio).

Gibberellic acid (GA3), one of the most plant growth stimulator, is widely applied in agricultural regions and in beer industry. However, GA3 residue remained in soil and water can cause toxicity to all organisms. In this study, we investigated the mechanisms of GA3-induced hepatic injury in gibel carp (Carassius auratus gibelio). We found that GA3 exposure caused oxidative stress, endoplasmic reticulum stress (ERS), and apoptosis. The gibel carp exposed to GA3 exhibited significant alteration in erythrocyte nuclei. GA3 induced liver damage, as indicated by increasing the aminopherase activities. GA3 led to oxidative stress by increasing malondialdehyde content and decreasing the activities of CAT and GPx. GA3 stimulated ERS and increased the expression of grp78, perk, eif2s1α, chop, atf4, ire1α, xbp1, and atf6. Additionally, GA3 down-regulated the level of anti-apoptotic gene Bcl-2 and up-regulated the levels of pro-apoptotic genes bax and caspase-3. Overall results demonstrated that GA3 caused hepatic injury in gibel carp by increasing oxidative stress, ERS, and apoptosis.

Improving the productivity of gibberellic acid by combining small-molecule compounds-based targeting technology and transcriptomics analysis in Fusarium fujikuroi.

Gibberellic acid (GA3), produced industrially by Fusarium fujikuroi, stands as a crucial plant growth regulator extensively employed in the agriculture filed while limited understanding of the global metabolic network hinders researchers from conducting rapid targeted modifications. In this study, a small-molecule compounds-based targeting technology was developed to increase GA3 production. Firstly, various small molecules were used to target key nodes of different pathways and the result displayed that supplement of terbinafine improved significantly GA3 accumulation, which reached to 1.08 g/L. Subsequently, lipid and squalene biosynthesis pathway were identified as the key pathways influencing GA3 biosynthesis by transcriptomic analysis. Thus, the strategies including in vivo metabolic engineering modification and in vitro supplementation of lipid substrates were adopted, both contributed to an enhanced GA3 yield. Finally, the engineered strain demonstrated the ability to achieve a GA3 yield of 3.24 g/L in 5 L bioreactor when utilizing WCO as carbon source and feed.

Synergistic effect of selenium and gibberellic acid for enhanced biomass, lipid and improved biodiesel quality from Tetradesmus obliquus through response surface methodology.

Biodiesel production from microalgae presents an innovative solution for renewable energy. This study investigates biodiesel production using Tetradesmus obliquus ON506010.1 by optimizing substrates, selenium and gibberellic acid. Using 15 µg/L selenium, lipid content and biomass productivity reached 35.45 %±0.92 and 0.178 g/L/day ± 0.051. With 50 µM gibberellic acid, biomass productivity and lipid content peaked at 0.785 ± 0.101 g/L/day and 38.95 %±0.35, surpassing the control. Fatty acid composition, biodiesel properties, and mRNA expression of lipid synthesis enzymes (acetyl CoA carboxylase (ACC) and fatty acid desaturase (FAD)) correlated. Combining 10 µg/L selenium with 75 µM gibberellic acid with response surface methodology (RSM) increased lipid content (42.80 % ±0.11) and biomass productivity (0.964 g/L/day ± 0.128). ACC and FAD upregulation validated this enhancement, with a 4.4-fold increase in FAD expression. Fatty acid composition and most biodiesel properties met international standards demonstrating Tetradesmus obliquus ON506010.1's potential for sustainable biodiesel production with better cold flow property and oxidative stability.