Gibberellin Production Research Articles

Expression difference of P450–1 and P450–4 between G- and F-groups of Fusarium fujikuroi

Fusarium fujikuroi is the pathogen of rice bakanae disease and has been intensively studied for gibberellin (GA) production. F. fujikuroi is phylogenetically subclassified into G- and F-groups, which differ in GA and fumonisin production. A higher amount of GA is produced by the G-group than the F-group. A previous study found that the GA production difference between the G-group strain Gfc0801001 and the F-group strain Gfc0825009 was due to the GA gene cluster. In this study, higher expression of P450–1 and P450–4 in the GA gene cluster were detected in G-group strains than in F-group strains. The nucleotide sequence of the bidirectional promoter region between P450–1 and P450–4 and expression of P450–1 and P450–4 were compared between G- and F-group strains. The promoter sequence was identical among G-group strains while nucleotide substitutions were detected at various locations in F-group strains. Most F-group strains did not share the same substitutions. These results combined with the ancestral position of the F-group in the phylogenetic tree suggest that the population carrying highly expressed sequence generates the G-group rather than the low expression of P450–1 and P450–4 in the F-group due to a specific mutation occured in G-group.

Mastery of Sumber Urip-1 farmer group in Malang Regency in the production and application of organic auxins and gibberellins

Organic agriculture was a manifestation of sustainable agriculture as stipulated in SDG 2. Therefore, farmer assistance activities were needed to implement organic agricultural applications, namely the manufacture and use of organic auxins and gibberellins. This communityservice aimed to provide assistance in the manufacture and use of organic auxins and gibberellins. The method in implementing this activity was direct assistance and practice to solve problems. The activity was held for three months, in Wonorejo Village, Poncokusumo District, Malang Regency, East Java Province, with a partner named Sumber Urip-1 Farmer Group. The indicator for the success of community service activities was that> 75% of the members (out of a total of 20 people) can practice making organic auxins and gibberellins. To improve mentoring performance, there was team synergy in the form of division of labor, cooperation patterns, and collaboration patterns. Evaluation and monitoring of activities was carried out by holding morning meetings every Friday. Through a series of activities, partners have mastered the methods of (1) manufacturing and (2) using organic auxins and gibberellins. Products were branded with the brands "Vignauksin" (from bean sprouts), "Fulicauksin" (from snails), and "Zeaberelin" (from corn). Partners' mastery of the methods of making and using organic auxins and gibberellins will support organic farming efforts which were the embodiment of sustainable agriculture.

Improvement of gibberellin production by a newly isolated Fusarium fujikuroi mutant.

To obtain and investigate the potential mechanism for GA3 production in Fusarium fujikuroi GA-251, a high GA3 producer. Fusarium fujikuroi IMI 58289 was bred with Cobalt-60 (60 Co) radiation and lithium chloride treatment. The best mutant strain GA-251 was obtained for the subsequent optimization of fermentation conditions. The yield of GA3 by GA-251 was 2100mgl-1 , while the wild-type strain was 100mgl-1 , which is a 21-fold increase in the yield. To elucidate the mechanism of high GA3 yield of GA-251, the genome was sequenced and compared with wild-type strain IMI 58289. The results showed 2295 single nucleotide polymorphisms, 1242 small indels and 30 structural variants. These mutations were analysed and enriched in the MAPK signalling pathway, the mRNA surveillance pathway and endocytosis. The potential reasons for the improved GA3 biosynthesis were investigated. The potential mechanism of high GA3 yield was attributed to endocytosis pathway and histone modification proteins family. A mutant strain GA-251 in this work that could potentially be utilized in the industrial yield of GA3 . The comparative genome analysis would shed light onto the mechanism of yield improvement and be a theoretical guide for further metabolic engineering.

Flasher, a novel mutation in a glucosinolate modifying enzyme, conditions changes in plant architecture and hormone homeostasis.

Meristem function is underpinned by numerous genes that affect hormone levels, ultimately controlling phyllotaxy, the transition to flowering and general growth properties. Class I KNOX genes are major contributors to this process, promoting cytokinin biosynthesis but repressing gibberellin production to condition a replication competent state. We identified a suppressor mutant of the KNOX1 mutant brevipedicellus (bp) that we termed flasher (fsh), which promotes stem and pedicel elongation, suppresses early senescence, and negatively affects reproductive development. Map-based cloning and complementation tests revealed that fsh is due to an E40K change in the flavin monooxygenase GS-OX5, a gene encoding a glucosinolate (GSL) modifying enzyme. In vitro enzymatic assays revealed that fsh poorly converts substrate to product, yet the levels of several GSLs are higher in the suppressor line, implicating FSH in feedback control of GSL flux. FSH is expressed predominantly in the vasculature in patterns that do not significantly overlap those of BP, implying a non-cell autonomous mode of meristem control via one or more GSL metabolites. Hormone analyses revealed that cytokinin levels are low in bp, but fsh restores cytokinin levels to near normal by activating cytokinin biosynthesis genes. In addition, jasmonate levels in the fsh suppressor are significantly lower than in bp, which is likely due to elevated expression of JA inactivating genes. These observations suggest the involvement of the GSL pathway in generating one or more negative effectors of growth that influence inflorescence architecture and fecundity by altering the balance of hormonal regulators.

Neurosporaxanthin Overproduction by Fusarium fujikuroi and Evaluation of Its Antioxidant Properties.

Neurosporaxanthin (NX) is a carboxylic carotenoid produced by some filamentous fungi, including species of the genera Neurospora and Fusarium. NX biosynthetic genes and their regulation have been thoroughly investigated in Fusarium fujikuroi, an industrial fungus used for gibberellin production. In this species, carotenoid-overproducing mutants, affected in the regulatory gene carS, exhibit an upregulated expression of the NX pathway. Based on former data on a stimulatory effect of nitrogen starvation on carotenoid biosynthesis, we developed culture conditions with carS mutants allowing the production of deep-pigmented mycelia. With this method, we obtained samples with ca. 8 mg NX/g dry mass, in turn the highest concentration for this carotenoid described so far. NX-rich extracts obtained from these samples were used in parallel with carS-complemented NX-poor extracts obtained under the same conditions, to check the antioxidant properties of this carotenoid in in vitro assays. NX-rich extracts exhibited higher antioxidant capacity than NX-poor extracts, either when considering their quenching activity against [O2(1Δg)] in organic solvent (singlet oxygen absorption capacity (SOAC) assays) or their scavenging activity against different free radicals in aqueous solution and in liposomes. These results make NX a promising carotenoid as a possible feed or food additive, and encourage further studies on its chemical properties.

The Gibberellin Producer Fusarium fujikuroi: Methods and Technologies in the Current Toolkit.

In recent years, there has been a noticeable increase in research interests on the Fusarium species, which includes prevalent plant pathogens and human pathogens, common microbial food contaminants and industrial microbes. Taken the advantage of gibberellin synthesis, Fusarium fujikuroi succeed in being a prevalent plant pathogen. At the meanwhile, F. fujikuroi was utilized for industrial production of gibberellins, a group of extensively applied phytohormone. F. fujikuroi has been known for its outstanding performance in gibberellin production for almost 100 years. Research activities relate to this species has lasted for a very long period. The slow development in biological investigation of F. fujikuroi is largely due to the lack of efficient research technologies and molecular tools. During the past decade, technologies to analyze the molecular basis of host-pathogen interactions and metabolic regulations have been developed rapidly, especially on the aspects of genetic manipulation. At the meanwhile, the industrial fermentation technologies kept sustained development. In this article, we reviewed the currently available research tools/methods for F. fujikuroi research, focusing on the topics about genetic engineering and gibberellin production.

Research progress in biosynthesis and metabolism regulation of gibberellins in Gibberella fujikuroi

Gibberellin is one of the most important plant growth regulators, and widely used in agricultural production. However, the high cost of gibberellins production is restricting its efficient application. In recent years, biotechnological innovations have improved the synthesis of gibberellin. Gibberellin biosynthesis requires various enzymes. Current research focuses on the biosynthetic mechanisms of gibberellin and the metabolic engineering techniques to improve the production of gibberellin. This paper reviews the current research on gibberellin biosynthesis pathway, the key and enzymes environmental factors involved, and the metabolic regulation of gibberellin in Gibberella fujikuroi, summarizes the application of metabolic regulation in gibberellin biosynthesis, to provide the basis for achieving stable gibberellins production.

Different Phenotypes, Similar Genomes: Three Newly Sequenced Fusarium fujikuroi Strains Induce Different Symptoms in Rice Depending on Temperature.

Bakanae, caused by the hemibiotrophic fungus Fusarium fujikuroi, is one of the most important diseases of rice and is attributed to up to 75% of losses, depending on the strain and environmental conditions. Some strains cause elongation and thin leaves, whereas others induce stunting and chlorotic seedlings. Differences in symptoms are attributed to genetic differences in the strains. F. fujikuroi strains Augusto2, CSV1, and I1.3 were sequenced with Illumina MiSeq, and pathogenicity trials were conducted on rice cultivar Galileo, which is susceptible to bakanae. By performing gene prediction, single nucleotide polymorphism (SNP) calling, and structural variant analysis with a reference genome, we show how an extremely limited number of polymorphisms in genes not commonly associated with bakanae disease can cause strong differences in phenotype. CSV1 and Augusto2 were particularly close, with only 21,887 SNPs between them, but they differed in virulence, reaction to temperature, induced symptoms, colony morphology and color, growth speed, fumonisin, and gibberellin production. Genes potentially involved in the shift in phenotype were identified. Furthermore, we show how temperature variation may result in different symptoms even in rice plants inoculated with the same F. fujikuroi strain. Moreover, all of the F. fujikuroi strains became more virulent at higher temperatures. Significant differences were likewise observed in gibberellic acid production and in the expression of both fungal and plant gibberellin biosynthetic genes.

Isolation and Characterization of Phosphate Solubilizing Pseudomonas Species and Their Effect on Plant Growth Promotion

Plant growth-promoting rhizobacteria (PGPR) affect plant growth via various direct or indirect mechanisms. These microorganisms are the potential tools for sustainable agriculture. In this study, eight different Pseudomonas strains (PM1 - PM8) isolated from rhizospheric soil were characterized plant growth-promoting characteristics, including indole-3-acetic acid (IAA) and Gibberellin (GA3) production, phosphate solubilization, 1- aminocyclopropane-1-carboxylic acid (ACC) deaminase activity and effects of direct inoculation on plant growth parameters (fresh weight, shoot length). All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH4 - 5 at 2 days after inoculation. PM1, PM4, PM5 and PM7 species was capable of IAA production and PM4, PM5, PM7 and PM8 species have high activity for production of gibberellin (GA3) hormone. For ACC deaminase activity, PM1, PM3 and PM8 show high activity. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants for fresh weight and shoot length enhancement, so that these Pseudomonas species could be used as potential bio-fertilizer agents. Production of plant hormone IAA (A) and GA3 (B) from isolated Pseudomonas species.

Solid-state fermentation process for gibberellin production using enzymatic hydrolysate corn stalks

Solid-state fermentation was carried out for production of gibberellin via the addition of enzymatic hydrolysate from steam-exploded corn stalks during the culture period. The enzymatic hydrolysate from the steam-exploded corn stalks was added to the culture medium during the solid-state fermentation period, which improved gibberellin production. When the enzymatic hydrolysate was added into the 400 mL/kg dry basis substrate in the solid-state fermentation after 60 h, the temperature was 30 °C, the pH was 7.00, the mass ratio of solid to liquid was 1:1.1, and the fermentation period was 168 h. This led to the largest gibberellin yield (9.48 g/kg dry basis), and when compared with pre-optimization, the gibberellin yield increased by 135%. The optimum conditions to maximize the biomass for the fermentation process were obtained; the temperature was 32 °C for a gibberellin yield of 9.20 g/kg dry basis, the pH was 6.00 and the mass ratio of solid to liquid was 1:1.1 for a gibberellin yield of 9.48 g/kg dry basis, and the fermentation period was 96 h for a gibberellin yield of 6.94 g/kg dry basis. Therefore, a new alternative way for gibberellin production via solid-state fermentation has been demonstrated.

Gibberellin And IAA Production by Rhizobacteria From Various Private Forest

Rhizobacterium is a bacterium that colonizes plant roots, known to produce gibberellin and Indol Acetic Acid (IAA) to spur plant growth. This study aims to isolate rhizobacteria from various stands of community forest and to know the ability of rhizobacterial isolates to produce gibberellin and IAA. Research methods included rhizosphere soil sampling, bacterial isolation, gibberellin content test, and IAA. Results of rhizobacterial isolation were obtained by 35 isolates having a gibberellin production capability of about 1,959-4,322 mgl−1 and IAA about 0.389-2.370 mgl−1. The highest gibberellin production was obtained on the isolates of BB1, BB2 and BB3 while the highest IAA was in JS9 and JB4 isolates.

Effect of citronella essential oil fumigation on sprout suppression and quality of potato tubers during storage

Effect of citronella essential oil (CEO) fumigation on sprout suppression and quality of potato tubers during storage was investigated. Potato tubers were treated under conditions of single-phase (30 μL L−1, 0–10 d) and dual-phase (30 μL L−1, 0–10 d; 30 μL L−1, 35–90 d) fumigation. Changes in germination rate, weight loss, starch, reducing sugar, gibberellins (GA3), and α-solanine were measured. The results showed that CEO fumigation could control sprouting and improve the quality of potato tubers during storage compared to the non-treated tubers. CEO treatments inhibited the degradation of starch and the increase of reducing sugar content. The production of gibberellins (GA3) was suppressed, and the levels of α-solanine in the skin and flesh of potato tubers were decreased by CEO fumigation. Dual-phase CEO fumigation had a better effect on sprout suppression than single-phase fumigation, and possesses potential for postharvest application.

Production and Role of Hormones During Interaction of Fusarium Species With Maize (Zea mays L.) Seedlings.

It has long been known that hormones affect the interaction of a phytopathogen with its host plant. The pathogen can cause changes in plant hormone homeostasis directly by affecting biosynthesis or metabolism in the plant or by synthesizing and secreting the hormone itself. We previously demonstrated that pathogenic fungi of the Fusarium species complex are able to produce three major types of hormones: auxins, cytokinins, and gibberellins. In this work, we explore changes in the levels of these hormones in maize and mango plant tissues infected with Fusarium. The ability to produce individual phytohormones varies significantly across Fusarium species and such differences likely impact host specificity inducing the unique responses noted in planta during infection. For example, the production of gibberellins by F. fujikuroi leads to elongated rice stalks and the suppression of gibberellin biosynthesis in plant tissue. Although all Fusarium species are able to synthesize auxin, sometimes by multiple pathways, the ratio of its free form and conjugates in infected tissue is affected more than the total amount produced. The recently characterized unique pathway for cytokinin de novo synthesis in Fusarium appears silenced or non-functional in all studied species during plant infection. Despite this, a large increase in cytokinin levels was detected in F. mangiferae infected plants, caused likely by the up-regulation of plant genes responsible for their biosynthesis. Thus, the accumulation of active cytokinins may contribute to mango malformation of the reproductive organs upon infection of mango trees. Together, our findings provide insight into the complex role fungal and plant derived hormones play in the fungal–plant interactions.

Genetic Differentiation Associated with Fumonisin and Gibberellin Production in Japanese Fusarium fujikuroi.

Fusarium fujikuroi is a pathogenic fungus that infects rice. It produces several important mycotoxins, such as fumonisins. Fumonisin production has been detected in strains of maize, strawberry, and wheat, whereas it has not been detected in strains from rice seedlings infested with bakanae disease in Japan. We investigated the genetic relationships, pathogenicity, and resistance to a fungicide, thiophanate-methyl (TM), in 51 fumonisin-producing strains and 44 nonproducing strains. Phylogenetic analyses based on amplified fragment length polymorphism (AFLP) markers and two specific genes (a combined sequence of translation elongation factor 1α [TEF1α] and RNA polymerase II second-largest subunit [RPB2]) indicated differential clustering between the fumonisin-producing and -nonproducing strains. One of the AFLP markers, EATMCAY107, was specifically present in the fumonisin-producing strains. A specific single nucleotide polymorphism (SNP) between the fumonisin-producing and nonproducing strains was also detected in RPB2, in addition to an SNP previously found in TEF1α. Gibberellin production was higher in the nonproducing than in the producing strains according to an in vitro assay, and the nonproducing strains had the strongest pathogenicity with regard to rice seedlings. TM resistance was closely correlated with the cluster of fumonisin-nonproducing strains. The results indicate that intraspecific evolution in Japanese F. fujikuroi is associated with fumonisin production and pathogenicity. Two subgroups of Japanese F. fujikuroi, designated G group and F group, were distinguished based on phylogenetic differences and the high production of gibberellin and fumonisin, respectively.IMPORTANCEFusarium fujikuroi is a pathogenic fungus that causes rice bakanae disease. Historically, this pathogen has been known as Fusarium moniliforme, along with many other species based on a broad species concept. Gibberellin, which is currently known as a plant hormone, is a virulence factor of F. fujikuroi Fumonisin is a carcinogenic mycotoxin posing a serious threat to food and feed safety. Although it has been confirmed that F. fujikuroi produces gibberellin and fumonisin, production varies among strains, and individual production has been obscured by the traditional appellation of F. moniliforme, difficulties in species identification, and variation in the assays used to determine the production of these secondary metabolites. In this study, we discovered two phylogenetic subgroups associated with fumonisin and gibberellin production in Japanese F. fujikuroi.

Gibberellin production variability in <i>Fusarium fujikuroi</i> and its contributory factor

Gibberellin production variability in <i>Fusarium fujikuroi</i> and its contributory factor

Salt tolerance of Glycine max.L induced by endophytic fungus Aspergillus flavus CSH1, via regulating its endogenous hormones and antioxidative system

Abiotic stress resistance strategies are powerful approaches to sustainable agriculture because they reduce chemical input and enhance plant productivity. In current study, an endophytic fungus, Aspergillus flavus CHS1 was isolated from Chenopodium album Roots. CHS1 was initially screened for growth promoting activities like siderphore, phosphate solubilization, and the production of indole acetic acid and gibberellins and were further assayed for its ability to promote the growth of mutant Waito-C rice. The results revealed that different plant growth characteristic such as chlorophyll content, root-shoot length, and biomass production were significantly promoted during CHS1 treatment. This growth promotion action was due to the presence of various types of GAs and IAA in the endophyte culture filtrate. Significant up regulation with respect to levels in the control was observed in all endogenous plant GAs, after treatment with CHS1. Furthermore, to evaluate the potential of CHS1 against NaCl stress up to 400 mM, it was tested for its ability to improve soybean plant growth under NaCl stress. In endophyte-soybean interaction, CHS1 association significantly increased plant growth and attenuated the NaCl stress by down regulating ABA and JA synthesis. Similarly, it significantly elevated antioxidant activities of enzymes catalase, polyphenoloxidase, superoxide dismutase and peroxidase as compared to non-inoculated salt stress plants. Thus, CHS1 ameliorated the adverse effect of high NaCl stress and rescued soybean plant growth by regulating the endogenous plant hormones and antioxidative system. We conclude that CHS1 isolate could be exploited to increase salt resistant and yield in crop plants.

Evaluation of biological activities for salt-tolerant plant growth promoting rhizobacteria using different microbial carriers

Plant growth promoting rhizobacteria (PGPR) play an important role for improving plant growth and increasing productivity especially under stress condition. Fifty rhizobacterial isolates were isolated from the rhizosphere of the wheat plants. The rhizobacterial isolates were screened to select the most salt tolerant isolates. Then, the more tolerant isolates were evaluated for plant growth promoting activities such as ammonia, HCN, siderophores, nitrogenase, indole acetic acid (IAA) and gibberellins (GB) production as well as, root colonization ability, phosphate and potassium solubilization. The obtained data showed that, rhizobacterial isolate number (40) exhibited high records for most of PGPR activitiesand identified asPaenibacillus polymyxaMG309677.1using 16S rRNA gene sequencing techniques with 99% similarity. One of the problems which face production of microbial preparations for agriculture use is the maintenance of bacterial viability. In this respect, immobilization of P. Polymyxa MG309677.1 on three different carriers (peatmoss, compost and sawdust) was studied. Peatmoss and compost were used singly or combined with sawdust at two levels (50% and 75% sawdust).Bacterial populations, dehydrogenase activity, pH values and moisture contents were determined monthly up to six months of storage. Resultsrevealed that, using of peat moss either singly or combined with sawdust (50:50%) as a carrier gave the highest survival and population records forP. polymyxaMG309677.1even the end of storage period (180) days.

The dioxygenase GIM2 functions in seed germination by altering gibberellin production in Arabidopsis.

The phytohormones gibberellic acid (GA) and abscisic acid (ABA) antagonistically control seed germination. High levels of GA favor seed germination, whereas high levels of ABA hinder this process. The direct relationship between GA biosynthesis and seed germination ability need further investigation. Here, we identified the ABA-insensitive gain-of-function mutant germination insensitive to ABA mutant 2 (gim2) by screening a population of XVE T-DNA-tagged mutant lines. Based on two loss-of-function gim2-ko mutant lines, the disruption of GIM2 function caused a delay in seed germination. By contrast, upregulation of GIM2 accelerated seed germination, as observed in transgenic lines overexpressing GIM2 (OE). We detected a reduction in endogenous bioactive GA levels and an increase in endogenous ABA levels in the gim2-ko mutants compared to wild type. Conversely, the OE lines had increased endogenous bioactive GA levels and decreased endogenous ABA levels. The expression levels of a set of GA- and/or ABA-related genes were altered in both the gim2-ko mutants and the OE lines. We confirmed that GIM2 has dioxygenase activity using an in vitro enzyme assay, observing that GIM2 can oxidize GA12 . Hence, our characterization of GIM2 demonstrates that it plays a role in seed germination by affecting the GA metabolic pathway in Arabidopsis.

Preussia sp. BSL-10 producing nitric oxide, gibberellins, and indole acetic acid and improving rice plant growth

ABSTRACTPreussia sp. have been least known to improve plant growth and produce phytohormones. The current study investigated the production of nitric oxide (NO), indole-3-acetic acid (IAA), and gibberellins (GA4, GA7, GA15, and GA53) by a novel endophytic-fungal strain Preussia sp. BSL-10 using advanced chromatographic and spectroscopic techniques. Production of these phytohormones were validated by RT–PCR analysis, which indicated the expression of genes encoding tryptophan synthase (TRP), indole-3-acetamide hydrolase (IAAH), tryptophan-2-monooxygenase (IAAM), aldehyde dehydrogenase (ALD), GA4 desaturase (DES), geranylgeranyl-diphosphate synthase (GGS2), ent-desaturase oxidase (P450-4), GA14 synthase (P450-1) and nitrite reductase (NIRK/NIRS), cytochrome P450 (P450nor), nitrate reductase (NR), NOS-like (NOL), and nitric oxide reductase (QNOR/CNOR). In plant growth-promoting effects, the inoculation of Preussia sp. BSL-10 significantly increased the growth of dwarf mutant Waito-C and wild-type rice cultivars. In conclusion, utilizing new endophytic with the ability to produce NO, IAA, and gibberellins can be used to promote growth and yield of marginalized crops.

Aspergillus niger CSR3 regulates plant endogenous hormones and secondary metabolites by producing gibberellins and indoleacetic acid

ABSTRACTIn this study, an endophytic fungus, Aspergillus niger CSR3, was isolated from Cannabis sativa. The culture filtrate (CF) was initially screened for growth-promoting activities such as the presence of siderophores, phosphate solubilization, and the production of indole acetic acid (IAA) and gibberellins and was further assayed for its ability to promote the growth of mutant waito-C rice. Nearly all plant growth attributes examined (root–shoot length, biomass, and chlorophyll content) were significantly enhanced by treatment with CSR3. This growth promotion action was due to the presence of various types of gibberellins (GAs) and IAA in the endophyte CF. Moreover, the presence of GA pathway genes (P50-1, P450-3, P450-4, ggs2, and des) was confirmed by means of semi-quantitative RT-PCR. Finally, the application of CSR3 spore suspension with uniconazole and yucasin on maize seedlings revealed that, similar to exogenous IAA and GA3, CSR3 has the potential to alleviate the inhibitory effect of these inhibitors.