Exogenous Indole-3-acetic Acid Research Articles

Cytokinin-inducible response regulator SlRR6 controls plant height through gibberellin and auxin pathways in tomato.

Plant height is a key agronomic trait regulated by several phytohormones such as gibberellins (GAs) and auxin. However, little is known about how cytokinin (CK) participates in this process. Here, we report that SlRR6, a type-A response regulator in the CK signaling pathway, positively regulates plant height in tomato. SlRR6 was induced by exogenous kinetin and GA3, but inhibited by indole-3-acetic acid (IAA). Knock out of SlRR6 reduced tomato plant height through shortening internode length, while overexpression of SlRR6 caused taller plants due to increased internode number. Cytological observation of longitudinal stems showed that both knock out and overexpression of SlRR6 generated larger cells, but significantly reduced cell numbers in each internode. Further studies demonstrated that overexpression of SlRR6 enhanced GA accumulation and lowered IAA content, along with expression changes in GA- and IAA-related genes. Exogenous paclobutrazol and IAA treatments restored the increased plant height phenotype in SlRR6-overexpressing lines. Yeast two-hybrid, bimolecular fluorescence complementation, and co-immunoprecipitation assays showed that SlRR6 interacts with a small auxin up RNA protein, SlSAUR58. Moreover, SlSAUR58-overexpressing plants were dwarf with decreased internode length. Overall, our findings establish SlRR6 as a vital component in the CK signaling, GA, and IAA regulatory network that controls plant height.

Análise da ação exógena de auxinas e citocininas na superação da restrição radicular em ficus Benjamin verde e variegado

Abstract Green and variegated Benjamin fig (Ficus benjamina) often suffer from root restriction when grown in pots. While exogenous cytokinin applications have proven effective in reversing this stress, the possibility that exogenous auxins, either alone or in combination with cytokinin, may also be helpful has received little attention. In this work, we analyse the response of green and variegated Ficus benjamina rooted cuttings growing in small pots to exogenous supply of auxin and cytokinin at different concentrations, either in single or combined applications. Our results show that both benzyl aminopurine (BAP) and indole acetic acid (IAA), at the highest concentration tested (100 mg L-1) increased leaf development and plant biomass accumulation in green and variegated Ficus genotypes. However, exogenous IAA and BAP appeared to elicit differential plant morpho-physiological responses. While BAP tended to enhance leaf appearance more than IAA did, the latter promoted leaf expansion in a steadier manner than BAP, thus resulting in plants with less, but larger, leaves than those treated with cytokinin. Despite these differences in plant architecture, regression analysis suggests that hormonal-induced growth promotion was solely attributable to enhanced carbon assimilation. Rather unexpectedly, IAA promoted net assimilation and photosynthesis rates at least as effectively as cytokinin. Possible mechanisms involved in growth and development promotion by exogenous application of both hormones are discussed. Auxin treatment may help overcome root restriction in Ficus as effectively as cytokinin in terms of growth promotion, although differences in plant architecture may arise as compared with plants sprayed with the latter hormone.

Auxin Participates in the Regulation of the Antioxidant System in Malus baccata Borkh. Roots under Sub-Low Temperature by Exogenous Sucrose Application

Malus baccata Borkh., an apple rootstock, is found to be damaged by oxidation at sub-low root-zone temperature. In previous studies, we have found that exogenous sucrose could alleviate oxidative damage and increase the indole acetic acid (IAA) in roots under sub-low temperature (L). However, the role of IAA in sucrose-induced tolerance to L remains unclear. A pot experiment was conducted to evaluate the effects of exogenous sucrose and IAA synthesis/transport inhibitors (2,3,5-triiodobenzoic acid, TIBA; 4-biphenylboronic acid, BBo) on growth, IAA levels, sugars, and the antioxidant system of M. baccata under L. The results showed that the L treatment decreased IAA contents by 23.69% (48 h) and induced significant increases in root contents of malondialdehyde (MDA) and reactive oxygen (ROS), along with increasing catalase (CAT), ascorbate peroxidase (APX), and glucose-6-phosphate dehydrogenase (G6PDH) activities, while superoxide dismutase (SOD) and monodehydroascorbate reductase (MDHAR) activities first increased (24 h) and then decreased (48 h), and glutathione reductase (GR) and peroxidase (POD) activities significantly decreased. The L treatment also decreased ascorbate/oxidized ascorbate (AsA/DHA), glutathione/oxidized glutathione (GSH/GSSG), and coenzyme II/oxidized coenzyme II (NADPH/NADP+) ratios. Furthermore, the L treatment increased the contents of sucrose, fructose, glucose and sorbitol in the roots and suppressed plant growth. Sucrose pretreatment significantly increased IAA contents (12.42%, 24 h and 14.44%, 48 h) and decreased MDA and ROS contents, which improved the activities of antioxidant enzymes other than APX and increased the contents of AsA, GSH, and NADPH, and increased sucrose, fructose, and sorbitol contents and promoted plant growth. However, the sucrose + TIBA or BBo treatments decreased IAA contents and attenuated or almost abolished the positive effects of exogenous sucrose under sub-low temperature. Our findings indicate that IAA is involved in the sucrose-induced regulation of the antioxidant system in M. baccata roots under sub-low temperature and we provided theoretical references for further study on the adaptability of apple roots to low temperature.

Comparison of using plant growth promoting bacteria and exogenous indole acetic acid on rice under ozone stress

Plant growth promoting bacteria (PGPB) could mitigate plants under abiotic stress by influencing plant physiology and adaptation tolerance to stress. In this study, the aims are compared three strains of PGPB (Bacillus thuringensis, Lysinibacillus sp. N5 and Bacillus sp. N7) inoculation and both exogenous 5 μM and 10 μM IAA and compare the application of seed inoculation of PGPB and IAA and soil inoculation of PGPB/foliar spraying IAA on improving rice productivity under ozone stress. Ozone affected on rice productivity by decreasing AsA due to increasing malondialdehyde (MDA). Rice inoculated with PGPB and foliar spraying 5 μM IAA could lower MDA via enhancing IAA, AsA, and chlorophyll content. Nevertheless, improving rice productivity (grain yield and 100-grain weight) were found in both seed and soil inoculation of PGPB and applying seed inoculation of IAA5 and 10 μM. Therefore, both seed and soil inoculation of PGPB and seed inoculation of IAA were the best way for applying in rice under long-term ozone exposure.

Exogenous Indole-3-Acetic Acid Production and Phosphate Solubilization by Chlorella vulgaris Beijerinck in Heterotrophic Conditions

The role of soil-borne microalgae in the edaphic ecosystem is barely known, especially concerning their plant-growth-promoting traits, although they are used as biofertilizers. In this study, a microalgal strain isolated from soil cultivated with maize was evaluated as an exogenous producer of indole-3-acetic acid (IAA) in potato dextrose broth—PD—and sugarcane molasses, with or without tryptophan, and phosphate solubilizer (in ‘National Botanical Research Institute’s phosphate’—NBRIP—medium with tricalcium phosphate)with plant-growth-promoting traits, under heterotrophic conditions. The species of microalga was identified, by sequencing the ITS region in the rDNA and the morphological characteristics, as Chlorella vulgaris. Its growth was significantly higher in the PD medium, with slower growth in molasses. The addition of tryptophan did not influence the growth of C. vulgaris in either medium, but it increased the production of IAA, mainly in the PD medium, to 265 µg mL−1. The microalga grew in the medium with insoluble phosphate, releasing phosphorus into the medium (30 mg L−1 after 72 h). This is the first study on the application of C. vulgaris as a phosphate solubilizer. More studies should be performed on C. vulgaris as a prospective plant-growth-promoting microorganism, besides its ability to produce exogenous IAA, and further investigations should be conducted on developing inexpensive culture media.

Auxin alleviates cadmium toxicity by increasing vacuolar compartmentalization and decreasing long-distance translocation of cadmium in Poa pratensis

Kentucky bluegrass (Poa pratensis L.) hyperaccumulates cadmium (Cd) and exhibits a hypertolerance. Thus, it has potential for the phytoremediation of Cd-containing soil. Auxin signaling is involved in the response to Cd stress. However, the mechanisms of auxin-mediated detoxification and Cd translocation in plants remain unclear. This study aimed to investigate the effects of exogenous application of indole-3-acetic acid (IAA) on the Cd translocation, subcellular Cd distribution, chemical forms of Cd, and transcriptional regulation of Kentucky bluegrass. The results showed that the exogenous application of IAA increased the amount of organelle-bound Cd and vacuole-compartmentalized Cd in root cells, reduced the Cd concentration in the leaf tissues (epidermis, mesophyll, and vascular bundle) and root tissues (rhizodermis and cortex) but increased in the stele, and alleviate Cd-induced leaf chlorosis and growth inhibition. The expression of genes associated with Cd transporters (ABCs, ZIPs, NASs, OPTs, and YSLs), phosphatases, oxalate decarboxylases and lignin biosynthesis were significantly regulated by exogenous IAA under Cd stress. A positive regulation of phosphatases and oxalate decarboxylases genes related to an increase in phosphate- and oxalate-bound Cd, as well as a decrease in pectate- and protein-bound Cd and inorganic Cd, thereby contributing to a decrease in Cd phytotoxicity. The significant regulation of Cd transporters associated with decreasing the long-distance translocation of Cd, and the activation of lignin biosynthesis may contribute to the development of root endodermal barriers and increase the deposition of undissolved Cd phosphates and oxalate-bound Cd in the stele. These results revealed the important role of auxin in Cd detoxification and translocation in Kentucky bluegrass and they provide a theoretical basis for the phytoremediation of Cd-containing soil.

Genome-Wide Identification and Characterization of Auxin Response Factor (ARF) Gene Family Involved in Wood Formation and Response to Exogenous Hormone Treatment in Populus trichocarpa.

Auxin is a key regulator that virtually controls almost every aspect of plant growth and development throughout its life cycle. As the major components of auxin signaling, auxin response factors (ARFs) play crucial roles in various processes of plant growth and development. In this study, a total of 35 PtrARF genes were identified, and their phylogenetic relationships, chromosomal locations, synteny relationships, exon/intron structures, cis-elements, conserved motifs, and protein characteristics were systemically investigated. We also analyzed the expression patterns of these PtrARF genes and revealed that 16 of them, including PtrARF1, 3, 7, 11, 13-17, 21, 23, 26, 27, 29, 31, and 33, were preferentially expressed in primary stems, while 15 of them, including PtrARF2, 4, 6, 9, 10, 12, 18-20, 22, 24, 25, 28, 32, and 35, participated in different phases of wood formation. In addition, some PtrARF genes, with at least one cis-element related to indole-3-acetic acid (IAA) or abscisic acid (ABA) response, responded differently to exogenous IAA and ABA treatment, respectively. Three PtrARF proteins, namely PtrARF18, PtrARF23, and PtrARF29, selected from three classes, were characterized, and only PtrARF18 was a transcriptional self-activator localized in the nucleus. Moreover, Y2H and bimolecular fluorescence complementation (BiFC) assay demonstrated that PtrARF23 interacted with PtrIAA10 and PtrIAA28 in the nucleus, while PtrARF29 interacted with PtrIAA28 in the nucleus. Our results provided comprehensive information regarding the PtrARF gene family, which will lay some foundation for future research about PtrARF genes in tree development and growth, especially the wood formation, in response to cellular signaling and environmental cues.

Abscisic acid affects the floret numbers of inflorescence by regulating indole-3-acetic acid transport and accumulation in Lavandula angustifolia

Lavender flower essential oil had been used for a variety of therapeutic and cosmetic purposes. Previous studies mainly focused on essential oil composition and extraction methods, ignoring the factors that affected the production of essential oils, such as the floret number. This study aims to get a deep insight into the mechanism of floret number differences. Hormone analysis showed that there was a positive correlation between abscisic acid (ABA) content and the number of florets, while indole-3-acetic acid (IAA) was negatively correlated with floret numbers. RNA-Seq results showed that 1 035 differentially expressed genes (DEG) can be identified, of which 19 genes were significantly enriched in the carotenoid biosynthesis pathway. Common network analysis showed that the expression of the key genes of ABA pathway metabolism was negatively related to the DEG of IAA, especial IAA18. Exogenous IAA significantly inhibited the number of lavender florets and affected the expression of ABA metabolism genes. The results might be useful for a better understanding of the molecular mechanism of floret number differences.

CsIAGLU Regulates the Angle of Leaf Petiole by Affecting Endogenous Content of Auxin in Cucumber (Cucumis sativus L.).

The leaf angle is an important factor determining plant shoot architecture that may boost crop yield by increasing photosynthetic efficiency and facilitating high-density planting. Auxin is an important phytohormone involved in leaf angle regulation. Here, we identified two Single-Nucleotide Polymorphisms (SNPs) in the Indoleacetic Acid (IAA) glucosyltransferase gene CsIAGLU in 80 re-sequenced cucumber lines, of which the CsIAGLU717G,1234T is the dominant allele associated with a small leaf pedicle angle (LPA), whereas CsIAGLU717C,1234A is linked with a large LPA. CsIAGLU was highly expressed in leaves and petioles. In natural cucumber populations, the expression of CsIAGLU was negatively correlated with the LPA. The mutation of CsIAGLU induced by the CRISPR-Cas9 system resulted in elevated free IAA levels and enlarged cell expansion on the adaxial side of the petiole base, thus producing a greater LPA. Consistently, exogenous IAA treatment led to increased LPA and cell size. Therefore, our findings suggest that CsIAGLU functions as a negative regulator of LPA development via auxin-mediated cell expansion in cucumber, providing a valuable strategy for cucumber breeding with small LPAs.

Exogenous IAA improves the seedling growth of Syringa villosa via regulating the endogenous hormones and enhancing the photosynthesis

Auxin is a core regulator of plant growth and an effective promoter of cell division and cell expansion. Effects of auxin on improving plant growth of Syringa plants remain poorly elucidated. The present study was carried out to investigate the effects of exogenous indole-3-acetic acid (IAA) on morphogenesis, endogenous hormone content, photosynthesis capacity and find the basis of IAA-induced physiological effects of Syringa villosa by transcriptome sequencing technology. The results indicated that exogenous IAA significantly contributed to the plant height, stem diameter, leaf area and leaf number, which improved the accumulation of whole plant quality. Contents of endogenous IAA and gibberellic acid (GA3) were markedly higher than control after IAA treatment, while abscisic acid (ABA) content was significantly declined. For photosynthetic parameters, photosynthesis capacity was promoted by the application of IAA, as evidenced by a significant increase in net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) and a lower intercellular CO2 concentration (Ci). In addition, the transcriptome of S. villosa was first sequenced which revealed that genes related to the synthesis and signal transduction of growth-promoting hormones such as IAA, GA3, cytokinin (CTK), and brassinolide (BR) were significantly up-regulated within 2 h of IAA treatment. The expression levels of genes related to photosynthesis, antenna proteins, and carbon fixation of photosynthetic organisms were also enhanced by exogenous IAA. Additionally, the application of exogenous IAA had a positive effect on growth and development by rapidly inducing genes related to cell division and cell expansion. By profiling the transcriptome of IAA response, potential target genes were identified based on the hormone levels and photosynthesis. These results provide insights for further functional genomic studies on the regulatory mechanisms of IAA underlying Syringa plants architecture.

Widely targeted metabolomics analysis reveals the effect of exogenous auxin on postharvest resistance to Botrytis cinerea in kiwifruit (Actinidia chinensis L.)

Kiwifruit has high economic value but is susceptible to fungal infection after harvest. Botrytis cinerea (B. cinerea) is one of these fungal pathogens that cause huge economic losses and potential food safety problems. Auxin is a vital plant growth regulator that has been reported to play a role in plant defense. In this study, postharvest kiwifruit was treated with different concentrations of indole-3-acetic acid (IAA), after which the average incidence rate and lesion area were analyzed to determine the appropriate concentration of IAA for disease resistance. Defense enzymes, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were detected. Ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)-based widely targeted metabolomic analysis was used to investigate the mechanism of kiwifruit active compounds during IAA treatment and B. cinerea infection. Exogenous IAA treatment (especially at 50 μg/ mL) enhanced the resistance of kiwifruit to B. cinerea, resulting in reduced disease incidence and lesion area, and showed higher pathogen resistance-related defense enzyme activity in response to B. cinerea infection. Further metabolomic analysis revealed 776 metabolites in kiwifruit after IAA treatment, among which 51 metabolites in IAA_24h (24 h after IAA treatment) vs. CK (sterilized ddH2O treatment) fruit, 55 metabolites in IAA_72h (72 h after IAA treatment) vs. CK, and 67 metabolites in IAA_24h vs. IAA_72h were found to be significantly different. KEGG analysis showed that the resistance induced in kiwifruit by IAA treatment was closely related to the activation of phenylpropanoids, including flavonoids and phenols, terpenoids, carbohydrate metabolism, and hormone signaling pathways. These results might reveal the mechanism of IAA-induced resistance in kiwifruit and provide a new theoretical basis for the safe and efficient control of postharvest diseases in kiwifruit.

Priming with EDTA, IAA and Fe alleviates Pb toxicity in Trigonella Foneum graecum L. growth: Phytochemicals and secondary metabolites

This study evaluated the effects of the exogenous application of ethylenediaminetetraacetic acid (EDTA), indole-3-acetic acid (IAA) and iron sulfate (FeSO4) upon the phytochemical mechanisms of fenugreek grown under Pb-excess (2000 mg L−1 PbCl2). The results showed that chemical additives of EDTA and IAA as well as FeSO4 improved fenugreek germination parameters. The radicle length and the amylase activity were significantly improved under IAA treatment compared to EDTA and FeSO4. Exogenous FeSO4 was more effective to improving growth parameters. Moreover, the decrease in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was noted under all chemical additives especially under IAA application. In addition, it was more effective than EDTA and Fe in increasing catalase, glutathione (GSH), ascorbate peroxidase (APX), flavonoids and phenols while the increment superoxide dismutase (SOD) production was more pronounced under EDTA addition to Pb than other chelators. HPLC analysis revealed that the gallic was the major phenol produced under all chelators addition especially with IAA. In addition, the syringic acid was only produced with exogenous IAA while the quercetin was only detected under EDTA addition. Our results exhibited a higher IAA efficiency than EDTA and FeSO4 in mitigating Pb stress in fenugreek through up-regulated mechanisms of the antioxidant system for reducing reactive oxygen species (ROS) activities and enhancing special phenols.

Ameliorative effects of endogenous and exogenous indole-3-acetic acid on atrazine stressed paddy field cyanobacterial biofertilizer Cylindrospermum stagnale

Across the world, paddy fields naturally harbour cyanobacteria that function as biofertilizers and secrete various compounds like Indole-3-acetic acid (IAA) that help organisms in regulating their growth. Also, paddy field farming utilizes large amounts of pesticides (e.g. atrazine); but their continued application in the agricultural field causes toxicity in non-target cyanobacterial species that hinder their performance as a biofertilizer. Hence, the current study is an attempt to ameliorate the atrazine stress in cyanobacterium Cylindrospermum stagnale by addition of IAA (1 mM each) under different atrazine levels (0, 60, 80, 100, 120, 140 µg/l). Atrazine toxicity affected C. stagnale in a dose-dependent manner further experiments revealed that both the exogenous and endogenous IAA mitigated the detrimental effects of atrazine. It reduced MDA content and simultaneously increased chlorophyll content, total protein content, and multiple antioxidant enzyme activities [superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)] at 140 µg/l. A molecular docking study revealed that the pesticide binds to the D1 protein of the photoelectric chain in photosynthesis. Hence, the application of IAA or cyanobacterial biofertilizer that secretes a sufficient amount of IAA may assist sustainable agriculture in counteracting the atrazine toxicity.

Enhanced Cd-Accumulation in Typha latifolia by Interaction with Pseudomonas rhodesiae GRC140 under Axenic Hydroponic Conditions.

The Typha genus comprises plant species extensively studied for phytoremediation processes. Recently, Pseudomonas rhodesiae GRC140, an IAA-producing bacterium, was isolated from Typha latifolia roots. This bacterium stimulates the emergence of lateral roots of Arabidopsis thaliana in the presence and absence of cadmium. However, the bacterial influence on cadmium accumulation by the plant has not been determined. Moreover, the P. rhodesiae GRC140 effect in Cd phytoextraction by T. latifolia remains poorly understood. In this work, an axenic hydroponic culture of T. latifolia was established. The plants were used to evaluate the effects of cadmium stress in axenic plants and determine the effects of P. rhodesiae GRC140 and exogenous indole acetic acid (IAA) on Cd tolerance and Cd uptake by T. latifolia. Biomass production, total chlorophyll content, root electrolyte leakage, catalase activity, total glutathione, and Cd content were determined. The results showed that Cd reduces shoot biomass and increases total glutathione and Cd content in a dose-dependent manner in root tissues. Furthermore, P. rhodesiae GRC140 increased Cd translocation to the shoots, while IAA increased the Cd accumulation in plant roots, indicating that both treatments increase Cd removal by T. latifolia plants. These results indicate that axenic plants in hydroponic systems are adequate to evaluate the Cd effects in plants and suggest that T. latifolia phytoextraction abilities could be improved by P. rhodesiae GRC140 and exogenous IAA application.

Effect of indole-3-acetic acid supplementation on the physiology of Lolium perenne L. and microbial activity in cadmium-contaminated soil.

Cadmium (Cd) pollution has led to a serious deterioration in soil quality, plant growth, and human health. Therefore, restoration of soil quality is imperative. Phytoremediation is inexpensive and yields acceptable outcomes. Phytoremediation involves interaction between plant physiology and microbial activity and has been widely used in the remediation of Cd-contaminated soil. In the present study, Lolium perenne L. (perennial ryegrass) was planted in Cd-spiked soil and indole-3-acetic acid (IAA) was used to explore the physiological and biochemical characteristics of ryegrass as well as soil enzyme activity to remove Cd. The present study provides a theoretical basis for the phytoremediation of Cd-contaminated soil. The study investigated the effect of 30-mg/kg Cd-spiked soil on ryegrass (C) and 30-mg/kg Cd-spiked soil on ryegrass treated with 10-mg/kg IAA (CI) compared with uncontaminated soil and ryegrass as the control. At the end of the experiment, the ryegrass biomass, total chlorophyll, superoxide dismutase (SOD) activity, and soil invertase activity in C group were decreased by 33.7%, 23.0%, 29.7%, and 18.3%, respectively, whereas the peroxidase (POD) activity and soil basal respiration increased by 17.1% and 87.9%, respectively, compared with the control. In the CI group, the biomass of ryegrass, chlorophyll content, SOD activity, sucrase activity, fluorescein diacetate (FDA) hydrolase activity, and Cd removal rates increased by 14.5%, 19.9%, 24.3%, 12.1%, 20.4%, and 15.1%, respectively, whereas the POD activity, soil basal respiration, and Cd residues in the soil declined by 8.0%, 15.0%, and 17.0%, respectively, compared with the C group. Therefore, exposure to exogenous IAA alleviated the Cd stress on ryegrass and soil microorganisms and improved Cd absorption by ryegrass from the contaminated soil.

Determination of age-dependent endogenous indole-3 acetic acid (IAA) level in different organs of tomato plants

Indole-3-acetic acid (IAA), a phytohormone, is a crucial modulator of plant growth and development. As increased levels of endogenous IAA is responsible for inhibitory effects on plants, knowing its concentration would be helpful for controlling exogenous IAA input. An optimum endogenous IAA level is required for proper growth and development of plant with good health status. Hence, in this study, we have detected and measured IAA present in tomato root, shoot, leaves and fruits at two different ages (25 days and 50 days) by Reversed-Phase High Performance Liquid Chromatography (RP-HPLC), a suitable chromatographic method for separating molecules based on hydrophobicity. We observed that the endogenous level of IAA was significantly higher in roots, shoots, leaves and fruits at 50-days old plant compared to 25-days old mature plant (p-value <0.01). These results suggested that the IAA level may increase with the increase of the age of tomato plants. The tomato plants showed good vigor with IAA range 0.3-3.0 µg/g.F.W (25 days) and 3.4-7.5 µg/g.F.W (50 days) of age, respectively, implying that IAA might be used as a possible plant health indicator. Further study is needed to manipulate the IAA concentration for adequate plant growth and development.

Exogenous auxin-induced ENHANCER OF SHOOT REGENERATION 2 (ESR2) enhances femaleness of cucumber via activating CsACS2 gene.

Cucumber (Cucumis sativus L.) has been a model for the study of sex differentiation over the last two decades. Cucumber sex differentiation is mainly under genetic control, but plant growth regulators can also influence or even change it. However, the effect of exogenous auxin application on cucumber sex differentiation is not well understood at the physiological level. In this study, we explored the effects of different exogenous auxin concentrations on cucumber varieties with different mutant sex-controlling genotypes and found that there was a dosage effect of exogenous indole-3-acetic acid (IAA) on the enhancement of cucumber femaleness. Several ACC synthetase (ACS) family members responded directly to exogenous IAA, increasing endogenous ethylene synthesis, and this process appeared to be independent of the previously identified sex-related ACC oxidase CsACO2. We further demonstrated that ENHANCER OF SHOOT REGENERATION 2 (ESR2) responded to exogenous auxin induction by binding to ERE cis-acting element regions in the CsACS2 promoter, directly activating CsACS2 expression and thus increasing endogenous ethylene content, which may induce femaleness. These findings reveal that exogenous auxin increases cucumber femaleness by inducing a sex-controlling gene and promoting ethylene synthesis.

Transcription factor OsbZIP49 controls tiller angle and plant architecture through the induction of indole-3-acetic acid-amido synthetases in rice.

Tiller angle is an important determinant of plant architecture in rice (Oryza sativa L.). Auxins play a critical role in determining plant architecture; however, the underlying metabolic and signaling mechanisms are still largely unknown. In this study, we have identified a member of the bZIP family of TGA class transcription factors, OsbZIP49, that participates in the regulation of plant architecture and is specifically expressed in gravity-sensing tissues, including the shoot base, nodes and lamina joints. Transgenic rice plants overexpressing OsbZIP49 displayed a tiller-spreading phenotype with reduced plant height and internode lengths. In contrast, CRISPR/Cas9-mediated knockout of OsbZIP49 resulted in a compact architecture. Follow-up studies indicated that the effects of OsbZIP49 on tiller angles are mediated through changes in shoot gravitropic responses. Additionally, we provide evidence that OsbZIP49 activates the expression of indole-3-acetic acid-amido synthetases OsGH3-2 and OsGH3-13 by directly binding to TGACG motifs located within the promoters of both genes. Increased GH3-catalyzed conjugation of indole-3-acetic acid (IAA) in rice transformants overexpressing OsbZIP49 resulted in the increased accumulation of IAA-Asp and IAA-Glu, and a reduction in local free auxin, tryptamine and IAA-Glc levels. Exogenous IAA or naphthylacetic acid (NAA) partially restored shoot gravitropic responses in OsbZIP49-overexpressing plants. Knockout of OsbZIP49 led to reduced expression of both OsGH3-2 and OsGH3-13 within the shoot base, and increased accumulation of IAA and increased OsIAA20 expression levels were observed in transformants following gravistimulation. Taken together, the present results reveal the role transcription factor OsbZIP49 plays in determining plant architecture, primarily due to its influence on local auxin homeostasis.

HOMEOBOX PROTEIN 24 mediates the conversion of indole‐3‐butyric acid to indole‐3‐acetic acid to promote root hair elongation

Indole-3-acetic acid (IAA) is a predominant form of active auxin in plants. In addition to de novo biosynthesis and release from its conjugate forms, IAA can be converted from its precursor indole-3-butyric acid (IBA). The IBA-derived IAA may help drive root hair elongation in Arabidopsis thaliana seedlings, but how the IBA-to-IAA conversion is regulated and affects IAA function requires further investigation. In this study, HOMEOBOX PROTEIN 24 (HB24), a transcription factor in the zinc finger-homeodomain family (ZF-HD family) of proteins, was identified. With loss of HB24 function, defective growth occurred in root hairs. INDOLE-3-BUTYRIC ACID RESPONSE 1 (IBR1), which encodes an enzyme involved in the IBA-to-IAA conversion, was identified as a direct target of HB24 for the control of root hair elongation. The exogenous IAA or auxin analogue 1-naphthalene acetic acid (NAA) both rescued the root hair growth phenotype of hb24 mutants, but IBA did not, suggesting a role for HB24 in the IBA-to-IAA conversion. Therefore, HB24 participates in root hair elongation by upregulating the expression of IBR1 and subsequently promoting the IBA-to-IAA conversion. Moreover, IAA also elevated the expression of HB24, suggesting a feedback loop is involved in IBA-to-IAA conversion-mediated root hair elongation.

Key auxin response factor (ARF) genes constraining wheat tillering of mutant dmc.

Tillering ability is a key agronomy trait for wheat (Triticum aestivum L.) production. Studies on a dwarf monoculm wheat mutant (dmc) showed that ARF11 played an important role in tillering of wheat. In this study, a total of 67 ARF family members were identified and clustered to two main classes with four subgroups based on their protein structures. The promoter regions of T. aestivum ARF (TaARF) genes contain a large number of cis-acting elements closely related to plant growth and development, and hormone response. The segmental duplication events occurred commonly and played a major role in the expansion of TaARFs. The gene collinearity degrees of the ARFs between wheat and other grasses, rice and maize, were significantly high. The evolution distances among TaARFs determine their expression profiles, such as homoeologous genes have similar expression profiles, like TaARF4-3A-1, TaARF4-3A-2 and their homoeologous genes. The expression profiles of TaARFs in various tissues or organs indicated TaARF3, TaARF4, TaARF9 and TaARF22 and their homoeologous genes played basic roles during wheat development. TaARF4, TaARF9, TaARF12, TaARF15, TaARF17, TaARF21, TaARF25 and their homoeologous genes probably played basic roles in tiller development. qRT-PCR analyses of 20 representative TaARF genes revealed that the abnormal expressions of TaARF11 and TaARF14 were major causes constraining the tillering of dmc. Indole-3-acetic acid (IAA) contents in dmc were significantly less than that in Guomai 301 at key tillering stages. Exogenous IAA application significantly promoted wheat tillering, and affected the transcriptions of TaARFs. These data suggested that TaARFs as well as IAA signaling were involved in controlling wheat tillering. This study provided valuable clues for functional characterization of ARFs in wheat.