Auxin-like Compounds Research Articles

Anti-oomycete activity and plant growth promoting properties of avocado fungal endophytes

Fungal endophytes are known as promising plant growth-promoting microorganisms. Surprisingly, despite the economic importance of avocado, the antimicrobial and plant growth-promoting properties of its fungal endophytes have seldom been investigated. Our objectives were to evaluate the anti-oomycete activity of avocado fungal endophytes and assess their potential plant growth-promoting properties, using Arabidopsis thaliana as a model plant. In total, 89 fungal isolates were obtained from the roots of avocado trees and grouped into 24 morphotypes. One isolate per morphotype was randomly selected to assess its antagonistic activity against Phytophthora cinnamomi through dual culture assays. The strongest inhibition of P. cinnamomi was induced by endophytic fungi belonging to the Fusarium, Mortierella and Penicillium genera. The six most promising fungal isolates were selected to assess their plant growth-promoting traits in co-inoculation assays with A. thaliana. All tested fungal endophytes were able to modify the plant root architecture and increase the number of lateral roots. Moreover, an accumulation of auxins was detected in the xylem and meristematic zone of plants inoculated with Mortierella sp. PC-T3-3.1 and Metapochonia sp. PC-T2-4.2, whilst auxin accumulation was restricted to the emerging lateral roots of plants inoculated with Penicillium sp. C-T0-3.1. Indole quantification showed that Penicillium sp. C-T0-3.1 produced the highest concentration of indole acetic acid (IAA) and indole butyric acid (IBA), despite the lack of auxin responsiveness in plants in the co-inoculation assays. Mortierella sp. PC-T3-3.1 and Metapochonia sp. PC-T2-4.2 produced more IAA and IBA when co-inoculated with A. thaliana than when growing alone, which suggests that sensing of plant signals could induce the production of auxin-like compounds by these two endophytes. Collectively, our findings contribute to elucidate the mechanisms underpinning the plant growth-promoting activity of avocado fungal endophytes, which will be key to harness their full metabolic potential.

Peribacillus frigoritolerans T7-IITJ, a potential biofertilizer, induces plant growth-promoting genes of Arabidopsis thaliana.

This study aimed to isolate plant growth and drought tolerance-promoting bacteria from the nutrient-poor rhizosphere soil of Thar desert plants and unravel their molecular mechanisms of plant growth promotion. Among our rhizobacterial isolates, Enterobacter cloacae C1P-IITJ, Kalamiella piersonii J4-IITJ, and Peribacillus frigoritolerans T7-IITJ, significantly enhanced root and shoot growth (4-5-fold) in Arabidopsis thaliana under PEG-induced drought stress. Whole genome sequencing and biochemical analyses of the non-pathogenic bacterium T7-IITJ revealed its plant growth-promoting traits, viz., solubilization of phosphate (40-73µg/ml), iron (24±0.58mm halo on chrome azurol S media), and nitrate (1.58±0.01µg/ml nitrite), along with production of exopolysaccharides (125±20µg/ml) and auxin-like compounds (42.6±0.05µg/ml). Transcriptome analysis of A. thaliana inoculated with T7-IITJ and exposure to drought revealed the induction of 445 plant genes (log2fold-change>1, FDR<0.05) for photosynthesis, auxin and jasmonate signalling, nutrient uptake, redox homeostasis, and secondary metabolite biosynthesis pathways related to beneficial bacteria-plant interaction, but repression of 503 genes (log2fold-change < -1) including many stress-responsive genes. T7-IITJ enhanced proline 2.5-fold, chlorophyll 2.5-2.8-fold, iron 2-fold, phosphate 1.6-fold, and nitrogen 4-fold, and reduced reactive oxygen species 2-4.7-fold in plant tissues under drought. T7-IITJ also improved the germination and seedling growth of Tephrosia purpurea, Triticum aestivum, and Setaria italica under drought and inhibited the growth of two plant pathogenic fungi, Fusarium oxysporum, and Rhizoctonia solani. P. frigoritolerans T7-IITJ is a potent biofertilizer that regulates plant genes to promote growth and drought tolerance.

The Application of Auxin-like Compounds Promotes Cold Acclimation in the Oilseed Rape Plant.

Cold is a major environmental key factor influencing plant growth, development, and productivity. Responses and adaption processes depend on plant physiological and biochemical modifications, first of all via the hormonal system. Indole-3-acetic acid (IAA) plays a critical role in the processes of plant functioning. To assess the influence of the auxin-like compounds 1-[2-chloroethoxycarbonylmethyl]-4-naphthalenesulfonic acid calcium salt (TA-12) and 1-[2-dimethylaminoethoxycarbonylmethyl]naphthalene chloromethylate (TA-14) in the process of cold acclimation, long-term field trials over four years were performed with two rapeseed (Brassica napus L.) plant cultivars with different wintering resistance in temperate-zone countries. In these two rapeseed cultivars, namely ‘Casino’ (less resistant) and ‘Valesca’ (more resistant), investigations were conducted in the terminal buds and root collars. The application of auxin-like compounds revealed a close interlinkage between the composition of dehydrins and the participation of the phytohormone IAA in the adaptation processes. By applying TA-12 and TA-14, the importance of the proteins, especially the composition of the dehydrins, the IAA amount, and the status of the oilseed rape cultivars at the end of the cold acclimation period were confirmed. Following on from this, when introducing oilseed rape cultivars from foreign countries, it may also be of value to assess their suitability for cultivation in temperate-zone countries.

Marine seaweeds (biofertilizer) significance in sustainable agricultural activities: A review

The marine environment provides adaptive conditions for many organisms, such as algae, shellfish, mussels, sponges, corals, and barnacles. There is a range of algae in Pakistan’s coastal waters, but, unfortunately, they are not used and are wasted. Efficient use of a valuable natural resource that can increase the efficiency of agricultural operations. Algae are abundant and easily found in a wet environment, either in seawater or freshwater. The seaweed and its products are extensively used in crops cultivation to enhance production systems due to many plant-growth-promoting compounds. It has been traditionally used for the fertilization of the fields since the ancient world. Some seaweeds and auxins, a plant hormone responsible for the vegetative growth and auxin-like compounds, have been found. Algae and their associated compounds can support the onset of seed germination and plant root growth, which applies to temperature, resistance to abiotic stress, and improves plants’ ability to absorb nutrients.

Evaluation and selection of culture media for the detection of auxin-like compounds and phosphate solubilization on soil yeasts

The detection of characteristics associated with plant growth promotion has been studied frequently on bacteria and some of these detection methods were also used on yeasts. Sometimes, these methods, designed for prokaryotes, were used with no prior analysis regarding growth and production on eukaryotes. The aim of the present study was to assess and select the best suitable media for the detection of auxin-like compound production and inorganic-P solubilization on yeast strains. Already published media and new formulations, as well as yeasts with different genetic backgrounds were used for the comparison. Media were selected based on the adequate growth of yeast strains and reliable recognition of the studied features either by an easy detection of the metabolite (color or halo production) or simple medium preparation (low number of reagent and regular autoclaving). We propose here the use of the new DEV medium with glucose and with tryptophan to identify auxin-like compounds producers; and CPM medium (a variation from Custer's Chalk Medium) as base medium to identify yeasts capable of inorganic-P solubilization.

Effects of 2,4-Dichlorophenoxyacetic Acid on Cucumber Fruit Development and Metabolism.

The auxin-like compound 2,4-dichlorophenoxyacetic acid (2,4-D) has been widely used as a plant growth regulator in cucumber fruit production; however, its influence on fruit development and metabolism has not been evaluated. In this study, the phenotype of cucumber fruits in both 2,4-D treatment and non-treatment control groups were recorded, and the metabolome of different segments of cucumber fruit at various sampling time points were profiled by a standardized non-targeted metabolomics method based on UPLC-qTOF-MS. The application of 2,4-D increased the early growth rate of the fruit length but had no significant effect on the final fruit length, and produced cucumber fruits with fresh flowers at the top. The 2,4-D treatment also affected the cucumber fruit metabolome, causing significant changes in the stylar end at 4 days after flowering (DAF). The significantly changed metabolites were mainly involved in methionine metabolism, the citric acid cycle and flavonoid metabolism pathways. At the harvest stage, 2,4–D treatment significantly decreased the levels of flavonoids and cinnamic acid derivatives while increased the levels of some of the amino acids. In summary, exogenous application of 2,4-D can greatly alter the phenotype and metabolism of cucumber fruit. These findings will assist in exploring the mechanisms of how 2,4-D treatment changes the fruit phenotype and evaluating the influence of 2,4-D treatment on the nutritional qualities of cucumber fruit.

Auxin-like compounds act as protectors against UV-B irradiation in garden pea plants

AbstractPretreatment with the original auxin physiological analogues 1-[2-chloroethoxycarbonylmethyl]-4-naphthalenesulfonic acid calcium salt (TA-12) and 1-[2-dimethylaminoethoxicarbonylmethyl]naphthalene chlormethylate (TA-14) and subsequent UV-B irradiation (180 min at λmax 312 nm for 6.6 kJ·m−2) of pea plants (Pisum sativum L.) was investigated to assess if foliar application of these compounds has ability to attenuate the negative effects caused by UV-B stress. UV-B treatment increased malondialdehyde (MDA) and proline levels as well as superoxide dismutase, catalase and guaiacol peroxidase activities, but decreased hydrogen peroxide, low-molecular thiols, total phenolics and total soluble protein contents. The pre-treatment with TA compounds decreased the oxidative stress provoked by UV-B radiation detected by lower level of MDA, increased the content of thiols and UV-absorbing compounds and had favourable effect on H2O2 content and enzymatic activities. Exogenous application of auxin-like compounds on pea plantlets successfully counteracted UV-B induced oxidative stress via activation of ROS detoxifying enzymes and non-enzymatic antioxidants.

Screening of cultivable endophytic bacterial isolates for their plant growth promoting activity in rice

Large amounts of chemical fertilizers are being used to enhance the growth and development of the rice (Oryza sativa L.) crop. Chemical fertilizers are too expensive; hence, endophytic bacteria with growth promoting activity can be considered to promote the crop plant growth to minimize the usage of chemical fertilizers and to have sustainable rice cultivation. This study was undertaken to explore the potential of endophytic bacterial isolates (EBIs) which were isolated previously. The objective of this study was to evaluate 800 EBIs for auxin-like activity and its effect on early seedlings growth of rice. Surface sterilized rice seeds were treated separately with different EBIs and allowed to grow in Petriplates (containing sterile cotton) for 10 days. On the 10th day, the root length of the rice seedlings was measured and recorded to determine the auxin-like activity of EBIs. Results showed that 9 different isolates out of 800 EBIs screened to have significant effect on rice root and shoot growth when compared to control. The increase in root length was in the range of 58.30% to 81.68%, in comparison to root length of seedlings from control category. Based on the results, it was concluded that the positive root growth promoting response of rice seedlings could be due to the production of auxin-like compounds by the EBIs. These research findings could be useful in developing endophytic bacteria based formulation to enhance the growth of rice crop.

Fungal Laccase-Catalyzed Oxidation of Naturally Occurring Phenols for Enhanced Germination and Salt Tolerance of Arabidopsis thaliana: A Green Route for Synthesizing Humic-like Fertilizers.

Fungal laccases have been highlighted as a catalytic tool for transforming phenols. Here we demonstrate that fungal laccase-catalyzed oxidations can transform naturally occurring phenols into plant fertilizers with properties very similar to those of commercial humic acids. Treatments of Arabidopsis thaliana with highly cross-linked polyphenolic products obtained from a mixture of catechol and vanillic acid were able to enhance the germination and salt tolerance of this plant. These results revealed that humic-like organic fertilizers can be produced via in vitro enzymatic oxidation reactions. In particular, the root elongation pattern resulting from the laccase products was comparable to that resulting from an auxin-like compound. A detailed structural comparison of the phenol variants and commercial humic acids revealed their similarities and differences. Analyses based on SEM, EFM, ERP, and zeta-potential measurement showed that they both formed globular granules bearing various hydrophilic/polar groups in aqueous and solid conditions. Solid-phase 13C NMR, FT-IR-ATR, and elemental analyses showed that more nitrogen-based functional and aliphatic groups were present in the commercial humic acids. Significant differences were also identifiable with respect to particle size and specific surface area. High-resolution (15 T) FT-ICR mass spectrometry-based van Krevelen diagrams showed the compositional features of the variants to be a subset of those of the humic acids. Overall, our study unraveled essential structural features of polyaromatics that affect the growth of plants, and also provided novel bottom-up ecofriendly and finely tunable pathways for synthesizing humic-like fertilizers.

Detection of plant growth enhancing features in psychrotolerant yeasts from Patagonia (Argentina).

This study explores the biotechnological potential for plant production of twelve psychrotolerant yeasts strains from Northwest-Patagonia. These strains were isolated from different substrates associated with Nothofagus sp. in native forests and Vaccinium sp. in a commercial plantation. Yeasts characterization was performed using in vitro assays to evaluate the production of auxin-like compounds and siderophores, ability to solubilize inorganic phosphate and to reduce common plant pathogen growth. Strain YF8.3 identified as Aureobasidium pullullans was the main producer of auxin-like and siderophores compounds. Phosphate solubilization was a characteristic observed by strains L8.12 and CRUB1775 identified as Holtermaniella takashimae and Candida maritima, respectively. Different yeast strains were able to inhibit the growth of Verticillium dahliae PPRI5569 and Pythium aphanidermatum PPRI 9009, but they all failed to inhibit the growth of Fusarium oxysporum PPRI5457. The present study, suggests that yeasts present in different environments in Northwestern-Patagonian have physiological in vitro features which may influence plant growth. These results are promising for the developing of biological products based on Patagonian yeasts for plant production in cold-temperate regions.

Ecological functions of Trichoderma spp. and their secondary metabolites in the rhizosphere: interactions with plants.

Trichodermaspp. are common soil and root inhabitants that have been widely studied due to their capacity to produce antibiotics, parasitize other fungi and compete with deleterious plant microorganisms. These fungi produce a number of secondary metabolites such as non-ribosomal peptides, terpenoids, pyrones and indolic-derived compounds. In the rhizosphere, the exchange and recognition of signaling molecules byTrichodermaand plants may alter physiological and biochemical aspects in both. For example, severalTrichodermastrains induce root branching and increase shoot biomass as a consequence of cell division, expansion and differentiation by the presence of fungal auxin-like compounds. Furthermore,Trichoderma, in association with plant roots, can trigger systemic resistance and improve plant nutrient uptake. The present review describes the most recent advances in understanding the ecological functions ofTrichodermaspp. in the rhizosphere at biochemical and molecular levels with special emphasis on their associations with plants. Finally, through a synthesis of the current body of work, we present potential future research directions on studies related toTrichodermaspp. and their secondary metabolites in agroecosystems.

Effects of humic acids derived from lignite and cattle manure on antioxidant enzymatic activities of barley root

ABSTRACTTo investigate the effects of humic acids (HAs) on the ability of plants to defend themselves against oxidative stress, barley was hydroponically cultured in the absence and presence of HAs, and the antioxidant enzymatic activities (catalase, superoxide dismutase, ascorbate and glutathione peroxidases) of root tissue were evaluated. Auxin-like structures in HAs, which were extracted from an oxidation product of lignite (LHA) and compost derived from cattle manure (CHA), were identified by pyrolysis-gas chromatography/mass spectrometry (GC/MS) with tetramethylammonium hydroxide. The LHA, which had the lower molecular weight, was more effective in promoting the growth of barley root than CHA. However, the amounts of auxin-like structures in the CHA were much higher than those for LHA. The antioxidant enzymatic activities were initially decreased in the presence of LHA and CHA at the first day after refreshing the culture solution, but were significantly increased on the second day. The CHA sample, which contained relatively high levels of phenolic acids that contained auxin-like structures, was effective in increasing four types of enzymatic activities, while the activities of catalase and ascorbate peroxidase were increased in the presence of LHA, which contains naphthalene derivatives. These results indicate that using HAs as a supplement can be effective in enhancing antioxidation enzymatic activities, while the appearance of the effects is retarded because of the decomposition and release of auxin-like compounds from HAs by organic acids from the plant roots.

Co-inoculation of Glomus intraradices andTrichoderma atroviride acts as a biostimulant topromote growth, yield andnutrient uptake ofvegetable crops.

The application of beneficial microorganisms at transplanting can promote rapid transplant establishment (starter effect) for achieving early and high yields. The aim of this study was to evaluate the biostimulant effects of Glomus intraradices BEG72 (G) and Trichoderma atroviride MUCL 45632 (T) alone or in combination on plant growth parameters, yield, chlorophyll index (SPAD), chlorophyll fluorescence and mineral composition of several vegetable crops. The T. atroviride strain was capable of producing siderophores and auxin-like compounds under a wide range of substrate pH conditions (5.5-8.0). The highest shoot, root dry weight, SPAD and chlorophyll fluorescence in lettuce, tomato and zucchini was observed in the G + T combination, followed by a single inoculation of G or T, whereas the lowest values were recorded in the uninoculated plants. Under greenhouse conditions, the shoot dry weight was significantly increased by 167%, 56%, 115%, 68% and 58% in lettuce, melon, pepper, tomato and zucchini, respectively, when supplied with both beneficial microorganisms in comparison with the control. This increase in root and shoot weight was associated with an increased level of nutrient uptake (e.g. P, Mg, Fe, Zn and B). Under open field conditions, the lettuce shoot and root dry weight increased by 61% and 57%, respectively, with biostimulant microorganism application in field conditions. For zucchini, early and total yields were significantly increased by 59% and 15%, respectively, when plants were inoculated with both microorganisms. The application of the biostimulant tablet containing both G and T can promote transplant establishment and vegetable crop productivity in a sustainable way.

Morfología y bioactividad de la cianobacteria Leptolyngbya en los campos de cultivo de arroz

The genus Leptolyngbya comprises filamentous cyanobacteria that are important in rice fields. In the rhizosphere, cyanobacteria produce a variety of secondary metabolites such as auxins that are important in agriculture soil performance. To assess this, Leptolyngbya strain MMG-1, was isolated from the rhizosphere of rice plants and described. For this, the morphology of this strain was studied by light microscopy as well as by confocal laser scanning microscopy. Besides, the ability of this strain to synthesize an auxin-like bioactive com- pound was demonstrated under various culture conditions (different amounts of tryptophan; pH; different alter- nating light:dark periods; duration of the incubation). The auxin-like compound was extracted from the culture of Leptolyngbya strain MMG-1 and identified as indole-3-acetic acid (IAA) by thin layer chromatography (TLC) as well as by high performance liquid chromatography (HPLC). Our results showed that the strain required the precursor L-tryptophan for the synthesis of IAA. Leptolyngbya strain MMG-1 accumulated IAA intracellularly. The IAA secreted by Leptolyngbya strain MMG-1 was significantly correlated with the initial concentration of L-tryptophan in the medium, as well as with the duration of the incubation. The bioactivity of the secreted IAA was determined by its effect on the rooting pattern of Pisum sativum seedlings. The culture supernatant of Leptolyngbya strain MMG-1 stimulated the seedling lateral rooting, while it decreased root length. Hence, rhizospheric Leptolyngbya produced auxin under different conditions and affected the plants rooting pattern.

Changes in auxin level in the course of growth of a sunflower crown-gall suspension culture

The auxin level in the cell mass and culture medium was determined by means of the &lt;em&gt;Avena&lt;/em&gt; straight caleoptile test in various periods of the suspension culture cycle of the sunflower crown-gall tumour. The investigations were performed in the course of the zero passage (PO) and first one (Pl), differing in their time of duration of maximum growth and its intensity. In both passages the intra- and extra-cellular auxin levels reach values of the same order. At the beginning of the maximal growth phase the activity corresponding to IAA in the cells prevails over that of the other auxin-like compounds. This disproportion diminishes with further development of the culture, and with the beginning of the stationary phase the cellular IAA level is lower than that of the remaining auxin-like compounds. The short phase of maximal growth (PO) occurs with an auxin level decreasing in the cell mass and increasing in the medium, and towards the end of the cycle these levels become equal. During the long phase of maximal growth (Pl) the total amount of auxins in the cells increases and is 2-3 times higher than in the medium, whereas IAA in the cells remains at a constant level. These results suggest that the participation of IAA in the intracellular pool of auxin-like substances is decisive for the mitotic activity of the cells and maintenance of growth in the culture.

Influence of Pythium oligandrum Biocontrol on Fungal and Oomycete Population Dynamics in the Rhizosphere

Fungal and oomycete populations and their dynamics were investigated following the introduction of the biocontrol agent Pythium oligandrum into the rhizosphere of tomato plants grown in soilless culture. Three strains of P. oligandrum were selected on the basis of their ability to form oospores (resting structures) and to produce tryptamine (an auxin-like compound) and oligandrin (a glycoprotein elicitor). Real-time PCR and plate counting demonstrated the persistence of large amounts of the antagonistic oomycete in the rhizosphere throughout the cropping season (April to September). Inter-simple-sequence-repeat analysis of the P. oligandrum strains collected from root samples at the end of the cropping season showed that among the three strains used for inoculation, the one producing the smallest amount of oospores was detected at 90%. Single-strand conformational polymorphism analysis revealed increases in the number of members and the complexity of the fungal community over time. There were no significant differences between the microbial ecosystems inoculated with P. oligandrum and those that were not treated, except for a reduction of Pythium dissotocum (ubiquitous tomato root minor pathogen) populations in inoculated systems during the last 3 months of culture. These findings raise interesting issues concerning the use of P. oligandrum strains producing elicitor and auxin molecules for plant protection and the development of biocontrol.

Physiological characterization and electron microscopic investigation of cyanobacteria associated with wheat rhizosphere

Physiological attributes of a set of cyanobacterial strains, isolated from the rhizosphere of wheat (var. HD 2687), identified as belonging to the genera Calothrix (n = 3), Westiellopsis (1), Hapalosiphon (2) and Nostoc (2), were axenized and evaluated. The concentrated culture filtrates of three cyanobacterial strains - C. ghosei, H. intricatus and Nostoc sp. were able to enhance germination percentage, radicle and coleoptile length in inhibition experiments with wheat seeds. Indole-3-acetic acid (IAA) production was recorded in light and dark (+0.5 % glucose) incubated cultures. Incubation in the presence of tryptophan significantly enhanced IAA production. Acetylene-reducing activity was higher in light incubated cultures of Nostoc sp. followed by C. ghosei, while in the dark, C. ghosei recorded highest values. TLC of the filtrates revealed the presence of several amino acids such as histidine, and auxin-like compounds. Co-culturing with selected strains recorded significant enhancement in plant chlorophyll. Root sections of wheat seedlings co-cultured with C. ghosei revealed the presence of short filaments inside the root hairs and cortical region. Such strains can be promising candidates for developing plant growth promoting associations for wheat crop, besides serving as model systems for understanding the metabolic interactions of cyanobacteria with host plant, such as wheat.

Auxin-Induced Resistance to Common Scab Disease of Potato Linked to Inhibition of Thaxtomin A Toxicity.

Production of the phytotoxin thaxtomin A by pathogenic Streptomyces spp. is essential for induction of common scab disease in potato. Prior studies have shown that foliar application of sublethal concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and other auxin or auxin-like compounds significantly reduced severity and occurrence of common scab in subsequently produced tubers. However, the means of disease suppression by these compounds was not known. We confirm the disease suppressive activity of 2,4-D. Detailed tuber physiological examination showed that lenticel numbers, lenticel external dimensions, and periderm thickness and structure, physiological features believed to be critical to Streptomyces scabiei infection, were not substantially changed by 2,4-D treatments, negating a possible mechanism for disease suppression through alteration of these structures. In contrast, our studies show accumulation of 2,4-D in tubers of treated plants occurs and is associated with an enhanced tolerance to thaxtomin A. Applying 2,4-D to cultures of S. scabiei did not significantly alter in vitro growth of the pathogen. Thaxtomin A production by the pathogen was inhibited by 2,4-D, but only at the highest rate tested (1.0 mM), which is at least 200-fold more than is found in 2,4-D treated tubers. These data suggest 2,4-D has no direct effect on the pathogen or its virulence. Confirmatory evidence from studies with Arabidopsis thaliana seedlings demonstrated that the auxins 2,4-D and IAA ameliorate thaxtomin A toxicity. The evidence presented whereby auxin treatment inhibits toxicity of thaxtomin A secreted by the pathogen suggests a novel indirect means of disease suppression.

Occurrence of nutrients and plant hormones (cytokinins and IAA) in the water fern Salvinia molesta during growth and composting

The aquatic fern Salvinia molesta is classified as an invasive, noxious weed in southern Africa. It has rapid growth and forms a thick mat in slow moving water bodies. It is hypothesized that S. molesta is a suitable candidate to promote as an organic fertilizer. The aims of this study were to determine the mineral content as well as the cytokinin and auxin content in S. molesta. The release of these compounds into the culture medium and upon decomposition of the air-dried plant matter was also determined. Minerals detected in the fresh plant material were N > K > P > Ca > Mg > Fe > Na > Cu > Zn while Cd, Pb and Ni were below the level of detection. The mineral concentrations changed after 14 days decomposition with there being an increase in Ca, Mg and Fe and a decrease in K. Sixteen isoprenoid cytokinins were detected in the plant material (total concentration 81.62 pmol g −1 DW) while only four cytokinins occurred at a greatly reduced concentration in the culture medium (4.31 pmol 500 ml −1 medium). The material that had been composted for 14 days showed a 17% reduction in cytokinin concentration (67.96 pmol g −1 DW) compared to the plant material. Run-off, termed leachate, collected during the composting experiment also had high amounts of cytokinin-like activity measured using the soybean callus bioassay. This activity peaked on day 7 of the composting experiment, indicating a rapid release of cytokinin-like compounds. IAA occurred in much higher concentrations than the cytokinins in the plant material (393.6 pmol g −1 DW) while very little IAA was detected in the culture medium (30.8 pmol g −1 DW). IAA concentrations were much lower in the composted material (148.1 pmol g −1 DW) with a 62% loss being recorded compared to the plant material. No auxin-like activity was measured using the mungbean rooting bioassay in the leachate collected at the start of the composting experiment. However, the leachate collected on days 7 and 14 had biological activity indicating that auxin-like compounds were released from S. molesta upon decomposition.

Mutation ofE1-CONJUGATING ENZYME-RELATED1Decreases RELATED TO UBIQUITIN Conjugation and Alters Auxin Response and Development

The ubiquitin-like protein RELATED TO UBIQUITIN (RUB) is conjugated to CULLIN (CUL) proteins to modulate the activity of Skp1-Cullin-F-box (SCF) ubiquitylation complexes. RUB conjugation to specific target proteins is necessary for the development of many organisms, including Arabidopsis (Arabidopsis thaliana). Here, we report the isolation and characterization of e1-conjugating enzyme-related1-1 (ecr1-1), an Arabidopsis mutant compromised in RUB conjugation. The ecr1-1 mutation causes a missense change located two amino acid residues from the catalytic site cysteine, which normally functions to form a thioester bond with activated RUB. A higher ratio of unmodified CUL1 relative to CUL1-RUB is present in ecr1-1 compared to wild type, suggesting that the mutation reduces ECR1 function. The ecr1-1 mutant is resistant to the auxin-like compound indole-3-propionic acid, produces fewer lateral roots than wild type, displays reduced adult height, and stabilizes a reporter fusion protein that is degraded in response to auxin, suggesting reduced auxin signaling in the mutant. In addition, ecr1-1 hypocotyls fail to elongate normally when seedlings are grown in darkness, a phenotype shared with certain other RUB conjugation mutants that is not general to auxin-response mutants. The suite of ecr1-1 molecular and morphological phenotypes reflects roles for RUB conjugation in many aspects of plant growth and development. Certain ecr1-1 elongation defects are restored by treatment with the ethylene-response inhibitor silver nitrate, suggesting that the short ecr1-1 root and hypocotyl result from aberrant ethylene accumulation. Further, silver nitrate supplementation in combination with various auxins and auxin-like compounds reveals that members of this growth regulator family may differentially rely on ethylene signaling to inhibit root growth.