Joseph Hershenhorn

Strigolactones affect lateral root formation and root-hair elongation in Arabidopsis

Strigolactones (SLs) have been proposed as a new group of plant hormones, inhibiting shoot branching, and as signaling molecules for plant interactions. Here, we present evidence for effects of SLs on root development. The analysis of mutants flawed in SLs synthesis or signaling suggested that the absence of SLs enhances lateral root formation. In accordance, roots grown in the presence of GR24, a synthetic bioactive SL, showed reduced number of lateral roots in WT and in max3-11 and max4-1 mutants, deficient in SL synthesis. The GR24-induced reduction in lateral roots was not apparent in the SL signaling mutant max2-1. Moreover, GR24 led to increased root-hair length in WT and in max3-11 and max4-1 mutants, but not in max2-1. SLs effect on lateral root formation and root-hair elongation may suggest a role for SLs in the regulation of root development; perhaps, as a response to growth conditions.

Strigolactones interact with ethylene and auxin in regulating root-hair elongation in Arabidopsis

Strigolactones (SLs) or derivatives thereof have been identified as phytohormones, and shown to act as long-distance shoot-branching inhibitors. In Arabidopsis roots, SLs have been suggested to have a positive effect on root-hair (RH) elongation, mediated via the MAX2 F-box. Two other phytohormones, auxin and ethylene, have been shown to have positive effects on RH elongation. Hence, in the present work, Arabidopsis RH elongation was used as a bioassay to determine epistatic relations between SLs, auxin, and ethylene. Analysis of the effect of hormonal treatments on RH elongation in the wild type and hormone-signalling mutants suggested that SLs and ethylene regulate RH elongation via a common regulatory pathway, in which ethylene is epistatic to SLs, whereas the effect of SLs on RH elongation requires ethylene synthesis. SL signalling was not needed for the auxin response, whereas auxin signalling was not necessary, but enhanced RH response to SLs, suggesting that the SL and auxin hormonal pathways converge for regulation of RH elongation. The ethylene pathway requirement for the RH response to SLs suggests that ethylene forms a cross-talk junction between the SL and auxin pathways.

Strigolactones’ Effect on Root Growth and Root-Hair Elongation May Be Mediated by Auxin-Efflux Carriers

Strigolactones are a new group of plant hormones that play a pivotal role in the regulation of aboveground plant architecture. However, the mechanisms governing their regulation of plant growth and development are unknown. We characterized the effect of a synthetic strigolactone (GR24) on tomato (Solanum lycopersicon) roots and present evidence for its relationship with the plant hormone auxin. We demonstrate that strigolactones interfere with the inhibitory effect of exogenously applied auxin on root elongation. This GR24-induced root elongation is conveyed via an increase in root cell length accompanied by a reduction in cell diameter, and it occurs despite strigolactone’s reduction of cell division (detected as reduction of CYCB1;1 transcript). In addition, high concentrations of strigolactone lead to asymmetric root growth and inhibition of root-hair elongation. Exogenous application of NAA or IAA was unable to restore symmetric root growth and root-hair elongation in the presence of strigolactone. However, application of NPA, an auxin-efflux inhibitor, did restore root-hair elongation in the presence of strigolactone. Similarly, exogenous application of 2,4-D, a synthetic auxin that is not secreted by efflux carriers, restored root-hair elongation and symmetric growth in the presence of strigolactone. Nevertheless, 2,4-D was unable to prevent root elongation by strigolactones. Therefore, strigolactones’ effect on root growth and root-hair elongation appears to be mediated via an effect on auxin-efflux carriers. Nevertheless, more than one mechanism may govern strigolactones’ effect on root growth.

A tomato strigolactone-impaired mutant displays aberrant shoot morphology and plant interactions

Strigolactones are considered a new group of plant hormones. Their role as modulators of plant growth and signalling molecules for plant interactions first became evident in Arabidopsis, pea, and rice mutants that were flawed in strigolactone production, release, or perception. The first evidence in tomato (Solanum lycopersicon) of strigolactone deficiency is presented here. Sl-ORT1, previously identified as resistant to the parasitic plant Orobanche, had lower levels of arbuscular mycorrhizal fungus (Glomus intraradices) colonization, possibly as a result of its reduced ability to induce mycorrhizal hyphal branching. Biochemical analysis of mutant root extracts suggested that it produces only minute amounts of two of the tomato strigolactones: solanacol and didehydro-orobanchol. Accordingly, the transcription level of a key enzyme (CCD7) putatively involved in strigolactone synthesis in tomato was reduced in Sl-ORT1 compared with the wild type (WT). Sl-ORT1 shoots exhibited increased lateral shoot branching, whereas exogenous application of the synthetic strigolactone GR24 to the mutant restored the WT phenotype by reducing the number of lateral branches. Reduced lateral shoot branching was also evident in grafted plants which included a WT interstock, which was grafted between the mutant rootstock and the scion. In roots of these grafted plants, the CCD7 transcription level was not significantly induced, nor was mycorrhizal sensitivity restored. Hence, WT-interstock grafting, which restores mutant shoot morphology to WT, does not restore mutant root properties to WT. Characterization of the first tomato strigolactone-deficient mutant supports the putative general role of strigolactones as messengers of suppression of lateral shoot branching in a diversity of plant species.

The synthetic strigolactone GR24 influences the growth pattern of phytopathogenic fungi

Strigolactones that are released by plant roots to the rhizosphere are involved in both plant symbiosis with arbuscular mycorrhizal fungi and in plant infection by root parasitic plants. In this paper, we describe the response of various phytopathogenic fungi to the synthetic strigolactone GR24. When GR24 was embedded in the growth medium, it inhibited the growth of the root pathogens Fusarium oxysporum f. sp. melonis, Fusarium solani f. sp. mango, Sclerotinia sclerotiorum and Macrophomina phaseolina, and of the foliar pathogens Alternaria alternata, Colletotrichum acutatum and Botrytis cinerea. In the presence of this synthetic strigolactone, intense branching activity was exhibited by S. sclerotiorum, C. acutatum and F. oxysporum f. sp. melonis. Slightly increased hyphal branching was observed for A. alternata, F. solani f. sp. mango and B. cinerea, whereas suppression of hyphal branching by GR24 was observed in M. phaseolina. These results suggest that strigolactones not only affect mycorrhizal fungi and parasitic plants, but they also have a more general effect on phytopathogenic fungi.

Indole-3-acetic acid biosynthetic pathways in Erwinia herbicola in relation to pathogenicity on Gypsophila paniculata☆

Pathogenic strains of Erwinia herbicola incite crown and root galls in the flowering ornamental gypsophila. Both pathogenic and non-pathogenic strains of the bacterium readily produce indole-3-acetic acid in culture. Two pathways for biosynthesis of indole-3-acetic acid were identified in E. herbicola: (1) the indole-3-acetamide route occurs via the following reactions: l-tryptophan → indole-3-acetamide → indole-3-acetic acid, and (2) the indole-3-pyruvate route involves the following reactions: l-tryptophan → indole-3-pyruvate → indole-3-acetaldehyde → indole-3-acetic acid. Production of indole-3-ethanol was also linked to the latter pathway. Evidence for the existence of the two pathways was based on: (a) chemical identification of the respective indole intermediates by thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectroscopy; (b) production of indole-3-acetic acid by bacterial cells treated with the various indole intermediates; and (c) incorporation of 3-14C-l-tryptophan into the indole intermediates of the two pathways. In contrast to the indole-3-pyruvate pathway which was detected in all the pathogenic and non-pathogenic strains examined, the indole-3-acetamide pathway was detected only in the pathogenic strains of E. herbicola. The possible relationship between the indole-3-acetamide pathway and gall formation by E. herbicola is discussed.

Broomrape (Orobanche cumana) Control in Sunflower (Helianthus annuus) with Imazapic1

Abstract: Field trials were conducted in 1997 and 1998 at two locations in Israel to evaluate the efficacy of imazapic applied postemergence (POST) to sunflower for broomrape control under irrigated and nonirrigated conditions. Two sequential treatments of imazapic at 1.5 followed by (FB) 3.0, 3.0 FB 4.5, or 4.5 FB 6.0 g ai/ha on sunflower plants 12 ± 3 and 55 ± 5 cm tall, respectively, reduced sunflower broomrape throughout the growing season under irrigated and nonirrigated conditions. Sunflower growth was not affected by imazapic treatments. It was confirmed, in accordance with an earlier report, that when sequential treatments of imazapic included an application at the sunflower inflorescence developmental stage, the herbicide decreased seed yield in proportion to the applied rate. Nomenclature: Imazapic; broomrape, Orobanche cumana Waller; sunflower, Helianthus annuus L. Additional index words: Parasitic weed, herbicide. Abbreviations: fb, followed by; POST, postemergence; PRE, preemergence.

Characterization of a novel tomato mutant resistant to the weedy parasites Orobanche and Phelipanche spp.

Orobanche and Phelipanche, commonly known as broomrape, are dicotyledonous holoparasitic flowering plants that cause heavy economic losses in a wide variety of plant species. Breeding for Orobanche resistance is still one of the most effective management strategies for this weed. However, previous efforts to find broomrape-resistant tomato (Solanum lycopersicon) genotypes have been unsuccessful. Here, we report on the isolation and characterization of a fast-neutron-mutagenized M-82 tomato mutant, Sl-ORT1. The Sl-ORT1 mutant showed resistance to Phelipanche aegyptiaca as compared to cultivar M-82; segregation analysis suggested a single recessive ort1 allele. Sl-ORT1 broomrape resistance was reflected in a lower number of broomrapes per plant, reduced P. aegyptiaca fresh weight per plant, and the absence of broomrape’s negative effect on plant host growth and yield. Sl-ORT1 was shown to be resistant to high concentrations of P. aegyptiaca seeds, and to another three broomrape species: Phelipanche ramosa, Orobanche cernua, and Orobanche crenata. Grafting experiments demonstrated that roots, rather than shoots, are necessary for Sl-ORT1 broomrape resistance. In addition, Sl-ORT1 was shown to be resistant to broomrape under field conditions. Since yield parameters are slightly affected by the mutation, this resistance gene should be introduced into tomato varieties with different genetic backgrounds; this newly identified Orobanche-resistant mutant may be further utilized in breeding programs for Orobanche resistance.

Strigolactone Deficiency Confers Resistance in Tomato Line SL-ORT1 to the Parasitic Weeds Phelipanche and Orobanche spp.

The parasitic flowering plants of the genera Orobanche and Phelipanche (broomrape species) are obligatory chlorophyll-lacking root-parasitic weeds that infect dicotyledonous plants and cause heavy economic losses in a wide variety of plant species in warm-temperate and subtropical regions. One of the most effective strategies for broomrape control is crop breeding for broomrape resistance. Previous efforts to find natural broomrape-resistant tomato (Solanum lycopersicon) genotypes were unsuccessful, and no broomrape resistance was found in any wild tomato species. Recently, however, the fast-neutron-mutagenized tomato mutant SL-ORT1 was found to be highly resistant to various Phelipanche and Orobanche spp. Nevertheless, SL-ORT1 plants were parasitized by Phelipanche aegyptiaca if grown in pots together with the susceptible tomato cv. M-82. In the present study, no toxic activity or inhibition of Phelipanche seed germination could be detected in the SL-ORT1 root extracts. SL-ORT1 roots did not induce Phelipanche seed germination in pots but they were parasitized, at the same level as M-82, after application of the synthetic germination stimulant GR24 to the rhizosphere. Whereas liquid chromatography coupled to tandem mass spectrometry analysis of root exudates of M-82 revealed the presence of the strigolactones orobanchol, solanacol, and didehydro-orobanchol isomer, these compounds were not found in the exudates of SL-ORT1. It can be concluded that SL-ORT1 resistance results from its inability to produce and secrete natural germination stimulants to the rhizosphere.

Egyptian Broomrape (Orobanche aegyptiaca) Control in Tomato with Sulfonylurea Herbicides—Greenhouse Studies1

Broomrapes (Orobanche spp.) are root holoparasitic plants that cause severe damage to economically important crops, especially in Mediterranean countries. Egyptian broomrape is the most troublesome weed on tomatoes grown for processing in Israel. In the present study, we tested the efficacy and selectivity of four sulfonylurea herbicides in controlling Egyptian broomrape on tomatoes grown in pots under greenhouse conditions. MON 37500, rimsulfuron, HOE 404 and SL-160 were applied postemergence (POST) and preplant incorporated (PPI) followed by POST applications. MON 37500 and rimsulfuron were more selective to tomato and controlled the parasite more effectively than HOE 404 and SL-160. MON 37500 and rimsulfuron at 50 and 100 g ai/ha and at 100, 150, and 200 g ai/ha, respectively, applied on tomato foliage 14, 28, and 42 d after planting (DAP) and followed by sprinkler irrigation to field capacity, resulted in complete control of the parasite. However, a significant reduction in control efficacy was observed when the experiment was repeated with charcoal-topped pots, suggesting that the herbicides act mainly through the soil. Except for rimsulfuron, the PPI followed by two POST treatments was more phytotoxic to tomato plants than the POST treatments. The PPI plus POST applications controlled Egyptian broomrape effectively, but tomato plants were injured by HOE 404 at all PPI application rates and by MON 37500 at the high rate at 150 g/ha. The present study determined that three POST applications or a PPI application followed by two POST applications of MON 37500 at 50 or 100 g/ha, or rimsulfuron at 100, 150, or 200 g/ha were effective and selective in controlling Egyptian broomrape on tomato, under greenhouse conditions. Nomenclature: HOE 404, 2-ethoxyphenyl [[(4,6-dimethoxy-2-pyrimidinyl) amino]carbonyl]sulfamate; MON 37500, N-[[(4,6-dimethoxy-2-pyrimidinyl) amino] carbonyl]-2-(ethylsulfonyl)imidazo[1,2-a]pyridine-3-sulfonamide; rimsulfuron; SL-160, N-[[(4,6-dimethoxy-2-pyrimidinyl)amino] carbonyl]-3-(trifluoromethyl)-2-pyridinesulfonamide; Egyptian broomrape, Orobanche aegyptiaca Pers. #3 ORAAE; tomato, Lycopersicon esculentum Mill. Additional index words: Activated charcoal, Orobanche control, parasitic weeds. Abbreviations: DAP, days after planting; POST, postemergence; PPI, preplant incorporated.