Naphthalic anhydride decreases persistence of alachlor and atrazine and elevates tolerance of maize.

The present study aimed at alleviating the impacts of alachlor (Ala) or atrazine (Atr) on maize growth by seed-dressing with naphthalic anhydride (NA, 0.4% w/w by seed weight). The dressed and undressed seeds were germinated for 10 days and treated with Ala or Atr at 3.2 or 1.8 kg ha-1, respectively, then the herbicide residues were determined in shoots one day after treatment. Atr residues were higher than Ala and remained in the same level for the 2nd day then retracted consistently up to 12 days. Meanwhile, GSH and GST were significantly induced while growth parameters were reduced; the effect of Atr was higher than Ala. Nonetheless, ABA, phenolics and anthocyanins as well as PAL, TAL and CI were increased but IAA was decreased coincidently with enhanced IAA-O and peroxidase. The immediate detection of the herbicide residues could conclude that growth reduction as elucidated from the decreased IAA concomitant with elevating ABA, phenolics and anthocyanin contents and enzyme activities are consequences of the herbicide persistence. The drop of IAA was preceded by the stimulation of IAA-O and peroxidase while the increased phenolics and anthocyanins followed PAL, TAL and CI stimulation confirming the regulatory roles of these enzymes. The application of NA greatly lowered the herbicide residues concurrently with ameliorations in growth parameters, GSH, GST, and maintained the balancing of secondary metabolites and plant growth regulators. Lowering Ala and Atr residues by NA in synchronization with enhanced GSH and GST could conclude that NA encouraged the detoxification of the herbicide. Moreover, the balances of IAA, ABA, phenolics and anthocyanins were mostly maintained in normal levels concomitantly with growth ameliorations suggesting that phytohormones and secondary metabolites are involved in the elevation of maize tolerance to Ala and Atr.

Kinetics of inhibition of isoproturon to glutathione-associated enzymes in wheat.

The present study aimed at investigating the kinetic of inhibition of isoproturon to the GSH-associated enzymes [γ-glutamyl-cysteine synthetase (γ-GCS), glutathione synthetase (GS), glutathione reductase (GR), glutathione-S-transferase (GST) and glutathione peroxidase (GPX)] in wheat. Isoproturon, applied to 10-day-old seedlings for the following 12 days, provoked significant reductions in shoot fresh and dry weights, protein, thiols and glutathione (GSH); however, oxidized glutathione (GSSG) was elevated while GSH/GSSG ratio was declined with concomitant significant inhibitions in the activities of γ-GCS, GS, GR, GST and GPX; the effect was time dependent. IC50 and Ki values of isoproturon were lowest for GPX, highest for both GST and GR, and moderate for both γ-GCS and GS. The herbicide markedly decreased Vmax of γ-GCS, GS and GPX but unchanged that of GST and GR; however, Km of γ-GCS, GS, GST and GR increased but unchanged for GPX. The pattern of response of changing Vmax, Km, Vmax/Km, kcat and kcat/Km for in vivo and in vitro tests of each enzyme seemed most likely similar. These results indicate that a malfunction to defense system was induced in wheat by isoproturon resulting in inhibitions in GSH-associated enzymes, the magnitude of inhibition was most pronounced in GPX followed by γ-GCS, GS, GST, and GR. These findings could conclude that isoproturon competitively inhibited GST and GR; however, the inhibition was noncompetitive for GPX but mixed for both γ-GCS and GS.

Exogenous application of spermine and putrescine mitigate adversities of drought stress in wheat by protecting membranes and chloroplast ultra-structure.

Polyamines (PAs) are positively charged molecules known to mitigate drought stress; however, little is known about their mechanism of alleviating drought stress. We investigated the effects of PAs exogenously applied as a seed primer and as a foliar spray on the growth, membrane stability (MS), electrolyte leakage (EL), Na+ and K+ cations, reactive oxygen species (ROS), catalase (CAT; EC 1.11.1.6) and guaiacol peroxidase (GPX; EC 1.11.1.7) activity and chloroplast ultra-structure in wheat (Triticum aestivum L.; cv. Sakha-94) under drought stress. Three PA solutions, namely, putrescine, spermine and a mixture of the two (Mix), were each applied at a concentration of 100 µM. Our study demonstrated that the retardation of chlorophyll loss and elevation of Rubisco levels were involved in PA-enhanced growth under drought stress. These relationships were mainly reflected in elevated fresh weight and dry weight in response to foliar spraying with all PA solutions and seed priming with the Mix solution. The elevated growth seemed to be due to increased photosynthetic pigments, protein and Rubisco. In contrast, drought decreased growth, photosynthetic pigments, protein and Rubisco. MS was enhanced by PAs applied as a seed primer or foliar spray, as shown by clear reductions in EL %, malondialdehyde (MDA) content and the Na+/K+ ratio as well as reduced ROS markers and elevated CAT (but not GPX) activity. Further study showed that the Mix solution of PAs, applied either during seed priming or as a foliar spray, improved chloroplast ultra-structure, suggesting that improvements in Rubisco and photosynthetic pigments were involved in PA maintenance of chloroplast stability. Therefore, the present study showed that elevated CAT activity is the main mechanism through which PAs reduce ROS and MDA, thereby improving MS and protecting mesophyll cells structurally and functionally under drought stress in wheat.

Exogenous applications of Polyamines modulate drought responses in wheat through osmolytes accumulation, increasing free polyamine levels and regulation of polyamine biosynthetic genes.

Polyamines (PAs) can improve drought stress tolerance in plants; however, very limited information is available on the mechanism of action of exogenous application by different methods under drought stress in wheat. The present study investigates the mechanism through which seed priming and foliar spraying with PAs protect wheat plants from drought stress. 10 days old wheat seedlings were exposed to drought stress by withholding water alone or with 100 µM PAs solutions (putrescine, Put; spermine, Spm; and mixture of Put and Spm for 10 h seed-priming or three foliar sprays during withholding water. Drought stress impaired the wheat growth and altered the osmoprotectants, endogenous PAs levels, PAs biosynthetic genes expression and weight of 1000 grains compared to the corresponding control values. Exogenously applied PAs improved cell water status, accumulated osmoprotectants and PAs and up-regulated PAs biosynthetic genes, ADC, arginine decarboxylase; DHS, deoxyhypusine synthase; ODC, ornithine decarboxylase and SAMDC, S-adenosyl methionine decarboxylase. Put significantly regulate the endogenous PAs by both methods of application, however, Spm and mixture of Put and Spm could positively regulate the endogenous PAs and the biosynthetic gene expression by foliar spraying rather than seed priming. The data provide evidence that maintenance of water economy through stabilized cellular structure is an important strategy of drought tolerance by PAs in wheat.

Efficacy of Caltropis procera and Ficus sycomorus extracts in treating MRSA (methicillin-resistant Staphylococcus aureus)-keratitis in rabbit.

MRSA-induced keratitis in rabbit was used to evaluate the therapeutic effect of F. sycomorus leaves and C. procera latex extracts. Within the 6 rabbit groups tested, group 1 received sterilized saline, while other groups (2 to 6) received 100 µl of intrastromal injections of 1.5×10(3) colony forming unit (cfu) ml(-1) of methicillin-resistant Staphylococcus aureus (MRSA). After 12 hours, groups 3 to 6 also received chloramphenicol, aqueous extract of C. procera latex, aqueous and alcoholic extracts of F. sycomorus leaves, respectively 3 times daily for 12 successive days. The tested extracts inhibited MRSA growth in vitro (i.e. on culture medium). Colony counts in cornea discs from groups 3 to 6 were significantly reduced (P ≤ 0.001) compared to group 2 (untreated). Clinical signs of keratitis were observed on group 2 until the end of experiment. In groups 3 to 6, gradual recovery was observed and signs disappeared by the 12(th) DPI (days post inoculation). Only mild symptoms persisted in group 5 (aqueous extract of leaves). In group 3 and 5, cornea, iris, ciliary body and conjunctiva showed mild leukocytic infiltration and depigmentation of melanin cells while recovery of cornea and iris was observed in groups 4 and 6. In conclusion, the used extracts have potential therapeutic effects on MRSA-induced keratitis in rabbit.

Roles of dehydrin genes in wheat tolerance to drought stress.

Physiological parameters and expression levels of drought related genes were analyzed in early vegetative stage of two bread wheat cultivars (Sids and Gmiza) differ in drought tolerance capacity. Both cultivars were imposed to gradual water depletion started on day 17 till day 32 after sowing. Sids, the more tolerant cultivar to drought showed higher fresh and dry weights than the drought sensitive genotype, Gmiza. Under water stress, Sids had higher membrane stability index (MSI), lower accumulated H2O2 and higher activity of the antioxidant enzymes; catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX) and superoxide dismutase (SOD) than Gmiza. On the other hand, the differential expression patterns of the genes dhn, wcor and dreb were observed due to water deficit intensity according to cultivar's tolerance to drought. The DNA sequence alignment of dun showed high similarity of about 80-92% identities with other related plants. The most striking overall observed trend was the highly induction in the expression of dun, wcor and dreb in leaves of the tolerant genotype, Sids under severe water stress.

Alleviation of isoproturon toxicity to wheat by exogenous application of glutathione.

Treatment with the recommended field dose of isoproturon to 7-d-old wheat seedlings significantly decreased shoot height, fresh and dry weights during the subsequent 15days. Meanwhile contents of carotenoids, chlorophylls and anthocyanin as well as activities of δ-aminolevulinate dehydratase (ALA-D), phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) were significantly inhibited. On the other hand, the herbicide significantly increased malondialdehyde (MDA), a naturally occurring product of lipid peroxidation and H2O2, while it significantly decreased the contents of glutathione (GSH) and ascorbic acid (AsA) and reduced the activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX). These findings indicate an induction of a stress status in wheat seedlings following isoproturon treatment. However, exogenous GSH appeared to limit the toxic effects of isoproturon and seemed to overcome this stress status. Most likely, contents of pigment and activities of enzymes were raised to approximate control levels. Moreover, antioxidants were elevated and the oxidative stress indices seemed to be alleviated by GSH application. These results indicate that exogenous GSH enhances enzymatic and nonenzymatic antioxidants to alleviate the effects of isoproturon.

Efficacy of a pyrimidine derivative to control spot disease on Solanum melongena caused by Alternaria alternata.

The pyrimidine derivative (4,6-dimethyl-N-phenyldiethyl pyrimidine, DPDP) was tested as a foliar spray fungicide at 50 mg l(-1) for protection of eggplant (Solanum melongena) from spot disease caused by Alternaria alternata. Varied concentrations of DPDP (10-50 mg l(-1)) differentially inhibited mycelial growth, conidial count and conidial germination of A. alternata growth in vitro; the magnitude of inhibition increased with increasing concentration. In vivo, an experiment was conducted in pots using a complete block randomized design and repeated twice with three replications and four treatments (control, A. alternata alone, DPDP alone and combination of DPDP and A. alternata) for 5 weeks (1 plant in pot × 3 pots per set (3 replications per treatment) × 4 sets (4 treatments) × 5 weeks × 2 experimental repetitions = 120 pots). In this experiment, 10-day-old eggplant seedlings were transplanted in pots and then inoculated with A. alternata, DPDP or their combination 1 week later. Leaves of the A. alternata-infected eggplant suffered from chlorosis, necrosis and brown spots during the subsequent 5 weeks. Disease intensity was obvious in infected leaves but withdrawn by DPDP. There were relationships between incidence and severity, greater in plant leaves infected A. alternata alone and diminished with the presence of DPDP. Moreover, the infection resulted in reductions in growth, decreases in contents of anthocyanins, chlorophylls, carotenoids and thiols as well as inhibitions in activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST). Nonetheless, the application of DPDP at 50 mg led to a recovery of the infected eggplant; the infection-induced deleterious effects were mostly reversed by DPDP. However, treatment with DPDP alone seemed with no significant impacts. Due to its safe use to host and the inhibition for the pathogen, DPDP could be suggested as an efficient fungicide for protection of eggplant to control A. alternata spot disease.

A possible role for C4 photosynthetic enzymes in tolerance of Zea mays to NaCl.

Treatment of 14-day-old maize cultivars (Hybrid351 and Giza2) with 250 mM NaCl significantly reduced shoot fresh and dry weights and protein content during the subsequent 12 days. The magnitude of reduction was more pronounced in Giza than Hybrid. Both cultivars contained converging levels of protein for the enzymes phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH), pyruvate phosphate dikinase (PPDK) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) under normal conditions; however, NaCl led to increase these levels in Hybrid and decrease them in Giza. Moreover, NaCl significantly inhibited the activities of PEPC, MDH and PPDK in both cultivars during the first 2 days, thereafter the inhibition nullified only in Hybrid; nonetheless, Rubisco was the least affected enzyme in both cultivars. In addition, NaCl slightly increased V (max) of PEPC, MDH and PPDK in Hybrid with no change in K (m); nevertheless V (max) dropped in Giza with an increase in K (m) of only PEPC and MDH. Also K (cat), K (cat)/K (m) and V (max)/K (m) of all enzymes were lower in treated Giza than in treated Hybrid. The increased V (max) of all enzymes in only Hybrid by NaCl confirms that they were synthesised more in Hybrid than in Giza. However, the decreased V (max) in Giza concomitant with the increased K (m) points to an interference of salinity with synthesis of enzymes and their structural integrity. This would lead to a noncompetitive inhibition for the enzymes. These findings declare that maize tolerance to NaCl was larger in Hybrid compared to Giza due to a role for C4 enzymes.

Granular formulation of Fusarium oxysporum for biological control of faba bean and tomato Orobanche.

BACKGROUND: Orobanche spp. represent a serious threat to a wide range of crops. They are difficult targets for herbicides, and biological control could provide a possible solution. This work therefore aimed to formulate mycoherbicides of Fusarium with adequate shelf life and virulence against Orobanche but safe to faba bean and tomato. RESULTS: Only two isolates of Fusarium oxysporum Schlecht. (Foxy I and Foxy II) obtained from diseased Orobanche shoots were found to be pathogenic to Orobanche crenata Forsk. and Orobanche ramosa L. Conidial suspension of both isolates significantly decreased germination, attachments and tubercles of Orobanche. Microconidia and chlamydospores of both isolates were formulated as mycoherbicides encapsulated in a wheat flour-kaolin matrix (four different formulations). All formulations greatly diminished Orobanche emerged shoots, total shoot number, shoot height, attachment of emerged shoots, the germinated seeds that succeeded in emerging above the soil surface and dry weight. Meanwhile, disease incidence and disease severity of emerged shoots were enhanced. The shelf life was adequate, particularly for coarse, freshly prepared, low-temperature-stored, microconidia-rich formulations. The induced growth reduction of Orobanche-infected host plants seemed to be nullified by formulations, particularly at the highest dose. CONCLUSION: These formulations seemed to destroy Orobanche but appeared harmless to host plants. Hence, they could be efficiently used as mycoherbicides for biological control of Orobanche in faba bean and tomato.