Effect of systemic fungicide (combination of Cyproconazole and propiconazole) newly introduced in Algeria on Septoria of two varieties of wheat (Triticum durum Desf).
Wheat can be attacked by many diseases at various stages of development. These attacks can cause significant losses when susceptible varieties are used and environmental conditions are favorable to the expansion of the disease.
Septoria of wheat caused by Mycosphaerella graminicola (Fuckel) Schroeter (anamorph Septoria tritici Rob. Ex Desm.) And Phaeosphaeria nodorum (E.Mull.) Hedjar. (anamorph Stagonospora nodorum (Berk.) E.Castell.Germano.) is a major wheat diseases worldwide . It can cause yield losses of up to 60% .
Septoria leaf spot disease is the most common wheat. The amount of rain upstream current is the first risk factor. This allows to mount sheet sheet [4,5].
During wet years, Septoria tritici is leading the parasitic complex of durum wheat in Algeria, losses can reach 50% .
The disease progresses until it reaches the first three leaves whose health status is important in grain filling, in order to establish a normal return.
Crop protection can be anticipated from the seedlings by the choice of varieties less sensitive and sometimes before sowing by affecting the choice of species in the rotation. We can therefore say that the fight against parasites is a fight that must multifactorial increasingly combine preventive measures, so-called prophylactic, with direct actions on the disease. Assessment of new active substances is still required to select and knowingly decide the best solutions the market offers pest. The effectiveness of the fight requires high-performance products and properly positioned in time. The susceptibility of varieties to leaf diseases is different, the struggle can be adapted accordingly . The continued increase in the use of treatment products demonstrates their remarkable ability to fit into the existing agricultural combinatorial techniques. So we see associated with the development of mechanization in field crops but also increasingly used in intensive culture in which they appear to have been primarily responsible for increases in productivity. Currently, 39% of these products are molecules and fungicides used to protect crops and crops against pests. However, their indiscriminate use can pose risks to consumers and dangerously destabilize ecosystems [8,9]. Even though most treatments are applied to the aerial parts of plants, much of the product always reaches the ground, where live bacteria, fungi, algae and insects. We should pay special attention to the harmful effects of pesticides on soil microflora, which is essential for maintaining fertility. Numerous studies have shown that treatments done correctly have limited effect on soil microbial metabolism .
Algeria is ranked among the countries that use larger amounts of pesticides in the world. And about 400 pesticides are registered in Algeria, including about forty varieties are widely used by farmers .
Our study fits into this theme, it evaluates the effectiveness of a systemic fungicide newly introduced in Algeria "ARTEA" on one of the major diseases of leaves and spikes affecting the yield of durum wheat: Septoria, caused by Septoria tritici conidial shape by analyzing the effects (physiological and biochemical tests) it causes on durum wheat in the study of changes in photosynthesis, metabolism and organic nutrients especially water potential.
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Materials and Methods
Our study focused on two varieties of wheat (Triticum durum): GTA hard and Bidi 17 having different geographical origins and with different tolerances to diseases as well. The seeds we are provided by The CETO Guelma.
The fungicide used is provided by the firm Syngenta is a systemic foliar fungicide with broad spectrum disease control for air as powdery mildew, septoria, rusts and tan spot on leaves of cereals , commonly called ARTEA 330 EC. It is presented as a light brown liquid, its active ingredient is composed of 80 g/l cyproconazole.et 250 g/l propiconazole
Presentation of the site of the experiment:
The experiment was conducted at the collective farm "Khrouf 1-1 Bachir" in El Kerma. It is situated in the plain of Annaba 10 km southeast of El-Hadjar Daira. Its latitude is 36[degrees]N and longitude 46mn Eastern is 7[degrees]38 minutes E, it is 28 Km from the sea experimental. Device
The experimental design adopted was a split plot with three replications and completely randomized. For each variety we have four basic plots of 2 m2 each: the first is treated with a concentration of 0.3 l/ ha of fungicide (D1) is treated with the second concentration of 0.4 l/ha of fungicide (D2), the third is treated with the concentration of 0.5 l/ha of the fungicide (D3), and represents the last. For each variety, each plot consists of six basic lines of 2 m long, spaced 20 cm, and the distance between two elementary plots is 1 m.
One treatment is considered curative applied at flowering stage of the two durum wheat varieties. Three concentrations are applied:
* D1 of 0.3 l/ha
* D2 of 0.4 l/ha
* D3 of 0.5 l/ha
The processing is done manually, using a spray bottle with a capacity of 1.2 l. Measured parameters:
To assess the degree of attack of the disease on two varieties of durum wheat used, seven readings or simultaneous ratings were made (15, 22, on April 29 and 06, 13, 20 and 27 May). To determine the intensity of the disease, a direct reading is done by determining the percentage of the surface of the sheet contested.
Physiological and biochemical analyzes:
Determination of water loss (RWL):
This is a cuticular transpiration evaluated by the method of  and determined using the following formula:
R.W.L = [[[W.sub.i] - [W.sub.2h]]/[W.sub.S]] 1/[[A.sub.1] x 120]
Wi = Initial weight of the sheet.
[W.sub.2h] = Weight of the sheet after 2 hours.
[W.sub.S] = Dry weight of the sheet.
[A.sub.1] = leaf area.
Determination of the water content (RWC):
It is measured with the method of Barrs (1968) in Ladigues (1975 ).) Using the following formula:
R.W.C = [[[W.sub.f] - [W.sub.s]]/[[W.sub.t] - [W.sub.s]]] X 100
Wf = Fresh weight of leaf excised.
Ws = Dry weight of the sheet.
Wt = weight of the sheet to turgor (weight after 24h in distilled water).
Energy parameter (measured as chlorophyll):
Chlorophyll content is determined by the traditional method of Mackiney (1941); in Holden . Measurement was made by the deduction of 3 ml of the solution into a cuvette spectrophotometer. Readings are taken at wavelengths of 645 and 663 nm, after calibration of the instrument with reference solution of 80% acetone. The total chlorophyll content is calculated taking into account the optical density for each type of chlorophyll a and b.
Chl. a = 12.7 (OD 663 nm) - 2.69 (OD 645 nm).
Chl. b = 22.9 (OD 645 nm) - 4.68 (OD 663 nm).
Chl. (a + b) = 8.02 (D.O. 663 nm) + 20.20 (D.O. 645 nm).
Determination of soluble sugars:
Soluble sugars were assayed following the method of Schields and Burnett . Using anthrone in sulfuric acid.
The optical density of the samples was determined using a spectrophotometer at wavelength 585 nm. For the correspondence between the optical density and concentration of soluble sugars, a standard range is produced using increasing concentrations of glucose, prepared from a stock solution of 100 g/ml.
Determination of proline:
The technique is that of Troll and Lindsley , Simplified and developed by Dreier and Goring , and then modified by Monneveux Nemmar .
It consists of extracting the proline of fresh plant material with alcohol (methanol) at 85[degrees]C for one hour and then to imprison in the ninhydrin reagent under acidic conditions (acetic acid) at 100[degrees]C for 30 minutes; a stable compound of red color is obtained, extracted with toluene. The optical density of the samples was determined using a spectrophotometer at wavelength 528 nm.
Correspondence between the optical densities and concentrations of proline is achieved through a standard range, obtained from increasing concentrations of proline, prepared from a stock solution of 0.2 mM.
The results are validated by a statistical study. Two tests were performed: the test of analysis of variance with one criterion of classification and testing of student.
Results and Interpretation:
Effect of ARTEA on the rate of infestation of two varieties of durum wheat (hard and BIDI17 GTA):
The results of treatment effects by ARTEA 330 Ec on the rate of infestation of plants of two varieties of durum wheat (hard and BIDI17 GTA) are shown in Figures 1 and 2
[FIGURE 1 OMITTED]
[FIGURE 2 OMITTED]
In Figure 1, we find infected plants (controls) that the variety is the first GTA hard to present the first symptoms of the disease, 90 days after emergence. It presents the infection rates highest with 10% which increases gradually to reach 39% (to 140 days after emergence) while for the variety Bidi 17, the infection rate began with 7% to the 28%.
In terms of the treaty, (the processing is performed 123 days after emergence), we note that it is always the hard variety GTA presenting infection rates highest with 8.5%, 13% and 15%. Whereas in the variety Bidi 17 (Figure 2), it is 7%, 11% and finally 12%.
Seven days after treatment, or 130 days after emergence, we note, in both (02) varieties and for the 3 concentrations, a total halt of infection.
Effect of ARTEA on the rate of water loss (RWL):
The results of the effect of ARTEA on the rate of water loss (RWL) of leaves of two varieties of durum wheat (hard and BIDI17 GTA) are presented in Figure 3 sont presentes dans la figure 3
[FIGURE 3 OMITTED]
The results obtained show that, for both (02) varieties, the fungicide has a positive effect on this parameter. In terms of witnesses, those of the hard variety GTA seem most affected. This first observation leads us to suggest that the latter is more sensitive than BIDI 17. Note that in plants treated in two varieties, those who received concentration (0.3 l /ha), show a RWL higher than those treated with concentrations (0.4 and 0.5 l/ha). Thus the fungicide reduced very significantly the RWL for both cultivars. It should be noted, among infected than among treated (with 03concentrations) is the variety GTA drive that shows the rate of water loss, the most important. Analysis of variance showed very highly significant effect between concentrations and between varieties, we note a reduction in the RWL highly significant (p < 0.001) D3 concentration sequentially for both varieties. While the Student test shows only a significant effect between the two varieties.
Effect of turgor on ARTEA on leaves (RWC):
The results for this parameter are presented in Figure 4.
[FIGURE 4 OMITTED]
The different results obtained show that the disease reduces the relative water content of leaves in two witnesses. Water retention at the leaves of wheat is affected. Indeed, we notice an increase in the RWC with the concentrations applied in ARTEA. For the different concentrations applied (D1, D2 and D3), the values of RWC highest are presented by the variety Bidi 17. This parameter confirms the results obtained with the RWL values that are lower in this same variety.
Statistical analyzes show very highly significant differences (P < 0 001) between the two varieties and between concentrations...
Effect of ARTEA on chlorophyll content of leaves and sugar:
Table 2 presents the results of these two parameters
Parameter for chlorophyll, the results in Table 2 show that this parameter is affected by the presence of pathogens in two varieties. The hard variety GTA is still the most affected by cons, the variety BIDI 17 has a higher chlorophyll content, and this, both in controls than in treated. The results of this parameter emphasize the positive effect exerted by the ARTEA on the synthesis of chlorophyll pigments or recorded higher levels for D2 and D3 levels.
All of these results suggests that the infection durum by pathogenic fungi is accompanied by a decrease in chlorophyll content, which reflects a decrease in the photosynthetic activity of leaves of control plants. These phenomena can be also explained by the decrease in leaf area or of its efficiency. Analysis of variance showed a highly significant effect between the different concentrations and both varieties, whereas the t test shows no significant effect at 5% between the two varieties.
[FIGURE 5 OMITTED]
In terms of total sugars (Tab 2), the results express very heterogeneous. And treated plants contain more sugars compared with control plants including concentrations of 0.4 l/ha and 0.5 l/ha for the variety and concentration BIDI17 0.3 l/ha and 0.4 l/ha for GTA drive. The lowest levels are those recorded in leaves of witnesses.
Effect of ARTEA on proline content of leaves: Figure 5 shows the results of this parameter La figure 5 presente les resultats de ce parametre
The results of this osmoticum showed a behavior very different varieties vis-a-vis the fungicide. The synthesis of proline appears to be stimulated by the presence of the fungicide. GTA on the hard variety, the highest rates are those quantified in the leaves treated with the concentration (0.3 l/ha). For this parameter is the variety GTA drive that reads this most important both in the treated than in infected (controls). The analysis of variance, shows a very highly significant effect between doses in both varieties. As for the Student test, it shows no significant effect between the two varieties.
The results obtained in our work show that the condition of the treated plants compared with control plants, resulting in a phenomenon of water loss (RWL) and a lower relative water content of leaves quite high.
In both cultivars, we note that the effect of the fungicide is very remarkable. The plants have an efficiency in treated water than in control plants (high water content, low attrition rate). This confirms the work of , showing that the action of different fungicides at different concentrations was reflected positively on the water status of leaves of durum wheat.
In general the variety GTA drive seems most affected by introducing the dropout rate highest, whereas the variety Bidi 17 seems to behave vis-a-vis the fungicide, it has a fairly high content of water from leaves and lower rate in RWL. This work confirms the observations of the research team of the CETO . Who report that the variety BIDI 17 is well known for its resistance to biotic agents. Thus the work of , show that the RWL and RWC are likely to increase or decrease as a result of infection and the test fluid is a response to infection. To , a stomatal resistance in wheat leaves infected by disease, is observed, it is linked on the one hand to an increase in the internal concentration of CO2 and on the other hand, a decrease of the conductance leaves and water loss through stomata. It is the same for leaf chlorophyll content, where the results indicate that this parameter is affected by the presence of pathogens in the 02 varieties studied And William Ayres ; Gethar et al ; Streuter et al., ; Magboul et al . And Robert and et al . reported that fungal pathogens are responsible for the wilting of leaves and a net decrease of photosynthesis, which causes premature senescence of wheat leaves infested.
For both varieties and at different concentrations of ARTEA, we notice a slight increase in chlorophyll content. The levels are slightly higher compared to controls. This seems in perfect agreement with the results obtained by Hammou, . And Bordjiba,  which show that fungicides have no significant effects on vital functions of the plant and in particular on the development of chlorophyll. It then used the pesticide as a source of carbon and energy . However, other authors such as Berova et al., ; Hennouni,  claim the opposite.
The values obtained show that the sugar content that osmoticum increases much more in the leaves of treated plants for both varieties compared to controls. This is explained by the fact that pathogens may use sugars as carbon source. This hypothesis is confirmed by the work of Minarcic and Janitor  in barley and wheat powdery mildew affected by these authors attribute this to the fact that during the early stages of infection, the content soluble sugars in infected plants, would be less than that of treated plants. Borjiba work , show that rates of carbohydrates are slightly higher in wheat plants treated, compared to control seedlings.
In terms of proline, this parameter is also an indicator of stress and this is confirmed by the values obtained in control plants of both varieties that have a proline content higher than that of treated. Several authors note changes in amino acid content and protein in tissues of plants infected with rust [34,35]. Hanson et al.,  reported that the rate of proline decreases rapidly in living tissues after oxidative stress or by incorporation of proline into proteins. This confirms the decrease and an increase in other. Proline as other compound, such as glutamine, glutathione and the phytol undergo changes in concentration and accumulate in the plant when the metabolic balance thereof is disturbed by the unfavorable environmental conditions (pollution, physiological stress, climatic factors). The concentration can vary from one plant to another and from one habitat to another [37,38]. The results of our study confirm this hypothesis, in fact, the variety GTA hard this content the most important both in the treated than in control compared to the variety BIDI17. accumulation of proline appears to be one of the most remarkable manifestations of the host-parasite interaction. Several authors have noted changes in the amino acid content and protein in tissues of plants infected with fungal diseases . Goring , attributed the accumulation of proline in plant tissues infected by pathogens with an increased rate of auxin. These results also reflect the prevalent disease-interaction with water stress resulted in alterations and severity coupled, thus justifying the high accumulation of this osmolyte.
This study allowed us to make an initial finding on the influence of a fungicide (330 ARTEA Ec at different concentrations (0.3 l/ha, 0.4 l/ha and 0.5 l /ha) and their impact on the physiology and biochemistry of two durum wheat varieties namely hard and GTA BIDI17. Results show that the fungicide tested greatly reduces recovery pycnidial and reductions in severity up to 80% compared with control plants. The rate of water loss of treated leaves drop significantly which leads to an increase in their relative water content. Chlorophyll and total carbohydrate were higher in treated plants compared with control plants. Finally, for proline, the treated plants, in general, have a content of this osmolyte, greater than that of control plants especially in the GTA Hard variety.
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(1) Hennouni, N., (2) Djebar, M.R. and (2) Djebar-Berrebbah, H.
(1) Agronomy Department, El Taref University Centre, El Taref, 36000, Algeria.
(2) Cell Toxicology Laboratory, Annaba University, Annaba, 23000, Algeria.
Hennouni, N., Djebar, M.R. and Djebar-Berrebbah, H: Effect of systemic fungicide (combination of Cyproconazole and propiconazole) newly introduced in Algeria on Septoria of two varieties of wheat (Triticum durum Desf)
Hennouni, N., Agronomy Department, El Taref University Centre, El Taref, 36000, Algeria.
Table 2: Chlorophyll content of leaves and sugar to the different treatments ARTEA in both varieties. d'ARTEA chez les deux varietes. parameters and Chlorophyll Sugar varieties [micro]g/g de M.F [micro]g/g de M.F GTA dur BIDI 17 GTA dur BIDI 17 Concentrations control 0,3 l/Ha 29,79 33,84 0,179 0,166 0,4 l/Ha 37,87 55,38 0,285 0,356 0,5 l/Ha 51,09 66,78 0,351 0,417 56,42 62,73 0,318 0,394