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Research Article

volume 41 issue 3 (september 2021) : 450-454,   Doi: 10.18805/ag.D-304

Biocontrol of Bayoud Disease (Fusarium oxysporum F. Sp. Albedinis) on Deglet-nour Variety of Date Palm (Phoenix dactylifera L.) in South Western Oases of Algeria

H
Hakima Belaidi1,*
F
Fawzia Toumi-Benali1
I
Ibrahim Elkhalil Benzohra2
1Laboratoire Ecodeveloppement des Espaces, Department of Environmental Sciences, Djillali Liabes University of Sidi Bel Abbes, Algeria.
2Department of Phoeniciculture, Experimental Station for Biophysical Environment Bechar (Taghit)-Centre for Scientific and Technical Research on Arid Regions (CRSTRA), Campus Universitaire B.P. 1682 RP, Biskra, Algeria.
Cite article:- Belaidi Hakima, Toumi-Benali Fawzia, Benzohra Elkhalil Ibrahim (2021). Biocontrol of Bayoud Disease (Fusarium oxysporum F. Sp. Albedinis) on Deglet-nour Variety of Date Palm (Phoenix dactylifera L.) in South Western Oases of Algeria . Agricultural Science Digest. 41(3): 450-454. doi: 10.18805/ag.D-304.

ABSTRACT

Background: This work has the objective to biocontrol the bayoud disease of date palm caused by the fungus Fusarium oxysporum f. sp. albedinis (Foa), is the major disease in Algerian palm groves of south western region. This in vivo biocontrol was done on date palm variety Deglet-Nour, using two antagonistic fungi species, Trichoderma harzianum and Aspergillus flavus.

Methods: Twenty Foa isolates obtained from isolation of the spines carrying the typical symptoms of Bayoud disease were used. Two strains T. harzianum and A. flavus, were isolated from rhizosphere soils of the date palm trees. Using the greenhouse screening test, which was carried out on 3-4 month-old date palm seedlings at the rate of 5 ml of inoculum suspension.

Result: A statistical analysis showed a significant (P<0.05), difference of Deglet-Nour seedlings reactions against Foa isolates and antagonistic fungi were observed. All date palm trees present susceptibility against Foa until the total mortality of seedlings. The biocontrol test showed that two antagonists showed different reaction, with the complete resistance for seedlings treated by T. harzianum with the mortality rate (rm%) reduced at 100%, while, the second antagonist A. flavus reduced the rm% at 25%. We can apply of these antagonistic fungi to protect our groves contaminated by Bayoud disease and also contain this susceptible commercial variety.

INTRODUCTION

The date palm (Phoenix dactylifera L.) is considered a symbol of life in the desert, while it tolerates high temperatures, drought and salinity more than other crop plant species. It is one of the oldest trees from which man has derived benefit and it has been cultivated since ancient times (Zohary and Hopf, 2000). It the only indigenous wild desert plant definitely domesticated in its native harsh environments appears to be the date palm (Zohary and Hopf, 2000 ; Vinita and Punia, 2016). Among the most cultivated and important varieties in Algeria, Deglet Nour, considered as the golden fruit of the oases, means: Deglet: light; Nour: light, is the best variety in the world (Munier, 1974). The Algerian palm groves represent 45% of Deglet Nour variety cultivation (Bouguedoura et al., 2008). The annual production of Deglet-Nour variety of dates is 5670000 quintals (Statistica, 2020), which represents 51,8% of total Algerian date palm production (FAO, 2020). The date palm sector has many problems like sand dunes, salinity, secheresse and seriously damaged by Bayoud disease (Touzi, 2007; Benzohra et al., 2015), which destroyed a half date palm trees of Algerian palm groves in the south-west region (Bouguedoura et al., 2008).


Bayoud disease caused by the soil fungus Fusarium oxysporum f. sp. albedinis (Foa), is considered as the most destructive fungal disease in south west of Algerian palm groves (Djerbi, 2003). It’s originated from Morocco and its progression continues to advance from the west and central of Algerian palm groves (Benzohra et al., 2015; 2017). The proposal used of prophylactic measures against this disease doesn’t seem able to stop progressive spread of this disease (Benzohra et al., 2017). Other method was used, a genetic control by resistant varieties is therefore another promising way to reduce this constraint, but unfortunately all resistant cultivars haven’t high food quality in date fruits (Sedra, 2005 a,b). It is necessary to protect this variety (Deglet-Nour), against this constraint with an effective method that does not pollute the environment. The biological control remains the best way to combat this constraint to protect this variety (El-Hassni et al., 2007; Jaiti et al., 2006, Jaiti et al., 2008; Kala et al., 2016; Fitrianingsih et al., 2019).         


In this case, this study has the objective to evaluate the biocontrol capacity of two antagonistic fungi species, Trichoderma harzianum and Aspergills flavus on the incidence of bayoud disease by testing of the important commercial date palm variety, Deglet Nour, but susceptible to Bayoud diseases.

MATERIALS AND METHODS

Date palm variety’s seedlings
 
The date palm variety used in this test named Deglet-Nour, this Algerian variety, has the high quality of date fruit in the word (Munier, 1974).
 
Fungal material
 
Twenty isolates of Foa were used in the inoculation test of date palm seedlings, was sampled from contaminated palm groves in Saoura, Gourrara, Touat and M’zab provinces in 2016 (Table 1). The antagonistic microorganisms were obtained from soils of date palm rhizosphere of Bechar region in 2019 (Table 2). These antagonistic fungi used in this study provided by CRSTRA institute (Center for Scientific and Technical Research on Arid Regions, Biskra, Algeria), from Laboratory of Plant Pathology and Protection, of Experimental Station of Biophysical Environment, Naama, Algeria. The isolates were conserved in PDA cultures (For each 1000 ml of distillated water: 200 g potato filtrate; 15 g glucose; 20 g agar-agar; pH=5,5), with temperature of conservation at 04°C and incubation at 23°C (El-Hassni et al., 2007).
 

Table 1: Foa isolates with its origin.


 

Table 2: Informations of antgonistic fungi strains’ species used in ‘in vivo’ biocontrol test against bayoud disease.


 
Preparation of inoculum
 
Date palm seedlings of Deglet-Nour variety of 2-3 months-old, were obtained by pre-germination of date palm seeds (Fig 1), after disinfection with Hypochlorite of Sodium at 2%, then rinsed with sterile distilled water three successive times (Sedra, 2005b). The date palm seeds were placed in Petri dishes containing the cotton soaked with sterile distilled water and then stored in the oven at a temperature of 23±2°C (El-Hassni et al., 2007). After 20 days of pre-germination (Fig 1), the seeds were transplanted into pots containing sterile peat for two months (Sedra, 1985).
 

Fig 1: Pre-germinatings seeds of Deglet-Nour variety used for test of biocontrol.


 
The in vivo biocontrol test was performed using the mixture ‘Foa inoculum + antagonistic fungus’ and two controls, one for seedlings treated with Foa inoculum alone and other without Foa inoculum. Four seedlings were used per treatment and repetition. After inoculation tests, the seedlings were conserved in small greenhouse at 25±2°C with 16 h of photoperiod.
 
Rating scale
 
The mortality rate of date palm seedlings was based on rate of mortality and calculations of dead plants as compared to all the plants inoculated (Sedra, 2005b). The mortality rate (mr) formula was as follows:

Statistical analysis
 
The experiment design was randomized complete blocks. Four replicates (number of seedlings tested), per treatment were taken and each whole experiment was performed independently. All collected data were submitted to ANOVA analysis using XLStat software 9 (AddinSoft, USA Ltd.) and the significance of differences was recorded at P<0.05, by Newman-Keuls test.

RESULTS AND DISCUSSION

The results of statistical analysis showed significant differences (P0.05), between the date palm varieties and also between two antagonistic microorganisms (Table 3). The results showed a high level of reduced mortality rate (mr) due to two antagonists of incidence of Bayoud disease on deglet-Nour variety seedlings compared to the control.   The control (direct inoculation with Foa alone), presented a very high mortality rate up to 100% (Table 3), while the control without Foa isolates inoculums had no typical symptoms of bayoud (Fig 2). Absences of symptoms were observed on seedlings inoculated by the antagonistic fungus T. harzianum with no mr (%). The second antagonistic fungus A. flavus could reduced the mr at 25% of seedlings tested (Table 3). In comparison between these two antagonistic species, we confirm that T. harzianum was more efficient than A. flavus.
 

Fig 2: Reactions of Deglet-Nour variety’s seedling against Bayoud disease in presence of two antagonistic fungi species. Foa: Fusarium oxysporum f. sp. albedinis; antagonistic fungi species: T. harzianum: Trichoderma harzianum, A.flavus.: Aspergillus flavus; *: date palm varieties names.


 

Table 3: Mortality rates and reaction of Deglet-Nour variety seedlings of date palm in the biocontrol test against bayoud disease.


 
Our study showed that the antagonistic fungal species tested have significant biocontrol activity against Bayoud disease when tested in vivo. The mortality rate (mr) varied significantly ascompared tocontrol, when compared with two antagonistic fungi, T. harzianum was more efficient with mr=0%, than A. flavus with mr =25%.
       
Many reports have been published about the importance of biocontrol as the best way to combat Fusarium wilt (Sajeena et al., 2020; Cheng et al., 2019; Ou-Zine et al., 2020). The importance of antagonistic fungi like T. harzianum as biofungicide against Fusarium wilt of many crops has been reported by Kala et al., (2016), including Fusarium wilt of chickpea (Cicer arietinum L.) caused by Fusarium oxysporum f. sp. ciceris, by Sahampoor et al., (2020), on Fusarium wilt of cucurbits caused by Fusarium oxysporum f. sp. radicis-cucumerinum and also by Fitrianingsih et al., (2019), on Fusarium wilt of tomato (Lycopersicum esculentum L.) caused by the soil fungus Fusarium oxysporum f. sp. radicis-lycopersici.
 
The antagonistic fungus Aspergillus flavus was also reported by many authors effective against Fusarium wilt diseases. Among these, Fusarium oxysporum f. sp. melonis of melon (Cucumis melo L.), Fusarium oxysporum f. sp. vasinfectum of Cotton (Gossypium herbaceum L.), (Dong and Cohen, 2002), Fusarium graminearum of cereals (Mejdoub-Trabelsi et al., 2017) and Rhizoctonia solani (Vibha, 2010) and Fusarium sambucinum of potato (Solanum tuberosum L.), (Aydi et al., 2015).
 
It can therefore be concluded that two antagonistic fungi species (T. harzianum and A. flavus), can enhance the ability to of host to resist bayoud disease in Deglet-Nour variety under in vivo biocontrol tests in palm groves in Algeria.

CONCLUSION

This study had the objective to evaluate the capacity of in vivo biocontrol on incidence and aggressiveness of Bayoud disease caused by Fusarium oxysporum f. sp. albedinis (Foa), which is  the major constraint of date palm cultivation in south-western palm groves of Algeria. This biocontrol was realized by use of two antagonistic fungal species Trichoderma harzianum and Aspergillus flavus, against the important variety of date palm in Algeria named Deglet-Nour. The results showed significant differences between two antagonistic fungi as well as the control (inoculation with and without Foa strains mixture inoculum). In comparison between these two antagonistic species, we can say that T. harzianum is more efficient than that species A. flavus, with mr % values  of 0%, 25%, respectively.
 
The use of biological agents manage disease in the susceptible varieties like Dglet-Nour against Bayound provide an ecologically friendly alternative to pesticides to control the disease. This strategy has more significance against soil-borne pathogens such as Fusarium wilt in palm groves of south-west region of Algeria.

ACKNOWLEDGEMENT

We would like to thanks the DGRSDT (Direction Générale de la Recherche Scientifique et du Développement Technologique, Algiers, Algeria), affiliated with the Ministry of High Education and Scientific Research of Algeria, for its funding support which facilitated us to realize this work in  the  Social – Economical project. 

REFERENCES


  1. Aydi, B.A.R., Jabnoun-Khiareddine, H., Mejdoub-Trabelsi, B., Daami-Remadi, M. (2015). Soil-borne and Compost-borne Aspergillus Species for Biologically Controlling Post-harvest Diseases of Potatoes Incited by Fusarium sambucinum and Phytophthora erythroseptica. Journal of Plant Pathology and Microbiology. 06: 313. doi: 10. 4172/2157-7471.1000313.

  2. Benzohra, I.E., Megateli, M., Berdja, R. (2015). Bayoud disease of date palm: History, epidemiology and integrated disease management. African Journal of Biotechnology. 14(7): 542-550. Doi 10.5897/AJBX2014.14292.

  3. Benzohra, I.E., Megateli, M., Elayachi, B.A., Zekraoui, M., Djillali, K., Bouafia, A., Benouis, S., Benaziza, A., Rekis, A. (2017). Integrated management of Bayoud disease on date palm (Phoenix dactylifera L.) caused by Fusarium oxysporum f. sp. albedinis in Algeria. Journal Algérien des Régions Arides. 14: 93-100.

  4. Bouguedoura, N., Ibrahim, A., Ould Mohamed, A., Saker, M., Trifi, M. (2008). A paper presented at NEPAD Biotechnology Workshop “Challenges for North Africa and Promises for a Regional Integrated Program”, Ezzahra, Tunisia, in collaboration with the Tunisian Society of Microbiology. pp. 22-25.

  5. Cheng, C., Li, C., Qi, Q., Sun, X., Anue, M.R., Mahoudjro David, B., Zhang, Y., Hao, X., Zhang, Z., Lai, Z. (2019). The root endophytic fungus Serendipita indica improves resistance of Banana to Fusarium oxysporum f. sp. cubense tropical race 4. European Journal of Plant Pathology. 156: 87-100. doi 10.1007/s10658-019-01863-3

  6. Djerbi, M. (2003). Fusarium oxysporum f. sp. albedini. OEPP/EPPO Bulletin. 33: 245-247.

  7. Dong, H., Cohen, Y. (2002). Induced resistance in cotton seedlings against Fusarium wilt by dried biomass of Penicillium chrysogenum and its water extract. Phytoparasitica. 30: 77-87.

  8. El-Hassni, M., El-Hadrami, A., Daayf, F., Cherif, M., Ait Barka, E., El-Hadrami, I. (2007). Biological control of bayoud disease in date palm: Selection of microorganisms inhibiting the causal agent and inducing defense reactions. Environmental and Experimental Botany. 59: 224-234.

  9. FAO. (2020). FAO Statistical Yearbook suite of products. http://www.fao.org/faostat/en/#data/QC

  10. Fitrianingsih, A., Martanto, E.A., Barahima, A. (2019). The effectiveness of fungi Gliocladium fimbriatum and Trichoderma viride to control fusarium wilt disease of tomatoes (Lycopersicum esculentum). Indian Journal of Agricultural Research. 53(1): 57-61. doi 0.18805/IJARe.A-363

  11. Jaiti, F., Meddich, A., El-Hadrami, I. (2006). Effect of arbuscular mycorrhization on the accumulation of hydroxycinnamic acid derivatives in date palm seedlings challenged with Fusarium oxysporum f. sp. albedinis. Proceedings Polyphenols Communications. 22-25 August 2006, Winnipeg, MB, pp. 349-350.

  12. Jaiti, F., Kassami, M., Meddich, A., El-Hadrami, I. (2008). Effect of Arbuscular Mycorrhization on the Accumulation of hydroxycinnamic acid derivatives in date palm seedlings challenged with Fusarium oxysporum f. sp. albedinis. Journal of Phytopathology. 156 (11-12): 641-646.

  13. Kala, C., Gangopadhyay, S., Godara, S.L. (2016). Eco-friendly management of wilt caused by Fusarium oxysporum f. sp. Ciceri in chickpea. Legume Research. 39 (1): 129-134. Doi 10.18805/lr.v0iOF.6789

  14. Mejdoub-Trabelsi, B., Aydi, R.B.A., Nawaim, A., Daami-Remadi, M. (2017). Antifungal Potential of Extracellular Metabolites from Penicillium spp. and Aspergillus spp. Naturally Associated to Potato against Fusarium species. Causing Tuber Dry Rot. Journal of Microbial and Biochemical Technology. 09: 181-190. doi: 10.4172/1948-5948. 1000364.

  15. Munier, P. (1974). Sur l’origine de la datte Deglet-Nour. Fruits. 29 (12) : 823-824.

  16. Ou-Zine, M., Bouhlal, Y., El-Hilali, R., Achbani, E., Haggoud, A., Bouamri, R. (2020). Evaluation of compost quality and bioprotection potential against Fusarium wilt of date palm. Waste Management. 113: 12-19. doi 10.1016/j.wasman. 2020.05.035

  17. Sahampoor, L., Zaker-Travallaie, F., Fani, S.R., Shahbazi, S. (2020). In vitro efficiency of Trichoderma harzianum mutants in biocontrol of Fusarium oxysporum f. sp. radicis-cucumerinum. Journal of Crop Protection. 09(2): 285-300. 

  18. Sajeena, A., Nair, D.S., Sreepavan, K. (2020). Non-pathogenic Fusarium oxysporum as a biocontrol agent. Indian Phytopathology. 73: 177-183. doi https://doi.org/10.1007/s42360-020-00226-x.

  19. Sedra, My H. (1985). Potentiel infectieux et réceptivité de quelques sols de palmeraies ‘à la fusariose vasculaire du palmier dattier (bayoud) causée par Fusarium oxysporum f. sp. albedinis (Kill. et Maire) Malençon. Ph.D. Thesis 3rd. Cycle. Agricultural Sciences. I.A.V. Hassan II, Rabat, Morocco. 88p. 

  20. Sedra, M.H. (2005a). La maladie du Bayoud du palmier dattier en Afrique du Nord : Diagnostic et caractérisation. Proceeding Actes du Symposium International sur le Développement Durable des Systèmes Oasiens, 08-10 March 2005, Arfoud, Morocco. pp. 26-34.

  21. Sedra, M.H. (2005b). Caractérisation des clones sélectionnés du palmier dattier et prometteurs pour combattre la maladie du Bayoud. Proceeding Actes du Symposium International sur le Développement Durable des Systèmes Oasiens, 08-10 March 2005, Arfoud, Morocco. pp 72-79.

  22. Statistica. (2020). Statistiques de production de Dattes en Algérie. https://fr.statista.com/statistiques/991013/production-totale-de-dattes-par-type-algerie/

  23. Touzi, A., (2007). Algerian experience in preserving fragile ecosystems from desertification, a paper presented at “Fifteenth OSCE Economic and Environmental Forum - Part 2: “Key challenges to ensure environmental security and sustainable development in the OSCE area: Land degradation, soil contamination and water management” Prague, 21 - 23 May 2007. Session IV: Challenges to the management of water resources and to countering desertification in the Mediterranean region, Organization for Security and Cooperation in Europe Secretariat.

  24. Vibha, M. (2010). Effect of fungal metabolites and amendments on mycelium growth of Rhizoctonia solani. Journal of Plant Protection Research. 50: 93-97.

  25. Vinita, D., Punia, D. (2016). Nutritional composition of fruit of four date palm (Phoenix dactylifera L.) cultivars grown in Haryana, India. Asian Journal of Dairy and Food Research. 35(4): 331-334. doi 10.18805/ajdfr.v35i4.6635

  26. Zohary, D., Hopf, M. (2000). Domestication of Plants in the Old World (3rd Ed.), Oxford University Press, Oxford, UK.
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    Research Article

    volume 41 issue 3 (september 2021) : 450-454,   Doi: 10.18805/ag.D-304

    Biocontrol of Bayoud Disease (Fusarium oxysporum F. Sp. Albedinis) on Deglet-nour Variety of Date Palm (Phoenix dactylifera L.) in South Western Oases of Algeria

    H
    Hakima Belaidi1,*
    F
    Fawzia Toumi-Benali1
    I
    Ibrahim Elkhalil Benzohra2
    1Laboratoire Ecodeveloppement des Espaces, Department of Environmental Sciences, Djillali Liabes University of Sidi Bel Abbes, Algeria.
    2Department of Phoeniciculture, Experimental Station for Biophysical Environment Bechar (Taghit)-Centre for Scientific and Technical Research on Arid Regions (CRSTRA), Campus Universitaire B.P. 1682 RP, Biskra, Algeria.
    Cite article:- Belaidi Hakima, Toumi-Benali Fawzia, Benzohra Elkhalil Ibrahim (2021). Biocontrol of Bayoud Disease (Fusarium oxysporum F. Sp. Albedinis) on Deglet-nour Variety of Date Palm (Phoenix dactylifera L.) in South Western Oases of Algeria . Agricultural Science Digest. 41(3): 450-454. doi: 10.18805/ag.D-304.

    ABSTRACT

    Background: This work has the objective to biocontrol the bayoud disease of date palm caused by the fungus Fusarium oxysporum f. sp. albedinis (Foa), is the major disease in Algerian palm groves of south western region. This in vivo biocontrol was done on date palm variety Deglet-Nour, using two antagonistic fungi species, Trichoderma harzianum and Aspergillus flavus.

    Methods: Twenty Foa isolates obtained from isolation of the spines carrying the typical symptoms of Bayoud disease were used. Two strains T. harzianum and A. flavus, were isolated from rhizosphere soils of the date palm trees. Using the greenhouse screening test, which was carried out on 3-4 month-old date palm seedlings at the rate of 5 ml of inoculum suspension.

    Result: A statistical analysis showed a significant (P<0.05), difference of Deglet-Nour seedlings reactions against Foa isolates and antagonistic fungi were observed. All date palm trees present susceptibility against Foa until the total mortality of seedlings. The biocontrol test showed that two antagonists showed different reaction, with the complete resistance for seedlings treated by T. harzianum with the mortality rate (rm%) reduced at 100%, while, the second antagonist A. flavus reduced the rm% at 25%. We can apply of these antagonistic fungi to protect our groves contaminated by Bayoud disease and also contain this susceptible commercial variety.

    INTRODUCTION

    The date palm (Phoenix dactylifera L.) is considered a symbol of life in the desert, while it tolerates high temperatures, drought and salinity more than other crop plant species. It is one of the oldest trees from which man has derived benefit and it has been cultivated since ancient times (Zohary and Hopf, 2000). It the only indigenous wild desert plant definitely domesticated in its native harsh environments appears to be the date palm (Zohary and Hopf, 2000 ; Vinita and Punia, 2016). Among the most cultivated and important varieties in Algeria, Deglet Nour, considered as the golden fruit of the oases, means: Deglet: light; Nour: light, is the best variety in the world (Munier, 1974). The Algerian palm groves represent 45% of Deglet Nour variety cultivation (Bouguedoura et al., 2008). The annual production of Deglet-Nour variety of dates is 5670000 quintals (Statistica, 2020), which represents 51,8% of total Algerian date palm production (FAO, 2020). The date palm sector has many problems like sand dunes, salinity, secheresse and seriously damaged by Bayoud disease (Touzi, 2007; Benzohra et al., 2015), which destroyed a half date palm trees of Algerian palm groves in the south-west region (Bouguedoura et al., 2008).


    Bayoud disease caused by the soil fungus Fusarium oxysporum f. sp. albedinis (Foa), is considered as the most destructive fungal disease in south west of Algerian palm groves (Djerbi, 2003). It’s originated from Morocco and its progression continues to advance from the west and central of Algerian palm groves (Benzohra et al., 2015; 2017). The proposal used of prophylactic measures against this disease doesn’t seem able to stop progressive spread of this disease (Benzohra et al., 2017). Other method was used, a genetic control by resistant varieties is therefore another promising way to reduce this constraint, but unfortunately all resistant cultivars haven’t high food quality in date fruits (Sedra, 2005 a,b). It is necessary to protect this variety (Deglet-Nour), against this constraint with an effective method that does not pollute the environment. The biological control remains the best way to combat this constraint to protect this variety (El-Hassni et al., 2007; Jaiti et al., 2006, Jaiti et al., 2008; Kala et al., 2016; Fitrianingsih et al., 2019).         


    In this case, this study has the objective to evaluate the biocontrol capacity of two antagonistic fungi species, Trichoderma harzianum and Aspergills flavus on the incidence of bayoud disease by testing of the important commercial date palm variety, Deglet Nour, but susceptible to Bayoud diseases.

    MATERIALS AND METHODS

    Date palm variety’s seedlings
     
    The date palm variety used in this test named Deglet-Nour, this Algerian variety, has the high quality of date fruit in the word (Munier, 1974).
     
    Fungal material
     
    Twenty isolates of Foa were used in the inoculation test of date palm seedlings, was sampled from contaminated palm groves in Saoura, Gourrara, Touat and M’zab provinces in 2016 (Table 1). The antagonistic microorganisms were obtained from soils of date palm rhizosphere of Bechar region in 2019 (Table 2). These antagonistic fungi used in this study provided by CRSTRA institute (Center for Scientific and Technical Research on Arid Regions, Biskra, Algeria), from Laboratory of Plant Pathology and Protection, of Experimental Station of Biophysical Environment, Naama, Algeria. The isolates were conserved in PDA cultures (For each 1000 ml of distillated water: 200 g potato filtrate; 15 g glucose; 20 g agar-agar; pH=5,5), with temperature of conservation at 04°C and incubation at 23°C (El-Hassni et al., 2007).
     

    Table 1: Foa isolates with its origin.


     

    Table 2: Informations of antgonistic fungi strains’ species used in ‘in vivo’ biocontrol test against bayoud disease.


     
    Preparation of inoculum
     
    Date palm seedlings of Deglet-Nour variety of 2-3 months-old, were obtained by pre-germination of date palm seeds (Fig 1), after disinfection with Hypochlorite of Sodium at 2%, then rinsed with sterile distilled water three successive times (Sedra, 2005b). The date palm seeds were placed in Petri dishes containing the cotton soaked with sterile distilled water and then stored in the oven at a temperature of 23±2°C (El-Hassni et al., 2007). After 20 days of pre-germination (Fig 1), the seeds were transplanted into pots containing sterile peat for two months (Sedra, 1985).
     

    Fig 1: Pre-germinatings seeds of Deglet-Nour variety used for test of biocontrol.


     
    The in vivo biocontrol test was performed using the mixture ‘Foa inoculum + antagonistic fungus’ and two controls, one for seedlings treated with Foa inoculum alone and other without Foa inoculum. Four seedlings were used per treatment and repetition. After inoculation tests, the seedlings were conserved in small greenhouse at 25±2°C with 16 h of photoperiod.
     
    Rating scale
     
    The mortality rate of date palm seedlings was based on rate of mortality and calculations of dead plants as compared to all the plants inoculated (Sedra, 2005b). The mortality rate (mr) formula was as follows:

    Statistical analysis
     
    The experiment design was randomized complete blocks. Four replicates (number of seedlings tested), per treatment were taken and each whole experiment was performed independently. All collected data were submitted to ANOVA analysis using XLStat software 9 (AddinSoft, USA Ltd.) and the significance of differences was recorded at P<0.05, by Newman-Keuls test.

    RESULTS AND DISCUSSION

    The results of statistical analysis showed significant differences (P0.05), between the date palm varieties and also between two antagonistic microorganisms (Table 3). The results showed a high level of reduced mortality rate (mr) due to two antagonists of incidence of Bayoud disease on deglet-Nour variety seedlings compared to the control.   The control (direct inoculation with Foa alone), presented a very high mortality rate up to 100% (Table 3), while the control without Foa isolates inoculums had no typical symptoms of bayoud (Fig 2). Absences of symptoms were observed on seedlings inoculated by the antagonistic fungus T. harzianum with no mr (%). The second antagonistic fungus A. flavus could reduced the mr at 25% of seedlings tested (Table 3). In comparison between these two antagonistic species, we confirm that T. harzianum was more efficient than A. flavus.
     

    Fig 2: Reactions of Deglet-Nour variety’s seedling against Bayoud disease in presence of two antagonistic fungi species. Foa: Fusarium oxysporum f. sp. albedinis; antagonistic fungi species: T. harzianum: Trichoderma harzianum, A.flavus.: Aspergillus flavus; *: date palm varieties names.


     

    Table 3: Mortality rates and reaction of Deglet-Nour variety seedlings of date palm in the biocontrol test against bayoud disease.


     
    Our study showed that the antagonistic fungal species tested have significant biocontrol activity against Bayoud disease when tested in vivo. The mortality rate (mr) varied significantly ascompared tocontrol, when compared with two antagonistic fungi, T. harzianum was more efficient with mr=0%, than A. flavus with mr =25%.
           
    Many reports have been published about the importance of biocontrol as the best way to combat Fusarium wilt (Sajeena et al., 2020; Cheng et al., 2019; Ou-Zine et al., 2020). The importance of antagonistic fungi like T. harzianum as biofungicide against Fusarium wilt of many crops has been reported by Kala et al., (2016), including Fusarium wilt of chickpea (Cicer arietinum L.) caused by Fusarium oxysporum f. sp. ciceris, by Sahampoor et al., (2020), on Fusarium wilt of cucurbits caused by Fusarium oxysporum f. sp. radicis-cucumerinum and also by Fitrianingsih et al., (2019), on Fusarium wilt of tomato (Lycopersicum esculentum L.) caused by the soil fungus Fusarium oxysporum f. sp. radicis-lycopersici.
     
    The antagonistic fungus Aspergillus flavus was also reported by many authors effective against Fusarium wilt diseases. Among these, Fusarium oxysporum f. sp. melonis of melon (Cucumis melo L.), Fusarium oxysporum f. sp. vasinfectum of Cotton (Gossypium herbaceum L.), (Dong and Cohen, 2002), Fusarium graminearum of cereals (Mejdoub-Trabelsi et al., 2017) and Rhizoctonia solani (Vibha, 2010) and Fusarium sambucinum of potato (Solanum tuberosum L.), (Aydi et al., 2015).
     
    It can therefore be concluded that two antagonistic fungi species (T. harzianum and A. flavus), can enhance the ability to of host to resist bayoud disease in Deglet-Nour variety under in vivo biocontrol tests in palm groves in Algeria.

    CONCLUSION

    This study had the objective to evaluate the capacity of in vivo biocontrol on incidence and aggressiveness of Bayoud disease caused by Fusarium oxysporum f. sp. albedinis (Foa), which is  the major constraint of date palm cultivation in south-western palm groves of Algeria. This biocontrol was realized by use of two antagonistic fungal species Trichoderma harzianum and Aspergillus flavus, against the important variety of date palm in Algeria named Deglet-Nour. The results showed significant differences between two antagonistic fungi as well as the control (inoculation with and without Foa strains mixture inoculum). In comparison between these two antagonistic species, we can say that T. harzianum is more efficient than that species A. flavus, with mr % values  of 0%, 25%, respectively.
     
    The use of biological agents manage disease in the susceptible varieties like Dglet-Nour against Bayound provide an ecologically friendly alternative to pesticides to control the disease. This strategy has more significance against soil-borne pathogens such as Fusarium wilt in palm groves of south-west region of Algeria.

    ACKNOWLEDGEMENT

    We would like to thanks the DGRSDT (Direction Générale de la Recherche Scientifique et du Développement Technologique, Algiers, Algeria), affiliated with the Ministry of High Education and Scientific Research of Algeria, for its funding support which facilitated us to realize this work in  the  Social – Economical project. 

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