الفهرس 
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Abstract
The present study was conducted during two successive seasons 2017 and 2018, on Flame seedless grapevines cultivar (Vitis vinifers L.). The experimental work of this study included two main separated parts, as follows:
1- First part, Field trails:
This part was conducted under field conditions. This part includes two separated experiments, each one were carried out on thirty two uniforms in vigour Flame Seedless grapevines. The selected vines load was 72 eyes, for all the selected vines, on the basis of six fruiting canes X ten eyes plus six renewal spurs X two eyes. The vines grown under two different types of soil as follows:
The first experiment: carried out under will drained clay soil. However, the water table depth is not less than two meters. The select orchard was located at Com El-Arab village Matay district, Minia Governorate – Egypt. The vines were 20 years old and irrigate by surface system using Nil river water. The vines are planted at 2.5 x 3.0 m apart, pruned during the first week of January in the two seasons using cane pruning method with the assistance of Gable supporting system were used.
The second experiment: carried out on 32 uniforms in vigour Flame Seedless grapevines grown under new reclamation desert land conditions, located at the border of Eastern desert rood, facing of Matay district, El-Minia Governorate - Egypt. Whereas, the
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selected vines were 11 years old and irrigate by drip irrigation system, using water supply from underground well with pressure and volume controllers. The selected vines are planted at 2.0 x 3.0 m apart, pruned during the last week of December in the two seasons using cane pruning method with the assistance of Gable supporting system.
For ache type of soils (sandy or clay), seven treatments from soil single and combined inoculation with the three microorganisms namely: Arbuscular Mmycorrhizal fungi (AMF), Azospirillum brasilense bacteria (AZSB), and Azotobacter chrococcum bacteria (AZBB), in addition to the control treatment:
o Control (un-inoculated vines).
o Inoculation with Arbuscular Mmycorrhizal fungi (AMF).
o Inoculation with Azospirillum brasilense bacteria (AZSB).
o Inoculation with Azotobacter chrococcum bacteria (AZBB).
o Inoculation with Arbuscular Mmycorrhizal fungi combined with Azospirillum brasilense bacteria (AMF + AZSB).
o Inoculation with Arbuscular Mmycorrhizal fungi combined with Azotobacter chrococcum bacteria (AMF + AZBB).
o Inoculation with Azospirillum brasilense bacteria combined with Azotobacter chrococcum bacteria (AZSB + AZBB).
o Inoculation with Arbuscular Mmycorrhizal fungi combined with Azospirillum brasilense bacteria and Azotobacter chrococcum bacteria (AMF + AZSB + AZBB).
Each treatment was replicated four times, one vine per each.
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Microorganism strains: The Fungi and bacterial strains used in these experiments (AMF, AZSB, and AZBB, as well as their mixtures) were kindly isolated and propagated in Laboratory of Microbiology, Minia University, Egypt. Strains of AZSB or AZBB were grown on Doberiner medium. Strains were grown in liquid medium on a rotary shaker at 30 °C and 120 rpm, then the culture were added to the vines, three times/year, at a rate of 200 ml per vine, however each ml contain 108 cell of Azospirillum or Azotobacter bacteria. AMF was developed on onion plants roots, and so the onion soil added to the vines in order to 200g/vines. However, each 1 gram contained 108 spores.
Randomized complete block design (RCBD) was followed, where the experiment consisted of eight treatments, each treatment was replicated four times, one vine per each. The obtained data were tabulated and significantly analyzed. The differences between the means were compared using new L.S.D. test at 5% level of probability.
2- Second part, soilless culture:
The experimental work of this part was conducted during two successive seasons (2017 and 2018) on Flame seedless (Vitis vinifera L.) seedlings, under controlled conditions in a Laboratory. However, the temperature adjusted to 25 ± 3C°, relative humidity ranged between 75 – 85 % and 14 hours exposure to light.
Plant material: The used seedlings of the Flame seedless cultivar were one year old and own rooted. In January, the seedlings were
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pruned leaving two spurs/plant, each spur containing four eyes, then they cultivated on sandy soil and irrigated with the referenced nutrient solution until the half of February, then the plants were transported to the hydroponic solution culture contained the different microorganisms used.
Nutrient solution: Standard nutrient solution supplemented with the nutrients requirements of grapevines was prepared. The nutrient solution contained Macro-nutrients (meq/L): 8.5 NO3; 1.0 H2PO5; 1.3 SO4; 1.0 NH4; 2.1 K; 6.7 Ca; 2.0 Mg and Micro-nutrients (meq/L): 5.9 Fe; 2.0 Mn; 0.05 Mo; 1.50 B; 0.5 Zn; 0.25 Cu. The solution was reached with the microorganisms (AMF, AZSB and AZBB single or combined inoculations).
Microorganisms strains: The AMF, AZSB, and AZBB Strains, as well as their all possible combination were kindly isolated and propagated at Laboratory of Microbiology, Minia University, Egypt. Strains of AZSB were grown on Doberiner medium but AZBB was grown on nutrient broth medium. Then, the culture were added to the nutrient solutions, one time weekly, at a rate of 20 ml per pot, each ml contain 108 cell of AZSB or AZBB. However, AMF was developed on onion plants roots, then the onion soil were extracted in water solutions, the solution was added weekly to pots in order of
20 ml/pot. However, each 1 mal contained 108 germs.
Complete randomized design (CRD) was followed, where the experiment consisted of eight treatments, each treatment was
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replicated five times, one pot per each and each pot occupied by two seedlings.
The overall aims of these experiments were to improve understanding of the influence of Flame Seedless grapevines seedlings inoculation with AMF, AZSB, and AZBB in single or combined application on some vegetative growth characteristics, leaf chemical composition and yield as well as fruit physical and chemical characteristics of Flame Seedless grapevines grown in clay, sandy and soilless conditions.
The most important results obtained during two experimental seasons could be summarized as follows:
1- Vegetative growth characteristics: All studied vegetative growth characters (i.e. main shoot length (cm), number of leaves/shoot and leaf area (cm2)) were significantly stimulated in response to single or combined inoculation with AMF, AZSB, and AZBB under field or soilless culture conditions, in both experimental seasons. However, under field conditions, in clay soil AZSB+AZBB co-inoculation remain more effectiveness on all vegetative growth parameters than AMF+AZSB or AMF+AZBB co-inoculation. While, AMF + AZBB co-inoculation present superior values of shoot lengths (cm) and leaf area (cm2) than AMF+AZSB or AZSB+AZBB co-inoculations. Furthermore, under soilless culture condition the co-inoculation of AMF + AZSB shows a superior effect on all vegetative growth parameters compared to AMF+AZBB or
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AZSB+AZBB co-inoculations during the two experimental seasons. The highest vegetative growth parameters of Flame seedless vines or seedlings were obtained from the vines inoculated the mixture of the three microorganisms in combination (AMF+AZSB+AZBB), under clay and sandy soils conditions as well as in soilless culture.
2- Leaf pigments contents: Inoculated Flame Seedless grapevines with single or combined AMF, AZSB, and AZBB strains was associated with remarkable promotion on chlorophylls a & b and total chlorophylls (mg/100g F.W.) as well as total carotenoids pigments, rather than un-inoculated vines.
Under field or soilless culture conditions, the obtained data showed also that all combined inoculations present higher leaves pigments than the single or un-inoculated ones. Furthermore, the vines or the seedlings received the mixture of the three microorganisms present higher and significant chlorophylls (a, b, and total chlorophyll) and total carotenoids in their leaves than the other inoculations or un-inoculated ones, during the two experimental season. On the opposite side, untreated vines or seedlings present the lowest pigments content in their leaves during the two experimental seasons, under field and soilless culture conditions.
3- Leaf mineral contents: A promotion effect was observed on Flame Seedless leaves contents of macro nutrients (N, p, K and Mg) as well as micro nutrients (Fe, Zn and Mn) when the vines inoculated with AMF, AZSB, and AZBB under clay and sandy soils
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as well as soilless culture over the check treatments. Concerning the effect of individual inoculations with the three examined microorganisms, the vines or the seedlings inoculated with AMF preset higher P, K, Mg, Fe, Zn and Mn (contents) in their leaves than those inoculated with AZSB or AZBB individually or un-inoculated ones. While, the vines inoculated with AZSB alone present higher N% than those inoculated with AMF or AZBB under field condition or soilless culture. Co-inoculation with AMF+AZSB was more effectiveness than AMF+AZBB or AZSB+AZBB on leaves P, K, Mg, Fe, Zn and Mn (contents) in both field and soilless culture. While, co-inoculation with AZSB+AZBB gave higher and significant N% rather than the two other co-inoculations. Furthermore, the vines or the seedlings inoculated with the mixture of the three microorganisms (AMF+ AZSB + AZBB) in combinations present the highest values of mineral nutrients in their leaves during the two experimental seasons, in both field and soilless cultures.
4- Yield and its components: Inoculated Flame Seedless grapevines with AMF, AZSB, and AZBB under both clay and sandy conditions had no considerable promotion on clusters number per vine in the first season of this study. However, the cluster weight was remarkably improved. In the second experimental season, a remarkable promotion in both of the clusters number per vine, cluster weight as well as yield (kg) per vine were observed. However, he highest clusters number/vine, cluster weight, and yield/vines of Flame Seedless grapevines grown under clay or
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sandy soils conditions were obtained from the vines inoculated with AMF+AZSB+AZBB in combinations. Then, for improving yield (kg)/vine as well as cluster weight (g) and clusters number/vine in clay or sandy soils it was preferable to inoculate the vines with a mixture of AMF+AZSB+AZBB in combinations.
5- Berries Physical properties: Under clay and sandy soils, there was an obvious effect on physical characteristics of Flame Seedless grapevines berries due to inoculate the vines with AMF, AZSB, and AZBB, as a single or in combinations, over un-inoculated vines, during the two experimental seasons. This was appeared in terms of increasing berry weight, Berry volume, and beery dimensions. However, the co-inoculation AMF+AZBB under clay soil conditions and AMF+AZSB under sandy soil condition remain more effectiveness than the other co-inoculations, during the two experimental seasons. Furthermore, the vines inoculated the mixture of the three microorganisms (AMF+AZSB+AZBB) in combination present the highest berry weight, berry volume, and berry dimensions, under clay and sandy soil conditions in both experimental seasons.
6- Berries Chemical properties: Inoculated Flame Seedless grapevines grown under clay and sandy soils conditions with AMF, AZSB, and AZBB had considerable promotion on the fruit chemical properties in terms of increasing TSS%, Reducing sugars%, TSS/acid ratio, total anthocyanins, and decreasing total acidity%, rather than un-inoculated vines. Also, it is clear from the obtained
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that the co-inoculation AMF+AZSB remain more effectiveness than AMF+AZBB or AZSB+AZBB co-inoculations in both clay and sandy soil. The best results concerning the chemical properties of berries were obtained from the vines inoculated with the mixture of AMF+AZSB+AZBB in combination under clay and sandy soil conditions, during the two experimental seasons.
Conclusion: from the results of the present study it is clear that inoculated Flame Seedless grapevines grown in clay and sandy soils as well as under soilless conditions with AMF, AZSB, and AZBB as a single or combined inoculations may play a beneficial role in enhancing vegetative growth aspects, leaves pigments content, leaves mineral contents and yield as well as berries physical and chemical properties. However, the inoculations with the mixture of AMF+AZSB+AZBB present the highest vegetative growth parameters, leaves main pigments content and nutritional status, yield and fruit quality.
It is strongly recommended to inoculate Flame Seedless grape vines grown under the experiment conditions (Minia Governorate) in clay and sandy soils and resembling conditions, also under soilless conditions with the mixture of AFM+AZSB+AZBB three times during the growth season, in order to improve the different vegetative growth, leaves chemical component and improving the yield and berries quality.