الفهرس 
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Abstract
Grapevine is one of the most important fruit crops grown in tropical, subtropical and cold areas all over the world. It is used in fresh food for industrial and medical purposes. Grapevine is the second fruit crop after citrus in Egypt. The total cultivated area (198934) feddan produces 1686169 tons of grapes while the area planted with grapevine in El-Nobaria Province accounts for around 47% of all the land planted with grapes in Egypt. In Egypt and all over the world grapevine cultivation is subject to attack by root rot disease caused by soil borne fungi i.e. Fusarium spp., Macrophomina phaseolina, Rhizoctonia solani, Pythium ultimum and Lasiodiplodia theobromae causing decline in vegetative growth and fruit quality and quantity.
The present work aimed to find out a control program of root rot disease of grapevine using preparation of metallic nanoparticles by biological, physical and chemical methods, testing against fungal pathogens and their effect on root rot disease incidence of grapevine and plant growth.
The results obtained can be summarized as follows:
1. Survey of root rot disease on grapevine trees in El-Nobaria Province, El-Beheira Governorate in Egypt. The results indicated that high percentage of diseased grapevine tree and disease severity was increased from 2019 to 2022 on flame seedless followed by crimson then superior during summer seasons.
2. Isolated (117 fungal isolate) from the root rot tissue of trees grapevine showed symptoms of infection by diseases of the roots of grapevine from some provinces and found that Fusarium spp. was the most frequent fungi followed by Lasiodiplodia theobromae and then the fungus Macrophomina phaseolina and the following percentages (37.6%, 31.6% and 22.2%) respectively, while R. solani was the least frequent (8.6%). On the other hand highly frequency of fungi was recorded in flame seedless followed by crimson and then superior cultivars respectively.
3. Four types of fungal infection types were recorded in El-Nobaria Province, El-Beheira Governorate due to Fusarium spp., Lasiodiplodia spp., Macrophomina phaseolina and Rhazoctoina solani. First infection type caused by each individual fungus with highly frequency by Fusarium spp. followed by Lasiodiplodia spp. Second infection type caused by Fusarium spp. + Lasiodiplodia spp. followed by Fusarium spp. + M. phaseolina and then Lasiodiplodia spp.+ M. phaseolina. Third infection type was caused by Fusarium spp. + Lasiodiplodia spp.+ M. phaseolina. Fourth infection type were recorded between four fungi Fusarium spp.+ Lasiodiplodia spp.+ M. phaseolina + R. solani which only recorded on flame seedless cultivar in this study. Fusarium spp. was the most fungi associated with all infection types at all grapevine cultivars in this study followed by Lasiodiplodia spp.
4. Isolates of fungi were identified morphologically according on the basis of cultural traits of mycelial growth, pigmentation and microscopic observations of macroconidia, microconidia, sclerotia and their morphological characteristics. Isolates of fungi were identified according to characterizations of molecular biology based on the internal transcribed spacer 1(ITS1) sequencing and conserved in GenBank with accession numbers from (ON037457.1 to ON037474.1).
5. The pathogenicity test was tested on eighteen isolates of fungi were isolated from different grapevine cultivars from El Nobaria province. The test showed that fungal isolate of Fusarium solani (kh6) was the most fungal isolate recorded high and significant root rot percentage and disease severity on grapevine plants followed by Lasiodiplodia theobromae (kh18), Fusarium oxysporum (kh17) and Macrophomina phasiolina (kh13). Meanwhile, the fungus Fusarium solani (kh5) moderate in the ability to causing root-rot disease of grapevine then Lasiodiplodia crassispora (kh7) was the least capable of causing the disease.
6. This is first record that Fusarium chlamydosporum, F. brachygibbosum, F. ipomoeae, Lasiodiplodia crassispora and L. exigua as a causal organisms causing root rot disease of grapevine in Egypt.
7. The effect of different fungal isolates on the morphological characteristics i.e. plant height of shoot, root length, fresh and dry weight of shoot and root and root size of grapevine plants flame seedless cultivar was studied. It was found that all tested isolates resulted in reduction in the morphological characteristics of grapevine seedlings compared to control. Fusarium solani (kh6) the highly pathogenic isolate was reduced the morphological characteristics of grapevine seedlings compared to other isolates.
8. Silver (silver nitrate), silicon (sodium silicate), copper (copper sulfate pentahydrate) and selenium (sodium selenite) were used for preparation nanoparticles by various methods such as biological, chemical and physical. Transmission electron microscopy (TEM) used to visualize shape and sizes of nanoparticles have predominantly shown spherical shape structures and average diameter ranged of biological, chemical and physical synthesized at 7-38, 5-46, 7-62 nm respectively.
9. The average diameter, the size distribution and zeta potential of nanoparticles were measured by using a particle size analyzer, biological nanoparticles, silver 12.69 nm, silicon 28.83 nm, copper 37.95 nm and selenium 37.45 nm, chemical nanoparticles, silver 34.15 nm, silicon 27.28 nm, copper 45.48 nm and selenium 54.19 nm, physical nanoparticles, silver 50.51 nm, silicon 72.26 nm, copper 90.40 nm and selenium 67.67 nm.
10. The effect of use some nanoparticles i.e. silver nanoparticles at 0, 5, 10 and 20 ppm and copper, silicon and selenium nanoparticles at 0, 25, 50 and 100 ppm prepared by biological, chemical and physical methods were tested their antifungal properties on mycelial linear growth of highly aggressive fungal isolates causing root-rot of grapevine. The results showed that significantly reduced mycelial linear growth of Fusarium oxysporum, F. solani, Lasiodiplodia theobromae and Macrophomina phaseolina isolates than the control. The antifungal effect was increased by increasing concentrations. The results indicated that 20 ppm of silver nanoparticles and 100 ppm of silicon, copper and selenium nanoparticles were the best concentration tested which significantly and completely inhibited (100%) linear growth of highly aggressive fungal isolates causing root-rot of grapevine.
11. Biological nanoparticles were prepared by isolate of F. solani, Silver nanoparticles were the best nanoparticles reduced linear growth of four highly pathogenic fungal isolates followed by silicon nanoparticles. Meanwhile, selenium nanoparticles were recorded the lowest antifungal effect of linear growth of fungal isolates. Chemical nanoparticles, Silicon nanoparticles were the best nanoparticles reduced linear growth of four highly pathogenic fungal isolates followed by silver nanoparticles. Meanwhile, selenium nanoparticles were recorded the lowest antifungal effect of linear growth of fungal isolates. Physical nanoparticles, Silver nanoparticles were the best nanoparticles reduced linear growth of four highly pathogenic fungal isolates followed by silicon nanoparticles. Meanwhile, selenium nanoparticles were recorded the lowest antifungal effect of linear growth of fungal isolates.
12. The effect of use some chemical components and fungicide i.e. silver nitrate at 0, 50, 100 and 200 ppm and copper sulfate pentahydrate, sodium silicate, sodium selenite and fungicide Topsin (M-70) at 0, 200, 400 and 600 ppm were tested on the average mycelial linear growth of highly pathogenic fungal isolates caused root-rot disease of grapevine i.e. Fusarium solani, F. oxysporum, Lasiodiplodia theobromae and Macrophomina phaseolina. The results showed that significantly reduced mycelial linear growth of four highly pathogenic fungal isolates caused root-rot disease of grapevine than the control. The antifungal effect was increased by increasing concentrations. Fungicide Topsin (M-70) were the best reduced linear growth of four highly pathogenic fungal isolates followed by sodium silicate. Meanwhile, sodium selenite were recorded the lowest antifungal effect of linear growth of fungal isolates.
13. The effect of use some nanoparticles phytotoxicity on grapevine plants, cutting of healthy grapevine were prepared and dipped for 24 hours in each solution of different nanoparticles prepared by biological, chemical and physical methods i.e. silver nanoparticles at (5, 10 and 20 ppm), copper, selenium and silicon nanoparticles at (25, 50 and 100 ppm). The results showed that were significantly increased the growth rate of grapevine cuttings, the length of the vegetative growths and the chlorophyll content compare than the control. Biological nanoparticles were the best nanoparticles increased the growth rate of grapevine cuttings, the length of the vegetative growths and the chlorophyll content followed by chemical nanoparticles. Meanwhile, physical nanoparticles were recorded the lowest. No phytotoxicity effect on grapevine plants of nanoparticles used in this study.
14. In green house experiments, effect of nanoparticles against highly pathogenic fungal isolates causing root rot on grapevine(cv. Flam seedless) were prepared by biological, chemical and physical methods i.e. silver nanoparticles at 40 ppm concentration, Meanwhile, copper, silicon and selenium nanoparticles and fungicide Topsin (M-70) at 200 ppm were used as soil drench at cultivation time. The results showed that soil drench with silicon biological nanoparticles and silver chemical nanoparticles were the best and significantly treatments completely suppress (100%) of root-rot disease incidence and disease severity of grapevine plants. Meanwhile, soil drench with selenium nanoparticles was the least effect in this respect.
15. Effect of nanoparticles on morphological characters of grapevine (cv. Flam seedless) under artificial infestation by causal pathogens. The results indicated that all different treatments were significantly increased including all morphological characters tested i.e. length of grapevine plants shoot and root, fresh and dry weight of shoot and root as well as root size than in the control, the best and significantly treatments were increased most morphological characters of grapevine plants were silicon biological nanoparticles treatment followed by silver biological nanoparticles. Selenium nanoparticles were the least effect in this respect. In general, biological nanoparticles of various materials were significantly effective increased shoot length and fresh weight as well as length and root size of grapevine plants than it’s prepared by chemical and physical methods respectively.
16. Effect of nanoparticles on the amount of chlorophyll was studied in the grapevine leaves (Cv. Flam seedless) under artificial infestation by causal pathogens. The results showed that all the treatments were significant increase in the amount of chlorophyll in the leaves of grapevine plants compared with control. The best and effective treatment increased chlorophyll content of grapevine leaves were silicon biological nanoparticles and the lowest treatment was selenium chemical nanoparticles.
17. Effect of nanoparticles on chitinase and oxidative enzymes activity in roots of grapevine plants. The results showed that all treatments were significantly increased activities of chitinase and oxidative enzymes tested i.e. peroxidase and polyphenol oxidase in roots of grapevine plants at three month after cultivation compare the control. The best and significantly treatment was increased activities of chitinase and oxidative enzymes was silicon biological nanoparticles its increased over production than the control of peroxidase, polyphenol oxidase and chitinase 527%, 380% and 580% respectively. Generally indicated that the highly activity of peroxidase enzyme were recorded than polyphenol oxidase and chitinase.
18. Effect of nanoparticles on total count of pathogenic fungi on grapevine plants rhizosphre under artificial infestation by causal pathogens. All different treatments soil drench with nanoparticles prepared by biological, chemical and physical methods i.e. silver, copper, silicon and selenium nanoparticles and fungicide Topsin (M-70) were significantly reduced total count of causal organisms compare than the control. Silicon biological nanoparticles treatment was the best and significantly reduced total count of Fusarium spp. in the rhizosphre of grapevine plants followed by silver biological nanoparticles.
19. Under field conditions, evaluation different nanoparticles prepared by biological, chemical and physical methods (silver, copper, silicon and selenium nanoparticles) and fungicide Topsin M 70 on root rot incidence of grapevine were tested, in heavy natural field affected with root rot disease on grapevine trees. Two separate field experiments soil drench with two doses at March and May during 2023 by 3 liter around each tree of grapevine plants Flam seedlss and Crimson cultivars were carried out in Um EL- Muminin, EL- Wafa walaml village’s, EL- Nobaria province, EL- Beheira, Governrate, Egypt.
20. Experiment 1, different nanoparticles prepared by biological, chemical and physical methods (silver, copper, silicon and selenium nanoparticles) and fungicide Topsin (M-70) on grapevine plants Flam seedlss cultivar were significantly reduced percentage of root rot disease, disease severity and increased yield of grapevine plants than the control. Soil drench with silicon biological nanoparticles was the best and significantly treatments than all another treatments followed by silicon chemical and physical nanoparticles. Meanwhile, soil drench with selenium nanoparticles was the least effect in this respect.
21. Experiment 2, different nanoparticles prepared by biological, chemical and physical methods (silver, copper, silicon and selenium nanoparticles) and fungicide Topsin (M-70) on grapevine plants Crimson cultivar were significantly reduced percentage of root rot disease, disease severity and increased yield of grapevine plants than the control. Soil drench with silicon biological nanoparticles was the best and significantly treatment than all another treatments followed by silicon chemical nanoparticles and silver biological nanoparticles. Meanwhile, soil drench with selenium nanoparticles was the least effect in this respect.