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عزل وتشخيص البكتيريا Bacillus thuringiensis ودراسة تاثيرها في بعض المقاييس الحياتية لذبابة ثمار القرعيات (Diptera : Tephritidae) Dacus ciliatus == Isolation And Identification of Bacillus Thuringiensis And Studying Its Effects On Some Biological Parameters For Dacus Ciliatus (Diptera : Tephritidae)
Author name:
ادريس صالح جليل
Supervisor name:
عدنان نعمة عبد الرضا العزاوي
General topic:
Biology
Specific topic:
Microbiology
Degree:
Master
University:
University of Diyala - College Of Education For Pure Sciences - Department Of Biology
Language:
Arabic
University location:
Diyala
First pages:
24T2866 - p.pdf
Abstract:
تضمنت هذه الدراسة عزل البكتيريا Bacillus thuringiensis من مصادر بيئية مختلفة في محافظة ديالى وتشخيصها بالطرق الكيموحيويه والوراثية والجزئية، اذ جمعت 51 عينة من ترب مختلفة وهي(زراعية وحديقة وغير زراعية) وكذلك من الماء، من خمسة اقضية في محافظة ديالى (بعقوبة | This study included the isolation of pathogenic bacteria Bacillus thuringiensis from different environmental sources in Diyala province, diagnosis biochemical and genetic partial ways, Mar 50 samples collected from different soils, namely, (agricultural soils and garden soils and non - agricultural soils) as well as from the water, from five districts in the province of Diyala (Baquba, Khalis, Muqdadiyah, Khanaqin, Balad Ruz) for the period from 1 \ 9 \ 2014 to 20 \ 11 \ 2014. Showed the results of biochemical diagnosis and the presence of four samples and by percent 8% It was of a negative for transplantation bacterial and 46 sample growth by percent 92% It was of a growth positive for transplantation bacterial and found that 41 samples of which by percent 82% belong to the genus Bacillus and the species of Bacillus thuringiensis and the percentage of bacteria isolated of agricultural soils 94% and soils garden 93% and water 75%, and soils non - agricultural 44%. The pathogenic bacteria Bacillus thuringiensis of positive bacteria a gram dye and spore forming and The most of the natural habitat this bacterium is soil and it is capable of producing the diversified varieties of crystalline proteins with insecticide property. All isolates were tested for this bacteria that which were able growth on the media selective Acetate Sodium Selective and the production of insecticidal crystalline proteins (ICP) When staining by dye Coomassie brilliant blue (CBB) to conduct genetic partial diagnosis using several extraction (Mini gDNA Bacteria kit) processed by Bioneer company was the purity of DNA extracted ranging between (20.1 - 1.22) for all selected isolates, have been diagnosed with the bacteria at the level of genus and species Using the gene 16S rDNA with a molecular weight of 380 base pairs where the results showed polymerase chain reaction (PCR) that all isolates contain this gene and the same molecular weight which is 380 base pairs, also was diagnosed isolates containing the gene component crystalline proteins which cry a e molecular weight 200 base pairs using primer a specialist for this gene in polymerase chain reaction showed the results of gene doubled that all isolates contained the gene and the same molecular weight of 200 base pairs. After making sure of pathogenic bacteria as belonging to the genus Bacillus and species Bacillus thuringiensis according to their chemical diagnostics biochemical and diagnostic genetic partial, attended different concentrations They (1, 2, 3, 4, 5) g/L of thebio preparation for these bacteria to study their impact on the different roles of Cucurbits fruit fly Daus ciliatus. Has been tested the effect of different concentrations of bio - preparation of these bacteria isolated from soil and water on the roles of this insect which eggs and larvae and pupa and AdultsThe results of test the effect of bio - preparation of these bacteria on the eggs of age (1 - 2) Since the highest rate of hatchling (3.2) and by (89%) at a concentration of 1 g / l of bio - preparation and lowest hatchling (0.6) and by (19%) at a concentration of 5 g / L of bio - preparation comparison with control (3.2) and by (96%) for the isolated bacteria from soil Since the highest rate of hatchling (3.1) and by (92%) at a concentration 1 g / L of bio - preparation and lowest hatchling (2.1) and by (62%) at a concentration of 5, 4 g / L of bio - preparation comparison with control (3.2) and by (96%) for isolated bacteria from water, and on The last larval phase, The highest the value of the rate of highest (2.0) and by (99%) at a concentration of 5 g / L of bio - preparation and The less the value of the rate of loss (0.5) and by (26%) at 1g / L of bio - preparation concentration compared with the control treatment (0.07) and by (%3) for isolated bacteria from soil.reached its highest value the loss rate (1.2) and by (57%) at a concentration of 5 g / L of bio - preparation and less the value of the loss rate (1.0) and by (6%) at a concentration 1 g / L of bio - preparation comparison with control (0.07) and by (3%) for the bacteria isolated from water and on pupa age(6 - 7) day, reaching the highest rate of emergence of pupa (3.2) and by (92%) at a concentration of 1 g / L of bio - preparation and less the emergence of pupa rate (0.4) and (13%) at a concentration of 5 g /L of bio - preparation comparison with control (3.2) and by 96% for bacteria isolated from soil reaching the top of the emergence of pupa rate (3.1) and by (92%) at a concentration of 1 g / L of bio - preparation and less the emergence of pupa rate (2.1) and by (62%) at a concentration of 5 g / L of bio - preparation comparison with control (3.2) and by (96%) for bacteria isolated from water. and on kill adult, The highest killing rate for males (1.6) and by (32%) and females (1.5) and (30%) at a concentration of 5 g / l of bio - preparation and less killing of the male rate (0.5) and (10%) and females (0.4) and by (13%), comparison with control males (0.1) and by (2%) and females (0.1) and by (2%) for the isolated bacteria from soil. The highest kill the male rate (0.8) and by (32%) and females (0.7) and (30%) at a concentration of 5 g / L of bio - preparation, and less killing of the male rate (0.2) and (4%) and females (0.4) and by (13%) at a concentration of 1 g / L of bio - preparation comparison with control males (0.1) and by (2%) and females (0.1) and by (2%)for the isolated bacteria from water and one female productivity of egg reached its highest production rate eggs 84.5 eggs per female at a concentration of 1 g / L of bio - preparation, and less egg production rate of 54.5 eggs per female at a concentration of 5 g / L of bio - preparation comparison with control eggs productivity rate of 87.5 egg female for isolated bacteria from soil reaching its highest egg production rate (85%) eggs per female at a concentration of 1 g / L of bio - preparation, and less egg production rate (79.5%) eggs per female at a concentration of 5 g / L of bio - preparation, compared with treatment control productivity rate of whites (85.5%) eggs per female for isolated bacteria from the water
