Defence development of biopesticide for the control of root pathogenic fungi in chickpea using plant growth promoting rhizobacteria by shazia shahzaman
Thesis titled "Development of Biopesticide for the control of Root Pathogenic Fungi in Chickpea using Plant Growth Promoting Rhizobacteria ".
• Supervised by Prof. Dr. M. Inam-ul-Haq.
• Isolation and Characterization of Rhizbacterial isolates from Rawalpindi District
• Utilization of PGPR antagonistic potential in the form of biopesticide formulation against Fungal Root Infecting Pathogens.
• The Developed formulations with best shelf life and Rhizobacterial viability were evaluated for their efficacy under open field conditions for disease control and plant growth enhancement.
Bio efficacy of pseudomonas fluorescens isolated from chickpea fields as plan...Shazia Shahzaman
Chickpea is an economically important food crop, which is subjected to infection by a host of fungal, viral and bacterial pathogens. Thirty isolates of Pseudomonas fluorescens were isolated from the rhizosphere of Chickpea fields. These were tested against F. oxysporum in dual culture method. Among these, four (Pf 1, Pf 3, Pf 5 and Pf
8) isolates were showed bright fluorescence under UV light were further tested. All the cultural and biochemical studies confirmed them to be P. fluorescens. The isolates also showed positive response for siderophore production and plant growth promoting activity on Chickpea cultivar Bital 98. Among these isolates Pf 3 and Pf 5 shown significant results by increasing root length and shoot length. Both the Pf 3 and Pf 5 isolates were found significantly superior than other isolates in increasing the shoot length (12.7 cm) and root length (24.5 cm) over control. The isolates Pf 3 was recorded high vigor index (3830) followed by Pf 5 (3648). The least vigor index was recorded by Pf 1 (2631).
Bio efficacy of pseudomonas fluorescens isolated from chickpea fields as plan...Shazia Shahzaman
Chickpea is an economically important food crop, which is subjected to infection by a host of fungal, viral and bacterial pathogens. Thirty isolates of Pseudomonas fluorescens were isolated from the rhizosphere of Chickpea fields. These were tested against F. oxysporum in dual culture method. Among these, four (Pf 1, Pf 3, Pf 5 and Pf
8) isolates were showed bright fluorescence under UV light were further tested. All the cultural and biochemical studies confirmed them to be P. fluorescens. The isolates also showed positive response for siderophore production and plant growth promoting activity on Chickpea cultivar Bital 98. Among these isolates Pf 3 and Pf 5 shown significant results by increasing root length and shoot length. Both the Pf 3 and Pf 5 isolates were found significantly superior than other isolates in increasing the shoot length (12.7 cm) and root length (24.5 cm) over control. The isolates Pf 3 was recorded high vigor index (3830) followed by Pf 5 (3648). The least vigor index was recorded by Pf 1 (2631).
Achievements and ongoing work on biopesticides at ICIPE—Some examples and les...ILRI
Presented by Jean Nguya K. Maniania, Sevgan Subramanian and Sunday Ekesi at the Regional Experts Workshop on Development, Regulation and Use of Bio-pesticides in East Africa, Nairobi, Kenya, 22–23 May 2014
Recent Advances in Biopesticides BY Ghulam Murtazamurtaza8513
Biopestides are being manufactured all across the world but due to limited resources the research in biopesticides is not upto the mark. however advancement has been made in recent decades to protect crops from the attack of different insect pest in order to meet the agricultural productivity.
Plant Growth-Promoting Activities and Molecular Characterization of Rhizobact...IOSR Journals
Rhizosphere bacteria are known to influence plant growth by direct and indirect mechanisms. Development of an effective plant growth promoting rhizobacteria (PGPR) inoculant necessitates the presence of a diverse set of traits that can help its colonization of the rhizosphere and survival under varying environmental conditions. In the present study, a total of 219 bacterial strains isolated from the rhizosphere of different medicinal and aromatic plants collected from different locations of Andhra Pradesh (India) were initially screened for their PGP activities. From the 219 isolates four bacterial strains were selected and tested for in vitro specific plant growth promotion activities such as ammonia production, IAA production, phosphate solubilization, HCN production and antifungal activity. These four isolates showed maximum plant growth promoting activities and further they were identified on the basis of colony morphology, gram staining and biochemical tests. These PGPR isolates were characterized through 16S rRNA gene sequencing which led to their identification as Pantoea sp. (Cf 7), Bacillus sp. (Cf 60) and Pseudomonas sp. (Te1, Av 30) respectively. Seed germination test was conducted by employing these strains under laboratory conditions on sorghum, maize and green gram seeds to investigate the effect of PGPR on the growth of seedlings. These PGPR isolates induced production of plant growth hormones (indole acetic acid), phosphate solubilization and ammonia production resulting in enhanced plant growth. Most of the isolates resulted in a significant increase in % of seed germination, shoot length, root length and vigor index of sorghum, maize and green gram seedlings. Therefore, the present study suggests that these PGPR isolates (Cf 7, Cf 60, Te1, Av 30) may be used as biofertilizers to enhance the growth and productivity of commercially important medicinal and aromatic plants.
Isolation, identification of antagonistic rhizobacterial strains obtained fro...Shazia Shahzaman
Plant growth promoting rhizobacteria (PGPR), are associated with roots, found in the rhizosphere and can directly or indirectly enhance the plant growth. In this study soil was collected from rhizosphere of chickpea fields of different areas of Rawalpindi division of Pakistan. PGPR were isolated, screened and characterized. Eight isolates of rhizobacteria (RHA, RPG, RFJ, RC, RTR, RT and RK) were isolated from Rawalpindi division and were characterized. The antagonistic activity of these PGPR isolates against root infecting fungi (Fusarium oxysporum and Verticillium spp.,) was done and production of indole acetic acid (IAA), siderophore and P-solubilization was evaluated. The isolates RHA, RPG, RFJ, RC, RRD and RT were found to be positive in producing siderophore, IAA and P-solubilization. Furthermore, most of the isolates showed antifungal activity against Fusarium oxysporum, and Verticillium spp. The rhizobacterial isolates RHA, RPG, RFJ, RC, RRD, RTR, RT and RK were used as bio-inoculants that might be beneficial for chickpea cultivation as the rhizobacterial isolates possessed the plant growth promoting characters i.e. siderophore, IAA production, phosphate solubilization. In in vitro tests, Pseudomonas sp. and Bacillus spp. inhibited the mycelial growth of the fungal root pathogens. The isolates (RHA and RPG) also significantly increased (60-70%) seed germination, shoot length, root length of the chickpea. The incidence of fungi was reduced by the colonization of RHA and RPG which enhanced the seedling vigor index and seed germination. The observations revealed that isolates RHA and RPG is quite effective to reduce the fungal root infection in greenhouse, and also increases seed yields significantly. These rhizobacterial isolates appear to be efficient yield increasing as well as effective biocontrol agent against fungal root pathogen.
Achievements and ongoing work on biopesticides at ICIPE—Some examples and les...ILRI
Presented by Jean Nguya K. Maniania, Sevgan Subramanian and Sunday Ekesi at the Regional Experts Workshop on Development, Regulation and Use of Bio-pesticides in East Africa, Nairobi, Kenya, 22–23 May 2014
Recent Advances in Biopesticides BY Ghulam Murtazamurtaza8513
Biopestides are being manufactured all across the world but due to limited resources the research in biopesticides is not upto the mark. however advancement has been made in recent decades to protect crops from the attack of different insect pest in order to meet the agricultural productivity.
Similar to Defence development of biopesticide for the control of root pathogenic fungi in chickpea using plant growth promoting rhizobacteria by shazia shahzaman
Plant Growth-Promoting Activities and Molecular Characterization of Rhizobact...IOSR Journals
Rhizosphere bacteria are known to influence plant growth by direct and indirect mechanisms. Development of an effective plant growth promoting rhizobacteria (PGPR) inoculant necessitates the presence of a diverse set of traits that can help its colonization of the rhizosphere and survival under varying environmental conditions. In the present study, a total of 219 bacterial strains isolated from the rhizosphere of different medicinal and aromatic plants collected from different locations of Andhra Pradesh (India) were initially screened for their PGP activities. From the 219 isolates four bacterial strains were selected and tested for in vitro specific plant growth promotion activities such as ammonia production, IAA production, phosphate solubilization, HCN production and antifungal activity. These four isolates showed maximum plant growth promoting activities and further they were identified on the basis of colony morphology, gram staining and biochemical tests. These PGPR isolates were characterized through 16S rRNA gene sequencing which led to their identification as Pantoea sp. (Cf 7), Bacillus sp. (Cf 60) and Pseudomonas sp. (Te1, Av 30) respectively. Seed germination test was conducted by employing these strains under laboratory conditions on sorghum, maize and green gram seeds to investigate the effect of PGPR on the growth of seedlings. These PGPR isolates induced production of plant growth hormones (indole acetic acid), phosphate solubilization and ammonia production resulting in enhanced plant growth. Most of the isolates resulted in a significant increase in % of seed germination, shoot length, root length and vigor index of sorghum, maize and green gram seedlings. Therefore, the present study suggests that these PGPR isolates (Cf 7, Cf 60, Te1, Av 30) may be used as biofertilizers to enhance the growth and productivity of commercially important medicinal and aromatic plants.
Isolation, identification of antagonistic rhizobacterial strains obtained fro...Shazia Shahzaman
Plant growth promoting rhizobacteria (PGPR), are associated with roots, found in the rhizosphere and can directly or indirectly enhance the plant growth. In this study soil was collected from rhizosphere of chickpea fields of different areas of Rawalpindi division of Pakistan. PGPR were isolated, screened and characterized. Eight isolates of rhizobacteria (RHA, RPG, RFJ, RC, RTR, RT and RK) were isolated from Rawalpindi division and were characterized. The antagonistic activity of these PGPR isolates against root infecting fungi (Fusarium oxysporum and Verticillium spp.,) was done and production of indole acetic acid (IAA), siderophore and P-solubilization was evaluated. The isolates RHA, RPG, RFJ, RC, RRD and RT were found to be positive in producing siderophore, IAA and P-solubilization. Furthermore, most of the isolates showed antifungal activity against Fusarium oxysporum, and Verticillium spp. The rhizobacterial isolates RHA, RPG, RFJ, RC, RRD, RTR, RT and RK were used as bio-inoculants that might be beneficial for chickpea cultivation as the rhizobacterial isolates possessed the plant growth promoting characters i.e. siderophore, IAA production, phosphate solubilization. In in vitro tests, Pseudomonas sp. and Bacillus spp. inhibited the mycelial growth of the fungal root pathogens. The isolates (RHA and RPG) also significantly increased (60-70%) seed germination, shoot length, root length of the chickpea. The incidence of fungi was reduced by the colonization of RHA and RPG which enhanced the seedling vigor index and seed germination. The observations revealed that isolates RHA and RPG is quite effective to reduce the fungal root infection in greenhouse, and also increases seed yields significantly. These rhizobacterial isolates appear to be efficient yield increasing as well as effective biocontrol agent against fungal root pathogen.
Distribution, Biochemical Properties and Genetic Relatedness of Endophytic Ba...AI Publications
Microbe-assisted phytoremediation is a recent application of bioremediation with much prospects. The genetic relatedness of culturable endophytic bacteria of wetland plants growing on a six month-old and twelve month-old petroleum-contaminated sites, and an uncontaminated site in Bayelsa State of the Niger Delta Region, Nigeria were compared. Most of the endophyte species isolated from the roots, stems and leaves were common to all the sites and belong to the phyla Proteobacteria, Bacteroidetes Firmicutes, Actinobacteria, Chloroflexi and Actinomicrobia, with the γ-Proteobacteria dominating. Pseudomonas was the most prevalent species in all three sites, but higher in the petroleum contaminated sites. Biochemical properties (API 20E) of the common isolates; Pseudomonas spp.Chryseobacterium indologenes,Bacillus and Proteusvaried with sites while only Providencia rettgeri peculiar to the petroleum-contaminated sites showed the same properties. 16S rRNA PCR-DNA fragments of forty-five species of the isolates (15 from each site) were characterized using RFLP and MspI restriction enzyme and a genetic distance tree of the restriction patterns drawn. The percentage of similarity in the genetic relatedness of isolates ranged from 11.1 – 100%. The genetic tree analysis of the 45 species of identified bacteria revealed 3 major clusters with 17 DNA fingerprinting patterns. Pseudomonas species of the root and leaves of the six month-old petroleum-contaminated site and uncontaminated site were seen to cluster together irrespective of date of isolation. The endophytes may play a role in the in situ degradation of the petroleum hydrocarbon of the sites.
Physiological Quality of Bean Seeds Related To Azotobacter spp. InoculationIOSRJAVS
Research aimed at improving the quality of crops. The results obtained with seed coating were very influential with regard to fertilization and disease resistance. Nitrogen fertilization increases costs in agricultural production and the loss of fertile lands, altering the natural conditions and has negative consequences for the microorganisms (MO), which regulate the balance between quality of crops and soils. The biological nitrogen fixation (BNF) consists on MO work which provide nitrogen in soil and secrete substances that promote plant growth, these can be made by inoculation of seeds. The germination rate, emergency and vigor are an indicator that determines the productivity and physiological quality of a plant variety. The germination rate, number of emerged seeds per pot in favorable field conditions and vigor by accelerated aging test were evaluated. The seed treatments were: the seed inoculation with Azotobacter spp., Immersion in nutrient broth without bacteria, sterilization and zero handling. The experiment has shown that inoculation of common bean seeds with Azotobacter spp. does not adversely affect germination, emergency or vigor and stimulate the development of abnormal seedlings.
Pseudomonas fluorescens as plant growth promoting Rhizo- Bacteria and biologi...Innspub Net
The use of plant growth promoting rhizobacteria (PGPR) to control disastrous diseases in many crops has been considered important recently. The research was conducted to evaluate several bacterial strains to control white rust in chrysanthemum. The research consisted of two chronological experiments, in vitro and in vivo testing of bacterial isolate against the disease. 16 bacteria isolates were collected, purified and applied on the rust-infected leaf. Three isolates showed more effective in suppressing white rust during in vitro testing and further identification confirmed these strains, Pf Kr 2, Pf Smd 2 and Pf Ktl were grouped into P. flourescens. In vivo testing of the Pf isolates also revealed consistent performances of these three Pf isolates in retarding the growth of fungal Puccinia horiana and even more effective than Azotobacter sp. and Azospirilium sp. The production of ethylene on the leaf was coincidence with the slower development and lower disease intensity on the treated plants. Among the three strains, Pf Kr 2 showed stronger suppression to the disease. Further investigations are needed to further elucidate the existence of specific interrelation between Pf strains and plant genotypes or cultivars. Prior to a selection of good bacterial inoculants, it is recommended to select cultivars that benefit from association with these bacteria. Get the full articles at: http://www.innspub.net/ijaar/pseudomonas-fluorescens-as-plant-growth-promoting-rhizo-bacteria-and-biological-control-agents-for-white-rust-disease-in-chrysanthemum/
Abstract— This study was conducted to identify, test the pathogenicity of strawberry root and stalk rot pathogens and evaluate the efficiency of some biocontrol agents and fungicides to control the disease. The isolation and identification of fungi associated with infected plant samples showed that Rhizoctonia solani was detected in all studied commercial strawberry lath houses at different location of Baghdad-Iraq. The frequency percentages ranged 25.5-63.5 % and 10.75 - 40 % for Rhizoctonia solani and Phymatotrichopsis omnivora respectively. Pathogenicity test revealed R. solani and P. omnivora isolates were highly pathogenic to strawberry plants. The disease severity percentages of R. solani and P. omnivora were 83.0-100% and 55.5-62.0 % respectively. The isolates HRs3 and KPh1 of R. solani and P. omnivora respectively, caused the highest disease were used during this study. The control agents Rizolex and Tachigarin fungicides, Azotobacter chroococcum and Pseudomonas fluorescens have shown high efficiency against R. solani and P. omnivora on culture media (PDA).
The treatment of biocontrol agent’s A. chroococcum and P. fluorescens and the fungicide Rizolex and Preserve Pro showed high efficiency in disease control and enhance plants growth under greenhouse conditions. Disease severities on foliar and root system in A. chroococcum , Rizolex , Preserve Pro and P. fluorescens were 6,66 and 0.00 %, 20.00 and 0.00 %,13.33 and 0.00 % and 13.33and 0.00 % respectively in plants infected with R. solani .Whereas they were 6.66 and 0.00%, 13.33 and 0.00 %,13.33 and 0.00 %,and 13.33 and 0.00 % respectively in plants infected with P. omnivora. This study is the first report of the occurrence of root and stalk rot disease caused by R. solani and P. omnivora on strawberry plants in Iraq.
Pesticidal efficacy of crude aqueous extracts of Tephrosia vogelii L., Allium...researchagriculture
Cabbage aphid (Brevicoryne brassicae L.) is one of the most problematic pests in smallholder vegetable production, causing significant yield losses in heavy infestations. Current control strategy focuses on use of synthetic pesticides that consequently lead to decimation of natural enemies, development of insect resistance and resurgence and upset biodiversity. Botanical pesticides have been used widely in smallholder farmers but not much documented literature exists on efficacy of these products. A field trial was done to assess the efficacy of crude aqueous extracts of Tephrosia vogelii, Allium sativum and Solanum incanum in controlling Brevicoryne brassicae in Brassica napus production. The trial was laid in a randomized complete block design (RCBD) with five treatments replicated four times. The five treatments used in the experiment were T. vogelii, A. sativum, S. incanum, dimethoate and control. Wingless adult female aphids were inoculated three weeks after transplanting of seedlings. Spraying and data collection were done weekly for four weeks. Data was collected on aphid nymph and adult counts on the third leaf from the aerial plant part of randomly selected plants from each treatment for 24 hours after the application of treatments and total plant fresh weight per each treatment. There were significant differences (p<0.05)><0.05) on the yield of rape. It was concluded that T. vogelii, S. incanum and A. sativum aqueous crude extracts have some pesticidal effects on aphid in rape production.
Article Citation:
Shepherd Mudzingwa, Simbarashe Muzemu and James Chitamba.
Pesticidal efficacy of crude aqueous extracts of Tephrosia vogelii L., Allium sativum L. and Solanum incanum L. in controlling aphids (Brevicoryne brassicae L.) in rape (Brassica napus L.)
Journal of Research in Agriculture (2013) 2(1): 157-163.
Full Text:
http://www.jagri.info/documents/AG0040.pdf
Pesticidal efficacy of crude aqueous extracts of Tephrosia vogelii L., Alli...researchagriculture
Cabbage aphid (
Brevicoryne brassicae
L.) is one of the most problematic
pests in smallholder vegetable production, causing significant yield losses in heavy
infestations. Current control strategy focuses on use of synthetic pesticides that
consequently lead to decimation of natural enemies, development of insect
resistance and resurgence and upset biodiversity. Botanical pesticides have been used
widely in smallholder farmers but not much documented literature exists on efficacy
of these products. A field trial was done to assess the efficacy of crude aqueous
extracts of
Tephrosia vogelii
,
Allium sativum
and
Solanum incanum
in controlling
Brevicoryne brassicae
in
Brassica napus
production. The trial was laid in a randomized
complete block design (RCBD) with five treatments replicated four times. The five
treatments used in the experiment were
T
.
vogelii
,
A
.
sativum
,
S
.
incanum
,
dimethoate and control. Wingless adult female aphids were inoculated three weeks
after transplanting of seedlings. Spraying and data collection were done weekly for
four weeks. Data was collected on aphid nymph and adult counts on the third leaf
from the aerial plant part of randomly selected plants from each treatment for
24 hours after the application of treatments and total plant fresh weight per each
treatment. There were significant differences (p<0.05)><0.05) on the yield of rape. It was concluded that
T. vogelii
,
S
.
incanum
and
A
.
sativum
aqueous crude extracts have some pesticidal
effects on aphid in rape
production.
Determination of antibacterial activity of various broad spectrum antibiotics...Open Access Research Paper
Bacterial leaf blight (BLB) of rice (Oryza sativa L.) caused by Xanthomonas oryzae pv. oryzae, is arguably the most holistic pathosystem of rice throughout the worldwide due to its growing concern as this disease is wide spread, devastating and its control measures are still not well understood. In vitro evaluation of various broad spectrum antibiotics viz., streptomycin sulphate, kanamycin sulphate, chloramphenicol, ampicilin trihydrate and benzylpenicillin, was carried out to determine the best chemistry against the destructive pathogen Xanthomonas oryzae pv. oryzae at different concentrations. Inhibition zones appeared on petri plates for the growth of bacteria were very clear around the paper disks. Chloramphenicol proved to be the most effective antibiotic to control the bacterium as it suppressed the bacterial growth to greater extent and only the 6.25 mean bacterial colonies were appeared in the petri plates, followed by the ampicillin trihydrate which showed to be the second most effective antibiotic against the pathogen growth and retarded to 12.00 mean bacterial colonies. The maximum diameter of inhibition zone (28.31 mm) was showed by the Chloramphenicol at 100 ppm followed by ampicillin trihydrate which gave proved to be second most effective antibiotic to control the pathogen and gave maximum inhibition zone (25.02 mm) at 100 ppm concentration. All the antibiotics showed significant results at higher concentrations. The study suggests that the experiments in the field must be conducted to prove the effectiveness of these broad spectrum antibiotics in the natural environmental conditions as there is a possibility of some variation in the field results because of various factors which influence the chemical management of plant diseases in the field.
Biofumigation: A Potential Aspect for Suppression of Plant-Parasitic NematodesIJEABJ
Plant-parasitic nematode cause economic loss to crops throughout the world. Biofumigation is the environmental friendly control option for the suppression of plant-parasitic as well as other pathogenic soil microbes. Glucosinolates are the main active compound present in some plants which are responsible for biofumigation process. To increase the efficiency of biofumigation selection of varieties containing more glucosinolates is highly desirable. Plant growth stage, soil temperature, soil texture, moisture, soil depth and soil microbes play important role in efficient biofumigation.
Mass Production of Paecilomyces Lilacinus by using Different Cultivation Medi...Agriculture Journal IJOEAR
Paecilomyces lilacinus is a common saprophytic, filamentous fungus. Morphological characters of Paecilomyces lilacinus were separate mycelium, hyaline, conidia white to pink colored and formation of phialides. The growth of Paecilomyces lilacinus carried out on SDA media at room temperature was better than incubator. Various solid substrates like Rice, Wheat bran, and Sorghum were evaluated for the mass multiplication of fungus Paecilomyces lilacinus. Added dextrose and antibiotics in solid media for mass multiplication at room temperature. Among all the substrate Wheat bran recorded the maximum spore count of 7. 1 10-8 spore/ml followed by Sorghum 5. 4 10-8 spore/ml and Rice 5. 1 10-8 spore/ml after 20 days. Also dry mycelia weight or biomass of fungus Paecilomyces lilacinus without an incubator was more than using an incubator.
Prevalence, occurrence and biochemical characterization of Xanthomonas campes...INNS PUBNET
Xanthomonas campestris pv. vesicatoria the causal organism of bacterial spot in tomato results in heavy losses both in the form of quality and. In this study a survey was carried out to report the incidence of bacterial spot disease of tomato in district Swat. We reported maximum disease incidence in tehsil Kabal (71.66%), followed by Charbagh (61.66%) and Barikot (58.33%). For resistant screening a total of 13 tomato germplasms were screened against the disease. The foliar severity ranged from 3.33% to 73.33%, while severity for fruits was ranged from 18.33% to 30.66%. In case of phenotypic data the highest numbers of fruits obtained were 34, plant height 79.5cm and fruit weight was 470 grams/ten tomatoes. While the lowest average numbers of fruits were 6.67, plant height 45.7cm and fruit weight recorded was 215.67 grams/ten tomatoes. Line 1288 showed highest level of resistance followed by Red-stone. However, line 9708 showed highest susceptibility when exposed to artificial inoculation. Our study showed that bacterial spot is a major issue in some part of Pakistan and germplasm screening are linked to increased host resistance and could offer an important contribution to future integrated bacterial spot management programs.
The study was carried out with the aim of sourcing for bacteria from the natural environment having antifungal capabilities to control and inhibit postharvest fungal spoilage of fruits and vegetables caused by Botrytis cinerea. Soil and water samples were collected from Heriot Watt University environment and Dr Ruth Fowler’s garden and inoculated using the spread plate technique; identification was carried out using Microbact Identification kits; and isolates assayed for antifungal activities against Botrytis cinerea. Forty eight bacteria species were isolated out of which sixteen (16) belonging to genera Pseudomonas, Bacillus, Escherichia, Burkholderia, Staphylococcus, Streptococcus, and Proteus showed antifungal activities. Bacteria species Pseudomonas stutzeri and Burkholderia cepacia had the highest zones of inhibition with average radii of 3.06 and 3.20 cm respectively. The bacteria had the potential to inhibit mycelial and spore growth at varying levels thus making them possible candidates for further tests and studies. Considering the aim of the study, further research into identifying these antifungal isolates inhibitory compounds and metabolites is highly recommended.
Nodulation, Growth and Yield Response of Five Cowpea (Vigna unguiculata L. Wa...Premier Publishers
The experiment was carried out in the screen house of the Department of Crop, Soil & Pest Management, Federal University of Technology, Akure, Nigeria. The experimental layout was a 5 x 3 x 2 factorial combination with 3 replications given a total of 90 treatments. Seeds of five cowpea varieties namely: IT98K-205-8, Ife Brown, Oloyin Brown, IT98K-573-2-1 and IT96D-610 were sown in Plastic buckets of 7-liter capacity and were perforated at the bottom to allow for drainage and filled with top soil. Watering regimes of (500ml, 700ml and 900ml) were imposed and water was applied twice a week while cowpea plants were inoculated with 5g each of Rhizobia strain (Mesorhizobia loti) at seedling stage. Control set was maintained without inoculation. The effect of watering regimes on legume species was significant on nodulation, growth and yield characters of cowpea varieties evaluated. The results revealed marked varietal differences in plant growth, nodulation, yield and yield components. IT98K-573-2-1 and Oloyin Brown generally expressed superior performance in most measured parameters. Mesorhizobia inoculation significantly (p≤0.05) increased plant growth, nodulation, yield and yield components of cowpea. The interaction effect of variety, Mesorhizobia loti and watering regimes caused significant variations in the number of nodules, leaf area, number of seeds/pod and seed yield. The nitrogen and crude protein content in the leaf differed among the cowpea varieties evaluated. Application of mesorhizobium strain significantly increased seed yield of cowpea and caused substantial increase in nodulation and this subsequently affected the Nitrogen fixation potential of cowpea under varying soil moisture regimes.
CHARACTERIZATION OF STREPTOMYCES SCABIES ISOLATESijabjournal
Potato, (Solanum tuberosum L,) have various biotic constraints in its production due to pest attack. Among these, common scab caused by streptomyces scabies in an important disease in potato which causes economic loss with respect to plant yield and quality of tubers. The present study was conducted to determine the pathogenicity, pathogenic variation, characterization of morphological, physiological and
biochemical aspects of Streptomyces specie associated with potato tubers grown in Rawalpindi district.Severity data and pathogenic variation of disease was studied by using different isolation and characterization techniques. Isolation and characterization of Streptomyces spp. From potato tubers will
guide the researchers about the causative strains of common scab of potato present in the particular area.
Similar to Defence development of biopesticide for the control of root pathogenic fungi in chickpea using plant growth promoting rhizobacteria by shazia shahzaman (20)
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
This pdf is about the Schizophrenia.
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Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
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Defence development of biopesticide for the control of root pathogenic fungi in chickpea using plant growth promoting rhizobacteria by shazia shahzaman
3. Prof. Dr. M. Inam-ul-Haq
Department of Plant Pathology
Prof. Dr. Tariq Mukhtar
Department of Plant Pathology
Prof. Dr. M. Naeem
Department of Entomology
Supervisor
Member
Member
4. INTRODUCTION TO CROP
Chickpea (Cicer arietinum L.) belongs to Fabaceae
family and ranked third after dry beans and peas
(FAO, 2013)
Significant in Human diet and animal feed.
Chickpea cultivated area is 1068 thousand ha-1 with
production of 523 thousand tons during 2013-14 with
an average yield of 685 kg ha-1
Agri. Stat. of Pak., 2014
5. PAKISTAN RANKING IN CHICKPEA PRODUCTION
77%
9%
7%
4% 3%
India
Turkey
Pakistan
Islamic republic of Iran
Mexico
Pakistan ranked 3rd in world chickpea
production 757.1 (Kg/Ha).
WORLD PRODUCTION
Data Source
http://www.factfish.com/statistic country/Pakistan/chickpea,+ production+ quantity
The Punjab province alone contributed 900.1 thousand
ha which was 84% of the to the total chickpea area
grown in the country.
6. Area and Production of Chickpea Crop
Crop
2012-2013 2013-2014
% change in
ProductionArea
(000 ha)
Production
(000 tons)
Area
(000 ha)
Production
(000 tons)
Chickpea 1067 562 1068 523 -6.9
670
680
690
700
710
2012-13 2013-14
Yield (kg/ha)
7. About 10-50% losses by Fungal Root Disease
have been reported on chickpea in the dry areas of
Pakistan during the past several years
Woltz and Jones, 2012
Estimated yield losses in Chickpea due to
biotic and abiotic factors range from 15-80%
Kuku et al., 1996
8. Wet Root rot Fusarium Wilt
Verticillium Wilt
Important Fungal Diseases of Chickpea
Dry Root rot
9. Need of the Project
• 311 compounds have been registered as
fungicides Milne, 2010
• Fungicides results into acute and chronic toxicity.
Goldman, 2008
• Environmental pollution result in human
exposure through consumption of residues of
pesticides in food and, possibly, drinking water
Suprapta, 2012
• Millions of people suffer from pesticide
problems and 18,000 die every year. WHO/
Miller, 2012
10. • Various reports on the utilization of potential
microbes as bio-control.
Yang et al., 2008
• Several species of Rhizobacteria antagonize
the fungal pathogens.
Walsh et al., 2001
• Development of Bio-formulation using Plant
growth promoting rhizobacteria (PGPR).
Kumar et al., 2011
12. Hypothesis
Rhizobacteria naturally present in soils may interfere with the
extent of root colonization, Disease suppression and plant growth
promotion by Plant Growth Promoting Rhizobacteria (PGPR)
13. • Isolation of fungal root pathogens and rhizobacteria from chickpea
roots and rhizospheric soil.
• Selection of rhizobacterial isolates antagonistic to root pathogenic
fungi of chickpea and their characterization.
• Evaluation of PGPR based formulations against fungal root pathogens
under various conditions.
• Selection of suitable formulation as a biopesticide.
14.
15. Plant growth promoting rhizobacteria (PGPR) were first defined as
the soil bacteria that colonize the roots of plants by following
inoculation on to seed and that enhance plant growth
Kloepper and Schroth
(1978)
At present, the use of biological approaches is becoming more popular
as an additive to chemical fertilizers for improving crop yield in an
integrated plant nutrient management system. In this regard, the use of
PGPR has found a potential role in developing sustainable systems in
crop production.
(Sturz et al. 2000;
Shoebitz et al.2009).
P. fluorescens strains isolated from rhizosphere of rice, wheat, pigeon
pea, groundnut and chili crops produced extra cellular siderophores
which were antagonistic to fungal pathogens like Fusarium
oxysporum, Alternaria sp and Colletotrichum capsicii.
(Suryakala et al.,2004)
Important genera of bacteria used in natural and man-created
bioremediation includes Bacillus, Pseudomonads, Methanobacteria,
Ralstonia and Deinococcus, etc.
Milton 2007
The production of siderophores with fungicidal properties of
Pseudomonas fluorescens and neem cake against Fusarium wilt
disease of chickpea. The potential them is worthy of further evaluation
as a biocontrol system for chickpea wilt.
Inam et al., 2010
16. Utilization of these organisms has no negative effect on
environment and also on other non-target organisms.
Various reports are available on the utilization of these
potential microbes as bio-control agents. Yang et al., 2008
Several soil borne non-pathogenic species of rhizobacteria
are reported to antagonize the disease causing fungal
pathogens and can be utilized as alternative to chemical
control measure Walsh et al., 2001
PGPR belonging to various genera, such as Azotobacter,
Azospirillum, Pseudomonas, Acetobacter, Burkholderia and
Bacillus and have been repeatedly reported by many
researchers. Bashan and Ulangatan, 2002
Many bio-control based formulations have been prepared
and commercialized in America and Europe. Many of the
pesticide companies aimed to develop bio-control gents
based bio-pesticide formulations as commercial product.
Suprapta, 2012
Increase in soil fertility, plant growth promotion, plant
pathogen suppression and development of eco-friendly
bio-formulation is the matter of major concern. Arora et al., 2010
19. An extensive survey of Rawalpindi, Chakwal and
Attock District was done in 2012-13
20. Rhizospheric soil and root sample collection
Random fields in each visited Districts
Random samples from each visited field
Hierarchical sampling strategy was followed for sampling
(McDonald and Martinez, 1990)
25. Isolation of Fungal Root Pathogen
• Infected root tissue at the advancing edge of a wilted area and
rhizosphere soil were used.
• Tissues were washed and cut into small pieces (2 mm), surface
disinfested in 1 % NaClO for 2 min, rinsed in distilled water,
and damp-dried on absorbent paper towels before being plated
on the PDA media.
(Jose et al , 2012)
26. Fig.2 Characterization of fungal root pathogen on the basis of colony shape and color A. Rhizoctonia
spp. B. Pythium spp. C. Fusarium spp. E. Macrophomina spp.
a b c
d e f
27. Fig. 3: Microscopic and visual observation of Fungal Root Pathogens
A- Colony growth of Fungal Root Pathogens
i.e., Fusarium spp., Rhizoctonia spp., Macrophomina spp.
28. Fig.4 Observation under microscope at 40 X (a) Rhizoctonia spp. (b) Fusarium spp. (c)
Fusarium spp. (d) Macrophomina spp. (e) Pythium spp.
a
b
c d e
a
d e
29. Morphological Identification of Fungal Root Pathogens
Fungal Isolates Colony Color Spore Shape Macroconidia
(µm)
Colony
Diameter(mm)
Media Used
Fusarium spp. Brick red to
violet
Blunt 21.49-29.23 3.2 Potato
Dextrose Agar
Rhizoctonia
spp.
Brownish to
blackish
Papillate 12.3-15.0 5 Potato
Dextrose Agar
Pythium spp. Slightly yellow Tapering 10.5-12.0 2.9 Corn Meal
Agar
Macrophomina
spp.
Creamy to pink Hooked 18.3-20.5 3.5 Potato
Dextrose Agar
(Arotupin, 2004)
30. Fungal Root Pathogen Isolated
Tehsil Fusarium
spp.
Rhizoctonia
spp
Pythium
spp.
Macrophomina
spp.
Total
FatehJang 9 3 5 4 21
Hasanabdal 9 3 3 3 18
Pindigheb 5 4 3 2 14
Chakwal 10 7 4 5 26
Doltala 6 5 3 2 16
Tarnol 5 4 3 2 14
Taxila 5 1 1 1 8
Kahuta 6 2 1 2 11
Total 55 29 23 21 128
Table.2 Total 128 fungal root pathogens isolated from rhizospheric soil and roots and most
of them from Chakwal i.e. 24
31. Isolation of Rhizobacteria
• Soil samples 1 g of each sample was suspended in 9 ml sterile
water and shaken vigorously for 2 min.
• The soil suspension was serially diluted (from 10-1 to 10-9)
• 0.1 ml of all dilutions were plated on selective media
supplemented with a commercial antifungal to inhibit fungi
growth.
• Petri dishes were incubated at 28±2°C
(Gerhardt, 1994)
32. Morphological Characterization of Rhizobacteria
Bacterial
Isolates
Colony
shape
Colony
Margin
Colony
Elevation
Colony
Color
Gram
staining
Media used
RB1 irregular flat flat white -ve Azotobacter
Agar
(Mannitol)
RB2 irregular round flat Creamy
white
+ve Nutrient Agar
RB3 irregular oval raised Yellowish
white
-ve King B
RB4 slightly
dome
shaped
round raised yellow -ve King B
RB5 Circular pointed raised yellow -ve Tryptic soy
agar
36. • Inoculum of isolated fungal root pathogens was mixed with
1kg autoclaved soil in pots.
• Five seeds of chickpea cultivar were sown in each pot and
grown for 40 days at 25±2°C.
• Control plants were grown in a comparable mixture of
uninfected and autoclaved soil.
37. • Typical wilt symptoms were observed in most of the pots in 41
days after sowing, development of symptoms were recorded.
• Re-isolation of fungal root pathogens was made by taking
infected plant tissues on selective media, Identification of
these pathogens indicating that the plant mortality was
associated with these fungal root pathogens.
Nene & Haware (1980)
48. (Dual Culture Technique)
• Antagonism was tested by culturing both rhizobacteria and
Fungal Root Pathogens on same culture media containing plate.
• Dual culturing of rhizobacteria and pathogenic fungi
Incubation at 26±2oC.
• Control contain only pathogen
• Data collection by measuring zone of inhibition in mm.
Haine et al. 2008
49. (Poisoned Food Technique)
• The systemic fungicides viz., Tilt, Contaff, Bavistin and Folicur
were evaluated against the test fungus at the concentration of 15
ppm.
• Each media toxicated with fungicide was poured in three Petri plates
• Non toxicated media was poured into Petri plates kept as a check.
• A 5 mm mycelia disc of 6 days old culture of the test pathogen was
cut with sterile cork borer and placed in center of each Petri plate.
(Sharvelle, 1995)
59. • A field trial was conducted with two local varieties of
Chickpea Bital-98, Desi-sp, in already infested field at
FatehJang in a randomized block design with three
replications.
• Plot size use was 2×2 m2. Chickpea seeds were treated with
rhizobacterial isolates (seed bacterization) following Jayraj et
al. (1999).
• The number of rhizobacterial population was maintained
through serial dilution and plating in NA. Treatments consist
of:
60. T1: RB-1(108 cfu/ml) @ 50ml/kg of seed
T2: RB-2(108 cfu/ml) @ 50ml/kg of seed
T3: RB-3(108 cfu/ml) @ 50ml/kg of seed
T4: RB-4(108 cfu/ml) @ 50ml/kg of seed
T5: RB-5(108 cfu/ml) @ 50ml/kg of seed
T6: control (no treatment)
61. Statistical Analysis
Data obtained from in vivo test were pooled for statistical
analysis and subjected to one ways ANOVA for determining any
significant differences among the treatments.
62. Table. 3 Effect of different Rhizobacteria on various attributes in Chickpea varieties cultivated
in wilt affected field after 60 days V1: Bital-98, V2: Desi-sp; variety; data are mean of three
replications
Treatments Plant Height (cm) # of nodules/plant Dry weight (g/plant)
V1 V2 V1 V2 V1 V2
T1 32.2 60.8 4.66 13.00 1.8 3.0
T2 26.8 80.0 10.33 13.66 1.0 3.2
T3 31.0 71.5 7.33 11.00 2.1 3.5
T4 30.2 48.4 14.00 16.00 2.3 2.8
T5 37.2 91.2 6.00 12.66 2.7 6.5
T6 26.7 24.1 1.60 2.00 0.5 1.4
CD (p=0.05) 7.21 3.31 1.86
63. Table.4 Effect of various Rhizobacteria on disease parameters of Fungal Root disease in Chickpea
average value of occurrence of disease for three months
Treatments Disease Incidence (%) Disease Severity (%) Percentage Disease
Control
T1 52.67 53.11 35.03
T2 61.67 60.91 30.30
T3 50.67 54.08 34.48
T4 48.00 51.00 41.59
T5 47.00 49.37 43.63
Control 97.00 96.25
CD (p=0.05) 11.13 10.04
14 isolates of rhizobacteria were selected on the basis of their antagonism against
Fungal Root Pathogens i.e. 6 of T4 and 8 from T5
65. Biochemical Test Performed
• Gram’s reaction
• Carbohydrate fermentation
• Oxidase test
• H2S production
• Catalase activity
• Casein hydrolysis
• Indole production
• Urease test
• Acid and gas production
• Starch and gelatin hydrolysis
(Cappuccino and Sherman, 1992)
67. Fig.6 Biochemical test for Rhizobacterial isolates i.e, a. Catalase activity c. KOH test d.
Gram staining e. Starch hydrolysis f. Gelatin liquefaction g. Indole production
a b c d
e f g
d
h
70. PRIMER USED FOR RHIZOBACTERIA
Primers designed based on already published universal primers (e.g. 16S rDNA)
were used
.
Van der Meer et al., 2010
71. PRIMER USED FOR FUNGAL ROOT PATHOGNS
Primers designed based on already published universal primers (e.g. 16S rDNA)
were used
.
Aoki T, et al., 2003
72. DNA Extraction
• Extraction buffer that contains
detergent cetyl-tri-methyl ammonium
bromide (CTAB) and 2-β-
mercaptoethanol, EDTA and polyvinyl
pyrolidone (PVP).
• Quantification of DNA was done
with spectrophotometer determination.
Working concentration of DNA was
adjusted to 20 ng/ml and stored at 4ºC
Martins et al., 2005
73. PCR (RHIZOBACTERIA)
The PCR thermal profile consisted of an initial denaturation step
(94°C, 3 min), followed by 30 cycles at 94°C (30 sec), 55°C (1
min), 72°C (90 sec) and a final elongation step of 10 min at 72°C.
Kuske et al. ,1997
74. • Five µ l of the amplification products (amplicons) were
analysed on 1% (w/v) agarose gels cast and run in TAE buffer
(0.04 M Tris, 0.001 M EDTA, 0.02 M acetic acid), stained with
ethidium bromide and photographed under UV translluminator.
M RB-1RB-2RB-3 RB-4 RB-5RB-6RB-7 RB-8 RB-9 RB-10RB-11
1 2 3 4 5 6 7 8 9 101 1211 13
M
75. PCR (FUNGAL ROOT PATHOGEN)
Reactions involved 1 cycle at 95°C for 5 min, followed by 35 cycles with a
denaturation step at 95°C for 30 s, an annealing step at 55°C for 1 min, and an
extension step at 72°C for 1 min, followed by 1 cycle at 72°C for 6 mins
F1 F2 F3 F4 F5 F6
1 2 3 4 5 6M
76.
77. SEQUENCING AND ANALYSIS
• Sequencing (MACROGEN)
• Basic Local Alignment Standard Tools (BLAST)
• Nucleospin Extract Kit (Macherey Nagel, Germany) and
sequenced at Genelab Casaccia (S. M. Di Galeria, Italy).
MEGA software used for phylogenetic analysis to check
similarities.(Zheng et al.,2000), using default parameter
values, to give the percentage homology with known
sequences in the NCBI database.
78. Fungal Root Pathogen Identified
Isolates Accession number
Fusarium oxysporum EU091063
Macrophomina Phaseolina EU091058
F. oxysporum GU126793
F. oxysporum EU091056
F. oxysporum JF740777
Rhizoctonia solani DQ8376901
Fusarium sp. KJ776745
90. • Organic manure (O.M), vermi compost (V) Rice bran (Rb),
wheat bran (W) and decomposed mustard oil cake (D) were
collected from the local markets.
• Carboxymethyl cellulose (1% aq) was used as adhesives.
• The pH was adjusted to 7
• The mixture was then spread in a sterilized non sticky
disposable plate under sterile conditions Mannitol was added
as osmoticant.
91. • Subsequently, antagonistic rhizobacterial cell suspension of
concentration of 108 cfu/ml was pipetted into the mixture (1:10
v/w) and thoroughly mixed with the help of sterilized spoon.
• Formulation was divided into 3 parts, packed separately in
polypropylene bags (8 x 6.5 cm), heat sealed and stored at
28ºC temperature.
• Another set of bio-formulations was prepared for each
substrate carrier-adhesive bio-agent mixture and stored at 5°C
for comparative study.
Bora and Deka (2007)
92.
93. The population dynamics of the bioagents was determined
at different days after storage (DAS) in the two storage
conditions.
At 7, 30, 60, 90 and 120 DAS at 5◦C and 28◦C temperature,
Population dynamics was examined by mixing 1 g of
formulations aseptically with 10 ml sterile distilled water
for 20 min in a rotary shaker.
After incubating the plates at 28 ± 1°C for 48 h, the cfu/g
formulations were counted out. The population of Bioagents
in powder formulations (cfu/g formulation) recorded were
transformed and used for analysis in this study.
96. Effect of various carrier based formulations of selected antagonistic rhizobacterial strains on 7th day after
storage. The growth responses were analysed at the end of 7th day.
0
10
20
30
40
50
60
70
80
90
100
5◦C 28◦C
98. Effect of various carrier based formulations of selected antagonistic rhizobacterial strains on 30th day after storage. The growth
responses were analysed at the end of 30th day.
0
50
100
150
200
250
300
350
400
450
500
5◦C 28◦C
100. Effect of various carrier based formulations of selected antagonistic rhizobacterial strains on 60th day after
storage. The growth responses were analysed at the end of 60th day.
0
100
200
300
400
500
600
700
800
900
1000
5◦C 28◦C
102. Effect of various carrier based formulations of selected antagonistic rhizobacterial strains on 90th day after storage.
The growth responses were analysed at the end of 90th day.
0
10
20
30
40
50
60
70
80
90
100
5◦C 28◦C
104. Effect of various carrier based formulations of selected antagonistic rhizobacterial strains on 120th day after
storage. The growth responses were analysed at the end of 120th day.
0
10
20
30
40
50
60
70
80
90
100
5◦C 28◦C
105. Somasegran and Hoben (1994)
Fig.6 Colony count after different days after storage to check efficacy of selected antagonistic rhizobacterial strains
108. POT EXPERIMENT
• Surface sterilized chickpea seeds with disinfested in 1 % NaClO for
2 min, rinsed in distilled water, and damp-dried on absorbent paper
towels before sowing
• Seed bacterization 5g/kg of seeds
• Plants were inoculated with pathogen at 36 days after sowing
• Disease intensity was assessed 20 days after inoculation
• Control pots with only infested soil.
• All experiments were repeated with three replicates of each
treatment. Experiments were carried out in green house in a
complete randomized design (CRD).
112. • Seeds were surface-sterilized with 0.1% sodium hypochlorite .Later,
50 g of chickpea seeds were treated with 0.5 g of bio-inoculant
carrier formulations.
• Chickpea variety Bital-98 was sown with 4 replications, 4
treatments in 4 selected fields.
• The controls included only the pathogen inoculated treatment with
no bio-formulation applied (control)
• The experiment was laid out in a randomized complete block design
with factorial arrangement. The net plot size for the experiment was
1.6 x 5 m.
• Seed was drilled in 40-cm spaced rows and Plant-to-Plant distance
of 15 cm was maintained by thinning ten days after germination in
experiments.
120. Growth Parameters Analyzed
Treatment Germination shoot length (cm) Root length Fresh weight (g) Dry weight (g)
Control 53.75c 16.75e 3.92e 0.47e 0.075e
Bs-1OM 61.25b 24.1a 5.47c 0.73bc 0.16c
Bs-2OM 73.25b 28.55b 9.25ab 1.36a 0.37b
BS-3OM 89.25a 32.75a 12a 1.61a 0.51a
PfOM 69ab 26.4b 7.425bc 0.92c 0.24b
The growth responses were analysed at the end of 100 days. The values represent the mean
of 20 replicates (±SD). Means followed by the same letter in a column are not significantly
different from each other according to Duncan’s multiple range test.
121. Conclusion
• Selection of different antagonistic rhizobacteria by using
various cheap carrier materials.
• Bacillus subtilis (Bs-3) used with Organic Matter (O.M)
showed best results in Lab, green house as well as in Field
experiments among all.
• So, we recommend selected bio-formulation for the control of
fungal root pathogens to get the best results with maximum
yield.
• As this bio control is very cheap and environment friendly, so
it will help the poor farming community
122. Expected Outcomes
They are generally less destructive to Beneficial's, cause less
environmental pollution than conventional pesticides.
Biopesticide may provide a satisfactory alternative to chemical
pesticides when used as part of an overall IPM plan.
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Editor's Notes
most important source of vegetable protein. “hence they are quite rightly called poor man’s meet”
Among major chickpea producing countries Pakistan occupies 3rd position and India is top leading producer while within Pakistan, Punjab is major province with 87% production
Several factors are involved in lowering the yield of crop
In order to cope with these noxious pathogens farmer have to apply synthetic agro chemicals on intensive scale, which are not ecofriendly and also toxic to other non-target organisms it has been repotted that 311….due to application of these fungicides millions of people die every year
There is a need to find some alternate soln to replace these synthetic chemicals ………effectiveness of various microbes belong to microbial group are available….among these rhizobacteria have been reported to antagonize fungal pathogens in a very effective way due to their efficient weaponry system
Direct effect is nitrogen fixation, production of plant hormones, enhanced iron availability, phosphorus solubalization
Indirect effect antibiosis, induced resistance, iron scavenging, competing for nutrients and niche, parasitism and predation
In order to meet our objectives following methodology was used
seedling (October-November) and reproductive stage (February-March) random places across a diagonal in each of the selected field
This adopted procedure reduced the chance of bacterial contamination.
Following morphological characters were used to identify Fungal isolates
As the max disease severity observed in FatehJang so, max pathogenic isolates of following FRP were collected from these areas including fusarium rhizoctonia….and totla 128 isolates……
Pathogenicity test was performed to check virulence of isolated FRP
On the basis of disease severity rating scale pathogenicity was confirmed
Following isolates were tested for their virulence on these 5 susceptible chickpea cultivars….+++ indicates high virulence….
Colonies with more common type of morphology were selected
RB isolates were screened out for their antagonistic activity against selected most virulent FRP i.e., 80
Antagonist and FRP counter grown on same medium (PDA) plates 3 replications under CRD design
Among the tested RB isolates RB2 resulted in max zone inhibition of Fusarium spp. Followed by RB4 and so on
a field trail was conducted for screening of RB isolates wd 2 local varieties
Rhizobacteria cell suspension was prepared by culturing in conical flask containing Nutrient broth and kept at incubator for three days at 28±1ºC
Following treatments were made and applied
Data regarding disease severity and growth parameter was checked by comparing with control experiment
Most suitable antagonistic isolates were selected for further characterization
Further confirmation was done to check similarities and genera of isolates
16S r DNA was amplified by using thses two primer sets
Total genomic DNA isolation was confirmed by running on 1% agarose gel
PCR was done by using template DNA
Reaction were carried out in a 50 μl reaction mixture containing 0.2 mM dNTPs, 2 mM MgCl2, 0.5 μ M of each primer, 2.5 units of Taq DNA polymerase and 1x PCR buffer