- Email: [email protected]
Use of biotechnological approaches to explore the plant associated rhizobacteria
Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152
differences that have structural membrane component properties displaying up and down regulation and signalling. These findings open new avenues of research aimed at the target-discovery in oomycetes disease control by a new B. subtilis strain which is unique and that could be a suitable target for novel anti-oomycete agents. doi:10.1016/j.copbio.2011.05.084
OP14 Use of biotechnological approaches to explore the plant associated rhizobacteria Fauzia Yusuf Hafeez, Muhammad Nadeem Hassan Dept of Biosciences, COMSATS Institute of Information Technology, Islamabad, Pakistan E-mail address: [email protected] (F.Y. Hafeez) The bacteria inhabiting plant rhizosphere and endosphere play a vital role in improvement of plant health by providing nutrients and protecting them from various pathogens. Various cultural and biotechnological techniques were used to screen useful effective rhizobacteria. Among the thousands of screened strains, two potent strains, PCMC-1 and PCMC-2, solubilizing phosphate at 80–90 g mL−1 , maintaining a population level of 2 × 108 in humic acid up to 6–8 months and increasing plant growth in vivo, have been used to formulate commercial biofertilizers named Humiphos and Biophos. Exploration of sugarcane rhizosphere from various districts of Punjab, Pakistan, showed 2–16% prevalence of the antagonistic bacteria. Out of these, only 1% was found to be antibiotic producing bacteria. The antibiotic production was confirmed by various chromatographic and molecular techniques. The conserved genes involved in synthesis of antibiotics have facilitated the use of hybridization and PCR for the identification of these strains directly from the environment. In vivo studies have proved that the antibiotic producing strains are ideal candidates for the development of biopesticides. The results indicate that the use of advanced techniques is reliable to explore the microbes of plant rhizosphere and develop biopesticide/bioinoculants. doi:10.1016/j.copbio.2011.05.085
OP15 Improved bioluminescent detection of pesticides Ionescu Rodica Robert 2,3
Elena 1 , Jia
Kun 1 , Eltzov
Evgeni 2,3 , Marks
1 Laboratoire de Nanotechnologie et d’Instrumentation Optique, Institute Charles Delaunay, Université de Technologie de Troyes, Troyes, France 2 Department of Environmental Engineering, Faculty of Engineering Science, Ben-Gurion University of the Negev, Beer-Sheva, Israel 3 National Institute for Biotechnology in the Negev Ben-Gurion University of the Negev, Beer-Sheva, Israel
E-mail address: [email protected] (I.R. Elena) Toxicity of three common poorly water soluble agricultural pollutants atrazine, nonyl-phenol and carbofuran were investigated using two engineered recombinant bacterial E. coli strains — TV1061 and DPD2794 — harbouring light-emitting reporter genes such lux. Cultivation of bacteria cells was performed in Luria-Bertani (LB) medium supplemented with kanamycin antibiotic at 37◦ C for 9 hours (overnight), while the re-growth steps to bacteria early exponential phases (0.02, 0.03,
S37
0.08, 0.12, 0.20) at OD600 were acquired in LB media without antibiotic. High toxicant concentrations (10–1 g/ml) and low toxicant concentrations (100 pg/ml–10 fg/ml) were mixed with bacterial strains using 3 mixing procedures (manual, vortex, and centrifuge) while the resulted bioluminescence signals were investigated at different temperatures (4◦ C, 17◦ C, 25◦ C, 30◦ C). Interesting experimental bioluminescence data such as signal intensity, shape and appearance were acquired attesting very particular bacterial metabolic behaviours in the presence of the selected pesticides. For all mixing procedures, the overnight ‘toxic stress’, bacteria gave a maximum bioluminescence signals in a short time (20–30 min) while for fresh ‘toxic stress’ maximum bioluminescence peak appeared after 180–300 min. Moreover, we found that the best measurements for “high” and “low” toxicant detection are made when using very early exponential growth of bacteria such as 0.02 followed by incubation at 4◦ C for a period of 9 hours and luminescence reading spectra at 25◦ C using centrifuge and vortex approaches. doi:10.1016/j.copbio.2011.05.086
OP16 Phytoremediation of cadmium polluted soils from phosphorus fertilizers Safoora Asadi Kapourchal 1 , Somayyeh Eisazadeh Lazarjan 2 , Mehdi Homaee 3 1
Member of Young Researchers Club, Science and Research Branch, Islamic Azad University, Tehran, Iran 2 Soil Science Department, Science and Research Branch, Islamic Azad University, Tehran, Iran 3 Soil Science Department, Tarbiat Modares University, Tehran, Iran E-mail address: [email protected] (S.A. Kapourchal) One technology to remediate contaminated soils is the so-called phytoextraction. Phytoextraction technology can be considered as a sustainable, environment-friendly and inexpensive technique. It has minimum destructive impact on the ecosystem. The objective of this study was to investigate the extraction of Cadmium from soils contaminated with cadmium resulting from phosphorous fertilizers with spinach and chive. A randomized block experiment design was performed. The soil was contaminated with CdNO3 and treatments consisted of 0.663 (standard), 15, 30, 60 and120 mg/kg cadmium. After development, plants were harvested and divided into shoots and roots. The cadmium content of each plants part as well as the soil-cadmium was measured. The results indicated a non-linear positive relation between the cadmium concentrations in soil and that accumulated in plant parts. By increasing the cadmium concentration in soil, its accumulation in plant tissues was increased and its accumulation in roots was more than shoots. Most of the extracted cadmium for spinach and chive was accumulated in the roots, 75.86 and 482.48 mg/kg respectively. Finally, chive’s capability was more than spinach for phytoextraction of cadmium. doi:10.1016/j.copbio.2011.05.087
differences that have structural membrane component properties displaying up and down regulation and signalling. These findings open new avenues of research aimed at the target-discovery in oomycetes disease control by a new B. subtilis strain which is unique and that could be a suitable target for novel anti-oomycete agents. doi:10.1016/j.copbio.2011.05.084
OP14 Use of biotechnological approaches to explore the plant associated rhizobacteria Fauzia Yusuf Hafeez, Muhammad Nadeem Hassan Dept of Biosciences, COMSATS Institute of Information Technology, Islamabad, Pakistan E-mail address: [email protected] (F.Y. Hafeez) The bacteria inhabiting plant rhizosphere and endosphere play a vital role in improvement of plant health by providing nutrients and protecting them from various pathogens. Various cultural and biotechnological techniques were used to screen useful effective rhizobacteria. Among the thousands of screened strains, two potent strains, PCMC-1 and PCMC-2, solubilizing phosphate at 80–90 g mL−1 , maintaining a population level of 2 × 108 in humic acid up to 6–8 months and increasing plant growth in vivo, have been used to formulate commercial biofertilizers named Humiphos and Biophos. Exploration of sugarcane rhizosphere from various districts of Punjab, Pakistan, showed 2–16% prevalence of the antagonistic bacteria. Out of these, only 1% was found to be antibiotic producing bacteria. The antibiotic production was confirmed by various chromatographic and molecular techniques. The conserved genes involved in synthesis of antibiotics have facilitated the use of hybridization and PCR for the identification of these strains directly from the environment. In vivo studies have proved that the antibiotic producing strains are ideal candidates for the development of biopesticides. The results indicate that the use of advanced techniques is reliable to explore the microbes of plant rhizosphere and develop biopesticide/bioinoculants. doi:10.1016/j.copbio.2011.05.085
OP15 Improved bioluminescent detection of pesticides Ionescu Rodica Robert 2,3
Elena 1 , Jia
Kun 1 , Eltzov
Evgeni 2,3 , Marks
1 Laboratoire de Nanotechnologie et d’Instrumentation Optique, Institute Charles Delaunay, Université de Technologie de Troyes, Troyes, France 2 Department of Environmental Engineering, Faculty of Engineering Science, Ben-Gurion University of the Negev, Beer-Sheva, Israel 3 National Institute for Biotechnology in the Negev Ben-Gurion University of the Negev, Beer-Sheva, Israel
E-mail address: [email protected] (I.R. Elena) Toxicity of three common poorly water soluble agricultural pollutants atrazine, nonyl-phenol and carbofuran were investigated using two engineered recombinant bacterial E. coli strains — TV1061 and DPD2794 — harbouring light-emitting reporter genes such lux. Cultivation of bacteria cells was performed in Luria-Bertani (LB) medium supplemented with kanamycin antibiotic at 37◦ C for 9 hours (overnight), while the re-growth steps to bacteria early exponential phases (0.02, 0.03,
S37
0.08, 0.12, 0.20) at OD600 were acquired in LB media without antibiotic. High toxicant concentrations (10–1 g/ml) and low toxicant concentrations (100 pg/ml–10 fg/ml) were mixed with bacterial strains using 3 mixing procedures (manual, vortex, and centrifuge) while the resulted bioluminescence signals were investigated at different temperatures (4◦ C, 17◦ C, 25◦ C, 30◦ C). Interesting experimental bioluminescence data such as signal intensity, shape and appearance were acquired attesting very particular bacterial metabolic behaviours in the presence of the selected pesticides. For all mixing procedures, the overnight ‘toxic stress’, bacteria gave a maximum bioluminescence signals in a short time (20–30 min) while for fresh ‘toxic stress’ maximum bioluminescence peak appeared after 180–300 min. Moreover, we found that the best measurements for “high” and “low” toxicant detection are made when using very early exponential growth of bacteria such as 0.02 followed by incubation at 4◦ C for a period of 9 hours and luminescence reading spectra at 25◦ C using centrifuge and vortex approaches. doi:10.1016/j.copbio.2011.05.086
OP16 Phytoremediation of cadmium polluted soils from phosphorus fertilizers Safoora Asadi Kapourchal 1 , Somayyeh Eisazadeh Lazarjan 2 , Mehdi Homaee 3 1
Member of Young Researchers Club, Science and Research Branch, Islamic Azad University, Tehran, Iran 2 Soil Science Department, Science and Research Branch, Islamic Azad University, Tehran, Iran 3 Soil Science Department, Tarbiat Modares University, Tehran, Iran E-mail address: [email protected] (S.A. Kapourchal) One technology to remediate contaminated soils is the so-called phytoextraction. Phytoextraction technology can be considered as a sustainable, environment-friendly and inexpensive technique. It has minimum destructive impact on the ecosystem. The objective of this study was to investigate the extraction of Cadmium from soils contaminated with cadmium resulting from phosphorous fertilizers with spinach and chive. A randomized block experiment design was performed. The soil was contaminated with CdNO3 and treatments consisted of 0.663 (standard), 15, 30, 60 and120 mg/kg cadmium. After development, plants were harvested and divided into shoots and roots. The cadmium content of each plants part as well as the soil-cadmium was measured. The results indicated a non-linear positive relation between the cadmium concentrations in soil and that accumulated in plant parts. By increasing the cadmium concentration in soil, its accumulation in plant tissues was increased and its accumulation in roots was more than shoots. Most of the extracted cadmium for spinach and chive was accumulated in the roots, 75.86 and 482.48 mg/kg respectively. Finally, chive’s capability was more than spinach for phytoextraction of cadmium. doi:10.1016/j.copbio.2011.05.087