The genome sequence of T. parva shows significant differences from other apicomplexan genomes sequenced to date. It provides improvements in understanding T. parva's metabolic capabilities and a foundation for studying host cell transformation, which is important for a pathogen of agricultural significance in Africa. Mining the sequence data has already proved useful in searching for candidate vaccine antigens.
Effect of plant growth promoting rhizobacterial (PGPR) inoculation on growth ...IJEAB
Plant Growth promoting rhizobacteria are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots. They benefit plants through Production of plant hormones, such as auxins, asymbiotic N2 fixation, solubilization of mineral phosphates, antagonism against phytopathogenic microorganisms by production of antibiotics, siderophroes, Chitinase and other nutrients ability to effectively colonize roots are responsible for plant growth promotion. An experiment was conducted in the field of National Institute of Agronomic Research of Meknes. Morocco. The experiment was a completely randomized design with six replicates. There were four treatments viz. T1: (control; N0 -PGPR), T2: (N0 +2027-2), T3: (N0 +2066-7) and T4: (N0+2025-1). The results indicated that a remarkable increase in root growth, namely length, the diameter of the rod and the total chlorophyll. A total of three different bacteria colonies were isolated and proceed with in vitro screening for plant growth promoting activities; phosphate solubilization, nitrogen fixation, indole acetic acid (IAA), ammonia production and antimicrobial enzymes (cellulose, chitinase and protease) activity. Among the three bacterial strains, all bacterial strains are able to produce ammonia, IAA production and nitrogen fixation activity, one strain phosphate solubilizing activity, two strain are able to produce cellulase syntheses, Protease activity and Chitinase activity.
Effect of sucrose on inducing in vitro microtuberization in potato without us...Innspub Net
The present in vitro experiment was conducted to investigate the effect of the various concentrations of sucrose on potato plantlets growth and microtuberization. It was observed that increasing sucrose level in the media influenced the plant growth negatively. 3% sucrose concentration in the medium showed comparatively early root/shoot emergence and highest mean root and shoot length (6.16 cm and 8.28 cm, respectively) with greater number of nodes (7.90). However, regarding microtubers (Mt) formation, treatment with 8% sucrose concentration has higher microtubers number with larger size (mean diameter 6.84mm). The mean weight of Mt
was also highest (97.0mg) at 8 % sucrose concentration followed by T1 (70.00mg). It has been concluded on the
basis of results that MS medium supplemented with 8% sucrose level and without any growth hormone is the best
for in vitro microtuber formation in potato.
Investigate different measurements of cellulase screening and the effect of different substrates under solid-state fermentation on conidia production or enzyme activity by Trichoderma
Effect of plant growth promoting rhizobacterial (PGPR) inoculation on growth ...IJEAB
Plant Growth promoting rhizobacteria are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots. They benefit plants through Production of plant hormones, such as auxins, asymbiotic N2 fixation, solubilization of mineral phosphates, antagonism against phytopathogenic microorganisms by production of antibiotics, siderophroes, Chitinase and other nutrients ability to effectively colonize roots are responsible for plant growth promotion. An experiment was conducted in the field of National Institute of Agronomic Research of Meknes. Morocco. The experiment was a completely randomized design with six replicates. There were four treatments viz. T1: (control; N0 -PGPR), T2: (N0 +2027-2), T3: (N0 +2066-7) and T4: (N0+2025-1). The results indicated that a remarkable increase in root growth, namely length, the diameter of the rod and the total chlorophyll. A total of three different bacteria colonies were isolated and proceed with in vitro screening for plant growth promoting activities; phosphate solubilization, nitrogen fixation, indole acetic acid (IAA), ammonia production and antimicrobial enzymes (cellulose, chitinase and protease) activity. Among the three bacterial strains, all bacterial strains are able to produce ammonia, IAA production and nitrogen fixation activity, one strain phosphate solubilizing activity, two strain are able to produce cellulase syntheses, Protease activity and Chitinase activity.
Effect of sucrose on inducing in vitro microtuberization in potato without us...Innspub Net
The present in vitro experiment was conducted to investigate the effect of the various concentrations of sucrose on potato plantlets growth and microtuberization. It was observed that increasing sucrose level in the media influenced the plant growth negatively. 3% sucrose concentration in the medium showed comparatively early root/shoot emergence and highest mean root and shoot length (6.16 cm and 8.28 cm, respectively) with greater number of nodes (7.90). However, regarding microtubers (Mt) formation, treatment with 8% sucrose concentration has higher microtubers number with larger size (mean diameter 6.84mm). The mean weight of Mt
was also highest (97.0mg) at 8 % sucrose concentration followed by T1 (70.00mg). It has been concluded on the
basis of results that MS medium supplemented with 8% sucrose level and without any growth hormone is the best
for in vitro microtuber formation in potato.
Investigate different measurements of cellulase screening and the effect of different substrates under solid-state fermentation on conidia production or enzyme activity by Trichoderma
The objectives of this study are: (i): To investigate and recognize the internal and abnormalities impacts induced by phytoplasma infection in the tomato host according to recent studies have shown that the association between plants and phytoplasmas can result in anatomical alteration in phloem tissues of infected plants, and great differences between healthy and diseased samples using microscopic examination of longitudinal, cross or ultra-thin sections of leaf blade, leaf petiole and stem. (ii): To determine the efficiency of different techniques toward production of phytoplasma-free tomato plantlets and mitigation of phytoplasma disease.
Ecklonia Maxima Extract Effect in Tissue Regeneration of Symbionts at in Vivo...IJEAB
The research was conducted to determine the Ecklonia maxima extract effect in the symbiont accretion at the in vivo heteroplasmic grafting of some tomatoes. E. maxima or sea bamboo is a seaweed used for obtaining of organic extracts used as stimulators in horticulture because consists the natural plant hormones such as auxins and cytokinins which have optimal role in cell division, important activity for tissue regeneration. The experimental variants were grafted plants, combinations between different symbionts, cultivar fragments from Lycopersicon esculentum specie. The symbionts were two scions, F1 hybrids, 'Siriana' (Romanian cultivar), 'Abellus' (Dutch cultivar) and two rootstocks, 'Buzău' variety (Romanian cultivar), 'Emperador' F1 hybrid (Dutch cultivar). The algae extract used had auxins (11 mg/L) and cytokinins (0.3 mg/L). Two treatments were applied before grafting on scion and rootstock, 1 mL/500 mL water and a treatment at grafting on soil, 5 mL/L water. Control variant was without hormone extract. Determinations, observations and interpretations of the algae effect were made on symbionts. The best results on tissue regeneration were obtained in plants treated with sea bamboo extract compared to untreated control plants. The E. maxima extract influenced the tissue regeneration.
Slide show..prof.s.p.singh.invited talk. udapur international conference.02 j...Saurashtra University
In Invited Talk by Prof. Satya P. Singh on:
Attributes of Plant Growth Promoting Rhizobacteria in biocontrol and combating salinity stress
An International Conference on Recent Trends in Plant Sciences held at M L Sukhadia University, Udaipur, Rajasthan, India
Effect of Plastic Mulch, Floating Row Cover & Microtunnels on Insect Populations & Yield of Muskmelon; Gardening Guidebook for Mexico www.scribd.com/doc/239851313 ~ Universidad de Colima, For more information, Please see Organic Edible Schoolyards & Gardening with Children www.scribd.com/doc/239851214, Double Food Production from your School Garden with Organic Tech www.scribd.com/doc/239851079, Free School Gardening Art Posters www.scribd.com/doc/239851159, Increase Food Production with Companion Planting in your School Garden www.scribd.com/doc/239851159, Healthy Foods Dramatically Improves Student Academic Success www.scribd.com/doc/239851348, City Chickens for your Organic School Garden www.scribd.com/doc/239850440, Huerto Ecológico, Tecnologías Sostenibles, Agricultura Organica www.scribd.com/doc/239850233, Simple Square Foot Gardening for Schools - Teacher Guide www.scribd.com/doc/23985111 ~
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
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.
Somatic Embryogenesis in Pearl Millet (Pennisetum Glaucum (L) R.Br.) Using Li...IOSR Journals
The morphology and histology of somatic embryogenesis were examined using embryogenic calli derived from immature inflorescences of pearl millet using light and electron microscopic techniques. Young immature inflorescences, 30-35 days old (2.5 cm long) were cultured on LS medium supplemented with 2.0 mgL-1 of 2, 4-D and 0.5 mgL-1 of kinetin. The primary callus initiated was non-embryogenic, loose, white but differentiated rapidly into pale yellow, nodular and friable embryogenic callus within 2 weeks. Histological studies revealed the formation of somatic embryos and their maturation. Initial stages of somatic embryo induction showed calli with internal segmenting divisions, which gave rise to discrete groups of cells. Continued divisions and organization in some discrete groups formed globular structures, presumably proembryoids, which showed a distinct epidermis. Further these structures developed to form somatic embryos with a well-organized bipolar structure showing embryonic axis, scutellum, coleoptile and coleorhiza. Multiple shoots (5-6) were produced from each embryo when transferred to MS regeneration medium supplemented with hormonal combinations, viz., BAP + IBA, 2.0 + 0.5 mg L-1 which further developed into complete plantlets
The objectives of this study are: (i): To investigate and recognize the internal and abnormalities impacts induced by phytoplasma infection in the tomato host according to recent studies have shown that the association between plants and phytoplasmas can result in anatomical alteration in phloem tissues of infected plants, and great differences between healthy and diseased samples using microscopic examination of longitudinal, cross or ultra-thin sections of leaf blade, leaf petiole and stem. (ii): To determine the efficiency of different techniques toward production of phytoplasma-free tomato plantlets and mitigation of phytoplasma disease.
Ecklonia Maxima Extract Effect in Tissue Regeneration of Symbionts at in Vivo...IJEAB
The research was conducted to determine the Ecklonia maxima extract effect in the symbiont accretion at the in vivo heteroplasmic grafting of some tomatoes. E. maxima or sea bamboo is a seaweed used for obtaining of organic extracts used as stimulators in horticulture because consists the natural plant hormones such as auxins and cytokinins which have optimal role in cell division, important activity for tissue regeneration. The experimental variants were grafted plants, combinations between different symbionts, cultivar fragments from Lycopersicon esculentum specie. The symbionts were two scions, F1 hybrids, 'Siriana' (Romanian cultivar), 'Abellus' (Dutch cultivar) and two rootstocks, 'Buzău' variety (Romanian cultivar), 'Emperador' F1 hybrid (Dutch cultivar). The algae extract used had auxins (11 mg/L) and cytokinins (0.3 mg/L). Two treatments were applied before grafting on scion and rootstock, 1 mL/500 mL water and a treatment at grafting on soil, 5 mL/L water. Control variant was without hormone extract. Determinations, observations and interpretations of the algae effect were made on symbionts. The best results on tissue regeneration were obtained in plants treated with sea bamboo extract compared to untreated control plants. The E. maxima extract influenced the tissue regeneration.
Slide show..prof.s.p.singh.invited talk. udapur international conference.02 j...Saurashtra University
In Invited Talk by Prof. Satya P. Singh on:
Attributes of Plant Growth Promoting Rhizobacteria in biocontrol and combating salinity stress
An International Conference on Recent Trends in Plant Sciences held at M L Sukhadia University, Udaipur, Rajasthan, India
Effect of Plastic Mulch, Floating Row Cover & Microtunnels on Insect Populations & Yield of Muskmelon; Gardening Guidebook for Mexico www.scribd.com/doc/239851313 ~ Universidad de Colima, For more information, Please see Organic Edible Schoolyards & Gardening with Children www.scribd.com/doc/239851214, Double Food Production from your School Garden with Organic Tech www.scribd.com/doc/239851079, Free School Gardening Art Posters www.scribd.com/doc/239851159, Increase Food Production with Companion Planting in your School Garden www.scribd.com/doc/239851159, Healthy Foods Dramatically Improves Student Academic Success www.scribd.com/doc/239851348, City Chickens for your Organic School Garden www.scribd.com/doc/239850440, Huerto Ecológico, Tecnologías Sostenibles, Agricultura Organica www.scribd.com/doc/239850233, Simple Square Foot Gardening for Schools - Teacher Guide www.scribd.com/doc/23985111 ~
— The microbiological content of Lettuce (a vegetable), commonly vended in the Benin metropolis of Edo state were evaluated. Five vending locations were chosen for the study. Whole and soft rot samples were purchased and analysed for microbiological composition. Results showed high counts in soft rot samples in lettuce. Nutrient agar plated lettuce samples had bacterial counts in the range of 2.0x 103 to 4.7x10 7. Pseudomonas species was the dominant species found in lettuce samples. Bacillus species was isolated from one location in the lettuce samples. Mac Conkey agar plated lettuce plated had bacterial counts in the range of 2.3 x 10 3 to 5.7x 10 7. Enterobacter species, E. coli, and Klebsiella species were the dominant species isolated. Though, Proteus species was isolated from lettuce samples obtained from location five only. The study observes that consuming soft rot samples could pose a risk of introducing pathogens to the consumer due to their high microbial counts and could be detrimental to the health of the consumer.
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.
Somatic Embryogenesis in Pearl Millet (Pennisetum Glaucum (L) R.Br.) Using Li...IOSR Journals
The morphology and histology of somatic embryogenesis were examined using embryogenic calli derived from immature inflorescences of pearl millet using light and electron microscopic techniques. Young immature inflorescences, 30-35 days old (2.5 cm long) were cultured on LS medium supplemented with 2.0 mgL-1 of 2, 4-D and 0.5 mgL-1 of kinetin. The primary callus initiated was non-embryogenic, loose, white but differentiated rapidly into pale yellow, nodular and friable embryogenic callus within 2 weeks. Histological studies revealed the formation of somatic embryos and their maturation. Initial stages of somatic embryo induction showed calli with internal segmenting divisions, which gave rise to discrete groups of cells. Continued divisions and organization in some discrete groups formed globular structures, presumably proembryoids, which showed a distinct epidermis. Further these structures developed to form somatic embryos with a well-organized bipolar structure showing embryonic axis, scutellum, coleoptile and coleorhiza. Multiple shoots (5-6) were produced from each embryo when transferred to MS regeneration medium supplemented with hormonal combinations, viz., BAP + IBA, 2.0 + 0.5 mg L-1 which further developed into complete plantlets
The inhibitory activity of L. crispatus against uropathogenes in vitroIJMCERJournal
ABSTRACT: Bacterial interference refers to the use of bacteria of virulence to compete with and protect against
colonization and infection by disease causing organisms. In this study, L. crispatus strain was used to detect its
antibacterial activity towered five species (one Gram-positive and four Gram-negative ) of the most common bacteria
causing urinary tract infection. L. crispatus completely inhibited growth of Staphylococcus aureus , while it had no
inhibitory effect on three of the other species.
Key Words: inhibitory activity, L. crispatus, uropathogens
Respiration of e. coli in the mouse intestineAndrew Fabich
Mammals are aerobes that harbor an intestinal ecosystem dominated by large numbers of anaerobic microorganisms. However, the role of oxygen in the intestinal ecosystem is largely unexplored. We used systematic mutational analysis to determine the role of respiratory metabolism in the streptomycin-treated mouse model of intestinal colonization. Here we provide evidence that aerobic respiration is required for commensal and pathogenic Escherichia coli to colonize mice. Our results showed that mutants lacking ATP synthase, which is required for all respiratory energy-conserving metabolism, were eliminated by competition with respiratory-competent wild-type strains. Mutants lacking the high-affinity cytochrome bd oxidase, which is used when oxygen tensions are low, also failed to colonize. However, the low-affinity cytochrome bo(3) oxidase, which is used when oxygen tension is high, was found not to be necessary for colonization. Mutants lacking either nitrate reductase or fumarate reductase also had major colonization defects. The results showed that the entire E. coli population was dependent on both microaerobic and anaerobic respiration, consistent with the hypothesis that the E. coli niche is alternately microaerobic and anaerobic, rather than static. The results indicate that success of the facultative anaerobes in the intestine depends on their respiratory flexibility. Despite competition for relatively scarce carbon sources, the energy efficiency provided by respiration may contribute to the widespread distribution (i.e., success) of E. coli strains as commensal inhabitants of the mammalian intestine.
Proteomic analysis of the interaction between the plant growth promoting fhiz...kys9723331
Plant growth-promoting rhizobacteria (PGPR) facilitate the plant growth and enhance their
induced systemic resistance (ISR) against a variety of environmental stresses. In this study,
we carried out integrative analyses on the proteome, transcriptome, and metabolome to investigate
Arabidopsis root and shoot responses to the well-known PGPR strain Paenibacillus
polymyxa (P. polymyxa) E681. Shoot fresh and root dry weights were increased, whereas root
length was decreased by treatment with P. polymyxa E681. 2DE approach in conjunction
with MALDI-TOF/TOF analysis revealed a total of 41 (17 spots in root, 24 spots in shoot)
that were differentially expressed in response to P. polymyxa E681. Biological process- and
molecular function-based bioinformatics analysis resulted in their classification into seven different
protein groups. Of these, 36 proteins including amino acid metabolism, antioxidant,
defense and stress response, photosynthesis, and plant hormone-related proteins were upregulated,
whereas five proteins including three carbohydrate metabolism- and one amino
acid metabolism-related, and one unknown protein were down-regulated, respectively. A good
correlation was observed between protein and transcript abundances for the 12 differentially
expressed proteins during interactions as determined by qPCR analysis. Metabolite analysis
using LC-MS/MS revealed highly increased levels of tryptophan, indole-3-acetonitrile (IAN),
indole-3-acetic acid (IAA), and camalexin in the treated plants. Arabidopsis plant inoculated
P. polymyxa E681 also showed resistance to Botrytis cinerea infection. Taken together these
results suggest that P. polymyxa E681 may promote plant growth by induced metabolism and
activation of defense-related proteins against fungal pathogen.
"Keeping up with the plant destroyers." My talk at The Royal Society, 7 March...Sophien Kamoun
Tackling emerging threats to animal health, food security and ecosystem resilience, The Royal Society, Monday 7 – Tuesday 8 March 2016. https://royalsociety.org/events/2016/03/emerging-fungal-threats/
My talk at BASF Science Symposium: sustainable food chain - from field to table, Jun 23-24, 2015, Chicago.
Notes and acknowledgements at http://kamounlab.tumblr.com/post/122151022390/plant-pathology-in-the-post-genomics-era
Role of Phytohormones in Tissue CultureApoorva Ashu
Description about phytohormones and their role in tissue culture, including descriptions about molecular basis of phytohormones with special focus on auxin and cytokinin and their role in calli development, organogenesis and somatic embryogenesis.
nduced Mutation by Colchicine Treatment of Somatic Embryos in ‘Namwa’ Banana ...drboon
Hexaploids of the ‘Namwa’ banana (Musa sp ABB) were obtained by in vitro colchicine treatment of somatic embryos. Somatic embryos were induced on a medium containing MS medium supplemented with 8 mg/l picloram. Somatic embryos were treated with four different concentrations of colchicine (0, 0.3, 0.5, 1.0, %) in liquid MS medium supplemented with 0.22 mg/l zeatin, and shaken (60 rpm) at 25 0C in darkness for 48, 72 or 96 hours. Higher colchicine concentrations exhibited higher mortality rates ranging from 8–20%, 48–62% and 80–90% mortality on concentrations 0.3, 0.5, and 1.0 % colchicine respectively. Mortality rate generally increased with increased treatment time. Hexaploids were obtained at a frequency of 2 % with treatments 0.05 % colchicine for 96 hours, and 1 % colchicine for 48 hours as determined by flow cytometry.
I have discussed Applications of Plant Tissue Culture under the following subheadings,
1. Micro Propagation
2. Clonal Propagation
3. Production of Genetically Variable Plants
4. Production of Virus Free Plants
5. Plant Breeding
6. Production of Useful Biochemicals
7. Preservation of Plant Genetic Resources
8. Importance of Tissue Culture in Biotechnology
Industrial Training at Shahjalal Fertilizer Company Limited (SFCL)MdTanvirMahtab2
This presentation is about the working procedure of Shahjalal Fertilizer Company Limited (SFCL). A Govt. owned Company of Bangladesh Chemical Industries Corporation under Ministry of Industries.
Final project report on grocery store management system..pdfKamal Acharya
In today’s fast-changing business environment, it’s extremely important to be able to respond to client needs in the most effective and timely manner. If your customers wish to see your business online and have instant access to your products or services.
Online Grocery Store is an e-commerce website, which retails various grocery products. This project allows viewing various products available enables registered users to purchase desired products instantly using Paytm, UPI payment processor (Instant Pay) and also can place order by using Cash on Delivery (Pay Later) option. This project provides an easy access to Administrators and Managers to view orders placed using Pay Later and Instant Pay options.
In order to develop an e-commerce website, a number of Technologies must be studied and understood. These include multi-tiered architecture, server and client-side scripting techniques, implementation technologies, programming language (such as PHP, HTML, CSS, JavaScript) and MySQL relational databases. This is a project with the objective to develop a basic website where a consumer is provided with a shopping cart website and also to know about the technologies used to develop such a website.
This document will discuss each of the underlying technologies to create and implement an e- commerce website.
Hierarchical Digital Twin of a Naval Power SystemKerry Sado
A hierarchical digital twin of a Naval DC power system has been developed and experimentally verified. Similar to other state-of-the-art digital twins, this technology creates a digital replica of the physical system executed in real-time or faster, which can modify hardware controls. However, its advantage stems from distributing computational efforts by utilizing a hierarchical structure composed of lower-level digital twin blocks and a higher-level system digital twin. Each digital twin block is associated with a physical subsystem of the hardware and communicates with a singular system digital twin, which creates a system-level response. By extracting information from each level of the hierarchy, power system controls of the hardware were reconfigured autonomously. This hierarchical digital twin development offers several advantages over other digital twins, particularly in the field of naval power systems. The hierarchical structure allows for greater computational efficiency and scalability while the ability to autonomously reconfigure hardware controls offers increased flexibility and responsiveness. The hierarchical decomposition and models utilized were well aligned with the physical twin, as indicated by the maximum deviations between the developed digital twin hierarchy and the hardware.
Saudi Arabia stands as a titan in the global energy landscape, renowned for its abundant oil and gas resources. It's the largest exporter of petroleum and holds some of the world's most significant reserves. Let's delve into the top 10 oil and gas projects shaping Saudi Arabia's energy future in 2024.
NO1 Uk best vashikaran specialist in delhi vashikaran baba near me online vas...Amil Baba Dawood bangali
Contact with Dawood Bhai Just call on +92322-6382012 and we'll help you. We'll solve all your problems within 12 to 24 hours and with 101% guarantee and with astrology systematic. If you want to take any personal or professional advice then also you can call us on +92322-6382012 , ONLINE LOVE PROBLEM & Other all types of Daily Life Problem's.Then CALL or WHATSAPP us on +92322-6382012 and Get all these problems solutions here by Amil Baba DAWOOD BANGALI
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Student information management system project report ii.pdfKamal Acharya
Our project explains about the student management. This project mainly explains the various actions related to student details. This project shows some ease in adding, editing and deleting the student details. It also provides a less time consuming process for viewing, adding, editing and deleting the marks of the students.
Welcome to WIPAC Monthly the magazine brought to you by the LinkedIn Group Water Industry Process Automation & Control.
In this month's edition, along with this month's industry news to celebrate the 13 years since the group was created we have articles including
A case study of the used of Advanced Process Control at the Wastewater Treatment works at Lleida in Spain
A look back on an article on smart wastewater networks in order to see how the industry has measured up in the interim around the adoption of Digital Transformation in the Water Industry.
1. The genome sequence of T. parva shows
remarkable differences from the other apicom-
plexan genomes sequenced to date. It provides
significant improvements in our understanding
of the metabolic capabilities of T. parva and a
foundation for studying parasite-induced host
cell transformation and constitutes a critical
knowledge base for a pathogen of significance
to agriculture in Africa. Mining of sequence
data has already proved useful in the search for
candidate vaccine antigens (3).
References and Notes
1. R. A. I. Norval, B. D. Perry, A. S. Young, The Epi-
demiology of Theileriosis in Africa (Academic Press,
London, 1992), p. 481.
2. A. A. Escalante, F. J. Ayala, Proc. Natl. Acad. Sci.
U.S.A. 92, 5793 (1995).
3. S. P. Graham et al., in preparation.
4. A. Pain et al., Science 309, 131 (2005).
5. Materials and methods are available as supporting
material on Science Online.
6. A. Kairo, A. H. Fairlamb, E. Gobright, V. Nene, EMBO
J. 13, 898 (1994).
7. M. J. Gardner et al., Nature 419, 498 (2002).
8. J. H. Gunderson et al., Science 238, 933 (1987).
9. B. Vogelstein, K. W. Kinzler, Nat. Med. 10, 789 (2004).
10. M. Goedert, Semin. Cell Dev. Biol. 15, 45 (2004).
11. I. M. Cheeseman, A. Desai, Curr. Biol. 14, R70 (2004).
12. R. F. Anders, Parasite Immunol. 8, 529 (1986).
13. R. Bishop et al., Mol. Biochem. Parasitol. 110, 359
(2000).
14. B. Sohanpal, D. Wasawo, R. Bishop, Gene 255, 401
(2000).
15. H. A. Baylis, S. K. Sohal, M. Carrington, R. P. Bishop, B. A.
Allsopp, Mol. Biochem. Parasitol. 49, 133 (1991).
16. J. D. Barry, M. L. Ginger, P. Burton, R. McCulloch, Int.
J. Parasitol. 33, 29 (2003).
17. R. Bishop et al., in preparation.
18. Z. Su et al., Cell 82, 89 (1995).
19. R. Bishop, A. Musoke, S. Morzaria, B. Sohanpal, E.
Gobright, Mol. Cell. Biol. 17, 1666 (1997).
20. M. S. Abrahamsen et al., Science 304, 441 (2004);
published online 25 March 2004 (10.1126/science.
1094786).
21. B. J. Foth et al., Mol. Microbiol. 55, 39 (2005).
22. S. A. Ralph, Mol. Microbiol. 55, 1 (2005).
23. A. A. McColm, N. McHardy, Ann. Trop. Med. Para-
sitol. 78, 345 (1984).
24. R. F. Waller, G. I. McFadden, Curr. Issues Mol. Biol. 7,
57 (2005).
25. R. F. Waller et al., Proc. Natl. Acad. Sci. U.S.A. 95,
12352 (1998).
26. F. Seeber, Int. J. Parasitol. 32, 1207 (2002).
27. X. M. Xu, S. G. Moller, Proc. Natl. Acad. Sci. U.S.A.
101, 9143 (2004).
28. S. Leon, B. Touraine, C. Ribot, J. F. Briat, S. Lobreaux,
Biochem. J. 371, 823 (2003).
29. J. M. Carlton et al., Nature 419, 512 (2002).
30. R. J. Wilson et al., J. Mol. Biol. 261, 155 (1996).
31. Single-letter abbreviations for the amino acid residues
are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G,
Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q,
Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
32. We thank T. Irvin, O. Ole-MoiYoi, T. Musoke, C.
Sugimoto, H. Leitch, R. von Kaufmann, S. MacMillan,
R. Koenig, M. Brown, R. Ndegwa, L. Thairo, B. Anyona,
T. Akinyemi, the TIGR conferences staff, the Secre-
tariat of the Consultative Group for International
Agricultural Research, and the research staff of the
International Livestock Research Institute (ILRI).
Supported by the TIGR Board of Trustees, ILRI, J. C.
Venter, the Rockefeller Foundation, the U.S. Agency
for International Development, and the UK Depart-
ment for International Development.
Supporting Online Material
www.sciencemag.org/cgi/content/full/309/5731/134/
DC1
Materials and Methods
Figs. S1 to S3
Tables S1 to S3
31 January 2005; accepted 5 May 2005
10.1126/science.1110439
Long-Term Monitoring of Bacteria
Undergoing ProgrammedPopulation
Control in a Microchemostat
Frederick K. Balagadde´,1
*. Lingchong You,2
.- Carl L. Hansen,1
`
Frances H. Arnold,2
Stephen R. Quake1
*¬
Using an active approach to preventing biofilm formation, we implemented a
microfluidic bioreactor that enables long-term culture and monitoring of extremely
small populations of bacteria with single-cell resolution. We used this device to
observe the dynamics of Escherichia coli carrying a synthetic ‘‘population control’’
circuit that regulates cell density through a feedback mechanism based on quorum
sensing. The microfluidic bioreactor enabled long-term monitoring of unnatural
behavior programmed by the synthetic circuit, which included sustained oscilla-
tions in cell density and associated morphological changes, over hundreds of hours.
By continually substituting a fraction of a
bacterial culture with sterile nutrients, the
chemostat (1, 2) presents a near-constant en-
vironment that is ideal for controlled studies of
microbes and microbial communities (3–6).
The considerable challenges of maintaining
and operating continuous bioreactors, includ-
ing the requirement for large quantities of
growth media and reagents, have pushed the
move toward miniaturization and chip-based
control (7–10), although efforts have been
limited to batch-format operation. Microbial
biofilms, which exist in virtually all nutrient-
sufficient ecosystems (11), interfere with
continuous bioreactor operation (12). Pheno-
typically distinct from their planktonic coun-
terparts (11), biofilm cells shed their progeny
into the bulk culture and create mixed cultures.
At high dilution rates, the biofilm, which is not
subject to wash-out, supplies most of the bulk-
culture cells (13). The increase in surface area-to-
volume ratio as the working volume is decreased
aggravates these wall-growth effects (13).
We created a chip-based bioreactor that
uses microfluidic plumbing networks to ac-
tively prevent biofilm formation. This device
allows semicontinuous, planktonic growth in
six independent 16-nanoliter reactors with no
observable wall growth (Fig. 1A). The cultures
can be monitored in situ by optical microscopy
to provide automated, real-time, noninvasive
measurement of cell density and morphology
with single-cell resolution.
Each reactor, or Bmicrochemostat,[ con-
sists of a growth chamber, which is a fluidic
loop 10 mm high, 140 mm wide, and 11.5 mm
in circumference, with an integrated peristaltic
pump and a series of micromechanical valves
to add medium, remove waste, and recover
cells (Fig. 1B). The growth loop is itself
composed of 16 individually addressable seg-
ments. The microchemostat is operated in one
of two alternating states: (i) continuous circu-
lation, and (ii) cleaning and dilution. During
continuous circulation, the peristaltic pump
moves the microculture around the growth
loop at a linear velocity of È250 mm sj1 (Fig.
1C). During cleaning and dilution, the mixing
is halted and a segment is isolated from the
rest of the reactor with micromechanical
valves. A lysis buffer is flushed through the
isolated segment for 50 s to expel the cells it
contains, including any wall-adhering cells
(Fig. 1D). Next, the segment is flushed with
sterile growth medium to completely rinse out
the lysis buffer. This segment, filled with sterile
medium, is then reunited with the rest of the
growth chamber, at which point continuous
circulation resumes. This process is repeated
sequentially on different growth chamber seg-
ments, thus eliminating biofilm formation and
enabling pseudocontinuous operation. In com-
parison, passive treatment of the microfluidic
surfaces with nonadhesive surface coatings
Esuch as poly (ethylene glycol), ethylenediami-
netetraacetic acid, polyoxyethylene sorbitan
monolaurate, and bovine serum albumin^
proved ineffective in preventing biofilm forma-
1
Department of Applied Physics, 2
Division of Chem-
istry and Chemical Engineering, California Institute of
Technology, Pasadena, CA 91125, USA.
*Present address: Department of Bioengineering,
Stanford University, Stanford, CA 94305, USA.
.These authors contributed equally to this work.
-Present address: Department of Biomedical Engi-
neering and Institute for Genome Sciences and
Policy, Duke University, Durham, NC 27708, USA.
`Present address: Department of Physics and Astron-
omy, University of British Columbia, Vancouver, BC
V6T 1Z4, Canada.
¬To whom correspondence should be addressed.
E-mail: quake@stanford.edu
R E P O R T S
www.sciencemag.org SCIENCE VOL 309 1 JULY 2005 137
2. tion. Without active removal, biofilms invaded
the fluidic channels within È48 hours (14).
We performed more than 40 growth exper-
iments with Escherichia coli MG1655 cells in
five different chips, using a variety of growth
media ([MOPS EZ Rich (Teknova, Inc.)] and
LB broth with various concentrations of glucose
and bacto-tryptone) at 21-C and 32-C. Upon
inoculation, a typical culture began with a short
lag period, followed by an exponential growth
phase that gave way to a steady-state regime
(Fig. 2A). Steady-state growth was achieved
over a range of dilution rates (0.072 to 0.37
hourj1); wash-out was observed at high dilu-
tion rates. The steady-state cell concentrations
scaled with dilution rate and nutrient richness,
decreasing with increasing dilution rates or de-
creasing bacto-tryptone concentration (Fig. 2B).
To demonstrate the ability of the microche-
mostat to facilitate analysis of complex growth
dynamics, we used it to monitor the dynamics of
cell populations containing a synthetic ‘‘popula-
tion control[ circuit (15), which autonomously
regulates the cell density through a negative
feedback system based on quorum sensing
(16). With the circuit ON, the cell density is
broadcast and detected through the synthesis
and sensing of a signaling molecule (acyl-
homoserine lactone, or AHL), which in turn
modulates the expression of a killer gene (lacZa-
ccdB). The killer gene regulates cell density by
controlling the cell death rate. The circuit, under
control of a synthetic promoter, is inducible with
isopropyl-b-D-thiogalactopyranoside (IPTG) (fig.
S1). The population control circuit_s use of cell-
cell communication enables the programming
of a bacterial population such that the circuit
is insulated from noise in gene expression as
Peristaltic pump
Input
ports
Supply
channels
Growth
chamber
loop Waste
Lysis buffer
Peristaltic pump
Growth
chamber
loop
Medium
Supply
channels
B
C Lysis buffer
N dilution
compartment
Waste
Peristaltic pump
Medium
Supply
channels
D
th
A
Fig. 1. (A) Optical micrograph showing six microchemostats that operate in parallel on a single chip.
Various inputs have been loaded with food dyes to visualize channels and sub-elements of the
microchemostats. The coin is 18 mm in diameter. (B) Optical micrograph showing a single
microchemostat and its main components. Scale bar, 2 mm. (C) Schematic diagram of a microchemostat
in continuous circulation mode. Elements such as the growth loop with individually addressable
connected segments, the peristaltic pump, supply channels, and input/output ports are labeled. (D)
Isolation of a segment from the rest of the growth chamber during cleaning and dilution mode. A lysis
buffer (indicated in red) is introduced into the chip through the lysis buffer port. Integrated microvalves
direct the buffer through the segment, flushing out cells, including those adhering to chamber walls. The
segment is then rinsed with fresh sterile medium and reunited with the rest of the growth chamber.
Fig. 2. (A) Bacterial
growth as a function of
time in various media
and dilution rates D: (1)
MOPS EZ Rich, in 1.1 M
glucose, D 0 0.34 hourj1,
at 32-C; (2) MOPS EZ
Rich, in 0.11 M glucose,
D 0 0.30 hourj1, at
32-C; (3) LB, in 0.5 g/L
of bacto-tryptone, D 0
0.24 hourj1, at 21-C;
(4) LB, in 0.5 g/L of
bacto-tryptone, D 0 0.30
hour–1, at 21-C; (5) LB, in
3 g/L of bacto-tryptone,
D 0 0.37 hourj1, at
21-C; (6) LB, in 0.5 g/L
of bacto-tryptone, D 0
0.37 hourj1, at 21-C;
and (7) LB, in 0.1 g/L of
bacto-tryptone, D 0 0.37
hourj1, at 21-C. The red curves (5, 6, and 7) represent different
concentrations of bacto-tryptone in LB at a fixed dilution rate, and the
open circles (3, 4, and 6) represent constant influent nutrient composition
at various dilution rates. Cultures 3 to 7 were cultivated on a single chip,
whereas cultures 1 and 2 were each cultivated on separate chips. (B)
Graph showing steady-state populations for various choices of dilution
rate and nutrient concentrations. The error bars represent the variation
in the measured steady-state cell density. The steady-state concentra-
tion decreases as the dilution rate increases and increases in proportion
to the influent nutrient richness. Black squares, 0.1 g/L of bacto-
tryptone; red diamonds, 0.5 g/L of bacto-tryptone; blue circles, 3 g/L of
bacto-tryptone.
3
2
1
0
0 25 50 75
1
2
3
4
5
6
7
Hours
Cells/pL
Dilution Rate [1/Hr]
Cells/pL
0.0
0.4
0.8
1.2
1.6
2.0
0.24 0.28 0.32 0.36
A B
R E P O R T S
1 JULY 2005 VOL 309 SCIENCE www.sciencemag.org138
3. well as intercellular phenotypical variability.
This circuit has been characterized in detail
for macroscopic cultures (15), which makes it
particularly appropriate for evaluating per-
formance of the microchemostat in a well-
controlled manner.
A microchemostat chip was used to
perform six simultaneous experiments with
E. coli MC4100Z1 cells and a dilution rate
of 0.16 hour–1 (Fig. 3A). Cultures in reactors
1 to 3 with circuit-bearing cells were induced
with IPTG (circuit ON), whereas those in 5
and 6 were not induced (circuit OFF). Reactor
4 contained a circuit-free population with
IPTG. Circuit-free and circuit-OFF cultures
(4, 5, and 6) grew exponentially to a steady-
state density of È3.5 cells/pL. In contrast,
circuit-ON populations (1, 2, and 3) exhibited
oscillatory dynamics before reaching a lower
steady-state population density after È125
hours. Variations in cell density due to the
discretized nature of the microchemostat
dilution scheme were negligible compared to
the overall population fluctuations (Fig. 3B).
Using the ability to monitor individual cells
in the microchemostat cultures, we observed
that the oscillations in cell density correlated
with specific cell morphologies (Fig. 3A). For
example, upon inoculation, culture 3 (Fig. 3A,
point a) was composed of healthy (small and
cylindrical) cells. With negligible expression
of the killer protein (LacZa-CcdB) at such
low density, the population initially enjoyed
exponential growth, in tandem with the OFF
cultures. The cells were generally healthy dur-
ing this phase, evident in their morphology
(Fig. 3A, point b). However, as the increased
cell density led to increased AHL concentra-
tion and, consequently, increased expression
of the killer protein (Fig. 3A, point c), the cell
density began to decrease. By this time, a frac-
tion of cells had become filamented, showing
the deleterious effect of LacZa-CcdB; due to a
lag in the turnover of the signal (by dilution
and degradation) and that of the killer protein
(by cell division and degradation), cell death
intensified (Fig. 3A, point d), leading to a sharp
decrease in the cell density. Further decreases
in cell density ultimately led to a decrease in
the signal concentration as well as the killer
protein concentration. Eventually, when the
death rate dropped below the growth rate as
the killer protein was diluted out (Fig. 3A,
point e), the population recovered and entered
the next cycle. Culture 3 escaped circuit regu-
lation after 186 hours. Under these conditions,
the bacterial population oscillated for 4 to 6
cycles before approaching a steady-state con-
centration of È2 cells/pL. Cultures 1 and 2 dem-
onstrated similar dynamics.
Different cell morphologies and circuit dy-
namics were apparent when the population con-
trol circuit was introduced into a different E.
coli host strain. In Top10F¶ cells, more com-
plete induction of the circuit was achieved (14),
leading to stronger growth regulation (Fig. 4).
At time 0 in these experiments (with a dilution
rate of 0.16 hour–1), the circuit was turned
ON in cultures 1 to 3 but left OFF in cultures
4 to 6. The OFF cultures (cultures 4 to 6, 0 to
96 hours) grew to a steady-state density of
È3 cells/pL. In contrast, the ON density was
sixfold lower than the OFF density and oscil-
lated about È0.5 cells/pL (1, 0 to 96 hours; 2
and 3, 0 to 44 hours). Top10F¶ cells displayed
none of the morphological responses to circuit
regulation that were evident in MC4100Z1
cells; they always looked small and cylindrical,
similar to circuit-OFF cells.
In general, the circuit appeared to be more
stable in the microchemostats than in macro-
scale batch cultures under otherwise similar
growth conditions. During macroscale experi-
ments with reaction volumes of 3 to 50 ml,
circuit-ON cultures lost regulation within È70
hours for MC4100Z1 and 48 hours for Top10F¶
cells (14). Population control in the microche-
Fig. 3. (A) Growth of
MC4100Z1 cells with
the population-control
circuit ON (reactors 1
to 3), OFF (reactors 5
to 6), or absent (re-
actor 4) on a single
chip. Bottom panels (a
to e) show micro-
graphs of the culture
in reactor 3 at the
corresponding points
during the first oscilla-
tion (scale bar, 25 mm).
Cells were grown at
32-C in LBK medium
(14), buffered at pH
7.6, at a dilution rate
of 0.16 hourj1. The
section enclosed by
the red box is enlarged
in the next panel. (B)
Zoom-in on series 4, 5,
and 6, showing varia-
tion with time in cell
density. The shaded
strips represent dilution
intermissions.
a b c d e
0 25 50 75 100 125 150 175 200
0
1
2
3
4
c
e
db
a
5
6
4
3
2
1
1-3 ON
4 Plasmid-free
5-6 OFF
Cells/pL
Hours
125125 126126 127127
3.23.2
3.43.4
3.63.6
3.83.8
4.04.0
4.24.2
4.44.4
HoursHours
Cells/pLCells/pL
A
B
4
5
6
Fig. 4. Growth of Top10F¶
cells with the circuit cycling
between ON and OFF in the
chip. Initially, cultures 1, 2,
and 3 were ON, while cul-
tures 4, 5, and 6 were OFF.
At 44 hours (point A), cul-
tures 2 and 3 were turned
OFF. At 96 hours (point B),
culture 1 was turned OFF,
and cultures 2 to 6 were
turned ON. Cultures 2 and
3 were cultivated on a sep-
arate chip in a different
experiment under the same conditions. When turned OFF, culture 1 (at 96 hours) and cultures 2 and 3
(at 44 hours) grew exponentially to a density of È3 cells/pL. Upon circuit activation at 96 hours after an
extended OFF period, culture 4 generated sustained oscillations similar to those of culture 1 between 0 and
96 hours, after a rapid decrease in cell density. In comparison, when switched ON at 96 hours, cultures 2, 3,
5, and 6 only briefly demonstrated circuit regulation (evident in the sharp decrease in cell density) before
bouncing back to a high density. Cells were grown at 32-C in LBK medium (14), buffered at pH 7.0.
0 25 50 75 100 125 150 175 200
0
1
2
3
4
BA
6
5
4
3
2
1
Cells/pL
Hours
R E P O R T S
www.sciencemag.org SCIENCE VOL 309 1 JULY 2005 139
4. mostat, in contrast, was routinely maintained for
more than 200 hours and sometimes more than
500 hours (Fig. 5). In theory, the small pop-
ulation size in the microchemostat (È102 to
È104 cells versus È109 cells in macroscale
cultures) should reduce the overall rate at which
mutants may occur and take over a population.
Exactly how the cells lost regulation in the
circuit-ON cultures (Fig. 3, culture 3, and Fig.
4, cultures 2, 3, 5, and 6) is unclear. A simple
mutation rate versus population size argument
would predict much longer lifetimes for main-
taining circuit regulation.
We observed oscillations in cell density for
both E. coli strains in the circuit-ON state. The
MC4100Z1 strain had large-amplitude (È2
cells/pL) oscillations, which gradually decayed
to a steady state (Fig. 3A). The Top10F¶ strain
had smaller amplitude oscillations (È1 cell/pL)
which continued for the length of the measure-
ment (Figs. 4 and 5). Although the origin of
these oscillations is unknown, they may be a
consequence of the circuit interacting with the
continuous culture mechanism of the micro-
chemostat. It is also possible that the oscil-
lations are entirely due to circuit regulation, as
predicted by a simple mathematical model with
biologically feasible parameters (14). These
population-level oscillations are controllable—
they only occur when the circuit is in the ON
state—and they are more sustained and stable
than those generated by synthetic oscillators
operating in individual cells (17, 18).
An active approach to preventing biofilm
formation in microfluidic devices enabled us to
implement a miniaturized bioreactor that oper-
ates at a working volume of 16 nL, more than
300 times smaller than the smallest previous
microfermentor (7). This miniaturized device
enabled automated culturing and monitoring of
populations of È100 to È104 bacteria with
instantaneous single-cell resolution. Reducing
the reactor volume by a factor of 105 can, in
theory, suppress the total mutation rate pro-
portionately and hence prolong monitoring of
genetically homogeneous populations (14).
The microchemostat has enabled us to monitor
the programmed behavior of bacterial popula-
tions for hundreds of hours despite strong
selection pressure to evade population control,
something that was not achieved in macro-
scopic reactors. Although we focused on the
bacterial count and cell morphology, measure-
ments can be readily extended to dynamic
properties, for example, gene expression dy-
namics and distributions reported by fluores-
cence or luminescence. These capabilities
enable long-term, low-cost, high-resolution
investigation of the phenotypical character-
istics of many different cell strains, as well as
natural and synthetic cellular networks under a
matrix of conditions. This capability will
greatly facilitate high-throughput screening
applications in fields such as chemical genetics
and pharmaceutical discovery.
References and Notes
1. J. Monod, Ann. Inst. Pasteur (Paris) 79, 390 (1950).
2. A. Novick, L. Szilard, Science 112, 715 (1950).
3. D. E. Dykhuizen, Annu. Rev. Ecol. Syst. 21, 373 (1990).
4. M. S. Fox, J. Gen. Physiol. 39, 267 (1955).
5. H. L. Smith, P. Waltman, in The Theory of the
Chemostat: Dynamics of Microbial Competition
(Univ. of Cambridge Press, Cambridge, ed. 1, 1995).
6. A. Novick, Annu. Rev. Microbiol. 9, 97 (1955).
7. A. Zanzotto et al., Biotechnol. Bioeng. 87, 243 (2004).
8. J. W. Kim, Y. H. Lee, J. Korean Phys. Soc. 33, S462 (1998).
9. Y. Kostov, P. Harms, L. Randers-Eichhorn, G. Rao,
Biotechnol. Bioeng. 72, 346 (2001).
10. M. M. Maharbiz, W. J. Holtz, R. T. Howe, J. D.
Keasling, Biotechnol. Bioeng. 85, 376 (2004).
11. J. W. Costerton, Z. Lewandowski, D. E. Caldwell, D. R.
Korber, H. M. Lappinscott, Annu. Rev. Microbiol. 49,
711 (1995).
12. H. H. Topiwala, C. Hamer, Biotechnol. Bioeng. 13, 919
(1971).
13. D. H. Larsen, R. L. Dimmick, J. Bacteriol. 88, 1380 (1964).
14. Materials and methods are available as supporting
material on Science Online.
15. L. You, R. S. Cox III, R. Weiss, F. H. Arnold, Nature
428, 868 (2004).
16. M. B. Miller, B. L. Bassler, Annu. Rev. Microbiol. 55,
165 (2001).
17. M. B. Elowitz, S. Leibler, Nature 403, 335 (2000).
18. M. R. Atkinson, M. A. Savageau, J. T. Myers, A. J.
Ninfa, Cell 113, 597 (2003).
19. We thank M. Elowitz for MC4100Z1 cells, U. Alon for
MG1655 cells, T. Ozdere for generating data for figs. S4
and S5, C. Collins for plasmid pLuxR, M. Barnett for tech-
nical assistance, and J. Leadbetter, C. Ward, T. Squires, J.
Huang, and E. Kartalov for helpful discussions. Supported
in part by the NSF and the Defense Advanced Research
Projects Agency (contract no. N66001-02-1-8929).
Supporting Online Material
www.sciencemag.org/cgi/content/full/309/5731/137/
DC1
Materials and Methods
Figs. S1 to S5
Tables S1 and S2
References and Notes
27 December 2004; accepted 10 May 2005
10.1126/science.1109173
tRNA Actively Shuttles Between
the Nucleus and Cytosol in Yeast
Akira Takano,1
Toshiya Endo,1,3,4
Tohru Yoshihisa1,2
*
Previous evidence suggested that transfer RNAs (tRNAs) cross the nuclear
envelope to the cytosol only once after maturing in the nucleus. We now present
evidence for nuclear import of tRNAs in yeast. Several export mutants ac-
cumulate mature tRNAs in the nucleus even in the absence of transcription.
Import requires energy but not the Ran cycle. These results indicate that tRNAs
shuttle between the nucleus and cytosol.
Nuclear-encoded tRNAs are transcribed, pro-
cessed in the nucleus, and exported to the cytosol
to facilitate translation (1). Moreover, a nuclear
pool of mature tRNAs also exists. In Saccha-
romyces cerevisiae, certain mutants defective in
nuclear transport and tRNA processing accu-
mulate mature tRNAs in the nucleus, which
suggests that nuclear mature tRNAs are in-
termediates waiting for tRNA export (2–5).
Before export, aminoacyl-tRNA synthetases
0 100 200 300 400 500
0.0
0.5
1.0
1.5
Dilution Rate [1/Hr]
0.40 0.160.340.300.270.200.160.13
Cells/pL
Hours
Fig. 5. Effects of the dilution rate on population dynamics of Top10F¶ cells with the population-control
circuit ON. At high dilution rates (to 0.27, 0.30, and 0.34 hour–1), both the amplitude and the period of
oscillations diminished. The culture was approaching wash-out at the highest dilution rate (0.40
hourj1). Large oscillations were recovered when a low dilution rate was restored toward the end of
the experiment. Cells were grown at 32-C in LBK medium (14), buffered at pH 7.0.
R E P O R T S
1 JULY 2005 VOL 309 SCIENCE www.sciencemag.org140