Our Genome-Edited Future: the Promise and the ChallengeOECD Environment
This presentation gives some background of genome editing techniques in the broader context. The focus of the Conference is on agriculture to ensure that one specific sector can be addressed in depth. However, the potential applications of genome editing are much broader than just agriculture and
there are a number of topic areas which cannot be covered thoroughly in the limited time available.
Global developments of genome editing in agricultureOECD Environment
This presentation covers the scope of agricultural applications of genome editing by describing the relevance of these techniques to agriculture especially crop plants, farm animals as well as the foods and feeds derived from them.
based on the following paper
"Enhanced resistance to blast fungus in rice (Oryza sativa L.) by
expressing the ribosome-inactivating protein alpha-momorcharin
Qian Qian1, Lin Huang1, Rong Yi, Shuzhen Wang, Yi Ding∗
State Key Laboratory of Hybrid Rice, Department of Genetics, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, People’s Republic
of China
Root genetic research and its application in plant breeding or crop improvementOm Prakash Patidar
UNIVERSITY OF AGRICULTURAL SCIENCES, DHARWAD
DEPARTMENT OF GENETICS AND PLANT BREEDING Master’s seminar-II
Root genetic research and its applications in plant breeding
Speaker: Om Prakash Patidar Date: 20/03/2015 ID No.: PGS13AGR6140 Time: 3:00 PM
Synopsis
Roots play an essential role in the acquisition of water and minerals from soils. Root system architecture (RSA), the spatial configuration of a root system in the soil, is used to describe the shape and structure of root system. Its importance in plant productivity lies in the fact that major soil resources are heterogeneously distributed in the soil, so that the spatial deployment of roots will substantially determine the ability of a plant to secure edaphic resources. Measuring crop root architecture and assaying for changes in function can be challenging, but examples have emerged showing that modifications to roots result in higher yield and increased stress tolerance.1
A marker-assisted back-crossing (MABC) breeding programme was conducted to improve the root morphological traits, and thereby drought tolerance, of the Indian upland rice variety, Kalinga III. The donor parent was Azucena, an upland japonica variety from Philippines. Five segments on different chromosomes were targeted for introgression; four segments carried QTLs for improved root morphological traits and the fifth carried a recessive QTL for aroma. It significantly increased root length under both irrigated and drought stress treatments.2
Alteration of root system architecture improves drought avoidance through the cloning and characterization of DEEPER ROOTING 1 (DRO1), a rice quantitative trait locus controlling root growth angle. Higher expression of DRO1 increases the root growth angle, whereby roots grow in a more downward direction. Introducing DRO1 into a shallow-rooting rice cultivar by backcrossing enabled the resulting line to avoid drought by increasing deep rooting, which maintained high yield performance under drought conditions relative to the recipient cultivar.3
GmEXPB2, A vegetative -expansin gene, clone from a Pi starvation-induced soybean cDNA library. GmEXPB2 was found to be primarily expressed in roots, and was highly induced by Pi starvation, and the induction pattern was confirmed by GUS staining in transgenic soybean hairy roots. Results from intact soybean composite showed that GmEXPB2 is involved in hairy root elongation, and subsequently affects plant growth and P uptake, especially at low P levels.4
Candidate Aluminum tolerance proteins include organic acid efflux transporters, with the organic acids forming non-toxic complexes with rhizosphere aluminum. ge
Crop plants with improved culture and quality traits for food, feed and othe...OECD Environment
This document summarizes the work of a consortium using genome editing techniques to improve traits in crop plants. Key points:
- The consortium involved 10 public and 4 private labs working on 9 crop species, 3 vegetables, 1 fruit, 1 forestry species, and 3 model plants.
- Successful genome editing was achieved for targeted mutagenesis in all species, while more complex genome editing was successful in only 2 of the 12 species tested.
- Traits improved include increased transformation rates, flowering time, starch composition, stress tolerance, and disease resistance.
- Over 22,000 plants were regenerated in experiments testing new gene editing tools and techniques across species. Genome editing was shown to be an
This document discusses the biosafety of genetically modified crops. It outlines the approach taken to assess safety, including potential risks like toxicity, allergenicity, antibiotic resistance, and gene flow. The regulatory framework for genetically modified crops in India is also mentioned. Specific concerns discussed include Brazil nut allergy in soybean, use of antibiotic resistance marker genes, consumption of foreign DNA, and effects on biodiversity and target species. Strategies to prevent unwanted gene flow are described.
The document discusses several topics related to climate change and plant breeding:
1. It outlines natural and human causes of climate change such as changes in the sun's energy output and increasing greenhouse gas emissions.
2. It describes how plants may respond to climate change through extinction, range shifts, habitat fragmentation, genetic differentiation, migration, and phenotypic plasticity.
3. It discusses strategies for improving crop resilience through crop diversification, ideotype breeding, and mutation breeding. Crop diversification involves mixing varieties to reduce pest and disease impacts. Ideotype breeding develops optimized crop models. Mutation breeding enhances genetic variability.
Application of genome editing in farm animals: Cattle - Alison Van EenennaamOECD Environment
This document summarizes an expert presentation on animal genomics and biotechnology education. It discusses:
1) Cattle contribute significantly to global animal protein supply and demand for cattle products is projected to increase substantially by 2050. Accelerating genetic gain through breeding programs is needed to meet this demand more sustainably.
2) Genome editing holds promise for introducing beneficial traits into cattle, such as polledness, heat tolerance, and disease resistance. One example discussed was using TALENs to introduce the polled allele into dairy cattle to eliminate painful horn removal.
3) However, regulatory hurdles like the FDA's stance that gene-edited animals are drugs could slow the application of new gene editing
Our Genome-Edited Future: the Promise and the ChallengeOECD Environment
This presentation gives some background of genome editing techniques in the broader context. The focus of the Conference is on agriculture to ensure that one specific sector can be addressed in depth. However, the potential applications of genome editing are much broader than just agriculture and
there are a number of topic areas which cannot be covered thoroughly in the limited time available.
Global developments of genome editing in agricultureOECD Environment
This presentation covers the scope of agricultural applications of genome editing by describing the relevance of these techniques to agriculture especially crop plants, farm animals as well as the foods and feeds derived from them.
based on the following paper
"Enhanced resistance to blast fungus in rice (Oryza sativa L.) by
expressing the ribosome-inactivating protein alpha-momorcharin
Qian Qian1, Lin Huang1, Rong Yi, Shuzhen Wang, Yi Ding∗
State Key Laboratory of Hybrid Rice, Department of Genetics, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, People’s Republic
of China
Root genetic research and its application in plant breeding or crop improvementOm Prakash Patidar
UNIVERSITY OF AGRICULTURAL SCIENCES, DHARWAD
DEPARTMENT OF GENETICS AND PLANT BREEDING Master’s seminar-II
Root genetic research and its applications in plant breeding
Speaker: Om Prakash Patidar Date: 20/03/2015 ID No.: PGS13AGR6140 Time: 3:00 PM
Synopsis
Roots play an essential role in the acquisition of water and minerals from soils. Root system architecture (RSA), the spatial configuration of a root system in the soil, is used to describe the shape and structure of root system. Its importance in plant productivity lies in the fact that major soil resources are heterogeneously distributed in the soil, so that the spatial deployment of roots will substantially determine the ability of a plant to secure edaphic resources. Measuring crop root architecture and assaying for changes in function can be challenging, but examples have emerged showing that modifications to roots result in higher yield and increased stress tolerance.1
A marker-assisted back-crossing (MABC) breeding programme was conducted to improve the root morphological traits, and thereby drought tolerance, of the Indian upland rice variety, Kalinga III. The donor parent was Azucena, an upland japonica variety from Philippines. Five segments on different chromosomes were targeted for introgression; four segments carried QTLs for improved root morphological traits and the fifth carried a recessive QTL for aroma. It significantly increased root length under both irrigated and drought stress treatments.2
Alteration of root system architecture improves drought avoidance through the cloning and characterization of DEEPER ROOTING 1 (DRO1), a rice quantitative trait locus controlling root growth angle. Higher expression of DRO1 increases the root growth angle, whereby roots grow in a more downward direction. Introducing DRO1 into a shallow-rooting rice cultivar by backcrossing enabled the resulting line to avoid drought by increasing deep rooting, which maintained high yield performance under drought conditions relative to the recipient cultivar.3
GmEXPB2, A vegetative -expansin gene, clone from a Pi starvation-induced soybean cDNA library. GmEXPB2 was found to be primarily expressed in roots, and was highly induced by Pi starvation, and the induction pattern was confirmed by GUS staining in transgenic soybean hairy roots. Results from intact soybean composite showed that GmEXPB2 is involved in hairy root elongation, and subsequently affects plant growth and P uptake, especially at low P levels.4
Candidate Aluminum tolerance proteins include organic acid efflux transporters, with the organic acids forming non-toxic complexes with rhizosphere aluminum. ge
Crop plants with improved culture and quality traits for food, feed and othe...OECD Environment
This document summarizes the work of a consortium using genome editing techniques to improve traits in crop plants. Key points:
- The consortium involved 10 public and 4 private labs working on 9 crop species, 3 vegetables, 1 fruit, 1 forestry species, and 3 model plants.
- Successful genome editing was achieved for targeted mutagenesis in all species, while more complex genome editing was successful in only 2 of the 12 species tested.
- Traits improved include increased transformation rates, flowering time, starch composition, stress tolerance, and disease resistance.
- Over 22,000 plants were regenerated in experiments testing new gene editing tools and techniques across species. Genome editing was shown to be an
This document discusses the biosafety of genetically modified crops. It outlines the approach taken to assess safety, including potential risks like toxicity, allergenicity, antibiotic resistance, and gene flow. The regulatory framework for genetically modified crops in India is also mentioned. Specific concerns discussed include Brazil nut allergy in soybean, use of antibiotic resistance marker genes, consumption of foreign DNA, and effects on biodiversity and target species. Strategies to prevent unwanted gene flow are described.
The document discusses several topics related to climate change and plant breeding:
1. It outlines natural and human causes of climate change such as changes in the sun's energy output and increasing greenhouse gas emissions.
2. It describes how plants may respond to climate change through extinction, range shifts, habitat fragmentation, genetic differentiation, migration, and phenotypic plasticity.
3. It discusses strategies for improving crop resilience through crop diversification, ideotype breeding, and mutation breeding. Crop diversification involves mixing varieties to reduce pest and disease impacts. Ideotype breeding develops optimized crop models. Mutation breeding enhances genetic variability.
Application of genome editing in farm animals: Cattle - Alison Van EenennaamOECD Environment
This document summarizes an expert presentation on animal genomics and biotechnology education. It discusses:
1) Cattle contribute significantly to global animal protein supply and demand for cattle products is projected to increase substantially by 2050. Accelerating genetic gain through breeding programs is needed to meet this demand more sustainably.
2) Genome editing holds promise for introducing beneficial traits into cattle, such as polledness, heat tolerance, and disease resistance. One example discussed was using TALENs to introduce the polled allele into dairy cattle to eliminate painful horn removal.
3) However, regulatory hurdles like the FDA's stance that gene-edited animals are drugs could slow the application of new gene editing
This document provides an overview of transgenic crops or genetically modified organisms (GMOs). It begins by defining transgenic crops as plants containing genes artificially inserted from unrelated species using recombinant DNA technology. The document then discusses the aims of genetic engineering in crops, including introducing traits like pest and disease resistance. It also summarizes the status and adoption of transgenic crops globally. The rest of the document addresses various myths and controversies around transgenic crops, providing facts and evidence to counter claims about risks to health, the environment and farmers. It concludes by noting both benefits and tensions around GMO technology but argues that advances which contribute to sustainable food security should be welcomed.
Transgenic crops are genetically modified crops containing genes artificially inserted from another species. The first GM crop was a tobacco plant in 1982, and the first approved for sale in the US was the FlavrSavr tomato in 1994. GM crops are developed using genetic engineering techniques to speed up traditional breeding and introduce a wider variety of genes. Potential benefits include increased yields, insect and disease resistance, and improved nutrition. However, there are also concerns about the impacts on human and environmental health.
The document summarizes the principles and practice of environmental safety assessment for transgenic plants. It discusses evaluating the stability of genetic modifications, potential for gene transfer to related and unrelated organisms, weediness potential, and effects on non-target organisms. It provides details on Monsanto's MON 810 Bt corn, including the genetic elements inserted, molecular characterization of the inserted DNA, expression of the Cry1Ab protein, and laboratory and field studies finding no adverse effects on non-target and beneficial organisms except certain lepidopteran pests. It emphasizes the importance of insect resistance management plans to prolong the effectiveness of Bt toxins in crops.
Ethical and bio-safety issues related to GM cropsMahammed Faizan
a seminar presentation on ethical and bio-safety issues related GM crops.
impact of gm crops on human, animal and environmental health.
safety measure related transgenic crops.
international governmental bodies
This document discusses biosafety issues related to genetically modified crops. It provides background on GM crops and their history. It then outlines several biosafety concerns including the safety of inserted genes and proteins, ecological impacts such as increased weediness and effects on biodiversity, environmental concerns like secondary pest problems and insect resistance, and socioeconomic issues. The regulatory mechanisms in place in India to evaluate GM crops are also described, including the various competent authorities. International regulations like the Cartagena Protocol are also mentioned.
Assesment Food and Environmental Saftey of Genetically Engineered (GE) Food C...Suresh Antre
Over the last two decades, individual governments and intergovernmental organizations have designed strategies and protocols for safety assessment of foods/ feed derived from GM crops, (FAO/WHO, CAC, OECD etc).
All GM crops that have been approved for commercialization and entered the agri-food chain have undergone extensive testing.
This document provides information about genetically modified crops. It discusses how GM crops are produced through genetic engineering by inserting genes from other organisms. It then discusses GM crops that have been approved for commercial production, including insect-resistant cotton, maize, and soybean as well as herbicide-tolerant soybean. The document also discusses the global area planted with biotech crops annually and countries that grow the most biotech crops. It outlines some pros and cons of GM crops and how they are regulated in different countries including the US, EU, and India.
This document summarizes significant progress in crop biotechnology. It discusses the history of crop biotechnology from 1983 to present, highlighting major developments such as the first GM plant in 1983 and approval of GM foods by FDA in 1992. The document also covers global area of biotech crops from 1996 to 2016, benefits of GM crops including insect resistance, herbicide tolerance, disease resistance, and improved nutrition. Additionally, it discusses potential risks of GM crops relating to health, environment, and social issues.
The age of gene editing - Workshop on innovations in food and agriculture sys...OECD Environment
The workshop took place in Paris on 25-26 February 2016. Its central aim was to discuss with experts how scientific, technological, and farm practice innovation can improve productivity and sustainability in the food and agricultural sector, with a focus on international collaboration on gene editing techniques. It was introduced in the form of a presentation entitled ‘The Age of Gene editing’, produced by Steffi Friedrichs (STI), which played a pivotal role during the expert discussions.
Golden rice-and-bt-crops-los-banos-phil-08-24-2011Heba FromAlla
Golden Rice and Bt crops: Unanswered safety and efficacy questions
This document outlines several unanswered questions regarding the safety and efficacy of Golden Rice and Bt crops. For Golden Rice, questions remain about how much beta-carotene is retained after storage and cooking. Human feeding trials were canceled after being notified by Greenpeace. There are also environmental and agricultural questions that remain unanswered. For Bt crops, proteomics studies found unintended changes in protein expression levels in Bt maize. Mouse feeding studies also found immune and reproductive system impacts from Bt maize consumption. Carefully designed long-term feeding studies in mice found statistically significant adverse reproductive effects from Bt maize consumption.
This document provides an overview of transgenic crops, including:
1) A brief history of transgenic crop development and the governing policies surrounding the technology.
2) A summary of the main agricultural crops that have been genetically modified, their expressed characteristics, and their market roles.
3) A discussion of unintended consequences, economic considerations, safety concerns, and implications of transgenic crops for sustainable agriculture.
CD Genomics is dedicated to providing a comprehensive list of genomics and microarray solutions for agriculture, including genome, exome, transcriptome, and metagenome sequencing, genome-wide association studies (GWAS), and targeted sequencing and genotyping that focus on a subset of regions or genes such as single nucleotide polymorphisms (SNPs). https://www.cd-genomics.com/Transcriptomics.html
Luis Augusto Becerra presents an overview of Flagship Project 1 'Enhanced genetic resources' of the CGIAR Research Program on Roots, Tubers and Bananas (RTB), during the 18th Triennial Symposium of the International Society of Tropical Roots Crops (ISTRC) in October 2018.
Genetically modified foods are derived from organisms that have had their DNA altered through genetic engineering techniques, allowing genes to be inserted from different species. The first commercial GMO was the Flavr Savr tomato, created by Calgene company in 1994 to have a delayed ripening process. While some GMOs offer benefits like disease resistance or increased nutrition, critics argue they pose risks such as increased cancer risk, allergies, or lower nutritional value.
This document discusses the AB QTL mapping strategy and its applications in various crops. AB QTL mapping involves introgressing genomic regions from unadapted germplasm into elite varieties while performing QTL analysis in advanced backcross generations. The document summarizes AB QTL studies in tomato, rice, maize, and their findings. It notes the advantages of AB QTL over conventional QTL mapping, such as reduced linkage drag and ability to rapidly develop candidate varieties. The document also outlines some limitations of the AB QTL approach.
Next-generation waxy corn – a flagship case of SDN-1/NHEJ genome editing via ...OECD Environment
1) DowDuPont is developing waxy corn varieties using CRISPR/Cas9 genome editing to introduce a targeted mutation in the waxy gene that results in the entire coding sequence being deleted.
2) Trials show the CRISPR-edited waxy corn has identical phenotypes and starch properties as conventional waxy corn varieties.
3) CRISPR-edited waxy corn hybrids have shown improved yields compared to conventional hybrid checks in agronomic trials.
JGI: Genome size impacts on plant adaptationjrossibarra
Genome size may impact how plant genomes adapt, offering larger mutational targets leading to more adaptation from standing variation and more adaptation in noncoding regions.
This document discusses genetically modified crops and their potential environmental impacts. It describes current and future GM crop traits such as insect resistance, herbicide tolerance, stress tolerance, and production of pharmaceuticals in plants. Major concerns discussed include effects on non-target species, gene flow and transgene escape. The document outlines EPA research on monitoring these impacts, including assessing non-target effects using molecular techniques to detect gene expression changes in species exposed to GM crops. It provides examples of monitoring bentgrass for transgene escape and using sentinel plants and resident populations to track gene flow. The overall goals are to apply molecular monitoring to at-risk species and ensure the safety of biotech crops.
Autophagy and its role in plants - By Tilak I S, Dept. of Biotechnology, UASD.Tilak I S
Autophagy (Macroautophagy) a term from the Greek ‘auto’ (self) and ‘phagein’ (to eat), is a highly regulated cellular degradation and recycling process, conserved from yeast to more complex eukaryotes. The process involves sequestration of the cytoplasm into double-membrane vesicles called autophagosomes, which subsequently fuse with lysosomes or vacuoles. The products of autophagic degradation of intracellular material are exported from lysosomes into the cytoplasm where they are recycled (Tang et al., 2018).
Autophagy is activated during various extracellular or intracellular factors such as nutrients deprivation, drought, stresses, and pathogenic invasion to degrade damaged, denatured, and aggregated proteins (Floyd et al., 2015). The mechanism of autophagy induction and regulation is carried out by TOR (Target of Rapamycin) complex and a number of autophagy related genes (ATGs) and proteins which have been identified in higher eukaryotes including yeasts, mammals, and plants (arabidopsis, rice, wheat, tomato and maize etc.) (Ryabovol and Minibayeva., 2016). In plants autophagy is essential for various physiological processes like growth and development, elimination of toxic compounds from the plants Eg: ROS (reactive oxygen species), involved in programmed cell death, nutrients recycling under detrimental environmental factors. Li et al. (2015) transferred an autophagy-related gene, SiATG8a, from foxtail millet to arabidopsis. Through expression profile analyses demonstrated that SiATG8a expression was induced by both drought and nitrogen starvation and over-expression of SiATG8a improved tolerance to nitrogen starvation and drought stress in transgenic Arabidopsis.
The study of autophagy in crop species has been expanding rapidly. Functions of autophagy in development, abiotic stress responses and plant–microbe interactions have been deciphered in various species (Kabbage et al., 2013). New findings such as the involvement of autophagy in reproductive development are increasing our understanding of autophagy but much work is still needed. One interesting topic that warrants more attention is the role of autophagy in organs or tissues that are specifically present in certain crops, for example fruits and nodules.
Considering its importance in development and stress responses, autophagy is a promising target to manipulate for agricultural benefits like higher yield. Increased expression of ATG genes may be valuable in agricultural applications, as this can confer a number of benefits to plants, including enhanced growth, higher yield and increased stress tolerance.
Do plants contain typical GPCRs?” How is G-protein signaling operating in plants.
G-proteins are universal signal transducers mediating many cellular responses. In animal systems the G-protein signaling cycle is activated by seven transmembrane-spanning G-protein coupled receptors (or GPCRs, popularly known as “serpentine receptors”). Whether typical G protein-coupled receptors (GPCRs) exist in plants or not is a fundamental question. In contrast to the animal system, the existence of these types of receptors in plants still remains controversial. While in animals ligand binding causes a change in receptor conformation that activate a particular G Protein, in plants, such mechanism is unknown. In fact, it is considered that the plants G-Proteins are self-activating. The G Proteins have their respective GPCRs in animal system. A lot of information is already accumulated in animal system and hence the animal GPCRs are considered “canonical.” Thus, from the very beginning, plant G-proteins have been compared with the animal counterparts and studied as an extrapolation of the animal model. This presentation provides an insight into the molecular mechanisms of G Protein activation in plants as well as whether “canonical” GPCRs are present in any plant species or not.
This document provides an overview of transgenic crops or genetically modified organisms (GMOs). It begins by defining transgenic crops as plants containing genes artificially inserted from unrelated species using recombinant DNA technology. The document then discusses the aims of genetic engineering in crops, including introducing traits like pest and disease resistance. It also summarizes the status and adoption of transgenic crops globally. The rest of the document addresses various myths and controversies around transgenic crops, providing facts and evidence to counter claims about risks to health, the environment and farmers. It concludes by noting both benefits and tensions around GMO technology but argues that advances which contribute to sustainable food security should be welcomed.
Transgenic crops are genetically modified crops containing genes artificially inserted from another species. The first GM crop was a tobacco plant in 1982, and the first approved for sale in the US was the FlavrSavr tomato in 1994. GM crops are developed using genetic engineering techniques to speed up traditional breeding and introduce a wider variety of genes. Potential benefits include increased yields, insect and disease resistance, and improved nutrition. However, there are also concerns about the impacts on human and environmental health.
The document summarizes the principles and practice of environmental safety assessment for transgenic plants. It discusses evaluating the stability of genetic modifications, potential for gene transfer to related and unrelated organisms, weediness potential, and effects on non-target organisms. It provides details on Monsanto's MON 810 Bt corn, including the genetic elements inserted, molecular characterization of the inserted DNA, expression of the Cry1Ab protein, and laboratory and field studies finding no adverse effects on non-target and beneficial organisms except certain lepidopteran pests. It emphasizes the importance of insect resistance management plans to prolong the effectiveness of Bt toxins in crops.
Ethical and bio-safety issues related to GM cropsMahammed Faizan
a seminar presentation on ethical and bio-safety issues related GM crops.
impact of gm crops on human, animal and environmental health.
safety measure related transgenic crops.
international governmental bodies
This document discusses biosafety issues related to genetically modified crops. It provides background on GM crops and their history. It then outlines several biosafety concerns including the safety of inserted genes and proteins, ecological impacts such as increased weediness and effects on biodiversity, environmental concerns like secondary pest problems and insect resistance, and socioeconomic issues. The regulatory mechanisms in place in India to evaluate GM crops are also described, including the various competent authorities. International regulations like the Cartagena Protocol are also mentioned.
Assesment Food and Environmental Saftey of Genetically Engineered (GE) Food C...Suresh Antre
Over the last two decades, individual governments and intergovernmental organizations have designed strategies and protocols for safety assessment of foods/ feed derived from GM crops, (FAO/WHO, CAC, OECD etc).
All GM crops that have been approved for commercialization and entered the agri-food chain have undergone extensive testing.
This document provides information about genetically modified crops. It discusses how GM crops are produced through genetic engineering by inserting genes from other organisms. It then discusses GM crops that have been approved for commercial production, including insect-resistant cotton, maize, and soybean as well as herbicide-tolerant soybean. The document also discusses the global area planted with biotech crops annually and countries that grow the most biotech crops. It outlines some pros and cons of GM crops and how they are regulated in different countries including the US, EU, and India.
This document summarizes significant progress in crop biotechnology. It discusses the history of crop biotechnology from 1983 to present, highlighting major developments such as the first GM plant in 1983 and approval of GM foods by FDA in 1992. The document also covers global area of biotech crops from 1996 to 2016, benefits of GM crops including insect resistance, herbicide tolerance, disease resistance, and improved nutrition. Additionally, it discusses potential risks of GM crops relating to health, environment, and social issues.
The age of gene editing - Workshop on innovations in food and agriculture sys...OECD Environment
The workshop took place in Paris on 25-26 February 2016. Its central aim was to discuss with experts how scientific, technological, and farm practice innovation can improve productivity and sustainability in the food and agricultural sector, with a focus on international collaboration on gene editing techniques. It was introduced in the form of a presentation entitled ‘The Age of Gene editing’, produced by Steffi Friedrichs (STI), which played a pivotal role during the expert discussions.
Golden rice-and-bt-crops-los-banos-phil-08-24-2011Heba FromAlla
Golden Rice and Bt crops: Unanswered safety and efficacy questions
This document outlines several unanswered questions regarding the safety and efficacy of Golden Rice and Bt crops. For Golden Rice, questions remain about how much beta-carotene is retained after storage and cooking. Human feeding trials were canceled after being notified by Greenpeace. There are also environmental and agricultural questions that remain unanswered. For Bt crops, proteomics studies found unintended changes in protein expression levels in Bt maize. Mouse feeding studies also found immune and reproductive system impacts from Bt maize consumption. Carefully designed long-term feeding studies in mice found statistically significant adverse reproductive effects from Bt maize consumption.
This document provides an overview of transgenic crops, including:
1) A brief history of transgenic crop development and the governing policies surrounding the technology.
2) A summary of the main agricultural crops that have been genetically modified, their expressed characteristics, and their market roles.
3) A discussion of unintended consequences, economic considerations, safety concerns, and implications of transgenic crops for sustainable agriculture.
CD Genomics is dedicated to providing a comprehensive list of genomics and microarray solutions for agriculture, including genome, exome, transcriptome, and metagenome sequencing, genome-wide association studies (GWAS), and targeted sequencing and genotyping that focus on a subset of regions or genes such as single nucleotide polymorphisms (SNPs). https://www.cd-genomics.com/Transcriptomics.html
Luis Augusto Becerra presents an overview of Flagship Project 1 'Enhanced genetic resources' of the CGIAR Research Program on Roots, Tubers and Bananas (RTB), during the 18th Triennial Symposium of the International Society of Tropical Roots Crops (ISTRC) in October 2018.
Genetically modified foods are derived from organisms that have had their DNA altered through genetic engineering techniques, allowing genes to be inserted from different species. The first commercial GMO was the Flavr Savr tomato, created by Calgene company in 1994 to have a delayed ripening process. While some GMOs offer benefits like disease resistance or increased nutrition, critics argue they pose risks such as increased cancer risk, allergies, or lower nutritional value.
This document discusses the AB QTL mapping strategy and its applications in various crops. AB QTL mapping involves introgressing genomic regions from unadapted germplasm into elite varieties while performing QTL analysis in advanced backcross generations. The document summarizes AB QTL studies in tomato, rice, maize, and their findings. It notes the advantages of AB QTL over conventional QTL mapping, such as reduced linkage drag and ability to rapidly develop candidate varieties. The document also outlines some limitations of the AB QTL approach.
Next-generation waxy corn – a flagship case of SDN-1/NHEJ genome editing via ...OECD Environment
1) DowDuPont is developing waxy corn varieties using CRISPR/Cas9 genome editing to introduce a targeted mutation in the waxy gene that results in the entire coding sequence being deleted.
2) Trials show the CRISPR-edited waxy corn has identical phenotypes and starch properties as conventional waxy corn varieties.
3) CRISPR-edited waxy corn hybrids have shown improved yields compared to conventional hybrid checks in agronomic trials.
JGI: Genome size impacts on plant adaptationjrossibarra
Genome size may impact how plant genomes adapt, offering larger mutational targets leading to more adaptation from standing variation and more adaptation in noncoding regions.
This document discusses genetically modified crops and their potential environmental impacts. It describes current and future GM crop traits such as insect resistance, herbicide tolerance, stress tolerance, and production of pharmaceuticals in plants. Major concerns discussed include effects on non-target species, gene flow and transgene escape. The document outlines EPA research on monitoring these impacts, including assessing non-target effects using molecular techniques to detect gene expression changes in species exposed to GM crops. It provides examples of monitoring bentgrass for transgene escape and using sentinel plants and resident populations to track gene flow. The overall goals are to apply molecular monitoring to at-risk species and ensure the safety of biotech crops.
Autophagy and its role in plants - By Tilak I S, Dept. of Biotechnology, UASD.Tilak I S
Autophagy (Macroautophagy) a term from the Greek ‘auto’ (self) and ‘phagein’ (to eat), is a highly regulated cellular degradation and recycling process, conserved from yeast to more complex eukaryotes. The process involves sequestration of the cytoplasm into double-membrane vesicles called autophagosomes, which subsequently fuse with lysosomes or vacuoles. The products of autophagic degradation of intracellular material are exported from lysosomes into the cytoplasm where they are recycled (Tang et al., 2018).
Autophagy is activated during various extracellular or intracellular factors such as nutrients deprivation, drought, stresses, and pathogenic invasion to degrade damaged, denatured, and aggregated proteins (Floyd et al., 2015). The mechanism of autophagy induction and regulation is carried out by TOR (Target of Rapamycin) complex and a number of autophagy related genes (ATGs) and proteins which have been identified in higher eukaryotes including yeasts, mammals, and plants (arabidopsis, rice, wheat, tomato and maize etc.) (Ryabovol and Minibayeva., 2016). In plants autophagy is essential for various physiological processes like growth and development, elimination of toxic compounds from the plants Eg: ROS (reactive oxygen species), involved in programmed cell death, nutrients recycling under detrimental environmental factors. Li et al. (2015) transferred an autophagy-related gene, SiATG8a, from foxtail millet to arabidopsis. Through expression profile analyses demonstrated that SiATG8a expression was induced by both drought and nitrogen starvation and over-expression of SiATG8a improved tolerance to nitrogen starvation and drought stress in transgenic Arabidopsis.
The study of autophagy in crop species has been expanding rapidly. Functions of autophagy in development, abiotic stress responses and plant–microbe interactions have been deciphered in various species (Kabbage et al., 2013). New findings such as the involvement of autophagy in reproductive development are increasing our understanding of autophagy but much work is still needed. One interesting topic that warrants more attention is the role of autophagy in organs or tissues that are specifically present in certain crops, for example fruits and nodules.
Considering its importance in development and stress responses, autophagy is a promising target to manipulate for agricultural benefits like higher yield. Increased expression of ATG genes may be valuable in agricultural applications, as this can confer a number of benefits to plants, including enhanced growth, higher yield and increased stress tolerance.
Do plants contain typical GPCRs?” How is G-protein signaling operating in plants.
G-proteins are universal signal transducers mediating many cellular responses. In animal systems the G-protein signaling cycle is activated by seven transmembrane-spanning G-protein coupled receptors (or GPCRs, popularly known as “serpentine receptors”). Whether typical G protein-coupled receptors (GPCRs) exist in plants or not is a fundamental question. In contrast to the animal system, the existence of these types of receptors in plants still remains controversial. While in animals ligand binding causes a change in receptor conformation that activate a particular G Protein, in plants, such mechanism is unknown. In fact, it is considered that the plants G-Proteins are self-activating. The G Proteins have their respective GPCRs in animal system. A lot of information is already accumulated in animal system and hence the animal GPCRs are considered “canonical.” Thus, from the very beginning, plant G-proteins have been compared with the animal counterparts and studied as an extrapolation of the animal model. This presentation provides an insight into the molecular mechanisms of G Protein activation in plants as well as whether “canonical” GPCRs are present in any plant species or not.
This document summarizes heterotrimeric G-proteins. It discusses that G-proteins are guanine nucleotide binding proteins composed of three subunits - alpha, beta, and gamma. The alpha subunit acts as a molecular switch cycling between an active GTP-bound form and inactive GDP-bound form. When a receptor is activated by a ligand, it causes a conformational change in the G-protein alpha subunit, activating it to turn on downstream effector molecules. The mechanism and roles of each subunit are described. Examples are given of how cholera toxin can cause disease by modifying G-protein alpha subunits and deregulating ion transport.
ARGOS8 variants generated by CRISPR-Cas9 improve maize grain yield under field...Ahmed Madni
Maize ARGOS8 is a negative regulator of ethylene responses. A previous study has shown that
transgenic plants constitutively overexpressing ARGOS8 have reduced ethylene sensitivity and
improved grain yield under drought stress conditions. To explore the targeted use of ARGOS8
native expression variation in drought-tolerant breeding, a diverse set of over 400 maize inbreds
was examined for ARGOS8 mRNA expression, but the expression levels in all lines were less than
that created in the original ARGOS8 transgenic events. We then employed a CRISPR-Cas-enabled
advanced breeding technology to generate novel variants of ARGOS8. The native maize GOS2
promoter, which confers a moderate level of constitutive expression, was inserted into the
50-untranslated region of the native ARGOS8 gene or was used to replace the native promoter of
ARGOS8. Precise genomic DNA modification at the ARGOS8 locus was verified by PCR and
sequencing. The ARGOS8 variants had elevated levels of ARGOS8 transcripts relative to the
native allele and these transcripts were detectable in all the tissues tested, which was the
expected results using the GOS2 promoter. A field study showed that compared to the WT, the
ARGOS8 variants increased grain yield by five bushels per acre under flowering stress conditions
and had no yield loss under well-watered conditions. These results demonstrate the utility of the
CRISPR-Cas9 system in generating novel allelic variation for breeding drought-tolerant crops.
This document discusses biofortification of vegetable crops to combat hidden hunger. It defines biofortification as increasing micronutrients in edible parts of crops through breeding. Methods include agronomic, conventional, and genetic engineering approaches. Case studies show biofortifying crops like cassava, potato, and sweet potato to increase carotenoids, iron, zinc and protein through breeding. Rapid cycling selection in cassava reduced time to improve carotenoids. Co-localizing QTL for iron and zinc in common bean allowed improving both simultaneously. Overall, biofortification is a promising strategy to provide micronutrients and combat malnutrition in a sustainable way.
JBEI Research Highlights - September 2018 Irina Silva
This document summarizes research on characterizing the activities of lignin-modifying enzymes using nanostructure-initiator mass spectrometry (NIMS). NIMS was used to rapidly analyze product formation and kinetics from reactions of a laccase and manganese peroxidase with phenolic and non-phenolic β-aryl ether model lignin substrates. Different primary reaction pathways were observed for each enzyme due to availability of phenoxy radical intermediates. NIMS provided quantitative analysis of bond cleavage events not available from conventional assays and can be performed in microliter volumes.
JBEI Research Highlights - September 2018 Irina Silva
- Three members of the Arabidopsis thaliana glycosyltransferase family 92 (GALS1, GALS2, GALS3) were found to be β-1,4-galactan synthases that add galactose residues to the rhamnogalacturonan-I backbone.
- Overexpression of GALS proteins led to accumulation of unbranched β-1,4-galactan in Arabidopsis, while triple knockout mutants lacked detectable β-1,4-galactan but showed no developmental phenotypes.
- The study provides insight into the properties and roles of these galactan synthase enzymes in pectin biosynthesis in Arabidopsis.
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1) Mga1 was cloned from M. ruber and shown to regulate growth, development and secondary metabolite production.
2) Deletion of Mga1 resulted in reduced growth, defects in hyphal morphology and loss of pigment production.
3) Mga1 deletion mutants showed temperature sensitivity and exogenous cAMP partially restored growth, suggesting Mga1 signals through a cAMP/PKA pathway.
4) Mga1 is required for sexual reproduction in M. ruber.
Omics related approaches for higher productivity and improved quality.pptxAnirudhTV
The document discusses using omics approaches to improve crop productivity and quality. It covers various omics fields including genomics, epigenomics, transcriptomics, proteomics, metabolomics, and phenomics. Examples are provided on applying these approaches in crops like rice, tomato, groundnut, and brassica to traits such as drought tolerance, nutrient enrichment, and reduced anti-nutrients. A case study on analyzing protein abundance changes in wheat cultivars under drought stress using proteomics is also mentioned.
This document summarizes information about two herbicide resistance genes - bar and pat genes. The bar gene codes for the enzyme phosphinothricin acetyl transferase (PAT) which inactivates the herbicide phosphinothricin (PPT). Plants containing the bar gene are tolerant to PPT. The bar gene has been introduced into crops like rapeseed oil and maize. The document also discusses imidazolinone herbicides and tolerance achieved by mutating the acetolactate synthase gene in maize. Environmental impacts of herbicide tolerant crops include concerns about reducing biodiversity and development of herbicide resistant weeds.
High-value pleiotropic genes for developing multiple stress-tolerant biofort...PABOLU TEJASREE
Modern agriculture confronts multifaceted challenges, encompassing biotic and abiotic stresses alongside malnutrition. Biofortified crops emerge as a pivotal solution, augmenting nutritional quality during plant growth. By harnessing specific genes with pleiotropic effects for stress tolerance, these crops exhibit heightened yields, resilience against pests and diseases, and adaptability to environmental stressors. This innovation not only secures food safety and nutrition but also fosters the development of "high-value farms," ensuring sustainable escalation in global food productivity and stable food prices.
Conclusion: Integrating diverse transgenes and gene editing with omics approaches enhances stress tolerance and nutritional content in biofortified crops. This holistic strategy enables precise modifications to crop genomes and comprehensive insights into stress responses and nutrient metabolism, ensuring sustainable food production and nutrition security.
Prospectus and issues of transgenics in agricultureSachin Ekatpure
This document provides an overview of prospects and issues related to transgenic crops. It defines what a transgenic is and describes the process of producing transgenic plants using recombinant DNA technology. It discusses various applications of transgenic crops like herbicide tolerance, insect resistance, virus resistance, and improved nutrition. It also outlines regulatory frameworks for biosafety and examines potential risks like toxicity, gene flow, development of resistance, and impact on biodiversity. The document concludes by noting strategies to minimize risks and future prospects of transgenic technology.
Functional Analysis Of Heterologous Gpcr Signaling Pathways In Yeastbeneshjoseph
The document discusses using yeast as a model system to study heterologous G protein-coupled receptors (GPCRs). Yeast have signaling pathways that can be exploited to study GPCRs. Chimeric Gα proteins were developed to efficiently couple yeast and mammalian GPCRs. Methods like modifying receptor sequences or co-expressing receptor activity modifying proteins allow functional expression of GPCRs in yeast. This enables large-scale screens that can define receptor-G protein specificity and identify novel ligands, improving drug discovery. Studies in yeast have characterized receptors, G proteins, and their regulators like receptor kinases and RGS proteins. Both Saccharomyces cerevisiae and Schizosaccharomyces pombe yeast are discussed
THEME – 4 Genomic diversity of domestication in soybeanICARDA
This document summarizes research on the genomic diversity of soybeans during domestication. It finds:
1) Wild soybeans (G. soja) show more genetic diversity than cultivated soybeans (G. max) due to bottlenecks during domestication and breeding.
2) Analysis of 7 G. soja genomes reveals a pan-genome structure with a large dispensable/variable gene set involved in environmental responses.
3) Comparison of G. soja and G. max genomes identified several million SNPs, indels, and presence/absence variants affecting genes related to domestication traits.
G Protein Coupled Receptor- An Introduction.Arindam Sain
The Largest family of integral membrane protein involved in many biological process and pathologies. 50% of all modern drugs and 25% of the top 200 best selling drugs are estimated to target GPCRs. Transduce the signals mediated by diverse signaling molecules, such as ions, peptides, lipids and photons, to induce different intracellular function. Bind their ligand and toa activate different G proteins.
Cloning and Expression of recombinant ProteinGaurav Dwivedi
Protein G was cloned from streptococcal strains and expressed in E. coli BL21 cells. Various expression and purification conditions were optimized, including inducer type, temperature, media, and heat treatment. Protein G was purified using immobilized metal affinity chromatography and its ability to bind IgG subclasses was confirmed using a VersaFLo system. Heat treatment at 80°C removed contaminants and was an efficient step in downstream processing. Overall, the study successfully cloned and expressed recombinant protein G and analyzed its IgG binding properties.
The researchers isolated and characterized the temperate mycobacteriophage Butters, which has one of the smallest known mycobacteriophage genomes at 41,491 bp. Using a technique called BRED, they deleted genes in the Butters genome, including the integrase gene gp37 and several orpham genes of unknown function. Deletion of the integrase gene resulted in mutant phages that formed larger, clearer plaques, suggesting a shift to a lytic life cycle. Deletion of the orpham genes gp30 and gp57 resulted in larger plaque sizes but did not prevent lysogeny, indicating these genes are not essential for the lysogenic cycle. Ongoing work includes further characterizing
Efectos potenciales a la salud por transgenicos y pesticidasCHST
1. The document discusses potential human health effects from genetically engineered crops and pesticides, including concerns about Bt crops and glyphosate.
2. Studies have found that Bt toxins from crops can survive digestion and negatively impact gut cells in mice. Bt toxins have also been shown to strongly stimulate the immune system.
3. Further testing is recommended to demonstrate the safety of Cry1A proteins and ensure genetically engineered crops do not pose risks to human health before they are commercialized.
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This document discusses molecular communication between parasitic plants and their hosts. It describes how parasitic plants identify hosts using strigolactones, locate hosts chemotropically, and form haustoria that penetrate the host and establish nutrient transfer. Parasitic plants and hosts exchange proteins and RNA, including microRNAs, to shape their interaction. The host mounts defense responses against haustoria like lignification and reactive oxygen species. Case studies examine how green peach aphid feeding on Cuscuta induces defenses in Cuscuta and systemic signals in the soybean host that increase its insect resistance.
This document discusses protein glycation in plants. It begins by defining protein glycation and the Maillard reaction, describing early and advanced glycation. It then discusses advanced glycation end products (AGEs) and mentions several types of AGEs that can form in plants, including levels found in different plant-derived foods. The document notes possible roles of protein glycation in plant physiology and stress responses. It focuses on methylglyoxal as a major glycating agent in plants and its role in plant processes. Finally, it discusses methylglyoxal detoxification pathways in plants involving glyoxalase enzymes and modifications to the glyoxalase system, as well as applications and future prospects regarding glycation in plants
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This document discusses plant disease outbreaks and epidemics. It defines an outbreak as a sudden increase in disease occurrences in a particular place and time, while an epidemic is a change in disease over time and space that can affect regions or countries. Epidemics develop from a combination of susceptible hosts, virulent pathogens, and favorable environmental conditions over a long period. Factors like host genetics, crop type, and moisture levels can influence epidemic development. The document also outlines management strategies like computer modeling, forecasting, and warning systems to help control epidemics.
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The cost of acquiring information by natural selectionCarl Bergstrom
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
The cost of acquiring information by natural selection
Heterotrimerric g protein can helpto feed the world
1. Heterotrimeric G protein: Can it help to feed
the world?
Upasana Mohapatra
PALB 6290
Dept. of Plant Biotechnology
UAS, GKVK, Bengaluru
2. CONTENTS
Summary and future prospectives
Case study
G protein signaling in plant stress responses
G protein signaling in plant physiological functions
G protein subunits
Structure and activation of G protein
History
April 13,2018 2Dept. of Plant Biotechnology,UAS(B)
3. HISTORY OF PLANT G-PROTEIN SCIENCE
Urano et al., 2013
April 13,2018 3Dept. of Plant Biotechnology,UAS(B)
4. Structure of G protein
• Heterotrimeric G protein signaling
regulates a wide range of growth
and developmental processes in
animals and plants.
• The membrane bound
heterotrimeric G protein complex
is composed of three subunits:
alpha (Gα), beta (Gβ), and
Gamma (Gγ) , G-protein-coupled
receptors (GPCRs), and regulators
of G-protein signaling proteins
(RGSs).
April 13,2018 4Dept. of Plant Biotechnology,UAS(B)
( Liu et al.,2017).
5. Activation of G protein
• Activation is mediated by
binding of an extracellular
ligand to a 7-pass trans-
membrane (7TM) G protein-
coupled receptor (GPCR).
• GPCRs are guanine-nucleotide
exchange factors; they cause Gα
to exchange GDP for GTP,
leading to the dissociation of the
complex into Gα -GTP and Gβγ
dimers.
April 13,2018 5Dept. of Plant Biotechnology,UAS(B)
( Liu et al.,2017).
6. The ‘G’ cycle of animals versus Arabidopsis
Urano et al., 2013
April 13,2018 6Dept. of Plant Biotechnology,UAS(B)
7. Crystal structure and activation mechanisms of G protein
Urano et al., 2013
April 13,2018 7Dept. of Plant Biotechnology,UAS(B)
8. Domain structures of Arabidopsis G protein-related proteins
Urano et al., 2013
April 13,2018 8Dept. of Plant Biotechnology,UAS(B)
9. Domain structures of Arabidopsis G protein-related proteins
Urano et al., 2013April 13,2018 9Dept. of Plant Biotechnology,UAS(B)
10. Structural characteristics of G protein signaling related proteins
Stateczny et al., 2016April 13,2018 10Dept. of Plant Biotechnology,UAS(B)
11. Structural characteristics of G protein signaling related proteins
Stateczny et al., 2016
April 13,2018 11Dept. of Plant Biotechnology,UAS(B)
12. Model for an intracellular kinase-dependent G
protein cycle in Arabidopsis
Stateczny et al., 2016
April 13,2018 12Dept. of Plant Biotechnology,UAS(B)
13. Hypothetic model for a membrane associated
kinase-dependent G protein mechanism in
Arabidopsis:
Stateczny et al., 2016
April 13,2018 13Dept. of Plant Biotechnology,UAS(B)
14. G protein subunits
In Plant cell
• In Arabidopsis-
one canonical Gα (AtGPA1)
one Gβ (AGβ1)
three Gγ(AGγ 1, AGγ 2,AGγ3)
• In rice-one canonical Gα , one
Gβ and 5 Gγ –subunits
• γ -subunits are four times the
average mammalian size.
• Many plant γ -subunits do not
contain an isoprenylation motif
at their C-terminus.
In Human beings
• 23 Gα, five Gβ, and 12 Gγ
subunits
• γ -subunits are very small
proteins (less than 100 amino
acids)
• An obligate requisite in all
animal γ -subunits and
essential for membrane
anchoring.
April 13,2018 14Dept. of Plant Biotechnology,UAS(B)
Stateczny et al., 2016
15. Physiological Functions of G Protein
Cell proliferation
Root development
Leaf and fruit development
Chloroplast development
Hormonal regulation of seed germination
Auxin and sugar signalling
ABA, Ca2+ and ROS signalling in guard cells.
Stomatal opening and closure
Phytochrome and cryptochrome mediated responses
April 13,2018 15Dept. of Plant Biotechnology,UAS(B) Stateczny et al., 2016
16. 1. Arabidopsis T-DNA insertion lines for Gα (gpa1–4), Gβ (agb1–2) or RGS1 (rgs1–2)
and wild-type Col-0 were grown for 37 days in a short day chamber (8 L : 16 D
cycle, 100 µmol m-2 s-1) at 230C.
2. Nipponbare (wild-type), the Gα knockout (d1,) or Gβ knockdown lines of rice were
grown in a short day chamber (8 L : 16 D cycle, 34 0C during day per 28 0C during
night time, 320 µmol m-2 s-1) for 47 days.
April 13,2018 16Dept. of Plant Biotechnology,UAS(B)
19. The α subunit encoded by a single gene (GPA1) in Arabidopsis, is a
modulator of plant cell proliferation.
1) gpa1 mutants have reduced cell division.
2) Inducible over-expression of GPA1 in Arabidopsis confers
inducible ectopic cell division which causes premature advance of the
nuclear cycle and the premature appearance of a division wall.
April 13,2018 19Dept. of Plant Biotechnology,UAS(B)
20. •Cell production rates were determined for the RAM and lateral root
formation in gpa1 and agb1 single and double mutants, and in transgenic
lines over expressing GPA1 or AGB1.
•In the RAM that the heterotrimeric complex acts as an attenuator of cell
proliferation, whereas Gα-subunit’s role is a positive modulator.
•For the formation of lateral roots, the Gβγ-dimer acts independently of
the Gα -subunit to attenuate cell division.
April 13,2018 20Dept. of Plant Biotechnology,UAS(B)
22. G Protein in Stress Response
Oxidative stress and hypoxia responses
Pathogen responses
Drought stress
Salinity stress
April 13,2018 22Dept. of Plant Biotechnology,UAS(B)
23. 1. TheAGB1 mutants (agb1-2-1 and agb1-3-2) were more sensitive to drought than
the wild-type.
2. The overexpression of mulberry (Morus alba L.) G-protein b subunit in transgenic
tobacco (Nicotiana tabacum L.) significantly enhanced the plants' drought
tolerance.
3. The transgenic tobacco plants had higher proline contents and peroxidase activities,
and lower malonaldehyde and hydrogen peroxide contents and superoxide free
radical accumulations under drought conditions and antioxidative genes, NtSOD
and NtCAT, are increased in drought-stressed transgenic tobacco plants
April 13,2018 23Dept. of Plant Biotechnology,UAS(B)
24. Overexpression of
mulberry (Morus alba
L.) G-protein β subunit
in transgenic tobacco
(Nicotiana tabacum L.)
TheAGB1 mutants (agb1-2-
1 and agb1-3-2) were more
sensitive
April 13,2018 24Dept. of Plant Biotechnology,UAS(B)
25. •The knockout mutant of the Arabidopsis heterotrimeric G-protein Gβ
subunit, agb1, is hypersensitive to salt, exhibiting a leaf bleaching
phenotype.
•AGB1 is mainly involved in the ionic toxicity component of salinity
stress and plays roles in multiple processes of Na+ homeostasis.
April 13,2018 25Dept. of Plant Biotechnology,UAS(B)
26. The knockout mutant
of the Arabidopsis
heterotrimeric G-
protein Gβ subunit,
agb1, is hypersensitive
to salt, exhibiting a leaf
bleaching phenotype.
Three-week-old
hydroponically grown
plants treated with 100
mm NaCl for 10 d.
Survival rate is less in
agb-1 mutant plants.
April 13,2018 26Dept. of Plant Biotechnology,UAS(B)
27. PAMP-triggered immunity
Cell death and defence responses
Signalling against bacterial pathogens
Elicitor-induced stomata closure
Resistance against necrotrophic fungi
Non-host resistance
R protein mediated disease resistance
Heterotrimeric G proteins in plant defence mechanisms
Nitta et al., 2015
April 13,2018 27Dept. of Plant Biotechnology,UAS(B)
Gβ
Gβ
Gα
Gβ,Gγ
Gα,Gβ
Gα,Gβ,Gγ
Gβ,Gγ
28. Defence-related phenotypes observed in heterotrimeric
G protein mutants
Nitta et al,2015
April 13,2018 28Dept. of Plant Biotechnology,UAS(B)
29. The Gα subunit is not involved in
the Gβ-mediated defense
pathway.
i) chorosis, and necrosis were
less developed in agb1-2
ii) inflorescence growth was also
inhibited in gpa1-4 plants,
although to a lesser extent than in
agb1-2; and
iii) endogenous SA did not affect
resistance to CMV and TuMV
and did not interfere with AGB1-
mediated responses.
April 13,2018 29Dept. of Plant Biotechnology,UAS(B)
31. Case study
1. The Gβ protein is essential for plant survival and growth.
2. Gα provides a foundation for grain size expansion.
3. Three Gγ proteins, DEP1, GGC2 and GS3, antagonistically regulate
grain size. DEP1 and GGC2, individually or in combination,
increase grain length when in complex with Gβ.
April 13,2018 31Dept. of Plant Biotechnology,UAS(B)
32. Grain size is a trait for grain quality focused by rice
breeders, as long and slender grains are preferred by rice
consumers in many countries
Recent advances in rice functional genomics facilitated
the cloning of a series of loci controlling grain size,
including genes
1) Grain length -GS3, GL3. An-1, GLW7, GS2 genes
2) Grain width -GW2, GW5, GS5, GW8 , GW7
3) Grain weight-GIF1, GE, TGW6, GW6a, BG1,XIAO
Introduction
April 13,2018 32Dept. of Plant Biotechnology,UAS(B)
33. Types of Gγ
Type A
• Small in size
• Has C-terminal
CaaX
isoprenylation
motif
• eg-RGG1-
regulation of
abiotic stress
Type B
• Small in size
• Lack CaaX motif
• eg-RGG2-
regulation of
abiotic stress
Type C
• 2 well defined
region-
• 1)an N-terminal
domain with
high similarity to
g subunit
• 2) A C terminal
cystein rich
subunit
• eg- DEP1,GS3-
QTL for grain
size.
April 13,2018 33Dept. of Plant Biotechnology,UAS(B)
34. Methods
1
• Field planting and trait measurement.
• Vector construction and plant transformation -Constructs of GS3-1OE,
GS3-4OE, DEP1OE, dep1OE and GGC2OE , under an ubiquitin gene
promoter.
2
• Generation of knock-out mutants
• Single mutant, double mutant and triple mutant of GS3,DEP1, GGC2
3
• RNA extraction and qRT-PCR.
• Western blot.
4
• Statistical analysis-The two-tailed t test was used for comparing agronomic
traits of each transgenic line with the control
April 13,2018 34Dept. of Plant Biotechnology,UAS(B)
35. •Elevated DEP1 accumulation
increased the grain length by
6.85–9.58% with a normal
plant stature.
•Plants over expressing dep1
showed very similar
phenotype to DEP1Ri plants,
both of which reduced grain
length by ~4.5%, together
with dwarf stature and erect
panicles.
•The expression level of endogenous
DEP1 in dep1OE was not reduced.
• Thus dep1 showed a dominant-negative
effect over DEP1 in regulating grain size
rather than co-suppression of the two
genes.April 13,2018 35Dept. of Plant Biotechnology,UAS(B)
36. •GS3-1OE showed an
average 9.07% reduction in
grain length together with
reduced plant stature.
•GS3-4OE produced even
smaller plants and grain size
with an average 19.10%
reduction of grain length
.
•GS3-1Ri resulted in an
average 5.78% increase of
grain length.
April 13,2018 36Dept. of Plant Biotechnology,UAS(B)
37. April 13,2018 Dept. of Plant Biotechnology,UAS(B) 37
GS3-1Ri X dep1OE
F1 Reduced grain length, similar to
dep1OE transgenic plants.
F2
Also showed the dep1OE phenotype with
respect to grain size, plant height and
panicle length
Interaction between DEP1 and GS3 in grain size regulation
38. GS3-1ko mutant-increased grain size,
DEP1ko mutant - reduced grain size.
GS3-1koDEP1ko double mutant- intermediate
between that of the single mutants of GS3-1ko
and DEP1ko
GS3-1koDEP1ko double mutant X ZH11
F2
Increased grain length was observed when
over expressing DEP1 in GS3-1Ri
background,
whereas the DEP1OE/GS3-1OE hybrid
showed the GS3-1OE phenotype of short
grain
April 13,2018 38Dept. of Plant Biotechnology,UAS(B)
39. An atypical Gγ subunit GGC2 functions additively with DEP1
Over expression of GGC2 (GGC2OE) in ZH11
increased grain length significantly,
knock-out mutant of GGC2ko in ZH11 reduced the
grain length.
Knocking out DEP1ko GGC2ko resulted in much smaller
grains than either of the single knock-out mutants,
DEP1 and GGC2 worked additively in positive regulation of
grain length.
The DEP1koGGC2koGS3-1ko triple mutant
produced similar phenotype to that of
DEP1koGGC2ko, thus GS3-1ko mutant could not
increase grain length when both DEP1 and
GGC2 were knocked-out
DEP1 and GGC2 functioned positively in regulating grain size in an additive
manner, while the role of GS3 in grain size regulation was to repress the effects
of DEP1 and GGC2 on increasing grain size.
April 13,2018 39Dept. of Plant Biotechnology,UAS(B)
40. Functional dependence of the Gγ subunits on
RGB1 and RGA1.
The transgenic plants with suppressed expression of RGB1 (Knocked down RGB1 by RNAi
(RGB1Ri) )showed reduced grain size and plant height.
GS3-1Ri X RGB1Ri DEP1OE X RGB1Ri
Reduced grain length
Reduced grain length
Overexpressed GGC2 in RGB1Ri.
Overexpressed DEP1 in RGB1Ri.
Reduced grain length
The effects of grain
length increase by
DEP1OE, GGC2OE and
GS3-1Ri are dependent
on RGB1 and RGA1
April 13,2018 40Dept. of Plant Biotechnology,UAS(B)
41. Schematic representation of the functions of the G proteins in grain size regulation
April 13,2018 41Dept. of Plant Biotechnology,UAS(B)
42. Conclusion
• DEP1 and GGC2, when coupled with RGB1, promote
grain size by tail-mediated signaling.
• GS3,reduces grain size by blocking the interaction of
DEP1 and GGC2 with RGB1.
• The increased grain size is at the cost of fitness. Thus
from fitness viewpoint medium grain is the optimal for
rice reproduction.
• Thus, while DEP1 and GGC2 function to promote grain
size,which is essential for yield increase in breeding,
GS3 plays a role to keep balance for maintaining fitness
in response to natural selection.
April 13,2018 42Dept. of Plant Biotechnology,UAS(B)
43. 1) The interactions of three Gγ proteins can be used for a predictable design of
grain size in rice, by manipulating these genes, individually or in combination, to
improve rice grain yield and quality.
2) As the Gγ proteins are highly conserved in a very wide range of plants,
manipulating these proteins may provide a general strategy for modifying organ
size and yield in crop breeding.
3) G signalling is at the heart of many plant physiologies of agronomic
importance, such as disease resistance, abiotic stress tolerance and harvest index.
Translational work on G protein signalling will certainly improve agriculture by
providing new targets and strategies for increasing yield.
April 13,2018 43Dept. of Plant Biotechnology,UAS(B)
44. Future Prospectives
• What are the receptors upstream of G-proteins, what lies
downstream of G-proteins, and how the proteins connect to the
established modules of hormone, defense or stress-related
signaling.
• Although the G proteins have been shown to have key roles in
regulating plant growth, stress tolerance and grain yield
potential, the molecular mechanisms underlying the
regulation of the heterotrimeric G protein cycles of activation
and deactivation and downstream effectors still remain
unclear.
April 13,2018 44Dept. of Plant Biotechnology,UAS(B)
45. April 13,2018 Dept. of Plant Biotechnology,UAS(B) 45
THANK YOU
Science of G protein is awaiting your
research efforts, to be explored…….
46. REFERENCES
BOTELLA, J.R., 2012, Can heterotrimeric G proteins help to feed the world?. Trends in
plant science, 17(10) : 563-568.
CHEN, J.G., GAO, Y. AND JONES, A.M., 2006, Differential roles of Arabidopsis
heterotrimeric G-protein subunits in modulating cell division in
roots. Plant Physiology, 141(3), pp.887-897.
LIU, C., XU, Y., LONG, D., CAO, B., HOU, J., XIANG, Z. AND ZHAO, A., 2017,
Plant G-protein β subunits positively regulate drought tolerance by
elevating detoxification of ROS. Biochemical and biophysical research
communications, 491(4): 897-902.
STATECZNY, D., OPPENHEIMER, J. AND BOMMERT, P., 2016, G protein signaling
in plants: minus times minus equals plus .Current opinion in plant
biology, 34 :127-135.
SUN, S., WANG, L., MAO, H., SHAO, L., LI, X., XIAO, J., OUYANG, Y. AND
ZHANG, Q., 2018, A G-protein pathway determines grain size in
rice. Nature communications, 9(1): 851.
URANO, D., CHEN, J.G., BOTELLA, J.R. AND JONES, A.M., 2013, Heterotrimeric
G protein signalling in the plant kingdom. Open biology, 3(3): 120186.
XU, Q., ZHAO, M., WU, K., FU, X. AND LIU, Q., 2016, Emerging insights into
heterotrimeric G protein signaling in plants. Journal of Genetics and
Genomics, 43(8): 495-502.
April 13,2018 46Dept. of Plant Biotechnology,UAS(B)
Editor's Notes
History of plant G protein science. In the 1970s, G proteins were identified as a signal transducer connecting the hormone receptor and the adenylyl cyclase in mammals. In the early 1990s, plant G protein genes were cloned and shown to have conserved domains and motifs with the animal genes. In the late 1990s, much effort went towards physiological roles of G proteins using genetics. In the 2000s, the Gβγ-subunits, the regulators (GPCR-like genes and a 7TM-RGS gene) and effectors of G protein were cloned and characterized genetically and biochemically. In 2007, the ‘self-activating’ property of the plant G protein was revealed. In addition, the transcriptome, proteome and interactome analyses revealed comprehensive knowledge of the plant G protein pathways. In the last few years, the crystal structure and computational simulation solved the mechanism of self-activation. Publications on the physiological functions and signalling components of G protein pathways are exponentially increasing, providing evidence for their important and divergent functions in plants.
These two functional subunits activate different signaling cascades until the intrinsic GTPase activity of the Gα subun
RGS-Regulator of G proteins signaling
The ‘G’ cycle of animals versus Arabidopsis. (a) G protein regulation in mammalian cells. In the absence of liGα nd, G protein forms an inactive heterotrimer with Gβγ dimer (bottom left). LiGα nd-bound GPCR promotes GDP dissociation and GTP binding on G protein (top). GTP-bound Gα dissociates from Gβγ dimer, and both activated Gα and freely released Gβγ modulate activity of the effectors (bottom right). Gα hydrolyses GTP to GDP, and re-binds to Gβγ to return to its inactive state. (b) G protein regulation modelled in Arabidopsis. Arabidopsis G protein (AtGPA1) can spontaneously dissociate GDP and activate itself (bottom left). AtGPA1 does not hydrolyse its GDP rapidly; however, AtRGS1, a 7TM-RGS protein, promotes the GTP hydrolysis of AtGPA1 (top). d-glucose or other stimuli functions on AtRGS1 directly or indirectly, and decouples AtGPA1 from AtRGS1 (bottom right). Once released from AtRGS1, AtGPA1 does not hydrolyse its GTP efficiently, maintaining its active state and modulating the effector activities.
Crystal structure and activation mechanisms of G protein. (a) Structural basis of animal G protein activation. Left: Gα protein forms stable heterotrimer
with Gβg dimer (grey and black) at the steady state. GDP (green) is tightly bound to a Ras domain (orange) of the a-subunit, and covered by the helical domain
(sky blue). Right: in the presence of liGα nd-bound receptor, the helical domain moves and changes orientation. The structural change causes GDP dissociation from
the a-subunit, the subsequent GTP binding and activation. (b) Structure of Arabidopsis AtGPA1 is entirely similar to mammalian Gα proteins. However, the helical
domain of AtGPA1 fluctuates spontaneously. The spontaneous fluctuation initiates GDP dissociation, and nucleotide exchange. Crystal structures shown are animal
heterotrimeric G protein (PDB: 1GOT) [56], G protein and b2 adrenergic receptor (PDB: 3SN6) [6], and Arabidopsis AtGPA1 (PDB: 2XTZ) [41]. The cartoon for the
animal model was adapted from Rasmussen et al. [6].
1) AtRGS1 keeps the heterotrimeric complex in its inactive state by preventing the exchange of GDP to GTP.
(2) D-Glucose induction promotes the phosphorylation of AtRGS1 by WNK8 (intracellular kinase)
(3) Phosphorylated AtRGS1 undergoes endocytosis and the uncoupled Gα protein is capable to exchange GDP for GTP. GTP bound Gα and the detached Gβγ dimer can activate downstream effectors.
(1) AtRGS1 keeps the heterotrimeric complex in its inactive state by preventing the exchange of GDP to GTP.
(2) Upon binding of a liGα nd, the RLK dimer phosphorylates AtRGS1.
(3) Phosphorylated AtRGS1 undergoes endocytosis and the uncoupled Gα protein is capable to exchange GDP for GTP. GTP bound Gα and the detached Gβγ dimer can activate downstream effectors.
G protein g-subunits exhibit an extraordinary level of
structural diversity
These results suggest that
Arabidopsis heterotrimeric G-protein subunits have differential and opposing roles in the modulation of cell division in roots
These results suggest that
Arabidopsis heterotrimeric G-protein subunits have differential and opposing roles in the modulation of cell division in roots
Cell death is regulated by AGB1 ..as agb1 is not there there will be stoppage of cell growth and no necrosis and chlorosis.so the defence is regulated by Gb.
SA is not involved inhis regulation pathway