Plant Genetics, NGS and Bioinformatics
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50 Top Sources Of Free eLearning Courses

50 Top Sources Of Free eLearning Courses | Plant Genetics, NGS and Bioinformatics | Scoop.it

Whether you are looking for a master’s degree program, computer science classes, a K-12 curriculum, or GED study program, this list gives you50 Top Sources Of Free eLearning Courses - 


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Plant Genetics, NGS and Bioinformatics
Papers and topics in plant genetics, NGS and bioinformatics
Curated by Ali Taheri
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Rescooped by Ali Taheri from Plant and Seed Biology
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TILLING by Sequencing (TbyS) for targeted genome mutagenesis in crops

TILLING by Sequencing (TbyS) for targeted genome mutagenesis in crops | Plant Genetics, NGS and Bioinformatics | Scoop.it
TILLING (Targeting Induced Local Lesions in Genomes) by Sequencing (TbyS) refers to the application of high-throughput sequencing technologies to mutagenised TILLING populations as a tool for function

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Rescooped by Ali Taheri from Publications from The Sainsbury Laboratory
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Ann Rev Phytopath: Function, Discovery, and Exploitation of Plant Pattern Recognition Receptors for Broad-Spectrum Disease Resistance (2017)

Ann Rev Phytopath: Function, Discovery, and Exploitation of Plant Pattern Recognition Receptors for Broad-Spectrum Disease Resistance (2017) | Plant Genetics, NGS and Bioinformatics | Scoop.it

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The Sainsbury Lab's curator insight, June 19, 7:04 AM
Plants are constantly exposed to would-be pathogens and pests, and thus have a sophisticated immune system to ward off these threats, which otherwise can have devastating ecological and economic consequences on ecosystems and agriculture. Plants employ receptor kinases (RKs) and receptor-like proteins (RLPs) as pattern recognition receptors (PRRs) to monitor their apoplastic environment and detect non-self and damaged-self patterns as signs of potential danger. Plant PRRs contribute to both basal and non-host resistances, and treatment with pathogen-/microbe-associated molecular patterns (PAMPs/MAMPs) or damage-associated molecular patterns (DAMPs) recognized by plant PRRs induces both local and systemic immunity. Here, we comprehensively review known PAMPs/DAMPs recognized by plants as well as the plant PRRs described to date. In particular, we describe the different methods that can be used to identify PAMPs/DAMPs and PRRs. Finally, we emphasize the emerging biotechnological potential use of PRRs to improve broad-spectrum, and potentially durable, disease resistance in crops.
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Getting the Hologenome Concept Right: an Eco-Evolutionary Framework for Hosts and Their Microbiomes

Getting the Hologenome Concept Right: an Eco-Evolutionary Framework for Hosts and Their Microbiomes | Plant Genetics, NGS and Bioinformatics | Scoop.it
Given the complexity of host-microbiota symbioses, scientists and philosophers are asking questions at new biological levels of hierarchical organization—what is a holobiont and hologenome? When should this vocabulary be applied? Are these concepts a null hypothesis for host-microbe systems or limited to a certain spectrum of symbiotic interactions such as host-microbial coevolution? Critical discourse is necessary in this nascent area, but productive discourse requires that skeptics and proponents use the same lexicon. For instance, critiquing the hologenome concept is not synonymous with critiquing coevolution, and arguing that an entity is not a primary unit of selection dismisses the fact that the hologenome concept has always embraced multilevel selection. Holobionts and hologenomes are incontrovertible, multipartite entities that result from ecological, evolutionary, and genetic processes at various levels. They are not restricted to one special process but constitute a wider vocabulary and framework for host biology in light of the microbiome.

Via Francis Martin, Loïc Lepiniec
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Nature Biotech: A pigeonpea gene confers resistance to Asian soybean rust in soybean (2016)

Nature Biotech: A pigeonpea gene confers resistance to Asian soybean rust in soybean (2016) | Plant Genetics, NGS and Bioinformatics | Scoop.it

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.


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The Sainsbury Lab's curator insight, April 26, 2016 4:45 AM
Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.
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PITCh: An Alternative Use of TALENs and CRISPR/Cas9

PITCh: An Alternative Use of TALENs and CRISPR/Cas9 | Plant Genetics, NGS and Bioinformatics | Scoop.it
A new gene insertion method called Precise Integration into Target Chromosome (PITCh) uses existent gene editing technologies more effectively thanks to MMEJ.
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Microbiomes: Curating communities from plants : Nature : Nature Publishing Group

Microbiomes: Curating communities from plants : Nature : Nature Publishing Group | Plant Genetics, NGS and Bioinformatics | Scoop.it

Large-scale cultivation and genome sequencing of the bacteria that inhabit the leaves and roots of Arabidopsis plants have paved the way for probing how microbial communities assemble and function. 


Via Niklaus Grunwald, Guogen Yang
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Biologists create more precise molecular scissors for genome editing

Biologists create more precise molecular scissors for genome editing | Plant Genetics, NGS and Bioinformatics | Scoop.it
Engineered Cas9 enzyme makes fewer mistakes.

Via Loïc Lepiniec
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DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins

DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins | Plant Genetics, NGS and Bioinformatics | Scoop.it
Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

Via Biswapriya Biswavas Misra, Loïc Lepiniec
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Biswapriya Biswavas Misra's curator insight, November 5, 2015 9:45 PM

Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

Rescooped by Ali Taheri from Ag Biotech News
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Next biotech plants: new traits, crops, developers and technologies for addressing global challenges - Ricroch & Henard (2015) - GMCC

Most of the genetically modified (GM) plants currently commercialized encompass a handful of crop species (soybean, corn, cotton and canola) with agronomic characters (traits) directed against some biotic stresses (pest resistance, herbicide tolerance or both) and created by multinational companies.

 

The same crops with agronomic traits already on the market today will continue to be commercialized, but there will be also a wider range of species with combined traits. The timeframe anticipated for market release of the next biotech plants will not only depend on science progress... but also primarily on how demanding regulatory requirements are in countries where marketing approvals are pending.

 

Regulatory constraints, including environmental and health impact assessments, have increased significantly in the past decades, delaying approvals and increasing their costs. This has sometimes discouraged public research entities and small and medium size plant breeding companies from using biotechnology and given preference to other technologies, not as stringently regulated.

 

Nevertheless R&D programs are flourishing in developing countries, boosted by the necessity to meet the global challenges that are food security of a booming world population while mitigating climate change impacts. Biotechnology is an instrument at the service of these imperatives and a wide variety of plants are currently tested for their high yield despite biotic and abiotic stresses...

 

Food security is not only a question of quantity but also of quality of agricultural and food products, to be available and accessible for the ones who need it the most. Many biotech plants (especially staple food) are therefore being developed with nutritional traits, such as biofortification in vitamins and metals.

 

The main international seed companies continue to be the largest investors in plant biotechnology R&D, and often collaborate in the developing world with public institutions, private entities and philanthropic organizations. These partnerships are particularly present in Africa. 

 

In developed countries, plant biotechnology is also used for non-food purposes, such as the pharmaceutical, biofuel, starch, paper and textile industries...

 

Various plant breeding technologies are now used in the entire spectrum of plant biotechnology... Next generation precision gene editing tools are developed in basic research. They include: clustered regularly interspaced short palindromic repeats (CRISPR), oligonucleotide- directed mutagenesis (ODM), transcription activator-like effects nucleases (TALENs) and zinc-finger nuclease (ZFN).

 

http://purl.umn.edu/211484


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Mass spectrometry imaging for plant biology: a review

Mass spectrometry imaging for plant biology: a review | Plant Genetics, NGS and Bioinformatics | Scoop.it
Mass spectrometry imaging (MSI) is a developing technique to measure the spatio-temporal distribution of many biomolecules in tissues. Over the preceding decade, MSI has been adopted by plant biologists and applied in a broad range of areas, including primary metabolism, natural products, plant defense, plant responses to abiotic and biotic stress, plant lipids and the developing field of spatial metabolomics. This review covers recent advances in plant-based MSI, general aspects of instrumentation, analytical approaches, sample preparation and the current trends in respective plant research.

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Eric Vincill's curator insight, October 18, 2015 4:09 PM

The detection of biomolecules in plant tissues  by mass spec. LOVE IT!! I can't tell you how excited I am to see protocols being established that increase the spatiotemporal resolution of the movement of small molecules in the plant.  

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An interdisciplinary shift in demand for talent within the biotech industry : Nature Biotechnology : Nature Publishing Group

An interdisciplinary shift in demand for talent within the biotech industry : Nature Biotechnology : Nature Publishing Group | Plant Genetics, NGS and Bioinformatics | Scoop.it
A shift away from hiring narrowly focused specialists to individuals with interdisciplinary academic training highlights the latest life sciences workforce trends.
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Rescooped by Ali Taheri from The Plant Microbiome
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Frontiers: Primer and platform effects on 16S rRNA tag sequencing

Frontiers: Primer and platform effects on 16S rRNA tag sequencing | Plant Genetics, NGS and Bioinformatics | Scoop.it

Sequencing of 16S rRNA gene tags is a popular method for profiling and comparing microbial communities. The protocols and methods used, however, vary considerably with regard to amplification primers, sequencing primers, sequencing technologies; as well as quality filtering and clustering. How results are affected by these choices, and whether data produced with different protocols can be meaningfully compared, is often unknown. Here we compare results obtained using three different amplification primer sets (targeting V4, V6–V8, and V7–V8) and two sequencing technologies (454 pyrosequencing and Illumina MiSeq) using DNA from a mock community containing a known number of species as well as complex environmental samples whose PCR-independent profiles were estimated using shotgun sequencing. We find that paired-end MiSeq reads produce higher quality data and enabled the use of more aggressive quality control parameters over 454, resulting in a higher retention rate of high quality reads for downstream data analysis. While primer choice considerably influences quantitative abundance estimations, sequencing platform has relatively minor effects when matched primers are used. Beta diversity metrics are surprisingly robust to both primer and sequencing platform biases.


Via Stéphane Hacquard
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Rescooped by Ali Taheri from Plant and Seed Biology
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Chinese Scientists Created Transgenic Crops that Resist Heat While Increase Yields -

Chinese Scientists Created Transgenic Crops that Resist Heat While Increase Yields - | Plant Genetics, NGS and Bioinformatics | Scoop.it
An essay published on Nature Biotechnology on August 17 reveals an exciting achievement of Chinese scientists: they have successfully cultivated transgenic crops with an ERECTA gene (henceforth ER) that can resist heat while increase production.

Researchers from Institute of Plant Physiology & Ecology, SIBS, CAS, together contribute to this essay entitled “Overexpression of Receptor-like Kinase ERECTA Improves Thermotolerance in Rice and Tomato”.

Via Christophe Jacquet, Loïc Lepiniec
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Rescooped by Ali Taheri from Plant roots and rhizosphere
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Understanding and exploiting plant beneficial microbes

Understanding and exploiting plant beneficial microbes | Plant Genetics, NGS and Bioinformatics | Scoop.it

Highlights

• Soil microbiomes induce reproducible plant phenotypes.
• Large collections of plant-associated microbes are available for research.
• Plant growth promoting microbial inoculants can persist in soil for weeks.
• Co-regulation of immune system function and nutritional stress responses exists.
•Deployment of consortia may enable more resilient plant phenotypes than single strains.

After a century of incremental research, technological advances, coupled with a need for sustainable crop yield increases, have reinvigorated the study of beneficial plant–microbe interactions with attention focused on how microbiomes alter plant phenotypes. We review recent advances in plant microbiome research, and describe potential applications for increasing crop productivity. The phylogenetic diversity of plant microbiomes is increasingly well characterized, and their functional diversity is becoming more accessible. Large culture collections are available for controlled experimentation, with more to come. Genetic resources are being brought to bear on questions of microbiome function. We expect that microbial amendments of varying complexities will expose rules governing beneficial plant–microbe interactions contributing to plant growth promotion and disease resistance, enabling more sustainable agriculture.


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Online Statistics Education: A Free Resource for Introductory Statistics

Online Statistics Education: A Free Resource for Introductory Statistics | Plant Genetics, NGS and Bioinformatics | Scoop.it

Online Statistics: An Interactive Multimedia Course of Study is a resource for learning and teaching introductory statistics. It contains material presented in textbook format and as video presentations. This resource features interactive demonstrations and simulations, case studies, and an analysis lab.


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Nature Biotech: Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture (2016)

Nature Biotech: Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture (2016) | Plant Genetics, NGS and Bioinformatics | Scoop.it

Wild relatives of domesticated crop species harbor multiple, diverse, disease resistance (R) genes that could be used to engineer sustainable disease control. However, breeding R genes into crop lines often requires long breeding timelines of 5–15 years to break linkage between R genes and deleterious alleles (linkage drag). Further, when R genes are bred one at a time into crop lines, the protection that they confer is often overcome within a few seasons by pathogen evolution1. If several cloned R genes were available, it would be possible to pyramid R genes2 in a crop, which might provide more durable resistance1. We describe a three-step method (MutRenSeq)-that combines chemical mutagenesis with exome capture and sequencing for rapid R gene cloning. We applied MutRenSeq to clone stem rust resistance genes Sr22 and Sr45 from hexaploid bread wheat. MutRenSeq can be applied to other commercially relevant crops and their relatives, including, for example, pea, bean, barley, oat, rye, rice and maize.


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The Sainsbury Lab's curator insight, April 26, 2016 4:14 AM
Wild relatives of domesticated crop species harbor multiple, diverse, disease resistance (R) genes that could be used to engineer sustainable disease control. However, breeding R genes into crop lines often requires long breeding timelines of 5–15 years to break linkage between R genes and deleterious alleles (linkage drag). Further, when R genes are bred one at a time into crop lines, the protection that they confer is often overcome within a few seasons by pathogen evolution1. If several cloned R genes were available, it would be possible to pyramid R genes2 in a crop, which might provide more durable resistance1. We describe a three-step method (MutRenSeq)-that combines chemical mutagenesis with exome capture and sequencing for rapid R gene cloning. We applied MutRenSeq to clone stem rust resistance genes Sr22 and Sr45 from hexaploid bread wheat. MutRenSeq can be applied to other commercially relevant crops and their relatives, including, for example, pea, bean, barley, oat, rye, rice and maize.
Neelam Redekar's curator insight, April 29, 2016 8:19 AM
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Rescooped by Ali Taheri from Viruses and Bioinformatics from Virology.uvic.ca
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Scientists remove HIV-1 from genome of human immune cells

Scientists remove HIV-1 from genome of human immune cells | Plant Genetics, NGS and Bioinformatics | Scoop.it
Scientists edited HIV-1 DNA out of the genome of human immune cells, preventing virus replication and reinfection of the cleared cells.

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Rescooped by Ali Taheri from Viruses and Bioinformatics from Virology.uvic.ca
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How to build a better PhD

How to build a better PhD | Plant Genetics, NGS and Bioinformatics | Scoop.it
There are too many PhD students for too few academic jobs — but with imagination, the problem could be solved.

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The Gene Editing Tsunami | PLOS Synthetic Biology Community

The Gene Editing Tsunami | PLOS Synthetic Biology Community | Plant Genetics, NGS and Bioinformatics | Scoop.it
The Gene Editing Tsunami

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Swedish Board of Agriculture: a CRISPR-Cas9 mutant is not a GMO - Umeå Univ (2015)

Swedish Board of Agriculture: a CRISPR-Cas9 mutant is not a GMO - Umeå Univ (2015) | Plant Genetics, NGS and Bioinformatics | Scoop.it

The Swedish Board of Agriculture has... confirmed the interpretation that some plants in which the genome has been edited using the CRISPR-Cas9 technology do not fall under the European GMO definition. This is important for the wide use of such plants to contribute to solving some of the escalating challenges of mankind.

CRISPR-Cas9 is a technique... allowing scientists to make small edits in the genetic material of an organism, edits that can also occur naturally. Instead of hoping that such edits occur by natural recombination, they can now be deliberately introduced in a targeted and precise manner. CRISPR-Cas9 can thus be used in many ways in plant science and breeding.

 

Plants that fall within the scope of EU GMO legislation are subject to a very strict regulatory regime (in reality making it impossible to grow them in the field in most EU countries). Plants that fall outside the scope can be grown without restriction. Since “inside or outside of the GMO definition” will decide whether or not plant scientists will be able to use the technique for practical applications, plant scientists and breeders have been waiting for the authorities’ decision concerning CRISPR-Cas9... 

 

Countries such as Argentina have announced that similarly edited plants fall outside their GMO legislation, but no decision has been taken yet inside the EU. A complicating factor is that the technique can be used in several different ways with the consequence that some of the resulting plants may fall outside while others may fall inside the GMO legislation. Now, for the first time, concrete examples have been evaluated by a competent authority, and the Swedish Board of Agriculture announced... that some Arabidopsis plants that have been modified using CRISPR-Cas9 fall within the scope of the legislation while others do not... 

 

“What we now have done pinpoints the problem; using CRISPR-Cas9 we can create a plant that in ALL aspects is identical to one that is not considered to be a GMO. Common sense and scientific logic says that it is impossible to have two identical plants where growth of one is, in reality, forbidden while the other can be grown with no restrictions; how would a court be able to decide if the cultivation was a crime or not? But regulatory logic is not necessarily the same as scientific logic, and it is therefore important that the Swedish Board of Agriculture has interpreted the definition in this way”...  

 

This interpretation opens up the possibility that this technique can be used to address some of the biggest challenges for mankind, expressed in the sustainable development goals recently suggested by the United Nations... “We hope that this clear and logical interpretation will also be applied to other similar cases.

 

The EU commission announced some time ago that it would present its interpretation of the legislation, but has not yet been able to come to an agreement. All ‘GMO issues’ divide the EU and this has led to paralysis for more than a decade. We think that the opinion by the Swedish Board of Agriculture will get a lot of international attention”...  

 

http://www.umu.se/english/about-umu/news-events/news/newsdetailpage/green-light-in-the-tunnel-opinion-of-the-swedish-board-of-agriculture--a-crispr-cas9-mutant-but-not-a-gmo.cid259265

 


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Rescooped by Ali Taheri from Plant Biology Teaching Resources (Higher Education)
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Promoting microbiology education through the iGEM synthetic biology competition

Promoting microbiology education through the iGEM synthetic biology competition | Plant Genetics, NGS and Bioinformatics | Scoop.it

"Promoting microbiology education through the iGEM synthetic biology competition." I'm a huge fan of iGEM and this article summarizes all that it has achieved (so far). It's not just promoting microbiology, it's promoting enthusiasm in science more broadly and a really effective way to show undergraduates (and even high school students) what research is like.
http://femsle.oxfordjournals.org/conte…/…/16/fnv129.abstract


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Rescooped by Ali Taheri from Plant Biology Teaching Resources (Higher Education)
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Teenagers and mutant tomatoes

Teenagers and mutant tomatoes | Plant Genetics, NGS and Bioinformatics | Scoop.it

Wake Forest biology professors Gloria Muday and Carole Gibson and their students use mutant tomatoes grown in the campus garden to teach fundamental biology concepts to local high school students. "Teaching with Tomatoes" is funded by the American Society of Plant Biology Education Foundation and has evolved over the years.


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CRISPR tweak may help gene-edited crops bypass biosafety regulation

CRISPR tweak may help gene-edited crops bypass biosafety regulation | Plant Genetics, NGS and Bioinformatics | Scoop.it

A twist on a revolutionary gene-editing technique may make it possible to modify plant genomes while sidestepping national biosafety regulations, South Korean researchers say.


Plant scientists have been quick to experiment with the popular CRISPR/Cas9 technique, which uses an enzyme called Cas9, guided by two RNA strands, to precisely cut segments of DNA in a genome. By disabling specific genes in wheat and rice, for example, researchers hope to make disease-resistant strains of the crops.


But the process can introduce bits of foreign DNA into plant genomes. And some jurisdictions, such as the European Union, could decide to classify such plants as genetically modified organisms (GMOs)1 — making their acceptance by regulatory bodies contentious, says geneticist Jin-Soo Kim of Seoul National University.


Kim and his team tweaked the technique so that it can delete specific plant genes without introducing foreign DNA, creating plants that he and his colleagues think “might be exempt from current GMO regulations”2.


“In terms of science, our approach is just another improvement in the field of genome editing. However, in terms of regulations and public acceptance, our method could be path-breaking,” says Kim.


Conventionally, researchers get CRISPR/Cas9 working in a plant cell by first shuttling in the gene that codes for the Cas9 enzyme. The gene is introduced on a plasmid — a circular packet of DNA — which is usually carried into a plant by the bacterial pest Agrobacterium tumefaciens. As a result, Agrobacterium DNA can end up in the plant’s genome. Even if the pest is not used, fragments of the Cas9 gene may themselves be incorporated into the plant's genome.


To get around this problem, Kim and his colleagues avoid gene-shuttling altogether. They report a recipe to assemble the Cas9 enzyme together with its guide RNA sequences (which the enzyme requires to find its target) outside the plant, and use solvents to get the resulting protein complex into the plant. The technique works efficiently to knock out selected genes in tobacco plants, rice, lettuce and thale cress, they say, reporting their results in Nature Biotechnology2.


“I think this is a milestone work for plant science,’ says bioethicist Tetsuya Ishii at Hokkaido University in Sapporo, Japan, who has extensively studied the framework of regulation surrounding genetic engineering in plants.


Kim wants to use the technique to edit the banana; the crop's most popular cultivar, the Cavendish variety, is struggling to combat a devastating soil fungus and may go extinct. Gene editing could, for example, be used to knock out the receptor that the fungus uses to invade cells, without any need, in Kim’s view, to classify the resulting banana as a GMO. “We will save the banana so that our children and grandchildren can still enjoy the fruit,” he says.


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9 Facts About Biodiesel That Will Rev You Up - YouTube

Biodiesel works as a clean-burning, renewable alternative to petroleum diesel, offering a viable opportunity to diversify the U.S. transportation fuel supply...
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How will we fill 9 billion bowls by 2050? | #9billionbowls | Thomson Reuters

How will we fill 9 billion bowls by 2050? | #9billionbowls | Thomson Reuters | Plant Genetics, NGS and Bioinformatics | Scoop.it
Filling 9 Billion Bowls from Thomson Reuters tells the story of a diverse group of scientists, students, analysts and inventors who are using Big Data and leading edge technologies to face the challenge of feeding 9 billion people in 2050, and revolutionise the way we produce and deliver food.
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