CROP BIOTECH UPDATE
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A weekly summary of world developments in agri-biotech for developing countries, produced by the Global Knowledge Center on Crop Biotechnology, International Service for the Acquisition of Agri-biotech Applications SEAsiaCenter (ISAAA)
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November 7, 2018

In This Week’s Issue:

News

Global
• 13 WTO Members Support Policy Approaches to Enable Innovation in Agriculture

Africa
• Farmers and University Students Call for Urgent Action on Biotech Legislation in Uganda

Americas
• US Patent Awarded to DNA-Targeting Complex
• High Temperatures Can Trigger a Reaction in a Plant's RNA
• ASTA Lauds International Statement Supporting of Plant Breeding Innovation

Asia and the Pacific
• Scientists Discover Gene Regulator that Allows Plant to Rehydrate After Drought
• FAS Jakarta Launches "Biotech Ambassadors" Outreach
• USDA: The Philippines Remains as Asia's Leader in Biotech

Europe
• Gene Discovery Could Pave Way for Disease Resistant Crops
• Small Genetic Differences Make Plants into Better Teams
• Agriculture Minister of The Netherlands Opens Door to Genetic Modification

Research
• Protoplast Isolation Method for Genetic Improvement of Pineapple
• Gene from Castor Bean Increases Unusual Oil in Camelina

New Breeding Technologies
• Researchers Use CRISPR for Apple and Grapevine Improvement
• Plasmid-Free Genome Editing of Cabbage and Chinese Cabbage
• Plant Genome Editing Database (PGED) Goes Live

Beyond Crop Biotech
• Scientists Reveal a Gene Related to Alzheimer's Disease Using CRISPR

Document Reminders
• Pocket K: Contributions of Agri-biotech in Alleviation of Poverty and Hunger (Updated)



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NEWS
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Global
13 WTO MEMBERS SUPPORT POLICY APPROACHES TO ENABLE INNOVATION IN AGRICULTURE

The International Statement on Agricultural Applications of Precision Biotechnology was released in Geneva by the World Trade Organization (WTO) Committee on Sanitary and Phytosanitary Measures on October 26, 2018. At the Committee's meeting on November 1-2, 2018, WTO members discussed the role that precision biotechnology techniques can play in agricultural innovation, with a view to providing farmers around the world with access to tools that increase productivity while preserving environmental sustainability.

The communication is being circulated at the request of delegations from Argentina, Australia, Brazil, Canada, the Dominican Republic, Guatemala, Honduras, Paraguay, the United States of America, and Uruguay. Thirteen member countries (Argentina, Australia, Brazil, Canada, Colombia, The Dominican Republic, Guatemala, Honduras, Jordan, Paraguay, Uruguay, the United States of America and Uruguay), 10 of each planted biotech crops in 2017, have supported the international statement. The effort in drafting the statement began in Argentina during the "Seminar on Genome Editing for Regulators", organized by the Inter-American Institute for Cooperation on Agriculture (IICA) in April 2018. The Secretariat of the Economic Community of West African States has also supported the statement.

The WTO communication states that "precision biotechnology techniques, as a whole, constitute an essential tool for agricultural innovation. Their use provides farmers with access to products that increase productivity while preserving environmental sustainability."

The United States has expressed strong support for the international statement through Agriculture Secretary Sonny Perdue, who said "Precision biotechnologies such as genome editing hold great promise for both farmers and consumers around the world." In Canada, the Minister of Agriculture and Agri-Food, Lawrence MacAulay is pleased that his country joined the support. Minister MacAulay said "Today, we are sending a strong message that we stand ready to work with our global partners in support of transparent, predictable and science-based regulatory approaches to reduce potential trade disruptions and allow for the commercialization of precision biotechnology products."

For more details, read the WTO news release. Read the text of the joint statement, which is being updated as additional countries sign on, at the WTO website. The press statement issued by the United States is available at the Department of State website. The press release from Agriculture and Agri-food Canada is available here.

University students and educators from four universities have called for an expedited legislation on biotech crops under research and development in Uganda. This was during an inter-university dialogue on agricultural biotechnology, which was held at the School of Food Technology, Nutrition and Bioengineering Makerere University, Kampala on November 1, 2018. Students attending the event voiced the need for the National Biotechnology and Biosafety Bill to be passed by the parliament to enable Ugandan farmers access to improved biotech crops. 

"It is imperative that we do not just stop at discussing. Ugandan farmers urgently need this technology. Our political leaders need to pass enabling legislations to facilitate this," said Daniella Kagina, president of the Makerere University Debate Union. Dubbed as "Biotech Happy Hour", the event got many other students fired up, with compelling submissions in favor of modern biotechnology. Furthermore, the students asked that the discussion on agricultural biotechnology be taken further to the grassroots and special effort must be made to involve stakeholders who are skeptical of the technology.

The event, organized by Uganda Biosciences Information Center (UBIC), also featured a screening of the Food Evolution movie. The movie paints a picture of progress and navigates the thorny landscape of a heated global debate on agricultural biotechnology.

Over 80 participants appreciated the need to embrace modern agricultural biotechnology to address some of the most pressing challenges such as pests, diseases, and climate change. Furthermore, they pledged to engage their colleagues on this topic in a bid to further raise awareness and concretize support for the adoption of advancements in agriculture particularly in developing countries like Uganda.

In a related but separate event, farmers in Omoro district northern Uganda also called on their leaders to rally support for the delayed legislation on biotechnology. This was during a one-day biotechnology awareness workshop at Omoro District Council Hall on November 2, 2018, which brought together religious leaders, farmers, and local government representatives. The chief guest, Hon. Catherine Lamwaka, who is also Omoro district woman member of parliament promised to support the biotech bill and also present the farmers' views when the bill returns to parliament for debate this week.

For more details, contact the UBIC Coordinator at ubic.nacrri@gmail.com.

The United States Patent and Trademark Office has granted U.S. Patent Number 10,113,167, covering unique RNA guides that, when combined with the Cas9 protein, are effective at homing in on and editing genes. This RNA/protein combination act like precision-targeted gene editing scissors.

The CRISPR-Cas9 DNA-targeting complex, discovered by Jennifer Doudna, Emmanuelle Charpentier, and their teams at University of California, Berkeley and the University of Vienna, is one of the fundamental molecular technologies behind the revolutionary CRISPR-Cas9 gene editing tool.

This patent and the previous U.S. Patent Number 10,000,772 cover CRISPR-Cas9 compositions which are useful as gene editing scissors in any setting, including animal and human cells. The new patent also encompasses protein/RNA compositions that can deliver CRISPR-Cas9 into cells in two different ways: as a fully functional ribonucleoprotein (i.e., Cas9 protein complexed with RNA), and with the components encoded by DNA that is subsequently expressed and assembled inside the cell to form a functional CRISPR-Cas9 complex.

For more details, read the Berkeley News.

PennState University researchers used rice seedlings to show that the stress of hotter temperatures may trigger a response in a plant's ribonucleic acid (RNA), which is a part of a cells' genetic messaging system, to manage this change in its environment.

The researchers studied over 14,000 different RNAs to look for changes in the RNA. Thus, they searched for changes in RNA's intricately folded structures that could signal acute heat stress. Since RNA is single-stranded, unlike the double-stranded DNA, it is able to fold back on itself and form more complex folds than DNA. They exposed two-week old rice seedlings to above normal temperatures for just ten minutes and compared with the control plants.

Results showed that the folds in the RNA of plants suffering from heat stress were looser than those in the control plants. The unfolding of the mRNA, a particular type of RNA, which transfers DNA instructions to the ribosome in a cell during the protein-making process, was also found to be correlated with a loss in the abundance of mRNA, suggesting that mRNA unfolding promotes its degradation, a method that cells use to regulate which genes express and when.

According to one of the researchers, Philip Bevilacqua, the results give hints on next steps for future research into more heat and drought resistant crops.

"So, if loss of structure results in loss of abundance and if that loss of abundance is not optimal, then you could imagine that we could change the sequences of the ends of the RNA, making them more stable, and, therefore, stabilize the production of those proteins," he said.

Read more from Penn State.

The American Seed Trade Association (ASTA) expressed their approval in the statement signed by 13 countries which emphasizes their commitment to the fair, science-based treatment of evolving breeding methods such as gene editing worldwide.

According to the statement, innovations in precision biotechnology, such as gene editing, have brought the promise of major improvements in terms of the ease and precision of introducing desirable traits into agricultural organisms, as compared to other breeding methods. The statement was released in Geneva, Switzerland at the World Trade Organization (WTO) Committee on Sanitary and Phytosanitary Measures.

"This is a strong show of support by governments around the world in recognition of the necessity of continued evolution in plant breeding, and the critical role that it will play in ensuring a more sustainable and secure global food production system," said ASTA President and CEO Andrew LaVigne. "Seed is a global industry, and in light of the recent disappointing decision by the European Court of Justice, efforts such as this international statement are more important than ever in working toward the goal of global alignment on policies around agricultural innovation."

Read the news release from the American Seed Trade Association.

Scientists at the RIKEN Center for Sustainable Resource Science in Japan have found that the protein NGA1 is critical for plants to have normal responses to dehydration. In plants, dehydration response is regulated by the hormone abscisic acid (ABA). Successful rehydration requires accumulation of ABA during the early stages of dehydration, among other things. While scientists know how ABA does its work, they did not know much about how ABA begins to accumulate in response to dehydration stress. RIKEN scientist Hikaru Sato and his team screened 1,670 transgenic plant lines and performed a series of experiments to address this issue.

The team found a plant line with an overexpression of NGA with a chimeric repressor domain which resulted in reduced levels of the enzyme NCED3 during dehydration stress. This was very promising because plants need NCED3 to make ABA. The team hypothesized that NGA was a transcription factor that could control the production of NCED3, and ultimately the biosynthesis of ABA. They also found out that there is a whole family of NGA proteins, and showed that all of them bind to the region of the NCED3 gene that triggers its transcription.

The researchers then created transgenic plants for each member of the NGA family and found that NGA proteins are naturally found in different parts of plants, with different expression patterns during dehydration stress. The timing of NGA expression also varied among different plant lines which meant that not all of them function the same way during drought stress. From the mutants they created, they found that after withholding water until the plants withered, NGA1 mutants remained dried up and could not be revived through rehydration. All the other mutants could be rehydrated.

For details, read the research results published in PNAS.

The USDA Foreign Agricultural Service organized two outreach events in Jakarta and Bogor on October 24 and 25, 2018 with the objective of advancing the public understanding and acceptance of modern biotechnology applications in agriculture. The events included lectures, collaborative ideas sessions and film screening of a Dutch documentary titled Well Fed, which were attended by a diverse group of stakeholders representing academia, government, and industry. A panel discussion on biotechnology and its status in Indonesia was conducted after the film showing.

The challenges in modern agri-biotech were discussed while laying out a path for continued engagement with policymakers and the public. It was concluded that outreach on biotech research and market access is vital since Indonesia has yet to allow the commercialization of GE crops. As part of the seminar, nine influential advocates were also identified, representing the scientific, farming, and research communities to serve as "Biotech Ambassadors". Dr. Mahalecthumy Arujanan of Malaysia Biotechnology Information Center and ISAAA's Dr. Rhodora Aldemita served as resource speakers in the two-day event.

Read more from USDA FAS or contact knowledge.center@isaaa.org.

The Philippines remain as Asia's biotechnology leader, according to the USDA Foreign Agricultural Service-Global Agricultural Information Network Report on agricultural biotechnology. The Philippines is the first Asian country to allow the planting of GM crop and is moving towards developing a regulatory framework for GE animals. In 2016, the original GE plant regulations (Department of Agriculture Administrative Order No. 8) was changed to the Joint Department Circular (JDC), which led to the slow processing of biosafety applications. According to the GAIN report, such delay in approvals may affect U.S. trade as well as the country's biotech leadership status in the region.

House Deputy Speaker Sharon Garin called for the passing of House Bill 7926 or the Modern Biotechnology Act of 2018, which promoted the safe and responsible use of modern biotech and for the establishment of the Biotechnology Authority of the Philippines that will serve as the policy-making body on biotech in the country.

Read the GAIN Report from USDA FAS and the House Bill 7926.

Scientists from The University of Edinburgh have discovered a gene that could help develop disease-resistant crops. The scientists studied how plants produce nitric oxide when they are under attack from bacteria or viruses. Nitric oxide accumulates in plant cells and triggers a response from the plant's immune system.

The research team used Arabidopsis to study the genes that were triggered as nitric oxide levels rose. They found that a previously unknown gene – called SRG1 – is rapidly activated by nitric oxide and is also triggered during bacterial infection. Further studies showed that SRG1 activates the plant's defense mechanism by limiting the activity of genes that suppress the immune response. They also found that nitric oxide regulates immune response, which ensures that the plant's defense system does not over-react.

For more details, read the news article from The University of Edinburgh.

Plant communities and animals have typically performed better than monocultures. The mechanisms for this, however, have been a mystery for a long time. Biologists at the University of Zurich (UZH) have now identified the genetic cause of these mechanisms.

Two UZH researchers, Samuel Wüst and Pascal Niklaus, addressed this question by combining modern genetic and ecological approaches. They used systematic crosses of varieties of Arabidopsis plants, which were grown in pots in different combinations. A few weeks later, the researchers weighed the resulting biomass, which allowed them to compare the growth of the plants. As expected, pots with mixtures of different crosses were indeed more productive on average.

The researchers related the yield gain in mixed communities to the genetic makeup of the crosses. The genetic map they obtained helped them in identifying parts of the genome that made the combination of plants good mixed teams. They found that even the smallest genetic differences between plants were enough to increase their combined yield.

For more details, read the research news from The University of Zurich.

The Minister of Agriculture of The Netherlands, Carola Schouten, opens door to genetic modification. Minister Schouten wants to use genetic modification to make agriculture in The Netherlands more sustainable. She is currently working with companies, farmers, and Wageningen University to look at the possibilities for experimenting with gene editing CRISPR-Cas method. According to reports, Minister Schouten will send a letter to the Parliament about their progress in the coming weeks.

Wageningen University and Research said that the Minister wants to experiment with CRISPR-Cas, despite the European Court of Justice ruling. Bert Lotz, a plant researcher at the university said that it is an important signal that the Minister is giving. He added that international and national research shows that it can be done carefully and that there are major sustainability successes involved.

For more details, read the NLTmes article.

Researcher Yuan Chin from Fujian Agriculture and Forestry University in China and colleagues developed a simple protoplast isolation method that can be used for genetic transformation of pineapple. The protoplasts were isolated from cultured pineapple leaves. They tested the isolated protoplast for transient expression of proteins AtJAZ3 and AtMYC2 through PEG-mediated transformation. Results showed suitability of the method for studying protein localization and interaction. The researchers concluded that this method can also be used with genomic sequencing data to study molecular control of pineapple plant growth.

For more information, read the article in BMC Plant Methods.

The researchers expressed the gene RcPLCL1 from castor bean in Camelina via Agrobacterium-mediated transformation. To determine the oil content of the transgenic plants, the researchers extracted oil from the Camelina seeds using a modified Blight and Dyer method. Results showed increased HFA production and improved germination rate of Camelina seeds.

For more information, read the article in Frontiers in Plant Science.

Important fruit crops apple and grapevine are subjects for crop improvement by conferring biotic and abiotic stress tolerance through traditional breeding and modern biotechnology. As approvals for field testing and commercialization have been limited in most crops, researcher Yuriko Osakabe from Tokushima University in Japan and colleagues explore genome editing to genetically improve apple and grapevine.

In their article in Nature, the group of researchers described two protocols, namely, plasmid-mediated genome editing and direct delivery of CRISPR-Cas9 ribonucleoproteins for genome editing of apple and grapevine. They transformed CRISPR-Cas9 components into protoplasts and verified mutations produced by the system. Results showed high efficiency and accuracy of the system in the two fruit crops, taking 2-3 weeks for genetic transformation of the CRISPR-Cas9 system and more than three months for plant regeneration and verification of the genome edits.

For more information, read the article in Nature Protocols.

CRISPR-Cas9-mediated gene editing is enabled by the same methods as the production of genetically modified organisms. The only difference is that the transferred DNA sequence is sorted out of the plant at the end of the experiment, making the plant transgene-free. However, researcher Roman Jerala of National Institute of Chemistry in Slovenia and colleagues say that the insertion of the transgene at the earlier part of the experiment can cause unwanted mutations. Also, further expression of the transgene in the plant can cause off-target mutations. Therefore, the team proposed a method to skip transgene production in gene editing.

The team transformed Cas9 enzymes and guide RNA sequence with ribonucleoprotein into plant protoplasts using PEG-mediated transformation. Results showed up to 25 percent mutation frequency in genes FRI and PDS in the said species. They also observed positive correlation between amount of CRISPR components and mutation rate. The team aims to target other genes and study edited protoplast regeneration in the future.

For more information, read the article in Frontiers in Plant Science.

The database currently houses extensive information on gene-edited tomato, including details on genes edited using CRISPR, the name of the project, details of the transformation experiment, transformed plant variety, DNA construct used, guide RNA sequence and primers used to characterize the resulting mutations, and details about the edited plant line, such as altered sequence, zygosity, and phenotype. The database invites users to submit their own data by providing complete information about the edited plant. A comprehensive user's guide and news about CRISPR are also included in the website. 

For more information, visit the database at Plant Genome Editing Database.

Cases of Alzheimer's disease (AD), the most common form of dementia, is mostly sporadic, making it difficult for scientists to determine how the disease starts and develops. Notably, researcher Carlos Pascual-Caro from University of Extremadura in Spain and colleagues pave a step forward into finding a cure for the disease by discovering a gene linked to Alzheimer's.

In their article published in Journal of Molecular Medicine, they described the discovery of the STIM1 gene and how CRISPR helped in verifying its function and mechanism. By analyzing brain tissues from deceased AD patients and normal patients, researchers found deficiency in STIM1 in AD patients. Through CRISPR, the researchers discovered that deletion of the gene results to reduced transport of calcium ions through the plasma membrane, which causes the cell to die. 

For more information, read the articles in Synthego and Journal of Molecular Medicine.

Updated versions of the following Pocket Ks are now available for download:

Contributions of Agricultural Biotechnology in Alleviation of Poverty and Hunger
Contribution of GM Technology to the Livestock Sector
Communicating Crop Biotechnology

Other topics are also available at the ISAAA website.