Field testing and exploitation of genetically modified cassava with low-amylose or amylose-free starch in Indonesia

The development and testing in the field of genetically modified -so called- orphan crops like cassava in tropical countries is still in its infancy, despite the fact that cassava is not only used for food and feed but is also an important industrial crop. As traditional breeding of cassava is difficult (allodiploid, vegetatively propagated, outbreeding species) it is an ideal crop for improvement through genetic modification. We here report on the results of production and field testing of genetically modified low-amylose transformants of commercial cassava variety Adira4 in Indonesia. Twenty four transformants were produced and selected in the Netherlands based on phenotypic and molecular analyses. Nodal cuttings of these plants were sent to Indonesia where they were grown under biosafety conditions. After two screenhouse tests 15 transformants remained for a field trial. The tuberous root yield of 10 transformants was not significantly different from the control. Starch from transformants in which amylose was very low or absent showed all physical and rheological properties as expected from amylose-free cassava starch. The improved functionality of the starch was shown for an adipate acetate starch which was made into a tomato sauce. This is the first account of a field trial with transgenic cassava which shows that by using genetic modification it is possible to obtain low-amylose cassava plants with commercial potential with good root yield and starch quality.     

能源木薯高淀粉抗逆分子育种研究进展与展望

木薯(Manihot esculenta Crantz)是全球重要的粮食作物,也是我国非粮生物质能源发展的主要原材料。长期以来,传统杂交育种是木薯新品种培育的主要手段。随着全球生态的变化和木薯产业发展的推进,需要加速培育抗逆能力强、高淀粉的木薯新品种,因此,利用基因工程针对特定性状开展品种创新表现出巨大的潜力。随着组学技术的发展,在木薯基础研究领域,特别是针对储藏根发育、淀粉富集、逆境响应与调控等方面的研究逐步深入。强化木薯基础理论研究和发展应用技术,对推动能源木薯的产业化发展具有重要意义。     

木薯分子育种中若干基本科学问题的思考与研究进展

木薯作为全球重要的薯类作物,既是热带地区粮食安全的保障,也是重要的淀粉工业原材料,保障其稳产、高产、优质一直是育种家不变的研究主题．当前,木薯品种选育正处在从杂交育种转向分子育种的发展阶段,深入解析木薯特有的经济性状和生物学特点是利用生物技术进行遗传改良的基础．不同于谷物类作物,木薯光合同化物的转运和库源分配的调控机制必有其独特之处;同时,储藏根的“库容”直接影响其产量．作为热带作物,了解木薯对低温和干旱的响应可为改良其抗逆境能力提供理论依据．不同于其他薯类作物,木薯储藏根特有的“采后生理性变质”问题亟待解决,其发生和调控机制的解析对延长木薯货架期意义重大．随着分子生物学的发展,针对上述各方面的研究日益深入,不仅激发了感兴趣的公众对这些问题的认知和思考,也激励了科研人员不断努力寻找解析相关机制的方法,为最终通过分子育种手段改良木薯提供思路和技术方案,揭开木薯的层层“面纱”,推动木薯分子育种的发展．     

植物分子育种技术在改良木薯种质方面的应用

木薯是热带、亚热带地区的重要粮食作物和经济作物。培育出生产性状更加优良的木薯品种,是推进木薯产业更快更好发展的重要基础。分子育种技术在培育优良木薯品种方面具有传统育种技术不可比拟的优势。该文介绍了近年来在提高植物抗寒性与病虫害抗性、降低氰苷含量、提高淀粉含量及组成、改变储存物种类、防止木薯收获后变质等方面的研究进展,以便在这些研究进展的基础上,利用植物分子育种技术加快获得具有抗逆能力提高、品质改良、产量增加、耐储藏等优良特性的木薯新品种。     

Cassava stems: a new resource to increase food and fuel production

Given the growing global population, mankind must find new ways to lower competition for land between food and fuel production. Our findings for cassava suggest that this important crop can substantially increase the combined production of both food and fuel. Cassava stems have previously been overlooked in starch and energy production. These food‐crop residues contain about 30% starch (dry mass) mostly in the xylem rather than phloem tissue. Up to 15% starch of the stem dry mass can be extracted using simple water‐based techniques, potentially leading to an 87% increase in global cassava starch production. The integration of biofuel production, using residues and wastewater from starch extraction, may bring added value. The cassava roots on which biofuels and other products are based can be replaced by cassava stems without land use expansion, making root starch available as food for additional 30 million people today.     

Genetic diversity of traditional South American Landraces of Cassava (Manihot esculenta Crantz): an analysis using microsatellites

The extent and structure of the genetic variability of traditional varieties of cassava (Manihot esculenta Crantz) have been little documented, despite considerable evidence for this crop? great varietal diversity in traditional agroecosystems. We used microsatellite markers to assess the genetic structure of traditional landraces of sweet and bitter cassava collected from five South American sites. As reference, we used a sample of 38 accessions from a world collection of cultivated cassava. For a total of 10 loci examined, we found 15 alleles that were not represented in this sample. Ten of these had been previously detected in wild Manihot species. The geographical structure of genetic variability was weak, but the genetic differentiation between bitter and sweet landraces was significant, suggesting that each form had evolved separately after domestication. Our results showed that traditional landraces form an important source of genetic diversity and merit more attention from managers of crop genetic resources.     

Cassava Breeding: Current Status, Bottlenecks and the Potential of Biotechnology Tools

Cassava is an important energy source in the diets of millions of people in tropical and subtropical regions of the world. It is a key subsistence crop, and its industrial uses are steadily growing. In spite of its economic and social relevance, relatively little investment has been made for research on cassava. However, conventional breeding resulted in more stable production through enhanced tolerance to biotic and abiotic stresses; increased productivity, both in fresh root production and increased dry matter content; and, more recently, improvements in qualitative traits such as starch quality and increased carotenoids content. The inbreeding of cassava has been identified as a key step for more efficient genetic improvement of the crop, therefore, research is underway to develop protocol(s) for the production of doubled haploids. Marker-assisted selection has been successfully applied to cassava, but in a more modest scale compared with other crops. More support and emphasis is needed on practical applications of molecular marker technology in cassava improvement. The availability of more efficient genotyping approaches and the cassava genome sequence promise to increase the impact of biotechnology tools on cassava improvement. Efficient and reliable phenotyping of cassava remains a challenging goal to achieve in the near future.     

Cassava breeding: opportunities and challenges

Although cassava is a major food crop, its scientific breeding began only recently compared with other crops. Significant progress has been achieved, particularly in Asia where cassava is used mainly for industrial processes and no major biotic constraints affect its productivity. Cassava breeding faces several limitations that need to be addressed. The heterozygous nature of the crop and parental lines used to generate new segregating progenies makes it difficult to identify parents with good breeding values. Breeding so far has been mainly based on a mass phenotypic recurrent selection. There is very little knowledge on the inheritance of traits of agronomic relevance. Several approaches have been taken to overcome the constraints in the current methodologies for the genetic improvement of cassava. Evaluations at early stages of selection allow for estimates of general combining ability effect or breeding values of parental lines. Inbreeding by sequential self-pollination facilitates the identification of useful recessive traits, either already present in the Manihot gene pool or induced by mutagenesis.     

Improved Cassava Starch by Antisense Inhibition of Granule-bound Starch Synthase I

Cassava is a poor man's crop which is mainly grown as a subsistence crop in many developing countries. Its commercial use was first as animal feed (also known as tapioca), but has shifted since the late sixties to a source of native starch. The availability of native starches, which on the one hand do not require substantial chemical derivatisation and on the other hand have improved properties, would make cassava also for small farmers a potentially attractive cash crop. Since breeding is difficult in this polyploid, vegetatively propagated, crop a transgenic approach would be ideal to improve certain characteristics. We have created a cassava genotype producing amylose-free starch by genetic modification. The absence of amylose increased the clarity and stability of gels made with the transgenic starch, without requiring treatment with environment-unfriendly chemicals such as epoxides (propylene oxide, ethylene oxide) and acetic anhydride, which are normally used to improve stability. The amylose-free starch showed no changes in particle size distribution, chain length distribution or phosphorous content when compared to amylose-containing starch, but the granule melting temperature was increased by almost 2°C. Furthermore, the amylose-free cassava starch shows enhanced clarity and stability properties. These improved functionalities are desired in technical applications in paper and textile manufacturing, but also in the food industry for the production of sauces, dairy products and noodles.     

Identification,validation and high-throughput genotyping of transcribed gene SNPs in cassava

The availability of genomic resources can facilitate progress in plant breeding through the application of advanced molecular technologies for crop improvement. This is particularly important in the case of less researched crops such as cassava, a staple and food security crop for more than 800 million people. Here, expressed sequence tags (ESTs) were generated from five drought stressed and well-watered cassava varieties. Two cDNA libraries were developed: one from root tissue (CASR), the other from leaf, stem and stem meristem tissue (CASL). Sequencing generated 706 contigs and 3,430 singletons. These sequences were combined with those from two other EST sequencing initiatives and filtered based on the sequence quality. Quality sequences were aligned using CAP3 and embedded in a Windows browser called HarvEST:Cassava which is made available. HarvEST:Cassava consists of a Unigene set of 22,903 quality sequences. A total of 2,954 putative SNPs were identified. Of these 1,536 SNPs from 1,170 contigs and 53 cassava genotypes were selected for SNP validation using Illumina’s GoldenGate assay. As a result 1,190 SNPs were validated technically and biologically. The location of validated SNPs on scaffolds of the cassava genome sequence (v.4.1) is provided. A diversity assessment of 53 cassava varieties reveals some sub-structure based on the geographical origin, greater diversity in the Americas as opposed to Africa, and similar levels of diversity in West Africa and southern, eastern and central Africa. The resources presented allow for improved genetic dissection of economically important traits and the application of modern genomics-based approaches to cassava breeding and conservation.     

RIKEN Cassava Initiative: Establishment of a Cassava Functional Genomics Platform

Cassava (Manihot esculenta Crantz) is an important tropical root crop that provides food security and income generation for many farmers. Cassava ranks as the third most important source of calories in the tropics and provides sustenance to 800 million people worldwide. Cassava is also an efficient source of modified starch and bioethanol. In spite of the many advantages and high potential of cassava, the research community lacks functional genomic resources. Therefore, RIKEN has been working to establish a cassava functional genomics platform, which includes full-length cDNAs, DNA microarrays, transformation capabilities, and a searchable database in collaboration with the International Center for Tropical Agriculture (CIAT) of Colombia and Mahidol University of Thailand. In this review, we summarize the present status and future perspectives of our cassava functional genomic approaches.     

Cassava: constraints to production and the transfer of biotechnology to African laboratories

Knowledge and technology transfer to African institutes is an important objective to help achieve the United Nations Millennium Development Goals. Plant biotechnology in particular enables innovative advances in agriculture and industry, offering new prospects to promote the integration and dissemination of improved crops and their derivatives from developing countries into local markets and the global economy. There is also the need to broaden our knowledge and understanding of cassava as a staple food crop. Cassava (Manihot esculenta Crantz) is a vital source of calories for approximately 500 million people living in developing countries. Unfortunately, it is subject to numerous biotic and abiotic stresses that impact on production, consumption, marketability and also local and country economics. To date, improvements to cassava have been led via conventional plant breeding programmes, but with advances in molecular-assisted breeding and plant biotechnology new tools are being developed to hasten the generation of improved farmer-preferred cultivars. In this review, we report on the current constraints to cassava production and knowledge acquisition in Africa, including a case study discussing the opportunities and challenges of a technology transfer programme established between the Mikocheni Agricultural Research Institute in Tanzania and Europe-based researchers. The establishment of cassava biotechnology platform(s) should promote research capabilities in African institutions and allow scientists autonomy to adapt cassava to suit local agro-ecosystems, ultimately serving to develop a sustainable biotechnology infrastructure in African countries.     

Is Cassava the Answer to African Climate Change Adaptation?

This paper examines the impacts of climate change on cassava production in Africa, and questions whether cassava can play an important role in climate change adaptation. First, we examine the impacts that climate change will likely have on cassava itself, and on other important staple food crops for Africa including maize, millets, sorghum, banana, and beans based on projections to 2030. Results indicate that cassava is actually positively impacted in many areas of Africa, with ?3.7% to +17.5% changes in climate suitability across the continent. Conversely, for other major food staples, we found that they are all projected to experience negative impacts, with the greatest impacts for beans (?16%?±?8.8), potato (?14.7?±?8.2), banana (?2.5%?±?4.9), and sorghum (?2.66%?±?6.45). We then examined the likely challenges that cassava will face from pests and diseases through the use of ecological niche modeling for cassava mosaic disease, whitefly, brown streak disease and cassava mealybug. The findings show that the geographic distribution of these pests and diseases are projected to change, with both new areas opening up and areas where the pests and diseases are likely to leave or reduce in pressure. We finish the paper by looking at the abiotic traits of priority for crop adaptation for a 2030 world, showing that greater drought tolerance could bring some benefits in all areas of Africa, and that cold tolerance in Southern Africa will continue to be a constraint for cassava despite a warmer 2030 world, hence breeding needs to keep a focus on this trait. Importantly, heat tolerance was not found to be a major priority for crop improvement in cassava in the whole of Africa, but only in localized pockets of West Africa and the Sahel. The paper concludes that cassava is potentially highly resilient to future climatic changes and could provide Africa with options for adaptation whilst other major food staples face challenges.     

Cassava brown streak disease: historical timeline,current knowledge and future prospects

Cassava is the second most important staple food crop in terms of per capita calories consumed in Africa and holds potential for climate change adaptation. Unfortunately, productivity in East and Central Africa is severely constrained by two viral diseases: cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). CBSD was first reported in 1936 from northeast Tanzania. For approximately 70 years, CBSD was restricted to coastal East Africa and so had a relatively low impact on food security compared with CMD. However, at the turn of the 21st century, CBSD re‐emerged further inland, in areas around Lake Victoria, and it has since spread through many East and Central African countries, causing high yield losses and jeopardizing the food security of subsistence farmers. This recent re‐emergence has attracted intense scientific interest, with studies shedding light on CBSD viral epidemiology, sequence diversity, host interactions and potential sources of resistance within the cassava genome. This review reflects on 80 years of CBSD research history (1936–2016) with a timeline of key events. We provide insights into current CBSD knowledge, management efforts and future prospects for improved understanding needed to underpin effective control and mitigation of impacts on food security.     

Cassava about‐FACE: Greater than expected yield stimulation of cassava (Manihot esculenta) by future CO2 levels

Globally, cassava is the second most important root crop after potatoes and the fifth most important crop overall in terms of human caloric intake. In addition to its growing global importance for feed, fuel, and starch, cassava has long been vital to food security in Sub‐Saharan Africa. Climate change is expected to have its most severe impact on crops in food insecure regions, yet little is known about how cassava productivity will respond to climate change. The most important driver of climate change is globally increasing atmospheric CO₂ concentration ([CO₂]). However, the potential for cassava to enhance food security in an elevated [CO₂] world is uncertain as greenhouse and open top chamber (OTC) study reports are ambiguous. Studies have yielded misleading results in the past regarding the effect of elevated [CO₂] on crop productivity, particularly in cases where pots restricted sink growth. To resolve these conflicting results, we compare the response of cassava to growth at ambient (ca. 385 ppm) and elevated [CO₂] (585 ppm) under field conditions and fully open air [CO₂] elevation. After three and half months of growth at elevated [CO₂], above ground biomass was 30% greater and cassava root tuber dry mass increased over 100% (fresh weight increased 89%). High photosynthetic rates and photosynthetic stimulation by elevated [CO₂], larger canopies, and a large sink capacity all contributed to cassava's growth and yield stimulation. Cassava exhibited photosynthetic acclimation via decreased Rubisco capacity early in the season prior to root tuber initiation when sink capacity was smaller. Importantly, and in contrast to a greenhouse study, we found no evidence of increased leaf N or total cyanide concentration in elevated [CO₂]. All of our results are consistent with theoretical expectations; however, the magnitude of the yield increase reported here surpasses all other C₃ crops and thus exceeds expectations.     

Comparative Transcriptomic Analysis of Storage Roots in Cassava During Postharvest Physiological Deterioration

Plant Molecular Biology Reporter - Cassava is an important starchy and food crop; however, the commercial value of cassava is seriously constrained by postharvest physiological deterioration (PPD)....     

Informal "seed" systems and the management of gene flow in traditional agroecosystems: the case of cassava in Cauca, Colombia

Our ability to manage gene flow within traditional agroecosystems and their repercussions requires understanding the biology of crops, including farming practices' role in crop ecology. That these practices' effects on crop population genetics have not been quantified bespeaks lack of an appropriate analytical framework. We use a model that construes seed-management practices as part of a crop's demography to describe the dynamics of cassava (Manihot esculenta Crantz) in Cauca, Colombia. We quantify several management practices for cassava--the first estimates of their kind for a vegetatively-propagated crop--describe their demographic repercussions, and compare them to those of maize, a sexually-reproduced grain crop. We discuss the implications for gene flow, the conservation of cassava diversity, and the biosafety of vegetatively-propagated crops in centers of diversity. Cassava populations are surprisingly open and dynamic: farmers exchange germplasm across localities, particularly improved varieties, and distribute it among neighbors at extremely high rates vis-à-vis maize. This implies that a large portion of cassava populations consists of non-local germplasm, often grown in mixed stands with local varieties. Gene flow from this germplasm into local seed banks and gene pools via pollen has been documented, but its extent remains uncertain. In sum, cassava's biology and vegetative propagation might facilitate pre-release confinement of genetically-modified varieties, as expected, but simultaneously contribute to their diffusion across traditional agroecosystems if released. Genetically-modified cassava is unlikely to displace landraces or compromise their diversity; but rapid diffusion of improved germplasm and subsequent incorporation into cassava landraces, seed banks or wild populations could obstruct the tracking and eradication of deleterious transgenes. Attempts to regulate traditional farming practices to reduce the risks could compromise cassava populations' adaptive potential and ultimately prove ineffectual.     

Production of transgenic rice with agronomically useful genes: an assessment

Rice is the most important food crop in tropical and subtropical regions of the world. Yield enhancement to increase rice production is one of the essential strategies to meet the demand for food of the growing population. Both abiotic and biotic features limit adversely the productivity of rice growing areas. Conventional breeding has been an effective means for developing high yielding varieties, however; it is associated with its own limitations. It is envisaged that recent trends in biotechnology can contribute to the agronomic improvement of rice in terms of yield and nutritional quality as a supplement to traditional breeding methods. Genetic transformation of rice has demonstrated numerous important opportunities resulting in the genetic improvement of existing elite rice varieties and production of new plant types. Significant advances have been made in the genetic engineering of rice since the first transgenic rice plant production in the late 1980s. Several gene transfer protocols have been employed successfully for the introduction of foreign genes to rice. In more than 60 rice cultivars belonging to indica, japonica, javanica, and elite African cultivars, the protocol has been standardized for transgenic rice production. Selection and use of appropriate promoters, selectable markers, and reporter genes has been helpful for development of efficient protocols for transgenic rice in a number of rice cultivars. The present review is an attempt to assess the current state of development in transgenic rice for the transfer of agronomically useful genes, emphasizing the application and future prospects of transgenic rice production for the genetic improvement of this food crop.