The Gram-negative bacterium Agrobacterium tumefaciens causes crown gall disease on a large number of plants. The disease is characterized by the formation of tumor-like structures on stems and other organs. Tumors develop by the integration of a bacterial DNA fragment, the T-DNA, into the plant genome and by the expression of encoded genes involved in biosynthesis of plant hormones. This unique property of interkingdom gene transfer has been used for plant genetic engineering, leading to the first genetically modified plants. Despite the enormous economic importance, the exact mechanisms of T-DNA transfer still await to be unraveled. Here, we describe the agrobacteria, including A. rhizogenes, the causal agent of hairy root disease. We summarize the general properties of the tumor-inducing (Ti) and root-inducing (Ri) megaplasmids that contain the T-DNA and virulence genes implicated in T-DNA transfer. The use of agrobacteria for plant biotechnology is discussed from a historical perspective together with current improvements and their use for CRISPR/Cas9 gene editing. Finally, we report how research on Agrobacterium has led to new insights relevant to other systems, including on the type IV secretion system. We anticipate that we are only at the start of understanding Agrobacterium-related plant disease and of the development of Agrobacterium-based tools for plant genetic engineering.

The African continent has considerable potential to reap the benefits associated with modern agricultural biotechnology. Plant biotechnology and breeding represent an invaluable toolbox to face the challenges of African agriculture, such as food and nutrition security, environment protection, soil fertility, and crop adaptation to new climatic conditions. As Africa has only relatively recently adopted agricultural biotechnology, it has the opportunity to harness the immense knowledge gathered over the last two decades while avoiding some of the difficulties experienced by early adopters. High-level research and education systems together with a specific regulatory framework are critical elements in the development of sustainable biotechnology-based agriculture and industry. The more actors that are involved in Research & Development applied to nutritionally and important local crops, the faster Africa will generate its future African innovators. Here, we discuss the contribution of plant biotechnology to a transformative African agriculture that combines intensification of land productivity and environmental sustainability.

Despite years of scientific effort to develop useful and safe biotech crops, ideologies have prevailed and genetically modified (GM-)crops have still not been fully accepted by today's society. This leads one to reflect on the role of Science in society, on what makes scientists credible, and how scientists themselves understand the world we live in. While Science remains a black box for many of the uninitiated, scientists themselves are also generally less-interested in sociology or the economy, such that the coevolution of science and daily life is often frustrated by incomprehension or even disinterest on both sides.

To transform developing and least developing countries into industrialised ones, biotechnology could be deployed along the value chain, to provide support to the development of the bio-based industries in such a way to ensure sustainability of the sector and to reduce negative environmental impacts that might otherwise occur. In agribusiness development, for instance, interventions could start from inputs and agricultural mechanization, modern processing technologies, packaging of perishable products, the promotion of food safety in the processing and regulatory environment; and interventions to improve competitiveness and productivity. Worth over USD 300 billion in revenue, the role of the biotechnology goes beyond industrial growth, since it provides opportunities for progress towards many of the UN sustainable development goals (SDGs). This paper reviews the status of industrial biotechnology as it relates to inclusive and sustainable industrial development.

Neurolathyrism is a unique neurodegeneration disease caused by β-N-oxalyl-L-α, β- diaminopropionic (β-ODAP) present in grass pea seed (Lathyrus stativus L.) and its pathogenetic mechanism is unclear. This issue has become a critical restriction to take full advantage of drought-tolerant grass pea as an elite germplasm resource under climate change. We found that, in a human glioma cell line, β-ODAP treatment decreased mitochondrial membrane potential, leading to outside release and overfall of Ca2+ from mitochondria to cellular matrix. Increased Ca2+ in cellular matrix activated the pathway of ECM, and brought about the overexpression of β1 integrin on cytomembrane surface and the phosphorylation of focal adhesion kinase (FAK). The formation of high concentration of FA units on the cell microfilaments further induced overexpression of paxillin, and then inhibited cytoskeleton polymerization. This phenomenon turned to cause serious cell microfilaments distortion and ultimately cytoskeleton collapse. We also conducted qRT-PCR verification on RNA-sequence data using 8 randomly chosen genes of pathway enrichment, and confirmed that the data was statistically reliable. For the first time, we proposed a relatively complete signal pathway to neurolathyrism. This work would help open a new window to cure neurolathyrism, and fully utilize grass pea germplasm resource under climate change.

RNA interference (RNAi) technology can potentially serve as a suitable strategy to control the African sweet potato weevil Cylas puncticollis (SPW), which is a critical pest in sub-Saharan Africa. Important prerequisites are required to use RNAi in pest control, such as the presence of an efficient RNAi response and the identification of suitable target genes. Here we evaluated the toxicity of dsRNAs targeting essential genes by injection and oral feeding in SPW. In injection assays, 12 of 24 dsRNAs were as toxic as the one targeting Snf7, a gene used commercially against Diabrotica virgifera virgifera. Three dsRNAs with high insecticidal activity were then chosen for oral feeding experiments. The data confirmed that oral delivery can elicit a significant toxicity, albeit lower compared with injection. Subsequently, ex vivo assays revealed that dsRNA is affected by degradation in the SPW digestive system, possibly explaining the lower RNAi effect by oral ingestion. We conclude that the full potential of RNAi in SPW is affected by the presence of nucleases. Therefore, for future application in crop protection, it is necessary constantly to provide new dsRNA and/or protect it against possible degradation in order to obtain a higher RNAi efficacy. © 2016 Society of Chemical Industry.

Genetic polymorphisms in coding genes play an important role when using mouse inbred strains as research models. They have been shown to influence research results, explain phenotypical differences between inbred strains, and increase the amount of interesting gene variants present in the many available inbred lines. SPRET/Ei is an inbred strain derived from Mus spretus that has ∼1% sequence difference with the C57BL/6J reference genome. We obtained a listing of all SNPs and insertions/deletions (indels) present in SPRET/Ei from the Mouse Genomes Project (Wellcome Trust Sanger Institute) and processed these data to obtain an overview of all transcripts having nonsynonymous coding sequence variants. We identified 8,883 unique variants affecting 10,096 different transcripts from 6,328 protein-coding genes, which is about 28% of all coding genes. Because only a subset of these variants results in drastic changes in proteins, we focused on variations that are nonsense mutations that ultimately resulted in a gain of a stop codon. These genes were identified by in silico changing the C57BL/6J coding sequences to the SPRET/Ei sequences, converting them to amino acid (AA) sequences, and comparing the AA sequences. All variants and transcripts affected were also stored in a database, which can be browsed using a SPRET/Ei M. spretus variants web tool (www.spretus.org), including a manual. We validated the tool by demonstrating the loss of function of three proteins predicted to be severely truncated, namely Fas, IRAK2, and IFNγR1.

When we discovered that crown gall induction on plants by Agrobacterium tumefaciens is a natural event of genetic engineering, we were convinced that this was the dawn of a new era for plant science. Now, more than 30 years later, I remain overawed by how far and how rapidly we progressed with our knowledge of the molecular basis of plant growth, development, stress resistance, flowering, and ecological adaptation, thanks to the gene engineering technology. I am impressed, but also frustrated by the difficulties of applying this knowledge to improve crops and globally develop a sustainable and improved high-yielding agriculture. Now that gene engineering has become so efficient, I had hoped that thousands of teams, all over the world, would work on improving our major food crops, help domesticate new ones, and succeed in doubling or tripling biomass yields in industrial crops. We live in a world where more than a billion people are hungry or starving, while the last areas of tropical forest and wild nature are disappearing. We urgently need a better supply of raw material for our chemical industry because petroleum-based products pollute the environment and are limited in supply. Why could this new technology not bring the solutions to these challenges? Why has this not happened yet; what did we do wrong?

The excitatory amino acid L-β-N-oxalyl-α,β-diaminopropionic acid (L-β-ODAP) in Lathyrus sativus L. is proposed as the causative agent of the neurodegenerative disease neurolathyrism. We investigated the effect of L-β-ODAP on [Ca2+]i handling, redox homeostasis, and cell death in rat spinal motor neurons. L-β-ODAP and L-glutamate triggered [Ca2+]i transients, which were inhibited by the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor blockers; 2,3-dioxo-6-nitro-1,2,3, 4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide and 1-naphthyl acetylspermine, the latter specifically blocking Ca2+-permeable α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors. In addition, 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide, and to a lesser extent 1-naphthyl acetylspermine, protected the neurons against cell death induced by L-β-ODAP or L-glutamate. Methionine and cysteine were also protective against neuronal cell death. We conclude that deregulation of [Ca2+]i homeostasis and oxidative stress contribute to motor neuron cell death in neurolathyrism.