Cisgenesis: an important sub-invention for traditional plant breeding companies

Modern plant breeding is highly dependent on new technologies to master future problems. More traits have to be combined, frequently originating from wild species. Traditional breeding is connected with linkage drag problems. The crop plant itself and its crossable species represent the traditional breeders gene pool. GM-breeding is a new way of improving existing varieties. Transgenes originate from non-crossable species and are representing a new gene pool. For release of GM-plants into the environment and onto the market in Europe Directive 2001/18/EC has been developed, primarily based on GM-technology and not on gene source. In society, opposition against GM crops is complicating the implementation of GM crops. In this paper, it is shown that not only transgenes, representing a new gene pool but also cisgenes and intragenes are available, representing the breeders gene pool. Cisgenes are natural genes and intragenes are composed of functional parts of natural genes from the crop plant itself or from crossable species. Cisgenesis is the combined use of only cisgenes with marker-free transformation, mimicking linkage drag free introgression breeding in one step. Therefore, cisgenesis is a new sub-invention in the traditional breeding field and indicates the need for reconsideration of GM Directives. Inventions are frequently containing not only hardware elements, but also software and orgware elements. For cisgenesis it is foreseen that the technical (hardware) and bioinformatic (software) elements will develop smoothly, but that implementation in society is highly dependent on acceptance and regulations (orgware). It could be made in a step by step approach by specific crop-gene derogations from the Directive, followed by adding cisgenesis to annex 1b of Directive 2001/18/EC for exemption. At present GM crops can only be introduced by large companies. An open innovation approach for cisgenesis by public private partnership including traditional SMEs has been discussed. Cisgenesis has been exemplified for resistance breeding of potato to Phytophthtora infestans

Cisgenesis, a new tool for traditional plant breeding, should be exempted from the regulation on genetically modified organisms in a step by step approach

Modern potato breeding requires over 100,000 seedlings per new variety. Main reasons are (1) the increasing number of traits that have to be combined in this tetraploid vegetatively propagated crop, and (2) an increasing number of traits (e.g., resistance to biotic stress) originates from wild species. Pre-breeding by introgression or induced translocation is an expensive way of transferring single traits (such as R-genes, coding for resistance to biotic stress) to the cultivated plant. The most important obstacle is simultaneous transfer of undesired neighbouring alien alleles as linkage drag. Stacking several genes from different wild sources is increasing this linkage drag problem tremendously. Biotechnology has enabled transformation of alien genes into the plant. Initially, transgenes were originating mainly from microorganisms, viruses or non-crossable plant species, or they were chimeric. Moreover, selection markers coding for antibiotic resistance or herbicide resistance were needed. Transgenes are a new gene source for plant breeding and, therefore, additional regulations like the EU Directive 2001/18/EC were developed. Because of a strong opposition against genetic modification of plants in Europe, the application of this Directive is strict, very expensive, hampering the introduction of genetically modified (GM) crops and the use of this technology by small and medium-sized enterprises (SMEs). Currently, GM crops are almost the exclusive domain of multinationals. Meanwhile, not only transgenes but also natural genes from the plant species itself or from crossable plant species, called cisgenes, are available and the alien selection genes can be avoided in the end product. This opens the way for cisgenic crops without alien genes. The existing EU directive for GM organisms is not designed for this new development. The cisgenes belong to the existing breeders¿ gene pool. The use of this classical gene pool has been regulated already in agreements regarding breeders¿ rights. We are proposing a step by step approach starting with a crop and gene specific derogation and monitoring towards a general exemption of cisgenic plants from the Directive. Two examples, i.e. development of cisgenic potato for resistance to Phytophthora infestans and cisgenic apple for resistance to Venturia inaequalis are discussed shortly for illustration of the importance of cisgenesis as a new tool for traditional plant breeding. Cisgenesis is simplifying introgression and induced translocation breeding tremendously and is highly recommended for SMEs and developing countrie

QND measurement of a superconducting qubit in the weakly projective regime

Quantum state detectors based on switching of hysteretic Josephson junctions biased close to their critical current are simple to use but have strong back-action. We show that the back-action of a DC-switching detector can be considerably reduced by limiting the switching voltage and using a fast cryogenic amplifier, such that a single readout can be completed within 25 ns at a repetition rate of 1 MHz without loss of contrast. Based on a sequence of two successive readouts we show that the measurement has a clear quantum non-demolition character, with a QND fidelity of 75 %.Comment: submitted to PR

First attempts of linking modelling, Postharvest behaviour and Melon Genetics

The onset of climacteric is associated with the end of melon fruit shelf-life. The aim of this research was to develop practical and applicable models of fruit ripening changes (hardness, moisture loss) also able to discriminate between climacteric and non-climacteric behaviour. The decrease in firmness was measured non-destructively by flat-plate compression; moisture loss was measured by weight loss. A set of 13-15 near-isogenic lines (NILs) derived from the climacteric line SC3-5 was used to verify the relationship among the climacteric behaviour and ripening related changes (weight loss, softening and color) during two consecutive seasons. The biological variance models for moisture loss and firmness followed a simple exponential behaviour that explained more than 90% of the total variance. Results of the analyses using these models could not be linked to properties of near-isogenic lines like climacteric behaviour, ethylene production or skin thickness. The results suggest that the phenotype is more important than genotype, when considering mean values. These results seem to suggest that relations may exist between the different processes and properties of NILs on an individual basis, not on mean values

Development of cisgenic apples with durable resistance to apple scab

Most of the apple (Malus × domestica) growers are facing serious disease problems with apple scab which is caused by the fungus Venturia inaequalis. Developing a resistant variety in apple through classical breeding is very slow and inefficient. So, we aim at improving existing apple varieties through a new concept called “cisgenesis” which saves time and effort compared to classical breeding. Malus floribunda proved to be a good source of natural scab resistance genes. The genes HcrVf1 and HcrVf2, consisting of promoter, coding and terminator sequences in their natural configuration, were isolated from Malus floribunda and cloned into the binary vector pMF1. Apple cv. ‘Gala’ was transformed with pMF1 containing HcrVf1 and HcrVf2, individually or in combination. pMF1 can be used to obtain marker-free plants by recombinase-based excision of a fragment carrying undesired gene sequences, such as antibiotic-selection marker genes, leaving behind only the gene(s)-of-interest and one recombination site. Using this vector it is therefore possible to stack several genes by retransformation using the same selection procedure. In order to obtain durable resistance, we have the intention to combine different resistance genes from Malus either by stacking them one by one or by introducing them all together in one T-DNA. Performance of all different types of cisgenic plants will be evaluated by monitoring scab resistance levels phenotypically and by determining gene expression profiles through quantitative RT-PC

Water loss in horticultural products. Modelling, data analysis and theoretical considerations

The water loss of individual fruit (melon, plum and mandarin) was analysed using the traditional diffusion based approach and a kinetic approach. Applying simple non linear regression, both approaches are the same, resulting in a quite acceptable analysis. However, by applying mixed effects non linear regression analysis, explicitly including the variation over the individuals, the kinetic approach was found to reflect the processes occurring during mass loss better than the diffusion approach. All the variation between the individuals in a batch could be attributed to the initial mass or size of the individuals. The fraction of the fruit mass that is available for transpiration is the key item in the water loss process, rather than the skin resistance and fruit area. Obtained explained parts are well over 99%

Inner topological structure of Hopf invariant

In light of $\phi$-mapping topological current theory, the inner topological structure of Hopf invariant is investigated. It is revealed that Hopf invariant is just the winding number of Gauss mapping. According to the inner structure of topological current, a precise expression for Hopf invariant is also presented. It is the total sum of all the self-linking and all the linking numbers of the knot family.Comment: 13pages, no figure. Accepted by J.Math.Phy

Incremental Distance Transforms (IDT)

A new generic scheme for incremental implementations of distance transforms (DT) is presented: Incremental Distance Transforms (IDT). This scheme is applied on the cityblock, Chamfer, and three recent exact Euclidean DT (E2DT). A benchmark shows that for all five DT, the incremental implementation results in a significant speedup: 3.4×−10×. However, significant differences (i.e., up to 12.5×) among the DT remain present. The FEED transform, one of the recent E2DT, even showed to be faster than both city-block and Chamfer DT. So, through a very efficient incremental processing scheme for DT, a relief is found for E2DT’s computational burden