Self-pollinated Species Research Articles

Nutritional Composition, Fatty Acid Content, and Mineral Content of Nine Sorghum (Sorghum bicolor) Inbred Varieties.

Sorghum is a self-pollinating species belonging to the Poaceae family characterized by a resistance to drought higher than that of corn. Sorghum (Sorghum bicolor L. Moench) has been grown for centuries as a food crop in tropical areas where it has an increasing importance, particularly as a cereal option for people with celiac disease. Over the past fifty years, food-grade varieties and hybrid seeds with white pericarp have been developed, particularly in the United States, to maximize sorghum food quality. Nutrient composition, including moisture, protein, carbohydrates, dietary fiber, fat content, fatty acid composition, and mineral content, was determined for nine inbred varieties with a stabilized food-grade sorghum genotype selected in the USA and grown under typical Mediterranean conditions. Differences in these nutritional components were observed among the varieties considered. Notable differences were found for monounsaturated and polyunsaturated fats, while saturated fatty acids were similar in all varieties. Oleic, linoleic, and palmitic acids were the most abundant fatty acids in all nine lines. Differences were also noted in mineral content, particularly for K, Mg, Al, Mn, Fe, Cu, Zn, and Ba. Enzyme-linked immunosorbent assays (ELISAs) demonstrated the absence of gliadin-like peptides in all the sorghum varieties analyzed, confirming, thus, that these analyzed varieties are safe for consumption by celiac patients. Knowledge of the nutritional values of sorghum lines is relevant for breeding programs devoted to sorghum nutritional content and for beneficial properties to human health.

Effective dispersal patterns in prairie plant species across human-modified landscapes.

Effective dispersal among plant populations is dependent on vector behaviour, landscape features and availability of adequate habitats. To capture landscape feature effects on dispersal, studies must be conducted at scales reflecting single-generation dispersal events (mesoscale). Many studies are conducted at large scales where genetic differentiation is due to dispersal occurring over multiple generations, making it difficult to interpret the effects of specific landscape features on vector behaviour. Genetic structure at the mesoscale may be determined by ecological and evolutionary processes, such as the consequences of vector behaviour on patterns of gene flow. We used chloroplast haplotypes and nuclear genome SNP surveys to identify landscape features influencing seed and pollen dispersal at a mesoscale within the Rogue River Valley in southern Oregon. We evaluated biotic and abiotic vector behaviour by contrasting two annual species with differing dispersal mechanisms; Achyrachaena mollis (Asteraceae) is a self-pollinating and anemochoric species, and Plectritis congesta (Caprifoliaceae) is biotically pollinated with barochoric seeds. Using landscape genetics methods, we identified features of the study region that conduct or restrict dispersal. We found chloroplast haplotypes were indicative of historic patterns of gene flow prior to human modification of landscapes. Seed dispersal of A. mollis was best supported by models of isolation by distance, while seed-driven gene flow of P. congesta was determined by the distribution of preserved natural spaces and quality habitat. Nuclear genetic structure was driven by both pollen and seed dispersal, and both species responded to contemporary landscape changes, such as urban and agricultural conversion, and habitat availability.

Genetic diversity of Sinapis arvensis L. (wild mustard) in Türkiye determined by microsatellite markers

Wild mustard (Sinapis arvensis L.) is a self-fertilizing weed species that exerts negative impacts on wheat production and herbicides are intensively used to manage it. Cross-fertilization may lead to genetic differentiation in this species. Therefore, this study investigated genetic diversity among wild mustard populations collected from wheat fields across various regions in Türkiye. Genetic variation was evaluated using 5 simple sequence repeat (SSR) markers in populations collected from 30 different locations. Populations were analyzed using UPGMA (unweighted pair group method with arithmetic mean) and principal component analysis (PCA). The mean genetic diversity (GD) and polymorphism information content (PIC) values were 0.752 and 0.844, respectively. High genetic variability was recorded among populations within geographic locations. The populations were categorized into two major groups by UPGMA. There was no apparent geographic isolation among tested populations, which displayed a high degree of variability. The primary source of this variability is thought to be the adaptability of wild mustard seeds dispersed through various methods across diverse locations. Despite being a predominantly self-pollinating species, wild mustard may also employ some cross-pollination mechanisms. In conclusion, SSR markers proved useful in determining genetic diversity in outcrossing species, especially where no prior genotypic information is available. The study suggests that genetic diversity is maintained in wild mustard populations even with rotational farming practices and intensive use of herbicides.

Hybrid Mimulus flowers attract a new pollinator.

Hybridization is common in flowering plants and is believed to be an important force driving adaptation and speciation. The flowers of hybrids often exhibit new trait combinations, which, theoretically, could attract new species of pollinators. In this study, we found that the hybrids between a hummingbird-pollinated species Mimulus cardinalis and a self-pollinated species Mimulus parishii attract bumblebees (Bombus impatiens), a pollinator not attracted to either of the progenitor species. This novel attraction is explained by new combinations of floral traits in hybrids, including, most importantly, petal color, in addition to nectar concentration and corolla size. To understand how petal color variation is perceived by bumblebees, we performed reflectance spectroscopy and multispectral imaging to model the flower appearance in bee vision. This analysis showed that color variation would impact the ease of detection. We also found that YUP, the genetic locus responsible for a large portion of floral color variation and previously shown to be important in bee interactions with other Mimulus species, also played an important role in this novel attraction. These results together suggest that the attraction of new pollinators to hybrid plants could be an underexplored avenue for pollinator shift and speciation.

A PRELIMINARY STUDY OF FLORAL DEVELOPMENT AND BREEDING SYSTEM OF Orthosiphon aristatus (BLUME) MIQ

Orthosiphon aristatus (Blume) Miq. belongs to the family Lamiaceae. There are two varieties, white corolla (OAV-1) and purple corolla (OAV-2) varieties. An observation on inflorescence and flower development of O. aristatus was conducted alongside with the study on its breeding system. Inflorescence of OAV-1 and OAV-2 varieties started to develop approximately two and a half months after transplanting the rooted cuttings. The initiation of inflorescence until the senescence took about 75 days. Flower buds started to appear on the inflorescence approximately after 17 days of the commencement of inflorescence development. The development from flower bud until flower senescence took around 50 days. The artificial pollination tests, however, suggested that O. aristatus is predominantly a self-pollinated species.

Spiranthes minamitaniana (Orchidaceae), a new orchid species from Japan based on morphological and phylogenetic data

The delimitation of species within the genus Spiranthes (Orchidaceae) is problematic due to the intricate interplay of phenotypic plasticity, ecological variation, and hybridization, making identification that relies solely on morphology inadequate. In light of morphological and phylogenetic data, we have described a self-pollinating species, S. minamitaniana within the Spiranthes sinensis species complex. Spiranthes minamitaniana shares several morphological features with S. hachijoensis, such as glabrous inflorescence rachis, ovaries, and sepals, a degenerated rostellum, pollinia without a viscidium, and papillate lip basal callosities. However, it can be distinguished by several morphological traits, including semicircular, weakly- or non-3-lobed stigma, and more conspicuous anther cap. Although S. minamitaniana is morphologically most similar to S. hachijoensis, phylogenetic analysis strongly suggested that S. minamitaniana has independently acquired a selfing reproductive mode and constitutes a separate genetic cluster from it. As such, this taxon is morphologically and phylogenetically distinct from other closely related species. Consequently, this taxon is morphologically and phylogenetically distinct from other closely related species.

Chemical emasculation in cowpea (Vigna unguiculata (L.) Walp.) and dicotyledonous model species using trifluoromethanesulfonamide (TFMSA)

Hybridization plays an indispensable role in creating the diversity associated with plant evolution and genetic improvement of crops. Production of hybrids requires control of pollination and avoidance of self-pollination for species that are predominantly autogamous. Hand emasculation, male sterility genes or male gametocides have been used in several plant species to induce pollen sterility. However, in cowpea (Vigna unguiculata (L.) Walp), a self-pollinated cleistogamous dryland crop, only hand emasculation is used, but it is tedious and time-consuming. In this study, male sterility was effectively induced in cowpea and two dicotyledonous model species (Arabidopsis thaliana (L.) Heynh. and Nicotiana benthamiana Domin) using trifluoromethanesulfonamide (TFMSA). Pollen viability assays using Alexander staining showed that 30 ml of 1000 mg/l TFMSA with two-time treatments of one-week interval at the early stage of the reproductive phase under field or greenhouse conditions induced 99% pollen sterility in cowpea. TFMSA treatment induced non-functional pollen in diploid A. thaliana at two-time treatment of 10 ml of 125–250 mg/l per plant and N. benthamiana at two-time treatment of 10 ml of 250–1000 mg/l per plant. TFMSA-treated cowpea plants produced hybrid seeds when used as the female parent in crosses with non-treated plants used as male parents, suggesting that TFMSA had no effect on female functionality in cowpea. The ease of TFMSA treatment and its effectiveness to induce pollen sterility in a wide range of cowpea genotypes, and in the two model plant species tested in this study, may expand the scope of techniques for rapid pollination control in self-pollinated species, with potential applications in plant breeding and plant reproduction science.

Climate Change Impacts on Seed Production and Quality: Current Knowledge, Implications, and Mitigation Strategies

Climate change is real and inevitable, incessantly threatening the terrestrial ecosystem and global food security. Although the impacts of climate change on crop yield and the environment have received much attention in recent years, there are few studies on its implications for the production of high-quality seeds that provide the basic input for food production. Seeds are the primary planting material for crop cultivation and carry most new agricultural technologies to the field. Climatic abnormalities occurring at harvest and during the post-harvest stages may not always severely impact seed yield but can reduce the morphological, physiological and biochemical quality, ultimately reducing the field performance and planting value of the seed lot. In our preliminary data mining that considered the first 30 species appearing in the search results, seed setting, seed yield and seed quality parameters under temperature, CO2 and drought stresses showed differential response patterns depending on the cotyledon number (monocots vs. dicots), breeding system (self- vs. cross-pollinated), life cycle (annual vs. perennial) and maturity time (seed setting in cooler vs. hotter months). The relative proportions of the 30 species showed that germination and seedling vigour are adversely affected more in dicots and self-pollinated annual species that set seeds in hotter months. Together, these impacts can potentially reduce the quantity and quality of seeds produced. Immediate attention and action are required to understand and mitigate the detrimental impacts of climate change on the production and supply of high-quality seeds. This review summarises the current knowledge on this aspect, predicts the future implications and suggests some potential mitigation strategies in the context of projected population growth, climate change and seed requirement at the global level.

Lost in translation: Molecular basis of reduced flower coloration in a self-pollinated monkeyflower (Mimulus) species.

Phenotypic evolution is usually attributed to changes in protein function or gene transcription. In principle, mutations that affect protein abundance through enhancing or attenuating protein translation also could be an important source for phenotypic evolution. However, these types of mutations remain largely unexplored in the studies of phenotypic variation in nature. Through fine-scale genetic mapping and functional interrogation, we identify a single nucleotide substitution in an anthocyanin-activating R2R3-MYB gene causing flower color variation between a pair of closely related monkeyflower (Mimulus) species, the hummingbird-pollinated Mimulus cardinalis, and self-pollinated Mimulus parishii. This causal mutation is located in the 5′ untranslated region and generates an upstream ATG start codon, leading to attenuated protein translation and reduced flower coloration in the self-pollinated species. Together, our results provide empirical support for the role of mutations affecting protein translation, as opposed to protein function or transcript level, in natural phenotypic variation.

Morpho-molecular genetic diversity and population structure analysis in garden pea (Pisum sativum L.) genotypes using simple sequence repeat markers.

Garden pea (Pisum sativum L.) is a self-pollinated plant species which played an important role for the foundation of modern genetics. Genetic diversity among 56 garden pea genotypes was assessed using 12 morphological descriptors, 19 quantitative traits and 8 simple sequence repeat (SSR) markers. Eight morphological descriptors were found polymorphic, and highest Shannon diversity index was recorded for pod curvature (1.18). Mahalanobis D2 illustrating genetic divergence arranged 56 genotypes into six clusters, with the highest inter-cluster distance between clusters IV and VI (18.09). The average values of Na (number of alleles), Ne (effective number of alleles), I (Shannon's Information index), PIC (polymorphism information content), Ho (observed heterozygosity) and He (expected heterozygosity) were 3.13, 1.85, 0.71, 0.36, 0.002 and 0.41, respectively. Pair wise genetic distance among all pairs of the genotypes varied from 0.33 to 1.00 with an average of 0.76. Based on genetic distance, the genotypes were classified into two main clusters (A and B) by cluster analysis, whereas structure analysis divided the genotypes into four sub-populations. The SSR makers indicated that present of genetic variability among the studied genotypes. When, we compared the groups formed by agro-morphological and molecular data, no genotypes were observed, indicating that both stages of characterization are crucial for a better understanding of the genetic variability. Hybridization between genetically diverse genotypes can be exploited to expend the genetic variability and introduce new traits in the pea breeding program.

The pollen dispersal ability for cross-pollination in winter wheat (Triticum aestivum L.) is related to anther extrusion capability rather than to pollen output

IntroductionTriticum aestivum is a self-pollinated species with a high proportion of cleistogamous flowers constituting a blockade for out-crossing. Therefore, male parent characteristics are the main goal in the production of hybrid wheat seeds. MethodsIn a two-year study conducted in Lublin (Poland), the features of flowering biology that can be relevant for the cross-pollination ability of the pollen donor were investigated in winter wheat genotypes. In particular, the anther extrusion capacity, pollen production at the anther and spike levels, and pollen biological value were examined. The pollen production in the anthers was established using an electronic particle counter. The pollen biological value was assessed based on viability (acetocarmine staining) and in vitro pollen germination (raffinose medium) assays. ResultsThe inter-genotype effect for anther extrusion was significantly correlated to the post-anthesis filament length. The anther extrusion for the individual genotypes fluctuated significantly between the years of the study; however, the interaction with meteorological factors was not assessed. The considerable inter-genotype variability in the pollen donor capability for cross-pollination was highly significantly correlated with the proportion of extruded anthers rather than pollen production, indicating that anther extrusion plays a more important role in the contribution to the final amount of pollen available for cross pollination. The pollen viability determined by staining was high (between 97.5% and 100%), while the in vitro pollen germination varied substantially among the genotypes (between 17.3% and 87.8%). ConclusionThe pollen dispersal ability for cross-pollination in wheat can be improved mainly by increasing the proportion of extruded anthers; however, the pollen production and pollen germination potential cannot be ignored if complete characterization of the ideal male idiotype is to be achieved.

Evidence for the Application of Emerging Technologies to Accelerate Crop Improvement - A Collaborative Pipeline to Introgress Herbicide Tolerance Into Chickpea.

Accelerating genetic gain in crop improvement is required to ensure improved yield and yield stability under increasingly challenging climatic conditions. This case study demonstrates the effective confluence of innovative breeding technologies within a collaborative breeding framework to develop and rapidly introgress imidazolinone Group 2 herbicide tolerance into an adapted Australian chickpea genetic background. A well-adapted, high-yielding desi cultivar PBA HatTrick was treated with ethyl methanesulfonate to generate mutations in the ACETOHYDROXYACID SYNTHASE 1 (CaAHAS1) gene. After 2 years of field screening with imidazolinone herbicide across >20 ha and controlled environment progeny screening, two selections were identified which exhibited putative herbicide tolerance. Both selections contained the same single amino acid substitution, from alanine to valine at position 205 (A205V) in the AHAS1 protein, and KASP™ markers were developed to discriminate between tolerant and intolerant genotypes. A pipeline combining conventional crossing and F2 production with accelerated single seed descent from F2:4 and marker-assisted selection at F2 rapidly introgressed the herbicide tolerance trait from one of the mutant selections, D15PAHI002, into PBA Seamer, a desi cultivar adapted to Australian cropping areas. Field evaluation of the derivatives of the D15PAHI002 × PBA Seamer cross was analyzed using a factor analytic mixed model statistical approach designed to accommodate low seed numbers resulting from accelerated single seed descent. To further accelerate trait introgression, field evaluation trials were undertaken concurrent with crop safety testing trials. In 2020, 4 years after the initial cross, an advanced line selection CBA2061, bearing acetohydroxyacid synthase (AHAS) inhibitor tolerance and agronomic and disease resistance traits comparable to parent PBA Seamer, was entered into Australian National Variety Trials as a precursor to cultivar registration. The combination of cross-institutional collaboration and the application of novel pre-breeding platforms and statistical technologies facilitated a 3-year saving compared to a traditional breeding approach. This breeding pipeline can be used as a model to accelerate genetic gain in other self-pollinating species, particularly food legumes.

Transfer of resistance alleles from herbicide-resistant to susceptible grass weeds via pollen-mediated gene flow

AbstractThe objective of this paper was to review the reproductive biology, herbicide-resistant (HR) biotypes, pollen-mediated gene flow (PMGF), and potential for transfer of alleles from HR to herbicide-susceptible grass weeds including barnyardgrass, creeping bentgrass, Italian ryegrass, johnsongrass, rigid (annual) ryegrass, and wild oats. The widespread occurrence of HR grass weeds is at least partly due to PMGF, particularly in obligate outcrossing species such as rigid ryegrass. Creeping bentgrass, a wind-pollinated turfgrass species, can efficiently disseminate herbicide resistance alleles via PMGF and movement of seeds and stolons. The genusAgrostiscontains about 200 species, many of which are sexually compatible and produce naturally occurring hybrids and hybrids with species in the genusPolypogon. The self-incompatibility, extremely high outcrossing rate, and wind pollination in Italian ryegrass clearly point to PMGF as a major mechanism by which herbicide resistance alleles can spread across agricultural landscapes, resulting in abundant genetic variation within populations and low genetic differentiation among populations. Italian ryegrass can readily hybridize with perennial ryegrass and rigid ryegrass due to their similarity in chromosome numbers (2n= 14), resulting in interspecific gene exchange. Johnsongrass, barnyardgrass, and wild oats are self-pollinated species, so the potential for PMGF is relatively low and limited to short distances; however, seeds can easily shatter upon maturity before crop harvest, leading to wider dispersal. The occurrence of PMGF in reviewed grass weed species, even at a low rate, is greater than that of spontaneous mutations conferring herbicide resistance in weeds and thus can contribute to the spread of herbicide resistance alleles. This review indicates that the transfer of herbicide resistance alleles occurs under field conditions at varying levels depending on the grass weed species.

Cytoplasmic and nuclear genome variations of rice hybrids and their parents inform the trajectory and strategy of hybrid rice breeding

The male sterility (MS) line is a prerequisite for efficient production of hybrid seeds in rice, a self-pollinating species. MS line breeding is pivotal for hybrid rice improvement. Understanding the historical breeding trajectory will help to improve hybrid rice breeding strategies. Maternally inherited cytoplasm is an appropriate tool for phylogenetic reconstruction and pedigree tracing in rice hybrids. In this study, we analyzed the cytoplasmic genomes of 1495 elite hybrid rice varieties and identified five major types of cytoplasm, which correspond to different hybrid production systems. As the cytoplasm donors for hybrids, 461 MS lines were also divided into five major types based on cytoplasmic and nuclear genomic architecture. Specific core accessions cooperating with different fertility-associated genes drove the sequence divergence of MS lines. Dozens to hundreds of convergent and divergent selective sweeps spanning several agronomic trait-associated genes were identified among different types of MS lines. We further analyzed the cross patterns between different types of MS lines and their corresponding restorers. This study systematically analyzed the cytoplasmic genomes of rice hybrids revealed their relationships with nuclear genomes of MS lines, and illustrated the trajectory of hybrid rice breeding and the strategies for breeding different types of MS lines providing new insights for future improvement of hybrid rice.

TWAS results are complementary to and less affected by linkage disequilibrium than GWAS

A genome-wide association study (GWAS) is used to identify genetic markers associated with phenotypic variation. In contrast, a transcriptome-wide association study (TWAS) detects associations between gene expression levels and phenotypic variation. It has previously been shown that in the cross-pollinated species, maize (Zea mays), GWAS, and TWAS identify complementary sets of trait-associated genes, many of which exhibit characteristics of true positives. Here, we extend this conclusion to the self-pollinated species, Arabidopsis thaliana and soybean (Glycine max). Linkage disequilibrium (LD) can result in the identification, via GWAS, of false-positive associations. In all three analyzed plant species, most trait-associated genes identified via TWAS are well separated physically from other candidate genes. Hence, TWAS is less affected by LD than is GWAS, demonstrating that TWAS is particularly well suited for association studies in genomes with slow rates of LD decay, such as soybean. TWAS is reasonably robust to the plant organs/tissues used to determine expression levels. In summary, this study confirms that TWAS is a promising approach for accurate gene-level association mapping in plants that is complementary to GWAS, and established that TWAS can exhibit substantial advantages relative to GWAS in species with slow rates of LD decay.

Genetic analysis of the homogeneity of soybean varieties by microsatellite loci

Soybean is the main protein and oil crop in the world. The production of soybeans is increasing every year. The special attention is given to the breeding and seed production of this crop due to its high economic importance. Currently, the morphological characteristics are used to characterize new varieties, applied for a patent. The analysis of polymorphism of the lengths of microsatellite loci (SSR) allows both to identify genotypes exactly and to evaluate genetic homogeneity within a variety. Soybean is a self-pollinating plant species; cross-pollination is extremely rare. The study of the genetic homogeneity of modern varieties of cultivated soybean showed the presence of genetic variability in five of the twenty studied varieties. We observed the heterogeneity of the varieties from one to four DNA SSR loci. We found the maximum level of intravarietal polymorphism in the variety Vilana beta – 46 % by the Soypr1 locus.

Extraction of high-quality genomic DNA and identification of different DNA barcoding markers for chickpea (Cicer arietinum L.)

The genetic studies of individual plants, especially self-pollinated species like chickpea need to be evaluated at the DNA level with the help of molecular markers for identifying genetic variations among the plants. High-quality DNA extraction is a prerequisite for genetic studies. Extraction of intact genomic DNA with high – molecular mass is essential for the study of many molecular biology applications like Polymerase Chain Reaction, endonuclease restriction digestion, southern blot analysis, and also for the construction of a genomic library. Several plant DNA extraction methods are available, even though the DNA isolation methods that give good yield employing both quantity and quality is quite difficult especially for self-pollinated crops like a chickpea. This work was focused on developing a standard protocol for the extraction of genomic DNA and identifying different barcoding markers. The result revealed that the CTAB extraction method with slight modification in protocol had been optimized for DNA isolation. The purified DNA, which was isolated through the CTAB method, had excellent spectral qualities and is efficiently digested by a restriction endonuclease, and is found to be more suitable for long-fragment PCR amplification. DNA barcoding is considered as a promising tool because it provides a practical and standard identification of plants. The isolated DNA sample was processed with a classical DNA barcoding approach by amplifying and sequencing with a universal primer. According to the result, among the different barcoding markers studied, the RbcL and Mat K were found to given the best result for molecular species identification in chickpea.

Food/Feed and Environmental Risk Assessment of Genetically Modified Glufosinate-tolerant Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Food/Feed and Environmental Risk Assessment of Genetically Modified Glufosinate-tolerant Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Integration, abundance, and transmission of mutations and transgenes in a series of CRISPR/Cas9 soybean lines

BackgroundAs with many plant species, current genome editing strategies in soybean are initiated by stably transforming a gene that encodes an engineered nuclease into the genome. Expression of the transgene results in a double-stranded break and repair at the targeted locus, oftentimes resulting in mutation(s) at the intended site. As soybean is a self-pollinating species with 20 chromosome pairs, the transgene(s) in the T0 plant are generally expected to be unlinked to the targeted mutation(s), and the transgene(s)/mutation(s) should independently assort into the T1 generation, resulting in Mendellian combinations of transgene presence/absence and allelic states within the segregating family. This prediction, however, is not always consistent with observed results.ResultsIn this study, we investigated inheritance patterns among three different CRISPR/Cas9 transgenes and their respective induced mutations in segregating soybean families. Next-generation resequencing of four T0 plants and four T1 progeny plants, followed by broader assessments of the segregating families, revealed both expected and unexpected patterns of inheritance among the different lineages. These unexpected patterns included: (1) A family in which T0 transgenes and mutations were not transmitted to progeny; (2) A family with four unlinked transgene insertions, including two respectively located at paralogous CRISPR target break sites; (3) A family in which mutations were observed and transmitted, but without evidence of transgene integration nor transmission.ConclusionsGenome resequencing provides high-resolution of transgene integration structures and gene editing events. Segregation patterns of these events can be complicated by several potential mechanisms. This includes, but is not limited to, plant chimeras, multiple unlinked transgene integrations, editing of intended and paralogous targets, linkage between the transgene integration and target site, and transient expression of the editing reagents without transgene integration into the host genome.

Food, Feed and Environmental Risk Assessment of Glufosinatetolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing Under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Food, Feed and Environmental Risk Assessment of Glufosinatetolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing Under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)