Development of marker-free transgenic plants is a technical alternative for avoiding concerns about the safety of selectable marker genes used in genetically modified (GM) crops. Here, we describe the construction of a spontaneous self-excision binary vector using an oxidative stress-inducible modified FLP/FRT system and its successful application to produce marker-free transgenic rice plants with enhanced seed tocopherol content. To generate selectable marker-free transgenic rice plants, we constructed a binary vector using the hpt selectable marker gene and the rice codon-optimized FLP (mFLP) gene under the control of an oxidative stress-inducible promoter between two FRT sites, along with multiple cloning sites for convenient cloning of genes of interest. Using this pCMF binary vector with the NtTC gene, marker-free T1 transgenic rice plants expressing NtTC were produced by Agrobacterium-mediated stable transformation using hygromycin as a selective agent, followed by segregation of selectable marker genes. Furthermore, α-, γ-, and total tocopherol levels were significantly increased in seeds of the marker-free transgenic TC line compared with those of wild-type plants. Thus, this spontaneous auto-excision system, incorporating an oxidative stress-inducible mFLP/FRT system to eliminate the selectable marker gene, can be easily adopted and used to efficiently generate marker-free transgenic rice plants. Moreover, nutritional enhancement of rice seeds through elevation of tocopherol content coupled with this marker-free strategy may improve human health and public acceptance of GM rice.
Pepper (Capsicum spp.; Family: Solanaceae; 2n = 24) is an important crop cultivated worldwide for the consumption of its fresh and dried processed fruits. Pepper fruits are used as raw materials in a wide variety of industrial processes. As a multipurpose vegetable crop, there is a need to increase the yield. However, yield productivity of pepper is severely constrained by infectious plant pathogens, including viruses, bacteria, fungi, and oomycetes. The pepper mild mottle virus (PMMoV) is currently one of the most damaging pathogens associated with yield losses in pepper production worldwide. In addition to impacts on pepper productivity, PMMoV has been detected in domestic and aquatic water resources, as well as in the excreta of animals, including humans. Therefore, PMMoV has been suggested as a potential indicator of domestic water quality. These findings present additional concerns and trigger the need to control the infectious pathogen in crop production. This review provides an overview of the distribution, economic impacts, management, and genome sequence variation of some isolates of PMMoV. We also describe genetic resources available for crop breeding against PMMoV.
Nutritional composition is important for assessing the safety of genetically modified (GM) crops for human consumption. Three beta-carotene-enhanced soybean lines were developed by introducing the β-conglycinin promoter::Phytoene synthase-2A-Carotene desaturase/t35S gene cassette into the genome of the commercial Kwangan (Glycine max L.) soybean variety. Transgenic soybeans were successfully detected on beta-carotene productions ranged from 170.47 to 213.58 µg/g. Comparative assessments of nutrition were conducted with 3 transgenic soybeans, their non-GM counterpart, and several commercial soybean varieties. Results indicated that most levels of proximate, fatty acids, amino acids, and vitamins showed non-significant differences between transgenic soybeans and their counterpart, and fit within the reference ranges established for other commercial soybeans and Organization for Economic Cooperation and Development Guidelines. However, significant differences on levels of crude fat, carbohydrate, δ-tocopherol, and oleic acid of transgenic soybeans comparing to those of non-transgenic counterpart Kwangan cannot eliminate the influences of transgene insertion. Alternations on compositions should be definite by further studies, such as transcriptome and metabolome profiling.
14 June 2014 in Plant Cell, Tissue and Organ Culture (PCTOC)
Lettuce (Lactuca sativa) is a globally significant leafy vegetable, valued for both its economic and nutritional contributions. The efficient conservation and use of the lettuce germplasm are crucial for breeding and genetic improvement. This study examined the genetic diversity and population structure of a core collection of the lettuce germplasm using genotyping by sequencing (GBS). A total of 7136 high-quality single-nucleotide polymorphisms (SNPs) were identified across nine chromosomes. Population analysis through Bayesian clustering and discriminant analysis of principal components (DAPC) revealed three distinct genetic clusters. Cluster 2 exhibited the greatest genetic diversity (He = 0.29, I = 0.44), while Cluster 3 had high levels of inbreeding (F = 0.79). Agro-morphological trait evaluation further identified significant differences in leaf length, plant weight, and head height across clusters. These findings provide valuable insights into the genetic and phenotypic diversity of lettuce, facilitating the development of more robust breeding programs. Additionally, the core collection established in this study offers a representative subset of the lettuce germplasm for future genomic research and conservation efforts.