Immunoassays' a sensitive technique has a wide application for detection of antigen, especially for the detection of growing genetically modified crops, which are underwent field trial. Here, we have developed for the quantitative detection of Cry2Ab protein expressed in GM crops. A Cry2Ab-rabbit-IgG act as capture antibody and Cry2Ab mouse monoclonal antibody behave as a detecting antibody, which is employed in sandwich ELISA. A 2% polyvinylpyrrollidone and 1% dithiothritol was utilized in protein extraction from cotton seed to avert interferring agents. The developed assay was validated with GM cotton samples, limit of detection was found 1 pg/ml and range to a limit of quantification 16 pg/ml. The developed immunoassay does not show any cross reactivity with other non target GM proteins like Cry1Ab, Cry1Ac, EPSPS, Vip.

Site-specific and adaptable DNA binding domains are essential modules to develop genome engineering technologies for crop improvement. Transcription activator-like effectors (TALEs) proteins are used to provide a highly specific and adaptable DNA binding modules. TALE chimeric nucleases (TALENs) were used to generate site-specific double strand breaks (DSBs) in vitro and in yeast, Caenorhabditis elegans, mammalian and plant cells. The genomic DSBs can be generated at predefined and user-selected loci and repaired by either the non-homologous end joining (NHEJ) or homology dependent repair (HDR). Thus, TALENs can be used to achieve site-specific gene addition, stacking, deletion or inactivation. TALE-based genome engineering tools should be powerful to develop new agricultural biotechnology approaches for crop improvement. Here, we discuss the recent research and the potential applications of TALENs to accelerate the generation of genomic variants through targeted mutagenesis and to produce a non-transgenic GM crops with the desired phenotype.

This work was carried out in order to develop early flowering barley lines. These lines will be useful to producers by enabling multiple crops within a single season and increasing production. Transgenic barley plants containing the natural early flowering time AtCRY2 allele from the Cape Verde Island (Cvi) ecotype of Arabidopsis have been generated using biolistic transformation. Immature embryo derived calli of two commercially important barley cultivars (El-Dwaser and El-Taif), were transformed using a pCAMBIA-2300 plasmid harboring a genomic fragment containing the AtCRY2-Cvi allele. Transformation was performed utilizing 600 immature embryos for each cultivar. Stable transformation was confirmed in T0 and T1 plants by using genomic PCR, RT-PCR and western blot analysis with AtCRY2 specific primers and antibodies, respectively. The transformation efficiency was 5.6% and 3.4% for El-Dwaser and El-Taif cultivars, respectively. Seeds from several T1 lines were germinated on kanamycin plates and the lines that contained a single locus were selected for further evaluation. The transformed barley plants showed the specific AtCRY2-Cvi flowering phenotype, i.e. early flowering and day length insensitivity, compared to the non transgenic plants. The time to flowering in transgenic T1 plants was assessed and two lines exhibited flowering more than 25 days earlier than the parental cultivars under short day conditions.

Ectoine is a common compatible solute in halophilic bacteria. Its biosynthesis originates from L-aspartate β-semialdehyde and requires three enzymes: L-2, 4-diaminobutyric acid aminotransferase (gene: ect B), L-2,4-diaminobutyric acid acetyl transferase (gene: ect A) and L-ectoine synthase (gene: ect C). Genetically engineered tomato plants expressing the three H. elongata genes (ectA, ectB, and ectC) generated showed no phenotypic abnormality. Expression of the ectoine biosynthetic genes was detected in the T3 transgenic plants by Northern blot analysis. The ectoine accumulating T3 plants were evaluated for salt tolerance by examining their photosynthestic activity, osmotic adjustment and carbon partitioning. Nuclear magnetic resonance (NMR) detected the accumulation of ectoine. The concentration of ectoine increased with increasing salinity. The transgenic lines showed higher activities of peroxidase, while the malondialdehyde (MDA) concentration was decreased under salinity stress condition. In addition, preservation of higher rates of photosynthesis and turgor values as compared to control was evident. Within a week of 13CO2 feeding, salt application led to increases in the partitioning of 13C into roots at the expense of 13C in the other plant parts. These results suggest that under saline conditions ectoine synthesis is promoted in the roots of transgenic plants, leading to an acceleration of sink activity for photosynthate in the roots. Subsequently, root function such as water uptake is improved, compared with wild-type plants. In this way, the photosynthetic rate is increased through enhancement of cell membrane stability in oxidative conditions under salt stress.

As part of the regulatory safety assessment for transgenic crops, the DNA of the genetic insert and insert/flanking-region junctions are searched for random reading frames that could encode unsafe proteins. In addition, the endogenous flanking regions are examined to determine if the insert disrupts endogenous genes or regulatory elements. We discuss the likelihood of these events in the context of traditional breeding and in the context of the probability that cryptic reading frames might encode unsafe proteins. We conclude that there is a lack of theoretical and empirical evidence to suggest that these safety risks are significant or different compared with those of traditional crop breeding.

The accumulation of compatible solutes during stress in plant cell is well documented. Proline is one of these solutes which accumulate in the cytosol in response to drought or salinity stress in plants. Proline has several functions during stress just like osmotic adjustment, osmoprotection, free radical scavenger and antioxidant. Ornithine δ-aminotransferase (δ-OAT) is an important enzyme in proline biosynthetic pathway. It catalyzes the transamination of ornithine to pyrroline-5-carboxylate which can be reduced into proline. Expression of ornithine δ-aminotransferase gene isolated from Vicia villosa (VvOAT) showed protein with a molecular mass of 63 KDa which is compatible with the predicted mass and after VvOAT gene delivery into E. coli host HB101, VvOAT gene enhanced its salt tolerance. Homology modeling of VvOAT was performed based on the crystal structure of the ornithine δ-aminotransferase from humans (PDB code 2OATA). With this model, a flexible docking study with the substrate and inhibitors was performed. The results indicated that PHE170 and ASN171 in VvOAT are the important determinant residues in binding as they have strong hydrogen bonding contacts with the substrate and inhibitors. All the obtained results indicated the efficiency of utilizing this gene in conferring salt tolerance.

Genetically modified organisms are expected to have a large impact on the ability of humanity to feed, fuel and heal itself in light of the growing global change, and adverse geo-climatic conditions anticipated as a result of climate change.

This specially curated group of 178 titles covers subjects ranging from biodiversity conservation to agriculture and ecology to zoology. Books explore urban ecology, climate change, fire mitigation, and more. These titles are included in the CSIRO Publishing BioSelect Collection.