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In addition, we discuss the progress that has been made towards engineering salt tolerance in crops, including marker-assisted selection and gene stacking techniques.
Several genes, including Atriplex hortensis betaine aldehyde dehydrogenase (badh) involved in the accumulation of glycine betaine giving tolerance to salt stress [24], isoflavone synthase (ifs) from Medicago truncatula that a key enzyme in the flavonoids/isoflavonoids pathway [25], a phosphinothricin resistance gene (bar) and two reporter genes, gus and gfp, were chosen for gene stacking.
This modified method for marker-free site-specific integration is suitable for iterative gene stacking.
CG and nematode resistance gene stacking controlled CG and RLNs simultaneously in walnuts.
This process could be repeated to achieve gene stacking into the selected chromosomal sites.
Gene stacking methods that do not involve the use of nucleases have been reported.
Our study shows the promise of the gene stacking approach for engineering plants with improved properties for biofuel applications.
Gene stacking into pre-determined genomic sites depends on mechanisms of targeted DNA integration and recycling of selectable marker genes.
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Against this, the fact that most known Z. tritici isolates are virulent to most Stb genes (Section 5.1) suggests that the resistance achieved by gene-stacking may not be durable.
The gene-stacking approach entails manipulation of two or more desirable enzymes mediating the ROS turnover and scavenging pathways, in improving the abiotic stress tolerance in plants.
The relevance and potential of these promoters in biotechnology has been documented [ 4- 6], particularly for use in gene-stacking approaches where more than one gene is required to confer a particular trait trangenically or more than one trait is being conferred e.g. resistance to a suite of pests [ 7].
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