Dataset Information

Development of non-transgenic glyphosate tolerant wheat by TILLING.

ABSTRACT: Glyphosate (N-phosphonomethyl-glycine) is the world's most widely used broad spectrum, post-emergence herbicide. It inhibits the chloroplast-targeted enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19), a component of the plant and microorganism-specific shikimate pathway and a key catalyst in the production of aromatic amino acids. Variants of EPSPS that are not inhibited by glyphosate due to particular amino acid alterations in the active site of the enzyme are known. Some of these variants have been identified in weed species that have developed resistance to glyphosate because of the strong selective pressure of continuous, heavy glyphosate use. We have used TILLING (Targeting Induced Local Lesions in Genomes), a non-transgenic, target-selected, reverse genetics, mutation breeding technique, and conventional genetic crosses, to identify and combine, through two rounds of mutagenesis, wheat lines having both T102I and P106S (so-called TIPS enzyme) mutations in both the A and the D sub-genome homoeologous copies of the wheat EPSPS gene. The combined effects of the T102I and P106S mutations are known from previous work in multiple species to minimize the binding of the herbicide while maintaining the affinity of the catalytic site for its native substrates. These novel wheat lines exhibit substantial tolerance to commercially relevant levels of glyphosate.

SUBMITTER: Moehs CP

PROVIDER: S-EPMC8443057 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Development of non-transgenic glyphosate tolerant wheat by TILLING.

Moehs Charles P CP Austill William J WJ Facciotti Daniel D Holm Aaron A Loeffler Dayna D Lu Zhongjin Z Mullenberg Jessica C JC Slade Ann J AJ Steine Michael N MN van Boxtel Jos J McGuire Cate C

PloS one 20210915 9

Glyphosate (N-phosphonomethyl-glycine) is the world's most widely used broad spectrum, post-emergence herbicide. It inhibits the chloroplast-targeted enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19), a component of the plant and microorganism-specific shikimate pathway and a key catalyst in the production of aromatic amino acids. Variants of EPSPS that are not inhibited by glyphosate due to particular amino acid alterations in the active site of the enzyme are known. Some ...[more]

PMID: 34525118

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Project description:Mutagenesis is an important tool in crop improvement. However, the hexaploid genome of wheat (Triticum aestivum L.) presents problems in identifying desirable genetic changes based on phenotypic screening due to gene redundancy. TILLING (Targeting Induced Local Lesions IN Genomes), a powerful reverse genetic strategy that allows the detection of induced point mutations in individuals of the mutagenized populations, can address the major challenge of linking sequence information to the biological function of genes and can also identify novel variation for crop breeding. Wheat is especially well-suited for TILLING due to the high mutation densities tolerated by polyploids. However, only a few wheat TILLING populations are currently available in the world, which is far from satisfying the requirement of researchers and breeders in different growing environments. In addition, current TILLING screening protocols require costly fluorescence detection systems, limiting their use, especially in developing countries. We developed a new TILLING resource comprising 2610 M(2) mutants in a common wheat cultivar 'Jinmai 47'. Numerous phenotypes with altered morphological and agronomic traits were observed from the M(2) and M(3) lines in the field. To simplify the procedure and decrease costs, we use unlabeled primers and either non-denaturing polyacrylamide gels or agarose gels for mutation detection. The value of this new resource was tested using PCR with RAPD and Intron-spliced junction (ISJ) primers, and also TILLING in three selected candidate genes, in 300 and 512 mutant lines, revealing high mutation densities of 1/34 kb by RAPD/ISJ analysis and 1/47 kb by TILLING. In total, 31 novel alleles were identified in the 3 targeted genes and confirmed by sequencing. The results indicate that this mutant population represents a useful resource for the wheat research community. We hope that the use of this reverse genetics resource will provide novel allelic diversity for wheat improvement and functional genomics.

Project description:Conventional farming poses threats to sustainable agriculture in growing food demands and increasing flooding risks. This research introduces a Bayesian Belief Network (BBN) to address these concerns. The model explores tillage adaptation for flood management in soils with varying organic carbon (OC) contents for winter wheat production. Three real soils, emphasizing texture and soil water properties, were sourced from the NETMAP soilscape of the Pang catchment area in Berkshire, United Kingdom. Modified with OC content at four levels (1, 3, 5, 7%), they were modeled alongside relevant variables in a BBN. The Decision Support System for Agrotechnology Transfer (DSSAT) simulated datasets across 48 cropping seasons to parameterize the BBN. The study compared tillage effects on wheat yield, surface runoff, and GHG-CO2 emissions, categorizing model parameters (from lower to higher bands) based on statistical data distribution. Results revealed that NT outperformed CT in the highest parametric category, comparing probabilistic estimates with reduced GHG-CO2 emissions from "7.34 to 7.31%" and cumulative runoff from "8.52 to 8.50%," while yield increased from "7.46 to 7.56%." Conversely, CT exhibited increased emissions from "7.34 to 7.36%" and cumulative runoff from "8.52 to 8.55%," along with reduced yield from "7.46 to 7.35%." The BBN model effectively captured uncertainties, offering posterior probability distributions reflecting conditional relationships across variables and offered decision choice for NT favoring soil carbon stocks in winter wheat (highest among soils "NT.OC-7%PDPG8," e.g., 286,634 kg/ha) over CT (lowest in "CT.OC-3.9%PDPG8," e.g., 5,894 kg/ha). On average, NT released minimum GHG- CO2 emissions to "3,985 kgCO2eqv/ha," while CT emitted "7,415 kgCO2eqv/ha." Conversely, NT emitted "8,747 kgCO2eqv/ha" for maximum emissions, while CT emitted "15,356 kgCO2eqv/ha." NT resulted in lower surface runoff against CT in all soils and limits runoff generations naturally for flood alleviation with the potential for customized improvement. The study recommends the model for extensive assessments of various spatiotemporal conditions. The research findings align with sustainable development goals, e.g., SDG12 and SDG13 for responsible production and climate actions, respectively, as defined by the Agriculture and Food Organization of the United Nations.

Dataset Information

Development of non-transgenic glyphosate tolerant wheat by TILLING.

Publications

Development of non-transgenic glyphosate tolerant wheat by TILLING.

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