Creating a novel herbicide-tolerance OsALS allele using CRISPR/Cas9-mediated gene editing

Weeds and weedy rice plague commercial rice fields in many countries. Developingherbicide-tolerance rice is the most efficient strategy to control weed proliferation. CRISPR/Cas9-mediated gene editing, which generates small InDels and nucleotide substitutions atand around target sites using error-prone non-homologous end joining DNA repairing, hasbeen widely adopted for generation of novel crop germplasm with a wide range of geneticvariation in important agronomic traits. We created a novel herbicide-tolerance allele inrice by targeting the acetolactate synthase (OsALS) gene using CRISPR/Cas9-mediated geneediting. The novel allele (G628W) arose from a G-to-T transversion at position 1882 of OsALSand conferred a high level of herbicide tolerance. Transgene-free progeny carryinghomozygous G628W allele were identified and showed agronomic performance similar tothat of wild-type plants, suggesting that the G628W allele is a valuable resource fordeveloping elite rice varieties with strong herbicide tolerance. To promote use of the G628Wallele and to accelerate introgression and/or pyramiding of the G628W allele with other elitealleles, we developed a DNA marker for the G628W allele that accurately and robustlydistinguished homozygous from heterozygous segregants. Our result further demonstratesthe feasibility of CRISPR/Cas9-mediated gene editing in creating novel genetic variation forcrop breeding.

Indoor Propagation of Transgenic Glyphosate-tolerant Rapeseed (Brassica napus L.) and Transfer Breeding for Herbicide Tolerance

[Objectives] This study was conducted to investigate the possibility of screening for herbicide resistance in the early stage of rapes growth. [Methods] Eight treatments were set for a herbicide concentration experiment. [Results] Rape sprouts were highly sensitive to the herbicide glyphosate, and even a very low concentration of glyphosate solution(187 mg/L) was sufficient to kill most of the rape seedlings, while the genetically modified herbicide-tolerant rape could tolerate higher concentrations of the herbicide. Low concentrations of glyphosate could be used for screening at the sprout stage, and the results of herbicide-tolerance screening were consistent with PCR testing. [Conclusions] This study lays a technical and material foundation for accelerating the cultivation of new herbicide-resistant rape varieties suitable for rapeseed production areas in the lower reaches of the Yangtze River.

Glyphosate-Residues in Roundup-Ready Soybean Impair Daphnia magna Life-Cycle

Herbicide tolerant plants such as Roundup-Ready soybean contain residues of glyphosate herbicide. These residues are considered safe and previous animal-feeding-studies have failed to find negative effects related to such chemical residues. The present study tests 8 experimental soy- meal diets as feed in groups (each containing 20 individuals) of test-animals (D. magna). The diets have different levels of glyphosate residues and we show that animal growth, reproductive maturity and number of offspring are correlated with these chemicals. The tested soybeans are from ordinary agriculture in Iowa USA and the residues are below the regulatory limits. Despite this, clear negative effects are seen in life-long feeding. The work enhances the need for including analysis of herbicide residues in future assessment of GMO.