Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.).

Eleusine indica (goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report a successful Agrobacterium-mediated transformation method for goosegrass. Firstly, we optimized conditions for breaking seed dormancy and increasing seed germination rate. A higher callus induction rate from germinated seeds was obtained in N6 than in MS or B5 medium. Then the optimal transformation efficiency of the gus reporter gene was obtained by infection with Agrobacterium tumefaciens culture of OD600 = 0.5 for 30 min, followed by 3 days of co-cultivation with 300 µmol/L acetosyringone. Concentrations of 20 mg L-1 kanamycin and 100 mg L-1 timentin were used to select the transformed calli. The optimal rate of regeneration of the calli was generated by using 0.50 mg L-1 6-BA and 0.50 mg L-1 KT in the culture medium. Then, using this transformation method, we overexpressed the paraquat-resistant EiKCS gene into a paraquat-susceptible goosegrass biotype MZ04 and confirmed the stable inheritance of paraquat-resistance in the transgenic goosegrass lines. This approach may provide a potential mechanism for the evolution of paraquat-resistant goosegrass and a promising gene for the manipulation of paraquat-resistance plants. This study is novel and valuable in future research using similar methods for herbicide resistance.

Overexpression of EiKCS confers paraquat-resistance in rice (Oryza sativa L.) by promoting the polyamine pathway.

BACKGROUND: Paraquat is used widely as one of the bipyridine herbicides, which generates reactive oxygen species to cause cell death. With a growing number of paraquat-resistant weeds, the mechanism of paraquat-resistance in plants remains unclear. This research verified the functions of a previously confirmed putative paraquat-resistant gene, EiKCS, from paraquat-resistant goosegrass by genetic engineering in a single overexpressing line in rice. RESULTS: Overexpression of EiKCS improved paraquat resistance in transgenic rice (KCSox). Pre-applied (12 h) exogenous spermidine (1.5 mmol L-1 ), alleviated the injury of paraquat in rice. Paraquat induced injury in KCSox was 19.57%, which was lower than 32.22% injury it induced in wild-type (WT) rice. The paraquat-resistant mechanism was through the increased activity of antioxidant enzymes and the overproduction of endogenous polyamines. The spermine content in KCSox was more than 30 µg mL-1 , while that in WT rice was less than 5 µg mL-1 . Quantitative proteomics showed that ß-ketoacyl-coenzyme A (CoA) synthase (51.81 folds) encoded by the transgenic EiKCS gene promoted the synthesis of the proteins involved with the polyamine pathway. The synthesized putrescine was promoted by the arginine decarboxylase (ADC) pathway. The spermidine synthase I (1.10-fold) and three eceriferum cofactors (CERs) were responsive to the paraquat stress. We validated putrescine (C18 H20 N2 O2 ) spermidine (C28 H31 N3 O3 ), and spermine (C38 H42 N4 O4 ) in this study. CONCLUSION: EiKCS encoding ß-ketoacyl-CoA synthase from goosegrass has been shown as an ideal candidate gene for engineering genetically modified organism (GMO) crops, as its overexpression does not only bring paraquat-resistance, but also have potential benefits without decreasing yield and rice grain quality. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The Mitochondrial Genome of Eleusine indica and Characterization of Gene Content within Poaceae.

Plant mitochondrial (mt) genome assembly provides baseline data on size, structure, and gene content, but resolving the sequence of these large and complex organelle genomes remains challenging due to fragmentation, frequent recombination, and transfers of DNA from neighboring plastids. The mt genome for Eleusine indica (Poaceae: goosegrass) is comprehensibly analyzed here, providing key reference data for an economically significant invasive species that is also the maternal parent of the allotetraploid crop Finger millet (Eleusine coracana). The assembled E. indica genome contains 33 protein coding genes, 6 rRNA subunits, 24 tRNA, 8 large repetitive regions 15 kb of transposable elements across a total of 520,691 bp. Evidence of RNA editing and loss of rpl2, rpl5, rps14, rps11, sdh4, and sdh3 genes is evaluated in the context of an updated survey of mt genomic gene content across the grasses through an analysis of publicly available data. Hypothesized patterns of Poaceae mt gene loss are examined in a phylogenetic context to clarify timing, showing that rpl2 was transferred to the nucleus from the mitochondrion prior to the origin of the PACMAD clade.

Methiozolin sorption and mobility in sand-based root zones.

BACKGROUND: Methiozolin is a herbicide currently used for annual bluegrass control in golf course putting greens. Previous research indicates that maximum weed control efficacy requires root exposure; however, soil sorption and mobility of methiozolin have not been established. Research was conducted to investigate soil sorption and subsequent desorption by dilution of methiozolin, as well as soil mobility using batch equilibrium experiments and thin-layer chromatography in nine root zones. Evaluations focused on sand-based systems typical of many golf course putting greens. RESULTS: Sorption coefficients (Kd values) ranged from 0.4 to 29.4 mL g(-1) and averaged 13.8 mL g(-1) . Sorption was most influenced by organic matter content; conversely, soil pH had a negligible effect. Methiozolin desorption did not occur with a 0.01 M CaCl2 dilution. Methiozolin mobility was low; retardation factors (Rf values) were <0.05 for all media with ≥0.3% organic matter. Sand (0.1% organic matter) resulted in an Rf value of 0.46. CONCLUSION: Approximately 24% of applied methiozolin is available for root uptake, and mobility is limited, suggesting resistance to loss through leaching displacement.