Benchmark study on glyphosate-resistant crop systems in the United States. Part 2: Perspectives

A six-state, 5 year field project was initiated in 2006 to study weed management methods that foster the sustainability of genetically engineered (GE) glyphosate-resistant (GR) crop systems. The benchmark study field-scale experiments were initiated following a survey, conducted in the winter of 2005-2006, of farmer opinions on weed management practices and their views on GR weeds and management tactics. The main survey findings supported the premise that growers were generally less aware of the significance of evolved herbicide resistance and did not have a high recognition of the strong selection pressure from herbicides on the evolution of herbicide-resistant (HR) weeds. The results of the benchmark study survey indicated that there are educational challenges to implement sustainable GR-based crop systems and helped guide the development of the field-scale benchmark study. Paramount is the need to develop consistent and clearly articulated science-based management recommendations that enable farmers to reduce the potential for HR weeds. This paper provides background perspectives about the use of GR crops, the impact of these crops and an overview of different opinions about the use of GR crops on agriculture and society, as well as defining how the benchmark study will address these issues.     

Natural tolerance of Cuscuta campestris to herbicides inhibiting amino acid biosynthesis

The response of Cuscuta campestris Yuncker, a non‐specific above‐ground holoparasite, to amino acid biosynthesis inhibitor (AABI) herbicides, was compared with other resistant and sensitive plants in dose–response assays carried out in Petri dishes. Cuscuta campestris was found to be much more resistant to all AABI herbicides tested. The I₅₀ value of C. campestris growth inhibition by glyphosate was eightfold higher than that of transgenic, glyphosate‐resistant cotton (RR‐cotton). The I₅₀ value for C. campestris shoot growth inhibition by sulfometuron was above 500 μM, whereas that of sorghum roots was only 0.004 μM. Cuscuta campestris exposed to glyphosate gradually accumulated shikimate, confirming herbicide penetration into the parasite and interaction with an active form of the target enzyme of the herbicide, 5‐enolpyruvylshikimate‐3‐phosphate synthase. More than half of the C. campestris plants associated with transgenic, glyphosate‐resistant sugarbeet (RR‐sugarbeet) treated with glyphosate or with transgenic, sulfometuron‐resistant tomato (SuR‐tomato) treated with sulfometuron recovered and resumed regular growth 20–30 days after treatment. New healthy stems developed, followed by normal flowering and seed setting. The results of the current study demonstrate the unique capacity of C. campestris to tolerate high rates of AABI. The mechanism of this phenomenon is yet to be elucidated.     

A survey of Lolium rigidum populations in citrus orchards: Factors explaining infestation levels

The presence of herbicide‐resistant Lolium rigidum in Mediterranean (Spanish) citrus orchards was reported in 2005 and it poses a serious threat to crop management. The main objective of this research was to investigate which components could be responsible for the persistence of annual ryegrass populations in Mediterranean mandarin and orange orchards. This is the first study regarding L. rigidum populations in Mediterranean citrus orchards. Surveys were conducted in 55 commercial citrus orchards in eastern Spain in 2013 by interviewing technicians who were working in cooperatives about crop management. The level of infestation by L. rigidum and the presence of harvester ants (Messor barbarus) then were estimated in the same orchards. The variables were subjected to a two‐dimensional analysis and both univariate and multivariate logistic regression models were fitted for each of the three L. rigidum density levels that had been established. The multivariate models showed the significant factors that were associated with various L. rigidum densities: (i) at a low density, the herbicides that were applied, the number of applications in 2013 and the type of irrigation (flood or drip); (ii) at a medium density, the presence of harvester ants; and (iii) at a high density, the herbicides that were applied in 2013. The results indicated that drip irrigation and one application of glyphosate mixed with other herbicides (or herbicides other than glyphosate) were associated with a lower L. rigidum density. The alternative management options that are presented here should help farmers to reduce weed problems in Mediterranean citrus orchards. Future research is required to better understand the presence of herbicide‐resistant populations, as well as the possible beneficial presence of granivorous ant species.     

Evolution and spread of glyphosate resistance in Conyza bonariensis in California and a comparison with closely related Conyza canadensis

Glyphosate‐resistant weeds are an increasing problem in perennial cropping systems in the Central Valley of California, USA. To elucidate the evolutionary origins and spatial spread of resistance, we investigated the geographical distribution of glyphosate resistance and the population genetic diversity and structure of Conyza bonariensis and compared the results with previously studied C. canadensis. Thirty‐five populations from orchards and vineyards across the Central Valley were sampled. Population genetic structure was assessed using microsatellite markers. Population‐level resistance was assessed in glasshouse screening of plants grown from field‐collected seed. Bayesian clustering and analyses of multilocus genotypes indicated multiple origins of resistance, as observed in C. canadensis. Pairwise F_ST analysis detected spatial spread of resistance in the south of the Central Valley, also similar to C. canadensis. The results strongly indicate that the southern valley was an environment markedly more suitable than the northern valley for resistance spread and that spread in Conyza species was driven by increased uniformity of strong selection in the southern valley, due to recent regulation on herbicides other than glyphosate. Accordingly, resistant C. canadensis individuals occurred at high frequencies only in the southern valley, but interestingly resistant C. bonariensis occurred at high frequencies throughout the valley. Expression of resistance showed varying degrees of plasticity in C. bonariensis. The lower selfing rate and substantially greater genotypic diversity in C. bonariensis, relative to C. canadensis, indicate greater evolutionary potential over shorter time periods. Interspecific hybridisation was detected, but its role in resistance evolution remains unclear.     

分子对接阐明草甘膦与水稻醛酮还原酶(OsALR2)的作用及OsALR2的表达纯化

草甘膦是一种广谱高效的有机磷类除草剂, 因其对水稻等禾本科作物缺乏选择性, 导致其在水稻田中的应用受到限制。杂草中醛酮还原酶(aldo-keto reductase, AKR)具有降解小分子农药草甘膦的功能, 但是水稻响应草甘膦的作用机制未知。本文通过生物信息学分析、分子对接研究草甘膦与其在水稻中的靶标AKR蛋白OsALR2的互作机制。采用TMHMM 2.0和DiANNA 1.1 web server 等在线软件对 OsALR2 蛋白的跨膜特性和二硫键含量等生物学特性进行了分析, 结果表明OsALR2分子量为 35 kD, 等电点为 5.89; OsALR2不是膜蛋白, 但是含有3组二硫键。使用AlphaFold 预测OsALR2 蛋白的三级结构, 并利用Auto Dock Vina将草甘膦与OsALR2进行分子对接, 结果显示, 最低结合能为 -13.4 kcal/mol, 草甘膦与OsALR2的结合主要通过氢键作用。通过Ramachandran图, ERRAT和Verify 3D证明对接获得的三维结构是合理的。进一步通过分子克隆技术构建重组表达载体 pET-32a-OsALR2 并进行原核表达, 结果显示OsALR2主要以包涵体的形式表达, 对包涵体进行变复性后, 蛋白表现出酶活性且蛋白结构有较好的均一性。本文的研究结果助于阐明草甘膦与水稻蛋白互作的分子机制, 为研发抗草甘膦水稻新品种和新型小分子农药提供理论指导。     

Shikimate accumulation in nine weedy species following glyphosate application

Glyphosate is commonly used in a variety of weed control scenarios and plants respond biochemically by continuing to attempt to push carbon through the shikimate pathway. The result is an accumulation of shikimate in sensitive plants in the days immediately after application. This research determined shikimate accumulation dynamics in nine problematic plant species from 0 to 6 days after treatment (DAT). Ambrosia artemisiifolia, Trifolium repens and Conyza canadensis showed rising concentrations through 6 DAT to >4000 mg L^?1. Chenopodium album, Xanthium strumarium and Urochloa platyphylla showed concentrations levelling off to a constant of about 2000 mg L^?1. Ambrosia trifida, Amaranthus palmeri and Polygonum pensylvanicum generally showed lower levels of shikimate accumulation and concentrations were decreasing after 3 DAT. No apparent relationships were evident between shikimate accumulation pattern and plant growth rate or speed of glyphosate activity. There also appeared to be no pattern in the shikimate accumulation trends that would allow for an informed decision as to which of these species would be most likely to develop glyphosate resistance. Shikimate concentrations were in agreement between field and glasshouse conditions for C. canadensis, but did not agree with those from A. palmeri.     

藜、铁苋菜和苘麻对草甘膦的耐受性及其与莽草酸含量的关系

采用整株生物测定法研究了藜、铁苋菜和苘麻对草甘膦的耐受性,药后14 d测定结果表明,草甘膦对上述3种杂草的ED50分别为215.27、954.34、1 522.54 g a.i./hm~2。通过比较杂草植株地上部莽草酸积累量的变化发现,草甘膦1 230 g a.i./hm~2处理后,藜、铁苋菜和苘麻莽草酸积累量最大值分别为1 400.65、1247.19、581.28μg/g,莽草酸积累量越少的杂草对草甘膦耐受性也越强。药后5 d,对杂草不同部位莽草酸积累量的比较显示,3种杂草顶部叶片莽草酸积累量明显大于根部,敏感种和耐受种相比,顶部叶片莽草酸积累量的差异更为明显,该部位可以准确地评价杂草对草甘膦的耐受程度。     

甲磺隆和草甘膦对空心莲子草乙酰乳酸合酶活性和莽草酸含量的影响

为研究甲磺隆和草甘膦对空心莲子草的作用机理,探索其最佳的使用技术,采用有效成分为30 g/hm2和60 g/hm2 的甲磺隆处理空心莲子草,能明显抑制其茎和根乙酰乳酸合酶的比活性;有效成分为1537.5g/ hm2和3075g/ hm2 的草甘膦处理则能明显抑制空心莲子草莽草酸含量的积累;施用甲磺隆和草甘膦对空心莲子草生长的抑制作用随药剂浓度的提高而增大.二次施用草甘膦(间隔30天)的结果表明,低剂量处理对抑制空心莲子草根组织莽草酸含量的积累更明显.