Is dose‐splitting of graminicides as effective as a single application?

Dose‐splitting or reduced dose repeat application, i.e. the application of a herbicide twice to the same plant, is likely to become a more common scenario in future, if farmers adopt the use of novel techniques for prediction of herbicide efficacy shortly after application. Fourteen pot experiments were conducted to study whether graminicides applied to annual grass weeds as dose‐splitting treatments were as effective as single applications. The influence of time interval between applications and the ratio of the doses of the two applications were studied. Two statistical approaches were applied: comparison of the ED₉₀ of the single treatments and a joint action model. The study revealed that flupyrsulfuron, iodosulfuron, sulfosulfuron, clodinafop and glyphosate could be applied as repeat treatments with up to 14 days’ interval between the two applications without loss of activity. In some experiments, particularly with iodosulfuron, dose‐splitting enhanced herbicide performance. By contrast, prosulfocarb responded adversely, most likely due to the pronounced impact of weed growth stage on the activity of this herbicide. The ratio of the doses and the timing between the two applications had no significant influence on herbicide responses. The results of the present studies showed that even if a reduced dose of a graminicide results in an unsatisfactory control and it is necessary to re‐spray, the overall use of herbicide will not increase, if methods are available that can predict the efficacy of a herbicide application within the first 1–2 weeks after application.     

马尾松防火道及果园化学除草试验初报

在马尾松的防火道上用5％森泰颗粒剂和25％森泰水剂、在桃树果园用65％草甘膦和41％春多多进行除草试验。结果表明：马尾松防火道用18．75kg／hm^2的森泰颗粒剂处理效果较好；桃树果园则以1500g／hm^2的草甘膦处理效果最好。     

Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate

Glyphosate-tolerant soybean is the most widely planted genetically modified crop worldwide. However, soybean remains recalcitrant to routine transformation because of the low infection efficiency of Agrobacterium to soybean and lack of useful selectable markers. In this study, several Agrobacterium strains and cell densities were compared by transient expression of the GUS gene. The results showed that Agrobacterium strain Ag10 at cell densities of OD_(600) of 0.6–0.9 yielded the highest infection efficiency in Agrobacterium-mediated soybean cotyledonary node transformation system. Meanwhile, a simple and rapid method was developed for identification of glyphosate tolerance in putative T_0 transgenic plants, consisting of spotting plantlets with 1 μL Roundup~?. The whole cycle of genetic transformation could be shortened to about 3 mon by highly efficient selection with glyphosate during the transformation process and application of the spot assay in putative T_0 transgenic plantlets. The transformation frequency ranged from 2.9 to 5.6%. This study provides an improved protocol for development and identification of glyphosate-tolerant transgenic soybeans.     

Effects of Glyphosate on Soil Salinization and Alkalization in Cotton Fields

We analyzed the effects of glyphosate to the index of salinization and alkalization by simulation in soils collected from cotton fields in Anyang, Henan and Anqing, Anhui, China, in 2014. The results showed that application of glyphosate changed electrical conductivity (EC), total alkalinity (TAL), exchangeable sodium percentage (ESP), sodium adsorption ratio (SAR) and pH of soil in both regions,compared with the blank control. With increased frequency of glyphosate application and increased concentration applied, all indices increased in the Anqing cotton fields soil which were treated with concentrations of 2.5-20 g·L^－1 . However, EC, TAL and ESP increased dynamically in Anyang soil to which high concentrations of 20 g·L^－1 were applied only four consecutive times. After different glyphosate treatments, the soil pH value showed an upward trend but smaller changes. Therefore, after repeated use of glyphosate to soils of two different region soils, although the changes of all indices were within the range of non-saline and non-sodic soils, the soils affected by salt in the glyphosate aqueous solution tended towards saline-sodic soils.     

Biological evidence for a 1:1 Ca2+:glyphosate association in deposit residuals on the leaf surface of barley

It has long been known that calcium ion antagonizes glyphosate, but it was not clear whether the stoichiometry of their interaction is 1:1 or 1:2. Two independent methods were used to determine which stoichiometry was the most probable. First, dose–response curves of barley ( Hordeum vulgare L.) plants treated with glyphosate were determined in the presence of 0, 1.25, 2.5, 5 and 10 mM CaCl₂. The doses of 'free' glyphosate (=not inactivated by calcium ion) were computed using the assumptions of 1:1 and 1:2 stoechiometries. The response curves were redrawn as a function of 'free' glyphosate. Analysis showed that the 1:2 hypothesis could be rejected, whereas the 1:1 hypothesis was highly probable. Second, kinetics of glyphosate penetration into barley leaves were determined in the presence of 0, 2.5 and 10 mM CaCl₂. The concentrations of 'free' glyphosate were computed as above. The kinetics of glyphosate penetration at these concentrations were established and were compared to the kinetics of glyphosate penetration in the presence of CaCl₂. Again, the 1:2 hypothesis was rejected, whereas the 1:1 hypothesis was more probable. These results strongly suggest that the stoichiometry of the Ca²⁺:glyphosate association is 1:1 in deposit residuals on the leaf surface.     

草甘膦抗性菌株的分离鉴定及其抗性基因的克隆

从草甘膦污染土壤中分离获得一株耐受400 mmol/L草甘膦的菌株S1536,该菌株在草甘膦浓度为100～400 mmol/L之间生长迅速,最适pH为6.0,最适生长温度为30℃,具有氨苄青霉素抗性。以S1536基因组DNA为模板,通过通用引物进行扩增、测序获得其16S rDNA序列,在GenBank登录号为MG519831,在NCBI中经BLAST比对与泛菌属（Pantoea sp.）同源性达到99%。用ClustalW对S1536与泛菌属各个种模式菌的 16S rDNA进行多重序列对比,该菌株与Pantoea rodasii同源性最为接近,达到99.2%,故将菌株S1536命名为Pantoea rodasii S1536。该菌株具有较高草甘膦抗性,对温度和pH适应性强,是优良的耐草甘膦菌株,通过全基因组测序及基因注释,获得抗除草剂基因aroA序列,为下一步明确基因功能,挖掘Pantoea rodasii S1536高抗草甘膦的分子机制奠定基础。     

Morphological disparities in the epidermal and anatomical features of the leaf among wild Brassica juncea populations

In the present article, we investigated the epidermal and anatomical features of 10 wild Brassica juncea populations with different susceptibilities to glyphosate in detail. The characteristics of the leaf surface and anatomy could affect the efficacy of glyphosate uptake in weeds through the influence of its adherence and penetration. Our results showed that herbicide-resistant wild populations generally had greater leaf thickness, adaxial cutin thickness, and trichome density in the upper epidermis, but lower spongy tissue thickness and vascular bundle numbers, as compared with the susceptible populations, suggesting that differences in morphological traits among populations might contribute to their variable response to glyphosate. The possible morphological involvement in the differential susceptibility of wild B. juncea populations to glyphosate also was discussed.     

施用草甘膦除草剂对土壤质量影响的研究进展

草甘膦除草剂是农业生产中不可或缺的物资,过量使用会导致大量草甘膦及其衍生物在土壤中残留,对生态环境安全构成威胁。通过系统阐述草甘膦在环境中的迁移转化过程,以及草甘膦对土壤生物及环境质量的影响,着重分析了草甘膦对土壤蚯蚓、微生物群落的毒理效应,草甘膦与土壤其他污染物的协同效应和草甘膦驱动土壤碳氮磷生物地球循环变化。同时指出当前研究的不足,并提出下一步研究的重点:(1)加强草甘膦的降解机制和影响因素的研究,并筛选更高效降解能力的微生物菌株,提高草甘膦降解效率和修复草甘膦污染环境的能力;(2)定期开展农业环境中草甘膦的检测和风险评估,摸清草甘膦在喀斯特区的迁移路径,探索草甘膦在岩溶地质中的吸附和去除能力,以更好地评估岩溶地质的碳汇稳定性和水生生态系统的安全性;(3)借助酶化学计量学等研究方法,探明草甘膦在土壤中的转化速率、通量以及与微生物代谢和营养需求、环境效应之间的耦合关系。     

荧光假单胞CLW17菌株对草甘膦的降解及其机制初探

探讨根际促生长细菌荧光假单胞菌(Pseudomonas fluorescens)CLW17对草甘膦的降解作用及其分子机制。利用平板培养法和钼锑抗比色法对CLW17菌株降解草甘膦能力进行研究。采用Plackett-Burman(PB)与Central Composite Design(CCD)试验联用确定CLW17降解草甘膦的最佳条件。对降解草甘膦关键基因thiO进行生物信息学分析,并构建敲除株(CLW17ΔthiO)及回补株(ΔthiO/thiO),研究其对草甘膦降解的影响。荧光假单胞菌CLW17菌株耐受草甘膦的最大浓度为0.7%。在降解草甘膦最优条件下,野生株CLW17、敲除株CLW17ΔthiO和回补株ΔthiO/thiO对草甘膦降解率分别为72.81%、32.78%和54.33%。生物信息学分析显示thiO基因编码366个氨基酸,拥有信号肽和跨膜结构域。CLW17具有较强的降解草甘膦能力,且thiO基因与降解草甘膦能力密切相关。研究结果可为草甘膦污染的治理及土壤修复提供良好的菌种资源。