Does the Glyphosate Treatment Interfere Negatively on RNA Integrity in Glyphosate-Resistant and -Sensitive Conyza bonariensis?

Abstract

The hairy fleabane (Conyza bonariensis (L.) Cronq.) is among the most problematic glyphosate-resistant weeds to manage around the world. In weed science, molecular approaches such as RNA sequencing (RNA-Seq) and quantitative reverse-transcription polymerase chain reaction (RT-qPCR) have been employed to study molecular responses to glyphosate treatment in Conyza species. Glyphosate treatment leads to reactive oxygen species (ROS) production in plants which could damage the RNA. Degraded RNA is an issue and can compromise further molecular analysis. Thus, the objective of this study was to evaluate whether glyphosate treatment interferes negatively on RNA integrity of glyphosate-resistant andsensitive hairy fleabane biotypes. Two experiments were performed using glyphosate doses from 0 to 11,840 g a.e. ha -1 and evaluated in a time-course until 288 hours after treatment. The total of 86 RNA samples were evaluated. The RNA integrity was evaluated in a Bioanalyzer 2100 equipment according to RNA integrity number (RIN) scores and electrophoresis gel. The RIN scores ranged from 5.1 to 9.0. Glyphosate doses do not reduce the RIN scores in both glyphosate-resistant and -sensitive biotypes of hairy fleabane. Visual and automatic analysis of electrophoresis gel show suitable results for all RNA samples, with well-defined bands at 28S and 18S positions and no degradation. The results of the analysis indicate that glyphosate treatment does not affect the RNA integrity of glyphosate-resistant and -sensitive biotypes of hairy fleabane until 288 and 192 hours after glyphosate treatment, respectively. The RNA integrity analysis provides useful results to evaluate the RNA condition for further analysis. However, the costs were around US$ 14.25 per sample, considering only reagents. These results are useful for planning future timecourse experiments in Conyza spp. after glyphosate treatment.Keywords: hairy fleabane, molecular analysis, RNA integrity number (RIN). (Conyza bonariensis (L.) Cronq.) está entre as plantas daninhas resistentes ao glifosato mais difíceis de serem manejadas em todo o mundo. Na ciência das plantas daninhas, abordagens moleculares, como o sequenciamento de RNA (RNA-Seq) e a reação da transcriptase reversa da polimerase em tempo real (RT-qPCR), têm sido empregadas para estudar as respostas moleculares ao tratamento com glifosato em espécies de Conyza. No entanto, o tratamento com glifosato leva à produção de espécies reativas de oxigênio (ROS) em plantas, que podem danificar o RNA. A degradação do RNA é um problema e pode comprometer futuras análises moleculares. Assim, o objetivo deste estudo foi avaliar se o tratamento com glifosato interfere negativamente na integridade do RNA de biótipos de buva resistente e sensível ao glifosato. Dois experimentos foram realizados I D I D I D Planta Daninha 2019; v37:e019217909 PIASECKI, C. et al. Does the glyphosate treatment interfere negatively on RNA integrity in glyphosate-resistant and -sensitive ... 2 utilizando doses de glifosato de 0 a 11.840 g e.a. ha -1 e av...