RESUMO
Several molecular tools have been used to clarify the basis of plant-bacteria interaction; however, the mechanism behind the association is still unclear. In this study, we used a proteomic approach to investigate the root proteome of Zea mays (cv. DKB240) inoculated with Herbaspirillum seropedicae strain SmR1 grown in vitro and harvested 7 days after inoculation. Eighteen differentially accumulated proteins were observed in root samples, ten of which were identified by MALDI-TOF mass spectrometry peptide mass fingerprint. Among the identified proteins, we observed three proteins present exclusively in inoculated root samples and six upregulated proteins and one downregulated protein relative to control. Differentially expressed maize proteins were identified as hypothetical protein ZEAMMB73_483204, hypothetical protein ZEAMMB73_269466, and tubulin beta-7 chain. The following were identified as H. seropedicae proteins: peroxiredoxin protein, EF-Tu elongation factor protein, cation transport ATPase, NADPH:quinone oxidoreductase, dinitrogenase reductase, and type III secretion ATP synthase. Our results presented the first evidence of type III secretion ATP synthase expression during H. seropedicae-maize root interaction.
Assuntos
Proteínas de Bactérias/genética , Herbaspirillum/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/microbiologia , Proteômica , Zea mays/genética , Zea mays/microbiologia , Endófitos/metabolismo , Endófitos/fisiologia , Expressão Gênica , Herbaspirillum/metabolismoRESUMO
The present study aimed to identify salinity-tolerant genes in three cultivars (BRS-7 Taim, BRS Querência and BRS Atalanta) of Oryza sativa L. ssp. indica S. Kato and in three cultivars (BRS Bojurú, IAS 12-9 Formosa and Goyakuman) of Oryza sativa L. ssp. japonica S. Kato. Ten days after emergence seedlings were transferred to a greenhouse and placed in a 15L vessel with half strength Hoagland nutrient solution, which was changed every four days, under controlled temperature and humidity. Plants were harvested 56 days after transfer. DNA extraction was carried out by CTAB method and salinity-tolerant genes SOS and CK1 were identified by in silico research. Amplification of gene sequence was performed with in silico primers. Bands were detected by agar gel electrophoresis and visualized under ultraviolet light after staining with ethidium bromide. Gene SOS1 fragments were present in all cultivars, except in BRS Atalanta, whereas CK1 gene was present in all evaluated cultivars. Results show that salinity-tolerant genes under analysis were identified in the two sub-species.
O estudo teve como objetivo identificar genes envolvidos na tolerância à salinidade em três cultivares (BRS-7 Taim, BRS Querência e BRS Atalanta) de Oryza sativa L. ssp. indica S. Kato e em três cultivares (BRS Bojurú, IAS 12-9 Formosa e Goyakuman) de Oryza sativa L. ssp. japonica S. Kato. As plântulas foram transferidas para casa de vegetação, sob temperatura e umidade relativa controladas, dez dias após a emergência e colocadas em bacia com capacidade para 15 litros, contendo solução nutritiva de Hoagland, meia força, a qual foi mudada em intervalos de quatro dias. A coleta foi aos 56 dias após a transferência. A extração de DNA foi realizada pelo método CTAB. Os genes SOS e CK1, envolvidos na tolerância à salinidade, foram identificados por meio de pesquisa in sílico . A amplificação das sequências do gene foi realizada utilizando-se primers também desenhados in sílico. As bandas foram detectadas por eletroforese em gel de agarose e visualizadas em luz ultravioleta após coloração com brometo de etídio. Fragmentos do gene SOS1 foram encontrados em todas as cultivares, exceto para BRS Atalanta. O gene CK1 esteve presente em todas as cultivares avaliadas. Os resultados mostram que os genes estudados estão presentes em ambas as subespécies.