Expressão diferencial dos microRNAs miR319 e miR397 em cana-de-açúcar infectada por Xanthomonas albilineans
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2017-05-03
Autores
Santos, Thiago Mateus Rosa dos [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A cana-de-açúcar é acometida por uma doença conhecida por “escaldadura das folhas” causada pela bactéria colonizadora do xilema Xanthomonas albilineans, considerada uma das principais doenças que atingem a cultura da canade-açúcar. A sintomatologia na fase crônica se caracteriza, principalmente, pelo aparecimento de uma faixa branca ao lado da nervura central da folha, a qual evolui para clorose total causando a morte da planta. Uma vez que o patógeno pode ser transmitido de várias maneiras, o seu controle demanda altos custos. Desta maneira, o desenvolvimento de cultivares tolerantes é uma boa opção para o controle efetivo da doença. A tolerância e sensibilidade das plantas aos fatores bióticos está relacionada com a expressão de genes, e dentre estes, os miRNAs (incluindo o miR397 e o miR319) têm sido relatados como importantes reguladores em vários mecanismos de resposta das plantas. O objetivo deste trabalho foi analisar a expressão de dois miRNAs (miR319 e miR397) em duas cultivares de cana-de-açúcar (RB86-7515 - tolerante e SP78-4467 - suscetível), infectadas por uma linhagem de X. albilineans (IACXa11), considerada a mais virulenta do Brasil. Para isto, as plantas foram cultivadas em vasos, inoculadas com X. albilineans e mantidas em casa de vegetação. Amostras de folhas e colmos foram coletadas em cinco períodos (24, 72, 144, 360 e 720 h) e a expressão dos miRNAs foi analisada pela técnica de Stem-loop RT-qPCR. Os miR397 e miR319 apresentaram-se diferencialmente expressos nas cultivares e entre os tecidos. Na cultivar suscetível (SP78-4467), durante os primeiros períodos de infecção (24, 72 e 144 h), houve uma resposta tardia de defesa quando comparada com a cultivar tolerante (RB86-7515). O miR319 apresentou o mesmo perfil de expressão em folhas e colmos da cultivar RB86-7515 (tolerante), sugerindo que o reconhecimento do patógeno e a ativação dos mecanismos de defesa são modulados em ambos os tecidos. De maneira geral, as análises dos miRNAs demonstraram que a expressão do miR397 é menor quando comparada com o miR319. O mesmo padrão foi observado para os seus respectivos genes alvo. O miR397 regula a enzima lacase, importante na biossíntese de lignina. A repressão deste miRNA aumentaria a lignificação, sugerindo um mecanismo estrutural de resposta. O miR319 regula os fatores de transcrição (FTs) MYB e TCP, os quais são responsáveis pela sinalização de ácido abscísico (ABA) e ácido jasmônico (JA). A repressão destes hormônios vegetais desencadeia a sinalização por ácido salicílico (SA), o qual é responsável pela defesa contra patógenos hemibiotróficos, tal como X. albilineans.
Sugarcane is affected by a disease known as "leaf scald" caused by the bacterium Xanthomonas albilineans, which colonizes the xylem. This disease is one of the most important for sugarcane culture. The chronic phase is mainly characterized by the white band emergence along the central leaf vein, which causes total chlorosis of the leaf and plant death. Since the pathogen can be transmitted in many ways, his control demands high costs, and the development of tolerant cultivars is a good option for disease control. The plant tolerance and sensitivity to biotic factors is related to gene expression, and among these, the miRNAs (including miR397 and miR319), have been reported as important regulators in various plant response mechanisms. The aim of this work was to analyze the expression of two miRNAs (miR319 and miR397) in two sugarcane cultivars (RB86-7515 – tolerant, and SP78- 4467 - susceptible), infected by a strain of X. albilineans (IACXa11), the most virulent in Brazil. The plants were grown in vases, inoculated with X. albilineans and kept in a greenhouse. Samples of leaves and stems were collected in five periods (24, 72, 144, 360, and 720 h), and the miRNA expression was analyzed by Stem-loop RT-qPCR. The miR397 and miR319 expression were different between cultivars and tissues. In the susceptible cultivar (SP78-4467), during the first infection periods (24, 72 and 144 h), there was a late defense response when compared to the tolerant cultivar (RB86-7515). The miR319 presented the same expression profile in leaves and stems of the cultivar RB86-7515 (tolerant), suggesting that the pathogen recognition and defense mechanisms activation were modulated in both tissues. In general, miRNAs analyzes demonstrated that miR397 expression is lower when compared to miR319. The same pattern was observed for their respective target genes. The miR397 is a laccase regulator, important in lignin biosynthesis. Repression of this miRNA would increase lignification, suggesting a structural mechanism of response. The miR319 regulates the transcription factors (TFs) MYB and TCP, which are responsible for the abscisic acid (ABA) and jasmonic acid (JA) signaling. The repression of these plant hormones triggers salicylic acid (SA) signaling pathway, which is responsible for the defense against hemibiotrophic pathogens, such as X. albilineans.
Sugarcane is affected by a disease known as "leaf scald" caused by the bacterium Xanthomonas albilineans, which colonizes the xylem. This disease is one of the most important for sugarcane culture. The chronic phase is mainly characterized by the white band emergence along the central leaf vein, which causes total chlorosis of the leaf and plant death. Since the pathogen can be transmitted in many ways, his control demands high costs, and the development of tolerant cultivars is a good option for disease control. The plant tolerance and sensitivity to biotic factors is related to gene expression, and among these, the miRNAs (including miR397 and miR319), have been reported as important regulators in various plant response mechanisms. The aim of this work was to analyze the expression of two miRNAs (miR319 and miR397) in two sugarcane cultivars (RB86-7515 – tolerant, and SP78- 4467 - susceptible), infected by a strain of X. albilineans (IACXa11), the most virulent in Brazil. The plants were grown in vases, inoculated with X. albilineans and kept in a greenhouse. Samples of leaves and stems were collected in five periods (24, 72, 144, 360, and 720 h), and the miRNA expression was analyzed by Stem-loop RT-qPCR. The miR397 and miR319 expression were different between cultivars and tissues. In the susceptible cultivar (SP78-4467), during the first infection periods (24, 72 and 144 h), there was a late defense response when compared to the tolerant cultivar (RB86-7515). The miR319 presented the same expression profile in leaves and stems of the cultivar RB86-7515 (tolerant), suggesting that the pathogen recognition and defense mechanisms activation were modulated in both tissues. In general, miRNAs analyzes demonstrated that miR397 expression is lower when compared to miR319. The same pattern was observed for their respective target genes. The miR397 is a laccase regulator, important in lignin biosynthesis. Repression of this miRNA would increase lignification, suggesting a structural mechanism of response. The miR319 regulates the transcription factors (TFs) MYB and TCP, which are responsible for the abscisic acid (ABA) and jasmonic acid (JA) signaling. The repression of these plant hormones triggers salicylic acid (SA) signaling pathway, which is responsible for the defense against hemibiotrophic pathogens, such as X. albilineans.
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Palavras-chave
Estresse biótico, miRNA, miR319, miR397, Saccharum officinarum, Xanthomonas albilineans, Biotic stress