Large-scale Analysis Of Differential Gene Expression In Coffee Genotypes Resistant And Susceptible To Leaf Miner-toward The Identification Of Candidate Genes For Marker Assisted-selection

Background: A successful development of herbivorous insects into plant tissues depends on coordination of metabolic processes. Plants have evolved complex mechanisms to recognize such attacks, and to trigger a defense response. To understand the transcriptional basis of this response, we compare gene expression profiles of two coffee genotypes, susceptible and resistant to leaf miner (Leucoptera coffella). A total of 22000 EST sequences from the Coffee Genome Database were selected for a microarray analysis. Fluorescence probes were synthesized using mRNA from the infested and non-infested coffee plants. Array hybridization, scanning and data normalization were performed using Nimble Scan® e ArrayStar® platforms. Genes with foldchange values +/-2 were considered differentially expressed. A validation of 18 differentially expressed genes was performed in infected plants using qRT-PCR approach.Results: The microarray analysis indicated that resistant plants differ in gene expression profile. We identified relevant transcriptional changes in defense strategies before insect attack. Expression changes (>2.00-fold) were found in resistant plants for 2137 genes (1266 up-regulated and 873 down-regulated). Up-regulated genes include those responsible for defense mechanisms, hypersensitive response and genes involved with cellular function and maintenance. Also, our analyses indicated that differential expression profiles between resistant and susceptible genotypes are observed in the absence of leaf-miner, indicating that defense is already build up in resistant plants, as a priming mechanism. Validation of selected genes pointed to four selected genes as suitable candidates for markers in assisted-selection of novel cultivars.Conclusions: Our results show evidences that coffee defense responses against leaf-miner attack are balanced with other cellular functions. 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Host colonization differences between citrus and coffee isolates of Xylella fastidiosa in reciprocal inoculation Diferenças em colonização do hospedeiro por isolados de Xylella fastidiosa de citros e cafeeiro em inoculações recíprocas

Citrus variegated chlorosis (CVC) and coffee stem atrophy (CSA) are important diseases in Brazil associated with closely-related strains of Xylella fastidiosa, but little is know about host aoverlappingnd importance of citrus and coffee as inoculum sources of these strains. In this study, reciprocal-inoculation experiments were performed to determine if CVC and CSA isolates are biologically similar within citrus and coffee plants. These two hosts were mechanically inoculated with a CVC and a CSA isolate of X. fastidiosa at four concentrations ranging between10³ and 10(9) colony forming units CFU mL-1. At two, four and eight months after inoculation, the infection efficiency and bacterial populations of the isolates in each host were determined by culturing. The CVC isolate infected both citrus and coffee plants, but developed lower populations in coffee. The CSA isolate did not colonize citrus. Inoculation of coffee plants with the CVC isolate resulted in low rates of infection and required an inoculum concentration ten-fold higher than that necessary to obtain a similar (25%) rate of infection in citrus. The relatively low infection rates and bacterial numbers of the CVC isolate in coffee plants compared with those observed in citrus suggest that coffee is not a suitable host to serve as a source of inoculum of the CVC strain for primary spread to citrus or within coffee plantations.<br>Clorose variegada dos citros (CVC) e atrofia dos ramos do cafeeiro (ARC) são doenças importantes no Brasil, associadas a estirpes de Xylella fastidiosa que são geneticamente próximas. Entretanto, pouco se sabe a respeito de plantas hospedeiras em comum e da importância de citros e cafeeiro como fontes de inóculo dessas estirpes. Neste estudo, realizaram-se experimentos de inoculação recíproca para determinar se isolados de X. fastidiosa de CVC e de ARC são biologicamente semelhantes em plantas de citros e café. Estes dois hospedeiros foram mecanicamente inoculados com um isolado de CVC e um isolado de ARC, em quatro concentrações que variaram de 10³ a 10(9) unidades formadoras de colônias UFC mL-1. Aos dois, quatro e oito meses após a inoculação, a eficiência de infecção e a população bacteriana dos isolados em cada hospedeiro foram determinadas por cultura. O isolado de CVC infectou tanto plantas de citros quanto de café, mas desenvolveu populações mais baixas em cafeeiro. O isolado de ARC não colonizou citros. A inoculação de plantas de café com o isolado de CVC resultou em baixas taxas de infecção e exigiu uma concentração de inóculo dez vezes mais alta que a necessária para obter uma taxa de infecção semelhante (25%) em citros. A reduzida taxa de infecção e a baixa população bacteriana do isolado de CVC em cafeeiro em relação a citros sugerem que cafeeiro não é um hospedeiro adequado para atuar como fonte de inóculo da estirpe de CVC para disseminação deste patógeno para pomares de laranja ou dentro de cafezais