Sugarcane yellow leaf disease in Brazil: Evidence of association with a luteovirus

Evidence of the viral etiology of sugarcane yellow leaf disease (SCYLD), occurring in southeast Brazil, was obtained by light and electron microscopy combined with serology. Light microscopy using epifluorescence illumination showed an abnormal yellow-green fluorescing material in the phloem of SCYLD-affected plants that was rarely observed in control plants. Immunolocalization in tissue-printed (or -blotted) nitrocellulose membranes, using barley yellow dwarf virus (BYDV) serotype PAV antiserum, showed a weak but clearly positive reaction in the phloem. Isometric viruslike particles of 24 to 26 nm in diameter were found by electron microscopy both in situ and in partially purified preparations. Examination of thin sections showed that phloem companion cells contained viruslike particles and presented cytological changes apparently related to the development of virus infection. Partially purified preparations produced UV absorption spectra typical of a nucleoprotein, with high absorbance at 260 nm, as expected for isometric virus particles. Virus particles were observed in extracts and partially purified preparations using immunosorbent electron microscopy with BYDV-PAV antiserum. Plate-trapped antigen enzyme-linked immunosorbent assays with the same antiserum indicate a weak serological relationship between BYDV-PAV and SCYLD-associated virus.811212

Identification of genes responsive to the application of ethanol on sugarcane leaves

The control of gene expression in precise time and space is a desirable attribute of chemically inducible systems. Ethanol is a chemical inducer with favourable features, such as being inexpensive and easy to apply. The aim of this study was to identify ethanol-responsive genes in sugarcane. The cDNA macroarray technique was adopted to identify transcript changes in sugarcane leaves (Saccharum spp. cv SP80-3280) exposed to ethanol. The expression profiles of sugarcane genes were analysed using nylon filters containing 3,575 cDNA clones from the leaf roll library of the SUCEST project. Seventy expressed sequence tags (ESTs) presented altered expression patterns, including ESTs corresponding to genes related to transcriptional and translational processes, abiotic stress and others. Several genes of unknown function were also identified. Among the 48 ESTs up-regulated by ethanol, an abiotic stress-responsive protein and an unknown function gene presented rapid induction by ethanol. The macroarray data of selected ethanol-responsive EST were confirmed by RNA-blot hybridisation. The expression profile of the 48 up-regulated genes was compared in two other cultivars: SP89-1115 and SP90-3414. Surprisingly, no gene showed a similar expression profile in the three cultivars. This result suggests that sugarcane plants have a high diversity in their responses to ethanol.26122119212

Survey in the sugarcane expressed sequence tag database (SUCEST) for simple sequence repeats

Sugarcane microsatellites or simple sequence repeats (SSR) were developed in an economical and practical way by mining EST databases. A survey in the SUCEST (sugarcane EST) database revealed a total of 2005 clusters out of 43 141 containing SSRs. Of these, 8.2% were dinucleotide, 30.5% were trinucleotide, and 61.3% were tetranucleotide repeats. Except for dinucleotides, the CG-rich motif types were the most common. Differences in abundance of trinucleotide motif types were observed between EST-SSRs and those isolated from sugarcane genomic libraries. Among the different cDNA libraries used for EST sequencing, SSRs were more frequent in the ones derived from leaf roll (LR). Twenty-three out of 30 tested SSRs produced scorable polymorphisms in 18 sugarcane commercial clones. These EST-SSRs showed a moderate level of polymorphism with some SSRs producing unique fingerprints. The number of alleles observed among the 18 clones evaluated varied from 2 to 15, with an average of 6.04 alleles/locus. The polymorphism information content (PIC) values ranged from 0.28 to 0.90 with a mean of 0.66. The EST-SSRs screened over both parents (SP 80-180; SP 80-4966) and 6 F-1 individuals produced 52 segregating markers that could potentially be used for sugarcane mapping. The EST-SSRs were found in clusters that had significant homology to proteins involved in important metabolic pathways such as sugar biosynthesis, proving that EST-SSRs are a valuable tool for the construction of a functional sugarcane map.47579580

Identification of new ABA- and MEJA-activated sugarcane bZIP genes by data mining in the SUCEST database

Sugarcane is generally propagated by cuttings of the stalk containing one or more lateral buds, which will develop into a new plant. The transition from the dormant into the active stage constitutes a complex phenomenon characterized by changes in accumulation of phytohormones and several other physiological aspects. Abscisic acid (ABA) and methyl-jasmonate (MeJA) are major signaling molecules, which influence plant development and stress responses. These plant regulators modulate gene expression with the participation of many transcriptional factors. Basic leucine zipper proteins (bZIPs) form a large family of transcriptional factors involved in a variety of plant physiological processes, such as development and responses to stress. Query sequences consisting of full-length protein sequence of each of the Arabidopsis bZIP families were utilized to screen the sugarcane EST database (SUCEST) and 86 sugarcane assembled sequences (SAS) coding for bZIPs were identified. cDNA arrays and RNA-gel blots were used to study the expression of these sugarcane bZIP genes during early plantlet development and in response to ABA and MeJA. Six bZIP genes were found to be differentially expressed during development. ABA and MeJA modulated the expression of eight sugarcane bZIP genes. Our findings provide novel insights into the expression of this large protein family of transcriptional factors in sugarcane.27233534

Functional integrated genetic linkage map based on EST-markers for a sugarcane (Saccharum spp.) commercial cross

The growing availability of ESTs provides a potentially valuable source of new DNA markers. The authors examined the SUCEST database and developed EST-derived markers. Thus to enhance the resolution of an existing linkage map and to identify putative functional polymorphic gene loci in a sugarcane commercial cross, 149 EST-SSRs and 10 EST-RFLPs were screened in the SP80-180 x SP80-4966 mapping population. With the markers already analyzed in the previous map, 2303 polymorphic markers were generated, of which 1669 (72.5%) were single-dose (SD) markers. Out of these 1669 SD markers, 664 (40%) were scattered onto 192 co-segregation groups (CGs) with a total estimated length of 6.261,1 cM. Using both genomic and EST-derived SSR and RFLP markers, 120 out of the 192 CGs were formed into fourteen putative homology groups (HGs). The EST-derived markers were subjected to BLASTX search in the SUCEST database, of which putative function was assigned to 113 EST-SSRs and six EST-RFLPs based on high nucleotide homology to previously studied genes. The integration of EST-derived markers improved the map, making it possible to consider additional fine mapping of the genome, and providing the means for developing 'perfect markers' associated with key QTL. To summarize, this paper deals with the construction of a genetic linkage map of sugarcane that is populated by functionally associated markers.20318920

Development of an integrated genetic map of a sugarcane (Saccharum spp.) commercial cross, based on a maximum-likelihood approach for estimation of linkage and linkage phases

Sugarcane (Saccharum spp.) is a clonally propagated outcrossing polyploid crop of great importance in tropical agriculture. Up to now, all sugarcane genetic maps had been developed using either full-sib progenies derived from interspecific crosses or from selfing, both approaches not directly adopted in conventional breeding. We have developed a single integrated genetic map using a population derived from a cross between two pre-commercial cultivars ('SP80-180' x 'SP80-4966') using a novel approach based on the simultaneous maximum-likelihood estimation of linkage and linkage phases method specially designed for outcrossing species. From a total of 1,118 single-dose markers (RFLP, SSR and AFLP) identified, 39% derived from a testcross configuration between the parents segregating in a 1:1 fashion, while 61% segregated 3:1, representing heterozygous markers in both parents with the same genotypes. The markers segregating 3:1 were used to establish linkage between the testcross markers. The final map comprised of 357 linked markers, including 57 RFLPs, 64 SSRs and 236 AFLPs that were assigned to 131 co-segregation groups, considering a LOD score of 5, and a recombination fraction of 37.5 cM with map distances estimated by Kosambi function. The co-segregation groups represented a total map length of 2,602.4 cM, with a marker density of 7.3 cM. When the same data were analyzed using JoinMap software, only 217 linked markers were assigned to 98 co-segregation groups, spanning 1,340 cM, with a marker density of 6.2 cM. The maximum-likelihood approach reduced the number of unlinked markers to 761 (68.0%), compared to 901 (80.5%) using JoinMap. All the co-segregation groups obtained using JoinMap were present in the map constructed based on the maximum-likelihood method. Differences on the marker order within the co-segregation groups were observed between the two maps. Based on RFLP and SSR markers, 42 of the 131 co-segregation groups were assembled into 12 putative homology groups. Overall, the simultaneous maximum-likelihood estimation of linkage and linkage phases was more efficient than the method used by JoinMap to generate an integrated genetic map of sugarcane.112229831

Transcriptionally active transposable elements in recent hybrid sugarcane

Transposable elements (TEs) are considered to be important components of the maintenance and diversification of genomes. The recent increase in genome sequence data has created an opportunity to evaluate the impact of these active mobile elements on the evolution of plant genomes. Analysis of the sugarcane transcriptome identified 267 clones with significant similarity to previously described plant TEs. After full cDNA sequencing, 68 sugarcane TE clones were assigned to 11 families according to their best sequence alignment against a fully characterized element. Expression was further investigated through a combined study utilizing electronic Northerns, macroarray, transient and stable sugarcane transformation. Newly synthesized cDNA probes from flower, leaf roll, apical meristem and callus tissues confirm previous results. Callus was identified as the tissue with the highest number of TEs being expressed, revealing that tissue culture drastically induced the expression of different elements. No tissue-specific family was identified. Different representatives within a TE family displayed differential expression patterns, showing that each family presented expression in almost every tissue. Transformation experiments demonstrated that most Hopscotch clone-derived U3 regions are, indeed, active promoters, although under a strong transcriptional regulation. This is a large-scale study about the expression pattern of TEs and indicates that mobile genetic elements are transcriptionally active in the highly polyploid and complex sugarcane genome.44570771