Search

Sugarcane orange rust, caused by the fungus Puccinia kuehnii, is a serious disease of sugarcane. The most effective strategy to combat the disease is to develop resistant cultivars. Phenotypic screening for resistance is laborious, and breeders would benefit from molecular markers linked to the trait. The objective of this research was to identify, via association mapping, markers linked to resistance to orange rust. From the germplasm collection at Canal Point, Florida, 724 genotypes were screened for orange rust resistance via artificial inoculation of field-grown plants. A total of 38 susceptible and 37 resistant genotypes that were at or near the extremes of the population distribution were chosen for genotyping. Genotyping was accomplished via capture sequencing using 32,555 sugarcane probes anchored to the sorghum genome. Several methods were used to account for population structure and/or kinship. The most appropriate methods (determined by the genomic inflation factor) were mixed models and EIGENSTRAT. A total of 38 unique SNPs identified by these methods were then validated on 560 members of the population, using high resolution melting of PCR amplicons. Ten of these markers were statistically significant for the quantitative measure of rust severity, of which nine were also significant for the qualitative measure of ‘status’ (resistant/susceptible). The maximum amount of variation explained by any single marker was approximately 11%. This research has provided markers which can be deployed to help select superior parents for orange rust resistance, and also provided valuable insight for future research towards genetic control of orange rust in sugarcane.

In September 2007 at Masagua, Escuintla Department, Guatemala, uredial lesions that appeared different from those of brown rust were observed on a sugarcane (a complex hybrid of Saccharum L. species) cultivar (CP 72-2086) considered resistant to brown rust caused by Puccinia melanocephala Syd. & P. Syd. Samples were sent to the USDA-ARS Systematic Mycology and Microbiology Laboratory in Beltsville, MD for identification. Observed morphological features were consistent with P. kuehnii E.J. Butler and appeared similar to orange rust samples obtained from Florida in July (2). Uredinial lesions were hypophyllous, orange, and variable in size measuring 650 to 850 × 26 to 32 μm. Urediniospores were mostly obovoid to pyriform or broadly ellipsoidal, variable in size, 32 to 45 × 25 to 30 μm, and moderately echinulate with spines evenly distributed, 3 to 5 μm apart. Urediniospore walls were orange-to-light cinnamon brown, 1 to 2.5 μm thick with a pronounced apical wall and four to five equatorial pores. Telia and teliospores were not observed. The nuclear large subunit rDNA region of the rust infecting cv. CP 72-2086 (BPI 898289, GenBank Accession No. EU344904) and the ITS1, 5.8S, and ITS2 rDNA regions (GenBank Accession No. EU543434) were sequenced (1,3). DNA sequences matched sequences of P. kuehnii in GenBank and were distinct from known sequences of P. melanocephala available in GenBank (3). Thirteen cultivars were rated as to their relative resistance using severity of orange rust symptoms; CG 96–59, CG 96–135, CP 72–1312, CP 73–1547, and CP 88–1165 were resistant; CG 96–40, CG 98–121, CP 72–2086, CP 88–1508, and CP 89–2143 were intermediate; and CG 96–52, CG 98–0115, and SP 79–2233 were susceptible. Orange rust was previously reported in Florida (2), but to our knowledge, this is the second report of its occurrence in the Western Hemisphere. References: (1) M. C. Aime. Mycoscience 47:112, 2006. (2) J. C. Comstock et al. Plant Dis. 92:175, 2008. (3) E. V. Virtudazo et al. Mycoscience 42:447, 2001.

Symptoms and signs of orange rust on sugarcane (a complex hybrid of Saccharum L. species) were observed from July 2007 on cv. SP 71-5574 in Costa Rica at the Coopeagri Sugar Mill located in Pérez Zeledón, San José and on multiple cultivars (CP 72-2086, Pindar, Q 132, Q 138, SP 71-5574, and SP 79-2233) at the Providencia Sugar Mill near Muelle, San Carlos and Cutris Sugar Mill near Los Chiles during August 2007. The same symptoms and signs were observed on cv. CP 72-2086 during September 2007 in Nicaragua at Ingenio San Antonio, located near Chinandega, and Ingenio Monte Rosa near El Viejo, Nicaragua. Disease symptoms and uredinia appeared different from brown rust caused by Puccinia melanocephala, and brown rust usually does not occur on these cultivars. Uredinia and urediniospores were similar to those described for orange rust (1,2). Cvs. SP 71-5574 and SP 79-2233 are susceptible and cv. CP 72-2086 is moderately susceptible to orange rust in Costa Rica and cvs. ISACP 00-1075, ISA 00-1000, and CP 72-2086 are moderately susceptible in Nicaragua. Samples from both locations (Costa Rica BPI No. 878816 and Nicaragua BPI No. 878817) examined at the USDA-ARS Mycology and Microbiology Laboratory in Beltsville, MD showed morphological characteristics consistent with those of P. kuehnii. Analysis of ITS1, 5.8S, and ITS2 rDNA sequences of the rust infecting cv. CP 72-2086 (GenBank Accession No. FJ532477) from Costa Rica and cv. ISA 00-1000 from Nicaragua (GenBank Accession No. FJ532476) confirmed the identity as P. kuehnii, the causal agent of sugarcane orange rust. Beside the cultivars already mentioned, orange rust also was confirmed on cvs. RB 73-9735 and CPCL 02-2130 in Costa Rica. To our knowledge, this is the first report of orange rust of sugarcane in Costa Rica and Nicaragua and the third confirmation of the disease in the Western Hemisphere and Caribbean Basin. References: (1) J. C. Comstock et al. Plant Dis. 92:175, 2008. (2) W. Ovalle et al. Plant Dis. 92:973, 2008.

Sood, S. G., Comstock, J. C., and Glynn, N. C. 2009. Leaf whorl inoculation method for screening sugarcane rust resistance. Plant Dis. 93:1335-1340. Brown rust, caused by Puccinia melanocephala, and orange rust, caused by P. kuehnii, are agronomically important diseases of sugarcane in Florida. Cultivar resistance is the best means of controlling these diseases. Natural infection has been the primary means of assessing resistance in sugarcane cultivars against rusts; unfortunately, natural infection is not always efficient in identifying resistant cultivars due to variable environmental conditions. Therefore, a more reliable screening method is needed to effectively select resistant genotypes. An inoculation technique was evaluated for identification of brown and orange rust resistance in sugarcane cultivars. Inoculations were performed in the field by placing a 0.5-ml urediniospore suspension in the leaf whorl of three individual sugarcane stalks per plant using a pipette. Symptoms developed on leaves of all the susceptible cultivars after 4 weeks, and appeared as a band of pustules. Plants were rated for their reaction to rust 4 weeks after inoculation. The optimum concentrations of inoculum for expression of brown and orange rust symptoms were determined. The most severe brown rust and orange rust symptoms were observed using inoculum containing 10 5 and 104 urediniospores/ml, respectively. Clones in several stages of the Canal Point breeding program were screened for their rust reaction by leaf whorl inoculation. The technique enabled rapid screening of a large number of cultivars in field plantings using a small amount of inoculum and limited man hours.

Symptoms of sugarcane orange rust were observed on July 17, 2008 on sugarcane cvs. Mex 57-1285, Mex 61-230, and Co 301 (a clone received in Mexico in 1953) at the Centro de Investigación y Desarrollo de la Caña de Azúcar en Tuxtla Chico, Chiapas, Mexico. In El Salvador, from August 2008 through January 2009, rust symptoms were observed on cv. CP 72-2086 (previously resistant to brown rust caused by Puccinia melanocephala Syd. & P. Syd.) in 117 dispersed sugarcane-production fields in various localities of El Salvador. Likewise, rust symptoms were first observed on sugarcane cv. SP 74-8355 (more than 25% severity and considered resistant to brown rust) at Natá, Coclé Province in Panama from January to February 2008. Dried herbarium leaf samples of sugarcane rust-infected leaves collected in El Salvador and Mexico were sent to the ARS, USDA Systematic Mycology and Microbiology Laboratory in Beltsville MD for identification. Panamanian samples were collected similarly and analyzed at the CALESA Biotechnology Laboratory. Morphological features of uredinial lesions and urediniospores were distinct from those of P. melanocephala and consistent with P. kuehnii E. J. Butler observed previously on specimens from Florida, Guatemala, Costa Rica, and Nicaragua (1–3). Analysis of the ITS1, 5.8S, and ITS2 and 28S large subunit rDNA sequences of the rust on infected cvs. Mex 57-1285, Mex 61-230, and Co 301 (BPI 878930, 879139, and 879140; GenBank Accession Nos. GO283006, GO283004, and GO283005, respectively) from Mexico and cv. CP 72-2086 from three locations in El Salvador (BPI 879135, 879136, and 879137; GenBank Accession Nos. GO283009, GO283007, and GO283008, respectively) all confirmed the identification of P. kuehnii. Similar analysis of the ITS1, 5.8S, and ITS2 rDNA sequence for the rust infecting cv. SP 74-8355 (GenBank Accession No. GO281584) confirmed the identification of P. kuehnii in Panama. To our knowledge, this is the first report of P. kuehnii causing orange rust disease of sugarcane in El Salvador, Mexico, and Panama. These findings also confirm the wider distribution of orange rust in the Western Hemisphere. References: (1) E. Chavarria et al. Plant Dis. 93:425, 2009. (2) J. C. Comstock et al. Plant Dis. 92:175, 2008. (3) W. Ovalle et al. Plant Dis. 92:973, 2008.

Orange rust of sugarcane (Saccharum spp. hybrids), caused by Puccinia kuehnii, is a relatively new disease in the Western Hemisphere that substantially reduces yields in susceptible sugarcane genotypes. The objective of this study was to determine the physiological mechanisms of orange rust–induced reductions in sugarcane growth and yield by quantifying effects of the disease on leaf SPAD index (an indication of leaf chlorophyll content), net photosynthetic rate, dark respiration, maximum quantum yield of CO2 assimilation, carbon fixation efficiency, and the relationships between these leaf photosynthetic components and rust disease ratings. Plants growing in pots were inoculated with the orange rust pathogen using a leaf whorl inoculation method. A disease rating was assigned using a scale from 0 to 4 with intervals of 0.5. At disease ratings ≥2, the rust-infected leaf portion of inoculated plants showed significant reductions in SPAD index, maximum quantum yield, carbon fixation efficiency, stomatal conductance, leaf transpiration rate, and net photosynthetic rate; but the rusted portion of the infected leaves had increased intercellular CO2 concentration and leaf dark respiration rate. Although leaf SPAD index, photosynthetic rate, stomatal conductance, and transpiration rate at the rust-infected portion decreased linearly with increased rust rating, the effect of orange rust on photosynthetic rate was much greater than that on stomatal conductance and transpiration. Unlike earlier reports on other crops, reduction in leaf photosynthesis by orange rust under low light was greater than that under high light conditions. These results help improve the understanding of orange rust etiology and physiological bases of sugarcane yield loss caused by orange rust.

Background Neonatal Intensive Care (NICU) patients have individual nutritional requirements often requiring Patient Specific Parenteral Nutrition (PSPN). From October 2015, the national PSPN compounding service availability changed from 7 days per week service to 5 days per week (i.e. no weekend and limited bank holiday ordering available). The aim of this study was to examine the introduction of a 5 day only PSPN supply on neonatal patient parenteral nutrition availability in a tertiary NICU. Methods We performed a prospective assessment of the provision of a 5 day rather than 7 day ordering of PSPN over a one month period (June 2017). Results Fifteen neonatal patients received a cumulative 89 days of PN during June 2017. 10 (66%) patients received PSPN during this time period. There was same day availability of PSPN in 62 of 89 days of PN (69%). Conclusion Thorough education and training will help prescribers to make appropriate PSPN ordering decisions. Improvements to available stock bags may reduce the amount of PSPN that is required but a 7 day PSPN ordering service would improve efficient provision of clinically indicated PSPN to premature infants in NICU in Ireland.

Among the species related to sugarcane, Saccharum spontaneum (L.) is a wild species with the greatest potential as a source of genetic variation to cope with biomass production in harsh environments. Due to its high yield, early vigor, ratooning ability, low input requirements and tolerance to various biotic and abiotic stresses, sugarcane breeders have shown interest in its contribution, as a donor of genes, to the development of high biomass energy canes. The conservation, evaluation and utilization of the genetic variability of S. spontaneum available in germplasm collections are critical for breeding, but, given the aggressive rhizomatous growth habit and the ability to propagate via seed dispersal, S. spontaneum is classified as a noxious weed in several nations, including the U.S.A. As a result, field trials are restrictive and few phenotypic analyses have been carried out on these collections. In the present study, a subset of 130 S. spontaneum accessions obtained from the World Collection of Sugarcane and Related Grasses in Miami, FL has been characterized phenotypically - with either morphological and agronomic traits (including composition analysis) or reaction to abiotic stress and genotypically - molecular markers (Simple Sequence Repeats - SSR). Using these data, a core collection has been established, genotypes with positive agronomic traits have been identified and are being used as parents for hybridization crosses, aimed at genetic improvement of sugarcane and energycane.

Sugarcane (Saccharum spp.) is an important economic crop for producing edible sugar and bioethanol. Brown rust has long been a major disease impacting sugarcane production worldwide. Resistance resource and markers linked to resistance are valuable tools for disease resistance improvement. An F1 segregating population derived from a cross between two hybrid sugarcane clones, brown rust-susceptible CP95-1039 and brown rust-resistant CP88-1762, were genotyped using genotyping by sequencing approach and also phenotyped in a replicated field trial. Single nucleotide polymorphism (SNP) and presence/absence markers were called with seven different pipelines to maximize reliable marker identification. High-density maps were constructed for both parental clones with a total map length of 4224.4 cM, and a marker density of one marker per 1.7 cM for CP95-1039, and a total map length of 4373.2 cM, and one marker per 2.0 cM for CP88-1762. Among the seven SNP callers, Tassel and Genome Analysis ToolKit performed better than other callers in single-dose SNP detection and contribution to genetic maps. Two major quantitative trait loci (QTL) controlling brown rust resistance were identified, which can explain 21 and 30% of the phenotypic variation, respectively. The genetic maps generated here will improve our understanding of sugarcane’s complex genome structure and discovery of underlying sequence variations controlling agronomic traits. The putative QTL controlling brown rust resistance can effectively be utilized in sugarcane breeding programs to expedite the selection process of brown rust resistance after validation.

The Saccharum L. genus includes important crops that are utilized for sugar and fuel production. The World Collection of Sugarcane and Related Grasses (World Collection) in Miami, FL contains diverse and potentially useful germplasm for this and related genera; however, this collection has been underutilized because little is known about the traits of its accessions. Our objectives were to phenotypically characterize the World Collection and select a representative core collection that could then be studied intensively. In total, eight morphological traits of the World Collection were evaluated three times in 1 year. A core of 300 accessions that included each species in the World Collection was selected by using the Maximization Strategy in MStrat software. The core had a higher diversity rating than random selections of 300 accessions. The Shannon–Weaver Diversity Index scores of the core and whole collection were similar indicating that the majority of the diversity was captured by the core collection. The ranges and medians between the core and World Collection were similar; only two of the trait medians were not significant at P = 0.05 using the non-parametric Wilcoxon method and the coincidence rate (CR % = 96.2) was high (>80) indicating that extreme values were retained. Thus, the phenotypic diversity of these traits in the World Collection was well represented by the core collection. Agronomic studies on the core should be useful for characterizing the World Collection and genes for useful traits should be available in the core collection.

‘CPCL 05-1102’ (Reg. No. CV-157, PI 667556) sugarcane (a complex hybrid of spp.) is the product of research initiated by the United States Sugar Corporation (USSC) and completed cooperatively by the USDA-ARS, the University of Florida, and the Florida Sugar Cane League, Inc. CPCL 05-1102 was released to growers in Florida on 16 Oct. 2012. CPCL 05-1102 was selected from a cross between the USSC proprietary cultivars CL 89-5189 (female) and CL 88-4730 (male) made at Clewiston, FL on 18 Dec. 2000. CL 89-5189 was adapted to muck soils, where it was used only briefly in commercial plantings of USSC due to yield losses to ratoon stunt (caused by subsp.). CL 88-4730 is widely used on commercial plantings of USSC, particularly on sand soils. CPCL 05-1102 has acceptable resistance to leaf scald (caused by Ashby, Dowson), strain E (mosaic), smut (caused by Sydow & P. Sydow), orange rust (caused by E.J. Butler), and brown rust (caused by H. & P. Sydow). 1, a major gene for resistance to brown rust, was not detected in the DNA of CPCL 05-1102. CPCL 05-1102 has high cane and sucrose yields and excellent freeze tolerance and is expected to be used on muck soils in Florida.

Ambient seismic noise is characterized by randomness incurred by the random position and strength of the noise sources as well as the heterogeneous properties of the medium through which it propagates. Here we use ambient noise data recorded prior to the 1996 Gjálp eruption in Iceland in order to show that a reduction of noise randomness can be a clear short-term precursor to volcanic activity. The eruption was preceded on 29 September 1996 by a Mw ~5.6 earthquake that occurred in the caldera rim of the Bárdarbunga volcano. A significant reduction of randomness started occurring 8 days before the earthquake and 10 days before the onset of the eruption. This reduction was observed even at stations more than 100 km away from the eruption site. Randomness increased to its previous levels 160 minutes after the Bárdarbunga earthquake, during which time aftershocks migrated from the Bárdarbunga caldera to a site near the Gjálp eruption fissure. We attribute this precursory reduction of randomness to the lack of higher frequencies (>1 Hz) in the noise wavefield caused by high absorption losses as hot magma ascended in the upper crust.

Leaf refl ectance has been used to estimate crop leaf chemical and physiological characteristics. Sugarcane (Saccharum spp.) leaf N, C, and chlorophyll levels are important traits for high yields and perhaps useful for genotype evaluation. The objectives of this study were to identify sugarcane genotypic variation in leaf hyperspectral refl ectance, leaf chemical (chlorophyll, N, and C), and yield (juice sucrose content, commercial recoverable sucrose [CRS], cane yield in tonnes of cane per hectare [TCH], and sucrose yield in tonnes of sucrose per hectare [TSH]) traits and to determine relationships between leaf refl ectance and these chemical and yield traits. In Stage II of the Canal Point, FL, sugarcane cultivar development program, we measured spectral refl ectance and chemical traits on three leaves of the top visible dewlap from each of 87 genotypes in December 2008 and 208 and 124 genotypes in May, July, September, and October 2009 and 2010, respectively. Yield traits were determined on mature plants. Genotypic variation of leaf refl ectance mainly occurred in 540 to 1200 nm. Leaf relative chlorophyll, N, C, and C to N ratio varied among genotypes and during the growing season. Highly signifi cant calibrations were developed for leaf chemical traits using leaf refl ectance (P < 0.0001). Correlations of juice sucrose, CRS, TCH, and TSH with leaf refl ectance were poor. Measurement of leaf refl ectance is a promising tool for estimating leaf chemical traits but not for predicting yield traits across a large number of diverse genotypes in early selection stages of a sugarcane breeding program.

A better understanding of sugarcane (Saccharum spp.) genetic variability in agronomic performance will help optimize breeding and selection strategies. Vigor ratings and Brix data were collected from the 2009 and 2010 clones in the first clonal selection stage (stage I) of the Canal Point (CP) sugarcane cultivar development program. Stage I individual selection was based on disease resistance and on the product of vigor and Brix. Vigor ratings (from 1 to 9) from all clones and Brix of any clones with a vigor rating ≥6 were collected in the stage I fields and analyzed for relationships between vigor and Brix, for selection rate in each family (i.e., cross), and for their coefficients of variation (CV) within and among families. There was no correlation between vigor and Brix, suggesting that it would be feasible in stage I to select sugarcane clones with both high vigor and high Brix. Variability was high (CV = 59%) for both the number of planted clones and selection rates among families, and vigor (7.2%) ...

CPCL 00-4111' (Reg. No. CV-144, PI 660673) sugarcane (a complex hybrid of Saccharum spp.) is the latest in a series of commercial sugarcane cultivar releases developed by the United States Sugar Corporation (USSC) and completed by the cooperative Canal Point sugarcane breeding and selection program, which included the USDA-ARS, the University of Florida, and the Florida Sugar Cane League, Inc. CPCL 00-4111, which was released in Florida on 18 Feb. 2011, was selected from a cross of genotype CL 83-3431 × CL 89-5189 made at Clewiston, FL on 4 Dec. 1998. The female and male parents, CL 83-3431 and CL 89-5189, respectively, are proprietary genotypes of USSC. CPCL 00-4111 was released because of its high cane yield and commercial recoverable sucrose on organic (muck) soils and because of its resistance to brown rust (caused by Puccinia melanocephala H. & P. Sydow), orange rust (caused by Puccinia kuehnii E.J. Butler), smut (caused by Ustilago scitaminea H. & P. Sydow), and Sugarcane mosaic virus strain E (the causal agent of mosaic), as well as its moderate resistance to leaf scald (caused by Xanthomonas albilineans (Ashby) Dowson). All of these diseases are of economic importance to sugarcane production in Florida.

Objective: To examine if wound aetiology has an effect on the pain-relieving properties of an ibuprofen-releasing foam dressing, which was previously shown to reduce pain in wounds of various aetiologies, compared with local best practice (LBP). Method: This was a secondary analysis of data from a multicentre, randomised, parallel group trial of patients with painful exuding wounds of various aetiologies. Wound aetiology was determined at enrolment. Of 853 patients enrolled into the trial, 688 belonged to a wound aetiology subgroup that included > 25 patients and were included in the analysis reported here. Patients were randomised to a dressing containing 112.5mg of ibuprofen (ibuprofen foam) or to LBP for 5 days. Patients recorded pain relief and pain intensity daily. The main endpoint was the proportion of patients who, from day 1 to day 5, reported a summed pain relief score > 50% of the total maximum pain relief score (TOTPARD5>50%) and the corresponding number needed to treat (NNT) for each wound aetiology subgroup. Further analyses included the proportion of patients who, on a daily basis, reported pain relief > 50% of the maximum daily pain relief, the proportion of patients who, on day 5, experienced a reduction in pain intensity of > 50% of the maximum score (PIDD5>50%), and if PIDD5>50% was related to baseline pain intensity. Results: Patients were categorised by the following five wound types: arterial, venous, and mixed arterial-venous leg ulcers, vasculitis and traumatic ulcers. The ibuprofen foam dressing was associated with significantly greater pain relief than LBP in all different wound aetiology subgroups, whether chronic or traumatic (acute). Overall, TOTPARD5>50% was 55% in the ibuprofen foam group and 24% in the LBP group (p < 0.0001; NNT, 3.2). The pain intensity evaluations revealed similar results in favour of ibuprofen foam compared with LBP. No correlation was observed between PIDD5>50 and initial pain intensity. Conclusion: In this study, the ibuprofen foam dressing was shown to consistently relieve wound pain in exuding wounds of various aetiologies, irrespective of basal pain intensity. The data suggest that local pain relief by an ibuprofen foam dressing is possible in the most common, painful, exuding, chronic and acute/traumatic wounds and so is a safer alternative to systemic pain treatment. Conflicts of interest: of the ten authors involved in the preparation of this manuscript, seven declare that they have no conflicts of interest. one is a member of the Wound Advisory Forum for Coloplast A/S, Denmark. Two are employees of Coloplast A/S, Denmark.

Research to develop ‘CPCL 99–4455’ (Reg. No. CV-142; PI 659802) sugarcane (a complex hybrid of Saccharum spp.) was initiated by the United States Sugar Corporation (USSC) and completed cooperatively by the USDA-ARS, the University of Florida, and the Florida Sugarcane League, Inc. CPCL 99–4455 was released to growers in Florida on 15 Dec. 2009. CPCL 99–4455 was selected from a cross of the USSC proprietary genotype CL 90–4643 × ‘CP 84–1198’ made at Clewiston, FL on 22 Dec. 1998. The female parent, CL 90–4643, has high yields of cane and sucrose but it is not used commercially by the USSC because of susceptibility to Sugarcane mosaic virus strain E. The male parent, cultivar CP 84–1198, has moderately high commercial recoverable sucrose (CRS) and cane yields and has been grown commercially on about 1–5% of Florida’s sugarcane acreage since 1996. CPCL 99–4455 is moderately resistant to smut (caused by Ustilago scitaminea Sydow & P. Sydow), brown rust (caused by Puccinia melanocephala H. & P. Sydow), orange rust (caused by Puccinia kuehnii E.J. Butler), leaf scald (caused by Xanthomonas albilineans Ashby, Dowson), and Sugarcane mosaic virus strain E. CPCL 99–4455 is expected to be used mostly on organic (muck) soils in Florida, where it had high CRS and acceptable yields of cane in our tests.

Sugarcane (a complex Saccharum hybrid) yellow leaf is caused by the Sugarcane yellow leaf virus (SCYLV) and is endemic in many growing regions. Inadequate sources of SCYLV resistance limit conventional breeding for resistance. Although transgenics offer the potential for improving SCYLV resistance, previous studies have shown that agronomic performance may be negatively impacted in primary (T0) transgenic lines due to somaclonal variation or transgene position effects. These problems could be overcome or reduced by transgene transfer to sexual progeny. The objectives of this study were to evaluate the utility as parents of two sugarcane genotypes (6-1 and 6-2) previously transformed with the nptII selectable marker and U-SCYLV-CP inserts and to examine the potential of kanamycin sensitivity as a selectable marker in progeny derived from these crosses. 6-1 and 6-2 were used as parents in 31 crosses in four crossing seasons. Flowering characteristics and germination rates of the resulting true seed were comparable with those for the untransformed parent, CP 92-1666, and with other crosses made in the same seasons. Inheritance and function of the nptII insert were tested in 215 seedlings from three crosses through the application of a solution containing 3.0 g L−1 kanamycin. Diagnostic PCR confirmed the presence of the nptII insert transgene in 158 (98.1%) of the 161 kanamycin-resistant progeny. Segregation analysis of the data suggests that the nptII insert is independently integrated at two linked loci in 6-1 one of which carries a functional copy of the nptII insert and the U-SCYLV-CP insert. Whereas for 6-2, nptII is inserted at a single, functional loci carrying the U-SCYLV-CP insert. ΔΔCt values were determined from real-time PCR data of each insert among progeny testing positive for both inserts using diagnostic PCR. This novel application of the ΔΔCt method allowed the determination of copy number relative to the T0 clone without the use of an exogenous control and showed that all copies of the U-SCYLV-CP insert were inherited among almost all progeny of either 6-1 or 6-2. These results demonstrate the utility of T0 transformed sugarcane genotypes for transmission of transgenes to additional sugarcane genotypes and the effectiveness of kanamycin application for screening sexual progeny and will be useful in guiding breeding strategies to improve resistance to the SCYLV through transgenics.

The phylogenetic positions of Puccinia spp. infecting sugarcane (a complex hybrid of Saccharum spp.) were determined using 38 newly generated rust sequences and 26 sequences from GenBank. Rust specimens on sugarcane were collected from 164 locations in 23 countries and identified based on light microscopy. The morphology for all samples matched that of Puccinia kuehnii or P. melanocephala, the orange and brown rust pathogens of sugarcane, respectively. Nuclear ribosomal DNA sequences (rDNA) including portions of the 5.8S rDNA, the complete internal transcribed spacer 2 (ITS2) and 5′ region of the large subunit (nLSU) rDNA were obtained for each species along with 36 additional rust taxa. Despite a shared host, the two Puccinia spp. on sugarcane are not closely related within the Pucciniales. Phylogenetic analyses place P. melanocephala most closely to P. miscanthi, P. nakanishikii, and P. rufipes infecting Miscanthus sinensis, Cymbopogon citratus, and Imperata cylindrica, respectively. Puccinia kuehnii is basal to a clade of Poaceae-infecting rusts including P. agrophila, P. polysora, P. substriata, and Uromyces setariae-italicae infecting Schizachyrium spp., Zea mays, Digitaria spp., and Urochloa mosambicensis, respectively. Light and scanning electron microscopy images highlight morphological differences distinguishing the two sugarcane-infecting species. This study confirms the separation of rust species infecting Poaceae from Cyperaceae- and Juncaceae-infecting rusts and also provides support for the presence of an additional group that includes P. kuehnii and other grass-infecting relatives.

Puccinia kuehnii and P. melanocephala cause orange and brown rust of sugarcane, respectively. Puccinia kuehnii has been confirmed in Asia, Australia and recently, the Caribbean basin, whereas P. melanocephala is distributed among the majority of sugarcane growing regions. Differentiating these two economically significant pathogens visually is problematic and limited to material exhibiting mature disease symptoms or spores. Partial ITS1, ITS2 and complete 5·8S sequences were generated from P. kuehnii and P. melanocephala isolates from around the world. PCR primers and dual labelled hydrolysis probes were designed for each pathogen for use in real-time PCR and optimized using locked nucleic acids (LNA). The primers amplified DNA from their target pathogens and not from other species of Puccinia or fungal species isolated from sugarcane leaves. Optimized real-time PCR conditions allowed the detection of 0·19 pg of P. kuehnii or P. melanocephala genomic DNA and differentiated the pathogens on sugarcane leaves prior to observing typical symptoms in the field. Primer-introduced restriction analysis-PCR (PIRA-PCR) was used to detect a single nucleotide polymorphism (Pk ITS1 183A>G) in ITS1 of P. kuehnii. Allele 183A was observed in all samples, whereas 183G was detected in 52% of samples from Asia and Australia yet absent from all Caribbean basin samples. Long distance spore dispersal, dispersal through an intermediate location or improper movement of contaminated material could explain the introduction of P. kuehnii to the Western hemisphere. However, the current proliferation of the pathogen in the Americas is limited to isolates which contain only the 183A allele.

Aims: To evaluate competitive PCR assays for quantifying seed-borne Microdochium and Fusarium seedling blight pathogen DNA and to determine test and year repeatability and sources of variability. Methods and Results: Relationships between DNA and plate counts were significant for Fusarium and Microdochium seedling blight pathogens in 152 seed batches from 3 years. Coefficient of determinations, however, differed greatly (Fusarium; R2 = 0·25, P = 0·029, Microdochium; R2 = 0·73, P 0·90). Residual maximum likelihood analysis showed that the least and greatest variance components of the testing procedure were DNA extraction subsampling and PCR assay respectively. Conclusions: Amount of pathogen DNA is a useful estimator of seed batch contamination for Microdochium but not Fusarium seedling blight pathogens. Although reproducible over time, improvements to the testing procedure should focus on repeated PCR amplifications to reduce assay variability. Significance and Impact of the Study: Replacing plate counts with competitive PCR for determining the severity of seed batch contamination is feasible in areas where Microdochium seedling blight pathogens predominate.

Improved yield and disease resistance on sand soils are priorities of the Canal Point (CP) sugarcane (Saccharum spp.) breeding and selection program. Analyses of historical phenotypic data can provide helpful information in guiding selection strategies to meet these priorities. Correlation analysis was used to examine repeatability of phenotypic data used to advance genotypes from an unreplicated single location clonal crop test (stage II) to the subsequent stage (stage III; two replicate, four location clonal crop experiment). Correlations between data for four traits measured in stage II and the corresponding data pooled across soil types for the same genotypes in stage III varied across 23 series of the CP program. Generally, when correlations were statistically significant (P < 0.05), correlation values were low (means; theoretical recoverable sucrose (TRS) r = 0.40, cane yield r = 0.27, and economic index r = 0.23). Similar trends were evident for correlations between data from stage II and stage III...

Sugarcane is a proven biofuel feedstock and accounts for about 40% of the biofuel production worldwide. It has a more favorable energy input/output ratio than that of corn, the other major biofuel feedstock. The rich resource of genetic diversity and the plasticity of autopolyploid genomes offer a wealth of opportunities for the application of genomics and technologies to address fundamental questions in sugarcane towards maximizing biomass production. In a workshop on sugarcane engineering held at Rutgers University, we identified research areas and emerging technologies that could have significant impact on sugarcane improvement. Traditional plant physiological studies and standardized phenotypic characterization of sugarcane are essential for dissecting the developmental processes and patterns of gene expression in this complex polyploid species. Breeder friendly DNA markers associated with target traits will enhance selection efficiency and shorten the long breeding cycles. Integration of cold tolerance from Saccharum spontaneum and Miscanthus has the potential to expand the geographical range of sugarcane production from tropical and subtropical regions to temperate zones. The Flex-stock and mix-stock concepts could be solutions for sustaining local biorefineries where no single biofuel feedstock could provide consistent year-round supplies. The ever increasing capacities of genomics and biotechnologies pave the way for fully exploring these potentials to optimize sugarcane for biofuel production.

Research to develop ‘CPCL 97-2730’ (Reg. No. CV-137, PI 655943) sugarcane (a complex hybrid of Saccharum spp.) was initiated by the United States Sugar Corporation (USSC) and completed cooperatively by the USDA–ARS, the University of Florida, and the Florida Sugar Cane League, Inc. CPCL 97-2730 was released to growers in Florida in December 2008. CPCL 97-2730 was selected from a cross of USSC proprietary genotypes CL 75-0853 × CL 88-4730 made at Clewiston, FL, in December 1996. The female parent, CL 75-0853, had high commercial recoverable sucrose (CRS) but because of low cane yields was not used extensively by USSC. The male parent, CL 88-4730, is a high-yielding genotype that USSC is evaluating in commercial fi elds with sand and muck soils for possible extended use. CPCL 97-2730 was released because of its acceptable yields of cane and CRS on sand soils and its resistance to smut (caused by Ustilago scitaminea Syd. & P. Syd.), brown rust (caused by Puccinia melanocephala Syd. & P. Syd.), orange rust [caused by P. kuehnii (Kruger) E. Butler], leaf scald [caused by Xanthomonas albilineans (Ashby) Dowson], and sugarcane mosaic virus strain E.

'CP 01-1372' (Reg. No. CV-136, PI 655942) sugarcane (a complex hybrid of Saccharum spp.) was developed through cooperative research conducted by the USDA―ARS, the University of Florida, and the Florida Sugar Cane League, Inc., and was released to growers in Florida in December 2008. CP 01-1372 was selected from a cross of genotype CP 94-1200 x 'CP 89-2143' made at Canal Point, FL, in December 1998. The female parent, CP 94-1200, was advanced to the final selection stage (Stage 4) of the Canal Point sugarcane cultivar breeding and selection program. The male parent, CP 89-2143, is a major sugarcane cultivar in Florida. CP 01-1372 was released because of its high cane yield and commercial recoverable sucrose on both organic and sand soils, and its acceptable levels of resistance to brown rust (caused by Puccinia melanocephala Syd. & P. Syd.), orange rust [caused by P. kuehnii (Kruger) E. Butler], leaf scald [caused by Xanthomonas albilineans (Ashby) Dowson], Sugarcane mosaic virus strain E (mosaic), and ratoon stunt (caused by Clavibacterxyli subsp. xyli Davis et al.) in Florida.

Sugarcane yellow leaf virus (SCYLV, a Poleovirus of the Luteoviridae family) is already widespread in Florida, and resistance in the Canal Point (CP) sugarcane population is limited. Genetic transformation of sugarcane for disease resistance holds promise but tissue culture and transformation processes produce undesirable agronomic characteristics necessitating thorough field evaluation. A 3-year sugarcane (a complex hybrid of Saccharum species) field study was conducted in Belle Glade, FL with the following objectives: (1) thoroughly evaluate the agronomic performance of two transgenic lines transformed for SCYLV resistance (6-1, 6-2) compared with parental cultivar control CP 92-1666, (2) determine level of SCYLV resistance in the transgenic lines, and (3) characterize genetic differences in the transgenic lines compared with CP 92-1666 using simple sequence repeat (SSR) genotyping. Sugarcane yields of CP 92-1666 were superior to both transgenic lines, as well as tissue culture (C-1) and nptII marker gene (20-1) controls, in the plant cane, first ratoon and second ratoon crops. CP 92-1666 recorded an average of 6.5–8.7 tons sucrose ha−1 yr−1 more than genotypes subjected to tissue culture and biolistic transformation. However, SCYLV infection rates in transgenic lines were only 0–5%, compared with 98% in CP 92-1666. Kanamycin field assays indicated that selectable marker gene nptII was stably expressed in all co-transformed lines. SSR genotyping showed 35 additional fragments to be present and 25 existing fragments absent among 6-1, 6-2, C-1 and 20-1 compared with CP 92-1666. Although all clones had unique genotypes, the four regenerated clones showed a greater genetic distance from the donor clone CP 92-1666 (mean GD 0.4) than to one another (mean GD 0.03). This study reports the first successful gene transfer of SCYLV resistance in sugarcane and the first report of variations in microsatellite repeat number associated with regeneration from embryogenic callus. Our results highlight the potential of genetic transformation methods to incorporate desirable traits into sugarcane, combined with the necessity of thorough agronomic evaluation of transgenic genotypes. Transgenic lines 6-1 and 6-2 are being used as parents in crosses designed to combine SCYLV resistance from these genotypes with agronomic characteristics of high-yielding materials.

CP 00-1446' (Reg. No CV-133, PI 654092) sugarcane (a complex hybrid of Saccharum spp.) was developed through cooperative research conducted by the USDA-ARS, the University of Florida, and the Florida Sugar Cane League, Inc., and was released to growers in Florida in September 2007. CP 00-1446 was selected from a cross of genotypes CP 93-1607 × CP 91-1150 made at Canal Point, FL, in January 1998. The female and male parents were advanced to the penultimate selection stage (Stage 3) and the fi nal stage (Stage 4), respectively, of the Canal Point sugarcane cultivar breeding and selection program. CP 00-1446 was released and recommended for sand soils in Florida because of its high plant cane and acceptable ratoon per hectare yields of cane and sucrose and commercial recoverable sucrose on sand soils, and its acceptable disease reactions to smut (caused by Ustilago scitaminea (Sydow & P. Sydow)) (moderately susceptible), brown rust (caused by Puccinia melanocephala H. & P. Sydow) (moderately resistant), orange rust (caused by Puccinia kuehnii E.J. Butler) (moderately resistant), leaf scald (caused by Xanthomonas albilineans Ashby, Dowson) (resistant), Sugarcane mosaic virus strain E (mosaic) (moderately susceptible), and ratoon stunting disease (caused by Clavibacter xyli subsp. xyli Davis) (resistant) in Florida.

'CP 00-2180' (Reg. No. CV-134, PI 654093) sugarcane (a complex hybrid of Saccharum spp.) was developed through cooperative research conducted by the USDA-ARS, the University of Florida, and the Florida Sugar Cane League, Inc., and was released to growers in Florida in September 2007. CP 00-2180 was selected from a self-cross of cultivar HoCP 91-552 made at Canal Point, FL, in January 1998. Based on its high cane yields and fiber content (16%), HoCP 91-552 was released as a cultivar for bioenergy in Louisiana. CP 00-2180 was released and recommended for sand soils in Florida because of its high plant cane and acceptable ratoon per hectare yields of cane and sucrose and commercial recoverable sucrose on sand soils, and its resistance to smut [caused by Ustilago scitaminea (Sydow & P. Sydow)], brown rust (caused by Puccinia melanocephala H. & P. Sydow), orange rust (caused by Puccinia kuehnii E.J. Butler), leaf scald [caused by Xanthomonas albilineans (Ashby) Dowson], Sugarcane mosaic virus strain E, and ratoon stunting disease (caused by Clavibacter xyli subsp. xyli Davis) in Florida.

Breast augmentation and breast reconstruction, either immediate or delayed, are increasingly common operations. All radiologists need to be able to recognize the normal appearances of the more commonly used implants on various imaging modalities, and breast radiologists in particular are facing new challenges when imaging the women involved. This article aims to review the normal and abnormal findings in women who have had breast augmentation and reconstruction, including implant insertion and reconstruction by autologous myocutaneous flaps.

'CP 88-1165' (Reg. No. CV-131, PI 651884) sugarcane (a complex hybrid of Saccharum spp.) was developed through cooperative research conducted by the USDA-ARS, the Centro Guatemalteco de Investigacion y Capacitacion de la Cana de Azucar, and the University of Florida. It was released to growers in Guatemala in September 2004. CP 88-1165 was selected from the cross 'CL 61-620' x CP 81-1302 made at Canal Point, FL, in January 1986. The female parent, CL 61-620, formerly a major commercial cultivar in Florida, was developed by a private breeding program in Clewiston, FL, conducted by the United States Sugar Corp. The male parent, CP 81-1302, was advanced to the final testing stage (Stage 4) of the Canal Point (CP) cultivar development program but was not released due to susceptibility to brown rust (caused by Puccinia melanocephala Syd. & P. Syd.). CP 88-1165 was not released in Florida because of its susceptibility to brown rust and low yield of commercial recoverable sucrose. However, after testing in Guatemala, CP 88-1165 was released there because of its high plant cane and ratoon per hectare yields of cane and sucrose and its acceptable resistance or tolerance to smut [caused by Ustilago scitaminea (Sydow & P. Sydow], brown rust, Sugarcane mosaic virus strain E (mosaic), Sugarcane yellow leaf virus, and red stripe (Acidovorax avenae subsp. avenae) in Guatemala.

BACKGROUND: Resistance gene analogues (RGAs) have been isolated from many crops and offer potential in breeding for disease resistance through marker-assisted selection, either as closely linked or as perfect markers. Many R-gene sequences contain kinase domains, and indeed kinase genes have been reported as being proximal to R-genes, making kinase analogues an additionally promising target. The first step towards utilizing RGAs as markers for disease resistance is isolation and characterization of the sequences. RESULTS: Sugarcane clone US01-1158 was identified as resistant to yellow leaf caused by the sugarcane yellow leaf virus (SCYLV) and moderately resistant to rust caused by Puccinia melanocephala Sydow & Sydow. Degenerate primers that had previously proved useful for isolating RGAs and kinase analogues in wheat and soybean were used to amplify DNA from sugarcane (Saccharum spp.) clone US-01-1158. Sequences generated from 1512 positive clones were assembled into 134 contigs of between two and 105 sequences. Comparison of the contig consensuses with the NCBI sequence database using BLASTx showed that 20 had sequence homology to nuclear binding site and leucine rich repeat (NBS-LRR) RGAs, and eight to kinase genes. Alignment of the deduced amino acid sequences with similar sequences from the NCBI database allowed the identification of several conserved domains. The alignment and resulting phenetic tree showed that many of the sequences had greater similarity to sequences from other species than to one another. CONCLUSION: The use of degenerate primers is a useful method for isolating novel sugarcane RGA and kinase gene analogues. Further studies are needed to evaluate the role of these genes in disease resistance. Published in 2007 by John Wiley & Sons, Ltd.

Degenerate PCR primers were designed based on EF-1 alpha (EF-1alpha) gene sequences of several filamentous fungi retrieved from sequence databases. These primers were used to isolate a partial sequence, approximately 830 bp in length of the EF-1alpha from isolates of Microdochium nivale obtained from various geographic locations across the world. Two distinct groups of isolates were evident among those isolates examined. Sequence homology for comparisons within group was 99.7% for group A and 99.8% for group B. Primers specific to either group A or group B sequences were designed and tested on isolates from around the world. Comparisons were made with primers previously reported for the two varieties of M. nivale and revealed that Group A type isolates correlated with M. nivale var. majus and group B isolates with M. nivale var. nivale. The primers from this study and those previously reported were in agreement for all isolates with the exception of one isolate (NRRL 3289) which failed to amplify with previously published M. nivale primers. Sequence analysis of NRRL 3289 suggested that it was an isolate of M. nivale var. nivale as indicated by the EF-1alpha based primers developed in this study. This study provides sequence based phylogenetic evidence of two species and when taken together with biological differences reported, leads to the recognition of M. majus comb. nov. (syn. Fusarium nivale var. majus). Descriptions of the two species are provided.

Sugarcane (Saccharum spp.) and other members of Saccharum spp. are attractive biofuel feedstocks. One of the two World Collections of Sugarcane and Related Grasses (WCSRG) is in Miami, FL. This WCSRG has 1002 accessions, presumably with valuable alleles for biomass, other important agronomic traits, and stress resistance. However, the WCSRG has not been fully exploited by breeders due to its lack of characterization and unmanageable population. In order to optimize the use of this genetic resource, we aim to 1) genotypically evaluate all the 1002 accessions to understand its genetic diversity and population structure and 2) form a core collection, which captures most of the genetic diversity in the WCSRG. We screened 36 microsatellite markers on 1002 genotypes and recorded 209 alleles. Genetic diversity of the WCSRG ranged from 0 to 0.5 with an average of 0.304. The population structure analysis and principal coordinate analysis revealed three clusters with all S. spontaneum in one cluster, S. officinarum and S. hybrids in the second cluster and mostly non-Saccharum spp. in the third cluster. A core collection of 300 accessions was identified which captured the maximum genetic diversity of the entire WCSRG which can be further exploited for sugarcane and energy cane breeding. Sugarcane and energy cane breeders can effectively utilize this core collection for cultivar improvement. Further, the core collection can provide resources for forming an association panel to evaluate the traits of agronomic and commercial importance.

Background Accuracy of predictive formulae is crucial for therapeutic planning because indirect calorimetry measurement is not always possible or cost effective. Energy requirements are more difficult to predict in the acutely ill obese patient compared with lean patients because of an increased resting energy expenditure per lean body mass and a variable stress response to illness. Methods A retrospective review of 726 patients identified 57 patients (32 spontaneous breathing, S; 25 ventilator dependent, V) with body mass indexes of 30-50 kg/m2. Limits-of-agreement analysis determined bias (the mean difference between measured and predicted values) and precision (the standard deviation of the bias) to evaluate the accuracy of predictive formulae compared with measured resting energy expenditure (MREE) by a Deltatrac Metabolic Monitor. Predictive accuracy was determined within+/-10% MREE. The predictive formulae examined were: variations of the Harris-Benedict equations using ideal, adjusted weights of 25% and 50% and actual weights with stress factors ranging from 1.0 to 1.5; the Ireton-Jones equation for obesity; the Ireton-Jones equations for hospitalized patients (S and V); and the ratio of 21 kcalories per kilogram actual weight. Results The mean MREE was 21 kcal/kg actual weight. The adjusted Harris-Benedict average weight equation was optimal for predicting MREE for the combined S and V sets (bias = 182+/-123; 67%+/-10% MREE), as well as the S subset (bias = 159+/-112; 69%+/-10% MREE). Conclusions The Harris-Benedict equations using the average of actual and ideal weight and a stress factor of 1.3 most accurately predicted MREE in acutely ill, obese patients with BMIs of 30-50 kg/m2. Predictive formulae were least accurate for obese, ventilator-dependent patients.