Immediate impacts of soybean cover crop on bacterial community composition and diversity in soil under long-term Saccharum monoculture.

Saccharum yield decline results from long-term monoculture practices. Changes in cropping management can improve soil health and productivity. Below-ground bacterial community diversity and composition across soybean (Glycine max (L.) Merr) cover crop, Saccharum monoculture (30+ year) and fallowed soil were determined. Near full length (~1,400 base pairs) of 16S rRNA gene sequences were extracted from the rhizospheres of sugarcane and soybean and fallowed soil were compared. Higher soil bacterial diversity was observed in the soybean cover crop than sugarcane monoculture across all measured indices (observed operationational taxonomic units, Chao1, Shannon, reciprocal Simpson and Jackknife). Acidocateria, Proteobacteria, Bacteroidetes and Planctomycetes were the most abundant bacterial phyla across the treatments. Indicator species analysis identified nine indicator phyla. Planctomycetes, Armatimonadetes and candidate phylum FBP were associated with soybean; Proteobacteria and Firmicutes were linked with sugarcane and Gemmatimonadetes, Nitrospirae, Rokubacteria and unclassified bacteria were associated with fallowed soil. Non-metric multidimensional scaling analysis showed distinct groupings of bacterial operational taxonomic units (97% identity) according to management system (soybean, sugarcane or fallow) indicating compositional differences among treatments. This is confirmed by the results of the multi-response permutation procedures (A = 0.541, p = 0.00045716). No correlation between soil parameters and bacterial community structure was observed according to Mantel test (r = 211865, p = 0.14). Use of soybean cover-crop fostered bacterial diversity and altered community structure. This indicates cover crops could have a restorative effect and potentially promote sustainability in long-term Saccharum production systems.

An enriched sugarcane diversity panel for utilization in genetic improvement of sugarcane.

Sugarcane crop is important for both sugar and biofuels. A world collection of sugarcane and related grasses (WCSRG) maintained at Miami, FL contains > 1,200 non-redundant clones of different species and genera within the Saccharum complex. However, linkage of undesirable alleles with useful genes in wild species has hindered its efficient utilization in sugarcane breeding. A core collection developed previously with smaller number of clones representing WCSRG did not take into account > 120 wild/exotic clones maintained at the USDA-ARS Sugarcane Research Unit in Houma, Louisiana. Moreover, the genome complexity and sub-tropical to temperate growing climate of Louisiana warrant a region-specific core collection that can be used for base-broadening breeding aimed at efficient introgression of desirable alleles. Genetic diversity of 1,485 clones within WCSRG and Louisiana (commercials, wild/exotic) using 423 SSR alleles showed an average gene diversity (h) at 0.208 among all species groups where Erianthus-like Saccharum species (ELSS), Miscanthus spp., and S. spontaneum each formed a distinct cluster, Saccharum robustum, S. officinarum, hybrid cultivars, and S. edule grouped together in a major cluster, and Saccharum sinense and S. barberi formed distinct grouping. A 309-clone diversity panel (SDP1) was developed that captured the genetic diversity based on the combination of maximum length subtree and manual selection to maximize representation of Louisiana clones and minimize import of clones from Miami. SDP1 shared 324 alleles out of the 423 alleles in the entire population of 1,485 clones and captured the genetic diversity of the entire collection with an average gene diversity (h) at 0.163. The variation within (11-17%) and among (83-89%) the populations in SDP1 were comparable with the entire population of 1,485 clones (9-15% and 85-91%, respectively). The breadth of the genetic variation of SDP1 was exemplified by the intra- and inter-specific diversity of a 190-clone mini-core collection with markers derived from known cold-responsive genes. SDP1 will facilitate genome-wide association studies for identification of trait-specific markers for use in marker-assisted breeding in Louisiana and elsewhere.

Sugarcane cultivar response to glyphosate and trinexapac-ethyl ripeners in Louisiana.

Sugarcane ripening in Louisiana is necessary to ensure adequate sucrose levels in early-season harvested sugarcane. The response of nine sugarcane cultivar's yield components to glyphosate and trinexapac-ethyl ripeners was determined in field trials. Glyphosate (210 g ae ha-1) and trinexapac-ethyl (200 g ai ha-1) treatments failed to increase sucrose yields more than non-ripened sugarcane. Sugarcane ripened with glyphosate or trinexapac-ethyl increased theoretical recoverable sucrose (TRS) 4 to 12% more than non-ripened sugarcane in seven out of nine cultivars, but greater TRS values were counterpoised by lower sugarcane stalk weight. An unintentional consequence of reduced late-season vegetative growth may benefit growers by allowing them to harvest more sugarcane hectares to meet their daily load quota and exposes fewer hectares to a freeze event. The cultivars HoCP 00-950, Ho 09-804, and HoCP 09-840 were not responsive to glyphosate or trinexapac-ethyl ripeners and should not be treated. A delayed harvest from 28 to 49 days after treatment (DAT) coincided with greater TRS values and 17% more sucrose yield.

Genome-wide association mapping identifies markers associated with cane yield components and sucrose traits in the Louisiana sugarcane core collection.

Sugarcane is an economically important crop for both food and biofuel industries. Marker-assisted breeding in sugarcane is becoming a reality with the recent development and deployment of markers linked with disease resistance genes. Large linkage disequilibrium in sugarcane makes genome-wide association studies (GWAS) a better alternative to biparental mapping to identify markers associated with agronomic traits. GWAS was conducted on a Louisiana core collection to identify marker-trait associations (MTA) for 11 cane yield and sucrose traits using single nucleotide polymorphism (SNP) and insertion-deletion (Indel) markers. Significant (P < .05) MTAs were identified for all traits where the top ranked markers explained up to 15% of the total phenotypic variation. High correlations (0.732 to 0.999) were observed between sucrose traits and 56 markers were found consistent across multiple traits. These markers following validation in more diverse populations could be used in marker-assisted selection of clones in sugarcane breeding program in Louisiana and elsewhere.

Effect of growing media and fertilization on sugarcane flowering under artificial photoperiod.

The USDA-ARS Sugarcane Variety Development Program in Houma, LA aims to maximize the number of panicles available for crossing through artificial manipulation of the environment. In a three-year study, the effect of growing media, fertilizer treatment, and their interaction on sugarcane flowering (% of panicles emerged), and number of days to flowering (DTF) under an artificial photoperiod treatment were assessed. The commercially-available sugarcane cultivar, 'HoCP 96-540' was planted in 2.8-L pots and subjected to the standard local photoperiod treatment. The cultivar was planted in four growing media (RediEarth Seedling and Germination Mix, Fafard, Metro-Mix®902, and Metro-Mix®900) and subjected to three different fertilizer applications. In the control treatment, fertilizer application was stopped prior to the commencement of the photoperiod treatment as practiced in some sugarcane breeding programs. The continuous treatment consisted of an application of a 10 ml solution of a NPK three times a week between June and October. The partial treatment consisted of applications of the same NPK solution applied post-initiation between September and October. Nitrogen starvation prior to the commencement of the photoperiod treatment is generally accepted to improve flower initiation; thus the standard practice is to cease nitrogen application two weeks prior to beginning a photoperiod regime. The growing media used in this study did not have a significant effect on days to flowering or percent panicle emergence. In our study, the control fertilizer treatment showed a flowering percentage across all growing media types of 21.2% less than a continuous fertilization regime. Furthermore, a significant trend was observed between fertilization treatments and days to flowering, with the continuous treatment producing panicles, on average across growing media, four days earlier than the control treatment, and six days earlier than the partial treatment. Evidence across this three-year experiment indicates that we should consider modifying plant nutrition management as soil fertility was found to be inadequate.

Estimating Broad Sense Heritability and Investigating the Mechanism of Genetic Transmission of Cold Tolerance Using Mannitol as a Measure of Post-freeze Juice Degradation in Sugarcane and Energycane (Saccharum spp.).

In approximately 25% of the sugarcane-producing countries worldwide, conventional sugarcane (Saccharum spp. hybrids) is exposed to damaging freezes. A study was conducted during the 2009 and 2010 harvest seasons to compare late-season freeze tolerance among three groups: commercial Louisiana sugarcane genotypes, early generation genotypes selected for cold tolerance in the U.S. Department of Agriculture sugarcane breeding programs at Houma, LA, and Canal Point, FL, and potential energycane genotypes selected for high total biomass per acre. Mannitol concentrations in cane juice following freezing temperatures were determined to evaluate levels of cold tolerance. Genotypes selected for cold tolerance in Houma, LA, had significantly more late-season freeze tolerance than commercial sugarcane genotypes and genotypes selected in Canal Point, FL. Genotypes showing the most cold tolerance were Ho02-146 and Ho02-152, and those that were most highly susceptible were US87-1006 and US87-1003 (early-generation breeding genotypes) and L99-233 (commercial genotype). Broad-sense heritability for late-season cold tolerance in the two-year study was estimated at g(2) = 0.78. The enzymatic mannitol analysis successfully differentiated high-fiber energycane genotypes from those from other sources.

Identification of cold-responsive genes in energycane for their use in genetic diversity analysis and future functional marker development.

Breeding for cold tolerance in sugarcane will allow its cultivation as a dedicated biomass crop in cold environments. Development of functional markers to facilitate marker-assisted breeding requires identification of cold stress tolerance genes. Using suppression subtractive hybridization, 465 cold-responsive genes were isolated from the cold-tolerant energycane Ho02-144. Predicted gene interactions network indicated several associated pathways that may coordinately regulate cold tolerance responses in energycane. Expression analysis of a select set of genes, representing signaling and transcription factors, genes involved in polyamine and antioxidant biosynthesis, protein degradation and in the repair of damaged proteins in the cytosol, showed their time-dependent regulation under cold-stress. Comparative expression profiles of these genes between Ho02-144 and a cold-sensitive clone (L79-1002) showed that almost all genes were induced immediately upon imposition of cold stress and maintained their expression in Ho02-144 whereas they were either downregulated or their upregulation was very low in L79-1002. Simple sequence repeat markers derived from 260 cold-responsive genes showed allelic diversity among the cold-sensitive commercial hybrids that were distinct from the Saccharum spontaneum clones. Future efforts will target sequence polymorphism information of these genes in our ongoing QTL and association mapping studies to identify functional markers associated with cold tolerance in sugar/energycane.

Genotype, location, and year influence antioxidant activity, carotenoid content, phenolic content, and composition in specialty potatoes.

The influence of genotype, location, and year on antioxidant activity (AOA), total phenolics (TP), total carotenoids (TC), and phenolic composition was studied using specialty (colored) potatoes ( Solanum tuberosum L.) from the Texas Potato Variety Development Program. Twenty-five potato genotypes were grown at two Texas locations (McCook and Dalhart) and in two years (2003 and 2004). The AOA, TP, and TC differed significantly with genotype (G), location (L), and year (Y). Phenolic composition differed significantly among genotypes and between locations. The AOA, TP, and chlorogenic acid content were significantly correlated with one another. Genotypic effects were significant for all parameters measured and were larger than location and year effects. Interaction effects (G x L and G x L x Y) were significant for most parameters, but were relatively smaller than genotypic effects. Lutein and violaxanthin were the major carotenoids identified, and genotypes differed significantly in their carotenoid content. Genotypes CO112F2-2P/P and ATTX98013-1R/R were stable between locations and years with high AOA and TP, suggesting that they could be used as parents in breeding varieties with improved health benefits.

Heterosis for horticultural traits in broccoli.

Over the last three decades, broccoli (Brassica oleracea L., Italica Group) hybrids made by crossing two inbred lines replaced open-pollinated populations to become the predominant type of cultivar. The change to hybrids evolved with little or no understanding of heterosis or hybrid vigor in this crop. Therefore, the purpose of the present study was to determine levels of heterosis expressed by a set of hybrids derived by crossing relatively elite, modern inbreds (n = 9). An additional objective was to determine if PCR-based marker derived genetic similarities among the parents can be useful to predict heterosis in this crop. Thirty-six hybrids derived from a diallel mating design involving nine parents were evaluated for five horticultural characters including the head characteristics of head weight, head stem diameter, and maturity (e.g., days from transplant to harvest), and the plant vigor characteristics of plant height, and plant width in four environments. A total of 409 polymorphic markers were generated by 24 AFLP, 23 SRAP and 17 SSR primer combinations. Euclidean distances between parents were determined based on phenotypic traits. About half of the hybrids exhibited highparent heterosis for head weight (1-30 g) and stem diameter (0.2-3.5 cm) when averaged across environments. Almost all hybrids showed highparent heterosis for plant height (1-10 cm) and width (2-13 cm). Unlike other traits, there was negative heterosis for maturity, indicating that heterosis for this character in hybrids is expressed as earliness. Genetic similarity estimates among the nine parental lines ranged from 0.43 to 0.71 and were significantly and negatively correlated with highparent heterosis for all traits except for stem diameter and days from transplant to harvest. Euclidean distances were not correlated with heterosis. With modern broccoli inbreds, less heterosis was observed for head characteristics than for traits that measured plant vigor. In addition, genetic similarity based on molecular markers was more highly correlated with plant vigor characteristics than head traits. Unlike with molecular marker-based estimates of genetic similarity, euclidean distance determined using phenotypic trait data was not predictive of heterosis. In conclusion, this study has documented heterosis in Brassica oleracea L., and the ability to predict heterosis in this crop using molecular marker-based estimates of genetic similarity among parents used in producing the hybrid.