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Why Black Flowers? An Extreme Environment and Molecular Perspective of Black Color Accumulation in the Ornamental and Food Crops

Pollinators are attracted to vibrant flower colors. That is why flower color is the key agent to allow successful fruit set in food or ornamental crops. However, black flower color is the least attractive to pollinators, although a number of plant species produce black flowers. Cyanidin-based anthocyanins are thought to be the key agents to induce black color in the ornamental and fruit crops. R2R3-MYB transcription factors (TFs) play key roles for the tissue-specific accumulation of anthocyanin. MYB1 and MYB11 are the key TFs regulating the expression of anthocyanin biosynthesis genes for black color accumulation. Post-transcriptional silencing of flavone synthase II ( FNS ) gene is the technological method to stimulate the accumulation of cyanidin-based anthocyanins in black cultivars. Type 1 promoter of DvIVS takes the advantage of FNS silencing to produce large amounts of black anthocyanins. Exogenous ethylene application triggers anthocyanin accumulation in the fruit skin at ripening. Environment cues have been the pivotal regulators to allow differential accumulation of anthocyanins to regulate black color. Heat stress is one of the most important environmental stimulus that regulates concentration gradient of anthocyanins in various plant parts, thereby affecting the color pattern of flowers. Stability of black anthocyanins in the extreme environments can save the damage, especially in fruits, caused by abiotic stress. White flowers without anthocyanin face more damages from abiotic stress than dark color flowers. The intensity and pattern of flower color accumulation determine the overall fruit set, thereby controlling crop yield and human food needs. This review paper presents comprehensive knowledge of black flower regulation as affected by high temperature stress, and the molecular regulators of anthocyanin for black color in ornamental and food crops. It also discusses the black color-pollination interaction pattern affected by heat stress for food and ornamental crops.

Ahmad, Sagheer↗

Protoplast-Based Transient Expression and Gene Editing in Shrub Willow ( Salix purpurea L .)

Shrub willows (Salix section Vetrix) are grown as a bioenergy crop in multiple countries and as ornamentals across the northern hemisphere. To facilitate the breeding and genetic advancement of shrub willow, there is a strong interest in the characterization and functional validation of genes involved in plant growth and biomass production. While protocols for shoot regeneration in tissue culture and production of stably transformed lines have greatly advanced this research in the closely related genus Populus, a lack of efficient methods for regeneration and transformation has stymied similar advancements in willow functional genomics. Moreover, transient expression assays in willow have been limited to callus tissue and hairy root systems. Here we report an efficient method for protoplast isolation from S. purpurea leaf tissue, along with transient overexpression and CRISPR-Cas9 mediated mutations. This is the first such report of transient gene expression in Salix protoplasts as well as the first application of CRISPR technology in this genus. These new capabilities pave the way for future functional genomics studies in this important bioenergy and ornamental crop.

59 BASIC BIOLOGICAL SCIENCES↗

Taxonomy and Identification of Principal Foliar Nematode Species ( Aphelenchoides and Litylenchus )

Nematodes are Earth’s most numerous multicellular animals and include species that feed on bacteria, fungi, plants, insects, and animals. Foliar nematodes are mostly pathogens of ornamental crops in greenhouses, nurseries, forest trees, and field crops. Nematode identification has traditionally relied on morphological and anatomical characters using light microscopy and, in some cases, scanning electron microscopy (SEM). This review focuses on morphometrical and brief molecular details and key characteristics of some of the most widely distributed and economically important foliar nematodes that can aid in their identification. Aphelenchoides genus includes some of the most widely distributed nematodes that can cause crop damages and losses to agricultural, horticultural, and forestry crops. Morphological details of the most common species of Aphelenchoides (A. besseyi, A. bicaudatus, A. fragariae, A. ritzemabosi) are given with brief molecular details, including distribution, identification, conclusion, and future directions, as well as an updated list of the nominal species with its synonyms. Litylenchus is a relatively new genus described in 2011 and includes two species and one subspecies. Species included in the Litylenchus are important emerging foliar pathogens parasitizing trees and bushes, especially beech trees in the United States of America. Brief morphological details of all Litylenchus species are provided.

54 ENVIRONMENTAL SCIENCES↗

Advanced Life Support Systems: Opportunities for Technology Transfer

NASA's future missions to explore the solar system will be of long-duration possibly lasting years at a time. Human life support systems will have to operate with very high reliability for these long periods with essentially no resupply from Earth. Such life support systems will make extensive use of higher plants, microorganisms, and physicochemical processes for recycling air and water, processing wastes, and producing food. Development of regenerative life support systems will be a pivotal capability for NASA's future human missions. A fully functional closed loop human life support system currently does not exist and thus represents a major technical challenge for space exploration. Technologies where all life support consumables are recycled have many potential terrestrial applications as well. Potential applications include providing human habitation in hostile environments such as the polar regions or the desert in such a way as to minimize energy expenditures and to minimize negative impacts on those often ecologically-sensitive areas. Other potential applications include production of food and ornamental crops without damaging the environment from fertilizers that contaminate water supplies; removal of trace gas contaminants from tightly sealed, energy-efficient buildings (the so-called sick building syndrome); and even the potential of gaining insight into the dynamics of the Earth's biosphere such that we can better manage our global environment. Two specific advanced life support technologies being developed by NASA, with potential terrestrial application, are the zeoponic plant growth system and the Hybrid Regenerative Water Recovery System (HRWRS). The potential applications for these candidate dual use technologies are quite different as are the mechanisms for transfer. In the case of zeoponics, a variety of commercial applications has been suggested which represent potentially lucrative markets. Also, the patented nature of this product offers opportunities for licensing to commercial entities. In the case of the HRWRS, commercial markets with broad applications have not been identified but some terrestrial applications are being explored where this approach has advantages over other methods of waste water processing. Although these potential applications do not appear to have the same broad attraction from the standpoint of rapid commercialization, they represent niches where commercialization possibilities as well as social benefits could be realized.

Fields, B.↗

Feasibility of Estimating Relative Nutrient Contributions of Agriculture using MODIS Time Series

Around the Gulf of Mexico, high-input crops in several regions make a significant contribution to nutrient loading of small to medium estuaries and to the near-shore Gulf. Some crops cultivated near the coast include sorghum in Texas, rice in Texas and Louisiana, sugarcane in Florida and Louisiana, citrus orchards in Florida, pecan orchards in Mississippi and Alabama, and heavy sod and ornamental production around Mobile and Tampa Bay. In addition to crops, management of timberlands in proximity to the coasts also plays a role in nutrient loading. In the summer of 2008, a feasibility project is planned to explore the use of NASA data to enhance the spatial and temporal resolution of near-coast nutrient source information available to the coastal community. The purpose of this project is to demonstrate the viability of nutrient source information products applicable to small to medium watersheds surrounding the Gulf of Mexico. Conceptually, these products are intended to complement estuarine nutrient monitoring.

Ross, Kenton W.↗

Feasibility of Estimating Relative Nutrient Contributions of Agriculture and Forests Using MODIS Time Series

Around the Gulf of Mexico, high-input crops in several regions make a significant contribution to nutrient loading of small to medium estuaries and to the near-shore Gulf. Some crops cultivated near the coast include sorghum in Texas, rice in Texas and Louisiana, sugarcane in Florida and Louisiana, citrus orchards in Florida, pecan orchards in Mississippi and Alabama, and heavy sod and ornamental production around Mobile and Tampa Bay. In addition to crops, management of timberlands in proximity to the coasts also plays a role in nutrient loading. In the summer of 2008, a feasibility project is planned to explore the use of NASA data to enhance the spatial and temporal resolution of near-coast nutrient source information available to the coastal community. The purpose of this project is to demonstrate the viability of nutrient source information products applicable to small to medium watersheds surrounding the Gulf of Mexico. Conceptually, these products are intended to complement estuarine nutrient monitoring.

Ross, Kenton W.↗

Systematic approaches to C-lignin engineering in Medicago truncatula

Abstract Background C-lignin is a homopolymer of caffeyl alcohol present in the seed coats of a variety of plant species including vanilla orchid, various cacti, and the ornamental plant Cleome hassleriana . Because of its unique chemical and physical properties, there is considerable interest in engineering C-lignin into the cell walls of bioenergy crops as a high-value co-product of bioprocessing. We have used information from a transcriptomic analysis of developing C. hassleriana seed coats to suggest strategies for engineering C-lignin in a heterologous system, using hairy roots of the model legume Medicago truncatula . Results We systematically tested strategies for C-lignin engineering using a combination of gene overexpression and RNAi-mediated knockdown in the caffeic acid/5-hydroxy coniferaldehyde 3/5-O-methyltransferase ( comt ) mutant background, monitoring the outcomes by analysis of lignin composition and profiling of monolignol pathway metabolites. In all cases, C-lignin accumulation required strong down-regulation of caffeoyl CoA 3- O -methyltransferase (CCoAOMT) paired with loss of function of COMT. Overexpression of the Selaginella moellendorffii ferulate 5-hydroxylase ( SmF5H ) gene in comt mutant hairy roots resulted in lines that unexpectedly accumulated high levels of S-lignin. Conclusion C-Lignin accumulation of up to 15% of total lignin in lines with the greatest reduction in CCoAOMT expression required the strong down-regulation of both COMT and CCoAOMT, but did not require expression of a heterologous laccase, cinnamyl alcohol dehydrogenase (CAD) or cinnamoyl CoA reductase (CCR) with preference for 3,4-dihydroxy-substituted substrates in M. truncatula hairy roots. Cell wall fractionation studies suggested that the engineered C-units are not present in a heteropolymer with the bulk of the G-lignin.

09 BIOMASS FUELS↗

Population structure of Miscanthus sacchariflorus reveals two major polyploidization events, tetraploid-mediated unidirectional introgression from diploid M. sinensis, and diversity centred around the Yellow Sea

Abstract Background and Aims Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sacchariflorus has a broad geographic range, is used to produce paper in China and is one of the parents (along with Miscanthus sinensis) of the important biomass species Miscanthus × giganteus. The largest study of M. sacchariflorus population genetics to date is reported here. Methods Collections included 764 individuals across East Asia. Samples were genotyped with 34 605 single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RAD-seq) and ten plastid microsatellites, and were subjected to ploidy analysis by flow cytometry. Key Results Six major genetic groups within M. sacchariflorus were identified using SNP data: three diploid groups, comprising Yangtze (M. sacchariflorus ssp. lutarioriparius), N China and Korea/NE China/Russia; and three tetraploid groups, comprising N China/Korea/Russia, S Japan and N Japan. Miscanthus sacchariflorus ssp. lutarioriparius was derived from the N China group, with a substantial bottleneck. Japanese and mainland tetraploids originated from independent polyploidization events. Hybrids between diploid M. sacchariflorus and M. sinensis were identified in Korea, but without introgression into either parent species. In contrast, tetraploid M. sacchariflorus in southern Japan and Korea exhibited substantial hybridization and introgression with local diploid M. sinensis. Conclusions Genetic data indicated that the land now under the Yellow Sea was a centre of diversity for M. sacchariflorus during the last glacial maximum, followed by a series of migrations as the climate became warmer and wetter. Overall, M. sacchariflorus has greater genetic diversity than M. sinensis, suggesting that breeding and selection within M. sacchariflorus will be important for the development of improved M. × giganteus. Ornamental M. sacchariflorus genotypes in Europe and North America represent a very narrow portion of the species’ genetic diversity, and thus do not well represent the species as a whole.

Clark, Lindsay V.↗

Use of remote sensing in agriculture

The remote sensing studies of (a) cultivated peanut areas in Southeastern Virginia; (b) studies at the Virginia Truck and Ornamentals Research Station near Painter, Virginia, the Eastern Virginia Research Station near Warsaw, Virginia, the Tidewater Research and Continuing Education Center near Suffolk, Virginia, and the Southern Piedmont Research and Continuing Education Center Blackstone, Virginia; and (c) land use classification studies at Virginia Beach, Virginia are presented. The practical feasibility of using false color infrared imagery to detect and determine the areal extent of peanut disease infestation of Cylindrocladium black rot and Sclerotinia blight is demonstrated. These diseases pose a severe hazard to this major agricultural food commodity. The value of remote sensing technology in terrain analyses and land use classification of diverse land areas is also investigated. Continued refinement of spectral signatures of major agronomic crops and documentation of pertinent environmental variables have provided a data base for the generation of an agricultural-environmental prediction model.

Pettry, D. E.↗

Final Report DE-SC0006634. Quantifying phenotypic and genetic diversity of Miscanthus sinensis as a resource for knowledge-based improvement of M. ×giganteus (M. sinensis × M. sacchariflorus)

Miscanthus is especially attractive as a bioenergy crop for temperate environments because it produces high yields, needs few inputs, and grows well during the cool weather of early spring and late fall when few warm-season grasses can. However, Miscanthus feedstock production for the emerging U.S. bioenergy industry and for existing demand in Europe is based on a single sterile, vegetatively propagated variety of M. ×giganteus. M. ×giganteus is an interspecific hybrid of the parental species M. sinensis and M. sacchariflorus. Prior to the current study, little information existed about the genetic diversity and breeding potential of either M. ×giganteus parental species. In the current project, we studied more than 600 accessions of M. sinensis from throughout its native range in China, Japan, and Korea, in addition to ornamental cultivars and U.S. naturalized populations. Using thousands of DNA markers, we identified seven geographically distinct genetic groups of M. sinensis. Notably, we found that the ornamental cultivars and U.S. naturalized populations were derived from only a subset of the Southern Japan group, indicating that our study greatly increased the genetic diversity available for breeding new biomass cultivars. Additionally, this new understanding of M. sinensis population structure could be used to predict which crosses may produce progeny with the greatest hybrid vigor. Replicated field trials were also established at multiple locations in North America and Asia. Data on traits of importance for biomass productivity, such as flowering time, yield and height, were taken. Analyses of the phenotypic data from the field trials along with the DNA markers allowed us to identify many marker-trait associations. These results will enable marker-assisted breeding, which will allow selection at the seedling stage rather than waiting two to three years to obtain phenotypic data. Thus, this study is expected to greatly increase the efficiency of breeding Miscanthus for improved adaptation and biomass yield.

09 BIOMASS FUELS↗