Forest use suitability:Towards decision-making-oriented sustainable management of forest ecosystem services

Management of forest lands considering multi-functional approaches is the basis to sustain or enhance the provi-sion of specific benefits,while minimizing negative impacts to the environment.Defining a desired management itinerary to a forest depends on a variety of factors,including the forest type,its ecological characteristics,and the social and economic needs of local communities.A strategic assessment of the forest use suitability(FUS)(namely productive,protective,conservation-oriented,social and multi-functional)at regional level,based on the provision of forest ecosystem services and trade-offs between FUS alternatives,can be used to develop management strategies that are tailored to the specific needs and conditions of the forest.The present study assesses the provision of multiple forest ecosystem services and employs a decision model to identify the FUS that sup-ports the most present and productive ecosystem services in each stand in Catalonia.For this purpose,we apply the latest version of the Ecosystem Management Decision Support(EMDS)system,a spatially oriented decision support system that provides accurate results for multi-criteria management.We evaluate 32 metrics and 12 as-sociated ecosystem services indicators to represent the spatial reality of the region.According to the results,the dominant primary use suitability is social,followed by protective and productive.Nevertheless,final assignment of uses is not straightforward and requires an exhaustive analysis of trade-offs between all alternative options,in many cases identifying flexible outcomes,and increasing the representativeness of multi-functional use.The assignment of forest use suitability aims to significantly improve the definition of the most adequate management strategy to be applied.

Review Article: High-Temperature Adult-Plant Resistance, Key for Sustainable Control of Stripe Rust

High-temperature adult-plant (HTAP) resistance expresses when plants grow old and the weather becomes warm. This non-race specific and durable type of resistance has been used successfully in control of wheat stripe rust in the US since early 1960s. This article describes practical procedures for identification and characterization of HTAP resistance and reviews recent studies on discovery of genes conferring HTAP resistance. Recent studies providing insights to the molecular basis for the durability of HTAP resistance will be presented. Strategies for improving levels of HTAP resistance and improving control of stripe rust through combining HTAP resistance with effective all-stage resistance will be discussed.

Dual-Use Bioenergy-Livestock Feed Potential of Giant Miscanthus, Giant Reed, and Miscane

High yielding perennial grasses could integrate bioenergy-livestock operations, thereby, offsetting diversions of cropland to lignocellulosic crops, but research is needed to determine chemical composition and digestibility of leaf and stem fractions that might affect downstream reside uses. The objective of this study was to compare feedstock quality of leaf and stem tissues of dedicated bioenergy feedstocks: giant miscanthus (Miscanthus × giganteus), giant reed (Arundo donax), and miscane (Saccharum hybrid × Miscanthus spp.) when grown with or without supplemental irrigation on an upland site. Three species were space-planted on a silt loam soil in March 2007 and harvested prior to the first freeze in plant-cane, first ratoon, and second-ratoon crops for three years. Giant miscanthus leaf tissue had greatest acid detergent lignin and cellulose, and lowest concentrations of nitrogen (N) and total nonstructural carbohydrates (TNC) in ratoon crops. Giant reed leaf tissue had greatest concentrations of in vitro digestible dry matter (IVDMD), TNC, and N (P ≤ 0.05). Conversely, miscane stem tissue had greatest concentrations of IVDMD, TNC, hemicellulose, and low dry matter and combustible energy (P ≤ 0.05). Results suggest all species’ residue has positive feedstock attributes for thermochemical bioenergy conversion, and albeit giant miscanthus has very little potential value as fodder. Miscane stem and giant reed leaf tissue have potential value as livestock feed, although giant reed is not currently recommended for planting. Further research is needed on dietary composition, acceptability, voluntary intake, and live weight gain before any of these species are recommended as livestock feed sources.

A hybrid swarm population of Pinus densiflora × P. sylvestris inferred from sequence analysis of chloroplast DNA and morphological characters

To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural stands or experimental stations to study whether shoot apex morphology of 4-year old seedlings can be correlated with the sequence of a chloroplast DNA simple sequence repeat marker （cpDNA SSRs）. Total genomic DNA was extracted and subjected to sequence analysis of the pine cpDNA SSR marker Pt15169. Results show that morphological characters from 4-year old seedlings did not correlate with sequence variants of this marker. Marker haplotypes from all P. sylvestris trees had a CTAT element that was absent from all sampled P. densiflora trees. However, both haplotype classes involving this insertion/deletion element were found in a P. densiflora × P. sylvestris population and its seedling progeny. It was concluded that the P. densiflora × P. sylvestris accessions sampled from Jilin, China resulted from bi-directional crosses, as evidenced by both species’ cpDNA haplotypes within the hybrid swarm population.

Use of Cross-Fostering to Enhance Growth of Pigs That Are Predicted to Grow Poorly Based on Plasma α-1 Acid Glycoprotein Concentration

Porcine α-1 acid glycoprotein (AGP) in newborn pigs can be used to predict growth rate through weaning and is a marker for growth impairment. This study examined whether nutritional support can improve the growth rate of piglets identified as having poor growth potential. Cross-fostering (CF) and CF plus a milk supplement (CF + MS) were used to attempt to improve the growth performance of pigs. Blood was collected at d1 post-parturition for measurement of plasma AGP for all pigs in 28 litters contributing to the experiment. Piglets with the highest plasma AGP level were weight and sex matched to a littermate with a low plasma AGP concentration and four pairs of these weight and sex matched pigs were grouped into four foster litters per treatment (control, CF, CF + MS). The control group was assembled by pairing littermates remaining in donor litters. Pigs stayed on treatment until weaning at 21 days of age. At 35 days of age, dual energy X-ray absorptiometry (DXA) was performed on CF and CF + MS pigs to evaluate carcass composition. Control pairs differed in weaning weight, with pigs with higher plasma AGP at 1 day of age having smaller weaning weights than their littermates of similar birth weight (P < 0.05). However, CF eliminated the difference in weaning weight between the slow growing pigs and their birth weight matched littermates. CF + MS produced a similar effect as CF (P > 0.05). At 35 days of age, body weights were still similar between CF littermates and between CF + MS littermates (P > 0.05). DXA analysis demonstrated that body composition was similar between CF or CF + MS treated pigs and their littermates. These data demonstrate that CF can be used to correct the growth impairment in pigs predicted using plasma AGP as the marker. CF + MS can do the same, but at greater expense.

Growth Rates of Giant Miscanthus (<i>Miscanthus</i>×<i>giganteus</i>) and Giant Reed (<i>Arundo donax</i>) in a Low-Input System in Arkansas, USA

The US Department of Energy is currently building strategies for the expansion of clean and renewable energy sources, and tall, rapidly-growing grasses such as giant miscanthus (Miscanthus × giganteus) and giant reed (Arundo donax) are two of the many of species that could fill this renewable energy niche. The objective was to compare stalk growth components of giant miscanthus and giant reed, in a low-input system (no irrigation and no fertilizer use) in Arkansas, USA. Due to the potential invasiveness of giant reed, our study was conducted on an upland site to minimize escape. Plant height and dry weight per stalk were measured every week for two consecutive growing seasons in 2012 and 2013. Leaf area index (LAI) was measured every two weeks from May to September in 2012. A significant species × day interaction occurred for plant height and dry weight per stalk, due to the relatively greater height and weight of giant reed compared to giant miscanthus after May. Stalk elongation rate was greater for giant reed than giant miscanthus (1.85 and 1.11 cm day-1, respectively). Leaf area index differed between species, giant reed (10.4 m2 m-2) > giant miscanthus (4.4 m2 m-2). We showed that giant reed produced taller, heavier stalks, and had a greater stalk elongation rate, compared to giant miscanthus. For sustainable bioenergy production from giant reed in Arkansas, further studies should be performed to determine ideal number of harvests per year and associated production cost.

Astragalus polysaccharide enhances immunity and inhibits H9N2 avian influenza virus in vitro and in vivo

This study investigated the humoral immunization of Astragalus polysaccharide （APS） against HgN2 avian influenza virus （H9N2 AIV） infection in chickens. The effects of APS treatment on H9N2 infection was evaluated by an Mqq- [3（4, 5-dimethylthiazol-2-yl）-2, 3-diphenyl tetrazolium bromide] assay and analysis of MFIC and cytokine mRNA expression. The effect on lymphocyte and serum antibody titers in vivo was also investigated. IL-4, IL-6, IL-10, LITAF, IL-12 and antibody titers to H9N2 AIV wet enhanced in the first week after APS treatment. The results indicated that APS treatment reduces H9N2 AIV replication and promotes early humoral immune responses in young chickens.This study investigated the humoral immunization of Astragalus polysaccharide （APS） against HgN2 avian influenza virus （H9N2 AIV） infection in chickens. The effects of APS treatment on HgN2 infection was evaluated by an M]q- [3（4, 5-dimethylthiazol-2-yl）-2, 3-diphenyl tetrazolium bromide] assay and analysis of MHC and cytokine mRNA expression. The effect on lymphocyte and serum antibody titers in vivo was also investigated. IL-4, IL-6, IL-10, LITAF, IL-12 and antibody titers to PIgN2 AIV were enhanced in the first week after APS treatment. The results indicated that APS treatment reduces HgN2 AIV replication and promotes early humoral immune responses in young chickens.

Remote Sensing and Precision Agriculture Technologies for Crop Disease Detection and Management with a Practical Application Example

Remote sensing technology has long been used to detect and map crop diseases.Airborne and satellite imagery acquired during growing seasons can be used not only for early detection and within-season management of some crop diseases,but also for the control of recurring diseases in future seasons.With variable rate technology in precision agriculture,site-specific fungicide application can be made to infested areas if the disease is stable,although traditional uniform application is more appropriate for diseases that can spread rapidly across the field.This article provides a brief overview of remote sensing and precision agriculture technologies that have been used for crop disease detection and management.Specifically,the article illustrates how airborne and satellite imagery and variable rate technology have been used for detecting and mapping cotton root rot,a destructive soilborne fungal disease,in cotton fields and how site-specific fungicide application has been implemented using prescription maps derived from the imagery for effective control of the disease.The overview and methodologies presented in this article should provide researchers,extension personnel,growers,crop consultants,and farm equipment and chemical dealers with practical guidelines for remote sensing detection and effective management of some crop diseases.

Gene expression profiling of MYC-driven tumor signatures in porcine liver stem cells by transcriptome sequencing

AIM:To identify the genes induced and regulated by the MYC protein in generating tumors from liver stem cells.METHODS:In this study,we have used an immortal porcine liver stem cell line,PICM-19,to study the role of c-MYC in hepatocarcinogenesis.PICM-19 cells were converted into cancer cells(PICM-19-CSCs)by overexpressing human MYC.To identify MYC-driven differential gene expression,transcriptome sequencing was carried out by RNA sequencing,and genes identified by this method were validated using real-time PCR.In vivo tumorigenicity studies were then conducted by injecting PICM-19-CSCs into the flanks of immunodeficient mice.RESULTS:Our results showed that MYC-overexpressing PICM-19 stem cells formed tumors in immunodeficient mice demonstrating that a single oncogene was sufficient to convert them into cancer cells(PICM-19-CSCs).By using comparative bioinformatics analyses,we have determined that>1000 genes were differentially expressed between PICM-19 and PICM-19-CSCs.Gene ontology analysis further showed that the MYCinduced,altered gene expression was primarily associated with various cellular processes,such as metabolism,cell adhesion,growth and proliferation,cell cycle,inflammation and tumorigenesis.Interestingly,six genes expressed by PICM-19 cells(CDO1,C22orf39,DKK2,ENPEP,GPX6,SRPX2)were completely silenced after MYC-induction in PICM-19-CSCs,suggesting that the absence of these genes may be critical for inducingtumorigenesis.CONCLUSION:MYC-driven genes may serve as promising candidates for the development of hepatocellular carcinoma therapeutics that would not have deleterious effects on other cell types in the liver.

Eggshell and environmental bacteria contribute to the intestinal microbiota of growing chickens

Background: The initial intestinal microbiota acquired from different sources has profound impacts on animal health and productivity. In modern poultry production practices, the source(s) of the establishing microbes and their overall contribution during development of gastrointestinal tract communities are still unclear. Using fertilized eggs from two independent sources, we assessed the impact of eggshell-and environmental-associated microbial communities on the successional processes and bacterial community structure throughout the intestinal tract of chickens for up to 6 weeks post-hatch.Results: Culturing and sequencing techniques identified a viable, highly diverse population of anaerobic bacteria on the eggshell. The jejunal, ileal, and cecal microbial communities for the egg-only, environment-only, and conventionally raised birds generally displayed similar successional patterns characterized by increasing community richness and evenness over time, with strains of Enterococcus, Romboutsia, and unclassified Lachnospiraceae abundant for all three input groups in both trials. Bacterial community structures differed significantly based on trial and microbiota input with the exception of the egg-exposed and conventional birds in the jejunum at week 1 and the ileum at week 6. Cecal community structures were different based on trial and microbiota input source, and cecal short-chain fatty acid profiles at week 6 highlighted functional differences as well.Conclusion: We identified distinct intestinal microbial communities and differing cecal short-chain fatty acid profiles between birds exposed to the microbiota associated with either the eggshell or environment, and those of conventionally hatched birds. Our data suggest the eggshell plays an appreciable role in the development of the chicken intestinal microbiota, especially in the jejunum and ileum where the community structure of the eggshellonly birds was similar to the structure of conventionally hatched birds. Our data identify a complex interplay between the eggshell and environmental microbiota during establishment and succession within the chicken gut.Further studies should explore the ability of eggshell-and environment-derived microbes to shape the dynamics of succession and how these communities can be targeted through interventions to promote gut health and mitigate food-borne pathogen colonization in poultry.

Understanding development and ripening of fruit crops in an‘omics’ era

Next generation sequencing has revolutionized plant biology.Not only has our understanding of plant metabolism advanced using model systems and modern chromatography,but application of‘omics’-based technology has been widely extended to non-model systems as costs have plummeted and efficiency increased.As a result,important fundamental questions relating to important horticultural crops are being answered,and novel approaches with application to industry are in progress.Here we review recent research advances on development and ripening of fruit crops,how next generation sequencing approaches are driving this advance and the emerging future landscape.

Neosporosis: An Overview of Its Molecular Epidemiology and Pathogenesis

Neospora caninum (N. caninum), a cyst-forming protozoan parasite, is a major cause of bovine abortions and neonatal mortality worldwide. N. caninum has a broad intermediate host range, and its sexual cycle occurs exclusively in canids. Another species of Neospora, Neospora hughesi (N. hughesi), has been identified and causes myeloencephalitis in horses. Although molecular epidemiology studies are in their infancy, the 18S ribosomal RNA (rRNA) and ITS1 regions within the small subunit ribosomal RNA (ssuRNA) and an N. caninum species-specific DNA probe (pNc5) have been used extensively to differentiate Neospora from other closely related apicomplexan parasites. While these repetitive regions have higher sensitivity and specificity than housekeeping or antigen genes, they suffer from low discriminatory power and fail to capture intra-species diversity. Similarly, although multiple minisatellite or microsatellite marker studies have shown clear geographic substructures within Neospora, strains are often misclassified due to a convergence in the size of different alleles at microsatellite loci, known as homoplasy. Only one strain, N. caninum Liverpool (Nc-Liv), has been genome sequenced and compared with its closest relative, Toxoplasma gondii (T. gondii). Hence, detailed population genomics studies based on whole-genome sequences from multiple strains worldwide are needed in order to better understand the current population genetic structure of Neospora, and ultimately to determine more effective vaccine candidates against bovine neosporosis. The aim of this review is to outline our current understanding of the molecular epidemiology and genomics of Neospora in juxtaposition with the closely related apicomplexan parasites Hammondia hammondi and T. gondii.

Review of oxidative stress and antioxidative defense mechanisms in Gossypium hirsutum L.in response to extreme abiotic conditions

Oxidative stress occurs when crop plants are exposed to extreme abiotic conditions that lead to the excessive production and accumulation of reactive oxygen species(ROS).Those extreme abiotic conditions or stresses include drought,high temperature,heavy metals,salinity,and ultraviolet radiation,and they cause yield and quality losses in crops.ROS are highly reactive species found in nature that can attack plant organelles,metabolites,and molecules by interrupting various metabolic pathways until cell death occurs.Plants have evolved defense mechanisms for the production of antioxidants to detoxify the ROS and to protect the plant against oxidative damage.Modern researches in crop plants revealed that low levels of ROS act as a signal which induces tolerance to environmental extremes by altering the expression of defensive genes.In this review,we summarized the processes involved in ROS production in response to several types of abiotic stress in cotton plants.Furthermore,we discussed the achievements in the understanding and improving oxidative stress tolerance in cotton in recent years.Researches related to plant oxidative stresses have shown excellent potential for the development of stress-tolerant crops.

Identification of a Highly Expressed 3-Hydroxy-3-Methylglutaryl-CoA Reductase Gene in the Root Tissue of <i>Taraxacum kok-saghyz</i>

Kazakh dandelion (Taraxacum kok-saghyz, Tk) is a rubber-producing plant currently being investigated as a source of natural rubber for industrial applications. Like many other isoprenoids, rubber is a downstream product of the mevalonate pathway. The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) enzyme catalyzes the conversion of 3-hydroxy-3-methylglutaryl-CoA to mevalonic acid, a key regulatory step in the MVA pathway. Such regulated steps provide targets for increases in isoprenoid and rubber contents via genetic engineering to increase enzyme activities. In this study, we identify a TkHMGR1 gene that is highly expressed in the roots of Kazakh dandelion, the main tissue where rubber is synthesized and stored. This finding paves the way for further molecular and genetic studies of the TkHMGR1 gene, and its role in rubber biosynthesis in Tk and other rubber-producing plants.

Association Analysis and Identification of SNP Markers for Stemphylium Leaf Spot (Stemphylium botryosum f. sp. spinacia) Resistance in Spinach (Spinacia oleracea)

Stemphylium leaf spot, caused by Stemphylium botryosum f. sp. spinacia, is an important fungal disease of spinach (Spinacia oleracea L.). The aim of this study was to conduct association analysis to identify single nucleotide polymorphism (SNP) markers associated with Stemphylium leaf spot resistance in spinach. A total of 273 spinach genotypes, including 265 accessions from the USDA spinach germplasm collection and eight commercial cultivars, were used in this study. Phenotyping for Stemphylium leaf spot resistance was evaluated in greenhouse;genotyping was conducted using genotyping by sequencing (GBS) with 787 SNPs;and single marker regression, general linear model, and mixed linear model were used for association analysis of Stemphylium leaf spot. Spinach genotypes showed a skewed distribution for Stemphylium leaf spot resistance, with a range from 0.2% to 23.5% disease severity, suggesting that Stemphylium leaf spot resistance in spinach is a complex, quantitative trait. Association analysis indicated that eight SNP markers, AYZV02052595_115, AYZV02052595_122, AYZV02057770_10404, AYZV02129827_205, AYZV0-2152692_182, AYZV02180153_337, AYZV02225889_197, and AYZV02258563_213 were strongly associated with Stemphylium leaf spot resistance, with a Log of the Odds (LOD) of 2.5 or above. The SNP markers may provide a tool to select for Stemphylium leaf spot resistance in spinach breeding programs through marker-assisted selection (MAS).

A community resource for exploring and utilizing genetic diversity in the USDA pea single plant plus collection

Globally,pea(Pisum sativum L.)is an important temperate legume crop for food,feed and fodder,and many breeding programs develop cultivars adapted to these end-uses.In order to assist pea development efforts,we assembled the USDA Pea Single Plant Plus Collection(PSPPC),which contains 431 P.sativum accessions with morphological,geographic and taxonomic diversity.The collection was characterized genetically in order to maximize its value for trait mapping and genomics-assisted breeding.To that end,we used genotyping-by-sequencing—a cost-effective method for de novo single-nucleotide polymorphism(SNP)marker discovery—to generate 66591 high-quality SNPs.These data facilitated the identification of accessions divergent from mainstream breeding germplasm that could serve as sources of novel,favorable alleles.In particular,a group of accessions from Central Asia appear nearly as diverse as a sister species,P.fulvum,and subspecies,P.sativum subsp.elatius.PSPPC genotypes can be paired with new and existing phenotype data for trait mapping;as proof-of-concept,we localized Mendel’s A gene controlling flower color to its known position.We also used SNP data to define a smaller core collection of 108 accessions with similar levels of genetic diversity as the entire PSPPC,resulting in a smaller germplasm set for research screening and evaluation under limited resources.Taken together,the results presented in this study along with the release of a publicly available SNP data set comprise a valuable resource for supporting worldwide pea genetic improvement efforts.

Screening of Seed Soluble Sugar Content in Cowpea (<i>Vigna unguiculata</i>(L.) Walp)

Cowpea (Vigna unguiculata (L.) Walp) is a legume crop grown worldwide to provide protein, starch, soluble sugar, amino acids, fatty acids, minerals, and vitamins for human consumption and animal feed. Soluble sugar is an important physiological trait in cowpea seeds. It not only plays an important role in storability and abiotic stress tolerance in seeds, but also provides energy to the human body and improves cooking quality by giving a desirable taste and mouth feel. Therefore, this research aimed to evaluate variation of cowpea seed soluble sugar content, and assess the soluble sugar content of cowpea varieties with different seed coat colors for cowpea consumers. A total of 113 cowpea genotypes were used, including 89 USDA GRIN germplasm accessions and 24 advanced breeding lines developed from University of Arkansas, AR, USA. Seed soluble sugar content was measured using a Spectrophotometer by phenol-sulphuric acid method. The results showed that the seed soluble sugar contents among 113 cowpea genotypes exhibited a wide range varying from 32.6 mg/g to 86.1 mg/g with an average of 54.5 mg/g. The five highest soluble sugar contents in seeds were found in the Arkansas cultivar Empire (86.1 mg/g), USDA germplasm accession PI583202 (84.5 mg/g), Arkansas advanced breeding line 09-655 (82.1 mg/g), USDA accession PI601085 (81.6 mg/g), and Arkansas advanced breeding line 09-529 (80.9 mg/g), and they can be used in cowpea breeding programs to develop new cowpea cultivars with higher seed soluble sugar content. It was also observed that the soluble sugar contents in Arkansas commercial cultivars and advanced breeding lines were higher than those in USDA germplasm and the seeds with colorful coat may have soluble sugar levels desirable by consumers.

Role of SNPs in determining QTLs for major traits in cotton

A single nucleotide polymorphism is the simplest form of genetic variation among individuals and can induce minor changes in phenotypic,physiological and biochemical characteristics.This polymorphism induces various mutations that alter the sequence of a gene which can lead to observed changes in amino acids.Several assays have been developed for identification and validation of these markers.Each method has its own advantages and disadvantages but genotyping by sequencing is the most common and most widely used assay.These markers are also associated with several desirable traits like yield,fibre quality,boll size and genes respond to biotic and abiotic stresses in cotton.Changes in yield related traits are of interest to plant breeders.Numerous quantitative trait loci with novel functions have been identified in cotton by using these markers.This information can be used for crop improvement through molecular breeding approaches.In this review,we discuss the identification of these markers and their effects on gene function of economically important traits in cotton.

Pigmentation Restored in Mutant Laboratory Strain of the Lady Beetle Coleomegilla maculata through Dietary Supplementation

A laboratory colony of Coleomegilla maculata (DeGeer), ye, selected for a pigmentation deficiency, was restored to near wild type cuticle coloration by adding crushed heads and wings of the red colored parental strain to the diet. While the wings and other colored portions of the cuticle re-gained the red color, the eyes of the pigmentation deficient insects were not changed from the pale mutant form. Plant derived carotenes lycopene and beta-carotene did not restore the mutant beetles to a visibly distinguishable red color. An additional pigmentation deficient mutant strain, gold, partially recovered red cuticle color when provided with diet containing pigmented insect particles. This work represents the first rescue of a color phenotype in a lady beetle.

White (<i>Trifolium repens</i>L.) and Arrowleaf (<i>Trifolium vesiculosum</i>Savi) Clover Emergence in Varying Loblolly Pine (<i>Pinus taeda</i>L.) Tree Alley Spacings

Agroforestry systems have the potential to provide year-long income opportunities via the integrated forage or crop, timber, and livestock. Legumes are an attractive alternative option during the growing season when more traditional forages may not be as productive. The objective of this study was to test the establishment of arrowleaf and white clover grown under varying pine tree alley widths. In 2011, existing forage was removed in 15-yr old loblolly pine tree row alleys of different widths (3.7, 4.9, 7.3, and 9.8 m), including an open area. Arrowleaf, as an annual, was replanted in 2012. Seedlings were counted twice/year, while dry matter was measured three times/year. Photosynthetically active radiation (PAR) was measured in all alley widths to compare light penetration through the canopy. Hot and dry conditions occurred throughout 2012, affecting results. In 2012 and 2013, the greatest PAR for most treatments was observed in June. Seedling counts for all treatments were greatest immediately after establishment, and gradually declined throughout the course of the study. Dry matter yields increased throughout the growing season, and were greatest in arrowleaf clover in the open area on all measurement dates;however, increased weed pressure and repeated flooding affected yields. This study demonstrated that clover establishment in shady wooded areas is possible, but only under suitable environmental conditions.