Tailoring poplar lignin without yield penalty by combining a null and haploinsufficient CINNAMOYL-CoA REDUCTASE2 allele

Lignin causes lignocellulosic biomass recalcitrance to enzymatic hydrolysis. Engineered low-lignin plants have reduced recalcitrance but often exhibit yield penalties, offsetting their gains in fermentable sugar yield. Here, CRISPR/Cas9-generated CCR2(−/*) line 12 poplars have one knockout CCR2 allele while the other contains a 3-bp deletion, resulting in a 114I115A-to-114T conversion in the corresponding protein. Despite having 10% less lignin, CCR2(−/*) line 12 grows normally. On a plant basis, the saccharification efficiency of CCR2(−/*) line 12 is increased by 25–41%, depending on the pretreatment. Analysis of monoallelic CCR2 knockout lines shows that the reduced lignin amount in CCR2(−/*) line 12 is due to the combination of a null and the specific haploinsufficient CCR2 allele. Analysis of another CCR2(−/*) line shows that depending on the specific CCR2 amino-acid change, lignin amount and growth can be affected to different extents. Our findings open up new possibilities for stably fine-tuning residual gene function in planta.

A Native Bee, Melissodes tepaneca (Hymenoptera: Apidae), Benefits Cotton Production

The cotton agroecosystem is one of the most intensely managed, economically and culturally important cropping systems worldwide. Native pollinators are essential in providing pollination services to a diverse array of crops, including those which have the ability to self-pollinate. Cotton, which is autogamous, can potentially benefit from insect-mediated pollination services provided by native bees within the agroecosystem. Examined through two replicated experiments over two years, we hypothesized that native bees facilitated cross-pollination, which resulted in increased lint of harvested bolls produced by flowers exposed to bees and overall lint weight yield of the plant. Cotton bolls from flowers that were caged and exposed to bees, flowers that were hand-crossed, and bolls from flowers on uncaged plants exposed to pollinators had higher pre-gin weights and post-gin weights than bolls from flowers of caged plants excluded from pollinators. When cotton plants were caged with the local native bee Melissodes tepaneca, seed cotton weight was 0.8 g higher on average in 2018 and 1.18 g higher on average in 2019 than when cotton plants were excluded from bees. Cotton production gains from flowers exposed to M. tepaneca were similar when measuring lint and seed separately. Cotton flowers exposed over two weeks around the middle of the blooming period resulted in an overall yield gain of 12% to 15% on a whole plant basis and up to 24% from bolls produced from flowers exposed directly to M. tepaneca. This information complements cotton-mediated conservation benefits provided to native pollinators by substantiating native bee-mediated pollination services provided to the cotton agroecosystem.

Distribution of Lanthanides, Yttrium, and Scandium in the Pilot-Scale Beneficiation of Fly Ashes Derived from Eastern Kentucky Coals

In this study, Central Appalachian coal-derived fly ashes from two power plants were beneficiated in a pilot-scale facility in order to produce a product with a relatively consistent concentration of rare earth elements (REE). The <200-mesh final fly ash product was produced by removing the carbon- and Fe-rich particles prior to screening at 200 mesh (75 µm). The Plant D fly ash had high concentrations of CaO and SO3, which were diminished through the two months when the ash was being beneficiated, representing a consequence of the heat, humidity, and excessive rainfall in the Kentucky summer. The high CaO and SO3 concentrations through the early runs likely contributed to the lower REE in the <200-mesh products of those runs. Of the non-REE minor elements, Ba, V, Mn, Zn, and As showed the greatest between-run variations within the runs for each plant. The overall REE concentrations proved to be similar, both on a between-run basis for the individual fly ash sources and on a between-plant basis. Variations in fly ash quality will occur in larger-scale operations, so on-going attention to the fly ash quality and the response of the fly ash to beneficiation is necessary. Changes in the Plant D fly ash with time imply that both the freshness of the original ash and the length and conditions of its storage at the site of beneficiation could be factors in the quality and consistency of the processed fly ash.

Regulation of Symbiotic Nitrogen Fixation in Legume Root Nodules

In most legume nodules, the di-nitrogen (N2)-fixing rhizobia are present as organelle-like structures inside their root host cells. Many processes operate and interact within the symbiotic relationship between plants and nodules, including nitrogen (N)/carbon (C) metabolisms, oxygen flow through nodules, oxidative stress, and phosphorous (P) levels. These processes, which influence the regulation of N2 fixation and are finely tuned on a whole-plant basis, are extensively reviewed in this paper. The carbonic anhydrase (CA)-phosphoenolpyruvate carboxylase (PEPC)-malate dehydrogenase (MDH) is a key pathway inside nodules involved in this regulation, and malate seems to play a crucial role in many aspects of symbiotic N2 fixation control. How legumes specifically sense N-status and how this stimulates all of the regulatory factors are key issues for understanding N2 fixation regulation on a whole-plant basis. This must be thoroughly studied in the future since there is no unifying theory that explains all of the aspects involved in regulating N2 fixation rates to date. Finally, high-throughput functional genomics and molecular tools (i.e., miRNAs) are currently very valuable for the identification of many regulatory elements that are good candidates for accurately dissecting the particular N2 fixation control mechanisms associated with physiological responses to abiotic stresses. In combination with existing information, utilizing these abundant genetic molecular tools will enable us to identify the specific mechanisms underlying the regulation of N2 fixation.

Clinical-diagnostic criteria and peculiarities of treatment of urocystitis in cats

The results of a study of cats with signs of urocystitis are presented. The general principles of diagnostics based on clinical, laboratory and additional research methods are established. Clinically acute urocystitis was manifested by disorders of urination – pollakiuria, dysuria, stranguria. Some of the animals showed an increase in body temperature, pain in the bladder and tightness of the walls of the abdominal cavity. According to the results of urine study, it was observed that 43.7% of the cats with urocystitis showed an increase in relative density of the urine, a change in urine pH towards higher alkaline levels, urine residue – erythrocytes, leukocytes and microorganisms were indicated. Impurities of salt and sand were found in 58.4% of the cats. In 31.4% of the cats, urocystitis was a complication of urolithiasis. Early diagnosis, especially differential diagnosis of acute and chronic, secondary urocystitis presents difficulties. Therefore, in cases of suspected inflammatory diseases of the urinary tract, a complex examination using ultrasound of the kidneys, bladder, prostate gland (in males) and the uterus (in females) is recommended.With therapeutic purpose, sick cats (two groups of 8 animals/group) were administered: anti-microbial agent (5% solution of Enrofloxacin), spasmolytic medicinal agent – Riabal®. Animals of the second group were additionally administered the drug Cystocure (Candioli Farmaceutici). According to the instructions, the plant basis of the powder Cystocure is presented by extracts of cranberries and orange peels. These substances provide the main properties of the drug – help in resisting the bacterial colonization of the lower urinary tract and change in urine pH to the acid side. The applied therapy positively influenced the clinical condition of the cats. Their general health was improved, painful sensations during the act of urination disappeared, the quantity and frequency of diuresis normalized. On the seventh day of treatment, according to the results of the study, the urine became transparent, no microorganisms were detected. In cats of the second group, which additionally used the drug Cystocure, the amount of salts and sand decreased significantly, the pH of the urine did not exceed the recommended value of 6.4, in contrast to animals of the first group.

Impact of Growing Environment on Anthracnose Severity of Switchgrass Cultivars and Clones

Anthracnose (caused by Colletotrichum navitas) has the potential to significantly reduce biomass yield of switchgrass (Panicum virgatum L.); however, limited information is available on the impact of growing environment on tolerance of switchgrass to anthracnose. Therefore, the major objectives of this study were to (i) examine genotype–environment (G × E) effects on anthracnose severity in populations of switchgrass cultivars and individual genotypes and (ii) determine clonal repeatability estimates and stability analysis of anthracnose tolerance on individual switchgrass genotypes. Two experiments were conducted at one prime and two marginal soil locations in New Jersey. In all, 14 switchgrass cultivars were established from seed in 2008 for experiment 1 and 50 replicated switchgrass clones were planted in 2009 for experiment 2 at all three locations. Anthracnose was rated visually in 2010 for experiment 1 and in 2010 and 2011 for experiment 2. Significant G × E interactions were detected for both experiments (P ≤ 0.05) and anthracnose severity varied by location and cultivar. Clonal repeatability estimates for disease tolerance among clones was 0.78 on a clonal basis and 0.32 on a single-plant basis. Lowland ecotypes exhibited less disease overall than upland ecotypes. Results from this study indicate that selection for improved tolerance to anthracnose should be conducted after evaluation across several environments over multiple years.

Validation of activated sludge plant performance for virus and protozoan reduction

Validation studies were undertaken at Adelaide metropolitan wastewater treatment plants to establish the actual log10 reduction values (LRVs) of pathogens (viruses and Cryptosporidium) across activated sludge plants (ASPs) as an alternative to accepting the default values attributed by the Department of Health and Ageing (DHA). Grab samples were collected across a 6-week period and assessed for pathogens (adenovirus and Cryptosporidium) and indicator microorganisms (sulphite-reducing clostridia and F-RNA bacteriophage). Through applying the validation process, the DHA has revised the default value for reduction of viruses with an increase from 0.5 log10 to 1 log10 while the value for protozoa remains at 0.5 log10 based on the combined data for a well-operated and maintained ASP. This provides the basis for considering further work at individual plants which may allow higher log credits to be obtained on a plant by plant basis.

Effects of Soybean Leaf and Plant Age on Susceptibility to Initiation of Infection by Phakopsora pachyrhizi

To help resolve the question of relationship of soybean leaf and plant age to susceptibility to infection by Phakopsora pachyrhizi, we inoculated 50-day-old plants of cv. Williams 82 with a suspension at 3 × 103 urediniospores/ml, placed in growth chambers under a range of temperature conditions, and later examined for number of lesions. In addition, plants ranging in age from 20 days to 79 days were inoculated, placed in a greenhouse, and examined by leaf for number of lesions. Results showed that Williams 82 did not vary in susceptibility among leaves at different positions regardless of temperature, or among leaves at different positions (up to 11 trifoliolates) on individual plants ranging in age from 20 to 79 days at inoculation. However, on a per-plant basis, there was a gradual decrease in numbers of lesions as plants increased in age at inoculation. For example, whereas 37 lesions/cm2 leaf area were produced on 20-day-old plants, only 10 were produced on 79-day-old plants. Although the study does not rule out the possibility that there might be cultivar dependent differences in age response, it does strengthen the possibility that older plants are less susceptible, and that the increase in disease observed in the field during fall months may be due to an increase in favorability of the environment for disease or increase in inoculum pressure. Accepted for publication 23 November 2011. Published 27 February 2012.

Nitrogen Uptake and Mobilization by Hydrangea Leaves from Foliar-sprayed Urea in Fall Depend on Plant Nitrogen Status

Rooted liners of Hydrangea macrophylla (Thunb.) Ser. ‘Berlin’ were fertigated with different rates of nitrogen (N) from July to Sept. 2007 and leaves were sprayed with 15N-labeled urea in late October to evaluate urea uptake and 15N translocation by hydrangea leaves in relation to plant N status. Four plants from each N fertigation rate were harvested before they were sprayed with urea and 2, 5, 10, and 15 days after urea spray. Increasing rate of N fertigation increased plant N content in October before being sprayed with urea. Leaves rapidly absorbed 15N from urea spray. The highest rate of 15N uptake occurred during the first 2 days after urea spray and then decreased. Export of 15N from leaves occurred rapidly after uptake and the highest rate of 15N export occurred during the first 2 days after urea spray and then decreased. During the first 5 days after urea spray, the rate of 15N uptake by leaves and export from leaves decreased with increasing rate of N fertigation. On a whole plant basis, the total amount of 15N from foliar 15N–urea spray increased with increasing rate of N fertigation; however, the percentage of 15N exported from leaves and the percentage of N that derived from foliar 15N–urea spray decreased with increasing rate of N fertigation. Results suggest that hydrangea plants with lower N status in the fall are more efficient in absorbing and translocating N from foliar urea than plants with higher N status.