Role of CAMK2D in neurodevelopment and associated conditions.

The calcium/calmodulin-dependent protein kinase type 2 (CAMK2) family consists of four different isozymes, encoded by four different genes-CAMK2A, CAMK2B, CAMK2G, and CAMK2D-of which the first three have been associated recently with neurodevelopmental disorders. CAMK2D is one of the major CAMK2 proteins expressed in the heart and has been associated with cardiac anomalies. Although this CAMK2 isoform is also known to be one of the major CAMK2 subtypes expressed during early brain development, it has never been linked with neurodevelopmental disorders until now. Here we show that CAMK2D plays an important role in neurodevelopment not only in mice but also in humans. We identified eight individuals harboring heterozygous variants in CAMK2D who display symptoms of intellectual disability, delayed speech, behavioral problems, and dilated cardiomyopathy. The majority of the variants tested lead to a gain of function (GoF), which appears to cause both neurological problems and dilated cardiomyopathy. In contrast, loss-of-function (LoF) variants appear to induce only neurological symptoms. Together, we describe a cohort of individuals with neurodevelopmental disorders and cardiac anomalies, harboring pathogenic variants in CAMK2D, confirming an important role for the CAMK2D isozyme in both heart and brain function.

Short- and long-term effects of season-long infestation of Lycorma delicatula (Hemiptera: Fulgoridae) on young apple (Malus domestica) and peach (Prunus persica) trees.

Lycorma delicatula White, commonly called spotted lanternfly, is an invasive fulgorid that was first detected in the United States in Pennsylvania in 2014 and has spread across Eastern states. Lycorma delicatula is a phloem feeder, and feeding of persistent populations can cause wilt and dieback in host plants. Vineyards in infested regions have experienced vine loss, where damage has been attributed to L. delicatula. Nymphal and adult lifestages have been observed on cultivated apple and peach trees, but the feeding effects of this pest on these crops have not been quantified. Here, we caged young potted apple and peach trees and infested them with 50 nymphal or 25 adult L. delicatula per plant for 3 months and tracked plant health. Horticultural measurements including trunk diameter, height, leaf drop, and photosynthesis rates were recorded before, during, and after the infestation. To evaluate the long-term effects of infestation, all apple and peach trees were removed from pots and planted in an orchard block where their phenology, growth, and physiology could be evaluated the following growing season. Short- and long-term measurements showed no significant differences in apple tree health between infested and noninfested trees. There was a significant reduction in the growth of trunk diameter in peach trees during exposure to early-instar nymphs; infested peach trees also had significantly increased frost damage to buds the following spring. However, there were no long-term impacts on peach growth and physiology. These results indicate that L. delicatula likely will not be a major threat to these deciduous fruit crops.

Assessing acceptability of wild and cultivated hosts of Lycorma delicatula (Hemiptera: Fulgoridae) under semifield conditions.

Lycorma delicatula White (Hemiptera: Fulgoridae) is an invasive phloem feeder with a broad host range that includes both hardwood trees and cultivated temperate fruit crops. Here, we evaluated acceptability of wild hosts, A. altissima and Juglans nigra L. (Fagales: Juglandaceae) and cultivated hosts Vitis vinifera, Malus domestica (Rosales: Rosaceae), and Prunus persica L. Batsch (Rosales: Rosaceae) to L. delicatula under field conditions. Fluorescent-marked early instar nymphs, late instar nymphs, or adult L. delicatula were released at the base of single potted host plants and the number of individuals retained was recorded over 24 h. Paired choice trials with A. altissima and another host plant were conducted. Individuals retained on or moving between plants were recorded over 24 h. Sentinel A. altissima, J. nigra, V. vinifera, M. domestica, and P. persica potted plants were deployed at 5 sites and the number of L. delicatula present on each plant was recorded weekly. In single and paired host trials, early instars and adults were generally retained in higher numbers on A. altissima and V. vinifera, and late instars were retained on A. altissima and J. nigra. Significantly more L. delicatula were present on sentinel A. altissima compared with other host plants, except J. nigra during the period when late instars were the most prevalent lifestage in the field. These results indicate that wild hosts such as A. altissima and J. nigra are likely supporting establishment of L. delicatula populations, and that presence of cultivated V. vinifera may contribute to population establishment and growth.

Engineered gamma radiation phytosensors for environmental monitoring.

Nuclear energy, already a practical solution for supplying energy on a scale similar to fossil fuels, will likely increase its footprint over the next several decades to meet current climate goals. Gamma radiation is produced during fission in existing nuclear reactors and thus the need to detect leakage from nuclear plants, and effects of such leakage on ecosystems will likely also increase. At present, gamma radiation is detected using mechanical sensors that have several drawbacks, including: (i) limited availability; (ii) reliance on power supply; and (iii) requirement of human presence in dangerous areas. To overcome these limitations, we have developed a plant biosensor (phytosensor) to detect low-dose ionizing radiation. The system utilizes synthetic biology to engineer a dosimetric switch into potato utilizing the plant's native DNA damage response (DDR) machinery to produce a fluorescent output. In this work, the radiation phytosensor was shown to respond to a wide range of gamma radiation exposure (10-80 Grey) producing a reporter signal that was detectable at >3 m. Further, a pressure test of the top radiation phytosensor in a complex mesocosm demonstrated full function of the system in a 'real world' scenario.

Evaluating deployment strategies for spotted lanternfly (Lycorma delicatula Hemiptera: Fulgoridae) traps.

The spotted lanternfly, Lycorma delicatula (White) (Hemiptera: Fulgoridae), is an invasive planthopper that was first detected in the United States in Berks County, PA, in 2014, and has since spread to 13 states in the Eastern United States. This phloem-feeding pest has a broad host range, including economically important crops such as grapevine, Vitis spp. Monitoring presence and relative abundance of L. delicatula is essential to develop pest management tools. Here, we compared deployment strategies to optimize use of L. delicatula monitoring traps. Standard circle traps, sticky bands, and circle traps with replaceable bag tops were deployed at sites with either high or low populations present. Trap deployment at different heights and on different host tree species and trap sampling intervals were evaluated for standard circle traps only. Circle traps captured significantly more L. delicatula adults at low-density sites compared with other trap types in 2021, and no differences were detected at high-density sights. Traps deployed 1 m from the ground captured significantly more adults than those deployed at 0.5 m; no differences were detected for nymphs. While no significant differences in captures were found among intervals, weekly or biweekly sampling prevented specimen degradation. Although traps deployed on Ailanthus altissima (Mill.) Swingle (Sapindales: Simaroubaceae) captured significantly or numerically more L. delicatula at most sites, traps deployed on other hosts also yielded consistent captures. We were also able to alter the construction of circle trap skirts to allow for deployment on different sized tree trunks.

Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato.

Phytosensors are genetically engineered plant-based sensors that feature synthetic promoters fused to reporter genes to sense and report the presence of specific biotic and abiotic stressors on plants. However, when induced reporter gene output is below detectable limits, owing to relatively weak promoters, the phytosensor may not function as intended. Here, we show modifications to the system to amplify reporter gene signal by using a synthetic transcription factor gene driven by a plant pathogen-inducible synthetic promoter. The output signal was unambiguous green fluorescence when plants were infected by pathogenic bacteria. We produced and characterized a phytosensor with improved sensing to specific bacterial pathogens with targeted detection using spectral wavelengths specific to a fluorescence reporter at 3 m standoff detection. Previous attempts to create phytosensors revealed limitations in using innate plant promoters with low-inducible activity since they are not sufficient to produce a strong detectable fluorescence signal for standoff detection. To address this, we designed a pathogen-specific phytosensor using a synthetic promoter-transcription factor system: the S-Box cis-regulatory element which has low-inducible activity as a synthetic 4xS-Box promoter, and the Q-system transcription factor as an amplifier of reporter gene expression. This promoter-transcription factor system resulted in 6-fold amplification of the fluorescence after infection with a potato pathogen, which was detectable as early as 24 h post-bacterial infection. This novel bacterial pathogen-specific phytosensor potato plant demonstrates that the Q-system may be leveraged as a powerful orthogonal tool to amplify a relatively weak synthetic inducible promoter, enabling standoff detection of a previously undetectable fluorescence signal. Pathogen-specific phytosensors would be an important asset for real-time early detection of plant pathogens prior to the display of disease symptoms on crop plants.

Variation in cloud immersion, not precipitation, drives leaf trait plasticity and water relations in vascular epiphytes during an extreme drought.

PREMISE: Epiphytes are abundant in ecosystems such as tropical montane cloud forests where low-lying clouds are often in contact with vegetation. Climate projections for these regions include more variability in rainfall and an increase in cloud base heights, which would lead to drier conditions in the soil and atmosphere. While recent studies have examined the effects of drought on epiphytic water relations, the influence that atmospheric moisture has, either alone or in combination with drought, on the health and performance of epiphyte communities remains unclear. METHODS: We conducted a 10-week drought experiment on seven vascular epiphyte species in two shadehouses, one with warmer and drier conditions and another that was cooler and more humid. We measured water relations across control and drought-treatment groups and assessed functional traits of leaves produced during drought conditions to evaluate trait plasticity. RESULTS: Epiphytes exposed to drought and drier atmospheric conditions had a significant reduction in stomatal conductance and leaf water potential and an increase in leaf dry matter. Nonsucculent epiphytes from the drier shadehouse had the greatest shifts in functional traits, whereas succulent epiphytes released stored leaf water to maintain water status. CONCLUSIONS: Individuals in the drier shadehouse had a substantial reduction in performance, whereas drought-treated individuals that experienced cloud immersion displayed minimal changes in water status. Our results indicate that projected increases in the cloud base height will reduce growth and performance of epiphytic communities and that nonsucculent epiphytes may be particularly vulnerable.

Development of Behaviorally Based Monitoring and Biosurveillance Tools for the Invasive Spotted Lanternfly (Hemiptera: Fulgoridae).

The spotted lanternfly, Lycorma delicatula White, is an invasive planthopper (Hemiptera: Fulgoridae) that was first detected in the United States in Berks County, PA, in 2014, and has since spread in the mid-Atlantic region. This phloem-feeding pest has a broad host range, including economically important crops such as grape where their feeding causes dieback of infested plants. Monitoring the presence and abundance of L. delicatula is of utmost importance to develop pest management approaches. Current monitoring practices include sticky bands deployed on tree trunks, sometimes paired with commercially available methyl salicylate lures. A drawback associated with sticky bands is the high numbers of nontarget captures. Here, we developed traps for L. delicatula based on a circle trap originally designed for weevils. These traps are comprised of a screen funnel that wraps around the trunk of a tree and guides individuals walking up the trunk into a collection device. In 2018 and 2019, we compared circle trap designs with sticky bands in Pennsylvania and Virginia. In both years, circle trap designs yielded captures that were equivalent to or exceeded captures of L. delicatula on sticky bands. Nontarget captures were significantly lower for circle traps compared with sticky bands. Presence of a methyl salicylate lure in association with traps deployed on host trees or vertical tree-mimicking posts did not increase L. delicatula captures compared with unbaited traps. Circle traps, modified using vinyl screen and a larger collection device, present an alternative to the current approach with reduced nontarget capture for monitoring L. delicatula.

Patterns of Seismicity Associated with USGS Identified Areas of Potentially Induced Seismicity.

A systematic review across U.S. Geological Survey (USGS) identified potentially induced seismic locations was conducted to discover seismic distance patterns and trends over time away from injection disposal wells. Previous research indicates a 10 km (6 miles) average where the majority of induced seismicity is expected to occur within individual locations, with some areas reporting a larger radius of 35 km (22 miles) to over 70 km (43 miles). This research analyzed earthquake occurrences within nine USGS locations where specified wells were identified as contributors to induced seismicity to determine distance patterns from disposal wells or outward seismic migration over time using established principles of hydrogeology. Results indicate a radius of 31.6 km (20 miles) where 90% of felt earthquakes occur among locations, with the closest proximal felt seismic events, on average, occurring 3 km (1.9 miles) away from injection disposal wells. The results of this research found distance trends across multiple locations of potentially induced seismicity.

Cells in the female retrotrapezoid region upregulate c-fos in response to 10%, but not 5%, carbon dioxide.

The retrotrapezoid nucleus (RTN) is thought to regulate breathing in response to changes in blood carbon dioxide (CO(2)), and to make a vital contribution to respiratory drive, especially during sleep. However, cells in the female RTN fail to upregulate c-fos in response to low level CO(2) exposure, while cells in the male RTN have a robust upregulation of c-fos in response to low level CO(2) exposure. In this study, we examined the possibility that the female RTN has a higher threshold for c-fos upregulation in response to CO(2). Following exposure of Fos-Tau-LacZ (FTL) transgenic mice to 10% CO(2), c-fos was upregulated in just as many cells in the female as in the male RTN. In addition, the male RTN responded equivalently to 5% and 10% CO(2), consistent with a lack of a dose response to CO(2) in the male RTN. Cells in the nearby facial nucleus upregulated c-fos in the same number of cells regardless of sex or gas exposure, confirming that the sex difference in the RTN is unique to that nucleus. We propose that the male and female RTN upregulate c-fos differently in response to CO(2) due to differences in the transcriptional regulation by estrogens of genes that encode proteins related to neuronal excitability or specifically related to central chemoreception, such as potassium channels. These findings could have clinical relevance to sleep related breathing disorders that disproportionately affect males, including the sudden infant death syndrome and sleep apnea.