bluuebunny/crossref_metadata_2025 huggingface dataset:
PENERAPAN PERMAINAN “ SMART EGG PUZZLE ” DALAM MENINGKATKAN PERILAKU PROSOSIAL ANAK USIA DINI
Authors: Puspitasari Endang
Published: January 2022

The purpose of this study was to determine the effect of the application of the smart egg puzzle game on the social behavior of early childhood. This research was conducted in TK intan Berlian Lamongan with a total sample of 35 children. This study uses a quantitative approach. The research design in this study uses a quasi-experimental research design, where this design has a control group but does not fully function to control external variables that affect the implementation of the experiment. In this study, a nonequivalent control group design was used, where the experimental group and the control group were not chosen randomly. The indicators of this research are being cooperative, expressing emotions according to existing conditions and sharing with others. The results showed that the smart egg puzzle game had a significant effect on understanding the concept of geometry and social behavior in children aged 5-6 years at Intan Berlian Kindergarten. The t-test proves that the value of sig. Levene's Test for Equality of Variances is 0.221, greater than 0.05, it means that H0 is rejected and Ha is accepted. Thus it can be concluded that there is a significant difference between the experimental group and the control group
Sustained Suppression of Pythium Diseases: Interactions between Compost Maturity and Nutritional Requirements of Biocontrol Agents
Authors: Hoitink Harry A.J. , Hadar Yitzhak , Madden Laurence V. , Chen Yona
Published: January 2022

Several procedures were developed that predict maturity (stability) of composts prepared from municipal solid wastes (MSW). A respirometry procedure, based O2 uptake by compost, predicted (R2=0.90) the growth response of ryegrass in composts and an acceptable level of maturity. Spectroscopic methods (CPMAS13-NMR and DRIFT spectroscopy) showed that the stabilizing compost contained increasing levels of aromatic structures. All procedures predicted acceptable plant growth after approximately 110 days of composting. MSW compost suppressed diseases caused by a broad spectrum of plant pathogens including Rhizoctonia solani, Pythium aphanidermatum and Fusarium oxysporum. A strain of Pantoea agglomerans was identified that caused lysis of hyphae of R. solani. Evidence was obtained, suggesting that thermophilic biocontrol agents also might play a role in suppression. 13C-NMR spectra revealed that the longevity of the suppressive effect against Pythium root rot was determined by the concentration of readily biodegradable carbohydrate in the substrate, mostly present as cellulose. Bacterial species capable of inducing biocontrol were replaced by those not effective as suppression was lost. The rate of uptake of 14C-acetate into microbial biomass in the conducive substrate was not significantly different from that in the suppressive substrate although specific activity was higher. The suppressive composts induced systemic acquired resistance in cucumjber roots to Pythium root rot and to anthracnose in the foliage. Composts also increased peroxidase activity in plants by the conducive substrate did not have these effects. In summary, the composition of the organic fraction determined bacterial species composition and activity in the substrate, which in turn regulated plant gene expression relative to biological control.
Host Plant Kairomonal Effects on Heliothis Mate-Finding Behavior
Authors: Landolt Peter , Dunkelblum Ezra , Heath Robert , Kehat Moshe
Published: January 2022

We investigated the possible attraction of species of Helicoverpa and Heliothis to host plant odors and to male pheromone as a prelude to a larger investigation of host chemical effects on moth sexual and reproductive behavior. Both sexes of H. armigera and H. peltigera were attracted to particular host plants or host plant extracts. Heliothis virescens was also found to be attracted to a flowering shrub it may use as a nectar source. Unmated females of H. armigera and H. peltigera were attracted to live males and to a male pheromone extract prepared from exposed filter papers in cages of males. Some attraction of unmated female H. zea to males was also evident, but at much lower response rate.
The Role of Phytohormones in the Response of Plants to Salinity Stress
Authors: Bray Elizabeth , Lerner Zvi , Poljakoff-Mayber Alexander
Published: January 2022

Salinity is an increasing problem in many irrigated areas of crop production and is a significant factor in reducing crop productivity. Developmental, physiological, and molecular responses to salinity were studied in order to improve our understanding of these responses. Improvements in our understanding of plant responses to salinity are necessary in order to develop crops with improved salt tolerance. Previously, in Israel, it was shown that Sorghum biccolor can adapt to an otherwise lethal concentration of NaCl. These experiments were refined and it was shown that there is a specific window of development in which this adaption can occur. Past the window of development, Sorghum plants can not be adapted. In addition, the ability to adapt is not present in all genotypes of Sorghum. Cultivars that adapt have an increased coefficient of variation for many of the physiological parameters measured during the mid-phase of adaptation. Therefore, it is possible that the adaptation process does not occur identically in the entire population. A novel gene was identified, isolated and characterized from Sorghum that is induced in roots in response to salinity. This gene is expressed in roots in response to salt treatments, but it is not salt-induced in leaves. In leaves, the gene is expressed without a salt treatment. The gene encodes a proline-rich protein with a novel proline repeat, PEPK, repeated more than 50 times. An antibody produced to the PEPK repeat was used to show that the PEPK protein is present in the endodermal cell wall of the root during salt treatments. In the leaves, the protein is also found predominantly in the cell wall and is present mainly in the mesophyll cells. It is proposed that this protein is involved in the maintenance of solute concentration.
Calcium-Mediated Signal Transduction during Leaf Senescence
Authors: Philosoph-Hadas Sonia , Crain Richard , Meir Shimon , Aharoni Nehemia , Lurie Susan
Published: January 2022

We have examined the possibility that modulation of [Ca2+]cyt may represent a signal which induces senescence processes in leaves, through triggering of lipid hydrolysis leading to the cascade of detriorative events. Characterization of the signal transduction components operating during leaf senescence was gained by studying various Ca2+-dependent activities of parsley and chrysanthemum leaves, in relation to several senescence functions, and in response to senescence-modulating hormones (ethylene,ABA, BA and IAA). Some innovative findings regarding the control of senescence processes by [Ca2+]cyt were established: Several Ca2+-or CaM-related compounds were shown to modulate [Ca2+]cyt and action, thereby affecting whole leaf senescence. The involvement of [Ca2+]cyt in mediating the effects of senescence-modulating hormones has been demonstrated. Loss of energized Ca2+-transport capability of PM was found to an early event in leaf senescence, which occurs before changes in senescence parameters are observed, and while other PM ATPase enzymes still retain about 50% of their activities. A general pattern of increased phosphorylation of PM proteins with advanced senescence, which could be modified by plant hormones applied in vivo (BA) or in vitro (ABA), sa found. Taken together, all this indirect evidence indicate that [Ca2+]cyt is elevated due to the senescence-induced decrease in the ability to extrude Ca2+, which results particularly from reduced PM Ca2++-transport capability rather than increased operation of Ca2+ channels or elevated Ins(1,4,5)P3 levels. The direct proof for such a senescence-related elevation in [Ca2+]cyt was provided for the first time by the Ca2+ imaging measures with fura-2, showing a rise in [Ca2+]cyt of mesophyll cells upon senescence induction, which preceeded changes in typical senescence characteristics. This research provides strong evidence for regarding the rise in [Ca2+]cyt as a primary event in induction of the senescence syndrome in detached leaves. The findings have also broad implications for postharvest handling of leafy crops and ornamentals, and open new avenues for employing Ca2+-related inhibitors to delay leaf senescence.
Role of Molecular Chaperone in Wheat Storage Protein Assembly
Authors: Galili Gad , Bennett Alan
Published: January 2022

Following sequestration into the ER, wheat gliadins assemble into complexes that initiate the formation of protein bodies. In the present work we have characterized the DNA sequence and regulation of expression of a plant BiP and also studied its interaction with wheat storage proteins as well as its role in the maturation of these storage proteins. In the Israeli lab, immunoprecipitation studies were made using anti BiP and anti storage proteins sera, both in wheat and in transgenic tobacco plants expressing a wheat gliadin storage proteins. In both cases, we could show that BiP interacts with the gliadin storage proteins. In addition, we could show that BiP also played an important role in the initial assembly of the gliadins. In the American lab, the complexity, structure and properties of tomato BiP was characterized at the molecular and biochemical levels. In addition, tomato BiP was also overexpressed in bacteria and the overexpressed protein was found to be active. The cooperative findings of the Israeli and American labs clearly improves our understanding of the structure and expression of a plant BiP as well as its role in the maturation of storage proteins in plants seeds. In addition, it will serve as a foundation for future studies of the mechanisms of BiP function in in vitro studies using purified storage proteins and purified recombinant active BiP.
Cloning a Fusarium Resistance Gene in Tomato Based on Knowledge of its Map Position
Authors: Zamir Dani , Tanksley Steven , Fluhr Robert
Published: January 2022

The objectives of this project were to develop the tools and methodologies for positional cloning of genes in tomato and apply them for the cloning a Fusarium resistance gene - I2.. The feasibility of positional cloning of disease resistance genes was demonstrated for Pto which confers resistance to pseudomonas (Martin et al. 1993). The Fusarium resistance gene was mapped genetically and physically and was found to be in close proximity to TG 105 (Segal et al. 1992). To obtain fine mapping of gene I2, and additional target genes in future projects, a high density linkage map was developed (Tanksley et al. 1992; Broun and Tanksley 1993). In addition two permanent mapping populations were constructed: a recombinant inbred (Paran et al. 1995; Zamir et al. 1993) and an introgression line population (Eshed et al. 1992; Eshed and Zamir 1994). Using these resources we determined that the I2 locus shows complete co-segregation, down to a resolution of a few Kb, with SL8 which shows architectural similarity with other plant resistance genes. Transformation and complementation analysis is in progress (Ori et al. in preparation).
Development and Evaluation of a Method of Hormonal Treatment to Increase Fertility in Dairy Cows
Authors: Britt Jack , Rosenberg Miriam , Washburn Steven , Kaim Moshe
Published: January 2022

The objectives were: 1) to develop a practical method for improving conception rates of dairy cows by administering progesterone during the critical period of the luteal phase prior to first insemination, 2) to determine which cows respond best to such a progesterone treatment, and 3) to determine physiological changes that are associated with this treatment. Experiment 1, conducted in the US and Israel, tested three treatment protocols. Holstein cows: 1) served as controls, 2) received two injections of prostaglanding F2a (PGF) 14 days apart, or 3) received two PGF injections and were inserted with a progesterone releasing intravaginal device (PRID) for 6 days beginning 9 days after the first injection of PGF. Fertility was positively related to plasma progesterone concentations prior to the second PGF injection, but PRID treatment did not improve fertility in cows with low progesterone. Cows in synchronized estrus 2-8 days after the second injection of PGF had higher fertility and better reproductive performance than non-synchronized cows. Experiment 3, conducted in Israel, involved three studies that assessed effect of PRID treatment on: 1) progesterone concentration in ovarian tissue and uterine tissue and fluids, 2) changes in systemic LH and estradiol, and 3) development and function of dominant follicles. Treatment with PRID increased progesterone in uterine tissue and fluid exclusive of effects on systemic concentrations. Low progesterone resulted in persistent follicles that exceeded normal size and produced excessive estrogen. Experiment 4, conducted in the US, evaluated influence of endogenous progesterone on embryo development. Cows were given two injections of PGF 14 days apart and single embryos were recovered 7 days after estrus. Among cyclic cows, there was a linear relationship between progesterone concentration and percentage of cows from which an embryo was recovered. Overall these experiments show that systemic progesterone concentrations affect fertility in high-producing dairy cows through effects on follicular development and embryo survival
Rapid, Sensitive Bluetongue Virus Serogroup and Serotype Detection Using Polymerase Chain Reaction
Authors: Osburn Bennie , Ianconescu Marius , Akita Geoffrey , Kaufman Rozalia
Published: January 2022

The objectives of this proposal were to enhance animal health by 1) development of a BTV serogroup diagnostic assay using polymerase chain reaction (PCR) and 2) development of a BTV serotype specific diagnostic PCR assay. A PCR assay for diagnosis of bluetongue virus (BTV) serogroup from clinical samples meeting the criteria of objective 1 was developed. This PCR assay is more sensitive than virus isolation and has been adopted by both the U.S. and Israeli collaborating laboratories of this project, as well as at least one other U.S. laboratory for routine diagnosis of BTV infection in ruminants. The basic BTV PCR protocol has also become an essential tool in BTV molecular research in both collaborating laboratories. During development of the BTV serotype specific PCR we had the opportunity to investigate a nationwide outbreak of abortions and fatal disease in dogs in the U.S. purportedly due to BTV infection via a BTV contaminated canine vaccine. The BTV serogroup PCR was integral in confirming BTV in tissues from affected dogs and in lots of the suspect vaccine. This led to the first published report of BTV infection in dogs. We discovered that BTV can produce silent persistent infection in canine cell culture. This indicated a need for more stringent screening of biologics for occult BTV infection. A novel mixed cell culture method was developed to identify occult BTV and other occult viral infection cell cultures. Serotype specific primers for PCR detection of all U.S. BTV serotypes and two Israel serotypes (BTV-2 and 10) have been evaluated and are available. A subsequent collaboration would logically include sequencing of the L2 genes of Israel BTV-4, 6 and 16, allowing incorporation of these Israel BTV serotypes into a multiplex PCR assay.
Genetic Analysis in vitis Using Molecular Markers
Authors: Reisch Bruce , Spiegel-Roy Pinhas , Weeden Norman , Ben-Hayyim Gozal , Beckmann Jacques
Published: January 2022

Genetic analysis and mapping in grapes has been difficult because of the long generation period and paucity of genetic markers. In the present work, chromosome linkage maps were developed with RAPD, RFLP and isozyme loci in interspecific hybrid cultivars, and RAPD markers were produced in a V. vinifera population. In three cultivars, there were 19 linkage groups as expected for a species with 38 somatic chromosomes. These maps were used to locate chromosome regions with linkages to important genes, including those influencing powdery mildew and botrytis bunch rot resistance; flower sex; and berry shape. In V. vinifera, the occurrence of specific markers was correlated with seedlessness, muscat flavor and fruit color. Polymorphic RAPD bands included single copy as well as repetitive DNA. Mapping procedures were improved by optimizing PCR parameters with grape DNA; by the development of an efficient DNA extraction protocol; and with the use of long (17- to 24-mer) primers which amplify more polymorphic loci per primer. DNA fingerprint analysis with RAPD markers indicated that vinifera cultivars could be separated readily with RAPD profiles. Pinot gris, thought to be a sort of Pinot noir, differed by 12 bands from Pinot noir. This suggests that while Pinot gris may be related to Pinot noir, it is not likely to be a clone. The techniques developed in this project are now being further refined to use marker-assisted selection in breeding programs for the early selection of elite seedlings. Furthermore, the stage has been set for future attempts to clone genes from grapes based upon map locations.
Production of Antibodies to Citrus Tristeza Virus in Transgenic Citrus
Authors: Lee Richard , Bar-Joseph Moshe , Derrick K.S. , Vardi Aliza , Brlansky Roland , Eshdat Yuval , Powell Charles
Published: January 2022

Citrus tristeza virus (CTV) is the most important virus disease of citrus in the world. CTV causes death of trees on sour orange rootstock and/or stem pitting of scions regardless of rootstock which results in trees of low vigor, reduced yield with reduction in size and quality of fruit. The purpose of this project was to produce monoclonal antibodies (MABs) to CTV coat protein (CP), develop single domain antibodies (dAbs) or Fab fragments which neutralize the infection by binding to the virus, and to produce transformed plants which express the dAbs. The objectives of this research have been met and putative transgenic tobacco and citrus plants have been developed. These putative transgenic plants are presently undergoing evaluation to determine the level of dAbs expression and to determine their resistance to CTV. Additionally, the CTV genome has been sequenced and the CP gene of several biologically characterized CTV strains molecular characterized. This has indicated a correlation between CP sequence homology and biological activity, and the finding of DI RNAs associated with some CTV strains. Several MABs have been produced which enable broad spectrum identification of CTV strains while other MABs enable differentiation between mild and severe strains. The use of selected MAbs and determination of the CP gene sequence has enabled predictions of biological activities of unknown CTV isolates. The epitopes of two MABs, one reacting selectively with severe CTV strains and the other reacting with all strains, have been characterized at the molecular level.
Asymmetric Somatic Hybridization: Developing a Gene Transfer System for Solanaceous Vegetable Crops
Authors: Sink Ken , Izhar Shamay , Nachmias Abraham
Published: January 2022

Highly asymmetric somatic hybrid plants were obtained by PEG/DMSO fusion of gamma irradiated (100, 250, 7500 and 1000 Gy) protoplasts of a (KmR-) interspecific hybrid Lycopersicon esculentum x L. pennellii (EP) with protoplasts of eggplant (E). Somatic hybrid calli were selected based on kanamycin resistance and verified by PCR of the NptII gene, RAPD's and Southern's using potato rDNA pTHG2 probes. Flow cytometry indicated all hybrid calli that did not regenerate shoots were 5-9n. Three asymmetric plants regenerated only from callus close to 4n and such calli oly occurred when EP received 100 Gy. The asymmetric plants had eggplant morphology and regenerated from one hybrid callus with 6.29 average size tomato chromosomes. Limited amounts of EP DNA were found in the three somatic hybrid plants H18-1 to -3 by dot-blot hybridization with probe pTHG2, to be equivalent to 6.23, 5.41, and 5.95 % EP, respectively. RFLP analysis of Lycopersicon esculentum and L. pennellii specific chromosomes revealed that only fragments of 8 to 10 out of the 24 EP chromosomes are present in the asymmetric plants. Transgenic plants 2-3, 2-4 and 10-3 were found resistant to verticillium; suggesting successful transfer of the Ve complex from S. torvum to eggplant.
Mode of Action of Yeast Biocontrol Agents of Postharvest Diseases of Fruits
Authors: Chalutz Edo , Wisniewski Michael , Droby Samir , Eilam Yael , Chet Ilan
Published: January 2022

In a previous BARD-supported study, three of the investigators of this research were involved in a study on biological control of postharvest diseases of citrus and deciduous fruits. Several naturally occurring, non-antibiotic producing yeast antagonists were identified. Application of some of these antagonists resulted in very high levels of biocontrol under laboratory conditions but lower efficacy in semi-commercial tests. It was felt that the lack of knowledge on the mode of action of the biocontrol agents was limiting their efficient use. The current study was aimed at narrowing this gap in our knowledge. Two specific objectives were outlined: to study the mechanism by which calcium salts enhance biocontrol activity and to determine the role, if any, of the yeast extracellular materials and/or enzymes which degrade fungal cell walls during the interaction between the antagonists, the pathogen and the host. CaCl2 but not MgCl2, inhibited spore germination, and germ-tube elongation of Botrytis cinerea, Penicillium expansum and P. digitatum in culture. It also inhibited the pectinolytic activity of the pathogens. Biocontrol of apple decay by isolate 182 of Candida oleophila, an effective biocontrol agent, was enhanced by the addition of CaCl2 whereas there was no effect on the biocontrol activity of isolate 247 of this yeast. Similarly, CaCl2 enhanced efficacy of the US-7 isolate of Pichia guilliermondii in reducing infection of P. digitatum in citrus fruit. CaCl2 by itself also reduced the infection of peel wounds and stimulated ethylene production by grapefruit peel. This antagonist exhibited a very high ability to maintain cytosolic Ca2+ homeostasis when exposed to high CaCl2 concentrations. It is postulated, therefore, that enhanced biocontrol activity by calcium is the result of direct inhibition of the pathogen by calcium ions on spore germination and metabolism and indirectly due to the ability of the biocontrol agent to maintain normal metabolism in the presence of high levels of calcium. The extracellular materials produced by P. guilliermondii in culture and on the fruit inhibited, at low concentrations, the pathogen in culture and reduced percent infection of the fruit. The direct inhibition of the pathogen by these materials may thus be involved in the mode of action of the antagonist. This study contributed to our knowledge on the action of calcium salts and the yeast antagonist extracellular materials on biocontrol activity and will contribute to a more efficient use of this technology in the control of postharvest diseases of fruits.
Reduction of Reticuloendotheliosis in Foundation Breeding Flocks of Chickens: A Combined Immunological and Molecular Biological Approach
Authors: Malkinson Mertyn , Witter Richard , Davidson Irit
Published: January 2022

Reticuloendotheliosis virus (REV) is an avian retrovirus that can cause immunosuppression, growth retardation and tumors. An attempt to define the extent of the economic damage to the poultry industry that it causes is discussed in this report. In addition to losses experienced by commercial laying flocks, reduced rates of hatchability and embryo developmental disorders were demonstrable due to vertical transmission of the virus. I. Eradication of REV In this project a comprehensive national program was applied for the eradication of REV from Israeli breeding stocks by the elimination of antibody-positive birds from the breeding program. The prevention of REV-infected breeders entering Israel was also implemented by serological examination of imported day-old chickens and turkeys for maternal antibody. At the same time commercial breeding flocks in Israel were surveyed routinely to measure the extent of environmental exposure to REV throughout Israel. II. Economic factors associated with vertical transmission on breeders and progeny It was observed that on some poultry farms exposure of breeding flocks to viral infection, if it occurs when the birds are immunocompetent, leads only to a seroconvertion event. In these flocks no differences were demonstrated between the performances of seronegative and seropositive birds. When the F1 generation was selected according to seronegativity of the parents, all the progeny were seronegative, indicating that tolerantly infected birds did not form a significant proportion of the parent flock. In sharp contrast, breeding flocks that became exposed to the virus about the point of lay or during the laying period, shed virus vertically for a brief period of time through the egg. Our epizootiological observations lead us to conclude that the progeny (laying pullets) becomes tolerantly infected and are immunosuppressed as they increase in age. Increased mortality and susceptibility to intercurrent diseases were recorded.
Decision Making within Plant Root Systems
Authors: Waisel Yoav , McMichael Bobbie , Eshel Amram
Published: January 2022

Architecture of a root system is the expression of the potential of various root types to branch, to grow and to coordinate with other plant organs, under the specific limitations of the environmental conditions. The present investigation has proven the following points. 1) Genotypes with different types of root systems were identified. The growth patterns of their roots and the distribution of laterals along their main axes were recorded. 2) The patterns of development of the root systems of four cotton genotypes, throughout the entire life cycle of the plants, were described, even at such a late stage of development when the total length of the roots exceeded two kilometers. To the best of our knowledge, this is the first time that an analysis of this type is accomplished. 3) The development of root systems under restrictive soil conditions were compared with those that have developed under the non-restrictive conditions of aeroponics. Results indicate that in the absence of the mechanical impedance of the soil, cotton plants develop single roots that reach the length of 6 m, and have a total root length of 2000 m. Thus, root growth is strongly inhibited by the soil, with some root types being inhibited more than others. 4) One of the important decisions, in constructing an operational root system architecture of mature plants, is the shift of the balance between various root fractions in favor of the very fine roots. 5) Root system architecture is determined, in part, by the sites of initiation of the lateral roots. This is determined genetically by the number of xylem archs and by the totuosity of the stele. Selection for such traits should be sought.
Endophytic Bacteria of Cotton and Sweet Corn for Providing Growth Promotion and Biological Disease Control
Authors: Kloepper Joseph W. , Chet Ilan
Published: January 2022

Endophytes were isolated from 16.7% of surface-disinfested seeds and 100% of stems and roots of field-growth plants. Strains from Israel with broad-spectrum in vitro antibiosis were mainly Bacillus spp., and some were chitinolytic. Following dipping of cut cotton roots into suspensions of these strains, endophytes were detected up to 72 days later by isolation and by autoradiograms of 14C-labelled bacteria. Selected endophytes exhibited biological control potential based on significant reductions in disease severity on cotton inoculated with Rhizoctonia solani or Fusarium oxysporum f. sp. vasinfectum as well as control of Sclerotium rolfsii on bean. Neither salicylic acid nor chitinase levels increased in plants as a result of endophytic colonization, suggesting that the observed biocontrol was not accounted for by PR protein production. Some biocontrol endophytes secreted chitinolytic enzymes. Model endophytic strains inoculated into cotton stems via stem injection showed only limited movement within the stem. When introduced into stems at low concentrations, endophytes increased in population density at the injection site. After examining several experimental and semi-practical inoculation systems, seed treatment was selected as an efficient way to reintroduce most endophytes into plants.
Novel streaming potential and thermal sensor techniques for monitoring water and nutrient fluxes in the vadose zone
Authors: Jones Scott B. , Friedman Shmuel P. , Communar Gregory
Published: January 2022

The “Novel streaming potential (SP) and thermal sensor techniques for monitoring water and nutrient fluxes in the vadose zone” project ended Oct. 30, 2015, after an extension to complete travel and intellectual exchange of ideas and sensors. A significant component of this project was the development and testing of the Penta-needle Heat Pulse Probe (PHPP) in addition to testing of the streaming potential concept, both aimed at soil water flux determination. The PHPP was successfully completed and shown to provide soil water flux estimates down to 1 cm day⁻¹ with altered heat input and timing as well as use of larger heater needles. The PHPP was developed by Scott B. Jones at Utah State University with a plan to share sensors with Shmulik P. Friedman, the ARO collaborator. Delays in completion of the PHPP resulted in limited testing at USU and a late delivery of sensors (Sept. 2015) to Dr. Friedman. Two key aspects of the subsurface water flux sensor development that delayed the availability of the PHPP sensors were the addition of integrated electrical conductivity measurements (available in February 2015) and resolution of bugs in the microcontroller firmware (problems resolved in April 2015). Furthermore, testing of the streaming potential method with a wide variety of non-polarizable electrodes at both institutions was not successful as a practical measurement tool for water flux due to numerous sources of interference and the M.S. student in Israel terminated his program prematurely for personal reasons. In spite of these challenges, the project funded several undergraduate students building sensors and several master’s students and postdocs participating in theory and sensor development and testing. Four peer-reviewed journal articles have been published or submitted to date and six oral/poster presentations were also delivered by various authors associated with this project. We intend to continue testing the "new generation" PHPP probes at both USU and at the ARO resulting in several additional publications coming from this follow-on research. Furthermore, Jones is presently awaiting word on an internal grant application for commercialization of the PHPP at USU.
Antimicrobial and antioxidant functionalized nanoparticles for enhancing food safety and quality: proof of concept
Authors: Choudhary Ruplal , Rodov Victor , Kohli Punit , Haddock John D. , Droby Samir
Published: January 2022

General concept. The reported 1-year study tested the feasibility ofpreparing antimicrobial and antioxidant nanoparticlesfunctionalized with natural phenolic compounds, as a first step to reach the ultimate goal - improving safely and quality of foods by developing novel antimicrobial and antioxidant food-contacting materials. The secondary objectives of the study were (a) selecting the most promising phenoliccompounds, (b) building nanoparticles with the selected phenolicgrafted on their Surface, and (c) testing antimicrobial and antioxidant properties of these particles. The study was expected to provide a " go/no go" decision as concerning the prospects of phenolic- bound nanoparticles as antimicrobial and antioxidant agents. Results. In course of the feasibility study, curucminwas chosen as the most promising phenoliccompound due to its high antibacterial activity exceeding other tested compounds by at leas one order of magnitude. Lipsome-typephospholipid/polydiacetylene(PDA) nanoparticlesfunctionalizedwith curcuminwere successfully built. The pitfall of limited curcumin amount that could be covalently bound to theparticle surface was circumvented by inclusion of curcunun in the liposome body. It was suggested onthe basis of fluorescence spectroscopy that curcuminwas bound by hydrophobic forces in the bi1ayer periphery of the Liposomesand therefore mightexert a contact effect on microorganisms. The curcumin­ functionalizednanoparticles(CFN) were shown to have a strong bactericidal activity towards both Gram-negative (E. coli) and Gram-positive (B. ce,·e11s) bacteria, but only limited effect against yeast. Furthermore, beyond the originallyplanned objectives, preliminary trials showed that CFN could be bound to silanized glass surface rendering aנבtiנnicrobial activity to the glass. Tnaddition, the particles showed antioxidantcapacity. Tberefore, it ,vas co11cluded tlוattlוeaims of tlוefeasibility study bad been successfully reached an
Elucidating the impact of enhanced conversion of primary to secondary metabolism on phenylpropanoids secondary metabolites associated with flavor, aroma and health in tomato fruits
Authors: Galili Gad , Klee Harry J. , Aharoni Asaph
Published: January 2022

• Targeted manipulating Phenylalanine (Phe) synthesis is one of the most powerful strategies to boost the biologically and economically important secondary metabolites, including phenylpropaniods, aromatic volatiles and specialized secondary metabolites. • Over-expression of the petunia MYB transcript factor, ODORANT1 (ODO1), results in significant alterations of the levels of specific phenylpropanoid compounds in plants. • Our previous studies indicated that ectopic expression of the feedback-insensitive AroG could break the bottleneck between primary and secondary metabolisms in tomato, thereby aiding in producing new tomato composition and identifying the unknown roles of multiple key regulators in specialized metabolism. Therefore, combining the AroG and ODO1 is of particular interest for elucidating the combined regulatory role of both of these genes in the Phe metabolic pathway, as well as generating tomato fruits that contain higher levels of secondary metabolites. • Here, we performed the LC-MS and GC-MS analyses on fruits of four tomato genotypes, namely, wild type tomato fruits as well as tomato fruits expressing the AroG, ODO1 and the combination of AroG plus ODO1 (AO) genotypes. Our results elaborated that the levels of many of the Phe-derived metabolites were predominately altered in fruits of the AO genotype, compared to tomato fruits expressing either AroG or ODO1 individually. The levels of most of these metabolites were significantly stimulated, such as Tyrosine (Tyr), coumaric acid and ferulic acid derived metabolites, but the levels of some important secondary metabolites were reduced in the AO transgenic genotypes as compared to either AroG or ODO1 lines. Nevertheless, our results also revealed that the levels of aromatic volatiles were obviously down regulated in the AO, compared to that in AroG transgenic fruits, but were boosted while compared to the wild type and ODO1 transgenic fruits. • Our results suggest that ODO1 expression may also have a negative effect on the production of some of the aromatic volatiles in tomato fruits, indicating that ODO1 acts as an important regulator of the shikimate pathway, which leads to the production of the aromatic amino acids and secondary metabolites derived from them. Key words: AroG, ODO1, tomato, metabolism, shikimate pathway
Specific mediators of auxin activity during tomato leaf and fruit development
Authors: Ori Naomi , Estelle Mark
Published: January 2022

The plant hormone auxin is involved in numerous developmental processes, including leaf and fruit development. The tomato (Solanumlycopersicum) gene ENTIRE (E) encodes an auxin-response inhibitor from the Aux/IAA family. While most loss-offunction mutations in Aux/IAA genes are similar to the wild type due to genetic redundancy, entire (e) mutants show specific effects on leaf and fruit development. e mutants have simple leaves, in contrast to the compound leaves of wild type tomatoes. In addition, e plants produce parthenocarpic fruits, in which fruit set occurs independently of fertilization. The aim of this research program was to utilize the e mutation to identify and characterize genes that mediate the specific effect of auxin in leaf and fruit development. The specific objectives of the project were to: 1. Characterize and map modifiers of the e leaf phenotype. 2. Characterize and map suppressors of the e fruit phenotype. 3. Dissect the developmental specificity of the E gene. 4. Examine the effect of fruit-overexpression of identified genes on fruit set and seed production. To identify mediators of auxin in leaf development, we mainly focused on one mutant, crawling elephant (crel, previously called t282), which showed substantial suppression of the e phenotype and other auxin-relatedphenotypes. We have identified the CREL gene as a homolog of the Arabidopsis VRN5 gene, involved in recruiting polycomb silencing complexes to specific targets. We showed that CREL affects auxin sensitivity in tomato. Suppressors of the e fruit phenotype have been further characterized and selected for more profound effects. Expression profiling by RNAseq was used to analyze the effect of e as well as crel on gene expression in leaves and fruits. This analysis has identified putative E and CREL targets. We have initiated studies to assess the role of some of these targets in flower and fruit development. The research has identified potential mediators of auxin response in leaf, flower and fruit development.
Novel regulation of transpiration by sugar signals within guard cells
Authors: Granot David , Assmann Sarah M.
Published: January 2022

Water is the major limiting factor in agriculture and stomata, composed of two guard cells and the pore they circumscribe, are the chief gates controlling plants’ water loss. The prevailing century old paradigm was that sugars act as an osmoticum in guard cells, contributing to the opening of the stomata. In contrast, we discovered that sugars close stomata and the closure is mediated by the sugar-sensing enzyme hexokinase (HXK) that triggers the abscisic acid (ABA)-signaling pathway within the guard cells. This new discovery suggests a sugar-sensing mechanism within guard cells that controls stomatal closure, and supports the existence of a stomatal feedback mechanism that coordinates photosynthesis with transpiration.
Renewable, long-term, antimicrobial surface treatments through dopamine-mediated binding of peptidoglycan hydrolases
Authors: Rahimipour Shai , Donovan David
Published: January 2022

There is a need for renewable antimicrobial surface treatments that are semi- permanent, can eradicate both biofilms and planktonic pathogens over long periods of time and that do not select for resistant strains. This proposal describes a dopamine binding technology that is inexpensive, bio-friendly, non-toxic, and uses straight-forward commercially available products. The antimicrobial agents are peptidoglycanhydrolase enzymes that are non-toxic and highly refractory to resistance development. The goal of this project is to create a treatment that will be applicable to a wide variety of surfaces and will convey long-lasting antimicrobial activity. Although the immediate goal is to create staphylolytic surfaces, the technology should be applicable to any pathogen and will thus contribute to no less than 3 BARD priorities: 1) increased animal production by protecting animals from invasive and emerging diseases, 2) Antimicrobial food packaging will improve food safety and security and 3) sustainable bio- energy systems will be supported by coating fermentation vats with antimicrobials that could protect ethanolic fermentations from Lactobacillus contamination that reduces ethanol yields. The dopamine-based modification of surfaces is inspired by the strong adhesion of mussel adhesion proteins to virtually all types of surfaces, including metals, polymers, and inorganic materials. Peptidoglycanhydrolases (PGHs) meet the criteria of a surface bound antimicrobial with their site of action being extracellular peptidoglycan (the structural basis of the bacterial cell wall) that when breached causes osmotic lysis. As a proof of principle, we will develop technology using peptidoglycanhydrolase enzymes that target Staphylococcus aureus, a notoriously contagious and antimicrobial-resistant pathogen. We will test for susceptibility of the coating to a variety of environmental stresses including UV light, abrasive cleaning and dessication. In order to avoid resistance development, we intend to use three unique, synergistic, simultaneous staphylococcal enzyme activities. The hydrolases are modular such that we have created fusion proteins with three lytic activities that are highly refractory to resistance development. It is essential to use multiple simultaneous activities to avoid selecting for antimicrobial resistant strains. This strategy is applicable to both Gram positive and negative pathogens. We anticipate that upon completion of this award the technology will be available for commercialization within the time required to achieve a suitable high volume production scheme for the required enzymes (~1-2 years). We expect the modified surface will remain antimicrobial for several days, and when necessary, the protocol for renewal of the surface will be easily applied in a diverse array of environments, from food processing plants to barnyards.
Dual role of the TYLCV protein V2 in suppressing the host plant defense
Authors: Gafni Yedidya , Lapidot Moshe , Citovsky Vitaly
Published: January 2022

TYLCV-Is is a major tomato pathogen, causing extensive crop losses in Israel and the U.S. We have identified a TYLCV-Is protein, V2, which acts as a suppressor of RNA silencing. Intriguingly, the counter-defense function of V2 may not be limited to silencing suppression. Our recent data suggest that V2 interacts with the tomato CYP1 protease. CYP1 belongs to the family of papain-like cysteine proteases which participate in programmed cell death (PCD) involved in plant defense against pathogens. Based on these data we proposed a model for dual action of V2 in suppressing the host antiviral defense: V2 targets SGS3 for degradation and V2 inhibits CYP1 activity. To study this we proposed to tackle three specific objectives. I. Characterize the role of V2 in SGS3 proteasomal degradation ubiquitination, II. Study the effects of V2 on CYP1 maturation, enzymatic activity, and accumulation and, III. Analyze the effects of the CYP1-V2 interaction on TYLCV-Is infection. Here we describe results from our study that support our hypothesis: the involvement of the host's innate immune system—in this case, PCD—in plant defense against TYLCV-Is. Also, we use TYLCV-Is to discover the molecular pathway(s) by which this plant virus counters this defense. Towards the end of our study we discovered an interesting involvement of the C2 protein encoded by TYLCV-Is in inducing Hypersensitive Response in N. benthamianaplants which is not the case when the whole viral genome is introduced. This might lead to a better understanding of the multiple processes involved in the way TYLCV is overcoming the defense mechanisms of the host plant cell. In a parallel research supporting the main goal described, we also investigated Agrobacteriumtumefaciens-encoded F-box protein VirF. It has been proposed that VirF targets a host protein for the UPS-mediated degradation, very much the way TYLCV V2 does. In our study, we identified one such interactor, an Arabidopsistrihelix-domain transcription factor VFP3, and further show that its very close homolog VFP5 also interacted with VirF. Interestingly, interactions of VirF with either VFP3 or VFP5 did not activate the host UPS, suggesting that VirF might play other UPS-independent roles in bacterial infection. Another target for VirF is VFP4, a transcription factor that both VirF and its plant functional homolog VBF target to degradation by UPS. Using RNA-seqtranscriptome analysis we showed that VFP4 regulates numerous plant genes involved in disease response, including responses to viral and bacterial infections. Detailed analyses of some of these genes indicated their involvement in plant protection against Agrobacterium infection. Thus, Agrobacterium may facilitate its infection by utilizing the host cell UPS to destabilize transcriptional regulators of the host disease response machinery that limits the infection.
Photoperiod effects on milk production in goats: Are they mediated by the molecular clock in the mammary gland?
Authors: Casey Therese , Mabjeesh Sameer J. , Shamay Avi , Plaut Karen
Published: January 2022

US scientists, Dr. Theresa Casey and Dr. Karen Plaut, collaborated with Israeli scientists, Dr. SameerMabjeesh and Dr. AviShamay to conduct studies proposed in the BARD Project No. US-4715-14 Photoperiod effects on milk production in goats: Are they mediated by the molecular clock in the mammary gland over the last 3 years. CLOCK and BMAL1 are core components of the circadian clock and as heterodimers function as a transcription factor to drive circadian-rhythms of gene expression. Studies of CLOCK-mutant mice found impaired mammary development in late pregnancy was related to poor lactation performance post-partum. To gain a better understanding of role of clock in regulation of mammary development studies were conducted with the mammary epithelial cell line HC11. Decreasing CLOCK protein levels using shRNA resulted in increased mammary epithelial cell growth rate and impaired differentiation, with lower expression of differentiation markers including ad herens junction protein and fatty acid synthesis genes. When BMAL1 was knocked out using CRISPR-CAS mammary epithelial cells had greater growth rate, but reached stationary phase at a lower density, with FACS indicating cells were growing and dying at a faster rate. Beta-casein milk protein levels were significantly decreased in BMAL1 knockout cells. ChIP-seq analysis was conducted to identify BMAL1 target genes in mammary epithelial cells. Studies conducted in goats found that photoperiod duration and physiological state affected the dynamics of the mammary clock. Effects were likely independent of the photoperiod effects on prolactin levels. Interestingly, circadian rhythms of core body temperature, which functions as a key synchronizing cue sent out by the central clock in the hypothalamus, were profoundly affected by photoperiod and physiological state. Data support that the clock in the mammary gland regulates genes important to development of the gland and milk synthesis. We also found the clock in the mammary is responsive to changes in physiological state and photoperiod, and thus may serve as a mechanism to establish milk production levels in response to environmental cues.