Capturing High Resolution Plant Movement in the Field.

Lodging of small grains due to environmental stresses results in yield loss, quality reduction, and difficulties with mechanical harvesting, which lead to economic consequences. New technological discoveries allow for faster and in situ measurements for determining the mechanics of loading stress and plant movement. The overall measurement of plant movement can be a very sophisticated method to mechanically test and predict the behavior of stems when exposed to wind. We investigated the inertial measurement of plants during different magnitude wind events. This type of analysis captures real time quantitative stem behavior during wind events. Using a 1.5 cm2 inertial measurement sensor attached to the upper panicle of a plant, we recorded the ranges and extremes of instantaneous linear acceleration and rotational velocity. When this technology was applied to historically known varieties of different lodging classification, the measurements were able to distinguish between cereal species and differences between movement of lodging susceptible and resistant plants without physical lodging. This type of technology could be used to improve field based lodging models and quantify movement resulting from micro changes in structural and composition of the stem, and to analyze plant movement in natural conditions with a resolution and specificity that has so far been prohibitively expensive and technologically challenging to achieve.

Image analysis and artificial intelligence in infectious disease diagnostics.

BACKGROUND: Microbiologists are valued for their time-honed skills in image analysis, including identification of pathogens and inflammatory context in Gram stains, ova and parasite preparations, blood smears and histopathologic slides. They also must classify colony growth on a variety of agar plates for triage and assessment. Recent advances in image analysis, in particular application of artificial intelligence (AI), have the potential to automate these processes and support more timely and accurate diagnoses. OBJECTIVES: To review current AI-based image analysis as applied to clinical microbiology; and to discuss future trends in the field. SOURCES: Material sourced for this review included peer-reviewed literature annotated in the PubMed or Google Scholar databases and preprint articles from bioRxiv. Articles describing use of AI for analysis of images used in infectious disease diagnostics were reviewed. CONTENT: We describe application of machine learning towards analysis of different types of microbiologic image data. Specifically, we outline progress in smear and plate interpretation as well as the potential for AI diagnostic applications in the clinical microbiology laboratory. IMPLICATIONS: Combined with automation, we predict that AI algorithms will be used in the future to prescreen and preclassify image data, thereby increasing productivity and enabling more accurate diagnoses through collaboration between the AI and the microbiologist. Once developed, image-based AI analysis is inexpensive and amenable to local and remote diagnostic use.

Genome-Wide Association Study of Spot Form of Net Blotch Resistance in the Upper Midwest Barley Breeding Programs.

Pyrenophora teres f. maculata, the causal agent of spot form of net blotch (SFNB), is an emerging pathogen of barley in the United States and Australia. Compared with net form of net blotch (NFNB), less is known in the U.S. Upper Midwest barley breeding programs about host resistance and quantitative trait loci (QTL) associated with SFNB in breeding lines. The main objective of this study was to identify QTL associated with SFNB resistance in the Upper Midwest two-rowed and six-rowed barley breeding programs using a genome-wide association study approach. A total of 376 breeding lines of barley were evaluated for SFNB resistance at the seedling stage in the greenhouse in Fargo in 2009. The lines were genotyped with 3,072 single nucleotide polymorphism (SNP) markers. Phenotypic evaluation showed a wide range of variability among populations from the four breeding programs and the two barley-row types. The two-rowed barley lines were more susceptible to SFNB than the six-rowed lines. Continuous distributions of SFNB severity indicate the quantitative nature of SFNB resistance. The mixed linear model (MLM) analysis, which included both population structure and kinship matrices, was used to identify significant SNP-SFNB associations. Principal component analysis was used to control false marker-trait association. The linkage disequilibrium (LD) estimates varied among chromosomes (10 to 20 cM). The MLM analysis identified 10 potential QTL in barley: SFNB-2H-8-10, SFNB-2H-38.03, SFNB-3H-58.64, SFNB-3H-78.53, SFNB-3H-91.88, SFNB-3H-117.1, SFNB-5H-155.3, SFNB-6H-5.4, SFNB-6H-33.74, and SFNB-7H-34.82. Among them, four QTL (SFNB-2H-8-10, SFNB-2H-38.03 SFNB-3H-78.53, and SFNB-3H-117.1) have not previously been published. Identification of SFNB resistant lines and QTL associated with SFNB resistance in this study will be useful in the development of barley genotypes with better SFNB resistance.

The conserved carboxyl domain of MorC, an inner membrane protein of Aggregatibacter actinomycetemcomitans, is essential for membrane function.

Morphogenesis protein C (MorC) of Aggregatibacter actinomycetemcomitans is important for maintaining the membrane morphology and integrity of the cell envelope of this oral pathogen. The MorC sequence and operon organization were found to be conserved in Gammaproteobacteria, based on a bioinformatic analysis of 435 sequences from representative organisms. Functional conservation of MorC was investigated using an A. actinomycetemcomitans morC mutant as a model system to express MorC homologs from four phylogenetically diverse representatives of the Gammaproteobacteria: Haemophilus influenzae, Escherichia coli, Pseudomonas aeruginosa, and Moraxella catarrhalis. The A. actinomycetemcomitans strains expressing the homologous proteins were assessed for sensitivity to bile salts, leukotoxin secretion, autoaggregation and membrane morphology. MorC from the most closely related organism (H. influenzae) was functionally identical to MorC from A. actinomycetemcomitans. However, the genes from more distantly related organisms restored some but not all A. actinomycetemcomitans mutant phenotypes. In addition, deletion mutagenesis indicated that the most conserved portion of the protein, the C-terminus DUF490 domain, was necessary to maintain the integrity of the membrane. Deletion of the last 10 amino acids of this domain of the A. actinomycetemcomitans MorC protein was sufficient to disrupt membrane stability and leukotoxin secretion. The data suggest that the MorC sequence is functionally conserved across Gammaproteobacteria and the C-terminus of the protein is essential for maintaining membrane physiology.

Assessing Genomic Selection Prediction Accuracy in a Dynamic Barley Breeding Population.

Prediction accuracy of genomic selection (GS) has been previously evaluated through simulation and cross-validation; however, validation based on progeny performance in a plant breeding program has not been investigated thoroughly. We evaluated several prediction models in a dynamic barley breeding population comprised of 647 six-row lines using four traits differing in genetic architecture and 1536 single nucleotide polymorphism (SNP) markers. The breeding lines were divided into six sets designated as one parent set and five consecutive progeny sets comprised of representative samples of breeding lines over a 5-yr period. We used these data sets to investigate the effect of model and training population composition on prediction accuracy over time. We found little difference in prediction accuracy among the models confirming prior studies that found the simplest model, random regression best linear unbiased prediction (RR-BLUP), to be accurate across a range of situations. In general, we found that using the parent set was sufficient to predict progeny sets with little to no gain in accuracy from generating larger training populations by combining the parent set with subsequent progeny sets. The prediction accuracy ranged from 0.03 to 0.99 across the four traits and five progeny sets. We explored characteristics of the training and validation populations (marker allele frequency, population structure, and linkage disequilibrium, LD) as well as characteristics of the trait (genetic architecture and heritability, H2 ). Fixation of markers associated with a trait over time was most clearly associated with reduced prediction accuracy for the mycotoxin trait DON. Higher trait H2 in the training population and simpler trait architecture were associated with greater prediction accuracy.

The cell envelope proteome of Aggregatibacter actinomycetemcomitans.

The cell envelope of gram-negative bacteria serves a critical role in maintenance of cellular homeostasis, resistance to external stress, and host-pathogen interactions. Envelope protein composition is influenced by the physiological and environmental demands placed on the bacterium. In this study, we report a comprehensive compilation of cell envelope proteins from the periodontal and systemic pathogen Aggregatibacter actinomycetemcomitans VT1169, an afimbriated serotype b strain. The urea-extracted membrane proteins were identified by mass spectrometry-based shotgun proteomics. The membrane proteome, isolated from actively growing bacteria under normal laboratory conditions, included 648 proteins representing 27% of the predicted open reading frames in the genome. Bioinformatic analyses were used to annotate and predict the cellular location and function of the proteins. Surface adhesins, porins, lipoproteins, numerous influx and efflux pumps, multiple sugar, amino acid and iron transporters, and components of the type I, II and V secretion systems were identified. Periplasmic space and cytoplasmic proteins with chaperone function were also identified. A total of 107 proteins with unknown function were associated with the cell envelope. Orthologs of a subset of these uncharacterized proteins are present in other bacterial genomes, whereas others are found exclusively in A. actinomycetemcomitans. This knowledge will contribute to elucidating the role of cell envelope proteins in bacterial growth and survival in the oral cavity.

Structural and functional characterization of a winter malting barley.

The development of winter malting barley (Hordeum vulgare L.) varieties is emerging as a worldwide priority due to the numerous advantages of these varieties over spring types. However, the complexity of both malting quality and winter hardiness phenotypes makes simultaneous improvement a challenge. To obtain an understanding of the relationship between loci controlling winter hardiness and malt quality and to assess the potential for breeding winter malting barley varieties, we structurally and functionally characterized the six-row accession "88Ab536", a cold-tolerant line with superior malting quality characteristics that derives from the cross of NE76129/Morex//Morex. We used 4,596 SNPs to construct the haplotype structure of 88Ab536 on which malting quality and winter hardiness loci reported in the literature were aligned. The genomic regions determining malting quality and winter hardiness traits have been defined in this founder germplasm, which will assist breeders in targeting regions for marker-assisted selection. The Barley1 GeneChip array was used to functionally characterize 88Ab536 during malting. Its gene expression profile was similar to that of the archetypical malting variety Morex, which is consistent with their similar malting quality characteristics. The characterization of 88Ab536 has increased our understanding of the genetic relationships of malting quality and winter hardiness, and will provide a genetic foundation for further development of more cold-tolerant varieties that have malt quality characteristics that meet or exceed current benchmarks.

Mapping net form net blotch and septoria speckled leaf blotch resistance Loci in barley.

Septoria speckled leaf blotch (SSLB), caused by Septoria passerinii Sacc., and net form net blotch (NB), caused by Pyrenophora teres f. teres Drechsler, are fungal diseases that decrease the yields of barley in the Upper Midwest. An effective way to manage these diseases is to plant resistant cultivars. To characterize the genetics of resistance to both pathogens, two advanced barley breeding lines, one resistant to NB (M120) and another resistant to SSLB (Sep2-72), were crossed, creating a population of 115 recombinant inbred lines. The two parents and the population were evaluated in three greenhouse seedling assays for each pathogen and for simple-sequence repeat and diversity arrays technology markers. Composite interval mapping revealed two major quantitative trait loci (QTL) associated with NB on chromosome 6H, located in bins 2 and 6. The QTL located in bin 6 explained 19 to 48% of the phenotypic variation and the QTL located in bin 2 explained 25 to 44% of the phenotypic variation. A new locus for resistance to SSLB, Rsp4, was identified on chromosome 6H, located in bins 3 to 4. Mapping these genes in elite breeding germplasm will accelerate the development and utilization of marker-assisted selection to enhance resistance to these diseases.

Genetic architecture of quantitative trait loci associated with morphological and agronomic trait differences in a wild by cultivated barley cross.

Hordeum vulgare subsp. spontaneum is the progenitor of cultivated barley (Hordeum vulgare L.). Domestication combined with plant breeding has led to the morphological and agronomic characteristics of modern barley cultivars. The objective of this study was to map the genetic factors that morphologically and agronomically differentiate wild barley from modern barley cultivars. To address this objective, we identified quantitative trait loci (QTLs) associated with plant height, flag leaf width, spike length, spike width, glume length in relation to seed length, awn length, fragility of ear rachis, endosperm width and groove depth, heading date, flag leaf length, number of tillers per plant, and kernel color in a Harrington/OUH602 advanced backcross (BC2F8) population. This population was genotyped with 113 simple sequence repeat markers. Thirty QTLs were identified, of which 16 were newly identified in this study. One to 4 QTLs were identified for each of the traits except glume length, for which no QTL was detected. The portion of phenotypic variation accounted for by individual QTLs ranged from about 9% to 54%. For traits with more than one QTL, the phenotypic variation explained ranged from 25% to 71%. Taken together, our results reveal the genetic architecture of morphological and agronomic traits that differentiate wild from cultivated barley.

Analysis of the chromosome 2(2H) region of barley associated with the correlated traits Fusarium head blight resistance and heading date.

Fusarium head blight (FHB) is a major disease of barley (Hordeum vulgare L.) that results in reduced grain yield and quality through the accumulation of the mycotoxin deoxynivalenol (DON). Coincident QTL for FHB severity, DON concentration, and heading date (HD) map to a region of chromosome 2(2H) designated Qrgz-2H-8. It is unclear whether disease resistance at this locus is due to a pleiotropic effect of late HD by delaying the host exposure to the pathogen or a tightly linked resistance gene. The objectives of this study were to develop a set of near isogenic lines (NILs) for the Qrgz-2H-8 region and to genetically dissect the QTL region containing the coincident traits. Two NIL populations were developed consisting of F(2)- and F(4)-derived recombinants from a cross between a BC(5) line carrying the donor parent (Chevron) alleles in the Qrgz-2H-8 region and the recurrent parent M69. Analysis of field and marker data from these NILs revealed that the Chevron alleles conditioning FHB resistance, late HD, and low DON concentration were successfully introgressed into the BC(5) parent line and were segregating among NILs. QTL analysis of the F(4)-derived population showed that the HD QTL is adjacent to the FHB QTL. Furthermore, a single NIL was identified that was similar to the resistant BC(5) parent for FHB severity and the early flowering parent M69 for HD. These results indicate that the relationship between FHB and HD at the Qrgz-2H-8 region is likely due to tight linkage rather than pleiotropy.

Exploiting selective genotyping to study genetic diversity of resistance to Fusarium head blight in barley.

Numerous barley cultivars from around the world have been identified as potential sources of Fusarium head blight (FHB) resistance genes. All of these cultivars exhibit partial resistance, and several mapping studies have shown that resistance to FHB is controlled by multiple genes. Successful development of barley cultivars with high levels of FHB resistance will require combining genes from multiple sources. We characterized five potential new sources of FHB resistance ('AC Oxbow', 'Atahualpa', 'HOR211', 'PFC88209', and 'Zhedar#1') to determine if they contain new FHB resistance genes. Cluster analysis, using a set of 80 SSR markers distributed throughout the genome, showed that most of the new sources of resistance were not similar to three cultivars that have been used in previous FHB mapping studies ('Chevron', 'Frederickson', and 'Gobernadora'), with 'Atahualpa' and 'HOR211' being the most dissimilar. By selective genotyping, we determined whether markers linked to six known FHB resistance quantitative trait loci (QTLs), discovered in other genotypes, explained variation for resistance in advanced breeding populations created from the new sources of resistance. Markers linked to four of the six known QTLs were associated with FHB severity in at least one of the populations. However, none of the six QTL regions were associated with variation for FHB severity in populations derived from crosses that utilized sources of resistance HOR211 or PFC88209. Selective genotyping is an efficient method for breeders to utilize current QTL information about disease resistance to search for new resistance genes.

Host genetic effect on deoxynivalenol accumulation in fusarium head blight of barley.

ABSTRACT One of the major concerns with Fusarium head blight (FHB) of barley is the potential health risks to livestock and humans through the accumulation of the mycotoxin deoxynivalenol (DON) in infected grain. To define the role of the host in DON accumulation during the early stages of disease development, we conducted a series of greenhouse experiments. We inoculated single spikelets of greenhouse-grown plants with Fusarium graminearum, moved the plants to a dew chamber, and harvested the inoculated spikelets after 72 h for DON analysis. We conducted a quantitative trait locus (QTL) analysis using a genetic mapping population, constructed with the parents Stander and Frederickson, that segregated for DON accumulation after single-spikelet inoculation in two experiments. A single QTL on chromosome 3 explained 18 and 35% of the phenotypic variation in the two experiments. To validate this QTL for DON accumulation, we used a DNA marker to select near-isogenic lines from a family from the mapping population that was segregating at this QTL. Disease symptom development was similar between the nearisogenic lines; however, the mean DON concentration of the lines homozygous for the allele from the high DON parent was 2.5-fold more than the lines homozygous for the alternate allele. A time course experiment showed that this effect on toxin accumulation was observed at 10 days post inoculation. The near-isogenic lines developed in this study should prove useful for further exploration of the role of DON in FHB.

Upper gastrointestinal hemorrhage and transcatheter embolotherapy: clinical and technical factors impacting success and survival.

PURPOSE: To identify clinical and technical factors influencing the outcome of transcatheter embolotherapy for nonvariceal upper gastrointestinal (GI) hemorrhage and to quantify the impact of successful intervention on patient survival. MATERIALS AND METHODS: A retrospective review was performed of all patients (n = 163) who underwent arterial embolization for acute upper GI hemorrhage at a university hospital over an 11.5-year period. Clinical success was defined as target area devascularization that resulted in the clinical cessation of bleeding and stabilization of hemoglobin level. The clinical condition of each patient at intervention was defined by history, laboratory examination, and two composite indicator variables. With use of logistic regression, the dependent variable, clinical success, was modeled on two categories of clinical and technical variables. A final model regressed patient survival on clinical success and other clinical variables. RESULTS: None of the procedural variables analyzed had a significant influence on clinical success. Several clinical variables did impact clinical success, including multiorgan system failure (OR, 0.36; P =.030), coagulopathy (OR, 0.36; P =.026), and bleeding subsequent to trauma (OR, 7.1; P =.040) or invasive procedures (OR, 6.5; P =.009). Regardless of their clinical condition at intervention, patients who underwent clinically successful embolization were 13.3 times more likely to survive than those who had an unsuccessful procedure (CI, 4.54-39.2; P =.000). Nevertheless, patients with multiorgan system failure were 17.5 times more likely to die, independent of the outcome of the procedure (CI, 0.014-0.229; P =.000). CONCLUSION: Arresting nonvariceal upper GI hemorrhage with transcatheter embolotherapy has a large positive effect on patient survival, independent of clinical condition or demonstrable extravasation at intervention. Aggressive treatment with transcatheter embolotherapy is advisable in patients with acute nonvariceal upper GI hemorrhage.

Influence of tomato genotype on growth of inoculated and indigenous bacteria in the spermosphere.

We previously demonstrated a genetic basis in tomato for support of the growth of a biological control agent, Bacillus cereus UW85, in the spermosphere after seed inoculation (K. P. Smith, J. Handelsman, and R. M. Goodman, Proc. Natl. Acad. Sci. USA 96:4786-4790, 1999). Here we report results of studies examining the host effect on the support of growth of Bacillus and Pseudomonas strains, both inoculated on seeds and recruited from soil, using selected inbred tomato lines from the recombinant inbred line (RIL) population used in our previous study. Two tomato lines, one previously found to support high and the other low growth of B. cereus UW85 in the spermosphere, had similar effects on growth of each of a diverse, worldwide collection of 24 B. cereus strains that were inoculated on seeds and planted in sterilized vermiculite. In contrast, among RILs that differed for support of B. cereus UW85 growth in the spermosphere, we found no difference for support of growth of the biocontrol strains Pseudomonas fluorescens 2-79 or Pseudomonas aureofaciens AB254. Thus, while the host effect on growth extended to all strains of B. cereus examined, it was not exerted on other bacterial species tested. When seeds were inoculated with a marked mutant of B. cereus UW85 and planted in soil, RIL-dependent high and low support of bacterial growth was observed that was similar to results from experiments conducted in sterilized vermiculite. When uninoculated seeds from two of these RILs were planted in soil, changes in population levels of indigenous Bacillus and fluorescent Pseudomonas bacteria differed, as measured over time by culturing and direct microscopy, from growth patterns observed in the inoculation experiments. Neither RIL supported detectable levels of growth of indigenous Bacillus soil bacteria, while the line that supported growth of inoculated B. cereus UW85 supported higher growth of indigenous fluorescent pseudomonads and total bacteria. The vermiculite system used in these experiments was predictive for growth of B. cereus UW85 inoculated on seeds and grown in soil, but the patterns of growth of inoculated strains-both Bacillus and Pseudomonas spp.-did not reflect host genotype effects on indigenous microflora recruited from soil to the spermosphere.

Interactions of U2 gene loci and their nuclear transcripts with Cajal (coiled) bodies: evidence for PreU2 within Cajal bodies.

The Cajal (coiled) body (CB) is a structure enriched in proteins involved in mRNA, rRNA, and snRNA metabolism. CBs have been shown to interact with specific histone and snRNA gene loci. To examine the potential role of CBs in U2 snRNA metabolism, we used a variety of genomic and oligonucleotide probes to visualize in situ newly synthesized U2 snRNA relative to U2 loci and CBs. Results demonstrate that long spacer sequences between U2 coding repeats are transcribed, supporting other recent evidence that U2 transcription proceeds past the 3' box. The presence of bright foci of this U2 locus RNA differed between alleles within the same nucleus; however, this did not correlate with the loci's association with a CB. Experiments with specific oligonucleotide probes revealed signal for preU2 RNA within CBs. PreU2 was also detected in the locus-associated RNA foci, whereas sequences 3' of preU2 were found only in these foci, not in CBs. This suggests that a longer primary transcript is processed before entry into CBs. Although this work shows that direct contact of a U2 locus with a CB is not simply correlated with RNA at that locus, it provides the first evidence of new preU2 transcripts within CBs. We also show that, in contrast to CBs, SMN gems do not associate with U2 gene loci and do not contain preU2. Because other evidence indicates that preU2 is processed in the cytoplasm before assembly into snRNPs, results point to an involvement of CBs in modification or transport of preU2 RNA.

Genetic basis in plants for interactions with disease-suppressive bacteria.

Plant health depends, in part, on associations with disease-suppressive microflora, but little is known about the role of plant genes in establishing such associations. Identifying such genes will contribute to understanding the basis for plant health in natural communities and to new strategies to reduce dependence on pesticides in agriculture. To assess the role of the plant host in disease suppression, we used a genetic mapping population of tomato to evaluate the efficacy of the biocontrol agent Bacillus cereus against the seed pathogen Pythium torulosum. We detected significant phenotypic variation among recombinant inbred lines that comprise the mapping population for resistance to P. torulosum, disease suppression by B. cereus, and growth of B. cereus on the seed. Genetic analysis revealed that three quantitative trait loci (QTL) associated with disease suppression by B. cereus explained 38% of the phenotypic variation among the recombinant inbred lines. In two cases, QTL for disease suppression by B. cereus map to the same locations as QTL for other traits, suggesting that the host effect on biocontrol is mediated by different mechanisms. The discovery of a genetic basis in the host for interactions with a biocontrol agent suggests new opportunities to exploit natural genetic variation in host species to enhance our understanding of beneficial plant-microbe interactions and develop ecologically sound strategies for disease control in agriculture.

Processing of endogenous pre-mRNAs in association with SC-35 domains is gene specific.

Analysis of six endogenous pre-mRNAs demonstrates that localization at the periphery or within splicing factor-rich (SC-35) domains is not restricted to a few unusually abundant pre-mRNAs, but is apparently a more common paradigm of many protein-coding genes. Different genes are preferentially transcribed and their RNAs processed in different compartments relative to SC-35 domains. These differences do not simply correlate with the complexity, nuclear abundance, or position within overall nuclear space. The distribution of spliceosome assembly factor SC-35 did not simply mirror the distribution of individual pre-mRNAs, but rather suggested that individual domains contain both specific pre-mRNA(s) as well as excess splicing factors. This is consistent with a multifunctional compartment, to which some gene loci and their RNAs have access and others do not. Despite similar molar abundance in muscle fiber nuclei, nascent transcript "trees" of highly complex dystrophin RNA are cotranscriptionally spliced outside of SC-35 domains, whereas posttranscriptional "tracks" of more mature myosin heavy chain transcripts overlap domains. Further analyses supported that endogenous pre-mRNAs exhibit distinct structural organization that may reflect not only the expression and complexity of the gene, but also constraints of its chromosomal context and kinetics of its RNA metabolism.

Quantitative trait loci associated with resistance to Fusarium head blight and kernel discoloration in barley.

Resistance to Fusarium head blight (FHB), deoxynivalenol (DON) accumulation, and kernel discoloration (KD) in barley are difficult traits to introgress into elite varieties because current screening methods are laborious and disease levels are strongly influenced by environment. To improve breeding strategies directed toward enhancing these traits, we identified genomic regions containing quantitative trait loci (QTLs) associated with resistance to FHB, DON accumulation, and KD in a breeding population of F(4:7) lines using restriction fragment length polymorphic (RFLP) markers. We evaluated 101 F(4:7) lines, derived from a cross between the cultivar Chevron and an elite breeding line, M69, for each of the traits in three or four environments. We used 94 previously mapped RFLP markers to create a linkage map. Using composite interval mapping, we identified 10, 11, and 4 QTLs associated with resistance to FHB, DON accumulation, and KD, respectively. Markers flanking these QTLs should be useful for introgressing resistance to FHB, DON accumulation, and KD into elite barley cultivars.

Cloning, characterization, and chromosomal localization of human superillin (SVIL).

Supervillin is a 205-kDa F-actin binding protein originally isolated from bovine neutrophils. This protein is tightly associated with both actin filaments and plasma membranes, suggesting that it forms a high-affinity link between the actin cytoskeleton and the membrane. Human supervillin cDNAs cloned from normal human kidney and from the cervical carcinoma HeLa S3 predict a bipartite structure with three potential nuclear localization signals in the NH2-terminus and three potential actin-binding sequences in the COOH-terminus. In fact, throughout its length, the COOH-terminal half of supervillin is similar to segments 2-6 plus the COOH-terminal "headpiece" of villin, an actin-binding protein in intestinal microvilli. A comparison of the bovine and human sequences indicates that supervillin is highly conserved at the amino acid level, with 79.2% identity of the NH2-terminus and conservation of three of the four nuclear localization signals found in bovine supervillin. The COOH-terminus is even more conserved, with 95.1% amino acid identity overall and 100% conservation of the villin-like headpiece. Supervillin mRNAs are expressed in all human tissue tested, bu are most abundant in muscle, bone marrow, thyroid gland, and salivary gland; comparatively little message is found in brain. Human supervillin mRNA is approximately 7.5 kb; this message is especially abundant in HeLa S3 cervical carcinoma, SW480 adenocarcinoma, and A549 lung carcinoma cell lines. The human supervillin gene (SVIL) is localized to a single chromosomal locus at 10p11.2, a region that is deleted in some prostate tumors.