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J H Hill

Publications and source records attributed to J H Hill.

At least 19 recordsLinked to original sources
PubMed2005

Strain-specific P3 of Soybean mosaic virus elicits Rsv1-mediated extreme resistance, but absence of P3 elicitor function alone is insufficient for virulence on Rsv1-genotype soybean.

When challenged by mechanical inoculation, the Rsv1 gene of soybean invokes extreme resistance (ER) against Soybean mosaic virus (SMV) strain N, but not SMV-G7 and its experimentally evolved variant, SMV-G7d. SMV-G7 provokes a lethal systemic hypersensitive response (LSHR), whereas SMV-G7d induces systemic mosaic. Thus, for Rsv1-genotype soybean, SMV-G7 and SMV-G7d are both virulent virus strains. The elicitor function of SMV-G7 provoking Rsv1-mediated LSHR was recently mapped to P3, and the influence of amino acids 823, 953, and 1112 of the precursor polypeptide of SMV-G7d on evasion of Rsv1-mediated recognition provoking LSHR was demonstrated. We have now extended this study to SMV-N. Initially, amino acids corresponding to those of SMV-G7d at these positions were substituted, individually or in combinations. All the mutants remained replication competent on rsv1-genotype soybean; however, none lost the elicitor function provoking Rsv1-mediated ER. Subsequently, P3 of SMV-N was precisely replaced with P3 of SMV-G7 or SMV-G7d and vice versa. All the chimeras were replication competent on rsv1-genotype soybean, but surprisingly SMV-N/G7P3 and SMV-N/G7dP3 failed to gain virulence on Rsv1-genotype soybeans. However, SMV-G7/NP3 and SMV-G7d/NP3 lost virulence, and this loss of virulence function was mapped to the N-terminus domain of SMV-N P3. The data indicate that SMV strain-specific P3 provokes Rsv1-mediated ER; however, virulence on Rsv1-genotype soybean is not solely a consequence of the absence of the P3 elicitor functions provoking Rsv1-mediated ER and LSHR.

Amino Acid Sequence↗
PubMed2005

Loss and gain of elicitor function of soybean mosaic virus G7 provoking Rsv1-mediated lethal systemic hypersensitive response maps to P3.

Rsv1, a single dominant resistance gene in soybean PI 96983 (Rsv1), confers extreme resistance against all known American strains of Soybean mosaic virus (SMV), except G7 and G7d. SMV-G7 provokes a lethal systemic hypersensitive response (LSHR), whereas SMV-G7d, an experimentally evolved variant of SMV-G7, induces systemic mosaic. To identify the elicitor of Rsv1-mediated LSHR, chimeras were constructed by exchanging fragments between the molecularly cloned SMV-G7 (pSMV-G7) and SMV-G7d (pSMV-G7d), and their elicitor functions were assessed on PI 96983 (Rsv1). pSMV-G7-derived chimeras containing only P3 of SMV-G7d lost the elicitor function, while the reciprocal chimera of pSMV-G7d gained the function. The P3 regions of the two viruses differ by six nucleotides, of which two are translationally silent. The four amino acid differences are located at positions 823, 915, 953, and 1112 of the precursor polypeptide. Analyses of the site-directed point mutants of both the viruses revealed that nucleotide substitutions leading to translationally silent mutations as well as reciprocal amino acid substitution at position 915 did not influence the loss or gain of the elicitor function. pSMV-G7-derived mutants with amino acid substitutions at any of the other three positions lost the ability to provoke LSHR but induced SHR instead. Two concomitant amino acid substitutions at positions 823 (V to M) and 953 (K to E) abolished pSMV-G7 elicitor function, provoking Rsv1-mediated SHR. Conversely, pSMV-G7d gained the elicitor function of Rsv1-mediated LSHR by a single amino acid substitution at position 823 (M to V), and mutants with amino acid substitutions at position 953 or 1112 induced SHR instead of mosaic. Taken together, the data suggest that strain-specific P3 of SMV is the elicitor of Rsv1-mediated LSHR.

Base Sequence↗
PubMed2003

Evolution of Soybean mosaic virus-G7 molecularly cloned genome in Rsv1-genotype soybean results in emergence of a mutant capable of evading Rsv1-mediated recognition.

Plant resistance (R) genes direct recognition of pathogens harboring matching avirluent signals leading to activation of defense responses. It has long been hypothesized that under selection pressure the infidelity of RNA virus replication together with large population size and short generation times results in emergence of mutants capable of evading R-mediated recognition. In this study, the Rsv1/Soybean mosaic virus (SMV) pathosystem was used to investigate this hypothesis. In soybean line PI 96983 (Rsv1), the progeny of molecularly cloned SMV strain G7 (pSMV-G7) provokes a lethal systemic hypersensitive response (LSHR) with up regulation of a defense-associated gene transcript (PR-1). Serial passages of a large population of the progeny in PI 96983 resulted in emergence of a mutant population (vSMV-G7d), incapable of provoking either Rsv1-mediated LSHR or PR-1 protein gene transcript up regulation. An infectious clone of the mutant (pSMV-G7d) was synthesized whose sequences were very similar but not identical to the vSMV-G7d population; however, it displayed a similar phenotype. The genome of pSMV-G7d differs from parental pSMV-G7 by 17 substitutions, of which 10 are translationally silent. The seven amino acid substitutions in deduced sequences of pSMV-G7d differ from that of pSMV-G7 by one each in P1 proteinase, helper component-proteinase, and coat protein, respectively, and by four in P3. To the best of our knowledge, this is the first demonstration in which experimental evolution of a molecularly cloned plant RNA virus resulted in emergence of a mutant capable of evading an R-mediated recognition.

Cloning, Molecular↗
PubMed2001

Rsv1-mediated resistance against soybean mosaic virus-N is hypersensitive response-independent at inoculation site, but has the potential to initiate a hypersensitive response-like mechanism.

Rsv1, a single dominant gene in soybean PI 96983, confers resistance to most strains of Soybean mosaic virus (SMV), including strain G2. The phenotypic response includes the lack of symptoms and virus recovery from mechanically inoculated leaves. To study the resistance mechanism, SMV-N (an isolate of strain G2) was introduced into PI 96983 by grafting. Hypersensitive response (HR)-like lesions occurred on the stems, petioles, and leaf veins, and virus was recovered from these lesions. The response demonstrated the cytological and histological characteristics of HR as well as elevated transcription of a soybean salicylic acid-inducible, pathogenesis-related (PR-1) protein gene. Mechanical inoculation of PI 96983 primary leaves with a high level of SMV-N virions caused no symptoms or up regulation of the PR-1 protein gene transcript. Furthermore, inoculation with infectious viral RNA did not alter the resistance phenotype. The data suggest that interaction of SMV-N with Rsv1 has the potential to induce an HR-like defense reaction. Rsv1-mediated resistance in the inoculated leaf, however, is HR-independent and operates after virion disassembly.

Genes, Dominant↗
PubMed1999

Intragenic suppression of an active site mutation in the human apurinic/apyrimidinic endonuclease.

The apurinic/apyrimidinic endonucleases (APE) contain several highly conserved sequence motifs. The glutamic acid residue in a consensus motif, LQE96TK98 in human APE (hAPE-1), is crucial because of its role in coordinating Mg2+, an essential cofactor. Random mutagenesis of the inactive E96A mutant cDNA, followed by phenotypic screening in Escherichia coli, led to isolation of an intragenic suppressor with a second site mutation, K98R. Although the Km of the suppressor mutant was about sixfold higher than that of the wild-type enzyme, their kcat values were similar for AP endonuclease activity. These results suggest that the E96A mutation affects only the DNA-binding step, but not the catalytic step of the enzyme. The 3' DNA phosphoesterase activities of the wild-type and the suppressor mutant were also comparable. No global change of the protein conformation is induced by the single or double mutations, but a local perturbation in the structural environment of tryptophan residues may be induced by the K98R mutation. The wild-type and suppressor mutant proteins have similar Mg2+ requirement for activity. These results suggest a minor perturbation in conformation of the suppressor mutant enabling an unidentified Asp or Glu residue to substitute for Glu96 in positioning Mg2+ during catalysis. The possibility that Asp70 is such a residue, based on its observed proximity to the metal-binding site in the wild-type protein, was excluded by site-specific mutation studies. It thus appears that another acidic residue coordinates with Mg2+ in the mutant protein. These results suggest a rather flexible conformation of the region surrounding the metal binding site in hAPE-1 which is not obvious from the X-ray crystallographic structure.

Carbon-Oxygen Lyases↗
PubMed1998

Characterization of a monoclonal antibody recognizing a DAG-containing epitope conserved in aphid transmissible potyviruses: evidence that the DAG motif is in a defined conformation.

Two viral proteins, the helper component-protease and the coat protein, are required for the non-persistent aphid transmission of potyviruses. In the potyvirus coat protein, the tripeptide aspartate-alanine-glycine (DAG) has often been shown to be involved. A monoclonal antibody, raised against a synthetic decapeptide containing the DAG tripeptide, reacted with the peptide as well as with isolates of soybean mosaic, tobacco etch and tobacco vein mottling potyviruses. Experiments indicate that the monoclonal antibody recognizes a conformational rather than a sequential epitope. The data support the suggestion that the DAG region plays a structural role to determine a coat protein-helper component-protease conformation that influences aphid transmission.

Amino Acid Sequence↗
PubMed1997

CO2 enrichment influences yields of 'Florunner,' 'Georgia Red' and 'New Mexico' peanut cultivars.

Three peanut cultivars, 'Florunner,' 'Georgia Red,' and 'New Mexico,' were grown in reach-in chambers to determine response to CO2 enrichment. CO2 treatments were ambient (400 micromol mol-1) and 700 micromol mol-1. Growth chamber conditions included 700 micromol m-2 s-1 photosynthetic photon flux (PPF), 28/22C, 7O% RH, and 12/12 h photoperiod. Growth media consisted of a 1:1 mixture (v/v) of vermiculite and sterilized sand. Six 10 L pots of each cultivar were fertilized three times per week with 250 mL of nutrient solution containing additional Ca (10 mM) and NO3 (25 mM) and watered well. Beginning 21 days after planting (DAP) and every three weeks thereafter up to 84 days, the second leaf from the growing axis (main stem) was detached to determine CO2 effect on leaf area, specific leaf area (SLA) and dry weight. Plants were harvested 97 DAP, at which time total leaf area, leaf number, plant and root weights and pod production data were taken. Numbers of pods per plant, pod fresh and dry weights, fibrous root and plant dry weights were higher for all cultivars grown at 700 micromol mol-1 than at ambient CO2. Also, leaf area for all cultivars was larger with CO2 enrichment than at ambient. SLA tended to decline with time regardless of CO2 treatment. Percentage of total sound mature kernels (%TSMK) was similar for both treatments. Plants grown at 700 micromol mol-1 CO2 had slightly more immature pods and seeds at final harvest.

Arachis↗
PubMed1993

Molecular basis for virus disease resistance in plants.

Classical studies of virus disease resistance in plants have provided the basis for recent molecular studies of resistance. Three common approaches to the study of resistance have been used. In one approach, nucleotide and/or amino acid sequences of virus strains that overcome disease resistance genes in the host are compared with sequences of strains that do not induce disease in these hosts. In the second approach, resistance/susceptibility of protoplasts is compared with the response of intact plants from which they are derived, to develop hypotheses regarding whether resistance acts at the level of the individual cell or by inhibiting cell-to-cell movement. In the third approach, the mechanism of virus cell-to-cell movement has been studied to clarify one of the basic steps in pathogenesis and to determine the mechanism of disease resistance for certain virus-host interactions.

Immunity, Innate↗
PubMed1993

Antigenic signature analysis reflects differences among plant virus isolates.

Antigenic differences among cowpea severe mosaic virus (CPSMV) isolates were clearly reflected in signature analysis employing a panel of seven well-characterized, monoclonal antibodies. Separate binding curves were generated by reacting serial dilutions of extracts from infected plant tissue containing each antigen simultaneously with each antibody in the panel. An iterative procedure was used to align unknown CPSMV antigen concentrations from different antigen preparations to allow comparison of binding profiles from different assays. Signature analysis was shown to be highly useful for the elucidation of subtle antigenic differences among viral agents because it requires neither purified virus nor knowledge of virus concentration in sap from infected plants.

Antibodies, Monoclonal↗
PubMed1992

Complete nucleotide sequences of two soybean mosaic virus strains differentiated by response of soybean containing the Rsv resistance gene.

The complete nucleotide sequence of the genomic RNAs of strains G2 and G7 of soybean mosaic virus were determined. In both cases, the genome is 9588 nucleotides long, excluding the 3'-terminal poly(A) sequence. A large open reading frame (nucleotides 132 to 9329) encodes a polyprotein of 3066 amino acids with a predicted M(r) of either 349542 (strain G2) or 349741 (strain G7). Based on comparison with the proposed locations of cleavage sites of other potyvirus polyproteins, nine mature proteins are predicted. The mature proteins of the two strains share 94 to 100% amino acid identity, with the greatest variability occurring in the 35K and 42K proteins. Differences in local net charge in portions of these proteins as well as differences in amino acid sequence throughout the genome are discussed in relation to resistance and susceptibility of host plants to strains G2 and G7. Comparison with other potyviruses may be useful for taxonomic clarification of viruses and strains.

Amino Acid Sequence↗
PubMed1991

Effect of intracanalicular collagen implants on the absorption of topically applied sodium fluorescein.

Absorbable intracanalicular collagen implants were placed in both canaliculi of one eye of nine human volunteers. The other eye served as a control. Twenty-four hours later 2% sodium fluorescein was placed into both conjunctival sacs. Serial corneal fluorescein concentrations were measured with a scanning ocular fluorophotometer from 2 to 90 min after fluorescein administration. The mean corneal fluorescein concentration averaged over all time points was greater in 7 of 9 eyes with collagen implants compared to unimplanted controls. When the data from each subject were analyzed collectively, the mean corneal fluorescein concentration in the implanted eyes (1,218 micrograms/ml +/- SEM 83) was significantly greater (p less than 0.001) than the mean concentration in the control eyes (823 micrograms/ml +/- SEM 83). The use of absorbable intracanalicular collagen implants may increase the bioavailability of topically applied ocular solutions.

Absorption↗
PubMed1991

Nucleotide sequences of the coat protein genes of two aphid-transmissible strains of soybean mosaic virus.

The nucleotide sequences of the coat protein genes and 3' non-coding regions of two aphid-transmissible isolates (G2 and G7) of soybean mosaic virus (SMV) were determined. The coat protein of the G2 isolate differs from that of the aphid non-transmissible N isolate by a single amino acid at position 12 (aspartic acid in N, glycine in G2 and G7). The G7 isolate differs from G2 and N at three and four amino acid residues, respectively. The nucleotide sequence similarity of the three isolates in the coat protein coding and 3' non-coding regions ranges from 93 to 100%, respectively. In contrast, watermelon mosaic virus 2 (WMV 2) is only 77% to 79% similar to the SMV isolates, suggesting that WMV 2 is a distinct virus and not an isolate of SMV as proposed previously.

Amino Acid Sequence↗
PubMed1991

A community's response to childhood drownings. A model for accident prevention.

The prevention of drowning or any other accidental injury depends primarily on changing the behavior patterns of the families of the children at risk. Many community organizations and individual citizens must be recruited to help convince the public of the risks of young children drowning in backyard pools and to propose appropriate prevention measures. Legally mandated remedies may be necessary and may take years to show any significant effect. Because of the constantly changing population of parents of young children and the high risk of drowning for small children in Arizona, the educational programs must be repeated year after year.

Arizona↗
PubMed1990

Genetic engineering of plants for virus resistance.

Historically, control of plant virus disease has involved numerous strategies which have often been combined to provide effective durable resistance in the field. In recent years, the dramatic advances obtained in plant molecular virology have enhanced our understanding of viral genome organizations and gene functions. Moreover, genetic engineering of plants for virus resistance has recently provided promising additional strategies for control of virus disease. At present, the most promising of these has been the expression of coat-protein coding sequences in plants transformed with a coat protein gene. Other potential methods include the expression of anti-sense viral transcripts in transgenic plants, the application of artificial anti-sense mediated gene regulation to viral systems, and the expression of viral satellite RNAs, RNAs with endoribonuclease activity, antiviral antibody genes, or human interferon genes in plants.

Base Sequence↗
PubMed1988

Blood thiamine levels in normal cattle and sheep at pasture.

Analysis of whole blood samples from 174 cattle and, 174 sheep from 3 geographical regions of New Zealand over a 10 month period showed a mean (range) thiamine level of 122 nmol/l (71-237 nmol/l) for cattle and 118 nmol/l (67-227 nmol/l) for sheep. Regional and seasonal differences were noted with levels tending to rise over the summer period. A reference range of 75-185 nmol/l is proposed for both cattle and sheep to cover these variations. Levels below 50 nmol/l are considered indicative of deficiency.

Journal Article↗