Low-temperature damage and subsequent recovery of fab1 mutant Arabidopsis exposed to 2 degrees C.

The fab1 mutant of Arabidopsis thaliana, which contains increased levels of saturated fatty acids, was indistinguishable from the wild type when it was grown at 22 or 12 degrees C. During the first 7 to 10 d after transfer to 2 degrees C, the growth and photosynthetic characteristics of the fab1 plants remained indistinguishable from the wild type, with values for the potential quantum efficiency of photosystem II decreasing from 0.8 to 0.7 in plants of both lines. Whereas wild-type plants maintained quantum efficiency of photosystem II at approximately 0.7 for at least 35 d at 2 degrees C, this parameter declined rapidly in the mutant after 7 d and reached a value of less than 0.1 after 28 d at 2 degrees C. This decline in photosynthetic capacity was accompanied by reductions in chlorophyll content and the amount of chloroplast glycerolipids per gram of leaf. Electron microscopic examination of leaf samples revealed a rapid and extensive disruption of the thylakoid and chloroplast structure in the mutant, which is interpreted here as a form of selective autophagy. Despite the almost complete loss of photosynthetic function and the destruction of photosynthetic machinery, fab1 plants retained a substantial capacity for recovery following transfer to 22 degrees C. These results provide a further demonstration of the importance of chloroplast membrane unsaturation to the proper growth and development of plants at low temperature.

An octadecanoid pathway mutant (JL5) of tomato is compromised in signaling for defense against insect attack.

The activation of defense genes in tomato plants has been shown to be mediated by an octadecanoic acid-based signaling pathway in response to herbivore attack or other mechanical wounding. We report here that a tomato mutant (JL5) deficient in the activation of would-inducible defense genes is also compromised in resistance toward the lepidopteran predator Manduca sexta (tobacco hornworm). Thus we propose the name defenseless1 (def1) for the mutation in the JL5 line that mediates this altered defense response. In experiments designed to define the normal function of DEF1, we found that def1 plants are defective in defense gene signaling initiated by prosystemin overexpression in transgenic plants as well as by oligosaccharide (chitosan and polygalacturonide) and polypeptide (systemin) elicitors. Supplementation of plants through their cut stems with intermediates of the octadecanoid pathway indicates that def1 plants are affected in octadecanoid metabolism between the synthesis of hydroperoxylinolenic acid and 12-oxo-phytodienoic acid. Consistent with this defect, def1 plants are also compromised in their ability to accumulate jasmonic acid, the end product of the pathway, in response to wounding and the aforementioned elicitors. Taken together, these results show that octadecanoid metabolism plays an essential role in the transduction of upstream would signals to the activation of antiherbivore plant defenses.

A Mutant of Arabidopsis with Increased Levels of Stearic Acid.

A mutation at the fab2 locus of Arabidopsis caused increased levels of stearate in leaves. The increase in leaf stearate in fab2 varied developmentally, and the largest increase occurred in young leaves, where stearate accounted for almost 20% of total leaf fatty acids. The fatty acid composition of leaf lipids isolated from the fab2 mutant showed increased stearate in all the major glycerolipids of both the chloroplast and extrachloroplast membranes. Although the stearate content was increased, the fab2 mutant still contained abundant amounts of 18:1, 18:2, and 18:3 fatty acids. These results are consistent with the expectations for a mutation partially affecting the action of the stromal stearoyl-acyl carrier protein desaturase. Positional analysis indicated that the extra 18:0 is excluded with high specificity from the sn-2 position of both chloroplast and extrachloroplast glycerolipids. Although stearate content was increased in all the major leaf membrane lipids, the amount of increase varied considerably among the different lipids, from a high of 25% of fatty acids in phosphatidylcholine to a low of 2.9% of fatty acids in monogalactosyldiacylglycerol.

Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis.

The polyunsaturated fatty acids linoleate and alpha-linolenate are important membrane components and are the essential fatty acids of human nutrition. The major enzyme responsible for the synthesis of these compounds is the plant oleate desaturase of the endoplasmic reticulum, and its activity is controlled in Arabidopsis by the fatty acid desaturation 2 (fad2) locus. A fad2 allele was identified in a population of Arabidopsis in which mutations had been created by T-DNA insertions. Genomic DNA flanking the T-DNA was cloned by plasmid rescue and used to isolate cDNA and genomic clones of FAD2. A cDNA containing the entire FAD2 coding sequence was expressed in fad2 mutant plants and shown to complement the mutant fatty acid phenotype. The deduced amino acid sequence from the cDNA showed homology to other plant desaturases, and this confirmed that FAD2 is the structural gene for the desaturase. Gel blot analyses of FAD2 mRNA levels showed that the gene is expressed throughout the plant and suggest that transcript levels are in excess of the amount needed to account for oleate desaturation. Sequence analysis identified histidine-rich motifs that could contribute to an iron binding site in the cytoplasmic domain of the protein. Such a position would facilitate interaction between the desaturase and cytochrome b5, which is the direct source of electrons for the desaturation reaction, but would limit interaction of the active site with the fatty acyl substrate.

Isolation of signaling mutants of tomato (Lycopersicon esculentum).

As a first step towards developing a genetic system for investigating signaling processes in plants, we have developed a screen for signaling mutants deficient in a wound response. We have isolated two mutants of tomato that lack detectable production of proteinase inhibitors induced systemically in leaves by wounding. The mutants are deficient in the induction of both proteinase Inhibitor I and proteinase Inhibitor II but can be induced to respond at near wild-type levels by methyl jasmonate, a known elicitor of inhibitor production in tomato. While completely deficient in systemic production of proteinase inhibitors, both mutants produce some proteinase inhibitor in wounded leaves. This evidence suggests the existence of two signaling pathways, one local and one systemic, that regulate the induction of proteinase inhibitor synthesis in response to wounding.

Estimation of the costs of mastitis, using National Animal Health Monitoring System and milk somatic cell count data.

Data collected by the National Animal Health Monitoring System in Ohio for a 12-month period during 1986 and 1987 were used to determine the relative magnitude of costs associated with mastitis in the following categories: milk production loss, veterinary services, drugs, producer labor, and "other" factors. The cost of milk loss associated with mastitis that was reported by producers cooperating in the National Animal Health Monitoring System program was compared with estimates based on bulk tank somatic cell counts and individual cow milk somatic cell counts. Using producer-reported estimates, milk loss accounted for about one third of the total cost associated with mastitis. When estimates of milk loss were replaced by estimates based on bulk tank somatic cell counts, milk loss accounted for over 80% of the total cost of mastitis. Estimates of the cost of milk loss based on studies relating milk yield to somatic cell counts differed considerably. Consequently, it was unclear how to best estimate the relative magnitude of the milk loss component of mastitis costs.

Competition of internal and external information in an exercise setting.

During physical exercise, individuals have access to internal sensory and external environmental cues that compete for attentional focus. Two experiments examined when attention to external cues attenuates the perception of physical symptoms and fatigue. In Experiment 1, subjects' physical performance was held constant during exercise on a treadmill. Subjects hearing distracting sounds reported less fatigue and fewer symptoms than subjects hearing an amplification of their own breathing. In Experiment 2, subjects jogging equal length cross-country and lap courses evinced faster times on the former, where increased external attention was necessary. Self-reports of symptoms and fatigue, however, were comparable on the two courses. The results are interpreted in terms of attentional focus shifting from one information source to another as needed, with attention to any one source diminishing attention to others.

An attorney's approach to psychiatrists in custody cases.

The contested custody case has in common with any other case that it is a search for the truth through the medium of the courts. Many modern thinkers believe that the courts are not the proper arena for discovering the truth in all cases. Increasingly, there is agitation to remove areas of testimony from the courts, placing them perhaps in the hands of some commission appointed by government. So far, this has not happened in the area of child custody, and we still have the courtroom technique of examination and cross-examination, with a final decision by a judge on the basic subject of truth, that is, what is the best interest of the child. The attorney can learn from the doctor in this process, and I think the doctor can learn from the courts. I hope the day will come when no psychiatrist will ever say that he is too busy to take the patient's case if it is likely to involve him in legal controversy. I hope the time will come when no psychiatrist will say that he will not become involved with a case unless he can do so as an appointee of the court as opposed to a doctor employed by one of the parties. I hope the doctor will look forward to providing testimony and that some of the thoughts expressed in this discussion will make his day in court a better experience for him.