Unexpected progeny obtained from glasshouse study of soil pH influence onAcaulospora laevis inoculum

Summary Potential growth stimulation of two hosts by acid-tolerantAcaulospora laevis was tested in a soil adjusted to 5 pH levels from 5.0 to 7.7. By wet-sieving methods, the field-source inoculum was essentially a pure culture ofA. laevis spores. Upon harvest of sweetgum a completely different spore-type was found in large quantities;A. laevis spores were relatively few to non-existent. The results dramatically underscore the need for caution against relying solely on pre-trial identifications of inoculum spores or presuming that apparently single-species cultures/inoculum will remain pure throughout an experiment.Contribution of the Oregon State University Agricultural Experiment Station in cooperation with the US Department of Agriculture, Agricultural Research Service, technical paper #7366 of the former.     

Evidence for the presence of proteinase inhibitor I in vacuolar protein bodies of plant cells

Summary Protein bodies induced in tomato leaf cells by wounding were shown to contain proteinase Inhibitor I by using ferritin-labelled antibodies, fluorescein-labelled antibodies, and cytochrome C-labelled antibody fragments. Both pre-embedding and postembedding techniques were used. Nonspecific binding was least when p-formaldehyde was used as the initial fixative followed by treatment with cytochrome c-labelled antibody fragments.Abbreviations Fab antibody fragments - BSA bovine serum albumin - GMA glycol methacrylate - THB Tris-HCl buffer Taken in part from a doctoral (Ph.D.) dissertation submitted to Washington State University by Vivian V. Yang. This work was supported largely by NSF Grant GB-29614X (LKS) and in part by the United States Department of Agricultural Cooperative States Research Service Grant 316-15-30 (CAR), the National Science Foundation Grant GB-37972 (CAR), and the College of Agriculture Research Center, Washington State University, Pullman, WA 99163, Scientific Paper No. 4525, Project 1791.Program in Genetics and Department of Botany. To whom reprint requests should be sent.Department of Agricultural Chemistry and Program in Biochemistry and Biophysics.     

Differential analysis of sulfhydryl and disulfide groups of intact spores

Blankenship, L. C. (Eastern Utilization Research and Development Division, Washington D.C.), and M. J. Pallansch. Differential analysis of sulfhydryl and disulfide groups of intact spores. J. Bacteriol. 92:1615-1617. 1966.-Fluorescence quenching of fluorescein mercuric acetate in alkaline medium (1 n NaOH) was found to be an accurate, sensitive method for differential analysis of sulfhydryl and disulfide groups of intact bacterial spores. Disulfide content of three species of spores was found to be slightly higher than reported by other workers. A significant increase in sulfhydryl group concentration was observed after physiological germination, indicating participation of a disulfide-reducing system in the germination process.     

Mitochondrial DNA synthesis during fungal spore germination

Summary Germinating spores of the fungus Botryodiplodia theobromae incorporated guanine-8-C¹⁴ into both the nuclear DNA and mitochondrial DNA fractions. Ethidium bromide inhibited the synthesis of mitochondrial DNA without having a significant effect on nuclear DNA synthesis or on the rate and extent of spore germination. Rates of leucine and uracil incorporation and of oxygen uptake were not significantly affected by ethidium bromide until germination was nearly completed. Mitochondrial DNA synthesis is apparently not required for germination of the spores of B. theobromae but is probably essential to continued vegetative growth.Abbreviations DNA deoxyribonucleic acid - mit-DNA mitochondrial DNA - nuc-DNA nuclear DNA - RNA ribonucleic acid - EB ethidium bromide - Tris tris (hydroxymethyl)aminomethane Published with the approval of the Director as Paper No. 3331, Journal Series, Nebraska Agricultural Experiment Station. Research reported was conducted under Project No. 21-17. Paper No. 7877, Scientific Journal Series, Minnesota Agricultural Experiment Station.     

Physiological and chemical studies on mould proteins

Summary A series of experiments was done to determine the best conditions for the extraction of protein from mould spores (Penicillium roqueforti). These experiments included studies on the influence of the extraction technique, the nature of the extracting fluid, spores glass: beads: extracting fluid ratio, and the time of extraction.Some work was done in fractionating the extractable nitrogenous substances and proteins of the spores. It was found that the extracted nitrogen could be fractionated into 12% dialysable low molecular weight nitrogen compounds and 88% crude protein, the latter was made up of about 40% albumens and 60% globulins.Visiting Research Professor, on scientific leave from National Research Center, Cairo, U.A.R.; Appointment supported by the International Cooperation Administration under the Visiting Research Scientists Program administered by the National Academy of Sciences of the United States of America. The work also was supported in part by grant E-1201 from the National Institutes of Health.     

Interspecific hybrids between kenaf ( Hibiscus cannabinus ) and roselle ( H. sabdariffa )

The successful interspecific cross is reported for the first time between kenaf (Hibiscus cannabinus L.), a diploid species (2n=36) and roselle (Hibiscus sabdariffa L.), a tetraploid species (2n=72). Kenaf, grown for its bast fiber and also under investigation as a source of paper pulp, is fast-growing and well adapted to mechanical harvesting, but susceptible to root-knot nematodes. Roselle, also grown for its bast fiber, is slower growing, not well adapted to mechanical harvesting, but certain varieties are resistant to root-knot nematodes. Five hybrid plants were produced from the pollination of 4,445 flowers of kenaf with pollen from roselle; no hybrid plants were produced from 2,655 pollinations made in the reciprocal direction. One line of roselle was the parent of 3 of the 5 hybrids; one line of kenaf was the parent of 2 of these 3. The F₁ hybrids were triploid, and varied in vigor, growth habit and vegetative morphology, but had similar flowers. Two of the F₁ hybrids showed high pollen fertility, apparently as a result of restitution at first meiotic division leading to unreduced spores. These two hybrids each produced a small amount of seed, which gave rise to an F₂ population of 22 plants. The F₂ plants vary in vigor but are morphologically uniform, have thick leaves with mosaic sectors, and are presumably spontaneous allohexaploids. The theoretical possibilities of increasing the percentage of recovery of the F₁ interspecific hybrids and of developing a synthesized hybrid variety useful for bast fiber and paper pulp are discussed. Research done cooperatively by Crops Research Division and Agricultural Engineering Research Division, Agricultural Research Service, U.S. Department of Agriculture (USDA), and the University of Florida Agricultural Experiment Station. Cytological investigations were carried out at the Department of Biological Sciences, Florida State University. Thanks are extended to the many individuals and organizations who supplied material for this study, and to M. Griffin Bell and E. Otho Richey, Agricultural Research Technicians, for their enthusiastic assistance.     

A computer-assisted examination of the storage protein genetic variation in 841 accessions of Triticum dicoccoides

Summary Triticum turgidum L. var. dicoccoides (wild emmer) is an important genetic resource for increasing the protein content of common wheat (Triticum aestivum L.). Many studies have shown that the presence or absence of bands in sodium dodecyl sulfate polyacrylamide (SDS-PAGE) electrophoregrams of wheat storage proteins to be of a purely genetic character. A total protein extraction and SDS-PAGE technique was used to estimate the storage protein genetic variability among 841 accessions of wild emmer collected from various ecological regions in the Middle East. In addition, a computer data bank was developed, recording the onedimension electrophoregram bands for each accession by molecular weight (MW) and relative Coomassie Blue staining intensity as determined from densitometer scans. Analyses of this information are being used to identify specific accessions for further study by two dimension electrofocusing-electrophoresis and breeding and genetic analyses. The computer-assisted analyses indicated that the greatest genetic variability occurs for proteins in the high MW region (above 70,000 MW) followed by those in the medium range (70,000 to 33,300 MW). Comparatively little variability was revealed for protein subunits of below 33,300 MW.Scientific Paper No. 6591. College of Agriculture Research Center, Washington State University, Pullman, Project No. 1568. This work was supported in part by the U.S.-Israel Binational Agricultural Research and Development Fund (BARD)     

Morphology of the spore of some strains of Clostridium botulinum type E

Hodgkiss, W. (Torry Research Station, Aberdeen, Scotland), and Z. John Ordal. Morphology of the spore of some strains of Clostridium botulinum type E. J. Bacteriol. 91:2031-2036. 1966.-The spores of four strains of C. botulinum type E show an unusual and elaborate morphology. Numerous tubular appendages radiate from the surface of the spore. The spore and its appendages are enclosed in a delicate exosporium. The electron microscopic morphology of the spores, as seen in metal-shadowed and negatively stained preparations, and by the carbon-replica technique, is described.     

Bacillus anthracis contamination and inhalational anthrax in a mail processing and distribution center

AIMS: Four inhalational anthrax cases occurred in a large mail processing and distribution center in Washington, DC, after envelopes containing Bacillus anthracis spores were processed. This report describes the results of sampling for B. anthracis spores during investigations conducted in October and December 2001. METHODS AND RESULTS: Wet swabs, wet wipes, vacuum sock, and air-filter samples were collected throughout the facility to characterize the extent of building contamination. The results showed widespread contamination of B. anthracis spores, particularly associated with one delivery bar code sorter (DBCS) machine that had sorted the spore-containing envelopes and an area where the envelopes were handled by postal workers. Spore concentrations decreased as distance from the DBCS machine increased, but spores were widely dispersed into surrounding areas. CONCLUSION: The spatial distribution of culture positive samples was closely related to the work areas of the inhalational anthrax cases and supported epidemiological evidence that the workers became ill from exposure to B. anthracis spores in areas where the contaminated envelopes had travelled. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this investigation were used to guide decontamination efforts and provided baseline spore concentrations for follow-up measurements after the building had been cleaned. Implementing methods to reduce aerosolization and dispersion of dust within the facility would reduce postal workers' potential exposures to bioterrorism agents.     

First Report of Anthracnose of Ledebouriella seseloides Caused by Colletotrichum gloeosporioides in Korea

In 2012, dark brown spots were observed on leaves of Ledebouriella seseloides (Fang Feng) in several research plots located at the Goseong Agricultural Research Extension services in Gyeongam Province, Republic of Korea. A fungus was isolated from the infected plants which produced pink‐coloured spores in mucilage on PDA and conidial morphology suggested that the causal agent was Colletotrichum gloeosporioides. Internal transcribed spacer sequences of the pathogen showed 99% identity to those of C. gloeosporioides. Pathogenicity of the isolate was proved by Koch's postulates. This is the first report of anthracnose in L. seseloides caused by C. gloeosporioides.     

Q & A. [interview

Elaine and Gary Ostrander spent their youth in New Jersey and New York before heading to Nebraska for their teen years and eventually Washington State for High School and college, as their father moved around in library administration. Elaine was an undergraduate at the University of Washington, a graduate student at the Oregon Health Sciences University and a postdoc with James Wang at Harvard, studying DNA supercoiling. She next went to Berkeley, where she began the canine genome project, initiating the meiotic linkage map and working on human chromosome 21 at the Lawrence Berkeley National Labs. In 1993 she moved to the Fred Hutchinson Cancer Research Center where she is now a Member of the Divisions of Clinical Research and Human Biology. She is also an Affiliate Professor of Genome Sciences and Biology at the University of Washington, and heads the Program in Genetics at the Hutchinson Center. Gary completed his undergraduate degree in Biology at Seattle University, a M.S. degree at Illinois State University and a Ph.D at the University of Washington in Ocean and Fisheries Science. He went on to be a postdoc in the Department of Pathology at the University of Washington Medical School while being mentored by Senitroh Hakomori of the Fred Hutchinson Cancer Research Center and Eric Holmes of the Pacific Northwest Research Foundation. His work focused on using novel aspects of the biology of fishes to address fundamental questions about cancer. He subsequently held both faculty and administrative positions at Oklahoma State University. Since 1996, he has been at the Johns Hopkins University, where he currently holds academic appointments in the Departments of Biology and Comparative Medicine and is the Associate Provost for Research.     

The use of X-ray microanalysis to demonstrate the uptake of the molluscicide copper sulphate by slug eggs

Summary Copper has been localized in copper sulphate treated eggs of the slug Agriolimax reticulatus (Mü). This has been accomplished using a freeze-fracture technique, the freeze-dried halves of the fractured eggs being analysed in the scanning electron microscope, using both energy dispersive and wavelength dispersive X-ray microanalysis systems. The distribution of copper obtained using these methods has been compared with that achieved using a standard histochemical technique. Both techniques revealed that the copper is initially retained in the perivitelline membranes. The application of X-ray microanalysis to such studies is discussed.This research was supported by the Agricultural Research Council (G.B.), Grant No. AG 72/13     

Penicillium urticae Bainier enumeration in soils

Summary A dilution plating method estimatedPenicillium urticae Bainier numbers in soil. This method, which used an agar layering technique and a cyclic incubation of 8 hours at room temperature (about 25°C) and 16 hours at 5°C, permitted the differential growth and sporulation in favor ofP. urticae B. over other common soil fungi.Procedures of extraction, paper chromatography, infrared analysis, and bioassay assayed for accumulated patulin. A combination of these methods routinely estimatedP. urticae B. numbers in soils and authenticated patulin production by selected isolates.Contribution from the Northern Plains Branch, Soil and Water Conservation Research Division, Agricultural Research Service, USDA, in cooperation with the Nebraska Agricultural Experiment Station, Lincoln. Published as Paper No. 2275, Journal Series, Nebraska Agricultural Experiment Station.Soil Scientist, USDA, Grand Junction, Colorado (formerly Chemist, USDA, Lincoln, Nebraska); and Microbiologist, USDA, Lincoln, Nebraska, respectively.     

Vacuolar protein bodies in tomato leaf cells and their relationship to storage of chymotrypsin inhibitor I protein

Summary Electron microscopy of leaves of tomato has shown that tissue containing chymotrypsin inhibitor I protein has protein in the cell vacuoles. The vacuolar protein was found either as many small bodies or as few large bodies. The data indicate that the vacuole is a temporary storage site for protein which may play an important role in growth and development of the plant. This strongly suggests that the plant-cell vacuole is something more than a site for terminal deposition of waste products. The system offers an unusual opportunity to study the biochemistry and ultrastructure of synthesis, vacuolar deposition, and recall of a well-characterized plant protein.This work was supported in part by Washington State Medical and Biological funds, U.S. Public Health Service Grants 2K3GM17059 and GM12505, and by U.S.D.A. C.S.R.S. Grant No. 915-15-29. College of Agriculture scientific paper 3442, Projects 1791 and 1996.Program in Genetics and Department of Botany.Department of Agricultural Chemistry.Department of Agricultural Chemistry. Research Career Development Awardee of the U.S. Public Health Service.     

Enzymes involved in the utilization of carbohydrates by two strains of myxobacteria

Summary Extracts prepared from two strains of myxobacteria, grown with glucose as the primary carbon source, have been shown to contain a hexokinase that phosphorylates glucose to a greater extent than fructose. Phosphohexose isomerase, 6-phosphofructokinase, aldolase and a DPN-specific triose phosphate dehydrogenase are also present. In addition a TPN-specific glucose-6-phosphate dehydrogenase and a system catalyzing the reduction of DPN in the presence of 6-phosphogluconate have also been demonstrated.Supported in part by State of Washington funds for medical and biological research, and in part by the Office of Naval Research under Contract N80nr-520/III with the University of Washington.     

花椰菜BobACT基因的克隆及其作为内参基因的研究

实时荧光定量PCR技术是探索植物基因功能和调节机理的有效手段。选择合适的内参基因是获得实时荧光定量PCR准确性数据的必备条件。ACT基因高度保守且表达稳定,常作为内参基因被广泛应用。为了获得花椰菜ACT基因,以转录组测序和RT-PCR方法为手段克隆得到花椰菜肌动蛋白基因Actin。该基因等电点为5.395,理论分子量为41.77 kD;其cDNA开放阅读框长1134 bp,编码氨基酸377个,GenBank登录号为MG598643。Wolf Psort分析发现,BobActin蛋白亚细胞定位于细胞质基质中。Motif Scan分析显示,BobActin蛋白质的氨基酸序列4～377位为Actin保守结构域。进化分析表明,同源序列基因编码的蛋白质与同为十字花科的甘蓝、芜菁和油菜同源蛋白的相似性达到90%以上,具有高度的保守性。在此基础上,设计了1对荧光定量PCR引物,分析显示,该引物具有较高的特异性和扩增效率,在花椰菜根、茎、花、花球、叶片等不同组织和低温、高温、盐处理、干旱处理、ABA处理等胁迫处理下均能稳定表达,适合在花椰菜基因表达研究中作为内参基因,为开展花椰菜重要功能基因的挖掘、表达模式以及调控机理的研究提供参考。花椰菜在内参基因方面的研究还处于初步阶段,今后可继续克隆其他内参基因,丰富花椰菜的内参基因库,从而进一步提高花椰菜基因表达分析研究的稳定性、重复性和准确性。     

The effect of available carbon and nitrogen in straw on soil and ash aggregation and acetic acid production

Summary Aerobically decomposed straws containing various contents of available C and N were tested for resultant aggregating effect on Mt. St. Helen's ash and Palouse silt loam. Aggregation decreased when straw N content increased in the range 0.25–1.09% w/w. These results suggest that microbial extra-cellular products are very important for stabilizing soils. Microbial production of acetic acid, which can be phytotoxic to wheat plant seedlings, was greatest initially from the 1.09% N w/w straw. After the first three days of aerobic decomposition, acetic acid production was not linked to the straw N content. The potential of barley and wheat straw to serve as a substrate for acetic acid production was greater than that of the remains of the flowering heads (chaff). However, the chaff might pack more tightly than the straw in the field, which would increase effectively its acetic acid concentration over that of the straw. Contribution from Agric. Res. Serv., U.S. Dep. of Agric., in cooperation with the College of Agric. Res. Center, Washington State Univ., Pullman, WA 99164; and Agricultural Research Council, Letcombe Laboratory, Wantage, Oxon, Great Britain. WSU Scientific Paper No. 6556. Research was conducted at Letcombe Laboratory.     

Soil water potential as a factor in the ecology ofFusarium roseum f. sp.cerealis ‘Culmorum’

Summary The soil water potential (inferred from vapor pressure measurements by thermocouple psychrometry) influenced both chlamydospore germination and continuing growth of germlings ofFusarium roseum f. sp.cerealis ‘Culmorum’ the same way in two different soils. Chlamydospore germination in both Ritzville silt loam (RSL) and Palouse silt loam (PSL) amended with about 2,500 ppm C (as glucose) and 250 ppm N (as ammonium sulfate) was 40–50 per cent in 24 hours at water potentials down to −50 to −60 bars. Some germination occurred by 72 hours at −80 to −85 bars in both soils but not at lower potentials. At a potential of −10 bars or higher, germ tubes lysed or converted into new chlamydospores within 48–72 hours after germination, whereas at lower potentials germlings branched and appeared to grow for at least 6 days. Bacterial numbers/g of RSL, 24 and 72 hours after adding nutrients, were 200 to 300 times greater in soil at water potentials of −5 bars or more than in comparably treated soil at about −14 to −17 bars or less. Markedly reduced bacterial activity appeared to coincide with a water potential of about −9 to −10 bars. When streptomycin and neomycin (300 ppm each) were mixed into the soil in addition to nutrients, the survival of germlings of Culmorum was greatly enhanced, even in soil at potentials of less than −1 bar. Indications were that soil water potentials of −10 bars or more favored bacterial activity, and that this in turn repressed growth of germlings of Culmorum. Culmorum infections of below-ground parts of wheat are serious primarily in drier soils, possibly because the fungus escapes bacterial antagonism but can still extract water for growth. Cooperative investigations, Crops Research and the Water and Soil Conservation Research Divisions, Agricultural Research Service, U.S. Department of Agriculture and the Agricultural Experiment Stations of Idaho, Montana, Oregon, Utah, and Washington. Scientific Paper No.3152, College of Agriculture, Washington State University, Pullman.     

Plant growth-inhibitory pseudomonads colonizing winter wheat (Triticum aestivum L.) roots

Summary Pseudomonads, which inhibit root extension, can be present in the winter wheat rhizosphere in large numbers, but they are not detectable until late winter or early spring. Their presence was not related to the presence of wheat straw residues or type of tillage, although they were present on the wheat residues when they appeared in the rhizosphere. Wheat seedlings were more sensitive to the bacteria at 15° C than at 20° C during bioassays. The type of agar used in the bioassay can affect the results obtained. The inhibitory factor expressed by the pseudomonads is quite variable and is radically affected by transfer of isolates.Contribution from Agric. Res. Serv. U.S. Dept. of Agric., in cooperation with the College of Agric. Res. Center. Washington State Univ., Pullman, WA 99164 (Scientific Paper No. 6743); and Agricultural Research Council, Letcombe Laboratory, Wantage, Oxon OX12 9JT, England.     

Calcium and inositol trisphosphate receptors

Work from the authors' laboratory is supported by the Wellcome Trust, and the Medical, and Agricultural and Food Research Councils. CWT is a Lister Institute Research Fellow.