Purification and infectivity ofEntomophaga grylli (Fresenius) Batko Pathotype 2 againstMelanoplus differentialis (Thomas) [Ort.: Locustidae]

The life stages ofEntomophaga grylli (Fresenius) Batko Pathotype 2 were purified and separated by centrifugation in Percoll^R density-gradient medium. The ranges of buoyant densities for germinated resting spores, germ conidia, and resting spores respectively were: 1.040–1.050, 1.055–1.085, and 1.080–1.120 g/ml. Cuticular invasion by germinated germ conidia was the means by whichMelanoplus grasshoppers became infected. Scanning electron micrographs revealed germination of germ conidia on the visible host integument at 100% RH, but not at 90% RH. Significantly higher mortality (P<0.05) was obtained after 3 weeks with grasshoppers incubated in constant light than in constant dark for 24 h following treatment. The disease was not transmitted by ingestion of any life stage. Contribution N^o 85-153-J, Department of Entomology. Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506.     

Complementarity of genes for resistance to greenbug [Schizaphis graminum (Rondani)], biotype E,in sorghum [Sorghum bicolor (L.) Moench]

Summary Gene complementarity among various sources of resistance to greenbug biotype E was assessed. Analysis of the F₂ generation of crosses between susceptible and resistant parents (mating 1) and among sources of resistance (mating 2) suggested that resistance in sorghum to greenbug biotype E was complexly inherited and, to some extent, dependent on the nature of both the resistant and susceptible parents. Positive transgressive segregation in the F₂ generations of both matings was found to be due to effective plus factors, contributed by both parents in a cross, which complemented each other. The number of plus factors ranged from one to two in the susceptible parents and from two to five in the resistant parents of mating 1, and from one to five in the parents of mating 2. The consistently significant reciprocal effects shown by Sarvasi and PI264453 indicated that these sources had major factors for resistance in their cytoplasms, which were expressed in all their crosses. The results from this study indicated that the sources of resistance complemented each other to give increased number of F₂ segregates with increased resistance. Thus, it should be possible to increase and diversify resistance of sorghum to greenbug biotype E by accumulating different, effective plus factors from various sources through recurrent selection.Contribution No. 90-106-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506, USA     

Identification of two populations of biogenic amine-containing cells in the mouse lung

Summary Two distinct populations of fluorogenic amine-containing cells were observed in the lungs of nine-week old mice: one with an intense yellow emission, which possibly indicates the presence of serotonin; and one emitting a yellow-green fluorescence, which probably indicates the presence of a catecholamine such as dopamine or norepinephrine. Simultaneous identification of two different fluorogenic amine-containing cells, without pre-administration of a precursor to that amine, has not previously been reported. Such evidence of amine-containing cells demonstrated the success of the perfusion-freezing technique and established that cellular storage of fluorogenic amines does occur in vivo under normal physiological conditions. The function of such amine-containing cells has not been established; however, their location and the known physiological effects of amines would suggest regional control of ventilation/perfusion of the lung.The authors wish to thank Dr. Harry Anthony for the use of the fluorescence microscope and Dr. Robert Klemm for use of the photographic facilities. This study was supported by a grant-in-aid from the Kansas Heart Association. Contribution no. 182j, Department of Anatomy and Physiology, KAES, Kansas State University, Manhattan, Kansas 66506     

Regulation of internodal lenght by peroxidase enzymes in grain sorghum

Summary A relationship between height genes (dw locus) and perioxidase was demonstrated by extracting and determining peroxidase specific activity in internode tissue from different height isogenic lines of sorghum Sorghum bicolor (L.) Moench]. Tall plants (2 dwarf) had less peroxidase per gram tissue than their short counterparts (3 dwarf); their F₁ offspring internodes were closer but had more peroxidase than the tall parent. Peroxidase in the F₂ offspring was inversely related to their height and followed a simply-inherited pattern similar to that for height.Among different tissues analyzed, peroxidase concentration in roots was higher than in leaves and internodes, whole internode higher than in pith, and seed embryo higher than in endosperm. Peroxidase activity of nonviable seeds was negligible.Isoelectric focusing provided a more detailed peroxidase zymogram than did gel electrophoresis. Differences in peroxidase bands among tall and short parental plants, F₁ and F₂ segregating groups all appear to be reflected by intensity differences rather than by position or number of bands.Activities of nitrate reductase and acid phosphatase did not correlate with height. That finding provides a control and suggests that peroxidase activity is not associated with height by chance but may have a functional relationship.Contribution no. 1628-j, Dept. of Agronomy and no. 188-j, Dept. of Biochemistry, and no. 962-j, Dept. of Grain Science and Industry, Kansas State University, Kansas Agricultural Experiment Station, Manhattan, Kansas 66506.     

Transfer of Ph I genes promoting homoeologous pairing from Triticum speltoides to common wheat

Diploid-like chromosome pairing in polyploid wheat is controlled by several Ph (pairing homoeologous) genes with major and minor effects. Homoeologous pairing occurs in either the absence of these genes or their inhibition by genes from other species (Ph ^I genes). We transferred Ph ^I genes from Triticum speltoides (syn Aegilops speltoides) to T. aestivum, and on the basis of further analysis it appears that two duplicate and independent Ph ^I genes were transferred. Since Ph ^I genes are epistatic to the Ph genes of wheat, homoeologous pairing between the wheat and alien chromosomes occurs in the F₁ hybrids. Using the Ph ^I gene stock, we could demonstrate homoeologous pairing between the wheat and Haynaldia villosa chromosomes. Since homoeologous pairing occurs in F₁ hybrids and no cytogenetic manipulation is needed, the Ph ^I gene stock may be a versatile tool for effecting rapid and efficient alien genetic transfers to wheat.Contribution no. 93-435-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506-5502, USA     

C-banding variation in the Moroccan oat species Avena agadiriana (2n=4x=28)

The C-banding technique was used to describe the chromosomes of a relatively recently-discovered Moroccan oat species, Avena agadiriana (2n=4x=28). A substantial amount of polymorphism for arm ratios and C-banding patterns was observed among five accessions of this species. However a common set of ten putatively homologous chromosomes was identifiable among the five accessions. The chromosomes of A. Agadiriana do not closely match those of any of the previously described diploid or tetraploid oat species in terms of their arm ratios and C-banding patterns. However, their overall C-banded appearance generally resembles the A/B/D groups of chromosomes of Avena species, rather than the more hetrochromatic C genomes. Implications of these findings in terms of chromosome evolution in the genus Avena are discussed.Contribution no. 95-490-J of the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS, USA     

A survey to determine the occurrence of Histoplasma capsulatum and Cryptococcus neoformans in air-conditioners

Summary A survey of 84 dust samples from 42 air-conditioners and controls was conducted at Kansas State University, Manhattan. Isolations ofC. neoformans andH. capsulatum were attemped using various mycological procedures. H. capsulatum was not recovered in this survey. One isolate ofC. neoformans was obtained. The contaminated air-conditioner yielding the organism contained considerable bird feces, feathers, and dust.Contribution No. 102, Department of Infectious Diseases, Kansas Agricultural Experiment Station, Manhattan, Kansas.     

Intrapulmonary receptors in the bullfrog: Sensitivity to CO2

Summary Single-unit vagal afferent recordings were made on 55 intrapulmonary receptors in 15 anesthetized or decerebrate bullfrogs. Intrapulmonary CO₂ concentration and intrapulmonary pressure were controlled independently by unidirectionally ventilating the lungs. No CO₂ receptors (insensitive to stretch of the lung) of the kind reported in birds and reptiles were found; all 55 receptors were mechano-sensitive. Of these mechanoreceptors, 39 adapted slowly to inflation of the lung and 16 adapted rapidly. Thirtythree of the slowly-adapting receptors and 15 of the rapidly adapting receptors decreased their discharge frequency as intrapulmonary CO₂ concentration was increased. Inflating the lung enhanced CO₂ sensitivity. The results indicate that the frog possesses CO₂-sensitive pulmonary mechanoreceptors similar to those of mammals and reptiles.Abbreviations P _ip intrapulmonary pressure - fractional inspired concentration of CO₂ The authors wish to thank Dalyn Wilson for his help in gathering the experimental animals and for his technical assistance. The study was supported in part by a grant-in-aid from the American Heart Association, Kansas Affiliate, Inc. Contribution No. 78-185-J Department of Anatomy and Physiology, KAES, Kansas State University, Manhattan, Kansas 66506, USA.     

Standard karyotype of Triticum umbellulatum and the characterization of derived chromosome addition and translocation lines in common wheat

A standard karyotype and a generalized idiogram of Triticum umbellulatum (syn. Aegilops umbellulata, 2n = 2x = 14) was established based on C-banding analysis of ten accessions of different geographic origin and individual T. umbellulatum chromosomes in T. aestivum — T. umbellulatum chromosome addition lines. Monosomic (MA) and disomic (DA) T. aestivum — T. umbellulatum chromosome addition lines (DA1U = B, DA2U = D, MA4U = F, DA5U = C, DA6U = A, DA7U = E = G) and telosomic addition lines (DA1US, DA1UL, DA2US, DA2UL, DA4UL, MA5US, (+ iso 5US), DA5UL, DA7US, DA7UL) were analyzed. Line H was established as a disomic addition line for the translocated wheat — T. umbellulatum chromosome T2DS·4US. Radiation-induced wheat — T. umbellulatum translocation lines resistant to leaf rust (Lr9) were identified as T40 = T6BL·6BS-6UL, T41 = T4BL·4BS-6UL, T44 = T2DS·2DL-6UL, T47 = Transfer = T6BS·6BL-6UL and T52 = T7BL·7BS-6UL. Breakpoints and sizes of the transferred T. umbellulatum segments in these translocations were determined by in situ hybridization analysis using total genomic T. umbellulatum DNA as a probeContribution no. 94-349-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506-5502, USA     

Growth of Lactobacillus acidophilus in the absence of folic acid

Soska, Jirí (Kansas State University, Manhattan). Growth of Lactobacillus acidophilus in the absence of folic acid. J. Bacteriol. 91:1840-1847. 1966.-A chemically defined medium, containing no folic acid, was used for the cultivation of Lactobacillus acidophilus R-26. In such a medium, the organism required thymine in addition to a deoxyriboside, purines, pyrimidines, and most amino acids. If thymine was present in this medium, an unlimited exponential growth was possible. The influence of the components of this medium on the growth is described. The concentration and type of adenine compounds in this medium were most important. Adenine and adenosine inhibited utilization of thymine, but not of thymidine, whereas adenylic acid inhibited recovery from amino acid starvation. In the absence of thymine or deoxyribosides, cells continued to grow in length, and after 3 hr a slow decline in viable count ensued.     

Fatal cytauxzoonosis in a free-ranging bobcat (Lynx rufus)

In September 2000, a free-ranging bobcat (Lynx rufus) cub was presented to the Kansas State University Veterinary Teaching Hospital (Manhattan, Kansas, USA) in a moribund state with signs of severe anemia and respiratory difficulty. The cub was euthanized. Gross necropsy findings included multifocal atelectasis, splenomegaly, and pericardial effusion. Microscopic examination revealed subacute pulmonary thrombosis, mild vasculitis in the brain, and large schizont-filled macrophages within blood vessels of all tissues examined. The organisms were typical of the developmental stages of Cytauxzoon felis. Cytauxzoonosis is considered to be a persistent, subclinical infection in the bobcat; however, this cub had lesions consistent with those seen in fatal infections in domestic cats. This case of fatal C. felis infection indicates that some free-ranging bobcats may die of cytauxzoonosis.     

Biochemical characterization of six trisomics of grain sorghum,Sorghum bicolor (L.) Moench

To determine protein differences of grain sorghum disomics and trisomics, we analyzed leaf extracts from six trisomics and a disomic control by disc gel, gel isoelectric focusing, and SDS gel electrophoresis. Based on the number and position of protein bands revealed by Commassie blue staining, the disomic control could be differentiated from the trisomics, and trisomics could be shown to differ among themselves in most cases. SDS gels revealed the most protein bands, followed by isoelectric focusing and disc gel. However, disc gel electrophoresis was the simplest technique of the three and was just as effective in identifying trisomics and differentiating trisomics from the disomic control.Contribution 1596-j, Department of Agronomy, and 182-j, Department of Biochemistry, Kansas State University, Manhattan, Kansas.     

Soybean yield response to residual NO3−N and applied N

Summary During 1976 through 1978, 10N treatments (combinations of N application times and rates) were used in a corn study. Those treatments created different levels of soil NO ₃ ^– –N content that were well-suited to a study of the influence of residual NO ₃ ^– –N and applied N on soybean yield. In April 1979 we applied ammonium nitrate at rates of 0, 75, or 150 kg N/ha to three subplots formed from each of the whole plots (previous N treatment plots). With N fertilization in 1979, seed yield increased where the residual NO ₃ ^– –N amount was less than 190 kg/ha but decreased where the residual amount was greater than 190 kg/ha. As the NO ₃ ^– –N content in the soil increased by 1 kg/ha, the soybean yield increase due to N fertilization in 1979 decreased by approximately 4 kg/ha.Contribution no. 82-368-J, Dep. of Agronomy, Kansas Agric. Exp. Stn., Manhattan, KS 66506, USA     

The distribution of sulfur among various components of the wheat grain

Summary 1. Radioactive sulfur (S³⁵) was introduced into 3 sets of wheat plants grown hydroponically in the greenhouse. When mature, the grain was milled using micromilling procedures.2. Because of the small quantity of samples obtained, a method was developed for concentrating small amounts of sulfate.3. The highest concentration of S³⁵ in the mechanically separated fractions was in the germ, with bran, flour (endosperm), shorts, and red dog containing lesser amounts in that order.4. Radioactivity was approximately the same in the total water-soluble and gluten fractions of flour, although the percentage of nitrogen of total water-solubles was less than gluten.5. Only small differences in count rate were found between the gluten and the water-soluble proteins of the flour, indicating that much of the sulfur radioactivity in the total water-solubles was due to material other than protein.6. S³⁵ in the crude starch was almost completely in the tailings fraction.7. Gliadin was compared with glutenin and found to be somewhat higher in count rate, although nitrogen percentages of both were approximately the same.Contribution No. 538, Department of Botany and Plant Pathology; Kansas Agricultural Experiment Station, Kansas State University, Manhattan. Botany Department serial No. 694.Portion of a dissertation presented as partial fulfillment of the requirement for the Doctor of Philosophy degree in the Department of Botany and Plant Pathology, Kansas State University.     

Calcium transport from Malpighian tubules to puparial cuticle ofMusca autumnalis

Summary The transport of calcium from mineralized granules stored in the Malpighian tubules to the puparium of the face fly,Musca autumnalis De Geer, was studied. Calcium was transported directly from the tubules to the cuticle via the hemolymph. Little, if any, calcium entered the hindgut or other tissues during or prior to transport. A total of approximately 0.8 mg of calcium per larva was transported, beginning at the wandering stage; peak hemolymph concentrations occurred at anterior retraction. Hemolymph calcium levels subsequently decreased as puparial calcium increased. Puparial mineralization utilized most of the minerals stored during the larval stage, with lesser amounts of minerals being recovered in the adult or excreted. Deposition of mineral salts in the cuticle was accompanied by an increase in cuticular pH from 7.0 to 8.4. The house fly,Musca domestica L., which contains much lower concentrations of minerals in the puparial cuticle, exhibited no increase in cuticular pH during pupariation. Biomineralization of the face fly puparial cuticle appears to occur, in part, as a result of ionic equilibria involving calcium and magnesium phosphates and carbonates, which have relatively low solubility products at alkaline pH.Contribution No. 87-237-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, Kansas. Cooperative investigation between Agricultural Research Service, U.S. Department of Agriculture and the Kansas Agricultural Experiment Station. K.J.K. is a research chemist and adjunct professor at U.S. Grain Marketing Research Laboratory and Kansas State University, respectively. Mention of a proprietary product in this paper does not imply its approval by the USDA to the exclusion of other products that may also be suitable. Reprint requests to A.B. Broce     

Effectiveness of ph gene in inducing homoeologous chromosome pairing in agrotricum

Summary We observed pairing, when the ph gene was present, between wheat (Triticum aestivum L. em. Thell.) chromosome 4B, and an Agropyron intermedium (Host) Beauv. chromosome (Ai) carrying a gene resistant to wheat streak mosaic (WSM). In a monosomic addition polyhaploid [2n = 22 = 19' + 5B' (ph) + 4B' + Ai'], we recorded an average of 4.1 bivalents and 0.3 trivalents per cell. Induced homoeologous pairing was most effective when both 5B chromosomes carrying ph gene were present. Our data suggest that chromosome 4B of wheat and the Agropyron chromosome (Ai) carrying a gene for resistance to WSM are homoeologous and that it is possible to use either ph mutant or nullisomic 5B stock to induce genetic recombination between the two chromosomes.Contribution No. 1657-j, Kansas State Agric. Expt. Sta., Manhattan, KS. The research is partially supported by a grant from Kansas Wheat Commission     

The effect of parental divergence on F2 heterosis in winter wheat crosses

Summary In winter wheat (Triticum aestivum L.), the development of a methodology to estimate genetic divergence between parental lines, when combined with knowledge of parental performance, could be beneficial in the prediction of bulk progeny performance. The objective of this study was to relate F₂ heterosis for grain yield and its components in 116 crosses to two independent estimates of genetic divergence among 28 parental genotypes of diverse origins. Genetic divergence between parents was estimated from (a) pedigree relationships (coefficients of kinship) determined without experimentation, and (b) quantitative traits measured in two years of field experimentation in Kansas and North Carolina, USA. These distances, designated (1 -r) and G, respectively, provided ample differentiation among the parents. The 116 F₂ bulks were evaluated at four locations in Kansas and North Carolina in one year. Significant rank correlations of 0.46 (P = 0.01) and 0.44 (P = 0.01) were observed between G and grain yield and kernel number heterosis, respectively. Although (1 -r) was poorly associated with grain yield heterosis, G and midparent performance combined to account for 50% of the variation in F₂ yields among crosses when (1 -r) was above the median value, whereas they accounted for only 9% of the variation among crosses when (1-r) was below the median. Midparent and (1 -r) had equal effects on F₂ grain yield (R ²= 0.40) when G was greater than the median value. A breeding strategy is proposed whereby parents are first selected on the basis of performance per se and, subsequently, crosses are made between genetically divergent parents that have both large quantitative (G) and pedigree divergence (1 -r).Paper No. 12162 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7643, and Contribution No. 89-396-J of the Kansas Agricultural Experiment Station, Manhattan, KS 66506     

Induction of Cold Hardiness by Salt Stress Involves Synthesis of Cold- and Abscisic Acid-Responsive Proteins in Potato (Solanum commersonii Dun)

Salt stress has been found to increase frost tolerance in someherbaceous species. In an attempt to understand the molecularbasis of the frost tolerance induced by salt stress, the effectof salt (100 mM NaCl) on total proteins in stem-cultured potatoplantlets (Solarium commersonn Dun) was analyzed on two-dimensionalgels. Nine salt-induced proteins were identified after 24 hsalt treatment, at which time cold hardiness increased by threedegrees. Direct comparisons of the proteins with those inducedby cold- and abscisic acid(ABA)-treatments revealed that fiveof the salt-induced proteins were also induced by cold(4°/2°C)-treatmentand seven were also induced by ABA(40µM)-treatment. Threeproteins (M_rr/pls 13/7.0, 27/6.6 and 48/6.9) were induciblein both cold- and ABA-treatments in association with frost hardening.After 6 h salt treatment, endogenous ABA levels in plantletleaves showed a transient six-fold increase before cold hardinessdeveloped. The results suggest that salt-induction of cold hardinessinvolves the synthesis of cold and ABA-responsive proteins andthe alteration of protein synthesis is mediated by ABA elevatedupon salt stress. This study also suggests that a subset ofproteins induced by cold and ABA-treatments are related to saltstress. ¹Scientific Journal Series Paper No. 20787 of the MinnesotaAgricultural Experimental Station, St. Paul, MN 55108, U.S.A. ²Present address: Department of Biochemistry Willard Hall, KansasState University, Manhattan, KS 66506, U.S.A. ³Present address: Via G.B. Marino 13, 80125-Napoli, Italy ⁴Present address: Institute of Biological Chemistry, WashingtonState Univ., Pullman, WA 99164, U.S.A.     

Standard karyotype of Triticum searsii and its relationship with other S-genome species and common wheat

C-banding polymorphism was analyzed in 14 accessions of Triticum searsii from Israel, and a generalized idiogram of the species was established. One accession was homozygous for whole arm translocations T1S^sS·4S^sS and T1S^sL·4S^sL. C-banding analysis was also used to identify 7 T. aestivum cv Chinese Spring-T. searsii disomic chromosome addition lines, 14 ditelosomic chromosome addition lines, 21 disomic whole chromosome, and 31 ditelosomic chromosome substitution lines. The identity of these lines was further confirmed by meiotic pairing analysis. Sporophytic and gametophytic compensation tests were used to determine the homoeologous relationships of the T. searsii chromosomes. The results show that the T. searsii chromosomes do not compensate well for their wheat homoeologues. The C-banding patterns of T. searsii chromosomes are distinct from those of other S-genome species and from the B-genome chromosomes of wheat, indicating that T. searsii is not a direct B-genome donor species of T. turgidum and T. aestivum.Contribution No. 95-72-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, Kansas, USA     

High transmission of paternal plastid DNA in alfalfa plants demonstrated by restriction fragment polymorphic analysis

Summary A high frequency of paternal plastid transmission occurred in progeny from crosses among normal green alfalfa plants. Plastid transmission was analyzed by hybridization of radiolabeled alfalfa plastid DNA (cpDNA) probes to Southern blots of restriction digests of the progeny DNA. Each probe revealed a specific polymorphism differentiating the parental plastid genomes. Of 212 progeny, 34 were heteroplastidic, with their cpDNAs ranging from predominantly paternal to predominantly maternal. Regrowth of shoots from heteroplasmic plants following removal of top growth revealed the persistence of mixed plastids in a given plant. However, different shoots within a green heteroplasmic plant exhibited paternal, maternal, or mixed cpDNAs. Evidence of maternal nuclear genomic influence on the frequency of paternal plastid transmission was observed in some reciprocal crosses. A few tetraploid F₁ progeny were obtained from tetraploid (2n=4x=32) Medicago sativa ssp. sativa x diploid (2n=2x=16) M. sativa ssp. falcata crosses, and resulted from unreduced gametes. Here more than the maternal genome alone apparently functioned in controlling plastid transmission. Considering all crosses, only 5 of 212 progeny cpDNAs lacked evidence of a definitive paternal plastid fragment.Contribution No. 89-524-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan