Effect of gibberellic acid treatment,and nutrient supply through detached tillers,upon haploid frequency in barley

Summary Haploids from Hordeum vulgare (2n = 14) X H. Bulbosum (2n = 14) crosses result after fertilization from the subsequent elimination of bulbosum chromosomes during early embryo development. Seed set from the cross is high but embryo culture is necessary to obtain seedlings. Application of gibberellin A₃ (GA₃) to pollinated florets was effective for increasing the frequency of haploid seedlings produced on both nutrient-fed detached tillers and intact plants. GA₃ increased both seed set and embryo yield. The number of cells per embryo during its transition to the haploid state was increased two to three times following GA₃ treatments. Enhanced embryo and endosperm development was attributed to increased mitotic activity. The number of visibly differentiated embryos was doubled to about 35 % of the cultured embryos after GA₃ was applied to detached tillers in nutrient solution. About 70 % of the resulting haploid plants developed from the visibly differentiated embryos. The detached tiller technique offers a convenient method of culturing haploids from field-grown plants.     

Parthenogenetic development of bovine oocytes treated with ethanol and cytochalasin B after in vitro maturation.

The present study was conducted to investigate the effects of different culture durations (24-36 hr) on bovine oocyte maturation in vitro and the effect of the presence or absence of cumulus cells at the time of treatment to induce parthenogenetic activation (exposure to ethanol and cytochalasin B; CB) (experiment I). The effects of dosage (2.5 or 5.0 micrograms/ml) and incubation time (2.5, 5, or 10 hr) in CB (experiment II) on the subsequent development to the blastocyst stage in vitro was also investigated. In experiment I, cleavage and development to the blastocyst stage were not affected by the presence or absence of cumulus cells at the time of parthenogenetic activation. However, the 24-hr culture duration for in vitro maturation had a significantly lower rate of development to the blastocyst stage than the longer culture durations (27-36 hr). In experiment II, treatment with 5 micrograms/ml CB for 5 hr showed the highest percentage of development to blastocyst in the oocytes matured for both 27 and 30 hr. To determine the viability of the parthenogenetic embryos (morulae and blastocysts), four recipient heifers received two embryos each, and one heifer was found to be pregnant on day 35 following transfer. Although fetal heartbeat was not observed, the subsequent estrus was prolonged in all heifers. The present results demonstrate development of in vitro-matured, parthenogenetically activated bovine embryos up to the preimplantation stage.     

Increasing carbon dioxide from five percent to ten percent improves rabbit blastocyst development from cultured zygotes.

One-cell rabbit zygotes were cultured at 39 degrees C in basal synthetic medium II (BSM-II) with 5%, 10%, or 15% CO2 and humidified air to determine the effect of CO2 concentration on development in vitro. After 4 days in culture, 37% of the embryos grown in 10% or 15% CO2 had reached the hatching blastocyst stage, but only 10% of the embryos were hatching when cultured under 5% CO2 (P = 0.01). Over all blastocysts, cell numbers were 207, 246, and 205 for the 5%, 10%, and 15% CO2 treatments, respectively. In a second experiment to determine if there was a beneficial effect, particularly at the blastocyst stage, of a higher concentration of CO2, embryos were cultured 4 days in either 5% or 10% CO2 or for 2 days in 5% CO2 followed by 2 days in 10% CO2. The numbers of blastomeres per embryo and embryo diameter were greater (P < 0.05) in embryos cultured continuously in 10% CO2 or in 10% CO2 only during days 3 and 4 of culture than in embryos cultured continuously in 5% CO2. In a third experiment, one-cell rabbit zygotes were cultured with 5% or 10% CO2 in a defined, protein-free medium consisting of 1:1 RPMI 1640 and Dulbecco's modified Eagle's medium. The proportion of embryos hatching and cell counts were significantly greater (P < 0.01) when cultured in the presence of 10% CO2. These data indicate that a 10% CO2 atmosphere exerts a beneficial effect on the development of zygotes into expanding and hatching rabbit blastocysts in vitro.     

Ecological release in feeding behaviour: the case of bluegills in Japan

The stomach contents were analyzed monthly for each year-class to elucidate the foraging pattern of bluegills in a small vegetated lake by the frequency occurrence and the points methods. Seasonal dietary changes of the year-classes were considered comparing the monthly fluctuations in abundance of major prey organisms. Though these bluegills are dietary generalists and opportunists like those in North America, their foraging pattern was characterized by a relatively clearer dietary shift during ontogeny and a wider food niche including piscivorous than those of bluegills with congeners in their home land. Therefore this finding provides evidence of the ecological release caused by the absence of congeners.     

Root biomass fraction as a function of growth degree days in wheat

Root, underground and above-ground biomass were measured on various wheat cultivars from 1986 to 1988 in the south-east of France. The results are expressed as root: total (f _r) or underground: total (f _u) biomass fractions. Observed f _r and f _u values are in good agreement with previous results. f _r and f _u decrease steadily from emergence to maturity, with an exponential tendency. When using cumulative growth degree days since emergence relative to cumulative growth degree days until ear emergence () as time scale, f _r and f _u can be expressed as simple functions of % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGceaqabeaacaWGMb% addaWgaaqaaiaadkhaaeqaamaabmaabaGccqaH4oqCdaahaaWcbeqa% aiaacQcaaaaamiaawIcacaGLPaaakiabg2da9iaaicdacaGGUaGaaG% imaiaaiwdacqGHRaWkcaaIWaGaaiOlaiaaiwdacaaI4aGaamyzamaa% CaaaleqabaGaeyOeI0IaaGymaiaac6cacaaI0aGaaGioaiabeI7aXn% aaCaaameqabaGaaiOkaaaaaaaakeaacaWGMbaddaWgaaqaaiaadwha% aeqaamaabmaabaGccqaH4oqCdaahaaWcbeqaaiaacQcaaaaamiaawI% cacaGLPaaakiabg2da9iaaicdacaGGUaGaaGymaiaaikdacqGHRaWk% caaIWaGaaiOlaiaaiIdacaaI4aGaamyzamaaCaaaleqabaGaeyOeI0% IaaGOmaiaac6cacaaIYaGaaGioaiabeI7aXnaaCaaameqabaGaaiOk% aaaaaaaaaaa!610D!\[\begin{gathered} f_r \left( {\theta ^* } \right) = 0.05 + 0.58e^{ - 1.48\theta ^* } \hfill \\ f_u \left( {\theta ^* } \right) = 0.12 + 0.88e^{ - 2.28\theta ^* } \hfill \\ \end{gathered} \]The incremental root biomass partitioning coefficient, % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaeqySde2aaS% baaSqaaiaadkhaaeqaaOGaeyypa0JaaiikaiaadsgacaWGxbWaaSba% aSqaaiaadkhaaeqaaOGaai4laiaadsgacaWG0bGaaiykaiaac+caca% GGOaGaamizaiaadEfadaWgaaWcbaGaamiDaaqabaGccaGGVaGaamiz% aiaadshacaGGPaaaaa!4834!\[\alpha _r = (dW_r /dt)/(dW_t /dt)\], which describes the net increase in root biomass dW _r over time dt relative to the increase in total biomass (dW _r) over the same time period, has been derived from f and the relative growth rate. Its time course is accurately represented by% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaeqySdegdda% WgaaqaaiaadkhaaeqaamaabmaabaGccqaH4oqCdaahaaWcbeqaaiaa% cQcaaaaamiaawIcacaGLPaaakiabg2da9iabgkHiTiaaicdacaGGUa% GaaGymaiaaiwdacqGHRaWkcaaIWaGaaiOlaiaaiAdacaaIZaGaamyz% amaaCaaaleqabaGaeyOeI0IaaGimaiaac6cacaaI5aGaaGioaiabeI% 7aXnaaCaaameqabaGaaiOkaaaaaaaaaa!4D15!\[\alpha _r \left( {\theta ^* } \right) = - 0.15 + 0.63e^{ - 0.98\theta ^* } \]Under our experimental conditions, with no severe water stresses or nutrient deficiencies, and for our sampling frequency, around 2 weeks, the development scale , is the main factor governing the time courses of f _r, f _u and _r.     

A study of factors affecting anther culture of cauliflower (Brassica oleracea var. botrytis)

Experiments on three autumn-heading cauliflower genotypes (2 hybrids and a genotype selected from a population) were conducted to study different factors affecting anther culture. Culture conditions of the donor plants proved to be important: the best results were obtained during spring in a greenhouse where the temperature was maintained between 10 and 20°C. Overall winter and spring seemed more suitable than summer and early autumn for culture establishment. The optimal bud development stage depended on the genotype: for the hybrid 702, the greatest number of embryos for 100 plated anthers was obtained at the uninucleate pollen stage of the microspores; for V23.2 and 703, the optimal stage of the buds corresponded to the first mitotic division. Sucrose proved to be the best carbon supply for embryogenesis with an optimal concentration of 140 g l^-1. The addition of a cytokinin (BAP) in the medium led to lower embryo production, and this negative effect increased when the hormone concentration in the medium increased. The use of liquid medium and a dark incubation period immediately after the high temperature treatment were favourable for embryogenesis.     

Serial Subcultivation of Giardia lamblia in Keister's Modified TYI-S-33 Medium Containing Ultroser G

ABSTRACT. The ability of Giardia strains of the duodenalis type to grow in Keister's modified TYI-S-33 medium varies with serum lot. Recently, strains of Giardia including MR4, WB, and Human-1-Portland, have been cultivated in Keister's modified TYI-S-33 medium containing the serum substitute Ultroser G and have been cultured serially at least 40 times. An optimal concentration of 8% Ultroser G promotes maximal growth in Keister's modified TYI-S-33 medium for all three strains. This concentration of Ultroser G will produce a two-log increase in the number of trophozoites in approximately three days post-inoculation. Generation times for the trophozoites ranging from 6 to 11 h have been achieved in Keister's modified TYI-S-33 containing 10% adult bovine serum and from 8 to 13 h in Keister's modified TYI-S-33 with 8% Ultroser G. Despite the excellent growth of Giardia strains in medium containing Ultroser G, the maximum trophozoite density is only about half of that achieved in Keister's modified TYI-S-33 medium supplemented with 10% adult bovine serum. Comparisons of trophozoites grown with serum or the serum substitute reveal no discernable differences in morphology and motility. Additionally, these strains have been successfully cryopreserved and revived in Keister's modified TYI-S-33 medium supplemented with Ultroser G. Because Ultroser G is a characterized mixture of six main groups of ingredients (growth factors, adhesion factors, mineral trace elements, hormones, binding proteins, and vitamins), the variability in cell proliferation that may occur when changing serum lots should be minimized when using this product.     

Pathogenic fungal dynamics in a fall population of the blackmargined aphid (Monellia caryella)

A fall population of the blackmargined aphid, Monellia caryella (Fitch) (Homoptera: Aphididae), was monitored daily for mortality caused by fungi and parasitoids. Three species of Entomophthorales fungi were found. Fungal-induced mortality was found to be correlated with minimum temperature. During the period when the minimum temperature was consistently above 8°C, fungal-induced mortality was positively correlated with the sum of hours of leaf-wetness on days 5, 6, and 7 before sampling. Parasitoid wasps killed fewer hosts than fungi and were not affected by environmental factors.

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Résumé La mortalité quotidienne provoquée par des parasitoïdes et des champignons sur une population automnale de M. caryella Fitch (Homopt. Aphididae) a été suivie à Byron (Géorgie, USA), avec enregistrement de la température, de la pluie et de l'humidité des feuilles. 3 espèces de champignons entomophthorales ont été trouvées: Neozygites sp., Entomophthora planchoniana et Erynia sp. 54% des pucerons morts étaient contaminés par Neozygites et 46% par E. planchoniana. Seuls 2 pucerons avaient été tués par Erynia sp. Les taux de mortalité quotidienne par champignons s'élevaient après les périodes pluvieuses et d'humidité des feuilles. La mortalité due au parasitisme était inférieure à la mortalité due aux champignons et ne semblait pas liée aux conditions écologiques. La discussion porte sur l'importance de la mortalité provoquée par les champignons sur la dynamique de la population de pucerons.

     

Dibutyrylic cyclic AMP and theophylline inhibit proliferation of accessory cells in primary cultures of adrenomedullary cells

Summary Studies on isolated adrenal chromaffin cells in primary cultures may be seriously hampered by the presence of non-chromaffin, mainly fibroblast-like cells, which always occur in dissociates of adrenal medullary tissue and often outnumber the chromaffin cells by the end of the first week of culture, when no measures are taken to control their proliferation. The present study offers a new means to inhibit effectively the proliferation of these accessory cells by treating the cultures with dibutyrylic cyclic AMP (dbcAMP, 0.1 or 0.01 mM) and equimolar amounts of the phosphodiesterase inhibitor theophylline. With this treatment cultures of young rat adrenal chromaffin cells remain virtually free of accessory cells for two weeks of culture. Cultures of bovine adrenomedullary cells retain their initial amounts of non-chromaffin cells, which largely depends upon whether the primary cell suspensions have undergone differential plating prior to seeding. Suppression of accessory cell proliferation with dbcAMP and theophylline is partly due to maintaining differentiation of cortical cells, which otherwise dedifferentiate into rapidly dividing fibroblast-like elements. However, a more direct action of dbcAMP on accessory cells in terms of growth control is also conceivable. DbcAMP and theophylline in the doses applied do not impair the viability, ultrastructure and catecholamine-storing capacity of cultured chromaffin cells.     

Toxoplasma gondii: Purine synthesis and salvage in mutant host cells and parasites

Toxoplasma gondii, growing exponentially in heavily infected mutant Chinese hamster ovary cells that had a defined defect in purine biosynthesis, did not incorporate [U-¹⁴C]glucose or [¹⁴C]formate into the guanine or adenine of nucleic acids. Intracellular parasites therefore must be incapable of synthesizing purines and depend on their host cells for them. Extracellular parasites, which are capable of limited DNA and RNA synthesis, efficiently incorporated adenosine nucleotides, adenosine, inosine, and hypoxanthine into their nucleic acids; adenosine 5′-monophosphate was the best utilized precursor. Extracellular parasites incubated with ATP labeled with ³H in the purine base and ³²P in the α-phosphate incorporated the purine ring 50-fold more efficiently than they did the α-phosphate. Thus, ATP is largely degraded to adenosine before it can be used by T. gondii for nucleic acid synthesis. Two pathways for the conversion of adenosine to nucleotides appear to exist, one involving adenosine kinase, the other hypoxanthine—guanine phosphoribosyl transferase. In adenosine kinase-less mutant parasites, the efficiency of incorporation of ATP or adenosine was reduced by 75%, which indicates the adenosine kinase pathway was predominant. Extracellular parasites incorporated ATP into both the adenine and the guanine of their nucleic acids, so ATP from the host cell could supply the entire purine requirement of T. gondii. However, ATP generated by oxidative phosphorylation in the host cell is not essential for parasites because they grew normally in a cell mutant that was deficient in aerobic respiration and almost completely dependent upon glycolysis.