Polygalacturonase isoenzymes and oxalic acid produced by Sclerotinia sclerotiorum in soybean hypocotyls as elicitors of glyceollin

The polygalacturonases (PG) and oxalic acid produced by Sclerotinia sclerotiorum in infected soybean hypocotyls were investigated as elicitors of the phytoalexin glyceollin I.Purification to homogeneity through isoelectrofocusing and ion-exchange fast protein liquid chromatography revealed three endo-PG isoenzymes (PG-I, PG-II and PG-IV) and one exo-PG (PG-III) in 6-day-old etiolated soybean hypocotyls infected with the B-24 isolate of S. sclerotiorum.PG-I and PG-III, in the range of concentrations tested (0·15–1·2 reducing units ml⁻¹), did not act as elicitors of glyceollin I synthesis. Some elicitor activity was shown by PG-II at 0·6–1·2 reducing units ml⁻¹. PG-IV, at lower doses (0·038–0·30 reducing units ml⁻¹), was even more effective in inducing phytoalexin synthesis. However higher concentrations of PG-IV induced tissue softening and decreased phytoalexin accumulation.PG-II and PG-IV released heat-stable elicitors from purified soybean cell walls supporting the evidence that uronides are intermediate inducers in elicitation by endo-PGs. Oxalic acid was an active elicitor of glyceollin I over the range of concentrations tested (0·31–20 m ) with the maximum at a concentration of 5 m . The inability of oxalic acid to release uronides from purified cell walls makes it unlikely that uronide intermediate elicitors are involved in elicitation by oxalic acid.     

Tolerance ofsclerotinia sclerotiorum to glyceollin i,a soybean phytoalexin

Glyceollin I was fungistatic rather than fungicidal towardSclerotinia sclerotiorum. Within the mycelial mat apical cells were more vulnerable than mature cells.S. sclerotiorum removed large amounts of glyceollin from solution by a non-energy-requiring process. Contribution from Ministero dell’Università e della Ricerca Scientifica e Tecnologica.     

Study on the Acute Toxicity of Rare Earth Yttrium to Earthworms under the Stress of Leaching Agent Ammonium Sulfate

[目的]考察浸矿剂硫酸铵（NH4）2SO4胁迫下稀土离子钇的毒性研究。[方法]选择蚯蚓作为土壤生态环境污染指示生物,采用滤纸接触法研究硫酸铵胁迫下稀土钇（ Y）对蚯蚓的急性毒性效应。[结果]①稀土钇单一染毒,蚯蚓的48 h 半致死率浓度为 LC50=213.41 mg/L、24 h半致死率浓度为 LC50=322.63 mg/L。②硫酸铵单一染毒下,蚯蚓的48 h半致死率浓度为 LC50=13.89 g/L、24 h半致死率浓度为LC50=15.05 g/L。③低浓度10 g/L的硫酸铵与稀土钇复合染毒,蚯蚓的48 h半致死率浓度LC50=198.65 g/L、24 h 半致死率浓度 LC50=399.85 g/L;中浓度14 g/L的硫酸铵与稀土钇复合染毒,蚯蚓的48 h半致死率浓度 LC50=167.3 mg/L、24 h半致死率浓度 LC50=256.73 mg/L;高浓度20 g/L的硫酸铵与稀土钇复合染毒,蚯蚓的48 h半致死率浓度 LC50=31.03 mg/L、24 h的LC50=127.65 mg/L。[结论]低浓度的硫酸铵降低稀土钇对蚯蚓的毒性,产生一定的拮抗作用。中浓度的硫酸铵增加稀土钇对蚯蚓的毒性,产生较明显的协同作用。高浓度硫酸铵显著增加了稀土钇对蚯蚓的毒性。稀土钇染毒下蚯蚓死体更易断裂,而活体对针刺反应相对不灵敏。     

Embryonic and fetal development in - pigmy rice rat - Oligoryzomys sp. (Rodentia, Sigmodontinae) and its significance for being a new experimental model

Oligoryzomys (Cricetidae, Sigmodontinae) is a common rodent genus from South America that includes a couple of very similar species. Related species have been used as experimental model for understanding several diseases for which these species are reservoirs. In order to provide a better understanding of the embryological aspects of this group, herein we showed data on the embryonic and fetal development in Oligoryzomys sp. Eight specimens of different stages of gestation were obtained from the Collection of the Zoology Museum of University of Sao Paulo, Brazil. Gestational ages were estimated by crown-rump-length according to Evans and Sack (1973). To address our analysis after examining the gross morphology, tissues from several organs were processed for light and scanning electron microscopy. Morphological data on the systems (nervous system, cardiorespiratory system, intestinal tract and urogenital system) were described in detail. Finally, the findings were compared with what is known about embryological aspects in other rodent species in order to establish similarities and differences during the organogenesis in different species.     

Characterization of the extracellular matrix in the chorioallantoic membrane of water buffalo (Bubalus bubalis) in early gestation

Placenta is formed by a parenchyma rich in extracellular matrix (ECM), and this structure guarantees the proper development of the embryo and placental functioning. Recently, studies have focused on the characterization of ECM in the placenta and foetal membranes of different species. This work aimed to analyse the composition of the ECM and to quantify the types of collagens in its composition. For this, 33 chorioallantoic membranes were used at different gestational ages, which were grouped into five groups. Subsequently, haematoxylin–eosin staining, Masson trichrome and picrosirius were performed for histological analysis. Through the technique of polarized light, it was possible to quantify the total collagen present in the membranes and finally the immunohistochemical technique was performed to verify the presence of collagens and glycoproteins. It was possible to verify that the chorioallantoic membranes have, in all the gestational periods of the initial third of gestation, the same histological structures, being the most significant difference the membrane thickening that occurs gradually during the gestation. However, we notice the appearance of binucleate cells only from group II. In addition, it was verified that a gradual increase of collagen occurs until the group IV, yet from the group V begins to occur a decrease of this protein. In addition, collagen I, collagen III, fibronectin and laminin were present in all membranes. With this, we concluded that the buffalo chorioallantoic membrane presents ECM in constant remodelling at the beginning of gestation and can be used as biomaterial in works on regenerative biology.     

Polygalacturonase inhibiting proteins fromAllium porrumL. and their role in plant tissue against fungal endo-polygalacturonases

Five endo-polygalacturonases (PGs), three produced in culture filtrate byFusarium moniliforme, Sclerotium cepivorumandBotrytis aclada,respectively, and two (one acidic and one basic isoform) obtained fromSclerotinia sclerotiorumsoybean infected hypocotyls, were purified in order to characterize the activity of polygalacturonase inhibitor(s) (PGIP(s)) from leek stalk tissue (Allium porrumL.). Three apparently different PGIPs (PGIP-I, PGIP-II and PGIP-III) were purified from the leek tissue. The two more abundant PGIPs (PGIP-I and PGIP-III), although possessing similar pIs of about 6.5, differed in chromatographic behaviour, their molecular mass (39 and 42 kDa, respectively), and specific activity when assayed with the fungal endo-PGs. In addition, PGIP-I was solubilized from tissue homogenate with a low-salt buffer whilst PGIP-III needed a high-salt buffer for extraction (behaving as an ionically wall-bound protein). PGIP-II had very similar properties to PGIP-I, but was extracted using the high-salt buffer. The purified PGIPs and the crude leek extract showed similar inhibition activity patterns against the five fungal endo-PGs. The maximum inhibition activity was observed against the basic endo-PG fromS. sclerotiorum,followed by the acidic endo-PG ofS. sclerotiorumand the endo-PG fromB. aclada.In contrast, no inhibition of endo-PGs fromS. cepivorumandF. moniliformewas observed. Four different concentrations of the five fungal endo-PGs were incubated separately with slices of leek stalk, and the galacturonides released in the incubation mixture were measured. At every level used the endo-PGs ofF. moniliformeandS. cepivorumshowed the maximum activity in uronide releasing. The endo-PGs ofS. sclerotiorum(acidic PG) andB. acladawere active only when high levels were used while the basic endo-PG ofS. sclerotiorumwas not active in combustion with any level of PGIP. These results indicate that a close relationship exists between PGIP activityin vitroand the ability of PGIP to protect leek tissue from endo-PG degradation.     

Gross and microscopic anatomy of the pineal gland in Nasua nasua--coati (Linnaeus, 1766)

Nasua nasua, coati, is a mammal of the Carnivora order and Procyonidae family. It lives in bands composed of females and young males. The pineal gland or epiphysis of brain is endocrine, producing the melatonin. Its function is the control of the cycle of light environment, characteristic of day and night. For this research, five adult coatis were used, originating from CECRIMPAS-UNIfeob (Proc. IBAMA 02027.003731/04-76), Brazil. The animals were killed and perfusion-fixed in 10% formaldehyde. Pineals were measured and a medium size was found to be 2.3-mm-long and 1.3-mm-wide. Pineal gland was located in the habenular commissure in the most caudal portion of the third ventricular roof, lying in a dorso-caudal position from the base to the apex. Pinealocytes were predominantly found in the glandular parenchyma. Distinct and heterogeneous arrangements of these cells throughout the three pineal portions were observed as follows: linear cords at the apex, circular cords at the base of the gland, whereas at the body a transition arrangement was found. Calcareous concretions could be observed in the apex. The pineal gland was classified as subcallosal type [Rec. Méd. Vét.1, 36 (1956)] and as AB type [Prog. Brain Res. 42, 25 (1979); The Pineal Organ, Berlin/Heidelberg: Springer-Verlag (1981)].     

Fertility Results of Artificial Inseminations Performed with Liquid Boar Semen Stored in X-CellTM vs BTS Extender

The objective of the present field study was to compare the fertility results for boar semen diluted in X-cell stored up to 4-5 days before artificial insemination (AI) with semen diluted in Beltsville thawing solution (BTS) used for AI following 2-3 days of storage (where the first day being the collection day). A total number of 2601 double inseminations in Norwegian herds were included in this two-trial study. All the boars used in the study were mature cross-bred Norwegian Landrace x Duroc (LD), which were routinely used for AI in Norway. The inseminated gilts and sows were Norwegian Landrace x Yorkshire (LY). The AI doses contained 2.5 billion spermatozoa, and consisted of a mixture of semen from three, occasionally four, boars (i.e. heterospermic semen). Fertility was measured in terms of the likelihood of farrowing and subsequent litter size. The fertility of the semen in both of the extenders was satisfactory and no significant differences were found either in semen stored 4-5 days in X-cell compared with 2-3 days in BTS or in semen stored 2-3 days in X-cell compared with 2-3 days in BTS. The storage capability findings for the long-term extender X-cell could significantly simplify the practical issues of semen production and the distribution of AI doses containing 2.5 billion spermatozoa. However, in pig production systems where all semen is used within 2-3 days, the short-term extender BTS is as good as the more expensive extender X-cell.