Preliminary Studies on the Photosynthetic Structures of Trifolium alpinum L. as Related to Productivity

Trifolium alpinum L. is a high-quality alpine forage plant growingspontaneously from 1900 to 2800 m above sea level and is widelydistributed in Piedmont and the Valle d'Aosta (Italy), whereit can reach population frequencies of 90 per cent. Yields weredetermined on forage harvested in the Valle dell'Orco (Piedmont)and were comparable to cultivated clovers from higher latitudes;yields decreased progressively as the elevation increased. Thechemical and nutritional characteristics of the forage, thoughcomparable to clovers cultivated in the Po valley (Italy), were,however, more constant. The structure of the leaf lamina asrelated to elevation was investigated using light microscopy,TEM and SEM. This is complemented by data on chlorophyll concentration,succulence, specific leaf weight and area. At all elevationsT. alpinum lacks, apart from bundle sheath cell chloroplastsin a centrifugal arrangement, the structural characteristicsof C₄ plants. The chlorophyll a:b ratio (less than four) istypical of a C₂ plant. Succulence indices (S and S_m) were verylow, making CAM pathway photosynthesis unlikely. Unusual anddifficult to interpret structures included: small functionalchloroplasts in both the epidermises, stomata present almostexclusively in the upper epidermis and mitochondria enveloped(or enclosed) by chloroplasts. It was observed that, as theelevation increases, populations are selected which are well-adaptedfor gas exchange (increase in specific leaf area, stomatal densityand intercellular spaces) and characterized by a decrease inthe grana thylacoid:integrana thylacoid ratio (consistent withthe increase in the chlorophyll a:b ratio), the per cent water,S_m and the specific leaf weight. Trifolium alpinum L., alpine trefoil, leaf structures, photosynthesis, yield, elevation, C₂, C₄     

PCR detection of MLOs in quick decline-affected pear trees in Italy

Polymerase chain reaction (PCR) amplification, using primers derived from the 16S rRNA gene, followed by restriction fragment length polymorphism (RFLP) analysis with Alu I restriction endonuclease was used to detect myc-oplasma-like organisms (MLOs) associated with pear decline. MLOs were consistently detected in pear trees that suddenly wilted and died within a few days during summer, as well as in pears of the same orchards with symptoms similar to the slow form of pear decline. In both cases the same RFLP pattern was obtained. Declining pear trees were 5 to 8-yr-old cvs Williams, Kaiser and Max Red Bartlett grafted on to Pyrus communis seedling rootstocks. All the orchards affected by quick decline had severe attacks of pear psyllid (Cacopsylla pyri) during the year this study was performed and during the previous year. The results showed the suitability of DNA amplification by the polymerase chain reaction for the detection of pear decline MLOs and established that MLOs can be detected in infected tissues of dead trees.     

Chemical and Electron Microscopic Studies of Cattle (Bos taurus) With Four Types of Phenotypic Pigmentation

The biological behavior of the pigmentary phenotypes of four breeds of cattle has been analysed: the black pigmentation of Holstein Friesian; the red pigmentation of Limousin; the dilution in Charolais; and the postnatal disappearance of red pigmentation in Chianina. The analytic techniques included the characterization of melanins by high-performance liquid chromatography, the examination of follicular melanocytes by light microscopy, and the examination of melanosomes by electron microscopy. The black phenotype was very strongly eumelanogenic. The red phenotype in Limousin is polymorphic: individual follicular melanocytes contain both mature eumelanosomes and pheomelanosomes. Charolais and Chianina cattle exhibited a dramatic reduction in melanogenic activity, which was characterized by the almost exclusive presence of prephaoemelanosomes in Charolais and of immature premelanosomes in Chianina. In the dilute Charolais phenotype, the density of distribution of follicular melanocytes also seemed to be reduced. The genes that are responsible for these four phenotypes seem to act on the maturation, differentiation, and density of distribution of the melanosomes.     

PRIMORDIAL GERM CELL-DERIVED MOUSE EMBRYONIC GERM (EG) CELLSIN VITRORESEMBLE UNDIFFERENTIATED STEM CELLS WITH RESPECT TO DIFFERENTIATION CAPACITY AND CELL CYCLE DISTRIBUTION

Embryonic germ (EG) cells of line EG-1 derived from mouse primordial germ cells were investigated for theirin vitrodifferentiation capacity. By cultivation as embryo-like aggregates EG-1 cells differentiated into cardiac, skeletal muscle and neuronal cells accompanied by the expression of tissue-specific genes and proteins as shown by RT-PCR analysis and indirect immunofluorescence. In comparison to embryonic stem (ES) cells of line D3 the efficiency of differentiation into cardiac and muscle cells was comparatively low, whereas spontaneous neuronal differentiation was more efficient than in D3 cells. Furthermore, the distribution of cell cycle phases as a parameter for the differentiation state was analysed in undifferentiated EG cells and ES cells and compared to data obtained for embryonic carcinoma (EC) cells of line P19 and differentiated, epithelioid EPI-7 cells. Flow cytometric analysis revealed similar cell cycle phase distributions in EG, EC and ES cells. In contrast, the somatic differentiated EPI-7 cells showed a longer G₁-phase and shorter S- and G₂/M-phases. Together, our results demonstrate that the differentiation state and capacity of EG cellsin vitroresemble that of totipotent ES cells.     

Studies on Spermatogenesis and Steroidogenesis in Culture

This paper summarizes the major pertinent findings of previouslypublished work and includes new experimental data. Culture conditionshave been defined for long-term maintenance of basic testicularstructure, primitive type A spermatogonia, and Sertoli cells.The primitive type A spermatogonia grown for 8 weeks in organculture were capable of restoring complete spermatogenesis afterthe cultures were transplanted into the testes of sexually maturehomologous hosts. Since the primitive type A spermatogonia werethe only germinal elements present in the organ cultures atthe time of transplantation, they probably represent the spermatogenicstem cells. Differentiation of gonocytes or spermatogonia tospermatocytes in late pachytene stage required presence of vitaminsA, C, and E or 4 mM concentration of glutamine in the mediumbut was not affected by hormones. The replication of Sertolicells in vivo and in organ culture was shown to be age relatedand hormone independent. The Sertoli cells possess specificFSH binding receptors and respond to FSH with increased synthesisof c-AMP and androgen-binding protein. The biologic functionof FSH in the Sertoli cells remains to be clarified. Other cellularcomponents of the testis, the germinal, or peritubular cellsdo not bind FSH. In organ cultures of testicular tissue or Leydig-cellcultures all enzymes in the steroid biosynthetic pathway, leadingfrom progesterone to testosterone, remain active for severaldays. In older cultures, the conversion of progesterone to testosteronebecomes impaired primarily due to loss of two specific enzymes:17 hydroxylase and 17-20 lyase. Addition of HCG to monolayercultures of Leydig cells stimulated both the synthesis and releaseof testosterone.     

The effect of salinity on larval survival and development in the sea urchin Echinometra lucunter

Summary

Reductions in salinity can have adverse effects on larval development and larval survival in some invertebrate taxa but not others. Salinity tolerance of larvae may be particularly important in echinoderms because they are both poor ion regulators and stenohaline. I examined the effect of six levels of salinity (15, 18, 21, 24, 27 and 33 PSU) on survival and rate of development of larvae in the subtropical sea urchin Echinometra lucunter. In the short-term, mortality rate was significantly lower in 33 PSU than in all other salinities except 27 PSU, and it was significantly greater in 15 and 18 PSU than in all higher salinities. In the long-term, daily and cumulative mortality were significantly greater in 15 PSU than in most other salinities over 11 days of development (except for cumulative mortality in 18 PSU). They were significantly greater in 18 PSU than in 21 PSU or 33 PSU over a period of 13 days. Furthermore, daily mortality was significantly greater in 18 PSU than in 24 PSU or 27 PSU at 13 d after fertilization. Daily and cumulative mortality were significantly lower in 33 PSU than in 21, 24 or 27 PSU over a period of 17 days. Although in the control (33 PSU) 75% of larvae completed development to the 8-arm stage at 35 d, no larvae developed further than the 4-arm stage in 18, 21, 24 or 27 PSU; in 15 PSU, ~60% of larvae did not develop further than swimming blastulae. Since prolonged exposure to salinities as high as 27 PSU (frequently recorded in the adult habitat) can result in great larval losses, adaptive behaviours that prevent larvae from entering water layers of low salinity will enhance their chance for survival.