ULTRASTRUCTURAL ASPECTS OF SHOOT INITIATION IN TOBACCO CALLUS CULTURES

An ultrastructural investigation of shoot initiation in tobacco (Nicotiana tabacum L. var. W. 38) callus cultures was made. Zones of preferential division were observed in the basal portion of the tissue by eight days in culture and these led, sequentially, to meristemoids, primordia, and shoots. During the initial stages of meristemoid formation, protein inclusions and large accumulations of plastid starch were present in the cells, while vacuoles were filled with membranous and cytoplasmic protrusions. At later stages of meristemoid development, these features were not observed in the cells, which were also smaller in size and possessed numerous small, peripheral vacuoles. It appears that the membranous and cytoplasmic protrusions are involved in vacuolar reduction during meristemoid formation. It would also appear that the storage materials supply the energy and other reserves needed for the organogenetic process. By contrast, tissue cultured under nonshoot-forming conditions and nonmeristemoid regions of shoot-forming tissue remained parenchymatous over the same time period.     

疫霉属菌种的液氮超低温保藏

对疫霉属的15个种68株菌和霜疫霉2株菌进行了液氮超低温的保藏试验并得到成功。比较了冷冻速度、保护剂和解冻速度对菌存活的影响。严格控制每分钟降温摄氏1度直到-40℃后再放入液氮罐中,对疫霉和霜疫霉来说都是非常必要的。这种降温程序可通过简单设备人工操作达到。而直接由室温降到-150℃以下会损伤菌种以致死亡。在所用的保护剂中,不论10％甘油还是5-15％二甲基亚砜都能起到保护作用。尚看不出对那种保护剂有特别的要求,似可任意选用。至于解冻条件,由液氮中取出放置在38℃水浴中快速融化与在20℃水浴中中等速度融化效果相等,对菌的成活都没有太大影响。我们的试验肯定了在有保护剂存在下,用慢速冷冻可以在液氮中保存疫霉和霜疫霉。     

MORPHOGENETIC ASPECTS OF MULTILAYERING IN PETRI DISH CULTURES OF HUMAN FETAL LUNG FIBROBLASTS

Randomly seeded Petri dish cultures of embryonic human lung fibroblasts generate, in the course of their growth, highly ordered cellular arrangements. Thick, bilaterally symmetrical ridges with an axial polarity and an orthogonal, multilayered internal organization are observed within stationary cultures. The generation of these structures has been investigated. Ridges result from the spontaneous aggregation of cells in postconfluent cultures brought about by directed cell movements. These movements are promoted by the localized production of extracellular matrix sheets containing collagen, which provide new substrates for cellular colonization. Cells that have colonized one matrix substrate may secrete another above themselves, which will in turn be colonized. By a continuation of this cycle, thick stacks consisting of alternate layers of cells and matrix are produced to yield the observed aggregations. The distribution and shape of ridges in a culture imply that matrix substrates are confined to specific locations. The suggested control hypothesis assumes that all the cells in fibroblast cultures are potential producers of a single species of matrix. The serviceability of this matrix as a substrate for cellular colonization, however, is destroyed if the producer cells are motile. Matrix substrates, therefore, are only made by nonmotile cells.     

SOME ASPECTS OF THE PHYSIOLOGY AND BIOCHEMISTRY OF MARINE FUNGI

1. This review deals with certain aspects of the physiology and biochemistry of marine fungi. Though it is not easy to define what is meant by a marine fungus, the information presented here relates to those species that by consensus can be accepted as being truly marine. The material is in two parts, that relating to the higher fungi (Ascomycotina, Basidiomycotina and Deuteromycotina) and that relating to the lower fungi, particularly those zoosporic fungi that require sodium for growth. 2. Higher marine fungi appear to have a similar carbon, nitrogen and vitamin nutrition to their terrestrial counterparts. 3. Growth of higher marine fungi can be optimal in 100% sea water, but more frequently optimal growth is at a lower percentage. The percentage giving optimal growth may be determined by the rate of ion uptake required to generate the necessary turgor for growth. The tolerance of the vegetative phase of many terrestrial fungi to salinity appears little different from that of marine fungi. In general, members of the Basidiomycotina are particularly sensitive to salinity, while those of the Ascomycotina and Deuteromycotina are much more tolerant. 4. The degree of tolerance to salinity may be dependent upon temperature and whether or not adaptation by the fungus has occurred. 5. Maintenance of a suitable internal potassium concentration by a higher marine fungus is important for growth in a saline medium. Calcium appears to be necessary for the retention of potassium and organic solutes within the hyphae. 6. It appears that higher marine fungi are able to maintain a ratio of potassium: sodium higher than that of sea water by the presence of a plasma membrane ATPase which may have a higher pH optimum than the equivalent enzyme from terrestrial counterparts. 7. Higher fungi produce more glycerol as the salinity of the external medium is increased. Whether or not the compound is involved in osmoregulation has yet to be determined. 8. Turgor is thought to be generated in higher marine fungi growing in sea water by organic solutes (predominantly glycerol and arabitol) and by ions, with the latter playing the major role. Though interpretation of the data depends on several assumptions, the high concentrations of sodium that seem likely to be present in hyphae or cells have implications for the activity of enzymes, if they have similar properties to those of higher plants. There is a need for information on the effects of high concentrations of ions on enzymes located in the cytoplasm of higher fungi. 9. In spite of some experimental uncertainties, it seems that reproduction and spore germination of higher marine fungi are very much less affected by salinity than are the same processes of terrestrial counterparts. 10. The zoosporic marine fungi require sodium for growth. Though the ion is required at high concentration for growth, sodium cannot be replaced by potassium. Evidence indicates that sodium is involved in the transport of solutes across the plasma membrane. 11. The carbon and nitrogen requirements for the growth of the zoosporic marine fungi demand further investigation, particularly at the biochemical level. There is evidence that the respiration of these fungi possesses many interesting features. 12. Vitamin requirements of the zoosporic marine fungi depend on the isolate under investigation. Vitamin B., does not now seem to be an obligatory requirement. The ability of phospholipids and sterols to stimulate growth requires further investigation. 13. Further studies on marine fungi in the laboratory should focus particularly on growth in continuous culture. Physiological and biochemical studies would be helped by more precise guidance from those concerned with the ecology of these fungi.     

液氮保存曲霉菌种的效果

本文报告了以蒸馏水、10%甘油和5%二甲基亚砜(DMSO)为保护剂,用液氮冻结法保存5种8株曲霉的效果,并检测了这些菌分别产生的亚甲基丁二酸、柠檬酸、蛋白酶和糖化酶的生理活性。这些菌在液氮气相(接近-150℃)中保存180天全部保持着生活能力,它们的培养特征和形态特征保留原来的形状。所测定的液氮保存8株菌种的生理活性,除两株糖化酶活力稍有降低外,其它菌株没有明显的变化。     

A SHAKING DEVICE FOR LIQUID CULTURES

SUMMARY: Details are given for the construction of a reciprocating shaking device for liquid cultures. Up to 36 litres can be shaken at one time. The machine is compact, the base being only 28 in. square.     

THE ABSORPTION OF OXYGEN FROM LIQUID CULTURES OF BLUE-GREEN ALGAE BY ALKALINE PYROGALLOL

SUMMARY

A mixture of pyrogallol with sodium hydroxide and sodium carbonate (2:1:1) in a separate container absorbed up to 75% of the oxygen in a bicarbonate-carbonate buffered medium with more or less no change in the pH of the medium. A system in which two test tubes were added to the culture vessel, one containing saturated sodium bicarbonate and the other pyrogallol plus sodium hydroxide gave similar results, but were not investigated further because of difficulties in handling. Optimal conditions for absorption of oxygen from 100 ml of the growth medium was 1 mMole pyrogallol +0,5 mMole sodium hydroxide + 0,5 mMole sodium carbonate. The addition of this pyrogallol to alkali ratio to cultures of two Microcystis and one Synechococcus isolate in rubber stoppered flasks gave stoichiometric increases in yield which was not due to carbonate enrichment but to a lowering of oxygen tension. The data may mean that even under relatively low light intensities (1 - 4 × 10³ ergs cm^?2 sec^?1) photooxidation occurs.     

细胞培养物中的逆转录病毒检测

本文用非放射性法测定了K562,SP2/0,L-02,SPC-A-1,SMMC-7721和Hut-102等6个细胞培养液超离心沉淀物逆转录酶的含量,间接证明细胞培养物中逆转录病毒的存在。结果表明,Hut-102细胞为阳性,其它5个细胞系为阴性。     

THE CONTROL OF SEX IN FUNGI

Raper , John R. (Harvard University, Cambridge, Mass.) The control of sex in fungi . Amer. Jour. Bot. 47(9) : 794–808. 1960.—The control of sexual processes in fungi is imposed at many successive points in the developmental cycle. Two major categories of controlling factors are recognized: (1) a predetermination of future sexual capacity follows from the segregation at meiosis of genetic factors that impose differentiation in respect to sexual sign or incompatibility type; (2) a coordinated progression of interdependent stages, governed by numerous genetic-metabolic factors, leads to plasmogamy, karyogamy, and meiosis. The major features of these 2 types of control—the several patterns of sexuality of the former and sexual hormones and sequential sexual development of the latter—are reviewed, and the possible relationship between the two levels of control is considered in some detail.