青霉素和NAA与6—BA配合在离体草莓器官建成中的作用

本实验研究了青霉素和ＮＡＡ与６－ＢＡ配合对离体草莓器官建成以及体内过氧化物同工酶的影响。结果表明：青霉素促进不定根的分化,但抑制不定根的伸长和不定芽的分化,并且不同程度地影响器官建成过程中过氧化和抽工酶的谱带和活性。青霉素与植物激素一样,参与了植物体内的生理代谢而引起植物器官建成。     

Continuous measurement of changes in intracellular calcium concentration in mouse splenic T cells attached to a glass substrate

Mitogen- and isoproterenol-induced changes of [Ca²⁺]_i in T cells attached to a glass substrate were examined. Murine (C57BL/6) splenic T cells were attached to coverslips or 35-mm dishes (MatTek) precoated with Cell Tak® (3.5 µg/cm²). The cells were then loaded with fluorescent dye (2 µg/ml of fura2-AM or fluo3-AM) and changes in [Ca²⁺]_i in a population of cells (using a spectrofluorometer) or in single cells (using a confocal microscope) were measured during continuous superfusion. Population measurements of [Ca²⁺]_i demonstrated that concanavalin A (Con A, 2 or 5 µg/ml) caused an increase in [Ca²⁺]_i that rose to a peak and then declined to a steady state. The concentration-response relationship (0.05–5 µg/ml) had an EC₅₀ of 0.3 µg/ml. Isoproterenol decreased the Con A-induced elevation of steady state [Ca²⁺]_i. In single cell studies, the increase in [Ca²⁺]_i in response to Con A typically occurred in about 50% of the cells in a microscope field, and the delay before activation varied among cells. Taken together, these data demonstrate that Cell Tak® can be used to attach T cells to glass coverslips and will be useful for the study of signaling mechanisms in T cells.     

Determination of base specificity of multiple ribonucleases from crude samples

Ribonucleases are widely found in the tissues of living organisms, but the functions of individual ribonucleases are not clear. To facilitate characterization of individual ribonucleases, I have developed a rapid method to separate and identify each ribonuclease from a crude sample by gel electrophoresis instead of by time-consuming purification steps. The ribonucleases in a crude sample are first separated by RNA-cast SDS-polyacrylamide gel electrophoresis and then eluted from the gel after ethidium bromide staining. To determine the base specificity of each ribonuclease, a 5 labelled oligonucleotide with known sequence is added to the enzyme eluate and the digested products are analyzed by denaturing gel electrophoresis. The base specificity of bovine pancreatic ribonuclease (RNase A), bullfrog oocyte-specific ribonuclease (RC-RNase), human serum ribonucleases and sweet potato leaf ribonucleases were determined by this method. Other properties of individual ribonucleases, e.g. substrate preference, may also be determined from crude samples by this method without further purification steps.Abbreviations RNase ribonuclease - SDS sodium dodecyl sulfate     

The effect of ethanol on lateral and rotational mobility of plasma membrane vesicles isolated from cultured mar 18.5 hybridoma cells

Intramolecular excimer formation of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of the lateral mobility and the range of the rotational mobility of bulk bilayer structures of the plasma membrane vesicles (ATCC-PMV) isolated from cultured hybridoma cells (ATCC TIB 216). In a concentration-dependent manner, ethanol increased the excimer to monomer fluorescence intensity ratio (I/I) of Py-3-Py in the ATCC-PMV and decreased the anisotropy (r), limiting anisotropy (r) and order parameter (S) of DPH in the ATCC-PMV. This indicates that ethanol increased both the lateral and rotational mobility of the probes in the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was utilized to examine the range of transbilayer asymmetric rotational diffusion of the ATCC-PMV. The anisotropy (r), limiting anisotropy (r ) and order parameter (S) of DPH in the inner monolayer were 0.024, 0.032, and 0.069, respectively, greater than calculated for the outer monolayer of the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was also used to examine the transbilayer asymmetric effects of ethanol on the range of the rotational mobility of the ATCC-PMV. Ethanol had a greater increasing effect on the range of the rotational mobility of the outer monolayer as compared to the inner monolayer of the ATCC-PMV. It has been proven that ethanol exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the ATCC-PMV.This paper was supported in part by a research grant from the Korea Science and Engineering Foundation (KOSEF 88-1013-01) and from the Korea Research Foundation (1991–1993).     

Expression and secretion of human lipocortin-1 by promoter and signal sequence of STA1 from Saccharomyces diastaticus

Summary For the secretion of human lipocortin-1 (LC-1) in yeast, a expression and secretion vector was constructed by using the promoter and signal sequence of glucoamylase gene (STA1) of Saccharomyces diastaticus. After the cDNA of human LC-1 was ligated with the secretion vector, the resulting hybrid plasmid was transformed into S. diastaticus. When the recombinant S. diastaticus was cultivated in YPD medium, LC-1 was expressed and secreted into the extracellular medium, yielding LC-1 protein at a concentration of 2.5 g/mL.     

稻田土壤－作物－家畜系统中氮的循环研究

本文用Ｎ￣（１５）标记水稻和绿肥研究了稻田土壤－作物－家畜系统中氮的循环。Ｎ￣（１５）标记稻草喂羊,羊体回收饲料稻草Ｎ３１．１６％,羊粪２８．２６％,羊尿５．７２％,总回收６５．１４％,损失３４．８６％。将羊粪尿单施,稻谷回收饲料稻草Ｎ３．１９％,水稻全株回收４．８２％,土壤残留１９．００％,损失１０．１４％。故羊体、水稻及土壤残留共回收饲料稻草Ｎ５４．９８％．将羊粪与尿素配施,则饲料稻草Ｎ的总回收率为５５．８８％。Ｎ￣（１５）标记绿肥喂猪;猪体回收饲料绿肥Ｎ２３．５１％．猪粪２３．８５％,猪尿２８．７６％,总回收率７６．１２％,损失率２３．８８％。将猪粪、尿还田,稻谷回收饲料绿肥Ｎ６．６９％,水稻全株回收１０．０５％,土壤残留１９．１７％,故猪体、水稻和土壤残留共回收饲料绿肥Ｎ５２．７３％,将猪粪与尿素配施,则饲料绿肥Ｎ的总回收率为５２．７５％。     

广东种子植物区系地理成分研究

广东种子植物区系共有２１９科１４３４属４９８６种。其中：①热带区系成分占有较大的比例,约占非世界属的６２．３％,表明广东区系受到热带区系的强烈渗透,次之为亚热带（至亚热带山地）成分约占３２．２％,及中国特有分布占５．６％;②以单型属（７２属）,单种属（５８５属）和寡种属（５６２属）占绝对优势,共占８５．０％,充分显示了广东区系的古老性和过渡性特点;③从区系组成和植被结构的特征成分来看,以华夏植物区系成分为主,包括：亚热带分布,亚热带山地分布,中国特有分布,东亚—北美间断分布,及一些亚洲热带分布中以亚热带为主的属：④在这些属中有东亚特有属１３２属,中国特有属７５属以及华南—西南—中南半岛特有属６８属等;这些属的存在毫无疑问地说明,广东区系是华夏植物区系的核心地区之一,同时也显示了热带区系与亚热带区系有着不可分隔的统一性。     

Regeneration of haploid plants from isolated microspores of asparagus (Asparagus officinalis L.)

High percentages of micro-calli and micro-derived embryos were produced from isolated asparagus microspores at late uninucleate stage on MS liquid medium supplemented with 1.0 mg l^–1 2,4-D and 0.5 mg l^–1 BA. Two types of calli, namely compact callus (CC) and loose callus (LC), were found. Plantlets were regenerated via organogenesis, when these calli were transferred onto MS solid medium supplemented with 1.0 mg l^–1 BA and 0.2 mg l^–1 IBA 6 weeks. Embryos were produced from liquid cultured microspores, or from solid cultured micro-calli. The frequencies of haploid plant production from organogenesis and embryogenesis were compared. Effects of plant growth regulators on callus production, plantlet regeneration, and haploid plant production were tested. The combination of BA 1.0 mg l^–1 and IBA 0.2 mg l^–1 resulted the highest precentage of haploid plant production (7.7% from CC, 4.3% from LC).Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IBA 3-indolybutyric acid - BA 6-binzyladinine - NAA naphtalene acetic acid - MS Murashige and Skoog