CHANGES IN RESPONSE TO PURINE AND PYRIMIDINE DERIVATIVES OF CARROT ROOT CALLUS DURING SUCCESSIVE CULTURING

1. The growth of the carrot root callus which had been subculturedfor a long period (CCL) was promoted by the addition of 5l0^–8and 5l0^–7 M kinetin, whereas in the callus subculturedfor a short period (CCS) no growth promotion was observed atany concentrations of kinetin tested.
2. CCL showed an increasedgrowth in response to the applicationof kinetin, guanine, adenine,hypoxanthine, uracil, thymine,and cytosine in the presenceof fractions A and C of carrotroot extract, whereas no suchresponse was observed in CCS.CCL required fraction C to respondto uracil and probably purineand pyrimidine derivatives ingeneral.
3. The growth of CCL was promoted by kinetin, guanine,adenine,or hypoxanthine in the medium containing inositol andaminoacids mixture. In this case the growth-promoting actionof guanine,adenine, or hypoxanthine was nullified by kinetin.

(Received December 24, 1964; )     

CHANGES IN THE RESPONSE OF CARROT ROOT CALLUSES TO THE ROOT EXTRACT OCCURRING DURING THEIR SUCCESSIVE CULTURES

1. Alcohol extract of carrot root promoted the growth of the carrotroot callus which had been succesively cultured for more than18 months (CCL) on the medium containing WHITE'S inorganic salts,sucrose, yeast extract and 2, 4-D, but only a weak promotionwas observed for the growth of the carrot root callus whichhad been cultured for less than 14 months (CCS).
2. The activesubstances were fractionated by Amberlite IR-120and AmberliteIRA-400 into four fractions; C, D, E, and F. Eachfraction seemedto act synergistically to produce the effectof the whole carrotroot extract on the growth of CCL.
3. Fraction F of the carrotroot extract, which was adsorbed byAmberlite IRA-400 but notby Amberlite IR-120, promoted thegrowth of CCL in the presenceof other fractions, but had noeffect on the growth of CCS.So the different responses to thealcohol extract of the carrotroot calluses having differentlengths of successive cultureperiod seemed to depend mainlyon the ability of respondingto fraction F.
4. Using four strains of carrot root callusesof different origin,it was ascertained that different responsesof carrot root callusesto fraction F depended on the lengthof their culture and noton their strain-specific characters.
5. The substances active for the growth of CCL in the carrotrootextract passed through a dialysis membrane. These substanceswere little affected by autoclaving and remained in the aqueouslayer when shaken with several organic solvents: n-butanol,ethyl acetate, chloroform, benzene, ethyl ether and carbon tetrachloride.
6. Alcohol extract of carrot root also promoted the growth ofcarrotroot explant, tobacco stem callus and sunflower crowngall tissue.

(Received December 24, 1964; )     

胡萝卜愈伤组织形成过程中激素和创伤诱导的钙调素水平的变化

CaM抑制剂TFP抑制了胡萝卜愈伤组织的形成。ELISA法测定CaM的动态表明,愈伤组织形成过程中,出现了两个CaM 的峰。第一个峰值在6h,可能是由创伤引起的,单位蛋白和单位鲜重的CaM 量都增加了1倍左右,第二个峰值可能是由激素引起的,从48 h以后开始增加,7天时单位鲜重的CaM 增加了4倍左右,单位蛋白的CaM 量也特异地增加了0.7倍,此结果亦为CaM 免疫荧光组织化学定位结果所证实。研究结果表明胡萝卜愈伤组织形成过程中激素与创伤效应和CaM 动态有关。     

CHANGES IN ORGAN FORMING CAPACITY OF CARROT ROOT CALLUSES DURING SUBCULTURES

1. The formation of buds and roots in seven strains of carrotrootcallus successively cultured for various periods on a mediumcontaining WHITE'S inorganic salts, sucrose, 2,4-D and yeastextract was investigated. 2,4-D completely suppressed organformation during stock subculturing. It was confirmed that theorgan forming capacity of the callus in a 2, 4-D-free test mediumdiminishes and finally completely disappears with prolongedperiods of previous subculturing of the callus.
2. IAA promotedthe root formation. Yeast extract, casein hydrolysateand aminoacids mixture promoted the bud formation of callusesat earlieststeps of subculturing (Phase I).
3. At next steps of subculturing(Phase II), IAA-dependent rootforming capacity of calluseswas lost although the bud formingcapacity induced by yeastextract, casein hydrolysate and aminoacids mixture was retained.
4. At further advanced steps of subculturing (Phase III), yeastextract induced only root formation, while casein hydrolysatestill could induce buds. IAA and amino acids mixture did notaffect the organ formation.
5. No organ formation was observedin calluses subcultured over38 months under any conditionsattempted (Phase IV).
6. Single cells or small cell clumps obtainedfrom the callus subculturedfor 2 months formed only roots onthe medium containing IAAand formed buds and roots on the mediumcontaining yeast extract.
7. These differences in organ formingcapacity and in responsestowards various factors are interpretedto reflect the changesin physiological states of the callusduring successive cultureson the stock culture medium.

(Received December 24, 1964; )     

烟草愈伤组织分化和芽原基形成期间呼吸代谢途径的改变

接种在继代培养基上的柳叶烟草愈伤组织,未观察到组织分化和芽原基形成。在分化培养基上生长的愈伤组织,接种后第6天可见拟分生组织和管胞分化,9—12天有芽原基形成,15—18天可观察到苗端结构。根据碘乙酸、Na_3PO_4和丙二酸抑制试验,以及3-磷酸甘油醛脱氢酶与琥珀酸脱氢酶活性测定结果,初步表明烟草愈伤组织呼吸中存在有EMP、HMP和TCAC代谢途径.在发生输导组织和芽原基分化的愈伤组织中(接种后第6—12天),HMP途径的运行程度较高;而芽原基的继续生长(培养12天以后),则与EMP途径的增加有关;分化培养基上生长的愈伤组织,始终较继代培养愈伤组织具有较高的FCAC活性水平。     

LIGHT AND HORMONAL CONTROL OF ROOT FORMATION IN ZEA MAYS CALLUS CULTURES

Callus cultures of Zea mays were used to study the interaction of light with exogenous cytokinin/auxin levels in the initiation, growth and development of roots. Three auxins, indoleacetic acid (IAA), naphthaleneacetic acid (NAA) and 2,4 dichlorophenoxyacetic acid (2,4 D) were remarkably different in their effects on callus growth and root initiation. NAA at concentrations of 5 and 25 μM produced the highest combined yields of callus and roots under low light conditions. No significant morphological effects on roots were observed with the three auxins tested nor did low and intermediate light intensities alter root development.
At intermediate light levels the addition of the cytokinin, zeatin, was also able to influence the differentiation of the callus tissue. Increasing the cytokinin/auxin ratio from low to high shifted the development from callus growth to abundant root formation. High light caused the formation of short, thick roots. This effect could be counteracted in part by zeatin which promoted elongation. These observations suggest that both, the cytokinin/auxin ratio and light play an important role in the development of monocotyledonous roots.     

NUCLEOLAR AND BIOCHEMICAL CHANGES DURING UNBALANCED GROWTH OF TETRAHYMENA PYRIFORMIS

Numerous nucleoli can be observed in the macronucleus of the logarithmically growing ciliated protozoan Tetrahymena pyriformis; at late log phase the nucleoli aggregate and fuse. In stationary phase this fusion process continues, leaving a very few large vacuolated nuclear fusion bodies in the nucleus. When these stationary phase cells are placed into fresh enriched proteose peptone medium, the large fusion bodies begin to disaggregate during the 2.5-hour lag phase before cell division is initiated. By 3 to 6 hours after inoculation the appearance of the nucleoli in many cells returns to what it was in logarithmic cells. In view of the possible role of nucleoli in ribosome synthesis, attempts were made to correlate the morphological changes to changes in RNA and protein metabolism. The beginning of an increased RNA synthesis was concomitant with the beginning of disaggregation of the large fusion bodies into nucleoli, which was noticed in some cells by 1 hour after the return to fresh enriched proteose peptone medium. Increased protein synthesis then followed the increased RNA synthesis by 1 hour. The supply of RNA precursors (essential pyrimidines) were removed from cultures which were grown on a chemically defined synthetic medium, in order to study the relation between nucleolar fusion and synthesis of RNA and protein. Pyrimidine deprivation drastically curtailed RNA and protein synthesis, but did not cause fusion of nucleoli. When pyrimidines were added back to this culture medium, RNA synthesis was immediately stimulated and again preceded an increased protein synthesis by 1 hour. These studies suggest the involvement of unfused nucleoli in RNA and protein synthesis and demonstrate the extreme plasticity of nucleoli with respect to changes in their environment.     

胡萝卜光自养型愈伤组织的诱导培养及其光合特性

经诱导得到胡萝卜光自养型愈伤组织。以叶绿素为单位计算,获得的光自养型愈伤组织的光合活力达到甚至超过了整体植株叶片水平。同时测定愈伤组织光自养过程中光合特性的变化,结果表明其叶绿素含量逐渐上升、暗呼吸速率和Chl a/Chl b比值逐渐下降。并且用电子显微镜观察到愈伤组织中叶绿体结构逐渐发育的过程。     

黄瓜果实贮藏过程中某些形态,生化变化及其控制

黄瓜果实采后贮藏过程中,随着其花端部位的种子的不断发育,中部及茎端逐渐萎缩变糠,花端膨大(称为“大头现象”)。花端胎座组织中蛋白质、DNA、RNA 及干重等都随着时间延长呈增加趋势,而中部组织的上述物质的含量则急剧下降。贮藏20天,中部果皮及胎座组织中 DNase 活性分别增加了5和7倍,RNase 活性增加了5和6倍,这两种酶活性在花端组织中则几无变化。实验发现用 BA+GA,处理果实对于延迟花端膨大及中部萎缩有效,并能显著延缓果实叶绿素含量的降低。未授粉果实在贮藏过程中不出现“大头现象”,其蛋白质等物质的含量在果实各部位几乎以相同的速度降低。     

ATTEMPTS TO OBTAIN BACTERIA-FREE PLANTS OF PSYCHOTRIA PUNCTATA (RUBIACEAE): GROWTH AND ROOT FORMATION IN CALLUS CULTURES

Attempts were made to obtain bacteria-free plants of Psychotria punctata from tissue cultures. Stem explants and callus derived from them were induced to form roots but failed to form buds on Linsmaier and Skoog medium and 96 chemical modifications of it, including most of those known to induce bud formation in other species. Roots formed with ample IAA (2 mg/liter or more) and a low kinetin concentration (0.25 or 0.50 mg/liter). Adenine inhibited root formation in these media, but tyrosine did not. Tyrosine did lower the percentage of calluses commencing growth. When enzyme-hydrolyzed lactalbumin (1.3 g/liter), kinetin (0.5 mg/liter) and IAA (5 mg/liter) were added to Linsmaier and Skoog medium modified by decreasing inorganic nitrogen and increasing inorganic phosphate, callus grew at the fastest rate observed (increasing threefold in fresh weight in three weeks) and formed numerous roots. This was adopted as the stock callus medium. Casein hydrolysates also stimulated growth but less so than lactalbumin hydrolysate. When lactalbumin hydrolysate or a casein hydrolysate lacking tryptophan was supplied, growth occurred without added auxin if sufficient cytokinin was added. Cytokinin was required at unusually high concentration and was tolerated at still higher concentration. Formation, elongation, and branching of roots persisted on a saturated solution of BA which inhibited callus growth about 70 % and delayed callus senescence. Light caused earlier callus senescence after growth had ceased but did not affect callus growth or root formation. Light-induced senescence was prevented by a high cytokinin concentration.     

江鲽在卵和卵黄囊期仔鱼发育阶段生化成分的变化

本文研究和测定了江鲽( Platichthys flesus L.)在卵和卵黄囊期仔鱼发育阶段水分、钠、钾、脂肪和蛋白质的含量变化。含水量在卵和仔鱼发育期基本保持在90—92%,在仔鱼进入初次摄食期时可降到89%左右。初孵仔鱼具一大卵黄囊,含水量高达93.3%,起“浮力器官”(buoyancy organ)的作用。钠、钾离子含量在卵受精和仔鱼出膜后波动剧烈,呈“上升—下降—再上升”式型。脂肪和蛋白质含量在卵和仔鱼早期发育和饥饿期均呈明显线性下降。同时,本文还就生化成分变化和海洋浮性鱼卵及其仔鱼的生态习性的关系作了初步探讨。     

MORPHOLOGICAL AND BIOCHEMICAL CHANGES IN RAT SYNAPTOSOME FRACTIONS DURING NEONATAL DEVELOPMENT

A biochemical and quantitative morphologic study of presynaptic endings during postnatal development was carried out in subcellular fractions from cerebral cortex of 1, 4, 8, 12, and 18 day old and adult rats. Crude mitochondrial fractions were subfractionated in Ficoll gradients and all resulting fractions were examined in the electron microscope. Presynaptic terminals and other intact processes were counted. Protein content and enzyme activities were assayed in the fractions and in total brain homogenate. In the first and fourth day of life, most of the presynaptic terminals were found in two "light" fractions, between supernatant and 7.5% Ficoll, where they accounted, respectively, for 6 and 22% of all the processes. Progressively with age, more presynaptic terminals were found in the traditional "synaptosomal" fractions between 7.5 and 13% Ficoll. In that region of the gradient, 40, 54, 75, and 89% of the processes were presynaptic endings at 8, 12, and 18 postnatal days and in the adult animal, respectively. A similar shift from the lighter to the heavier fractions was observed in the distribution of choline acetyltransferase and acetylcholinesterase between days 8 and 12. The rate of increase of the specific activity of these two enzymes paralleled that of the percentage of the presynaptic endings after day 8. This study indicates that subcellular fractions can be used to study formation and maturation of synapses during postnatal development.     

ISOENZYME COMPLEMENTS OF DIANTHUS CALLUS CULTURES: INFLUENCE OF LIGHT AND TEMPERATURE

Callus cultures isolated from the stems of two clones of Dianthus were extracted for water-soluble proteins and the extracts were subsequently analyzed by acrylamide gel electrophoresis. Peroxidase, acid phosphatase, esterase, and 6-P-gluconate dehydrogenase (6-P-G DH) were studied under each of four combinations of light and dark, cold (0-5 C) and warm (25 C) environments. Marked differences in the isoenzyme patterns were observed under the four conditions with all the enzymes except 6-P-G DH. Acid phosphatase generally showed fewer isoenzymes than the usual stem complement, whereas the other enzymes showed the same or greater number of isoenzymes. Many of the isoenzymes from the callus tissue correlated well with those found in the stem; however, all but 6-P-G DH showed some different components. Of the four enzymes studied only peroxidase was recovered from the medium; these isoenzymes matched those found in the callus and in the stem. The isoenzymes in the medium were not influenced qualitatively by the growth conditions. The changes in the zymograms of the callus under the four environments were compared to the changes observed in the stem under stress environments.     

MORPHOGENESIS OF POLLEN CALLUS CULTURES OF HYOSCYAMUS NIGER

Morphogenesis of calluses of single pollen grain origin shed from anthers of Hyoscyamus niger cultured in a liquid medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) was followed upon their transfer to a solid medium with or without 2,4-D. In a solid medium lacking 2,4-D, small calluses consisting of one to five nodular groups of cells at the time of inoculation differentiated root and shoot systems and formed miniature seedlings. In the same medium large calluses with several nodules initially formed a crop of bipolar somatic embryoids with well-defined root and shoot axes which subsequently differentiated into seedlings. Irrespective of their size at the time of transfer, calluses grown in a solid medium containing 2,4-D continued to proliferate without showing signs of organogenesis or embryogenesis.     

在离体条件下拐芹(Angelica polymorpha)形成胚性愈伤组织和非胚性愈伤组织过程中几种内源激素的变化(英文)

以拐芹(Angelica potymorpha)的叶柄为外植体,在附加2,4-D(1mg/L) KT(0.5mg/L)MS培养基上诱导脱分化,40天内可形成愈伤组织。在相同的培养基上经过继代培养,可在20天内形成胚性愈伤组织(Embryogenic Callus,EC)和非胚性愈伤组织(Non-Embryogenic Callus,NEC)。将不同天数的培养物作为样品,提取其内源激素;气液相色谱(Gas-Liquid-Chromatography,GLC)测定结果发现,内源ABA,GA_3,CTK和IAA的水平都有变化,尤其是ABA,GA_3。这些结果表明内源ABA和GA_3在胚性愈伤组织和非胚性愈伤组织的分化过程中起着重要作用。     

黄鳝孵卵泡的生化成分及其生理作用

通过对繁殖季节黄鳝为孵卵所吐的泡沫的特性和生化成分分析 ,孵卵泡的黏度为 2 72mPaS ,证明了该孵卵泡是糖蛋白 ,其中糖类含量为 0 15 6mg/mL ,蛋白质含量为 0 2 5 0mg/mL ,蛋白质 /糖类比值为 1 6 0。分离的泡沫蛋白经SDS PAGE电泳含 4种组分 ,分子量从小到大依次为 4 2 5kDa、5 8 6kDa、6 5 3kDa、98 2kDa。泡沫蛋白含 16种氨基酸 ,氨基酸总含量为 2 0 1 88mg/ 10 0mL。同时对孵卵泡的生理作用进行了分析和讨论