Lycopene induction in Cucurbita moschata cotyledons by CPTA

2-(4-Chlorophenylthio) triethylamine hydrochloride (CPTA) inducedthe rapid accumulation of lycopene in Cucurbita moschata cotyledons.Concomitant with this increase, was a decrease in chlorophyllconcentration in green seedlings, a cessation of further plumuleand cotyledonary growth and an increase in cotyledon respiration.Light was required for maximum lycopene accumulation. (Received August 4, 1973; )     

The metabolism of proline in cotyledons of pumpkin (Cucurbita moschata)

Exogenous proline-U-₁₄C is readily metabolized to glutamate,ornithine, sugars, CO₂, and organic acids, and is incorporatedinto protein by etiolated and green pumpkin cotyledons. As littletranslocation of proline from the cotyledons occur, it was proposedthat in young tissue proline is converted to glutamate, ornithineor sugar which are then readily translocated from the cotyledons.In older tissue some glutamate carbon derived from proline isalso used as an energy source and metabolized to CO₂. As proteinsynthesis is occurring rapidly in these cotyledons, considerableproline is incorporated into new protein. After 10-hr, 15% ofthe absorbed radioactivity still remained as free proline. ¹Present address: Instituto de Ciencias Biologicas, UniversidadeFederal de Vicosa, Vicosa, Minas Gerais, Brasil. (Received February 1, 1974; )     

Pigment synthesis in Cucurbita moschata cotyledons as influenced by CPTA and several inhibitors

2-(4-Chlorophenylthio) triethylamine hydrochloride (CPTA) inducedthe accumulation of a number of carotenes in pumpkin (Cucurbitamoschata) cotyledons, a tissue which does not normally accumulatethese pigments. Lycopene accumulated concomitantly with a decreasein -carotene and ß-carotene. CPTA appeared to inhibitcyclases involved in the synthesis of -carotene and ß-caroteneand to stimulate enzymes involved in lycopene synthesis. Cycloheximidereduced this CPTA induced enzyme synthesis while diphenylaminereduced CPTA induced carotene accumulation. Actinomycin D alonereduced accumulation of carotenes, but it did not affect CPTAinduced carotene accumulation. Rather lutein, violoxanthin andneoxanthin decreased and -carotene and ß-caroteneaccumulated. ¹Present address: Morioka Branch, Vegetable and Ornamental CropsResearch Station, Ministry of Agriculture and Forestry, Shimokuriyagawa,Morioka, Japan. (Received August 12, 1974; )     

Proline-Dehydrogenase from Pumpkin (Cucurbita moschata) Cotyledons

A NAD specific proline-dehydrogenase was found in pumpkin (Cucurbita moschata Poir. cv. Dickinson Field) which oxidized proline to Δ¹-pyrroline-5-carboxylate. NADP did not substitute for NAD and L-proline-methyl-ester and thiazolidine-4-carboxylate were substrates in the reaction, at a rate of 107% and 33% respectively, of the rate with L-proline. Pumpkin cotyledons contained the bulk of the enzyme activity with 90% of the activity being in the soluble fraction. Proline-dehydrogenase, which was not treated at high temperature, was stable at –10°C for 4 months in the presence of high ammonium sulfate concentration. The Michaelis constant for NAD was 2.2 mM and for L-proline was 2.5 mM. At 5 mM NADP, a 40% non-competitive inhibition of proline-dehydrogenase was obtained, while 50 μM NADP was sufficient to induce 20% inhibition.     

大量元素胁迫下南瓜化感作用研究

研究采用水培实验、生物测试及室内分析相结合的方法,研究在不同大量元素(N、P、K、Ca、S、Mg)营养胁迫下幼苗期南瓜根系分泌物及其提取物对南瓜幼苗生长的化感作用。结果表明：南瓜具有自毒作用,不同大量营养元素胁迫对其胚根和胚芽生长均产生了抑制作用。不同的营养条件对其根系分泌物的影响不同,营养元素的合理搭配可以减轻自身的自毒作用。南瓜根系分泌物经乙酸乙酯提取的活性有机物埘南瓜胚根牛长亦表现出抑制作用,且抑制作用程度较大。南瓜幼苗期根系十物质水浸液巾存在着抑制南瓜幼苗生长的化感物质：筛选出南瓜根系分泌物对自身幼苗生长产生抑制作用最弱的大量元素水平优化组合,为南瓜的营养胁迫机理提供依据。     

南瓜单粒花粉可溶蛋白等电聚焦研究

利用显微操作方法对南瓜单个花粉可溶蛋白成功地进行了等电聚焦,获得了清晰的电泳谱带。分析其谱带表明,同一植株同一朵花上的花粉之间存在着蛋白谱带的差异,统计结果显示其按１：１的孟德尔规律分离。单花粉蛋白等电聚焦结合显微镜操作技术对于遗传育种和发衣生物学研究具有重要的应用前景。     

南瓜组培根根系分泌物的化感效应研究

使用组织培养技术、生物测试法及室内分析相结合的方法研究了南瓜组培根在不同生长时期与不同营养谐迫下(利用正交设计调节标准B₅培养基中的大量元素、微量元素、有机质和激素)根分泌物对南瓜、萝卜和小麦3种受体的种子萌发及幼苗生长的化感作用.结果表明,南瓜具有自毒作用,南瓜组培根根系分泌物对受体幼苗生长的抑制作用呈现出高抑低促的作用表型;南瓜组培根根系分泌物产生化感作用的活性与南瓜组培根生长速度有关,南瓜组培根根系分泌物的生长抑制活性以15～17d即生长速度指数末期为最高,21d生物量最大时的南瓜组培根根系分泌物的生长抑制活性最低;营养元素的改变明显影响了南瓜根系分泌物的产生,并通过筛选出能诱导南瓜组培根对受体具有强烈抑制作用的大量元素、微量元素、维生素和激素种类和含量的最优化组合,为研究南瓜化感作用营养胁迫机理提供依据.     

Alanine aminotransferase and glycine aminotransferase from maize (Zea mays L.) leaves.

Alanine aminotransferase (AlaAT, EC 2.6.1.2) and glycine aminotransferase (GlyAT, EC 2.6.1.4), two different enzymes catalyzing transamination reactions with L-alanine as the amino-acid substrate, were examined in maize in which alanine participates substantially in nitrogen transport. Preparative PAGE of a partially purified preparation of aminotransferases from maize leaves gave 6 fractions differing in electrophoretic mobility. The fastest migrating fraction I represents AlaAT specific for L-alanine as amino donor and 2-oxoglutarate as amino acceptor. The remaining fractions showed three aminotransferase activities: L-alanine-2-oxoglutarate, L-alanine-glyoxylate and L-glutamate-glyoxylate. By means of molecular sieving on Zorbax SE-250 two groups of enzymes were distinguished in the PAGE fractions: of about 100 kDa and 50 kDa. Molecular mass of 104 kDa was ascribed to AlaAT in fraction I, while the molecular mass of the three enzymatic activities in 3 fractions of the low electrophoretic mobility was about 50 kDa. The response of these fractions to: aminooxyacetate, 3-chloro-L-alanine and competing amino acids prompted us to suggest that five out of the six preparative PAGE fractions represented GlyAT isoforms, differing from each other by the L-glutamate-glyoxylate:L-alanine-glyoxylate:L-alanine-2-oxoglutarate activity ratio.     

Glutamine synthesis in germinating seeds of Cucurbita moschata

During germination, an increase in glutamine and glutamine synthetase[L-glutamate: ammonia ligase (ADP), EC 6.3.1.2 [EC] ] occurred inthe cotyledons reaching a maximum at 4 to 6 days. The enzymehad a Km of 4.5 nun for L-glutamate, and 0.67 mu for hydroxylamine.Hydroxylamine exhibited substrate inhibition kinetics. The enzymewas inhibited by calcium ion, fluoride ion and p-hydroxymercuribenzoatebut not by EDTA, sodium ion or chloride ion. The sulf hydrylinhibition was reversed by dithiothreitol. In vivo synthesisof glutamine-¹⁴C from glutamate-¹⁴C was found to parallel theincrease in glutamine content and the in vitro glutamine synthetaseactivity during germination. ¹ Present address: Department of Biology, Mercyhurst College,Erie, Pennsylvania 16501, U.S.A. (Received June 12, 1971; )     

Isolation of serine:glyoxylate aminotransferase from cucumber cotyledons

Serine:glyoxylate aminotransferase, a marker enzyme for leaf peroxisomes, has been purified to homogeneity from cucumber cotyledons (Cucumis sativus cv Improved Long Green). The isolation procedure involved precipitation with polyethyleneimine, a two-step ammonium sulfate fractionation (35 to 45%), gel filtration on Ultrogel AcA 34, and ion exchange chromatography on diethylaminoethyl-cellulose, first in the presence of pyridoxal-5-phosphate, and then in its absence. The enzyme was purified approximately 690-fold to a final specific activity of 34.4 units per milligram. Electrophoresis of the purified enzyme on sodium dodecyl sulfate-polyacrylamide gels revealed two polypeptide bands with apparent molecular weights of approximately 47,000 and 45,000. Both polypeptides coeluted with enzyme activity under all chromatographic conditions investigated, both were localized to the peroxisome, and both accumulated in cotyledons as enzyme activity increased during development. The two polypeptides appear not to be structurally related, since they showed little immunological cross-reactivity and gave rise to different peptide fragments when subjected to partial proteolytic digestion. Antiserum raised against either the denatured enzyme or the 45,000-dalton polypeptide did not react with any other polypeptides present in a crude cotyledonary homogenate. The purified enzyme also had alanine:glyoxylate aminotransferase activity, but was about twice as active with serine as the amino donor.     

南瓜品质资源的分析测试

本文对南瓜不同品种进行营养成分的分析和比较 ,为南瓜品质资源的筛选提供一定的实验依据 ,为南瓜作为蔬菜、加工或观赏提供恰当的品种     

Glucose-lowering activity of novel tetrasaccharide glyceroglycolipids from the fruits of Cucurbita moschata

Two new tetrasaccharide glyceroglycolipids were obtained from pumpkin. The structures of the two compounds were determined using chemical methods and spectroscopic analysis. Both compounds demonstrated significant glucose-lowering effects in streptozotocin- and high-fat-diet-induced diabetic mice.     

Light and chlorophyll gradients within Cucurbita cotyledons

Abstract. Measurement of light within 10–14-d-old green and etiolated Cucurbita pepo cotyledons were made with fibre-optic microprobes to assess the influence of chlorophyll distribution and anatomical variations in mesophyll cell type (spongy versus palisade) on internal light pattern. More than 50% of the pigment in green cotyledons occurred in the upper (adaxial) 300 μm and this gradient strongly influenced the internal propagation of 680 nm light. When the upper (adaxial) surface was irradiated with 680 nm light, almost complete absorption occurred within the first 400 μm (palisade) of approximately 1200-μm-thick cotyledons. In contrast, when lower (abaxial) surfaces were irradiated with 680 nm light, penetration extended throughout the spongy mesophyll to about the 700 μm depth. Measurements of collimaled and scattered light gradients at 550, 680 and 750 nm indicated that collimaled light was rapidly scattered by mesophyll cells. In cotyledons irradiated on the upper surface, spongy mesophyll cells received only scattered light. Furthermore, comparisons of scattered light gradients obtained from cotyledons irradiated on upper and lower surfaces suggested that spongy mesophyll cells scatter light more effectively than palisade cells, probably due to the greater proportion of intercellular air spaces in spongy mesophyll tissue. These data also indicate that both the spectral quality and quantity of light incident on palisade versus spongy mesophyll cells differs, perhaps contributing to developmental and physiological differences between these two mesophyll cell types.     

Location of gluconeogenesis from phosphoenolpyruvate in cotyledons of Cucurbita pepo.

1. The aim of this work was to discover the location of the enzymes that convert phosphoenolpyruvate to fructose 6-phosphate during gluconeogenesis in fatty seeds. Cotyledons of 5-day-old dark-grown seedlings of marrow (Cucurbita pepo) were used as experimental material. 2. Cotyledons were separated into palisade and mesophyll tissue. Extracts of the two tissues had comparable activities of gluconeogenic enzymes. 3. Extracts of cotyledons were fractionated by density gradient centrifugation to yeild mitochondria and glyoxysomes, and by gel filtration to yield proplastids. The isolated organelles retained their characteristic ultrastructure and appreciable amounts of marker enzymes. The proportions of the total activities of phosphoglyceromutase and fructose-1, 6-diphosphatase recovered in the mitochondrial and glyoxysomal preparations were insignificant. The same was true for the activities of phosphoglyceromutase and phosphopyruvate hydratase found in the proplastid preparations. 4. Extracts of a number of other gluconeigenic plant tissues were centrifuged at 2500 times g to yield particulate preparations. None of these preparations contained a significant proportion of the total activity of phosphoglyceromutase. 5. It is suggested that gluconeogenesis from phosphoenolpyruvate in plants occurs in the cytoplasm.     

南瓜两性花的形态与结构研究

对南瓜的两性花进行外部形态及结构的研究,结果表明：南瓜的两性花可有子房上位花、子房半下位花和子房下位花三种类型：花萼、花冠均为５,合瓣;雄蕊３,其中有两枚各由两个雄蕊合生而成,分离和部分联合,花药结构特殊,花粉发育正常;雌蕊具有单个或两个柱头及花柱。子房壁结构正常。上方的子房壁表皮发育完好,具气孔器。胚珠在外形上也发育正常。两性花与单性花同生长在一个植株上,可以连续多代稳定地遗传,萌生当代植株的种子来源于上一代雌花。这对探讨被子植物的系统进化关系有着重要的意义     

Protein composition of Cucurbita maxima and C. moschata seeds

Seeds of Cucurbita maxima, C. moschata and their interspecific hybrids were used to evaluate the intrapopulational and interpopulational variation of their protein composition. Three immunoprecipitating systems common to all the studied samples were detected by the Ouchterlony technique. Fourteen protein bands were identified by polyacrylamide gel electrophoresis (PAGE) whereas 23 bands were identified by sodiumdodecylsulfate (SDS)-PAGE. Using Western blotting (WB) also 23 bands were detected. The Jaccard's index of similarity calculated from SDS-PAGE and WB varied between 91 and 100 % for all the compared pairs of samples. These results demonstrate a high uniformity in the protein composition of all the samples and do not allow for their clear characterization.     

南瓜高效再生体系的建立

南瓜(Cucurbita moschata)再生率较低, 为建立高效的南瓜再生体系, 以南瓜子叶为外植体, 进行35组不同激素浓度的不定芽诱导研究。结果表明, 南瓜再生受培养基中激素浓度和配比的影响, 适宜浓度6-苄氨基腺嘌呤(6-BA)能有效促进不定芽形成; 单独使用脱落酸(ABA)诱导使南瓜子叶发黄, 但与6-BA组合使用可显著提高外植体的再生能力, 1.0 mg?L ^-16-BA与0.5 mg?L ^-1ABA组合南瓜芽再生率高达90.26%。将不定芽置于MS培养基中进行生根培养, 再生苗移栽易成活。从子叶接种到苗再生约需70天。     

农杆菌介导南瓜遗传转化体系的建立

以南瓜金辉一号（Cucurbita moschata＇ Jinhui1＇）为实验材料,利用根癌农杆菌（Agrobacterium tumefaciens）介导转化南瓜子叶节,研究了预培养时间、侵染时间、乙酰丁香酮（AS）浓度和共培养时间,抗生素羧苄青霉素（Carb）、头孢霉素（Cef）以及筛选剂卡那霉素（Kan）等因素对离体不定芽的影响,建立了南瓜最适遗传转化体系。结果表明：外植体预培养0天,侵染时间30分钟,AS浓度为100mg·L^-1,共培养5天可获得最高遗传转化效率;最适除菌剂为Cef,其最适浓度为500mg·L^-1;最适Kan筛选浓度为100mg·L^-1;在MS培养基上培养抗性芽生根,经PCR和Southern blot检测,证明为转基因植株。