Dihydroconiferyl alcohol as a gibberellin synergist in inducing lettuce hypocotyl elongation. An assessment of structure-activity relationships

Structure-activity relationships of the cotyledon factor wereexamined by testing the effect of various substances structurallyrelated to the cotyledon factor (dihydroconiferyl alcohol) ongibberellin-induced lettuce hypocotyl elongation. The biological activity of the cotyledon factor, 3-(4-hydroxy-3-methoxyphenyl)propan-1-ol, disappeared if the phenolic hydroxy group was maskedwith a methoxy or glucosyl group. Oxidation of the alcoholicgroup in the side chain to a carboxylic group decreased thebiological activity of the cotyledon factor. As to relationshipsbetween the biological activity and length of the alkyl sidechain, the propane type was found to be much more active thanthe methane, ethane or butane type. The presence of a C = Cbond in the alkyl side chain made the cotyledon factor biologicallyinactive. Some antioxidants of indole-3-acetic acid were alsoassayed for cotyledon factor-like activity, since the cotyledonfactor is a polyphenol. However, known antioxidants such asrutin, pyrocatechol, chlorogenic acid, caffeic acid and ferulicacid did not show cotyledon factor-like activity. From these results, structural requirements of the cotyledonfactor as a gibberellin synergist were discussed. (Received June 17, 1975; )     

南瓜种子萌发及子叶发育时谷氨酰胺合成酶和其它氨同化酶的变化

在发育的新生组织中 ,来自种子胚乳储存蛋白的降解和氨基酸分解代谢产生的氨由谷氨酰胺合成酶 ( Glutamine synthetase,GS)重新同化 ,生成的谷氨酰胺 ( Gln)被转运到正在生长着的部分。GS是高等植物氮素代谢的关键酶 [1] ,这个酶能同化不同来源的氨。 GS有多种同工酶 ,存在于植物的各种组织和器官中。它们是由一小的同源但分离的核基因家族编码的 [2 3 ] ,这些不同的 GS在植物氮素同化中起着非重叠的作用 [4] ,它们的表达受到环境、发育进程以及组织或细胞类型等许多因素的影响。在大多数已研究过的植物叶片中存在两种 GS,即胞液型GS(…     

Comparative Study on the Bioassay between N6, O2'-Dibutyryl Cyclic Adenosine 3,5-Monophosphate and Cytokinin

The physiological activity of diBcAMP by various classical bioassay methods of cytokinin was studied. It exhibits the cytokinin-like activity in many ways of biological response, such as: the unfolding of etiolated barley leaf, the expansion of radish cotyledon, the expansion and greening of cucumber cotyledon and the inhibition of the elongation of mung-bean hypocotyl, etc. The activity of 1 mM diBcAMP was approximately the same as the activity of 0.044 mM benzyladenine in the most of the tests. However, the activity of 1 mM diBcAMP was found to be higher than the optimal concentration (0.044 mM) of benzyledenine in the radish cotyledon under the expansion test. There was no toxicity of diBcAMP in all of the bioassay methods used, even with higher concentration.     

Sequential Expression of Trypsin Inhibitors in Developing Fruit of Cowpea (Vigna unguiculata (L.) Walp)

Trypsin inhibitor (TI) activity was followed in the pod (pericarp),seed coat, cotyledon and embryo axis during fruit developmentof cowpea. On the basis of seed fresh weight, three phases couldbe distinguished from anthesis to fruit maturity. In the podTI activity increased from the beginning of Phase I to a maximumin the middle of the phase. From then on the activity declineduntil no activity could be detected before the end of phaseII. The cotyledons did not contain any TI in Phase I. TI activitywas first detected in the cotyledon in the beginning of PhaseII at the same time that globulin synthesis started. The TIactivity in the cotyledon increased to a maximum at the endof Phase II before decreasing in Phase III. In the embryo axisa similar pattern of TI activity to that of the cotyledon wasfound. No protein TI could be detected in the seed coat at anystage. In the pod there is a TI with a mol. wt of 12500 andpI of 4.4. Mature cotyledon and embryo axis have two TI withmol. wt 10800 and 24700 with pI 4.7 and 5.0 respectively. Duringdevelopment the smaller TI (mol. wt 10800) was synthesised beforethe larger TI (mol. wt 24700). There were large differencesbetween the maximum absolute amounts of TI present in the pericarp,cotyledon and embryo axis.     

豌豆种子萌发时的含水量对子叶中蛋白酶和淀粉酶活性的影响

不同含水量的豌豆种子在饱和水蒸气中保持7d,在此过程中,当含水量低于萌动临界含水量时,限制了子叶中淀粉酶长寿mRNA的转译;含水量达到或超过萌动临界含水量时,子叶中淀粉酶长寿mRNA的转译得以进行,但转译程度因含水量不同而异,正常生长的轴器官是吸收子叶中蛋白质和淀粉水解产物的动力库,子叶中氨基酸含量的变化灵敏地调节着蛋白酶的活性,从而影响着蛋白质的动员程度,当环境中供水不足时,发现子叶中还原糖积累很快,非还原糖含量几乎没有变化,因此,认为对淀粉酶活性起灵敏调节作用的主要是还原糖。     

Purification, properties and alternate substrate specificities of arginase from two different sources: Vigna catjang cotyledon and buffalo liver

Arginase was purified from Vigna catjang cotyledons and buffalo liver by chromatographic separations using Bio-Gel P-150, DEAE-cellulose and arginine AH Sepharose 4B affinity columns. The native molecular weight of an enzyme estimated on Bio-Gel P-300 column for Vigna catjang was 210 kDa and 120 kDa of buffalo liver, while SDS-PAGE showed a single band of molecular weight 52 kDa for cotyledon and 43 kDa for buffalo liver arginase. The kinetic properties determined for the purified cotyledon and liver arginase showed an optimum pH of 10.0 and pH 9.2 respectively. Optimal cofactor Mn++ ion concentration was found to be 0.6 mM for cotyledon and 2 mM for liver arginase. The Michaelis-Menten constant for cotyledon arginase and hepatic arginase were found to be 42 mM and 2 mM respectively. The activity of guanidino compounds as alternate substrates for Vigna catjang cotyledon and buffalo liver arginase is critically dependent on the length of the amino acid side chain and the number of carbon atoms. In addition to L-arginine cotyledon arginase showed substrate specificity towards agmatine and L-canavanine, whereas the liver arginase showed substrate specificity towards only L-canavanine.     

Purification and properties of pea cotyledon and embryo diamine oxidase

Diamine oxidase was purified separately from cotyledon and embryo of pea seedlings germinated for 6 days. The K_m of the cotyledon enzyme for putrescine was 1.6 × 10^?4M while that for the embryo enzyme was 9 × 10^?5M. On heating for 15 min at 70° the embryo enzyme retained about 90% activity whereas the cotyledon enzyme retained only 20% activity. The electrophoretic mobility of the cotyledon enzyme was ca twice that of the enzyme from embryo.     

Ultrastructure and Lipase Activity of Cotyledon cell During Pod Development in Peanut

A series of significant changes of the ultrastructure and lipase activity of cotyledon cell were found in peanut (Arachis hypogaea) during pod development. In he initial stage of cotyledon development there were many plastids which kept producing starch grain and there were low lipase activity and very few lipid and protein bodies in the cell. In the middle stage of cotyledon development, a great number of larger lipid bodies were seen in the cell and a lot of protein bodies formed in the vacuoles and continued to increase in size. Lipase activity increased in the cytoplasm, endoplasmic reticulum, protein bodies, plasmalemma and intercellular space. In the later stage of cotyledon development, the lipid bodies did not increase in number but became slightly larger. The protein bodies continued to increase both in number and in size. Lipase acttvity was even hegher in the cytoplasm. In the final stage the protein bodies became irregular in shape and some of them tended to disintegrate with their content entered into the space around the lipid bodies. The lipase activity in the cell declined. The results indicated that the lipid body originated in the cytoplasm and the protein body originated in the vacuole; that the accumulation of oil and protein in peanut cotyledon resulted from the formation and development of lipid and protein bodies in the cell, and that the changes of plasmid and lipase activity in the cell played a role in the development of lipid body during the development of cotyledon.     

Metabolism of Inositol Phosphates: I. Phytase Synthesis during Germination in Cotyledons of Mung Beans, Phaseolus aureus

The degradation of phytin in germinating mung bean seeds has been found to be associated with the increased activity of phytase in the cotyledon. In the differentiated embryo the increase of this activity is very low all throughout the growth periods studied. Phytase appears in the cotyledon during germination. No activity has been detected in the cotyledons of unsoaked seeds. Cycloheximide (10⁻⁶ M) inhibits the appearance of phytase by 61% during 24 and 48 hours after the start of germination. This phytase increase is dependent on the synthesis of new RNA in the cotyledon. Synthesis of DNA is not detected in the cotyledon during germination.     

ATPase Activity of Pea Cotyledon Submitochondrial Particles: ACTIVATION, SUBSTRATE SPECIFICITY, AND ANION EFFECTS

Submitochondrial particles freshly prepared by sonication from pea cotyledon mitochondria showed low ATPase activity. Activity increased 20-fold on exposure to trypsin. The pea cotyledon submitochondrial particle ATPase was also activated by “aging” in vitro. At pH 7.0 addition of 1 millimolar ATP prevented the activation. ATPase of freshly prepared pea cotyledon submitochondrial particles had a substrate specificity similar to that of the soluble ATPase from pea cotyledon mitochondria, with GTPase > ATPase. “Aged” or trypsin-treated particles showed equal activity with the two substrates. NaCl and NaHCO₃, which stimulate the ATPase but not the GTPase activity of the soluble pea enzyme, were stimulatory to both the ATPase and GTPase activities of freshly prepared submitochondrial particles. However, they were stimulatory only to the ATPase activity of trypsin-treated or “aged” submitochondrial particles. In contrast, the ATPase activity of rat liver submitochondrial particles was stimulated by HCO₃⁻, but inhibited by Cl⁻, indicating that Cl⁻ stimulation is a distinguishing property of the pea mitochondrial ATPase complex.     

发育和早萌中花生胚胚轴与子叶内BAPAase活性的差异

花生胚发育过程中,子叶和胚轴中都出现BAPAase活性。花生种子萌发时,子叶和胚轴中的BAPAase活性迅速上升,子叶中无新的BAPAase合成,胚轴中能重新合成BAPAase。ABA抑制了子叶和胚轴中BAPAase的活性,抑制胚轴中BAPAase活性所需的外源ABA浓度更高,Act-D和CHM不能逆转ABA对BAPAase活性的抑制作用。     

The Changes of H+-ATPase During the Mitochondrial Development of Pea Cotyledon

Electron microscopic observation indicated that the mitochondrial membrane of pea cotyledon gradually developed into integral structure during seeds imbibition. ATP-synthesizing activity of H+-ATPase increased in company with mitochondrial development, but the content of F1-ATPase subunits was not different on the mitochondria of cotyledon imbibed for 6 hours and for 24 hours in water. After cotyledon was imbibed at low temperature, the content of γ and β subunits of F1-ATPase was distinctly reduced with the inhibition of H+-ATPase activity.     

花生荚果发育过程中子叶细胞的超微结构和脂酶活性的研究

在花生(Arachis hypogaea)荚果发育过程中,子叶细胞的超微结构和脂酶活性皆发生了显著变化。子叶生长初期,缅胞中质体较多,并不断形成淀粉粒;脂酶活性低,脂体和蛋白体很少。子叶发育中期,子叶细胞质中出现大量体积较大的脂体,液泡中的蛋白体不断形成和增大,而且细胞质、内质网、蛋白体外膜、细胞质膜和细胞间隙上皆显示较强的脂酶活性。子叶发育后期,脂体数量不再增加,但体积略有增大,间质透明度也有提高;蛋白体增大较小,但数量却进一步增多;细胞质中仍显示较强的脂酶活性。至末期时,蛋白体形态变得不规则,甚至出现部分解体,其基质充挤脂体间隙;细咆中的脂酶活性减弱。研究表明,花生脂体起源于细胞质,蛋白体起源于液泡,子叶油分和蛋白质的积累足体内脂体和蛋白体不断发育的结果,细胞中脂酶活性的变化可能与脂体发育有关。     

Seed-specific overexpression of a potato sucrose transporter increases sucrose uptake and growth rates of developing pea cotyledons

During the storage phase, cotyledons of developing pea seeds are nourished by nutrients released to the seed apoplasm by their maternal seed coats. Sucrose is transported into pea cotyledons by sucrose/H+ symport mediated by PsSUT1 and possibly other sucrose symporters. PsSUT1 is principally localised to plasma membranes of cotyledon epidermal and subepidermal transfer cells abutting the seed coat. We tested the hypothesis that endogenous sucrose/H+ symporter(s) regulate sucrose import into developing pea cotyledons. This was done by supplementing their transport activity with a potato sucrose symporter (StSUT1), selectively expressed in cotyledon storage parenchyma cells under control of a vicilin promoter. In segregating transgenic lines, enhanced [(14)C]sucrose influx into cotyledons above wild-type levels was found to be dependent on StSUT1 expression. The transgene significantly increased (approximately 2-fold) transport activity of cotyledon storage parenchyma tissues where it was selectively expressed. In contrast, sucrose influx into whole cotyledons through the endogenous epidermal transfer cell pathway was increased by only 23% in cotyledons expressing the transgene. A similar response was found for rates of biomass gain by intact cotyledons and by excised cotyledons cultured on a sucrose medium. These observations demonstrate that transport activities of sucrose symporters influence cotyledon growth rates. The attenuated effect of StSUT1 overexpression on sucrose and dry matter fluxes by whole cotyledons is consistent with a large proportion of sucrose being taken up at the cotyledonary surface. This indicates that the cellular location of sucrose transporter activity plays a key role in determining rates of sucrose import into cotyledons.     

Phytochrome-induced intercellular signalling activates cab::luciferase gene expression

The phytochrome-induced expression pattern of the chlorophyll a/b binding protein (cab) gene was studied using a cab2::luciferase reporter transgene in the tobacco cotyledon. The role of developmentally regulated competence, cooperativity among cells and signal propagation was investigated by red-light microbeam irradiation of distinct areas of the cotyledon. Even with a minimal fluence, the response was not restricted to the irradiated cells. Following irradiation of the cotyledon base, the luciferase activity revealed a robust propagation of activating signals to areas that had not received a light stimulus. The acropetal outspread formed distinct expression patterns depending on the site of the irradiation. The combination of imaging luciferase activity in living seedlings and microbeam microscopy provides significant experimental evidence of how cellular light perception and intercellular signalling contribute to the cab gene expression pattern.     

Chromatin-bound DNA-dependent RNA polymerase in developing pea cotyledons

Summary The pattern of cotyledon development in three varieties of Pisum sativum has been defined in terms of cell number, DNA and RNA content and chromatin, bound RNA polymerase activity. Variation was observed in the relative periods of growth by cell division and cell expansion between the three varieties. The mean DNA content per cotyledon cell during growth by cell expansion increased to approximately 50C in one variety, 30C in the second variety and 15C in the third variety. The pattern of chromatin-bound RNA polymerase activity during development suggested that some of the DNA above the 2C level may contribute to RNA synthesis in two of the three varieties studied. In the third variety the RNA polymerase activity decreases throughout the phase of increase in DNA per cell. The chromatin-bound RNA polymerase activity per cell was correlated with the rate of RNA increase per cell.     

In Vitro Stability and Inactivation of Peptide Hydrolases Extracted from Phaseolus vulgaris L.

Endopeptidase activity against azocasein had a higher temperatureoptimum (50°C) in leaf extracts than in cotyledon extracts(37°C). The temperature optima for aminopeptidase (46°C)and for carboxypeptidase (53°C) were similar in leaf andcotyledon extracts. The endopeptidase activity showed an excellentstability in crude extracts from leaves even at 37°C, whilethe endopeptidase in cotyledon extracts was less stable. Carboxypeptidasewas very stable in both leaf and cotyledon extracts. Aminopeptidasewas the least stable of the enzymes investigated and its inactivationrate depended on the source of the extract. A moderate stabilitywas observed in extracts of leaves or of ungerminated seeds,but this enzyme was rapidly inactivated in cotyledon extractsat pH 5.4. At pH 7.5 aminopeptidase remained active longer thanat pH 5.4. From experiments with mixed extracts it could beconcluded that in cotyledons an aminopeptidase inactivatingfactor was formed during germination. This factor was heat sensitive,excluded by Sephadex G-25, precipitated by 75% ammonium sulfateand inhibited by tosyl-L-lysine chloromethyl ketone. These datasuggest that the factor is a protein and considering the similarproperties it appears possible that it is the endopeptidaseformed during germination. (Received May 15, 1981; Accepted July 18, 1981)     

Ultrastructural Localization of Adenosine Triphosphatase Activity in Cotyledon Cells of Tomato and Its Changes during Chilling Stress

The ultrastructural localization of adenosine tripkosphatase (ATPase) activity in cotyledon cells of tomato was carried out by use of the cytochemical method of lead phosphate precipitation, and the changes in ATPase activity during chilling stress of the tomato seedlings were studied. The following experimental results have been obtained: 1. The ATPase activity in the cotyledon cells of tomato seedlings germinated and grown at 28 ℃. was located at plasmolemma, plasmodesmata, nucleoli and nuclear chromatin chloroplast lamellae, many sites of cell wall, and the surface of cell wall bordering the intercellular spaces and their inclusions. 2. When the tomato seedlings were subjected to chilling treatment for 4 h. at 5 ℃., the ATPase activity in cotyledon cells was indifferent from that of non-chilling treated seedlings. After chilling treatment for 12 h. at 5 ℃., the reaction of ATPase activity at plasmolemma, and in cell wall and intercellular spaces was markedly reduced. though the high activity reaction of ATPase in nuclei and at chloroplast lamellae was still maintained. When the tomato seedlings were subjected to chilling stress for 24 h. at 5℃., the ATPase activity at plasmolemma and in cell wall was almost inactivated, while the ATPase activity in nuclei and at chloroplast lamellae was only slightly lowered. These results indicated that the chilling injury may influence firstly on the ATPase activity of cell surface (plasmolemma and cell wall). 3. The role of intercellular spaces used as the passage of materials and the process and mechanism of chilling injury are discussed.     

Histochemical studies on protease formation in the cotyledons of germinating bean seeds

Summary Protease formation in Phaseolus vulgaris L. cotyledons during seed germination was studied histochemically using a gelatin-film-substrate method. Protease activity can be detected by this method on the 5th day of germination, at approximately the same time that a rapid increase of activity was observed by a test-tube assay with casein as a substrate. At the early stage of germination, protease activity was observed throughout the cotyledon except in two or three cell layers below the cotyledon surface and in several cell layers around the vascular bundles. A highly active cell layer surrounding the protease-inactive cells near the vascular bundles is suggested to be a source of the protease.Brooklyn Botanic Garden Contribution No. 202.