Über einige weitere harnstofführende Pflanzen

Zusammenfassung Es werden im Anschluß an frühere Untersuchungen fünf neue Befunde über Harnstoffvorkommen bei höheren Pflanzen mitgeteilt, die mit der histochemischen Xanthydrolmethode bzw. mit einer Ureasemethode gewonnen wurden. Harnstoffmengen, die unter der Erfassungsgrenze der Mikroreaktion liegen, lassen sich mit der Ureasemethode noch bestimmen. Die positiven Pflanzen sind:Ornithopus sativus, Linum usitatissimum, Luffa aegyptiaca, Lagenaria vulgaris, Bryonia alba.     

Distribution and morphology of extrafloral nectaries in some Cucurbitaceae

A study of extrafloral nectaries has been made in the Cucurbitaceae to ascertain their structure and assess their taxonomic potential. Nineteen species representing nine Old World genera and one New World genus were examined. These included Telfairia occidentalis, Telfairia pedata, Momordica charantia, Lagenaria siceraria, Citrullus lanatus, Luffa aegyptiaca, Cucurbita moschata and Trichosanthes cucumerina , which are of economic importance and cultivated in Nigeria for their leaves and/or fruits.
Observation of the regularity of ant and insect-visitors, along with tests for glucose and β-glucosidase enzymes, revealed the presence of extrafloral nectaries in nine species. Considerable variation exists in the distribution and morphology of nectaries between genera, especially in the tribe Benincaseae. The nutritional and ecological significance of the occurrence of extrafloral nectaries in Telfairia occidentalis is discussed.     

Purification and characterization of Luffin P1, a ribosome-inactivating peptide from the seeds of Luffa cylindrica

A peptide designated Luffin P1 was purified from the seeds of Luffa cylindrica. Its molecular mass was determined to be 5226.1 Da by MALDI-TOF MS analysis. The purified Luffin P1 shows a strong inhibitory activity on protein synthesis in the cell-free rabbit reticulocyte lysate with IC(50) of 0.88 nM. Its reaction mechanism is the same as that of the ribosome-inactivating protein trichosanthin, which is an rRNA N-glycosidase. Besides, the results of gel filtration chromatography suggested the existence of polymers of Luffin P1 and polymerization of Luffin P1 enhanced its rRNA N-glycosidase activity. Luffin P1 was the smallest peptide yet reported that has translational inhibitory activity. The cDNA and deduced amino acid sequence of Luffin P1 has also been determined.     

丝瓜肌球蛋白的电子显微学研究

从丝瓜(Luffa cylindrica (L.) Roem.)卷须中纯化得到分子量为174kD的肌球蛋白,并对其进行了酶学与电子显微学的研究.这种肌球蛋白具有肌动蛋白激活的MgATPase活性,能够被抗动物肌肉的肌球蛋白的单克隆抗体识别.电子显微学研究表明:它有两个头部(大小和形状与动物肌肉的肌球蛋白相似)和一条相对较短的尾部.还对丝瓜卷须的肌动蛋白进行了观测,偶尔发现一些尾部有球状结构的肌球蛋白.该肌球蛋白的免疫特性和超微结构证明了它由2条重链组成,并与传统的肌球蛋白相似.然而,这种174 kD的肌球蛋白是否参与了丝瓜的接触卷曲有待于进一步研究.     

丝瓜肌球蛋白的电子显微学研究

从丝瓜 (Luffacylindrica (L .)Roem .)卷须中纯化得到分子量为 174kD的肌球蛋白 ,并对其进行了酶学与电子显微学的研究。这种肌球蛋白具有肌动蛋白激活的MgATPase活性 ,能够被抗动物肌肉的肌球蛋白的单克隆抗体识别。电子显微学研究表明 :它有两个头部 (大小和形状与动物肌肉的肌球蛋白相似 )和一条相对较短的尾部。还对丝瓜卷须的肌动蛋白进行了观测 ,偶尔发现一些尾部有球状结构的肌球蛋白。该肌球蛋白的免疫特性和超微结构证明了它由 2条重链组成 ,并与传统的肌球蛋白相似。然而 ,这种 174kD的肌球蛋白是否参与了丝瓜的接触卷曲有待于进一步研究。     

Photosynthetic Prokaryotes. Cell Differentiation and Functions : Edited by G.C. Papageorgiou and L. Packer Elsevier Biomedical; New York, 1983 405 pages. $65.00

A protein (designated as luffin) with an app. M_r of 26000, which inhibits protein synthesis in rabbit reticulocyte lysate, was purified to homogeneity from the seeds of Luffa cylindria roem by extraction with 20 mM Na phosphate buffer (pH 7.2) containing 0.2 M NaCl, ammonium sulfate fractionation, and chromatography on Sephacryl S-200 and CM-Sephadex C-25. Luffin exhibited 10-times as strong inhibitory activity against protein synthesis in rabbit reticulocyte lysate (IC₅₀, 0.42 ng/ml) as that of ricin A-chain, but it showed only a weak cytotoxicity against murine leukemia L1210 cells, an activity of 1/10⁶ to 1/10⁵ that of ricin.     

Larvicidal activity of saponins from Balanites aegyptiaca callus against Aedes aegypti mosquito

Seeking an alternative approach for producing a larvicidal product from Balanites aegyptiaca plants, callus was produced from in vitro cultures of root explants and its larvicidal activity against Aedes aegypti mosquito larvae was evaluated. Concentrations of 0, 50, 100, 500, 1000, and 1500 ppm of saponins from the root-derived callus of B. aegyptiaca were used to determine larvicidal effects and consequent effect on adult emergence. A dose-dependent effect was observed. In a chronic mortality assessment (after 7 days of exposure), concentrations of 500 ppm or greater killed 100% of the test larvae population. Fifty parts per million showed no difference in larval mortality compared to the control (0 ppm); however, this concentration allowed one-fourth of the adult emergence of the control treatment. These results suggest that saponins from in vitro cultures of the root explant of B. aegyptiaca can be used as a larvicidal agent against A. aegypti larvae.     

Multivariate analysis of genetic exchanges between Solea aegyptiaca and Solea senegalensis (Teleosts, Soleidae)

Electrophoretic data from different populations of the teleosts Solea senegalensis, S. aegyptiaca and S. vulgaris were analysed using multivariate analysis. This confirmed that Solea vulgaris is very different genetically and isolated from both senegalensis and aegyptiaca , and that the latter two forms are also well differentiated. The presence of senegalensis alleles in the aegyptiaca population of the Golfe-du-Lion, and reciprocally, aegyptiaca alleles in the senegalensis population of the Gulf of Tunis, suggests that genetic exchange occurs between them. The association of these alleles (gametic disequilibria) and the existence of hybrid individuals show that these exchanges are an actual process rather than a consequence of 'fossil' gene flow. Furthermore, the genetic introgression seems to be asymmetrical in the two regions studied where they come into contact.     

Ceratomyxa aegyptiaca n. sp. (Myxozoa: Myxosporea) from the gall-bladder of Solea aegyptiaca Chabanaud (Pleuronectiformes: Soleidae) in a Tunisian coastal lagoon

A new marine myxosporean species, Ceratomyxa aegyptiaca n. sp. is described from the gall-bladder of Solea aegyptiaca Chabanaud collected from the Ghar El Melh Lagoon in northeastern Tunisia. Mature spores are elongate and crescent-shaped, measuring 8-11?μm in length and 48-58?μm in width. The polar capsules are spherical, 3.2-4?μm in diameter and equal in size. Trophozoites are polysporous and float free in the bile or are attached on the epithelium of the gall-bladder. Morphological data and molecular analysis based on 18S rDNA sequences are provided. The 18S rDNA of C. aegyptiaca is readily distinguishable from that of other myxozoan species, as the genetically most similar myxozoan parasite, C. seriolae Yokoyama & Fukuda, 2001 (AB530265) collected from Seriola quinqueradiata Temminck & Schlegel in Japanese waters, shares with it only 67.5% identical nucleotides over a 1,680-bp long fragment of 18S rDNA.     

Effects of salinity levels on proteome of Suaeda aegyptiaca leaves

Saline soils are the major problem of cultivated lands of Iran. Suaeda aegyptiaca is a salt-tolerant plant (halophytes) that grow naturally in salt-affected areas of Iran. We have employed proteomics to identify the mechanisms of salt responsiveness in leaves of S. aegyptiaca grown under different salt concentrations. Ten-day-old plants were treated with 0, 150, 300, 450, and 600 mM NaCl. After 30 days of treatment, leaf samples were collected and analyzed using 2-D-PAGE. Out of 700 protein spots reproducible detected within replications, 102 spots showed significant response to salt treatment compared to 0 mM NaCl. We analyzed expression pattern of salt-responsive proteins using a hierarchical and two nonhierarchical (Fuzzy ART and SOM) statistical methods and concluded that Fuzzy ART is the superior method. Forty proteins of 12 different expression groups were analyzed using LC/MS/MS. Of these, 27 protein spots were identified including proteins involved in oxidative stress tolerance, glycinebetain synthesis, cytoskeleton remodeling, photosynthesis, ATP production, protein degradation, cyanide detoxification, and chaperone activities. The expression pattern of these proteins and their possible roles in the adaptation of S. aegyptiaca to salinity is discussed.     

Identification of lysine residue at or near active site of luffin-a, a ribosome-inactivating protein from seeds of Luffa cylindrica

Luffin-a, a ribosome-inactivating protein from the seeds of sponge gourd (Luffa cylindrica), was modified with 2,4,6-trinitrobenzenesulfonic acid (TNBS) at pH 8.0 and 20 degrees C. The inhibitory activity of the modified luffin-a on protein synthesis using rabbit reticulocyte lysate was lost rapidly at a rate compatible with that of the modification of a single highly reactive amino group in the initial stage of the reaction. By cation-exchange FPLC of the products of 5-min modification, TNP-luffin-a containing one modified amino group was obtained and shown to have only 6.7% of the activity of native luffin-a without any gross conformational change. The amino acid composition and sequence of the TNP-peptide, isolated by reverse-phase HPLC of the tryptic digest of the TNP-luffin-a, unambiguously demonstrate the trinitrophenylation of lysine residue at position 231. From these results, it was concluded that Lys231 of luffin-a is highly reactive to TNBS and is located at or near the active site of luffin-a.     

Expanding hybrid zone between Solea aegyptiaca and Solea senegalensis: genetic evidence over two decades

Several marine hybrid zones have been described and studied during the last years. Assessing the movements of extending hybrid zones is central to improve our understanding of evolutionary processes. We have re-examined the hybrid zone between Solea aegyptiaca and Solea senegalensis that was first described 22 years ago in northern Tunisia when introgressed S. senegalensis individuals were found in the Gulf of Tunis, whereas locally caught S. aegyptiaca were genetically pure. Six population samples harvested both inside and outside the area where the two fish species coexist were genotyped for allozymes and exon-primed intron length polymorphism. Both types of markers were congruent and revealed that introgression takes place indeed in both directions. A high introgression level (36.4%) in the Bizerta lagoon and much less outside indicate that this is the main area where hybridization occurs while introgression clines towards the south in S. aegyptiaca and towards the north in S. senegalensis plead in favour of the existence of a unimodal hybrid zone. The higher introgression level calculated in the current study (when compared to 16% reported formerly) and the newly found introgressed S. aegyptiaca in Bizerta lagoon seem to indicate that the genetic exchanges occurring between the two taxa are evolving and not stabilized yet.     

Freezing Behavior of Intra- and Extracellular Water of Cultured Green Cells of Lavender as Measured by NMR

Three new proteins which inhibit protein synthesis in rabbit reticulocyte lysates were isolated from an extract of sponge gourd (Luffa cylindricd) seeds by chromatography on a AF-Blue Toyopearl column followed by FPLC with a Mono S column. These three protein-synthesis inhibitory proteins (PSIs) have molecular masses of 19kDa, 15kDa, and 9kDa, and were designated 19K-PSI, 15K-PSI, and 9K-PSI, respectively. Although the 19K-PSI had no effect on protein synthesis in HeLa cells, its inhibitory activity on the cell-free protein synthesis was 340- and 83-fold stronger than those of ricin A-chain and luffin-a, respectively, probably due to hydrolyzing mRNA. The inhibitory activities of 15K-and 9K-PSIs on the cell-free protein synthesis were weaker than those of ricin A-chain and luffin-a. The 19K-PSI was a glycoprotein having an ordinary amino acid composition, three intramolecular disulfide bonds and a blocked N-terminal residue, while the 15K-PSI was extraordinarily rich in glycine and the 9K-PSI in arginine and glutamic acid (and/or glutamine). The amino acid composition of 19K-PSI was: Ser₂₇Glx₃Gly₁₆₄Tyr₇Lys₉His₆, and that of 9K-PSI was: Asx₃Glx₂₅Pro₂Gly₅Lys₂His₂Arg₂₅Trp₃.     

丝瓜的三聚体G蛋白的初步研究

根据拟南芥（ａｒａｂｉｄｏｐｓｉｓｔｈａｈｌｉａｎａ）ＧＰＡ１的保守区段Ａ设计一对特异引物（５′ｃｔｇｇｇｇａａｔｃｔｇｇａａａａｔｃ３′,５′ｃａｃａｇｃｔｇｔａｃａｃｃｔｃａａａｃ３′）通过ＰＣＲ从丝瓜核基因组中扩增植物的三聚体Ｇ蛋白α亚基编码基因,获得了２个片段（ＬＦＧ１,ＬＦＧ２）,并已克隆和测序（已在ＥＭＢＬ数据库中登记,登记号为：ｙ１５２７０,ｙ１５２７１）．序列分析表明ＬＦＧ１和ＬＦＧ２分别由１５１５ｂｐ和７３２ｂｐ构成,都含有三聚体Ｇ蛋白α亚基编码基因的保守区段Ａ,但也都含有内含子．根据片段的大小及ＰＣＲ的特性,ＬＦＧ１可能是丝瓜三聚体Ｇ蛋白α亚基编码基因上的片段．     

Phelipanche aegyptiaca parasitism impairs salinity tolerance in young leaves of tomato

The parasite Phelipanche aegyptiaca infests tomato, a crop plant that is commonly cultivated in semi‐arid environments, where tomato may be subject to salt stress. Since the relationship between the two stresses —salinity and parasitism – has been poorly investigated in tomato, the effects of P. aegyptiaca parasitism on tomato growing under moderate salinity were examined. Tomatoes were grown with regular or saline water irrigation (3 and 45 mM Cl^?, respectively) in soils infested with P. aegyptiaca . The infested plants accumulated higher levels of sodium and chloride ions in the roots, shoots and leaves (old and young) under both salinity levels vs. non‐infected plants. There was a positive linear correlation between P. aegyptiaca biomass and salt accumulation in young tomato leaves, and a negative linear correlation between parasite biomass and the osmotic potential of young tomato leaves. Concentrations of the osmoprotectants proline, myoinositol and sucrose were reduced in infected tomato plants, which impaired the host's osmotic adjustment ability. The sensitivity of P. aegyptiaca to salt stress was manifested as a decrease in biomass. In conclusion, P. aegyptiaca parasitism reduced the salt tolerance of tomato plants by promoting the accumulation of salts from the rhizosphere and impairing the host's osmotic adjustment ability.     

Isolation of a ribosome-inactivating and abortifacient protein from seeds of Luffa acutangula

A glycoprotein with a molecular weight of 28,000 as estimated by SDS-polyacrylamide gel electrophoresis was isolated from seeds of Luffa acutangula using a procedure that involved acetone precipitation, ion exchange chromatography on CM Sepharose CL-6B and gel filtration on Sephadex G-50. In immunodiffusion studies it was found to be immunologically distinct from abortifacient proteins isolated from other members of the Cucurbitaceae family including Momordica charantia, Momordica cochinchinensis, Trichosanthes kirilowii and Trichosanthes cucumeroides. There were some differences in amino acid composition among the proteins although there was a gross similarity. The protein from L. acutangula was capable of inducing mid-term abortion in mice and inhibiting protein synthesis in a cell-free system.     

Purification,characterization and chemical modification studies on a translation inhibitor protein from Luffa cylindrica

A ribosome-inactivating protein (RIP), luffin has been isolated from the seeds of Luffa cylindrica of Cucurbitaceae family by ammonium sulfate fractionation followed by cation exchange and gel-filtration chromatography. Extensive physico-chemical, immunological and biological characterizations were carried out on luffin and compared with that of gelonin. The molecular mass of luffin was -28 kDa as determined by gel-filtration chromatography and SDS-PAGE. The epsilon-NH2 group(s) of luffin were sequentially modified by N-succinimidyl 6-[3-(2-pyridyldithio) propionamido] hexanoate (LC-SPDP), N-succinimidyl-3-(2-pyridylthio)propionate (SPDP) and 2-iminothiolane (2IT) and their effect on immunoreactivity and ribosome inactivating property was evaluated. Modification of single amino group resulted in about 80% inhibition of immunoreactivity and more than 90% loss of protein synthesis inhibition activity. Modification of 2-3 amino groups further hampered both immunoreactivity and protein-synthesis inhibition property LC-SPDP modification played more pronounced effects on immunoreactivity and RIP activity than that of SPDP. However, 2IT modification retained both the immunoreactivity and RIP activity of luffin-LC-SPDP substantially. SPDP showed more pronounced effect on immunoreactivity and RIP activity as compared to 2IT. Therefore, it seems that the positive charge on lysine residues plays an important role in immunological as well as protein synthesis inhibitory effect of luffin.