Mitotic cycle time and somatic selection after neutron irradiation of germinating tomato seeds

Mitotic cycle time was estimated by means of a natural method in control and irradiated material of a dwarf tomato line in order to express the intensity of somatic selection in terms of this biological factor. In the natural method the mitotic index is estimated in relation to the number of hours after germination. The time interval between successive peaks in the mitotic index is considered to be an indication of mitotic cycle time. In control material mitotic cycle time was also estimated by the continuous colchicine method and compared with the natural method.The investigations reported here were performed at the Genetics Department, Catholic University, Nijmegen, Director Prof. Dr. S. J. Geerts and the Institute for the Application of Atomic Energy in Agriculture, Wageningen, Director Dr. D. de Zeeuw.     

Degradation of storage proteins in germinating seeds

The criteria for the participation of proteases in the mobilization of storage proteins during seed germination are formulated. The proteases that satisfy these criteria, namely the acid cysteine endopeptidases, serine carboxypeptidases and neutral aminopeptidases, are reviewed. The possibility of other seed proteases participating in storage protein degradation is discussed. The course of 11S and 7S storage protein degradation through the action of endogenous and exogenous proteases is described. The 11S and 7S proteins are modified during the early stages of proteolysis and the effects of these modifications on the regulation of breakdown are examined. A scheme for 11S protein degradation in germinating seeds is presented.     

Effect of irradiation and/or leucocyte filtration on RBC storage lesions

Red blood cell (RBC) storage lesions have been shown to be associated with some adverse reactions; numerous studies have focused on the lesions caused by storage, and few data on the RBC storage lesions caused by prestorage treatments of leucocyte filtration and irradiation. In this study, we examined the changes related with the RBC storage lesions, including 2,3-diphosphatidylglyceric acid (2,3-DPG), pH, free hemoglobin (Hb), supernatant free K+ and Na+ concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH). Along with the increasing storage time, decreases in 2, 3-DPG levels, pH and Na+ concentration, increases in K+ and free Hb concentrations, and significant morphological changes in RBC in all groups were found. The changes in the groups of irradiation, leucocyte filtration and the combined irradiation and leucocyte filtration were more significant than those in the untreated group. Meanwhile, the MCV levels of the three treated groups were significantly lower than those in the untreated group, while the MCH variations were significantly higher. Our results suggest that irradiation and leucocyte filtration before storage may aggravate blood storage lesions.     

Coffee seeds contain 11S storage proteins

A legumin-like seed protein was purified from the endosperm of coffee ( Coffea arabica L. cv. Colombia). In contrast to legumes, where efficient storage globulin extraction requires buffered saline solutions well above the acidic pK_I of the globulins, coffee legumin is readily extracted with acidic aqueous buffers. The coffee legumin migrates like other 11S storage globulins in sucrose gradients. Subunits of coffee legumin have an apparent molecular mass of about 55 kDa after one-dimensional SDS-polyacrylamide gel electrophoresis in the absence of a reducing agent. In the presence of 2-mercaptoethanol, two polypeptides appear that have apparent molecular masses of 33 and 24 kDa. Two full-length cDNAs were generated from mRNA of developing seeds that were more than 98% homologous. They had open reading frames of 1 458 and 1 467 bp. Each encoded legumin precursors of 486 and 489 amino acids, respectively (M_r=54 136 and 54 818). Examination of a 5' promoter region from a coffee legumin gene revealed a putative legumin-box. Genomic DNA from C . arabica was digested with six different restriction endonucleases. After separation of the fragments by electrophoresis, single discrete fragments on DNA blots hybridized strongly to a cDNA probe for the acidic chain. Other fragments that hybridized weakly with this probe were visible after hybridization at very low stringency. DNA from other species and commercially important cultivars that comprise the genus Coffea produced similar results. Immunocytochemical studies revealed that some legumin was detected in the cytoplasm in mature coffee seeds, but that the majority of it was in large storage vacuoles that accounted for most of the cell volume.     

Desiccation and storage studies on Dipterocarpm seeds

The relationship of seed moisture content (fresh weight basis) to germination, and the effect on viability of various storage conditions were examined for five species of the tropical forest tree genus Dipterocarpus. It was shown that seeds fall into two groups with regard to desiccation tolerance. Firstly, D. obtusifolius and D. turbinatus cannot be dried below about 45% moisture content without damage; a sigmoid curve was found to fit the relationship between germination and moisture content for the latter species. Secondly, D. intricatus, D. tuberculatus and D. alatus can be safely dried to 10%, 12% and 17% moisture contents respectively, but desiccation to near 7% moisture content reduced viability by at least a half. Storage studies showed that seed of D. intricatus and D. tuberculatus possessed increased longevity as moisture contents were reduced within the range 6–20%. It was concluded that seeds in the first group are ‘recalcitrant’ and that those in the second group are ‘orthodox’ in their storage physiology, according to the categories described by Roberts (1973). Wide differences between species in seed desiccation rates were observed. In 15% relative humidity D. intricatus dried to 7% moisture content within a week, whilst D. obtusifolius retained 30% moisture content even after 5 wk; other species had intermediate desiccation rates. Seed size and structure may partly account for the differences observed. Correlations were observed between seed storage physiology and other factors which were investigated. ‘Orthodox’ seeds had quicker desiccation rates, were derived from drier habitats, and had smaller embryos than those of ‘recalcitrant’ seeds. ‘Orthodox’ seeds, with the possible exception of D. alatus, should be kept at 0–3°C with about 12% moisture content in the short term and, provided less than 10% germination is lost on freezing, at-18°C with about 8% moisture content in the long term. ‘Recalcitrant’ seeds should be stored in ventilated containers at 21°C and with moisture contents above 45–50%.     

Biosynthesis of storage proteins in developing rice seeds

Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the starchy endosperm protein of rice (Oryza sativa L. Japonica cv Koshihikari) during seed development confirmed that storage protein begins to accumulate about 5 days after flowering. Two polypeptide groups, 22 to 23 and 37 to 39 kilodaltons, the components of glutelin, the major storage protein in rice seed, appeared 5 days after flowering. A 26-kilodalton polypeptide, the globulin component, also appeared 5 days after flowering. Smaller polypeptides (10- to 16-kilodaltons) including prolamin components, appeared about 10 days after flowering. In contrast, the levels of the 76- and 57-kilodalton polypeptides were fairly constant throughout seed development. Transmission electron microscopy and fractionation by sucrose density gradient centrifugation of the starchy endosperms at various stages of development showed that protein body type II, the accumulation site of glutelin and globulin, was formed faster than protein body type I, the accumulation site of prolamin.

The 57-kilodalton polypeptide but not the glutelin subunits was labeled in a 2-hour treatment with [¹⁴C]leucine given between 4 and 12 days after flowering to developing ears. In vivo pulse-chase labeling studies showed the 57-kilodalton polypeptide to be a precursor of the 22 to 23 and 37 to 39 kilodalton subunits. The 57-kilodalton polypeptide was salt-soluble, but the mature glutelin subunits were almost salt insoluble.

In vitro protein synthesis also showed that the mRNAs directly coding the 22 to 23 and 37 to 39 kilodalton components were absent in developing seeds and that the 57-kilodalton polypeptide was the major product. Thus, it was concluded that the two subunits of rice glutelin are formed through post-translational cleavage of the 57-kilodalton polypeptide.

     

种子顽拗性的形成机理及其保存技术

根据种子的脱水行为将种子分为正常性种子、顽拗性种子和中间性种子。顽拗性种子在发育的末期不经历成熟脱水．脱落时有相对高的含水量,并且对低温和脱水干燥非常敏感。在自然界,顽拗性种子存在一个连续群,即低度、中度和高度顽拗性种子,其差异在于对脱水伤害的敏感程度。影响种子顽拗性的因素,既有种子本身的生理生化物质基础,也有种子在母株上发育过程中所经受的外界环境的影响。目前,对种子脱水耐性的分子机制及其保存技术研究得较多。本文综述了有关顽拗性种子研究的近期进展。     

Effect of humoral factors of the spleen after irradiation]

It is stated that therapeutic effect of humoral factors isolated from the cattle spleen is associated with its influence on alpha 2-macroglobulin (alpha 2-MG) performing a protective function. The results obtained permit recommending the isolated preparations to increase total resistivity of the organism under irradiation.     

Effect of conditions and duration of storage on composition of essential oil from coriander seeds]

The composition of volatile components of the essential oil extracted from seeds of coriander (Coriandrum sativum L.) grown in different years in either Russia or Georgia was studied by capillary gas chromatography. Climatic conditions had a weaker effect on the essential oil composition than the region of growth. After one-year storage in the dark, minor changes were observed in the oil composition, and its organoleptic properties were virtually unchanged. However, the essential oil underwent significant chemical transformation of monoterpenes when stored in the light.     

Cryopreservation and low-temperature storage of seeds of Phaius tankervilleae

In this study we established reliable methods for conservation of seeds of Phaius tankervilleae as an orchid genetic resource. The seeds, which were dehydrated to 5% water content and preserved at 4°C, showed no decrease in viability and germinability after three months. After storage for six months, however, the seeds showed a drastic decrease in germinability, even though survival rate was high. For long-term preservation of seeds of P. tankervilleae, cryopreservation is applied to the freshly harvested seeds. When the seeds were cryopreserved by the vitrification method for up to 12 months there was no apparent deterioration effect of storage time. These results indicate that cryopreservation by the vitrification method is useful for long-term conservation of P. tankervilleae seeds, which are difficult to preserve for more than three months under dry and low-temperature conditions.     

Evaluation of reactive oxygen metabolites in frozen serum samples. Effect of storage and repeated thawing

Measuring the free radical activity in serum samples from prospective studies is the best way to investigate the association between oxidative stress and human diseases. Prospective studies require the analysis of serum samples that have often been stored for a long time. Our study was designed to determine the effect of storage at -30 degrees C and -80 degrees C for two years on free radical activity. We analyzed the free radical activity by measuring circulating hydroperoxides in a pool of sera at baseline and after one day, one week, one month and 25 months of storage, using a photometric method (d-ROMs test). Measurements were performed in aliquots thawed only once at each time point and in aliquots frozen and thawed repeatedly over the study period. After two years we observed a small but statistically significant 4% decrease in the hydroperoxide concentration, which was substantially unaffected by storage temperatures and repeated freeze-thaw cycles. We also carried out the d-ROMs test in sera from ten apparently healthy volunteers at 2, 8, 24, and 48 hours after collection and storage at 4 degrees C and did not observe any significant variation. In conclusion, the d-ROMs test is a simple method suitable to evaluate the free radical activity in frozen serum samples after long-term storage.     

Oxygenation of unsaturated fatty acids in seeds during storage

Seeds of Cichorium intybus L., Crepis thomsonii Babc, and Crepis vesicaria L, were stored from 4 to 8 years at 5°C and then for 18 months under a variety of conditions. Oxygenated acids in Cichorium intybus oil increased from approximately 1% initially to 3% in the first storage period and to 17% while stored at room temperature during the second period. The corresponding levels at these three stages for Crepis thomsonii were 2, 6 and 18%. By gas chromatography (GC) and GC-mass spectrometry, the major oxygenated acids formed during storage were identified as hydroxy acids with conjugated unsaturation and 9,10-epoxy acids. In Crepis vesicaria seed, oil of which contained 53% vernolic (12,13-epoxy-9-octadecenoic) acid originally, approximately 2% of 9,10-epoxides were formed during the storage at room temperature. Levels of hydroxy acids with conjugated unsaturation in this species were 0.3% initially, 2% after 5 years at 5°C, and 9% after 18 months at room temperature. Primary substrates from which oxygenated acids were formed in the three species were crepenynic and linoleic acids, and the almost exclusive formation of 9,10-epoxide from linoleic acid indicated enzymatic involvement.     

Effect of some spices as preservatives for storage of lentil (Lens esculenta L.) seeds

The high antimycotic activity of four kinds of spices,viz. Chinese cassia, cinnamon, clove and thyme, were tested as preservatives for agricultural commodities. Clove, thyme and a mixture of the four kinds of spice completely inhibited aflatoxin production on lentil seeds during eight weeks of incubation. Chinese cassia and cinnamon partially inhibited the aflatoxin production.     

Legumin and vicilin,storage proteins of legume seeds

The structure, location in the seed and distribution of the storage protein of legume seeds are described. Methods which have been employed for the extraction, purification and characterisation of seed globulins are reviewed in relation to modern biochemical practice. The physical, chemical and immunological characteristics of the classical legumin and vicilin preparations from Pisum sativum are summarised and the distributions of proteins with sedimentation coefficients and/or immunological determinants similar to those of legumin and vicilin, are tabulated. The structure and composition of various purified legumin and vicilin-type proteins from a variety of legumes, are compared.     

Inverse correlation between growth and degrading enzyme activity of seedlings after gamma and neutron irradiation of pea seeds

The effect of gamma-, 14 MeV neutron- and fission neutron irradiation was investigated on the growth rate and degrading enzyme activities of pea seedlings. Both dormant pea seeds and 4-day-old growing seedlings were used for the experiments. Depending on the gamma dose between 15 and 300 Gy the height of pea seedlings was found shorter, and parallel with this the endogenous RNase and peroxidase activities were higher than in the unirradiated controls. Seedlings proved to be more sensitive by about one order of magnitude than seeds. Irradiation of seeds between 5 and 10 Gy slightly enhanced the growth rate of seedlings (10 per cent) and parallel with this, the RNase activity measured was lower than that in the controls. On irradiation of seedlings with 14 MeV neutrons the growth inhibition and RNase activity enhancement was only 1.3 times more effective than in the case of irradiation of seeds. The following RBE values were obtained after irradiation of seeds, related to the biological effect of gamma rays: in growth inhibition, 6 for 14 MeV neutrons and 12 for fission neutrons, and the enhancement of two enzyme activities was 15-30 for 14 MeV neutrons and 45-58 for fission neutrons. In the case of seedling irradiation with 14 MeV neutrons the RBE was 1.0 for growth inhibition and between 3 and 6 for enhancement of enzyme activity. The isoenzyme pattern of RNase also changed: two isoenzymes became predominant after the gamma irradiation of seeds, characterized by molecular weights of 21,000 and 30,000, respectively. As a result of enhanced RNase activity, the degradation of longer polysomes to monomeric ribosomes occurred. Thus after ionizing irradiation of pea seeds and seedlings an inverse correlation was found between the growth rate of pea seedlings and the activities of degrading enzymes.