Stimulation of Phospholipid Biosynthesis during Frost Hardening of Winter Wheat

Lipids were labeled with ³³P during frost hardening of two varieties of winter wheat (Triticum aestivum), hardy Kharkov and much less hardy Champlein. The main labeled compounds were phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylglycerol. With time of incorporation the proportion of the radioactivity incorporated into the lipids increased in phosphatidylcholine, especially in Kharkov and at 1 C. During hardening, phospholipid synthesis was greatly stimulated in Kharkov, but much less in Champlein. The proportion of the phospholipids synthesized changed only little with hardening, with a trend towards an increase in phosphatidylcholine. Increased phospholipid synthesis does not seem to be a prerequisite to hardening in winter wheat. However, a high rate of phospholipid synthesis may be required to maintain frost resistance.     

Changes in Amino Acid Composition during Germination of Soybean

Malonogalactan, a malonylated polysaccharide (—74° (c=1.6, H₂O)) produced by Penicillium citrinum, consisted of d-galactose and malonic acid in the approximate molar ratio of 3:1. Molecular weight of the demalonylated galactan (-99° (c=4.6, H₂O)) was about 40,000. From the data regarding optical rotation, nuclear magnetic resonance spectrum, infrared spectrum, glycosidase susceptibility, periodate oxidation, Smith degradation, methylation and acid hydrolysis, the possible structure of the Penicillium malonogalactan is deduced as follows: A galactan, 1,5-β-galactofuranoside polymer esterified with malonic acid at the position of 2 or 3.     

Changes in Amino Acid Composition during Germination of Soybean

α- and γ-Glutamylaspartic acids were detected in acidic fraction of soybean seedling. R_F values of both peptides were consistent with those of authentic samples with several solvent systems.     

Changes in Amino Acid Composition during Germination of Soybean

As a fundamental approach to the problem of amino acid metabolism in soybean, changes in content of twenty-three free amino acids, two acidic peptides, ammonia, ethanolamine, urea and seventeen total amino acids of cotyledon, hypocotyl and root of soybean during germination were determined with an amino acid analyzer. Glycine Max M. var. T201 (non-nodule-forming) and Glycine Max M. var. T202 (nodule-forming) were used for this experiment. The content and composition of free and total amino acids of cotyledon in both T201 and T202 differ from those of other tissues in any stage of germination. However, no significant difference between these two varieties of soybean has been recognized in patterns of free and total amino acids changes during germination.

In dry bean and initial stage of germination a relatively large unknown peak appeared and disappeared thereafter when the change in free amino acid content during germination of soybean was analyzed with amino acid analyzer. From various tests on the unknown peak, it became obvious that the peak was consisted of two peptides, γ-glutamyl-tyrosine and γ-glutamylphenylalanine, which were discovered in soybean by Thompson et al. in 1962. The content of these peptides did not change during the first 20 hours of germination, but they decreased rapidly thereafter and disappeared after 70 hours.     

Changes in Lipid Composition and Carbohydrate Composition of Aspergillus niger Conidia during Germination

Data on the lipid composition and carbohydrate composition of Aspergillus niger conidia make it possible to characterize the individual germination stages and differentiate between the conidia capable of germination and those that lost the germination capacity. The following criteria are proposed: the ratio of phosphatidylcholine and phosphatidylethanolamine, the ratio of mannitol and arabitol, and the levels of sterols and free fatty acids. The role of these compounds in the biochemical background of cell transition from dormancy to active metabolism and their use as indices of the quality of inocula in biotechnological processes are discussed.     

Seasonal Changes in the Composition and Content of Dehydrins in Winter Wheat Plants

Seasonal changes in the pattern and content of dehydrins in winter wheat (Triticum aestivum) plants grown under field and laboratory conditions were studied by one-dimensional PAGE and immunochemical methods. During hardening, plants accumulated dehydrin-like polypeptides with mol wts of 209, 196, 66, 50, and 41 kD. In winter, low-molecular-weight dehydrins with mol wts of 24, 22, 17, 15, and 12 kD were synthesized and accumulated as well. Their content dropped sharply in spring when plants became unhardened. Accumulation/disappearance of these proteins corresponded to the fluctuations in wintering plant frost tolerance before winter and in spring. It is assumed that both high- and medium-molecular-weight dehydrins are involved in plant stress responses and adaptation, whereas low-molecular-weight dehydrins are evidently involved only in the process of low-temperature adaptation.     

A Comparison of Changes in the Germination Responses and Phospholipid Composition of Naturally and Artificially Aged Tomato Seeds

The changes in germinability and phospholipid content of naturallyand artificially aged tomato seeds, variety Kingley Cross, werecompared. Subsequent low temperature pre-sowing treatment (LTPST)was unable to prevent decreases in the germination capacityof aged samples, but significantly improved germination rates.However, in contrast to previous studies on artificial ageingusing the variety Moneymaker, losses of total phospholipid werenot always well correlated with decreased germinability. Totalphospholipid levels in naturally aged seeds were largely unaffectedby pre-sowing treatment, but the proportion of phosphatidylcholinewas greatly increased. These results strongly suggest that lossesof phospholipid are coincidental rather than causative eventsin the loss of seed viability during ageing, and that the patternof changes depends on the type of ageing experienced by theseeds. Controlled deterioration, seed ageing, tomato seeds, seed treatment, phospholipids     

Changes of Template Activity and Proteins of Chromatin during Wheat Germination

Relationships between changes in template activity and composition of chromatin during germination of wheat embyros (Triticum aestivum L.) were investigated. The template activity of chromatin was determined with exogenous DNA-dependent RNA polymerase II (EC 2.7.7.6) prepared from wheat embryos. It was essentially constant for 18 hours of germination, corresponding to 2.5% of that of a native calf thymus DNA. Thereafter, the activity increased 2-fold and 5-fold at 24 and 60 hours of germination, respectively.     

Changes in Level and Activity of Phospholipid Transfer Protein during Maturation and Germination of Maize Seeds

The variations of the amounts of phospholipid transfer proteins (PLTP), determined by ELISA and immunoblotting methods, were followed during the maturation and germination of maize (Zea mays L.) seeds. Changes of the amounts of PLTP occur during seed maturation. The levels of PLTP, low in the first 3 weeks after fecondation, strongly raised 3 to 5 weeks after, then reached and maintained a high value (10% of total soluble proteins) during the last steps of maturation. These variations, determined by ELISA, are in accordance with the observations made by immunoblotting. Changes in phospholipid transfer activity were also found when protein extracts prepared from seeds at different stages of maturation were assayed for transfer activity. The levels of PLTP were also determined during the germination of maize seeds and the early growth of the plantlets, both in the endosperm and the aerial parts. While no major change was observed in the endosperm, a high increase in PLTP level was found in the aerial part of the plantlet, both by ELISA and immunoblotting. An enhancement of the phospholipid transfer activity was parallely observed in the protein extracts of plantlets at various stages of germination. These results are consistent with an in vivo correlation between the synthesis of phospholipid transfer protein, observed during the maturation and germination of maize seeds, and the biogenesis of membranes which involves intracellular movements of phospholipids.     

Changes in the Activities of Pyrrolooxygenases during the Germination of Wheat Grains

Porphobilinogen oxygenase, skatole pyrrolooxygenase, and tryptophan pyrrolooxygenase were found in the different parts of germinating wheat (Triticum aestivum) grain seedlings. In the embryos of grains germinated for 24 hours, the activities of PBG oxygenase and skatole pyrrolooxygenase were inhibited by a labile inhibitor. Tryptophan pyrrolooxygenase activity was not inhibited. Embryos of grains germinated for 48 hours showed higher activities for the three enzymes. The latter were also present in the radicles and coleoptiles of 96-hour germinated wheat grains. A DEAE-cellulose analysis of a crude enzymic preparation from embryos allowed the separation of two molecular forms of the three pyrrolooxygenases. The more cationic forms of porphobilinogen oxygenase and skatole pyrrolooxygenase were associated with the inhibitor. This form of porphobilinogen oxygenase had allosteric kinetics while the more anionic form had Michaelis kinetics. Both forms of skatole pyrrolooxygenase had Michaelis kinetics. The activity of tryptophan pyrrolooxygenase was highest in seedling roots and was found to be inhibited in seedling young leaves. This enzyme oxidized tryptophanyl dipeptides, as well as a nonapeptide, to N-formylkynurenine-containing peptides. The pyrrolooxygenase also oxidized the tryptophanyl residues of lysozyme, chymotrypsin, and trypsin.     

Changes in Ultrastructure and Subunit Composition of Protein Body in Rice Endosperm during Germination

Changes in ultrastructure of protein bodies in subaleurone cells of rice endosperm during germination were studied by transmission electron microscopy. The subaleurone cells contained two different types of protein bodies: PB-I (spherical) and PB-II (crystalline). Both types of protein bodies were deconstructed during germination. But there was a considerable difference in digestibility between PB-I and PB-II. PB-II which did not have a dense core was easily digested from the central portion when germination began. At 6 days of germination, PB-II was almost deconstructed. On the other hand, PB-I which displayed concentric rings with a dense core was digested from the outside after 3 days of germination. At 9 days of germination, many kernels of the spherical protein bodies remained.

Changes in subunit composition of protein bodies during germination were investigated by SDS-polyacrylamide gel electrophoresis. Protein body fractions were isolated from germinating grains at various stages by enzymatic digestion and two-phase system, then subjected to SDS-polyacrylamide gel. As germination proceeded, 15 (b₁), 20 (d₁), 24 (e), 35 (f₁) and 37 (f₃) kdaltons subunits decreased. On the other hand, 16 (b₂), 21 (d₂) and 36 (f₂) k daltons subunits remained at the later stage of germination. We think that PB-I contains b₂, d₂ and f₂ subunits and is attacked only from the outside at middle and later stages of germination by de novo protease. On the contrary, PB-II contains b₁ d₁ e, f₁ and f₃ subunits is utilized at an early stage of germination. PB-II may possibly contain latent protease. The breakdown process of PB-I was by exo-type digestion, on the contrary, that of PB-II was by endo-type digestion.     

干旱条件下冬小麦幼苗根细胞膜脂组成的变化

干旱条件下小麦幼苗根膜脂总脂肪酸含量、磷脂含量及总脂肪酸双键指数均下降,而游离甾醇含量却明显增加,结果导致游离甾醇／磷脂比率上升。用薄层层析法测得小麦根细胞磷脂主要由磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)及磷脂酸(PA)组成。干旱降低了各种磷脂的含量,但不改变其相对配比。文中讨论了膜脂代谢变化与植物抗旱性的关系。     

小麦品种冀麦24抗麦红吸浆虫QTL分析

麦红吸浆虫是影响小麦产量和品质的重要害虫,研究小麦对吸浆虫抗性的遗传及其连锁分子标记对于提高抗虫品种的选择效率具有重要意义。本研究以小麦感虫品系6218与抗虫品种冀麦24产生的重组近交系(RIL)群体为材料,利用SSR标记和人工虫圃对冀麦24的抗虫性遗传进行了研究。结果表明:6218与冀麦24的抗性差异显著,RIL群体在2年2点的鉴定中抗性稳定;所构建的遗传连锁图谱包含112个SSR位点,形成26个连锁群,图谱全长835.7 cM,标记间平均距离为7.5 cM。利用QTL IciMapping的完备区间作图法,在4A染色体上检测到1个加性效应位点(QSm.hbau-4A),该位点在2个鉴定年度的贡献率分别为9.67%、10.57%。该抗性QTL及其连锁SSR标记的发掘,将有助于提高小麦抗吸浆虫育种的选择效率。     

Metabolic and Ultrastructural Changes in Winter Wheat during Ice Encasement Under Field Conditions

The effect of ice encasement on the physiological, metabolic, and ultrastructural properties of winter wheat (Triticum aestivum L.) grown under field conditions was examined by artificially encasing winter wheat in ice during early winter. Cold hardiness and survival of ice-encased seedlings declined less rapidly in Kharkov, a cold-hardy cultivar than in Fredrick, a less hardy cultivar. Ethanol did not accumulate in non-iced seedlings, but increased rapidly upon application of an ice sheet. Lactic acid accumulated in both cultivars during late autumn, prior to ice encasement, and elevated levels of lactic acid were maintained throughout the winter in seedlings from both iced and non-iced plots. The rate of O₂ consumption of shoot tissue of seedlings from non-iced plots remained relatively constant throughout the winter, but declined rapidly in seedlings from ice encased plots. Major ultrastructural changes did not occur in shoot apex cells of non-iced winter wheat seedlings during cold hardening under field conditions. However, the imposition of an ice cover in early January resulted in a proliferation of the endoplasmic reticulum membrane system of the cells, frequently resulting in the formation of concentric whorls of membranes, often enclosing cytoplasmic organelles. Electrondense areas within the cytoplasm which appeared to be associated with the expanded endoplasmic reticulum were also frequently observed.     

The Phospholipid Composition of Nuclear Subfractions from Germinating Wheat Embryos

The content of phospholipids in chromatin, nuclear matrix, and nuclear membrane from wheat (Triticum aestivum L.) embryos was studied. Subfractions of intact nuclei from dry embryos were shown to differ in the content and composition of particular phospholipids. Embryo germination resulted in the redistribution of phospholipid between nuclear subfractions. A functional role of structural changes in the nuclear membrane due to this phospholipid redistribution is discussed. It is supposed that these rearrangements change nuclear membrane permeability and its surface charge.     

Developmentally Regulated Changes in Phospholipid Composition in Murine Molar Tooth

In order to explore the possibility that phospholipids are differently expressed during the cascade of events leading to tooth formation, we decided to carry out simultaneous biochemical, histological and electron histochemical studies. High performance thin-layer chromatography and gas–liquid chromatography were used to compare the composition of embryonic mouse first molar tooth germs at day 18 of gestation (E18) and at birth (D1), erupting teeth at day 7 (D7) and erupted molars at day 21 (D21). For the latter, non-demineralized and EDTA-demineralized lipid extracts were analysed separately. Moreover, an ultrahistochemical study was carried out using the iodoplatinate reaction which retains and visualizes phospholipids. Developmentally regulated changes occurred and were closely correlated with an increase in cell membrane phospholipids. Gradual accumulation of phospholipids was identified in the extracellular matrix, at an early stage of tooth germ development within the basement membrane and later, as predentine/dentine and enamel components participating in mineralization processes. Matrix vesicles transiently present in dentine were partly responsible for the lipids that were detected. A first group of phospholipids including phosphatidylcholine as the major membrane-associated phospholipid and phosphatidylinositol as the intracellular second messenger increased by a factor of 2.3 between E18 and D21. This increase is probably associated with cell lengthening and was relatively modest compared with the higher increase detected for a second group of phospholipids, namely phosphatidylethanolamine (×4.8), phosphatidylserine (×5.9) and sphingomyelin (×5.4). This second group of extracellular matrix-associated phospholipids constituted 68% of the demineralized lipid extract and, therefore, contributes to the mineralization of dental tissues.     

冬小麦种子萌发过程中的结合态玉米赤霉烯酮

冬小麦种子萌发过程中的结合态玉米赤霉烯酮陈新建（河南农业大学农学系,郑州１５０００２）孟繁静（北京农业大学生物学院,北京１０００９４）关键词结合态玉米赤霉烯酮,冬小麦玉米赤霉烯酮（ｚｅａｒａｌｅｎｏｎｅ）是玉米赤霉菌（Ｇ～ｔｈｅ－）的一种次生代谢产物...     

Ultrastructural Changes during Swelling and Contraction of Mitochondria from Cold-hardened and Non-hardened Winter Wheat

Mitochondria isolated from both 2 and 24 C grown winter wheat (Triticum aestivum L.) undergo spontaneous swelling in isomolar KCI solutions, but only 24 C mitochondria exhibit a substrate-induced contraction response. Electron microscopic examination revealed that 24 C mitochondria have more clearly defined cristae, less matrix material, and are generally more electron-dense than 2 C mitochondria. During swelling, the matrix material of both 2 and 24 C mitochondria expands and the mitochondria become less electron-dense. After partial swelling, 24 C mitochondria contract upon addition of succinate, and regain structural characteristics similar to those of untreated mitochondria. In contrast, mitochondria from 2 C seedlings continue to swell after addition of substrate, and many of the mitochondria become irregular in shape and lose much of their matrix material. A comparison of results obtained from absorbancy measurements, electron microscopy, and a Coulter Counter indicate that swelling and contraction involve changes both in over-all volume, and internal structural characteristics of mitochondria from 2 and 24 C grown seedlings. Electron microscopic examination of shoot cells showed that mitochondria in 24 C grown seedlings possessed more recognizable cristae and greater internal organization than mitochondria in 2 C seedlings.     

Changes in Nicotinic Acid Content and Its Nucleotide Derivatives of Rice and Wheat Seeds during Germination

Relative distribution of bound and free forms of nicotinic acid in rice and wheat seeds and their metabolism during germination were the subject of the present investigation. Measurement of the levels of NAD (nicotinamide adenine dinucleotide) and NADP (nicotinamide adenine dinucleotide phosphate) formed another part of the work. Total nicotinic acid in both rice and wheat increased with germination and was maximum at 72 hours. From this time onwards, it began to decline rapidly and at the end of experiment, i.e., after 120 hours, it was lower than that for ungerminated seeds on per seedling basis, although it was slightly higher on per g dry weight basis. Ungerminated seeds of wheat and rice contained about 89 per cent and 80 per cent respectively of their total nicotinic acid in bound form which became partially free in course of germination. Total nucleotides (oxidised plus reduced forms) increased progressively up to 96 hours followed by a slight fall at 120 hours. NAD reached a maximum at 24 hours and fell gradually thereafter. The depletion of NAD was associated with a progressive accumulation of NADH. NADP decreased from the peak value at 72 hours. Formation of NADP and its maintenance at high level depend on NAD in the oxidised form and the content slowed down in tissues with higher NADU/NAD ratio. A relatively large amount of NADPH was present throughout the experiment.