Content of free amino acids and their exosmose from maize kernels in relation to cold resistance

Maize caryopses were exposed in moist, sterile sand to temperatures of 24°, 10° and 6°. The amino acid content of embryo, endosperm and pericarp tissues of kernels and amino acid occurence in sand eluates was analysed by paper chromatography. The lower the temperature to which the seeds were exposed and the greater the delay in start of germination, the greater were the amounts of amino acids exosmosed from the kernel to the surrounding media. The exudation of aspartic acid, proline, alanine, γ-aminobutyric and glutamic acids was especially high. The time-course of amino acid content in various kernel parts proves that cold does not check the enzymic hydrolysis of reserve nitrogen compounds. There is, however, an inhibition of translocation and use of amino acids in the embryo. Therefore, amino acids diffuse out from the caryopse into the media. Although mainly the peripheral parts of endosperm take part in exosmose, participation of the embryo in the exudation of certain amino acids (proline) can be supposed as well. In cold resistance of emerging maize kernels the remarkable interaction of microorganisms is causally related to the high amino acid exosmose found in these experiments. The mechanism of amino acid exosmose and its relationship to cold resistance are discussed in detail.     

Factors influencing seed dormancy and germination in sand,and seedling survival under desiccation,of Psammochloa villosa (Poaceae), inhabiting the moving sand dunes of Ordos,China

Psammochloa villosa (Trin.) Bor. (Poaceae), is distributed primarily in moving sand dunes of the Ordos Plateau, China. Freshly harvested caryopses (seeds) are in non-deep physiological dormancy (non-deep PD). Germination is slow and low and only over a narrow temperature range. A treatment of four weeks cold stratification at 3 to 5 °C in darkness was required to break non-deep PD, allowing germination to reach high percentages at higher temperatures and without light requirement. Rate and percentages of germination were increased by scarifying the caryopsis coat and by artificial removal of different proportions of the endosperm. However, seedling dry weight and increases in root and shoot lengths, were significantly influenced by the proportion of the endosperm that remained on the caryopses. Higher percentages of seedling emergence were obtained from the shallowly buried caryopses, ranging from 0.5–2.0 cm, and the depth of the caryopses in the sand affected the time of germination. The deeper the caryopses were buried, the more that remained ungerminated and in enforced dormancy. The caryopses germinated when the upper sand layer was removed and buried caryopses were at a suitable sand depth for germination, or when the sand was aerated. In natural habitats, germinated seedlings may be wholly exposed to the air by sand erosion and thus be exposed to drought stress. However, young seedlings have the ability to tolerate desiccation and to recover after rehydration. Root length at the `point of no return' is 4 mm.     

Osmoregulation, growth and sucrose accumulation in germinated Trigonella foenum-graecum (fenugreek) seeds treated with polyethylene glycol

Germinated seeds of Trigonella foenum-graecum L. (fenugreek) were grown in water or in polyethylene glycol (PEG) solutions. After endosperm removal, the water relations, growth, dry weight, sucrose and reducing sugar content of the embryo were determined. Under water sstress conditions, water content and osmotic potential (π₀) at saturation, growth and dry weight were lower than in non-stressed controls. The reduction in dry weight indicated a lower uptake of solutes from the endosperm and the decrease in π₀ was not accompanied by an increase in the amount of the accumulated solutes. It is suggested that embryos of stressed fenugreek seeds control osmotic potential by reduction of water uptake and that this results in reduction of growth. Embryos isolated from germinated seeds ("naked" embryos) were grown in water or in PEG solutions, with or without galactose (as an external solute source substituting for the endosperm). The results indicate that a decrease in the external solute did not account for growth reduction under conditions of water stress, and that decreased solute transport to the embryo may be important. The sucrose contents of "naked" embryos and of embryos from whole seeds were higher after PEG treatment, while reducing sugar contents were lower compared to non-stressed controls. The increased sucrose accumulation may be due to decreased sucrose hydrolysis.     

Is The Barley Endosperm a Water Reservoir for the Embryo When Germinating Seeds Are Dried?

The water content of germinating seeds fluctuates in response to water potential changes in the surrounding environment. We tested the hypothesis that the endosperm functions as a water reservoir when imbibed seeds experience drying, and we characterized water uptake and movement within barley (Hordeum vulgare cv. Triumph) caryopses (hereafter referred to as seeds). Water movement into and through germinating barley seeds during imbibition and drying was determined gravimetrically and with the fluorescent dye trisodium 8-hydroxy-1,3,6-pyrenetrisulfonate (PTS). During imbibition, embryo tissues hydrated more rapidly and reached a higher water content (g H20/g dry weight) than did the endosperm, although the endosperm eventually contained nine times as much total water. When barley seeds that had imbibed for 12 h were exposed to moderate (-4 MPa) drying, PTS solution moved from the endosperm into the shoot meristem, radicle, and scutellum, but not vice versa. Radicle emergence and elongation proceeded for up to 8 h. With harsh (-150 MPa) drying, PTS concentrated almost exclusively in the radicle. These data illustrate that the endosperm is at least a temporary water storage compartment external to the embryo itself. We speculate that water supplied by the endosperm may be important in reducing the harmful effects of drying during the critical transition period when a germinating seed changes from a desiccation-tolerant to a desiccation-intolerant organism.     

Effects of partial endosperm removal on embryo dormancy breaking and salt tolerance of <Emphasis Type="Italic">Hordeum spontaneum</Emphasis> seeds

Seed damage is a common phenomenon in nature and in agricultural production. In this experiment, partial endosperm removal from wild barley (Hordeum spontaneum) caryopses, sampled from three ecotypes originated from xeric environments in Israel, was conducted. The aim was to examine seed dormancy and germination states in damaged caryopses and salt tolerance of young seedlings derived from them. Six treatments were made: (1) control seeds with intact caryopses; (2–4) removal of 0.25, 0.5, and 0.75 of the length of intact caryopses; (5) transection at the points, at which the endosperm and embryo meet; and (6) slitting of endosperm opposite the embryo. A significant negative correlation was found between germination percentage (dormancy release) and the relative distance from the dissection point to embryo. Partial removal of the endosperm could accelerate dormancy release. Seedling salt tolerance was assessed by the ratio of root or coleoptile length in a seedling grown in 100 or 200 mM NaCl solution to that of a seedling grown in water. The seedling salt tolerance was positively correlated with the removed portion of the seed endosperm. For each level of endosperm removal, the salt tolerance to 200 mM NaCl of the seedlings derived from the Dead Sea ecotype was higher than those from both the Sede Boker and the Mehola ecotypes. The results suggest that partial damage to seed endosperms in natural conditions may play a role in increasing the phenotypic plasticity of germination and salt tolerance.     

Nonstructural carbohydrates in dormant and afterripened wild oat caryopses

Nonstructural carbohydrates were determined in both embryo and endosperm of dormant (nongerminating) and afterripened (germinating) intact caryopses of wild oat ( Avena fatua L.). No changes in endosperm starch or soluble sugar were observed at the onset of germination (18 h). No changes in glucose, fructose, sucrose or starch within dormant or afterripened embryos correlated with onset of visual germination. In afterripened embryos, depletion of raffinose (18 h), stachyose (18 h) and galactose (24 h) was correlated with germination. In contrast, raffinose-family oligosaccharide levels in dormant embryos remained constant for 7 days following imbibition. Germination of isolated dormant embryos on 88 m M galactose-containing media was accompanied by decreased endogenous levels of raffinose and stachyose. Isolated embryos from dormant caryopses incorporated ¹⁴C from ¹⁴C-fructose into both raffinose and stachyose during 24 h of imbibition. In contrast, no ¹⁴C incorporation into stachyose was observed in embryos from afterripened caryopses. No ¹⁴C incorporation into raffinose was observed at 18 and 24 h. When in vitro activities of α galactosidase were measured, no temporal differences between dormant or afterripened caryopses were detected in either embryo or endosperm tissue. Although the mechanism associated with differences in utilization of raffinose and stachyose is yet unidentified, alterations in raffinose-family oligosaccharide metabolism in the embryo appear to be a unique prerequisite for afterripening-induced germination.     

Some effects of temperature during seed development on carrot (Daucus carota) seed growth and quality

The growth and development of carrot seeds cv. Chantenay Red-cored Royal Chantenay at day/night temperatures of 20/10°C, 25/15°C and 30/20°C and subsequent seed performance were examined in 1984 and 1985. An increase in temperature from 20/10°C to 30/20°C reduced mean weight per seed by 20% in 1984 and by 13% in 1985. There were no effects of temperature on endosperm + embryo weight, or on endosperm cell number but the weight of pericarp decreased with an increase in temperature. Seeds grown at the highest temperature had the largest embryos and the highest nitrogen, DNA and rRNA content; they germinated and emerged earlier, and gave higher percentage seedling emergence than those grown at the lowest temperature. There were no effects of temperature during seed growth on the rate of imbibition of water by seeds during the germination process.     

Physiology of the cold-resistance of maize during Germination. the reaction of maize (Zea mays L.) to low temperature during germination and its cold-resistance

Attempts to elucidate the physiological basis of cold-resistance during germination led to the investigation of the germination of seeds at low temperature, the effect of pathogenic and parasitic microorganisms being excluded. At the various experimental temperatures (4°, 6°, 8°, 10° and 14°C) it was found that seed samples of different varieties of maize respond to conditions close to the temperature minimum for germination in three quantitatively different ways. Firstly, caryopses germinate considerably later than at higher temperatures, secondly, the kernels will not germinate even after a prolonged period, without losing their viability and, thirdly, in some of the caryopses the embryos perish. The lower the temperature used the greater the inhibition of germination and the higher the mortality of the embryos. The increase in mortality with decreasing temperature indicates that the mortality is primarily due to the actual effect of the low temperature. Samples of seeds displayed quantitative difference in the above mentioned response. Only when fungicide-treated seed material was used for the cold-resistance experiments was a significant relationship found to exist between this property and the general germinating capacity or mortality at the same low temperature.     

A germination-specific endo-beta-mannanase gene is expressed in the micropylar endosperm cap of tomato seeds

Endo-beta-mannanase (EC 3.2.1.78) is involved in hydrolysis of the mannan-rich cell walls of the tomato (Lycopersicon esculentum Mill.) endosperm during germination and post-germinative seedling growth. Different electrophoretic isoforms of endo-beta-mannanase are expressed sequentially in different parts of the endosperm, initially in the micropylar endosperm cap covering the radicle tip and subsequently in the remaining lateral endosperm surrounding the rest of the embryo. We have isolated a cDNA from imbibed tomato seeds (LeMAN2) that shares 77% deduced amino acid sequence similarity with a post-germinative tomato mannanase (LeMAN1). When expressed in Escherichia coli, the protein encoded by LeMAN2 cDNA was recognized by anti-mannanase antibody and exhibited endo-beta-mannanase activity, confirming the identity of the gene. LeMAN2 was expressed exclusively in the endosperm cap tissue of tomato seeds prior to radicle emergence, whereas LeMAN1 was expressed only in the lateral endosperm after radicle emergence. LeMAN2 mRNA accumulation and mannanase activity were induced by gibberellin in gibberellin-deficient gib-1 mutant seeds but were not inhibited by abscisic acid in wild-type seeds. Distinct mannanases are involved in germination and post-germinative growth, with LeMAN2 being associated with endosperm cap weakening prior to radicle emergence, whereas LeMAN1 mobilizes galactomannan reserves in the lateral endosperm.     

Relationship of Endo-[beta]-D-Mannanase Activity and Cell Wall Hydrolysis in Tomato Endosperm to Germination Rates

The endosperm tissue enclosing the radicle tip (endosperm cap) governs radicle emergence in tomato (Lycopersicon esculentum Mill.) seeds. Weakening of the endosperm cap has been attributed to hydrolysis of its mannan-rich cell walls by endo-[beta]-D-mannanase. To test this hypothesis, we measured mannanase activity in tomato endosperm caps from seeds allowed to imbibe under conditions of varying germination rates. Over a range of suboptimal temperatures, mannanase activity prior to radicle emergence increased in accordance with accumulated thermal time. Reduced water potential delayed or prevented radicle emergence but enhanced mannanase activity in the endosperm caps. Abscisic acid did not prevent the initial increase in mannanase activity, although radicle emergence was markedly delayed. Sugar composition and percent mannose (Man) content of endosperm cap cell walls did not change prior to radicle emergence under any condition. Man, glucose, and other sugars were released into the incubation solution by endosperm caps isolated from intact seeds during imbibition. Pregerminative release of Man was suppressed and the release of glucose was enhanced when seeds were incubated in osmoticum or abscisic acid; the opposite occurred in the presence of gibberellin. Thus, whereas sugar release patterns were sensitive to environmental and hormonal factors affecting germination, neither assayable endo-[beta]-D-mannanase activity nor changes in cell wall sugar composition of endosperm caps correlated well with tomato seed germination rates under all conditions.     

Studies on the early development of zygotic and synergid embryo and endosperm in polyembryonic rice ApIII

A morphological, cytological and embryological investigation was conducted on the early development of embryo and endosperm in polyembryonic rice (Oryza sativa L.) ApIII. We found that the percentage of single-, twin- and triple-seedlings from mature caryopses was 82.4–85.6%, 11.3–14.6% and 2.0–3.8%, respectively. From twin-seedlings one of the plants and in the triple-seedlings usually two seedlings were very weak having a mortality of about 50–60% and over 95%, respectively, and most of the dead seedlings were haploid. In addition, among 674 young caryopses the percentage of single-, double- and triple-embryo sacs was 77.60%, 18.10% and 4.30%, respectively. The development of both embryo and endosperm was observed in the embryo sacs of 653 young caryopses, while embryo development occurred in absence of endosperm in the embryo sacs of 21 ones. Furthermore, after pollen tube penetration, frequently one, rarely two, additional embryos arise via apogamy from the typical specialized synergid of normal egg apparatus with a collapsed cap-neck region that had a damaged filiform apparatus at the micropylar end. Finally, the formation of a strikingly degenerated endosperm cell region is a specialized functional structure that serves for the active transport of metabolites from endosperm to the developing embryo. Taken together, these results suggest that synergid apogamy seems to be the unique reason of polyembryony, poor growth and high mortality of many seedlings may be due to haploidy, and high genetic stability of polyembryony occurs in rice ApIII.     

利用原子吸收光谱法和电镜能谱仪法分析重金属Pb、Cd在水稻颖果不同部位中的分配

比较研究了传统原子吸收光谱法与电镜能谱仪法分析重金属Pb、Cd在水稻籽粒不同部位中的分配情况,研究结果表明:两种方法均显示,Pb、Cd在水稻颖果中的分配为胚>糊粉层>果皮>近糊粉层处胚乳>颖果中心,其中,胚、糊粉层和果皮Pb、Cd含量差异较小,整个胚乳Pb、Cd含量变化也不大,但胚、糊粉层和果皮Pb、Cd含量显著高于胚乳.扫描能谱仪能够准确反映重金属在植物不同部位分布的差异,结果与原子吸收光谱法无显著差异.扫描能谱仪法的优点在于操作方便快捷,但是只能用于定性,不能定量测定出某一部位重金属的准确含量,因此,不能代替传统的原子吸收光谱法.     

荔枝胚败育过程中内源激素与蛋白质含量的变化

连续3年(1999-2001年)对典型的荔枝焦核品种桂味、糯米糍和大核品种黑叶、怀枝花后10-40d的幼胚和胚乳内源激素、多酚含量及蛋白质动态变化进行研究。结果表明,焦核品种幼胚及胚乳中的IAA、GAs和ABA含量低于大核品种;多酚类物质含量在胚中低于大核品种,胚乳中则高于大核品种;胚和胚乳中的蛋白质含量均低于大核品种。蛋白质电泳结果显示,22．5、28．5和45kD这3类蛋白质在怀枝和黑叶的胚蛋白质代谢过程中表现出较高的稳定性,桂味和糯米糍胚蛋白质中的28．5kD蛋白质也有相似的特性。     

Studies on the Structure,Afterripening and Cytochemistry of Seeds in Eleutherococcus brachypus Harms

Seeds of Eleutherococcus brachypus Harms were flat-kidney-shaped and their seed coats were only composed of one layer of cells. Embryos with abundant protein in their cells were just at the heart-shaped stage and were capped by sacs formed from degenerating endosperm cells when seeds shed from their maternal plants. A large amount of stored protein grains and lipids existed in endosperm cells but no polysaccharide grains were present either in endosperm cells or in embryo cells. Viable seeds were only 9.27% of the total. The plump seeds germinated in the cultivated field after 18～19 months and their germinating rate was 1.67%. Besides, the content of protein decreased gradually and a few polysaccharide grains were stored in embryo cells during the process. The afterripening process of seeds stratified at different temperatures ended after 6 months and the cytochemistry features of the seeds were that the content of protein decreased gradually and numerous polysaccharide grains had been stored in embryo cells at the late heart-shaped embryo stage and retained till the mature embryo stage. The structure, afterripening and cytochemistry of seeds were compared between Eleutherococcus brachypus and Eleutherococcus senticosus. The poor quality of the seeds, longer time of afterripening in a natural state and much lower germination rate of E. brachypus are considered to be important reasons for the endangerment of this species. Somemeasures are suggested for its conservation based on the above facts.     

Factors affecting the dormancy and germination of bleeding heart [Lamprocapnos spectabilis (L.) Fukuhara] seeds

• Information on the optimal conditions to promote the germination of Lamprocapnos spectabilis (L.) Fukuhara seeds is limited; consequently, this study was conducted to establish the requirements to break seed dormancy and promote germination.
• The selected seeds had morphophysiological dormancy and had not begun embryo development. To study the dormancy breaking and embryo development processes, seeds were subjected to constant or changing temperature treatments during moist stratification.
• High temperature and humidity resulted in vigorous embryo growth, with the longest embryos occurring after 1 month of incubation at 20 °C. At 4 °C, the seeds required incubation period of at least 3 months to germinate. Embryo growth and germination were higher with changing high and low temperatures than under a constant temperature, and changing temperatures also considerably changed the endogenous hormone levels, embryo development and germination. Bioactive gibberellin (GA) content was higher in seeds incubated at 20 °C for 1 month, then at 4 °C for 2 months. The content of endogenous abscisic acid in seeds subjected to the same treatment decreased by 97.6% compared with that of the untreated seeds.
• Embryo growth and seed germination require changing high and low temperatures; however, exogenous GA₃ could substitute for high temperatures, as it also causes accelerated germination. In this study, the seeds of L. spectabilis were identified as an intermediate simple type, a sub‐level of morphophysiologically dormant seeds.

     

Viability of onion (Allium cega) seed as influenced by temperature during seed growth

The yield and performance of seeds from crops of winter-hardy, bolting-resistant onion grown at temperatures of 15–16, 18–19 and 22–23°C in 1979, 1980 and 1982 were compared. Yields of seed from crops grown at 22–23°C were lower than those from crops grown at lower temperatures but the seeds ripened between 11 and 32 days earlier. Seeds from crops grown at mean temperatures of above 18°C gave higher percentage germination when imbibed at 30°C than 20°C and they also gave higher percentage seedling emergence than those from crops grown at lower temperatures. Seedlings from seeds produced at mean temperatures above 18°C were heavier than those from seeds of a similar weight but produced at lower temperatures. None of these differences were associated with differences in seed weight, embryo weight or seed dormancy but were positively correlated with differences in seed N-concentration. The differences were also associated with the rate of imbibition of water as high germination, high N-content seeds had a slower rate of imbibition than low germination, low N-content seeds of the same weight.     

Differential expression of two β-amylase genes of rye during seed development

The expression of two β-amylase loci was analysed in the developing seeds of two inbred lines of rye (Secale cereale L.), one of which was a β-amylase deficient mutant. Enzymatic activity and the contents of enzymatic protein and mRNA specific for each of an endosperm-characteristic and ubiquitous β-amylase were determined throughout the course of caryopsis development. Both loci were expressed in the developing normal line caryopses according to different temporal and quantitative patterns. The ubiquitous enzyme-specific locus β-Amy 2 was expressed earlier; both mRNA and enzymatic protein accumulated to a maximum extent at 10 to 15 days after pollination. In contrast, the highest content of mRNA for endosperm β-amylase (encoded by the β-Amy I locus) was found 20 days after pollination, and the corresponding enzymatic protein accumulated throughout seed development. The expression of the β-Amy I locus was 30- to 40-fold higher than that of the β-Amy 2 locus in terms of maximum specific mRNA accumulation. The expression product of only the β-Amy 2 locus was found in the developing mutant line caryopses. The expression pattern of this locus was similar in the developing normal and mutant line seeds in terms of the temporal accumulation of mRNA and enzymatic protein. However, an approximately 4-fold higher level of ubiquitous β-amylase-specific mRNA was found in the mutant than in the normal line caryopses, and the content of ubiquitous β-amylase protein decreased to near zero at seed maturity in the mutant line, but not in the normal line, caryopses. The enzymatic activities of both β-amylases appeared to be regulated at the level of accumulated enzymatic protein.     

Freezing pattern in apple seeds as affected by the temperature of fruit storage

Storage of apple fruits ( Pyrus malus L. cv. Golden Delicious) at different temperatures (0, 12 and 35°C) markedly altered the pattern of water freezing in the seeds. Higher temperatures of storage brought about a shift from the sequential to the discontinuous type of freezing in seeds, the low temperature exotherm (LTE) being much more pronounced than the high temperature one (HTE). The occurrence of LTE was highly correlated with embryo death. Fruit storage at higher temperatures also caused a decrease in the threshold super cooling temperature of seeds and of naked embryos, removed from the seed coat and endosperm. The decrease was less pronounced in naked embryos than in intact seeds. The results presented show that the frost resistance of apple seeds relies mainly on the supercooling ability of the embryos and is increased by the presence of seed coat and endosperm. The broad peak of the LTE indicates that, in contrast with other seeds and many super cooling organs, massive ice nucleation in apple seeds occurs within the embryo tissues and that extra organ freezing seems to be of less importance. Therefore, the increased super cooling ability of apple seeds, isolated from fruits stored at higher temperatures, seems to rely on those seed properties that protect embryo cells against heterogeneous ice nucleation.