Reversal of (±) -Abscisin II Induced Inhibition of Lettuce Seed Germination and Seedling Growth by Kinetin

Observations have been made on the effect of synthetic, (±)-abscisin II (dormin), alone and in combination with gibberellin, kinetin and indoleacetic acid, on seed germination and seedling growth of two strains of lettuce, cv. Attraktion and cv. Hohlblättriger Butter. (±)-Abscisin II inhibited seed germination and seedling growth in both strains. The inhibitory effect of abscisin II on seed germination as well as seedling growth was completely overcome by kinetin in both dark and light. Gibberellic acid, on the other hand, proved ineffective in exerting its influence on seed germination in presence of abscisin II but affected subsequent seedling growth as usual. Indoleacetic acid was found to be least potent in reversing abscisin II caused inhibition of seed germination and seedling growtb. It is concluded, that like its effect on certain other naturally occurring inhibitors of seed germination, kinetin can effectively undo the inhibitory effects of abscisin II in these growth processes.     

Characterisation of the High Temperature Inhibition of Germination of Lettuce (Lactuca sativa)

Lactuca sativa L., lettuce (Arctic King) fruit germination is inhibited in the light at temperatures above 27^°C(±1.5^°). This value, which is precise and repeatable is referred to as the upper temperature cut-off point, and is characteristic for a particular species. It was found, that this cut-off point is shifted upwards by applying kinetin solutions (0.1–1 mg/1) or downwards by applying (±)-abscisic acid solutions (over 1 mg/1), the amount of shift being proportional to the level of active compound. In the presence of both compounds a balance was achieved. Effects of stimulators and inhibitors can most explicitly be described by reference to the direction and magnitude of the shift in the upper temperature cut-off point, which results from their application.     

Effects of Temperature of Mother-plant Environment on Yield and Germination of Seeds of Lettuce (Lactuca sativa)

Experiments on the production of two separate crops of lettuceseeds, each in three different temperature environments, andsubsequent tests on the seed are described. Low production temperatures(20 °C day, 10 °C night) gave a low yield of large seeds,and high temperatures (30 °C, 20 °C) gave a higher yieldof much smaller seed; the highest yield came from medium temperatures(25 °C, 15 °C), which gave medium-sized seed. After-ripening,manifested as an increase in percentage germination at hightemperatures with increase in seed age, occurred in seed fromall three production environments of the first crop, thoughthere were differences in degree, and in that from the two higherproduction temperatures, but not the lowest, of the second crop.Measurements of the forces required to penetrate the layerssurrounding the embryo showed an inverse relationship with temperatureof the production environment for pericarps but not for endosperms,and a gradual reduction during storage for pericarps but notendosperms. Measurements of germination potential showed thatembryos from seeds produced in cool conditions were less ableto cope with high temperatures than those from warner conditions.These results are discussed in relation to the control of germinationin lettuce. Lettuce, Lactuca sativa (L.), seed production, germination, seed coverings, germination potential     

On the Structure of Lettuce (Lactuca sativa L.) Endosperm during Germination

Endosperm cells of lettuce (Lactuca sativa L.) are characterizedby thick cell walls and dense cytoplasm which contains numerousprotein bodies. Other organelles such as nucleus, mitochondria,plastids and dictyosomes are typical of plant cells. Light andelectron microscopy reveal that before radicle emergence micropylarcells of endosperm tissue undergo drastic protoplast alterations.These alterations seem to be the only structural modificationsbefore rupturing of the tissue since the walls of the endospermcells seem to degrade only after radicle emergence. The differentialbehaviour of the micropylar area of the endosperm before radicleemergence and the observation that the micropylar cells remainmetabolically active long after radicle emergence while therest of the tissue is almost completely disintegrated, suggeststhat the endosperm cells of the micropylar area may have a roleother than being a main reserve site like the rest of the endosperm. Lactuca sativa L., endosperm structure, seed germination, lettuce     

Interactions between Abscisic Acid, Gibberellins and Cytokinins in Grand Rapids Lettuce Seeds Germination

Experiments with Grand Rapids lettuce seeds (Lactuca sativa L.) maintained in darkness or irradiated with red light have shown that the inhibition of germination induced by low concentrations of ABA (2, 4, 6 μM) could be overcome by gibberellins (GA₃ or GA₄). The same results were obtained, although to a lesser extent, under the influence of two out of the four cytokinins tested (K and BAP) for seeds maintained in darkness. To suppress the block induced by higher concentrations of ABA (for example 8 μM), it was necessary to apply a cytokinin (K, BAP, Z or 2iP) and a gibberellin (GA₄ or GA₃) simultaneously, or a cytokinin following a red light treatment. Experiments conducted in darkness in which ABA (8 μM) was applied together with a cytokinin (BAP) and a gibberellin (GA₄) showed that the gibberellin and the cytokinin played similar roles towards each other and towards ABA.     

Influence of Different Cytokinins on the Germination of Lettuce (Lactuca sativa) and Celery (Apium graveolens) Seeds

The naturally occurring cytokinins, zeatin, zeatin riboside and dihydrozeatin did not promote the germination of celery (Apium graveolens L.) seeds and 6-Δ²-isopentenyladenine (2iPA) and its riboside were only moderately active. Of the synthetic cytokinins, kinetin, kinetin riboside, and the disubstituted urea, N-phenyl-N′-pyridyl urea (NC5392) were moderately active, and 6-benzyl-aminopurine (BA) and its derivatives BA riboside and 6-benzyl-amino-9(tetrahydropyran-2yl)purine (SD8339) were the most active cytokinins tested. 6-(o-hydroxybenzyl)aminopurine (hyd-BA) and its naturally occurring riboside inhibited germination under normally inductive conditions. All the cytokinins examined were more active in promoting germination of lettuce (Lactuca sativa L.) than celery seeds. BA, BA riboside and SD8339 were again the most active cytokinins. In contrast to the results with celery, zeatin and zeatin riboside were highly active. The other cytokinins also showed high activity with the exception of dihydrozeatin, hyd-BA and hyd-BA riboside which were less active. Cytokinin ribosides were less active than the corresponding free bases during the early period of the lettuce seed incubation but total germination after 90 h was similar.     

Release of Esterase Following Germination of Lettuce Seed (Lactuca sativa L.)

Light-insensitive lettuce seeds, Lactuca sativa L. cv. Great Lakes, release esterases for a period following radicle protrusion. Very little or no enzymes are released prior to 24 hours or after 48 hours of germination. As compared to intact seeds, half-seeds readily release esterases and the release is not affected by far red irradiation. Bulk of the released esterases are derived from the endosperm tissue and presumably exists in the intact seed as a component of the extraembryonic fluid.     

Characterization of the Germination Responses to Temperature of Lettuce (Lactuca sativa L.) Achenes

Germination tests were done on 23 cultivars of lettuce (Lactucasativa L.) comprising a variety of different morphological formsselected for cultivation at various seasons. Significant differences at the upper limit of temperature tolerancewere found between different cultivars: maximum temperaturesfor 50 per cent germination ranged from 23 to 32 °C, andsusceptibility to the induction of secondary dormancy by hightemperatures varied widely from one cultivar to another. Nocorrelations were established between germination responsesand heading type, achene colour or growing season The germination responses of all cultivars at temperatures fromjust above 0 to 20 °C were closely similar and results froma large number of experiments were used to produce a standardgermination reference curve foti culvars of L. sativa. Departfuresrom this curve were found to arise predominantly from variationsin seed quality or test conditions rather than from the genotypeof the cultivar. Detailed comparisons of germination at all possible combinationsof alternate and constant day/night temperatures are presentedfor two cultivars. One with relatively high tolerance to high-temperatureinducedinhibition, the other with low tolerance. The results are discussed in relation to the original distributionof the species as a wild plant and its history of cultivationby man. Lactuca sativa L., Lettuce, achene germination, temperature response     

莴苣胚囊细胞分离

用酶解和解剖方法分离了莴苣的卵细胞,助细胞,中央细胞和合子。莴苣子房先在酶液中酶解40～50min,然后在不含酶的分离液中用解剖针解剖子房。在解剖出的胚囊中,可看到卵细胞,两个助细胞和中央细胞的轮廓。将胚囊的合点端切破,轻轻挤压胚囊的珠孔端,四个细胞即可逸出。在最佳条件下,90min可从40个子房中分离出29个胚囊,进一步从中分离出11个卵细胞。分离出的胚囊细胞用显微操作仪收集备用。莴苣卵细胞的成功分离为进行离体受精探索创造了条件。     

Effect of Gibberellic Acid, Kinetin, and Ethylene plus Carbon Dioxide on the Thermodormancy of Lettuce Seed (Lactuca sativa L. cv. Mesa 659)

The effects of gibberellic acid and kinetin with ethylene plus carbon dioxide on the thermodormancy of lettuce seeds (Lactuca sativa L. cv. Mesa 659) at 35 C in the dark were studied. The combination of gibberellic acid plus kinetin with ethylene plus carbon dioxide was most effective in overcoming thermodormancy in these Great Lakes type seeds, alleviating any induced light requirement. Gibberellic acid action required at least a minimal level of ethylene plus carbon dioxide. Kinetin action was independent of ethylene plus carbon dioxide but interacted with the gases when the gases were added. A schematic representation of the interaction is presented.     

Effects of Germination-promoting Substances Given in Conjunction with Red Light on the Phytochrome-mediated Germination of Dormant Lettuce Seeds (Lactuca sativa L.)

Ethylene or thiourea can substitute for gibberellic acid but not for red light in breaking the secondary dormancy induced by extended dark storage of fully hydrated lettuce seeds (Lactuca sativa cv. Grand Rapids). After 10 days of storage, ethylene, thiourea, or gibberellic acid applied either separately or in any combination in conjunction with red light induced near maximal germination. When applied separately without red light, none of the substances promoted germination of seeds stored 10 days. Combinations of any two or all three of the substances in the absence of red light induced some germination but no combination was as effective as any single substance given with red light.     

Occurrence of Amoebae on Oak Leaf Lettuce (Lactuca sativa var. crispa) and Boston Lettuce (L. sativa var. capitata)1

The colonization, distribution, population density, and species diversity of amoebae on leaves of Oak Leaf lettuce, Lactuca sativa var. crispa, and Boston lettuce, L. sativa var. capitata, were investigated. The role of soil in the colonization of Oak Leaf lettuce was determined by comparing numbers of amoebae present on basal leaves (those that pass through soil) with numbers on wrapped leaves (those that do not pass through soil). Amoebae were present in ten samples of basal leaves and ranged from 154–1510/g of leaf tissue. Wrapped leaves failed to yield amoebae in seven of ten trials and contained <4 amoebae/g of tissue. Mean values for the population density of amoebae on Oak Leaf basal leaves and Boston lettuce leaves were 484 ± 133 and 453 ± 93, respectively. The distribution of amoebae on green and white parts of leaves from both kinds of lettuce was studied. The occurrence of amoebae on rinsed, unrinsed, visibly clean, and visibly dirty samples of Boston lettuce leaves was established.     

The Effect of Arsenate and Other Inhibitors on Early Events during the Germination of Lettuce Seeds (Lactuca sativa L.)

The effect of arsenate, arsenite, 2,4-dinitrophenol, and anaerobiosis on early events in seed germination was investigated using both intact and punched seeds of lettuce (Lactuca sativa L.). It was found that punching the seed removes penetration barriers to the entrance of inhibitors without an undue loss of germination or light responses. The kinetics of the action of germination inhibitors were established by 2-hour pulse experiments. Arsenate and 2, 4-dinitrophenol have very different kinetics. The inhibition of germination in punched seeds by arsenate given in conjunction with phosphate compared with the lack of inhibition of arsenate plus phosphate on the growing seedling, suggest a distinct metabolic change in the germinating embryo at some time between the onset of germination and subsequent seedling growth.     

Abscisic Acid Is an Endogenous Inhibitor in the Regulation of Mannanase Production by Isolated Lettuce (Lactuca sativa cv Grand Rapids) Endosperms

The production of mannanase, a cell-wall-degrading carbohydrase, can be manipulated in isolated lettuce (Lactuca sativa cv Grand Rapids) endosperms by changes in the volume of buffer in which they are incubated. The enzyme is produced when endosperms are incubated in a large volume, but not when incubated in a small volume, which is suggestive that an endogenous, diffusible inhibitor of mannanase production is being lost from the endosperms in a large volume (JD Bewley, P Halmer 1980/1981 Israel J Bot 29: 118-132). We have investigated the possibility that the phytohormone abscisic acid (ABA) is involved in this regulation of mannanase production in isolated lettuce endosperms. We find several correlations between the presence of the endogenous inhibitor and of ABA, i.e. (a) a `leachate' prepared from isolated lettuce endosperms induces synthesis of ABA-specific proteins in barley aleurone layers, indicating that incubation of endosperms in a large volume results in the diffusion of ABA therefrom into the surrounding medium; (b) fractionation of the components of a leachate by either polyvinylpyrrolidone-chromatography of C₁₈ reversed-phase high performance liquid chromatography fails to separate the endogenous inhibitor from authentic ABA; and (c) changes in the incubation volume of endosperms result in changes in the amount of extractable ABA in the endosperms, as detected by ELISA. These results are consistent with a role for endogenous ABA in the regulation of mannanase production in isolated lettuce endosperms.     

Influence of After-ripening on Phytochrome Control of Seed Germination in Two Varieties of Lettuce (Lactuca sativa L.)

The reversible photoreaction on seed germination of two varieties of lettuce differs remarkably not only with the variety but also with the germination temperature and the physiological conditions of seeds caused by after-ripening.     

Interactions between Mild NaCl Stress and Red Light during Lettuce (Lactuca sativa L. cv Grand Rapids) Seed Germination

The sensitivity of lettuce (Lactuca sativa L. cv Grand Rapids) seeds to red light was reduced by NaCl concentrations which had no effect upon the germination of continuously illuminated seeds. The germination capacity of the seeds was fully restored by increased red light exposures. Indirect evidence indicates that NaCl does not affect the photoconversion of red-absorbing form of phytochrome to the far-red absorbing form of phytochrome. Instead, the increased red light requirements are attributable to increases in the threshold levels of the far-red absorbing form of phytochrome necessary to induce germination and to changes in the slopes of the fluence-response curves. Results also show that the sensitivity of the seeds to NaCl decreased as the time between red light irradiation and the imposition of NaCl stress increased.     

Germination and growth of lettuce (Lactuca sativa) at low atmospheric pressure

The response of lettuce ( Lactuca sativa L. cv. Waldmann's Green) to low atmospheric pressure was examined during the initial 5 days of germination and emergence, and also during subsequent growth to vegetative maturity at 30 days. Growth took place inside a 66-l-volume low pressure chamber maintained at 70 kPa, and plant response was compared to that of plants in a second, matching chamber that was at ambient pressure (approximately 101 kPa) as a control. In other experiments, to determine short-term effects of low pressure transients, plants were grown at ambient pressure until maturity and then subjected to alternating periods of 24 h of low and ambient atmospheric pressures. In all treatments the partial pressure of O₂ was maintained at 21 kPa (approximately the partial pressure in air at normal pressure), and the partial pressure of CO₂ was in the range 66.5–73.5 Pa (about twice that in normal air) in both chambers, with the addition of CO₂ during the light phase. With continuous exposure to low pressure, shoot and root growth was at least as rapid as at ambient pressure, with an overall trend towards slightly greater performance at the lower pressure. Dark respiration rates were greater at low pressure. Transient periods at low pressure decreased transpiration and increased dark respiration but only during the period of exposure to low pressure. We conclude that long-term or short-term exposure to subambient pressure (70 kPa) was without detectable detriment to vegetative growth and development.     

适用于生菜叶片蛋白质双向电泳方法的建立及初步应用

以生菜(Lactuca sativa)叶片为研究材料,参考拟南芥和水稻的双向电泳方法,分析了等点聚焦时间和染色方法等影响双向电泳的关键因素,建立适合生菜叶片总蛋白质分离的双向电泳方法:采用三氯乙酸-丙酮沉淀法进行总蛋白质提取,用24cm固相pH梯度等电聚焦8000V×8h结合12%的SDS-PAGE进行双向电泳分离,银染法和考马斯亮蓝染色法染色都获得了较好的结果.应用Image-Master-Elite软件对考马斯亮蓝染色法染色的图谱进行分析表明:每张凝胶上都检测到超过1000个的蛋白质,不同凝胶之间蛋白质的匹配律达到98%,具有较高的分辨率和重复性.初步分析了生菜叶片蛋白质的等电点、分子量的分布情况,发现生菜叶片蛋白质的等电点以5.0～5.5之间最多而分子量主要集中在20～60kD之间.     

Metabolism of (±) 2-[14C]-abscisic acid in Lactuca sativa achenes

Achenes ofLactuca sativa L. cv. Grand Rapids were treated with (±) 2-[¹⁴C]-abscisic acid (ABA) at 10⁵ - or 2-10⁶ M for 6, 12, 24, 48 or 96 h in darkness at 24°C. They were then extracted in 80% ethanol. Two acidic diethyl ether phases which contained the free acids and the acids released after mild alkaline hydrolysis respectively, were analyzed as well as the radioactivity which remained in the final aqueous phase. For treatment durations between 6 and 96 h, the major part of the radioactivity was found in the free phase, in the form of ABA. For treatment durations up to 48 h, no radioactivity was detected at the Rf of phaseic acid or dihydrophaseic acid (free and hydrolysed phases). After 96 h culture on 10⁵ M ABA, dihydrophaseic acid was present, but only in very small quantities. Two ABA metabolites were detected. One was characterized as β-d -glucopyranosyl abscisate since its Rf was the same as that of an authentic sample in three different solvent systems and also since it released ABA on mild alkaline hydrolysis. It increased steadily with time and represented the main metabolite. The other metabolite found in the aqueous phase after mild alkaline hydrolysis and extraction with ether at pH 3 was a very polar compound, resistant to alkaline hydrolysis in the presence of concentrated ammonia and to methylation. It was, however, metabolized by apple embryo, yielding essentially dihydrophaseic acid and an ester which released dihydrophaseic acid on mild alkaline hydrolysis. These results indicate that under the conditions tried, the metabolism of [¹⁴C]-ABA by lettuce achenes leads almost exclusively to the formation of conjugates, oxidative metabolism of ABA being almost non-existent. Separate analysis of the integuments and of the endosperm plus embryo after culture of whole achenes for 48 h in the presence of 10⁵ M [¹⁴C]-ABA showed that ABA metabolism occurred only in the endospermembryo tissue.