Seed germination at different temperatures and seedling emergence at different depths of Rhamnus spp

Rhamnus alaternus and R. ludovici-salvatoris, two Mediterranean shrubs with different geographic distributions, have shown important differences in seedling recruitment capacity. The objectives of this work were to determine the ability of these species to germinate seeds under different temperature ranges, as well as the capacity of seedlings to emerge from different burial depths, in order to better understand their regeneration processes. Two different experiments were performed. In the first one, seed germination was studied in Petri dishes and in the dark at different temperature regimes: a) 5–15°C, b) 10–20°C and c) 15–25°C (12h/12h). In the second experiment, seedling emergence capacity from different burial depths (0.5, 2 and 5 cm) was tested. R. ludovici-salvatoris showed a significantly higher final germination rates, a lower dormancy period, and average time response at 10–20°C than at other temperature ranges, although differences were much greater when seeds were subjected to the 5–15°C temperature regime. By contrast, R. alaternus did not show significant differences between treatments (5–15°C and 10–20°C) in germination behavior. Seedling emergence of both species was lower and slower when seeds were buried at 5 cm. However, R. ludovici-salvatoris always showed a lower seedling emergence capacity than R. alaternus at any burial depth. The low ability of R. ludovici-salvatoris to germinate seeds and emerge between 5–15°C, even from shallow depths, is discussed in relation to its low regeneration capacity and declining geographic distribution.     

Respiration and germination rates of tomato seeds at suboptimal temperatures and reduced water potentials

Suboptimal temperature (T) affects germination rates (reciprocalof time to radicle emergence) on a thermal time basis; thatis, the T in excess of a base or minimum temperature multipliedby the time to a given per cent germination [t_g) is a constant.Respiration rates are also sensitive to T, and proportionalrelationships are often found between respiration rates andgermination rates. Reduced water potential () delays seed germinationon a hydrotime basis (i.e. the in excess of a base water potentialmultiplied by t_g is a constant). It was tested whether respirationrates prior to radicle emergence vary in proportion to T and as expected from the thermal and hydrotime models. Respirationrates (C0₂ evolution) of cold-tolerant, rapidly germinating(PI 341988) and cold-sensitive, more slowly germinating (T5)tomato [Lycopersicon esculentum Mill.) seeds were evaluatedover a range of T and conditions. For both genotypes, respirationrates until the beginning of radicle emergence were relatedto T on a thermal time basis and increased approximately linearlywith above -2.0 MPa, consistent with the hydrotime model. Respirationrates were uniquely related to germination rates, regardlessof whether germination timing was affected by T, , or genotype.However, germination timing was unaffected when respirationrates were manipulated by varying 0₂ partial pressure. Thus,while both germination and respiration rates vary with T and consistent with thermal and hydrotime models of biologicaltime, respiration rates per se were not the limiting factorin germination timing of tomato seeds. Key words: Lycopersicon esculentum Mill., tomato, germination, respiration, temperature, water potential     

The viability and infectivity ofToxocara canis infective larvae after a prolonged period of storage at different temperatures

Summary Larvae ofToxocara canis were developed and stored at different temperatures for different periods of time at 70 to 80% relative humidity. After 18 months the infectivity of these larvae was tested byin vitro hatching and by the use of white mice (in vivo infection). Although the larvae were stored for very long periods, there was no indication that they were weaker than the control sample. After a further 7 months, when part of the original sample was exposed to higher temperatures (35 to 41°C), a high proportion of them (70%) hatched spontaneously and died, but the larvae of the remaining 30% were still viable and active and forming a source of danger to the community.

---

Resumen Larvas deToxocora canis se desarrollaron y guardaron a differentes temperaturas durante distintos periodos de tiempo a 70–80% de humedad relativa. Alos 18 meses se probó la infectividad de dichas larvasin vitro y utilizando ratones blancos (infecciónin vivo). Aunque las larvas se hubieran almacenado durante largos periodos no se observó que fueran más débiles que la muestra control. Pasados otros 7 meses, cuando parte de la muestra original se expuso a temperaturas más elevadas (35 a 41°C) gran parte de las larvas (70%) eclosionaron espontaneamente y murieron, sin embargo las larvas restantes (30%) siguieron viables y activas representando una fuente de peligro para la comunidad.

---

Résumé On a développé des larves deToxocara canis et on les a conservées à différentes températures pendant différents temps à une humidité relative de 70 à 80%. Après 18 mois, le pouvoir infectieux de ces larves a été testé par incubationin vitro et à l'aide de la souris blanche (infectionin vivo). Bien que les larves aient été conservées pour de longues périodes, il n'y a eu aucune indication de ce qu'elles étaient plus faibles que celles de l'échantillon de contrôle. Après 7 mois supplémentaires, alor qu'une partie de l'échantillon de départ était exposé à des températures plus élevées (37 à 41°C), une proportion élevée de ces larves (70%) a éclos spontanément et en est morte, mais les 30% restant étaient toujours viables et actifs, formant ainsi une source de danger pour la communauté.

     

汉麻籽不同萌发时期的生长活性物质非靶向代谢组学研究

为了使汉麻籽在食品工业中发挥其最大价值,找寻汉麻籽在萌发过程中的最佳处理时期十分重要。本研究利用非靶向代谢组学研究方法和液相色谱-质谱(LC-MC)联用技术分析汉麻籽在发芽12及24 h的代谢物数量,结合多元统计分析方法与单变量统计分析,筛选汉麻籽三个时期组间的显著性差异代谢物(未发芽与发芽12 h的汉麻籽,未发芽与发芽24 h的汉麻籽,发芽12 h与发芽24 h的汉麻籽)。结果表明在正、负离子监测模式下,三个时期共发现显著性差异代谢物26个,第一组中16种表现为上调,8种表现为下调;第二组中13种表现为上调,9种表现为下调;第三组中13种表现为上调,8种表现为下调。其中,在汉麻籽发芽12 h时二羟基丙酮(DHA),4-羟基丁酸内酯,4-氨基丁酸,嘧啶等表现为上调;D-脯氨酸,烟酸酯,L-脯氨酸等表现为下调,且其变化倍数远远大于其他成分,影响作用较明显。本研究为萌发汉麻籽在食品开发中的应用提供参考。     

Effects of long-term storage on phytochrome-mediated germination in lettuce seeds

The effects of long-term seed storage on the physiological properties of phytochrome-mediated germination including water uptake, the temperature and light flunnce dependencies of germination and dark germination were studied. The fluenceresponse relationships of the brief irradiation with monochromatic red (660 nm, 7.5 W m⁻²) and far-red (750 nm, 6.6 W m⁻²) light at various times after sowing were also studied. The samples used consisted of three lots of seeds ofLactuca sativa L. cv. MSU-16, which had been harvested in 1976, 1979 and 1985 and stored dry for 9, 6 and 0 years, respectively, in darkness at 23±2 C until the experiments were carried out in July–August, 1985. Seeds with the longer storage periods showed the higher ability to germinate in both continuous darkness and continuous white fluorescent light at 20–30 C. In the seeds stored for 6 or 9 years, red light irradiation for 20 sec given at 15 min or more after sowing at 25 C induced as high a percent germination (85–95%) as those under continuous white fluorescent light. In the freshly harvested seeds, however, germination under continuous white fluorescent light (46%) was considerably lower than the germination induced by the red pulse (97%). Germination of the seeds decreased when the intervals between sowing and a far-red irradiation for 20 sec increased up to 100 min (or 30 min in the freshly harvested seeds). The far-red pulse given later than 100 min (or 6 hr in the freshly harvested seeds) after sowing resulted in an increased germination up to the dark-germination levels with increasing intervals between sowing and the pulse irradiation. Before or at 3 min after sowing, the seeds stored for 6 or 9 years were responsive to the far-red pulse although they were not or hardly responsive to the red pulse, while the freshly harvested seeds were responsive to both the far-red and the red pulses. These data indicate that normal functions of phytochrome completely survived in the dry seeds during storage at 25 C for as long as 6 or 9 years and that these functions are restored into full operation by means of imbibition. The differences in the dependence of germination on the time and fluence of a single pulse of red or far-red light seems to be related to the smaller water content throughout the imbibition in the seeds with the longer storage periods. The greater ability to germinate in the dark indicates the greater amounts of P_FR or the greater responsivity to P_FR, in the seeds with the longer storage periods.     

不同贮藏温度下大别山五针松花粉活力的变化

大别山五针松( Pinus dabeshanensis C. Y. Cheng et Y. W. Law)为松科( Pinaceae)松属( Pinus Linn.)植物,自然种群数量极少,目前仅发现在安徽省岳西县大王沟海拔900~1300 m阴坡和半阴坡有相对集中的分布,种群规模计200余株,且多为成年个体,林下幼苗极少,自然更新困难,为中国特有珍稀树种之一[1]。大别山五针松常在每年3月份至4月份形成花芽,5月中下旬花粉成熟并散发,花粉具气囊,此时雌球花张开接受传粉,球果翌年9月成熟[2]。据作者近年的调查,大别山五针松种子败育率较高,且种子质量较低,这也是该种类濒危的重要原因之一。     

Proteolysis of the main storage protein of buckwheat seeds at the early stage of germination

The changes in the main storage protein of seeds of buckwheat ( Fagopyrum esculentum Moench cv. Shatilovskaya 5), 13S globulin, were studied during seed germination. During the first three days of germination the 13S globulin is subjected to a limited proteolysis, which consists in the splitting of some of its subunits into large polypeptide fragments. Insignificant changes in the sedimentation coefficient of the 13S globulin during the first days of germination as well as immunochemical data, indicate that the limited proteolysis of the 13S globulin does not cause any major changes in its structure.
Dormant buckwheat seeds contain a proteolytic enzyme (a metalloproteinase), which can cause limited proteolysis of 13S globulin. The proteinase hydrolyzed some subunits of the 13S globulin to high molecular weight fragments. In the presence of sodium dodecylsulphate the electrophoretic pattern of 13S globulin, isolated from 3-day-old buckwheat seedlings, was almost identical to that of 13S globulin from dormant seeds hydrolyzed with metalloproteinase. It is suggested that the proteolysis of 13S globulin observed in vitro may also take place in vivo in the course of seed germination.     

Prediction of the shelf-life of pasteurized milk at different storage temperatures

Initial psychrotroph counts determined by a Most Probable Number technique were correlated with shelf-lives of pasteurized milk determined at a number of storage temperatures. The initial psychrotroph count was also correlated with a bacterial count carried out on milk agar containing crystal violet penicillin and nisin after previous incubation of the milk at 15°C for 25 h.
Pre-incubation counts carried out at a variety of temperatures and on a variety of media were examined for their relation to shelf-life. Shelf-lives at four pre-set temperatures (2, 6, 10 and 14°C) could best be predicted by pre-incubation of pasteurized milk at 15°C before inoculation on milk agar.
An equation which allows prediction of shelf-life of pasteurized milk at any storage temperature is described.     

The effect of storage at three different temperatures on the activity of lipase solution

A useful practical investigation carried out by fifthformers at Sawston Village College     

Effect of ABA, fusicoccin and thiourea on germination and K+ and glucose uptake in chick-pea seeds at different temperatures

Germination and growth of seeds of Cicer arietinum L. are retarded or inhibited either by temperatures above 30°C or by the presence of abscisic acid (ABA). Both treatments affect the uptake of K⁺ and glucose. ABA may affect the properties of the cell membrane or the kinetics of the enzymes bound to it. Fusicoccin (FC) and thiourea (TU) are to a greater or lesser extent able to counteract the inhibition produced by ABA and high temperatures. Both FC and TU increase the uptake of K⁺ by the embryonic axis. FC seems to act directly on H⁺ extrusion, but the mode of action of TU appears to be different. The results seem to indicate the importance of the activation of the ion transport process for the growth of the embryonic axis during early germination, and suggest a control of the growth of the embryo by part of the cotyledons. Physiological inhibitors such as ABA might be implied in the control mechanism.     

Penetration ofCampylobacter jejuni at different storage temperatures in membranes and contents of cracked eggs

Summary Campylobacter jejuni 79-193 penetrates the inner, outer membrane and ultimately the contents of cracked eggs more at 4°C (after equilibration at 42°C for 10 min) than at 42°C or 25°C. The number of eggs showing penetration byC. jejuni decreased with time.

---

Pénétration de Campylobacter jejuni à différentes températures dans les membranes et contenus d'oeux félés

Résumé Campylobacter jejuni pénètre les membranes externe, interne et finalement le contenu d'oeux félés davantage à 4°C (après équilibration à 42°C pendant 10 min.) qu'à 25 ou 42°C. Le nombre d'oeux montrant la pénétration parC. jejuni décroît avec le temps.

     

Lipases in the storage tissues of peanut and other oil seeds during germination

The castor-bean endosperm-the best-studied material of reserve lipid hydrolysis in seed germination-was previously shown to have an acid lipase and an alkaline lipase having reciprocal patterns of development during germination. We studied oil seeds from 7 species, namely castor bean (Ricinus communis L.), peanut (Arachis hypogaea L.), sunflower (Helianthus annus L.), cucumber (Cucumis sativus L.), cotton (Gossypisum hirsutum L.), corn (Zea mays. L.) and tomato (Lycopersicon esculentum Mill.). The storage tissues of all these oil seeds except castor bean contained only alkaline lipase activity which increased drastically during germination. The pattern of acid and alkaline lipases in castor bean does not seem to be common in other oil seeds. The alkaline lipase of peanut cotyledons was chosen for further study. On sucrose gradient centrifugation of cotyledon homogenate from 3-d-old seedlings, about 60% of the activity of the enzyme was found to be associated with the glyoxysomes, 15% with the mitochondria, and 25% with a membrane fraction at a density of 1.12 g cm^-3. The glyoxysomal lipase was associated with the organelle membrane, and hydrolyzed only monoglyceride whereas the mitochondrial and membrane-fraction enzymes degraded mono-, di- and triglycerides equally well. Thus, although the lipase in the glyoxysomes had the highest activity, it had to cooperate with lipases in other cellular compartments for the complete hydrolysis of reserve triglycerides.     

Survival of Vibrio cholerae 01 in common foodstuffs during storage at different temperatures

Survival of Vibrio cholerae El Tor serotype Inaba was examined in pasteurized milk, freshwater fish, raw beef and raw chicken at a variety of temperatures. Both food type and incubation temperature affected survival. At the lowest temperatures, V. cholerae remained viable in meats for up to 90 d at—5°C and 300 d at —25°C. In milk, however, it was not detectable after 34 d at —5°C and 150 d at —25°C. At 7°C it survived 32 d, on average, in milk and only 18–20 d in the other foods. At room temperatures survival periods were shorter, never exceeding 10 d, and it was not detected after 2 d incubation at 35°C in chicken and fish.     

Viability and virulence of blastospores of Metarhizium anisopliae (Metch.) Sorokin after storage in various liquids at different temperatures

Blastospores of three strains of Metarhizium anisopliae were stored in 18 liquids at 4°C, 20°C and 35°C for 18 weeks, 12 weeks or 9 days respectively. Viability was quantified by determination of their germination. In bioassays the virulence of stored blastospores was studied using adults and third instars of Locusta migratoria migratorioides (R. & F.) and compared to those of freshly produced blastospores and conidia. Generally, there was great variability in the viability of blastospores, depending on the fungal strain and the liquids used. Blastospores survived best at 4°C in 10% hydroxyethyl starch; for example, germination of M. anisopliae strain 97 still amounted to more than 80% after storage for 18 weeks. Other suitable liquids were deionized water, 25% Ringer's solution and 1% sodium alginate. The viability of blastospores stored at 20°C was considerably shorter than at 4°C. During storage for 12 weeks at 20°C the best protective liquids for M. anisopliae strain 97 were 25% Ringer's solution (43% germination), deionized water (23%) and 10% hydroxyethyl starch (23%). At 35°C, 45% of M. anisopliae strain 97 blastospores still germinated after storage for 7 days in 25% glycerol. The bioassays revealed that the virulence of blastospores after storage was comparable to that of fresh ones and even better than that of fresh conidia. In general, the LT₅₀ was about 4–6 days at an alternating day/night temperature of 28/20°C.     

Endoreduplication in cucumber (Cucumis sativus) seeds during development, after processing and storage, and during germination

Flow cytometry was used to study endoreduplication in developing, stored and germinating seeds of cucumber ( Cucumis sativus ). Fruits growing in a commercial seed production field were collected every 7 days, starting 14 days after pollination (DAP) up to 63 DAP (commercial harvest time). Seeds were isolated and the proportion of nuclei with different DNA contents in the whole seeds and in the embryos was analysed. Germination capacity of fresh and dried seeds at 25°C was established. In addition, the same analyses were performed on the seeds after processing (fermentation, drying and cleaning), following 1 and 2 years of storage, and after imbibition for 3, 6 and 12 h. In the young developing seeds, endoreduplication up to 128C occurred but this decreased to 8C by maturity. The proportion of endosperm nuclei was the highest at 21 DAP (30%) and then decreased to below 14% at harvest and 8% after processing. In the mature processed seeds, the majority of embryo nuclei (about 80%) contained 2C DNA; however, about 2% of endoreduplicated (8C) nuclei were still present. Seeds did not show any germination capacity up to 21 DAP; then it gradually increased to reach 100% as early as 49 DAP, 2 weeks before commercial harvest time. The relationship between seed maturity, germination and cell cycle status is discussed. The mean C-value of the seed cells as well as the (4C + 8C + 16C)/2C ratio are recommended as markers of cucumber seed maturity and the advancement of germination/priming (the stage of germination sensu stricto ).     

Effects of storage on germination ofDioscorea composita (Dioscoreaceae) seeds

Effects of storage were tested on germination ofDioscorea composita (Dioscoreaceae) seeds. Freshly collected seeds and seeds stored at 25°C in paper bags from 1 to 11 mo or for 4 and 5 yr were used in most of the experiments. Seeds were tested for germination at 20, 25, 30, 35, 25–20, and 25–35°C in white light and in darkness. Initiation of germination was delayed in freshly harvested seeds, and dormancy was reduced in seeds stored for about 9 mo. Viability of the seeds decreased after 4 and 5 yr of storage.