Dormancy Breaking and Storage Behavior of Garcinia cowa Roxb. (Guttiferae) Seeds: Implications for Ecological Function and Germplasm Conservation

The dormancy breaking and storage behavior of Garcinia cowa Roxb. seeds were investigated.The seeds of G. cowa had 8-11 months dormancy in their natural habitat. Seeds were matured and dispersed at the end of the rainy season (mid-late August to late September) and were scatter-hoarded by rodents as food for winter after the seeds had fallen to the ground. Seedlings often emerged in the forest during the rainy season (May to August) the following year. Intact seeds of G. cowa failed to germinate after being sown at 30 ℃ for 120 d and the mean germination time (MGT) of seeds cultured in a shade (50% sunlight)nursery was 252 d. The most effective method of breaking dormancy was to remove the seed coat totally,which reduced the MGT to 13 d at 30 ℃. Germination was also promoted by partial removal of the seed coat (excising the hilum and exposing the radicle) and chemical scarification (immersion in 1% H2O2 for 1 d).Unscarified seeds take up water rapidly in the first 96 h, but water was absorbed by the outside seed coat,without penetrating through it. The moisture content (MC) of G. cowa seeds was high (50% in fresh weight)at shedding. The seeds could tolerate desiccation to some extent, until the MC reached approximately 40%;below that, the viability decreases rapidly and all seeds died at approximately 17% of MC. Seed viability decreased rapidly when seeds were chilled at 4 ℃; germination was 2% after storage for 1 week. Even stored at 10 ℃, seeds began to be damaged after 4 weeks. Seed storage for 1 yr revealed that in both dry (relative humidity (35 ± 5)%) and moist (wet sand) storage conditions, seed viability declined, but germination percentages for seeds stored under moist conditions are better than for seed stored under dry conditions.Because of their low tolerance to desiccation, marked chilling sensitivity and relatively short lifespan, G.cowa seeds should be classified into the tropical recalcitrant category. The ecological implications of dormant recalcitrant seeds and cues on storing recalcitrant seeds were discussed.     

Artificial seeds of alfalfa (Medicago sativa L.). Induction of desiccation tolerance in somatic embryos

Summary The use of somatic embryos from cell culture systems in the clonal propagation of plants would be greatly facilitated if the somatic embryos could be dried and stored in a dormant state similar to true seeds. A cell culture system was developed for alfalfa (Medicago sativa L.) line RL34 which gave high yields of somatic embryos in an approximately synchronized pattern. These somatic embryos were treated with abscisic acid (ABA) at the cotyledonary stage of development to induce desiccation tolerance. With no visual preselection, approximately 60% of the dried embryos converted into plants upon reimbibition. When high quality embryos were selected prior to drying, 90 to 100% conversion rates were observed. The timing of the application of ABA in terms of embryo development was critical with an optimum being at cotyledonary stage spanning approximately 4 days; thus, synchronized embryo development is required for optimal expression in bulk samples. The vigor of the seedlings from dried somatic embryos was greater than those from embryos which had not been dried, but remained substantially lower than those from true seeds.     

Effect of day length on germination of seeds collected in Alaska

Day length control can effectively limit seed germination to favorable seasons, but this phenomenon has been studied in relatively few wild plants. I tested species from interior Alaska for day length control of germination under controlled conditions, and I also monitored germination phenology in natural habitats. Unstratified and cold-stratified seeds were germinated on short (13 h) and long (22 h) day length and in the dark at constant and alternating temperatures. On long day length, unstratified Ledum decumbens and Saxifraga tricuspidata seeds germinated from 5 C to 20 C, but on short day length few or no seeds germinated at 5 C and 10 C and germination was reduced at higher temperatures. Unstratified seeds of Diapensia lapponica and Chamaedaphne calyculata germinated only at 15 C and 20 C on long day length, and short day length completely inhibited germination. Cold stratification widened the temperature range for germination on both long and short day lengths, but germination was still lower on short than long day length. Germination phenology in natural habitats was consistent with germination in controlled conditions. In these species, short day length and low temperatures interact to inhibit germination in the fall. After overwintering, seeds germinate in the spring at low temperatures and on long day lengths. The inhibitory effect of short day length is not important in the spring because day length is already long at snowmelt.     

箭叶淫羊藿种子的休眠类型与萌发研究

箭叶淫羊藿具有重要的药用价值和较大的市场需求,但其种子休眠和萌发特性目前仍不够清楚,严重影响了其产业化育苗与栽培。为探明其种子的休眠类型及释放休眠和促进休眠的最适方法,该文以箭叶淫羊藿的成熟种子为材料,研究种子对水分的吸收、种子的脱水耐性,以及温度、层积和植物激素对种子休眠与萌发的影响。结果表明：(1)箭叶淫羊藿种子不具有物理休眠,而具耐脱水性。(2)在4～25℃和黑暗条件下,种子萌发率为零,具有休眠特性。(3)胚与种子的比值非常小,4℃和10℃及变温层积能显著地促进胚的生长发育以及增加种子的萌发速率和萌发率。(4)赤霉素和氟啶酮显著增加种子的萌发速率和萌发率。综上认为,箭叶淫羊藿种子的休眠类型为形态生理休眠(MPD),释放休眠和促进萌发的最适方法是先将种子在10℃中层积30 d,再置于4℃环境中让其萌发。该研究结果为箭叶淫羊藿的产业化育苗提供了参考。     

假槟榔种子催芽技术和脱水耐性的研究

为提高假槟榔的人工种植技术,对其种子做了不同的化学催芽处理,以寻求种子的有效催芽方法,并对种子脱水耐性进行了探讨。结果表明:20%过氧化氢和98%浓硫酸浸泡5min,0.3%亚硝酸钠和0.2%硝酸钾溶液浸种24h后,发芽率显著升高,速度显著加快,尤以浓硫酸和硝酸钾处理效果为好;200～1000mg/L赤霉素和20～100mg/L激动素溶液浸泡24h也显著促进种子萌发,但催芽效果与溶液浓度有关。成熟种子轻度脱水,发芽率有所上升,但含水量下降至17%以下,发芽率急剧下降,当含水量下降10%以下,发芽力完全丧失。由此可见,种子很可能是中间型种子。     

沙芥种子发育过程中的脱水耐性

以发育过程中经脱水和未脱水处理的沙芥种子为试验材料,测定了其含水量、萌发率和抗氧化酶系统,探讨了沙芥种子脱水耐性与抗氧化系统之间的关系。结果表明：在20～60DAF,沙芥种子含水量逐渐下降,干重逐渐增加;60DAF种子具有萌发能力,萌发率为24％;且脱水可促进沙芥种子的萌发,人工脱水至含水量为12％和5％时,萌发率分别为56％和44％,自然脱水至含水量为12％和5％时,萌发率分别为52％和60％;发育过程中沙芥种子SOD活性逐渐降低,而CAT、POD、LOX活性以及MDA含量均呈上升趋势;在脱水过程中,随着种子含水量的下降SOD活性逐渐降低,CAT和LOX活性逐渐升高,而POD活性呈先降低后升高的变化趋势。脱水后,种子中MDA含量均高于CK。60DAF的沙芥种子已获得脱水耐性。     

Intraspecific differences in the response to drying of Quercus ithaburensis acorns

In this study, we investigated the germination behavior, and the differences in desiccation sensitivity and water uptake (WU), of the acorns of the two subspecies of Quercus ithaburensis, inhabiting different ecological niches. Sensitivity of acorns to drying was assessed by subjecting them to several desiccation treatments. The results showed that acorn water loss due to desiccation follows an almost linear pattern for the subspecies macrolepis, while in the case of subspecies ithaburensis, it follows a negative exponential model. Accordingly, the germination behavior was found highly dependent on water loss for both subspecies. The low acorn water content (15%) achieved after 15 days of drying was crucial for the ssp. macrolepis, while it had no effect on seed viability in ssp ithaburensis, that needed a subtle further water loss to reduce its germinability (i.e. <14.5% mc). Our results suggest that intraspecific differences in seed responses to drying may be due to different adaptation to survive drought after dispersal and contribute to explain the ability of ssp. ithaburensis to grow in the hottest and driest Mediterranean bioclimates. Despite being recalcitrant, the ssp. ithaburensis can be considered as one of the most tolerant oaks to desiccation worldwide.     

油菜种子发育和萌发过程中脱水耐性相关基因的cDNA-AFLP分析

选取甘蓝型油菜种子发育和萌发过程中与脱水耐性相关的不同时间点,采用cDNA-AFLP技术通过128对引物组合分析,共获得394条特异转录衍生片段(TDFs)。经BLAST序列比对,发现其中189条与GenBank数据库中已知功能基因同源,按其功能可分为转录因子、基础代谢、抗逆胁迫、信号转导等相关蛋白。RT-PCR技术对差异显著性片段验证结果显示,泛素蛋白、油体蛋白、热激蛋白和一些转录因子片段均受到种子脱水诱导表达。研究结果表明,发生在种子发育和萌发过程中的脱水耐性反应非常复杂,涉及植物生命活动的诸多领域。     

Age-related changes in antioxidant enzymes and prooxidant generation in tissues of the rat with special reference to parameters in two insect species

The objective of this study was to explore the relationship between partially reduced oxygen species and the aging process. Effect of age on antioxidant defenses and prooxidant generation was evaluated by comparing the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase, and rates of mitochondrial O-₂ and H₂O₂ generation in the liver, heart, and brain of 3-month and 18-month-old Sprague-Dawley rats. In addition, comparisons of antioxidant defenses and mitochondrial prooxidant generation were made between short-lived insects and the rat tissues. Results indicated that antioxidant enzymes exhibit a mixed pattern of age-related alterations. In each organ of the rat examines, activities of some enzymes were up with age and of others down with age. The overall magnitude of decline in antioxidant defenses observed here and elsewhere in the literature was deemed to be unlikely to be functionally significant. In contrast, the rate of mitochondrial O-₂ and H₂O₂ generation increased in various tissues of the rat. Antioxidant defenses in insects were comparable to tissues in the rat but rates of O-₂ and H₂O₂ generations were notably higher. Results are interpreted to suggest that rates of prooxidant generation may be more crucial than antioxidant levels as possible longevity determinants.     

小芸木胚发育过程中脱水耐性的变化

对热带植物小芸木的种子和胚在整个发育阶段的形态学特征、含水量、萌发率和电导率进行了研究。结果显示:(1)小芸木种子在发育过程中形态学特征、电导率、萌发率和胚的脱水耐性有明显的变化;(2)在55~80 d种子鲜重和干重逐渐增加,随后又稍微降低,整个发育过程未经历明显的成熟脱水阶段;种子萌发率从55~85 d逐渐达到最大,随后又稍微降低;(3)胚的脱水耐性从55~90 d逐渐增加,于85~90 d达到最大,在95~103 d时又有所下降。表明小芸木胚的最大脱水耐性的获得时间与种子干物质积累达到最大的时间一致。     

Homoiohydrous (recalcitrant) seeds: Developmental status,desiccation sensitivity and the state of water in axes of Landolphia kirkii Dyer

The desiccation sensitivity in relation to the stage of development was investigated in embryonic axes from the homoiohydrous (recalcitrant) seeds of Landolphia kirkii. Electrolyte leakage, used to assess membrane damage after flash (very rapid) drying, indicated that axes from immature (non-germinable) seeds were the most desiccation-tolerant, followed by those from mature seeds, while axes from seeds germinated for increasing times were progressively more desiccation-sensitive. Differential scanning calorimetry was used to study the relationship between desiccation sensitivity and the properties of water in the tissues. Axes from immature seeds had a lower content of non-freezable water than that of any other developmental stage and a higher enthalpy of melting of freezable water. For mature and immature axes electrolyte leakage increased at the point of loss of freezable water. At other developmental stages the water content at which electrolyte leakage increased markedly correlated with the other properties of the water, such as the change in the shape of the melting endotherm and the onset temperature. Ultrastructural studies of axes at the various developmental stages showed changes in the degree and pattern of vacuolation, the presence and quantities of lipid and starch, and the degree of endomembrane development. The results are discussed in relation to current hypotheses on the basis of desiccation tolerance.Abbreviation DSC differential scanning calorimetry     

Dormancy Release,Germination, and Electrolyte Leakage from Apple Embryos during Stratification in the Presence of Sucrose

The dynamics of dormancy release during the stratification of apple (Malus domestica Borkh.) seeds was quantitatively described by three characteristics of seeds germination: the percentage of seeds that germinated by the tenth day, mean germination time, and the sum of seeds germinated in each of ten days (Timson's parameter), which allowed the assessment of the viability, the rate of dormancy release, and seed heterogeneity. We showed that apple seeds were characterized by a combined (physical and physiological) type of dormancy, with the seed coat and the embryo envelope being involved in the maintenance of physical dormancy. The addition of sucrose to the stratification medium accelerated the release of seed dormancy and improved all characteristics that determine seed germinability. Electrolyte leakage from embryos hardly changed during stratification, which agrees with the fact that all seeds remained viable throughout the entire period of dormancy. We assume that the release of seed dormancy is not a single-stage process.     

Changes in ascorbate, glutathione, and related enzyme activities during thidiazuron-induced bud break of apple

The changes of ascorbic acid, dehydroascorbic acid, and glutathione content and related enzyme activities were studied in apple buds during dormancy and thidiazuron-induced bud break. An increase in ascorbic acid, reduced form of glutathione (GSH), total glutathione, total non-protein thiol (NPSH) and non-glutathione thiol (RSH) occurred as a result of induction by thidiazuron during bud break, whereas dehydroascorbic acid and oxidized glutathione (GSSG) decreased during the same period. Thidiazuron also enhanced the ratio of GSH/GSSG, and activities of ascorbate free radical reductase (AFR; EC 1.6.5.4), ascorbate peroxidase (EC 1.11.1.11). dehydroascorbate reductase (DHAR; EC 1.8.5.1) and glutathione reductase (GR; EC 1.6.4.2). The ascorbic acid content and the activities of AFR, ascorbate peroxidase, and DHAR peaked when buds were in the side green or green tip stage just prior to the start of rapid expansion, and declined thereafter. The GSH, NPSH, RSH, ratio of GSH/GSSG, and activities of GR increased steadily during bud development.