Seeds of tomato (Lycopersicon esculentum Mill.) which develop in a fully hydrated environment in the fruit switch from a developmental to a germinative mode without a requirement for desiccation

Seed water content is high during early development of tomato seeds (10–30 d after pollination (DAP)), declines at 35 DAP, then increases slightly during fruit ripening (following 50 DAP). The seed does not undergo maturation drying. Protein content during seed development peaks at 35 DAP in the embryo, while in the endosperm it exhibits a triphasic accumulation pattern. Peaks in endosperm protein deposition correspond to changes in endosperm morphology (i.e. formation of the hard endosperm) and are largely the consequence of increases in storage proteins. Storage-protein deposition commences at 20 DAP in the embryo and endosperm; both tissues accumulate identical proteins. Embryo maturation is complete by 40 DAP, when maximum embryo protein content, size and seed dry weight are attained. Seeds are tolerant of premature drying (fast and slow drying) from 40 DAP.Thirty-and 35-DAP seeds when removed from the fruit tissue and imbibed on water, complete germination by 120 h after isolation. Only seeds which have developed to 35 DAP produce viable seedlings. The inability of isolated 30-DAP seed to form viable seedlings appears to be related to a lack of stored nutrients, since the germinability of excised embryos (20 DAP and onwards) placed on Murashige and Skoog (1962, Physiol. Plant. 15, 473–497) medium is high. The switch from a developmental to germinative mode in the excised 30- and 35-DAP imbibed seeds is reflected in the pattern of in-vivo protein synthesis. Developmental and germinative proteins are present in the embryo and endosperm of the 30- and 35-DAP seeds 12 h after their isolation from the fruit. The mature seed (60 DAP) exhibits germinative protein synthesis from the earliest time of imbibition. The fruit environment prevents precocious germination of developing seeds, since the switch from development to germination requires only their removal from the fruit tissue.Abbreviations DAP days after pollination - kDa kilodaltons - SP1-4 storage proteins 1–4 - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - HASI hours after seed isolation - MS medium Murashige and Skoog (1962) medium This work is supported by National Science and Engineering Research Council of Canada grant A2210 to J.D.B.     

Asparaginyl endopeptidase during maturation and germination of durum wheat

Asparaginyl-endopeptidase activity was detected in endosperms of maturing and germinating wheat seeds. The highest activity was found during maturation before the maximal accumulation of storage proteins. The enzyme activity then decreased in the dry seeds and increased again during germination. The increase of activity during germination required the presence of the embryo. In fact, the activity found in detached endosperms was lower than that found in attached ones. The localization at tissue level of the enzyme reveals differences between maturation and germination: the enzyme was about equally located in the aleurone layer and starchy endosperm during maturation, but solely in the aleurone layer during germination. The asparaginyl enzymes from maturing and germinating seeds had many similar properties, such as pH optimum, pH stability, thermal stability and sensitivity to thiol reagents and to thiol compounds. The results suggest that asparaginyl endopeptidases may be involved in the modification of proproteins of storage proteins during seed maturation and in the degradation of storage proteins deposited in the aleurone layer during germination.     

Photoinhibition of radish (Raphanus sativus L) seed germination: control of growth potential by cell-wall yielding in the embryo

The biophysical mechanism underlying photoinhibition of radish (Raphanus sativus L.) seed germination was investigated using three cultivars differing in sensitivity to continuous irradiation with far-red light (high-irradiance reaction of phytochrome). Sensitivity of germination to the inhibitory action of light was assessed by probing germination under osmotic stress (incubation in media of low water potentials adjusted with polyethylene glycol 6000) and expressed in terms of ‘germination potential’ (positive value of the water potential at which germination is inhibited by 50%). Far-red light decreases the germination potential to various degrees in the different cultivars, reflecting the light-sensitivity of germination in water. Removal of the seed coat increases the germination potential by a constant amount in darkness and light. It is concluded that germination depends on the expansive force of the embryo which can be drastically diminished by far-red light. Seed-coat constraint and expansive force of the embryo interact additively on the level of the germination potential. Photoinhibition of germination was accompanied by an inhibition of water uptake into the seed. Analysis of seed water relations showed that osmotic pressure and turgor assumed higher levels in photoinhibited seeds, compared to seeds germinating in darkness, while the water potential was close to zero under both conditions. Far-red light produced a shift (to less negative values) in the curve relating water-uptake rate to external water potential, i.e. a reduction in the driving force for water uptake. It is concluded that photoinhibition of germination results from the maintenance of a high threshold of cell-wall extensibility in the embryo.     

Precociously germinating rapeseed embryos retain characteristics of embryogeny

We compared the germination of Brassica napus L. embryos at three stages of development-mid-cotyledon, maturation and mature dry-to determine at which stage they acquired the capacity for normal germination and seedling development. Embryos were removed from the seed and cultured on hormone-free medium, allowing them to germinate. The transition from embryogeny to germination was monitored both morphologically and biochemically, using synthesis of 12 S storage protein as a marker of embryogeny. The mature embryos (dry seeds) set the standard for normal seedling development: radicle emergence, hypocotyl extension and cotyledon expansion occurred within 2 d and true leaves were formed within a week of germination. Rocket immunoelectrophoresis indicated that the storage proteins in seedlings from mature dry embryos were completely degraded within a week. In contrast, the midcotyledon-stage embryos appeared to germinate abnormally, retaining many embryonic characteristics. Although the roots emerged, the hypocotyls did not elongate and secondary cotyledons instead of leaves were formed at the shoot apex. Also, the seedlings continued to synthesize and accumulate storage proteins. The maturation-stage embryos did develop into normal-looking seedlings, but complete degradation of storage proteins required several weeks, presumably reflecting continued synthesis and turnover. We conclude that embryogenic and germination-specific processes can occur concurrently and that the capacity to develop as normal seedlings is acquired gradually during the maturation process.Abbreviations dpa days post anthesis - EDTA ethylenedi-aminetetraacetic acid - FW fresh weight     

烟草种子成熟过程中膜脂的变化规律研究

该研究采用脂类组学方法,系统地研究了烟草种子成熟过程中膜脂含量及组成比例的变化规律。结果表明:(1)构成叶绿体和类囊体膜的重要脂类质体膜脂的含量及其在总膜脂中的组成比例,在种子成熟的整个过程中保持下降趋势;而构成细胞膜的重要脂类质外体膜脂含量在种子成熟前期则下降显著,在授粉21 d后基本保持不变。(2)总膜脂含量的变化规律与质体膜脂类似,但在授粉后第29天后含量却达到稳定状态。(3)因油脂在种子成熟过程中不断积累,且化学结构与膜脂相似,质体膜脂含量的降低可能与种子成熟过程中种子对油脂累积的持续需求以及对叶绿体及类囊体的需求降低有关。(4)质外体膜脂含量在授粉21 d后基本保持不变的原因,可能是由于脂质外体膜脂是细胞膜组成的主要膜脂,细胞膜在种子成熟以及成熟种子萌发过程中均发挥重要作用,因此质外体膜脂只在种子成熟的前期有部分转化为油脂。     

濒危植物中甸刺玫种子休眠及其生态学意义

对中甸刺玫Rosa praelucens种子结构及其透水性,赤霉素处理对胚的影响,果壳、种皮、胚乳的粗提物活性进行研究。结果表明：(1) 中甸刺玫外种皮是由多层排列紧密的厚壁细胞组成,内种皮为坚硬致密的栅栏组织;(2) 种皮对种子的吸胀阻碍较大, 未处理的种子吸水率较低,吸水13 d后增加量为18.82%;(3) 果实结籽率为0.69%,多数果实中没有饱满的种子;(4) 赤霉素100 mg·kg-1预处理种子可加快胚的萌发速率;(5) 胚乳、种皮、果壳中存在内源抑制物。中甸刺玫种子的休眠是由其形态和生理特点引起的综合休眠。     

小叶兜兰的种子发育和无菌萌发

为建立小叶兜兰的繁育技术体系,本研究通过无菌播种的方法,辅以TTc生活力测定等方法,比较了小叶兜兰种子在授粉后不同发育时期和培养条件下的萌发率,对小叶兜兰种胚的发育过程进行了显微观察,探讨种胚发育程度与萌发的关系。结果表明,小叶兜兰种胚的发育阶段对萌发的影响最大,授粉后255d的种子萌发率最高（90．71％）,该阶段种子仍呈白色但微干燥,种胚刚发育至球形胚阶段,胚柄尚存。1／4MS和1／2MS为小叶兜兰适宜的基本培养基,添加100mg·L^-1的土豆汁对小叶兜兰的无菌萌发有良好的促进作用。     

新疆干旱区植物藜的种子异型性及其萌发机理

新疆干旱区分布的植物藜(Chenopodium album)的种子有黑色和褐色两种类型。对藜的异型性种子从形态结构、不同环境因素及激素或化学物质对萌发的影响以及同工酶谱等方面进行了研究,并对其萌发及适应异质环境的机理进行了讨论。结果表明:(1)藜的异型性种子在形态结构、萌发休眠特性等方面都存在明显差异:黑色种子种皮厚且硬,休眠,萌发慢,萌发率低;褐色种子种皮薄而软,不休眠,萌发快且萌发率高;(2)黑色种子的休眠可通过切除胚根外缘种皮得以完全解除;(3)赤霉素、乙烯利对黑色种子的萌发无明显促进作用;KNO3可较显著促进黑色种子的萌发;协同使用乙烯利和KNO3时,可显著提高黑种子萌发率,完全打破休眠;(4)黑色种子和褐色种子的酯酶、过氧化物酶及过氧化氢酶同工酶谱带存在差异;(5)黑色种子的萌发需要光照,而褐色种子则对光不敏感;低温贮藏对二者的萌发均无显著影响,尽管黑色种子的萌发率有波动。研究结果初步显示黑色种子的休眠是内源(胚)和外源(种皮)因素共同所致。藜的种子异型性及其萌发机理的形成是其对新疆干旱区异质化环境的高度适应。     

Changes in germination and respiratory potential of embryos of dormant Grand Rapids lettuce seeds during long-term imbibed storage,and related changes in the endosperm

Grand Rapids lettuce (Lactuca sativa L.) seeds were stored in an imbibed state for up to two years. Embryos dissected from stored seeds showed a progressive loss with time in their ability to germinate on polyethylene glycol (PEG) solutions. Little germination of dissected embryos from one-month imbibed seeds occurred on-6 bar PEG but only after four months of storage did the dissected embryos fail to germinate on-4 bar PEG. After two years storage 30% of dissected embryos still were able to germinate on-2 bar PEG. This loss of germination potential, which may be a symptom of the development of an embryo dormancy, could be reversed by N⁶-benzyladenine (BA) and red light (R) applied together or separately to dissected embryos. Two weeks of chilling of 12-month imbibed seeds restored sensitivity to R and a 48-h BA pretreatment prior to R resulted in germination rates similar to those of seeds emerging from primary dormancy. There was loss of embryo control of endo--mannanase activity after two weeks of storage even though the endosperms themselves retained their capacity for enzyme synthesis for six more weeks. Eventually, then, endo--mannanase synthesis is not possible because of inherent changes in both the embryo and endosperm, although each tissue undergoes changes at its own rate. Oxygen uptake by embryos dissected from two-month imbibed seeds did not increase to the same extent as embryos dissected from freshly imbibed seeds. In intact seeds germinating from a skotodormant state, oxygen uptake increased at a time coincident with radicle protrusion, but did not achieve the levels of uptake of those seeds germinating from a primary dormant state. The decline in uptake of oxygen by secondary dormant seeds is the result of a lowered respiratory capability of the embryo itself, rather than of changes in permeability of the surrounding structures.Abbreviations BA N⁶-benzyladenine - Pfr active (far-redabsorbing) form of phytochrome - R red light - PEG polyethylene glycol     

In vitro asymbiotic germination of immature seed and formation of protocorm by Cephalanthera falcata (Orchidaceae)

BACKGROUND AND AIMS: Many Orchidaceous species are threatened globally by development and over-collection from their natural habitats for horticultural purposes. Artificial propagation from seeds is difficult in most terrestrial orchids native to temperate regions. Seed production is another limiting factor in the artificial propagation for these species because of the lessened probability of pollination and the destruction of fruit by insect larvae. Members of the genus Cephalanthera are distributed across Europe, Asia and North America. C. falcata is a temperate species of East Asia and an endangered species in Japan. As successful propagation from seeds of this species has never been reported, a reproducible method is described here for seed production in situ and propagation using immature seeds in asymbiotic culture in vitro. METHODS: Effects of hand-pollination and bagging treatment of ovaries were examined. Young capsules were collected every 10 d from 50 d after pollination until 120 d after pollination. Immature seeds obtained from these capsules were cultured asymbiotically on modified Kano medium and ND medium. Seed viability was examined within TTC (2,3,5-triphenyl tetrazolium chloride) test solution and histological observations were made on viable seeds by paraffin embedding at each collection stage. KEY RESULTS AND CONCLUSIONS: Hand-pollination followed by bagging treatment of ovaries with aluminium foil was effective for insect control during fruit development, and successfully yielded capsules. Of the capsules, 74.5 % survived to full maturity. The highest frequency (39.8 %) of seed germination was obtained with seeds harvested 70 d after pollination. The frequency declined with progress of seed maturity on the mother plant. Minimal germination was observed with seeds harvested 100 d or later after pollination. Histological observation suggests that accumulation of such substances as lignin in the inner integument surrounding the embryo during seed maturation plays an important role in induction of dormancy.     

Effects of storage,mucilage presence,photoperiod, thermoperiod and salinity on germination of Farsetia aegyptia Turra (Brassicaceae) seeds: implications for restoration and seed banks in Arabian Desert

Seed viability and germination are key factors in the success of restoration efforts, especially when stored seeds are used. However, the effect of seed storage on germination of most of the native Arabian species is not well documented. We investigated the effect of storage time and role of the seed mucilage in regulating germination, dormancy, salinity tolerance and consequential survival strategy of F. aegyptia in an unpredictable arid desert setting. Effect of light and temperature during germination was studied under two photoperiods and two thermoperiods using intact and de-mucilaged seeds. Presence of mucilage and thermoperiod did not affect the germination. However, seed collection year and photoperiod had a highly significant effect on the germination. Increasing salinity levels decreased the germination of F. aegyptia but ungerminated seeds were able to germinate when salinity stress was alleviated. Seed storage at room temperature enhances the germination percentage, indicating that F. aegyptia seeds have physiological dormancy and it can be alleviated by after-ripening at dry storage. In addition, F. aegyptia seeds show ability to germinate at lower salinity concentration and remain viable even at higher saline conditions, indicating their adaptability to cope with such harsh environmental conditions.     

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 ).     

红皮糙果茶种子生活力测定与发芽试验

选取饱满有光泽的红皮糙果茶Camellia crapnelliana种子,进行种子贮存生活力测定、种子发芽试验及种皮解剖结构观察等研究,就种子繁殖探讨其濒危原因。结果表明,有利于保持种子生活力的贮存方式为：4 ℃贮存>常温贮存>－10 ℃贮存;种子发芽试验中,采用不同浓度赤霉素(GA₃)、吲哚丁酸(IBA)和不同温度温水处理,种子发芽率均不高,最高为40%;不同因素对种子发芽的影响不同,种子发芽最佳处理为800 mg·L^-1GA₃浸泡6 h;种皮解剖结构观察到红皮糙果茶种皮坚硬,由栓化壁和骨状石细胞组成,内、外部石细胞垂直交错排列,这可能是造成种子萌发困难的机械阻碍原因。     

A threatened alpine species,Fritillaria tubiformis subsp. moggridgei: Seed morphology and temperature regulation of embryo growth

Abstract

Alpine plants have evolved to fit their life cycle into the short vegetative season of mountain habitats. Fritillaria tubiformis Gren. & Godr. subsp. moggridgei (Boiss. & Reuter ex Planch.) Rix (Liliaceae) is an endemic alpine geophyte, bearing seeds with underdeveloped embryos. Seeds are dispersed in August and embryos complete their development by spring when seeds germinate. In order to optimize seed banking procedures and to develop a proper germination protocol for plant regeneration, we studied embryo morphogenesis and analyzed how this process is influenced by temperature. Radicle protrusion occurred after an incubation of 5 months at 4°C. Under these conditions, underdeveloped embryos reached maturity and acquired a well-defined shoot apex. At the time of dispersal, abundant storage compounds were present in seeds. Lipids and lipid/proteins were uniformly distributed within the embryo and the endosperm, respectively. At late stages of embryo development, starch granules were localized at the cotyledonary tip and were also detected around the shoot meristem. Results suggested that F. tubiformis embryos resumed growth over a large range of temperatures, but were only able to complete development at low temperatures after which they were able to germinate by spring.     

Changes in the amounts of chlorogenic and caffeic acids during tobacco seed maturation

During maturation of tobacco seed, the amount of chlorogenic acid rapidly increased between days 7 and 11 after anthesis and then decreased until day 15. Chlorogenic acid was not detected in fully mature seed. On the other hand, caffeic acid appeared from day 15, reached a maximum at day 21, and rapidly disappeared by day 25 although the amount was always low in comparison with that of chlorogenic acid. Changes in dry wt, water content, testa color and germination of tobacco seeds during maturation were also investigated. The results suggested that there were rapid biochemical changes in seed between days 11 and 15 after anthesis.     

Endo-β-mannanase activity during dormancy alleviation and germination of white spruce (Picea glauca) seeds

It is not known how embryos of seeds of the Pinaceae protrude from their enclosing tissues to complete germination. Prior to protrusion of the radicle there is an increase in endo-β-1,4-mannanase (EC 3.2.1.78) activity associated with weakening of the micropylar megagametophyte/nucellus from seeds of white spruce ( Picea glauca [Moench.] Voss). Mannanase activity is present as three isoforms (pI values 5.0, 4.8, 4.7) in both the embryo and surrounding structures (megagametophyte and nucellus) prior to and during imbibition. Activity of all the isoforms increases in the chalazal and micropylar megagametophyte during germination. Activity then declines after the testa splits, typically 1 day prior to radicle protrusion, due partially to its leaching from the seed into the surrounding water. Activity increases in the cotyledons and axis as the embryo commences elongation. Seeds from dormant seedlots exhibit a lower germination percentage, relative to seeds from nondormant seedlots, and the force necessary for the embryo to puncture the surrounding structures tends to be greater. Although similar mannanase activities are present in unimbibed seeds of dormant and nondormant seedlots, during germination, enzyme activity in seeds of dormant seedlots is lower. Moist chilling alleviates dormancy in the seeds of the Pinaceae and, during 3 weeks of this treatment, mannanase activity slowly increases. After 3 weeks of moist chilling and regardless of whether the seedlot was dormant or not prior to moist chilling, the force necessary to puncture the micropylar megagametophyte and nucellus is lower, and the speed of germination greater. Seeds from previously dormant seedlots also complete germination to a greater percentage, relative to unchilled seeds from dormant seedlots. Upon transfer to 25°C, mannanase activity in moist-chilled seeds decreases during germination of all seedlots regardless of their previous dormancy status.     

Seed germination requirements of Trembleya laniflora (Melastomataceae), an endemic species from neotropical montane rocky savannas

Trembleya laniflora is an endemic shrub from neotropical montane rocky savannas of southeastern Brazil. It has been indicated as a potential candidate for ecological restoration of abandoned iron‐ore mines due to heavy metal accumulation. Here, we evaluated the seed germination requirements of T. laniflora. Seeds collected in 2005 and 2008 were set to germinate under a broad range of temperature and light conditions. Seed viability was estimated by dissecting seeds under a dissecting microscope for embryo presence/absence. Seeds were photoblastic and optimum temperature range was 20–25°C, coinciding with the onset of the rainy season. Seeds were viable after 42 months of storage, which together with small seeds that easily get buried and light requirement for germination suggest formation of soil seed banks. Except the large fraction of embryoless seeds, almost all tested seeds germinated when incubated under light conditions; therefore, T. laniflora should be regarded to have nondormant seeds. Easiness of burial resulting from small seed size and positive photoblastism may both contribute to incorporation into soil seed banks. Our data suggest that the long‐term storage of T. laniflora seeds provides a useful strategy for plant reintroduction.     

度胁迫对罂粟种子萌发影响及其活力变化规律的研究

为妥善保存罂粟种质资源及防止种子老化,研究了温度对罂粟种子萌发及种子活力的影响。结果表明:罂粟为高温敏感低温萌发型种子,发芽的适宜温度为10～20℃,最适温度为18℃左右,低于5℃和高于20℃的温度,种子萌发受到抑制;在对其进行丸化包衣时,加热温度应控制在60℃以内,时间应控制在60min以内;在常温库存条件下,罂粟种子活力随贮藏年限的增加而下降,贮藏时间超过五年后,发芽率低于50%,不宜作种用,在作为种质资源保存时,须每隔五年繁殖一次。     

The presence of a p53-like protein during pea seed maturation and germination

ABSTRACT

The mechanisms that allow monitoring of DNA damage and the activation of repair systems in plants are poorly known. In mammalian cells the tumor suppressor protein p53 plays an important role in the checkpoint pathway induced by DNA damage. In this work, we investigated the presence and distribution of the p53-like protein in pea root tip nuclei and its role during early germination in relation to DNA damage. In pea seed, PFGE and TdT assays show that DNA fragmentation occurs during maturation and dry seed storage, and that this DNA fragmentation is repaired at the beginning of germination before the onset of proliferation. In the same seeds, the p53-like protein was found during maturation and germination. Immunoblotting characterization of this protein led to the identification of a single specific protein of about 94 kDa, more abundant at the beginning of the hydration process than in actively cycling cells. Furthermore, the p53-like protein revealed different nuclear distribution patterns, probably in relation to the formation of DNA fragments in dry seeds, and to the reactivation of repair mechanisms during early germination. These data suggest that the presence of a p53-like protein in quiescent or proliferating pea embryo cells is related to DNA damage, and serves for the maintenance of genetic information and the development of normal seedlings.     

脱落酸调控种子休眠和萌发的分子机制

脱落酸(ABA)是调控种子休眠和萌发过程的主要植物激素。种子内源ABA含量和种胚对ABA敏感性共同调控种子休眠和萌发过程, 确保植物种子以休眠状态在逆境中保持其自身繁衍能力, 并在适宜的环境下启动萌发程序。种子ABA合成代谢和ABA信号转导途径涉及许多重要基因家族, 它们通过复杂的调控网络精确地控制着种胚发生、种子成熟、休眠及萌发进程。该文对ABA调控种子休眠和萌发的分子机制最新研究进展进行综述, 并展望了今后的研究方向。