桃儿七种子低温层积过程中胚形态及生理生化物质变化

为探究低温层积过程中桃儿七种子胚形态及生理生化变化与休眠解除的内在联系,该研究通过低温层积处理(90 d)解除桃儿七种子休眠,观测不同层积时间种子胚形态、胚率、发芽率、营养物质(淀粉、可溶性蛋白质、可溶性糖)含量、内源激素［赤霉素(GA)、吲哚乙酸(IAA)、脱落酸(ABA)］水平及呼吸途径关键限速酶［丙酮酸激酶(PK)、琥珀酸脱氢酶(SDH)、6 磷酸 葡萄糖脱氢酶(G 6 PDH)］的活性变化。结果显示：(1)在低温层积过程中,桃儿七种子胚形态为鱼雷或子叶型胚;种子发芽率在层积后期(60~75 d)显著提高(P＜0.05)。(2)层积后,种子内淀粉含量及PK活性、SDH活性显著降低(P＜0.05),其可溶性蛋白含量和IAA含量显著升高(P＜0.05),萌发促进物和抑制物比例(GA/ABA、IAA/ABA、GA+IAA/ABA)也呈升高趋势。(3)种子胚率与其可溶性糖含量呈显著负相关关系,种子发芽率与其可溶性蛋白呈显著正相关关系(P＜0.05)。研究发现,桃儿七种子无形态休眠;种子内营养物质的分解转化为种子休眠解除过程中各种代谢活动提供能量,且淀粉可能是此过程中最主要的供能物质;磷酸戊糖途径(PPP)的活化、萌发促进物和抑制物比例的升高及IAA含量的显著上升是桃儿七解除休眠的关键。     

桃儿七种子解剖结构及其萌发生长期形态特征

为探明种皮和胚乳是否是限制桃儿七种子萌发的主要因素,利用组织切片和显微技术,对桃儿七种子及其不同萌发期（1、7、14、21、28 d）解剖结构和播种后一定时期内（7~210 d）的植株生长形态进行观察。桃儿七种子由种皮、胚乳和胚构成。种皮包括外种皮和内种皮,外种皮致密规整,由外至内分别为栅状石细胞和表皮层细胞,内种皮由5~6层海绵细胞组成。胚乳占种子体积的绝大部分,包括珠孔胚乳和外胚乳。胚由胚根、胚轴和子叶组成,被致密种皮、多层珠孔胚乳和外胚乳包围。萌发期1~7 d胚根和胚轴开始伸长,7~14 d两片子叶分离,14~21 d胚根突破珠孔胚乳和种皮,21~28 d胚根、胚轴和子叶继续扩张伸长。种子播种210 d后可平均形成3片功能真叶和5条不定根。致密种皮（物理休眠）和多层胚乳（机械休眠）是限制桃儿七种子萌发的两个主要因素。     

濒危植物桃儿七种子休眠特性的研究

桃儿七种子在自然条件下具有休眠期长、萌发不良的生理特性,为了探讨和研究桃儿七种子的休眠特性,利用分离胚培养、生物鉴定法、GA₃浸种、以及综合利用GA₃和低温层积处理等方法。结果表明：种皮和胚乳的限制以及生理后熟是引起桃儿七种子休眠的主要原因,用400 mg·L^-1的GA₃溶液浸种24 h或低温层积后用GA₃处理均能在一定程度上解除休眠促进萌发,其中以低温层积90 d后用500 mg·L^-1的GA₃浸种36 h效果最好,发芽率和发芽势分别达到81.11%和50.00%。     

珍稀濒危植物金丝李种子的休眠机理

珍稀濒危植物金丝李(Garcinia paucinervis)种子的萌发十分缓慢,探讨其休眠机理,可为该物种的种质资源保育与可持续利用提供理论依据。本文对金丝李种子种皮结构及其透水性,剔除部分种皮和胚乳后种子的萌发情况,胚乳和胚等粗提物的活性,储藏、层积和不同温度下种子萌发情况,萌发过程中内源激素含量等进行了研究。结果表明:金丝李种皮无栅栏细胞层,下表面的角质层较薄;种皮对种子的吸胀阻碍小;随着种孔端剔除种皮和胚乳程度的加深,金丝李种子的萌发进程逐渐延长,甚至降低其萌发率,种脐端削除处理对种子萌发影响不大;内果皮、种皮、胚乳和胚中可能存在抑制金丝李种子萌发和生长的内源抑制物;新鲜种子胚率达86.12%,低温层积后胚率无显著变化;低温层积处理延缓其萌发进程,对萌发率无显著影响,4℃低温层积是储藏金丝李种子的较好方法;种子萌发对温度敏感,在32℃培养下可打破种子休眠,萌发速度显著加快。种子萌发过程中ABA含量降低,GA与ABA、IAA与ABA的比值随种子萌发显著升高,萌发促进与抑制物比例逐渐趋于提高。因此,金丝李种子存在内源抑制物,同时缺乏萌发促进物质,导致生理休眠。该种子休眠特性使其幼苗生长能应对生境的季节变化,种群在风险环境中得以延续,避免大量幼苗竞争。植被破坏导致种子萌发阶段受阻是造成金丝李濒危的原因之一。     

影响琼花种子休眠的因素

琼花种皮有吸水性,休眠非因种皮的不透水性造成.种仁中存在导致休眠的抑制物质,胚的抑制物质含量最高.种子须先经4℃低温层积60 d,再经25℃暖温处理90 d,而后在较低温(15℃)条件下才能解除休眠而萌发.当年成熟种胚在翌年10月才能彻底破除休眠.6-BA和GA对种胚破眠均无明显作用.     

裕民贝母种子形态休眠解除过程中的形态学研究

为掌握裕民贝母种子休眠的形态响应机制,判断外源GA_3能否有效解除种子形态休眠,对CK和GA_3:30 mg/L处理下种子的形态结构及亚微结构变化规律进行比较。结果表明:(1)GA_3:30 mg/L处理下种子的形态休眠时间为60 d,比CK处理提前20 d,胚率从22.33%提高到87.67%,然后种子进入生理休眠阶段;(2)种子具翅,质轻型小,胚完成形态休眠后结构分化仍不明显;(3)随种子形态休眠的打破,种皮颜色不断加深,胚乳不断水解,胚不断生长发育,胚率提高至恒定;(4)随种子形态休眠的解除,种皮气孔器结构稳定,但叶绿体数量持续增加,种皮、胚乳及胚的亚微结构变化不明显,仅胚乳表皮细胞破损严重。     

东京野茉莉种子休眠机制及其破除方法初探

以采自江西吉安官山林场5年生东京野茉莉当年自然带壳种子为材料,通过对其种子吸水率、不同层积时期种子内生理生化指标的测定、种子萌发抑制物分析,并利用各类不同药剂处理进行发芽试验,以探寻东京野茉莉种子的休眠机理及破眠方法。结果表明:(1)休眠的原因主要包括种皮障碍、缺少萌发所需激素以及种皮、胚中存在萌发抑制物,其中种皮障碍和抑制物的存在是限制种子萌发的首要因素。(2)GA3处理结合自然低温层积30d即可解除东京野茉莉种子胚的休眠,但种皮障碍始终是其种子萌发的限制因素。(3)GA3、NAA、6-BA等药剂处理均可促进种子的萌发,并以刻伤种子后用500mg/L GA3处理24h为破除该种子休眠的最有效方法。     

离体条件下TDZ对解除北五味子种子休眠的影响

以深度休眠的北五味子种子为材料,研究了离体条件下,种皮、胚乳因素对北五味子种子休眠的影响;建立了离体条件下,使用TDZ有效解除北五味子种子休眠的方法;并对北五味子种子解除休眠过程中过氧化物酶和过氧化氢酶活性变化规律进行分析.结果表明,(1)种皮和胚乳因素是引起北五味子种子休眠的主要因素,种皮和胚乳对北五味子种子的萌发具有不同程度的抑制作用;(2)MS培养基中添加0.02 mg·L-1 TDZ能有效解除北五味子种子休眠,可使第30天的发芽率达到90.0%,显著缩短北五味子种子休眠时间;(3)随着北五味子种子休眠被解除,过氧化物酶和过氧化氢酶活性均呈明显的增加趋势.     

绞股蓝种子休眠机理及其破除方法研究

以采自广西金秀县的绞股蓝种子为研究材料,对其休眠原因、休眠类型及其破眠方法进行了研究,为绞股蓝种子繁殖提供理论依据和技术支持。结果表明:(1)绞股蓝新采收成熟种子的生活力达91%,在10℃~35℃恒温和15℃/25℃变温中的发芽率均低于10%,新种子的生活力极显著大于发芽率,具有显著的休眠现象。(2)绞股蓝种皮不限制吸水,胚分化发育完全,离体胚发芽率为(78.0±4.8)%,且能够长成正常幼苗,说明绞股蓝种子的胚在离体条件下无休眠现象。(3)绞股蓝完整种子及其粉碎种子的水提液对白菜种子的萌发率、苗高及根长均有抑制作用,随水提液浓度增加抑制作用均显著增强,且粉碎种子的抑制作用较强;当粉碎种子的水提液浓度为5%时白菜种子萌发率、苗高、根长分别为18.0%、0.1cm、0.1cm,分别显著低于对照77.1%、97.3%、95.8%,说明绞股蓝种子的种皮和胚乳中存在水溶性萌发抑制物质,是绞股蓝种子休眠的主要原因。(4)GA3和6-BA不能促进绞股蓝种子萌发,低温层积对绞股蓝种子休眠的解除具有促进作用;绞股蓝种子的休眠属于生理休眠类型,休眠水平属于中间型。(5)低温干藏能够打破绞股蓝种子休眠,是绞股蓝种子破除休眠及种子保存较为理想的方式。     

东北刺人参种子萌发影响因子的研究

东北刺人参(Oplopanax elatus Nakai)种子透水性良好,休眠后萌发不受其影响。种皮和胚乳的水提取物中存在萌发抑制物质,胚乳中提取物对白菜种子萌发的抑制效果比种皮更明显。种子自然脱落时胚尚未分化完全,处于心形胚阶段。种子需要先温暖层积以完成胚的分化与生长,然后转入低温层积完成生理后熟。同批种子胚的发育不完全同步,变温层积处理7个月有极少数种子萌发,连续变温层积处理17个月大部分种子萌发。不同年份受气候条件影响,种子产量和发芽率差异较大。种子耐贮性较强,贮藏2年的种子生活力变化不大,仍具有较高的萌发潜力。     

A Seed Coat Bedding Assay to Genetically Explore In Vitro How the Endosperm Controls Seed Germination in Arabidopsis thaliana

The Arabidopsis endosperm consists of a single cell layer surrounding the mature embryo and playing an essential role to prevent the germination of dormant seeds or that of nondormant seeds irradiated by a far red (FR) light pulse. In order to further gain insight into the molecular genetic mechanisms underlying the germination repressive activity exerted by the endosperm, a "seed coat bedding" assay (SCBA) was devised. The SCBA is a dissection procedure physically separating seed coats and embryos from seeds, which allows monitoring the growth of embryos on an underlying layer of seed coats. Remarkably, the SCBA reconstitutes the germination repressive activities of the seed coat in the context of seed dormancy and FR-dependent control of seed germination. Since the SCBA allows the combinatorial use of dormant, nondormant and genetically modified seed coat and embryonic materials, the genetic pathways controlling germination and specifically operating in the endosperm and embryo can be dissected. Here we detail the procedure to assemble a SCBA.     

The mericarp of the halophyte <Emphasis Type="Italic">Crithmum maritimum</Emphasis> (Apiaceae): structural features,germination, and salt distribution

At maturation and during seed fall and dispersal, halophyte seeds may be subjected to invasion by salt ions. How these seeds remain viable in such hostile environments is however still unclear, depending for instance on the species and the family. In the Apiaceae, the mericarp (fruit) shows a wide range of morphological and anatomical modifications, many of which may enhance the adaptation to severe environmental conditions. Therefore, structural features, ion accumulation, and long-term floating capacity were investigated in the fruit (mericarp) of the halophyte Crithmum maritimum L. The mericarp was composed of a spongy outer coat, a secretory envelope, a thin endocarp reduced to a unicellular layer delimiting the endosperm, and an embryo. Both of the secretory canals and the endocarp adhered after complete ripening of the mericarp, while the epicarp and much of the mesocarp formed the spongy coat. Assessing long-term floating ability of the fruit under laboratory conditions revealed that even after 60 d, more than 98% of C. maritimum L. mericarps still floated over seawater. Seed germination was delayed and reduced by the spongy coat. The X-ray microanalysis revealed that the spongy coat and the secretory canals contained essentially Cl and Na, while seeds, i.e. endosperm and embryo, accumulated mostly Mg, K and P. In a subsequent experiment designed to simulate salt leaching by rain, most of the salt accumulated in the spongy coat and seeds was released after 2 h imbibition in distilled water. Taken together, these results highlight the protective role of the mericarp and the likely involvement of this structure in the seed dispersal of C. maritimum L. This may ultimately have eco-physiological implications explaining the successful establishment of this halophyte in its native saline biotopes.     

Seed structure and germination in Cuscuta pedicellata with some notes on C. campestris

Dry seeds of Cuscuta pedicellata have a deeply pitted surface due to invaginated epidermal cell walls. After water uptake these walls bulge outwards and the seed surface becomes papillose. The seed coat consists of an epidermis, two palissade cell layers, and a multiple layer of parenchyma cells. The epidermis contains starch and mucilage, the parenchyma cells are compressed but some contain starch. The endosperm consists of starch–filled cells, but has a peripheral aleuron layer. The endosperm cell walls are gelatinous. The variable structure of the seed coat epidermis is believed to function in wind dispersal and rapid water uptake. Seed dormancy is common in the genus, but does apparently not occur in C. pedicellata.     

Seed development ofAbelmoschus esculentus (Malvaceae)

The endosperm is nuclear, cell wall initiation starts 5 days after pollination. During early stages endosperm nuclei exhibit synchrony in their division. Embryogeny is of the Asterad type. A7-to 10-celled suspensor persists up to the dicot stage of the embryo. Both integuments contribute towards formation of the seed coat. 30 days after pollination seeds become mature. Their endosperm is scanty and persists as a thin layer between the folds of the cotyledons. Nucellus remnants are present towards the funicular side.     

Degree of pectin methyl esterification in endosperm cell walls is involved in embryo bending in Arabidopsis thaliana

The endosperm is a transitory structure involved in proper embryo elongation. The cell walls of mature seed endosperm are generally composed of a uniform distribution of cellulose, unesterified homogalacturonans, and arabinans. Recent studies suggest that changes in cell wall properties during endosperm development could be related to embryo growth. The degree of methyl esterification of homogalacturonans may be involved in this endosperm tissue remodelling. The relevance of the degree of homogalacturonan methyl esterification during seed development was determined by immunohistochemical analyses using a panel of probes with specificity for homogalaturonans with different degrees of methyl esterification. Low-esterified and un-esterified homogalacturonans were abundant in endosperm cells during embryo bending and were also detected in mature embryos. BIDXII (BDX) could be involved in seed development, because bdx-1 mutants had misshapen embryos. The methyl esterification pattern described for WT seeds was different during bdx-1 seed development; un-esterified homogalacturonans were scarcely present in the cell walls of endosperm in bending embryos and mature seeds. Our results suggested that the degree of methyl esterification of homogalacturonans in the endosperm cell wall may be involved in proper embryo development.     

Olive embryo in vitro germination potential: role of explant configuration and embryo structure among cultivars

The in vitro germination of excised embryos can break dormancy rapidly and shorten the time required to produce seedlings, speeding up olive breeding programmes as well as rootstock production. In this study, the in vitro germination potential of four Sicilian olive cultivars was evaluated during two years of experiments, using explants with three different morphological configurations that represent three different degrees of embryo exposure: (1) intact stoneless seeds containing the embryo, the endosperm and the seed coat (Emb+En+SC), (2) seeds without the seed coat (Emb+En) and (3) naked, isolated embryos (seed coat and endosperm both removed: Emb). Differences were found in the germination percentages and timing due to both genotype and explant type. The root and shoot meristems, the radicle-hypocotyl axis, the provascular tissues and embryo storage reserves were identified as embryo anatomical structures which could influence germination capacity. Observation of these structures, however, indicated similar germination potential among cultivars, suggesting possible differences in other dormancy factors. In spite of variation in cultivar performance, after 60 days of in vitro culture all cultivars demonstrated the highest germination of naked embryos (explant type 3) and lowest for stoneless seeds (explant type 1); stoneless seeds without the seedcoat (explant type 2) showed intermediate germination percentages.     

Germination ecophysiology of Annona crassiflora seeds

BACKGROUND AND AIMS: Little is known about environmental factors that break morphophysiological dormancy in seeds of the Annonaceae and the mechanisms involved. The aim of this study was to characterize the morphological and physiological components of dormancy of Annona crassiflora, a tree species native to the Cerrado of Brazil, in an ecophysiological context. METHODS: Morphological and biochemical characteristics of both embryo and endosperm were monitored during dormancy break and germination at field conditions. Seeds were buried in the field and exhumed monthly for 2 years. Germination, embryo length and endosperm digestion, with endo-beta-mannanase activity as a marker, were measured in exhumed seeds, and scanning electron microscopy was used to detect cell division. The effect of constant low and high temperatures and exogenous gibberellins on dormancy break and germination was also tested under laboratory conditions. KEY RESULTS: After burial in April, A. crassiflora seeds lost their physiological dormancy in the winter months with lowest monthly average minimum temperatures (May-August) prior to the first rainfall of the wet season. The loss of physiological dormancy enabled initiation of embryo growth within the seed during the first 2 months of the rainy season (September-October), resulting in a germination peak in November. Embryo growth occurred mainly through cell expansion but some dividing cells were also observed. Endosperm digestion started at the micropylar side around the embryo and diffused to the rest of the endosperm. Exogenous gibberellins induced both embryo growth and endo-beta-mannanase activity in dormant seeds. CONCLUSIONS: The physiological dormancy component is broken by low temperature and/or temperature fluctuations preceding the rainy season. Subsequent embryo growth and digestion of the endosperm are both likely to be controlled by gibberellins synthesized during the breaking of physiological dormancy. Radicle protrusion thus occurred at the beginning of the rainy season, thereby maximizing the opportunity for seedlings to emerge and establish.     

槐种子发育中胚乳细胞半乳甘露聚糖积累的研究

槐 ( Sophora japonica L.)开花约 60 d至种子成熟 ,为胚乳半乳甘露聚糖积累期。用组织化学方法 ,对储藏于胚乳细胞壁上的半乳甘露聚糖的形成积累进行了观察 ,结果表明 ,半乳甘露聚糖最先在邻近胚的胚乳细胞的粗面内质网的囊泡腔内形成 ,并通过细胞质膜分泌至细胞壁周围。此后 ,半乳甘露聚糖的积累逐渐向种皮方向扩展 ,及至种子成熟时 ,除糊粉层外 ,所有胚乳细胞几乎全由多糖所填充。此外 ,对半乳甘露聚糖发生部位及其积累过程的消长变化进行了讨论     

Structure and Histochemistry of Embryo Envelope Tissues in the Mature Dry Seed and Early Germination of Phacelia tanacetifolia

The embryo envelope tissues in both mature dry seed and duringearly germination of Phacelia tanacetifolia were investigatedby bright-field and fluorescence light microscopy and scanningelectron microscopy. The ruminate seed had an irregularly reticulatesurface owing to the presence of polygonal areas, correspondingto the cells of the seed coat. The raised margins of these cellsjoined at the lobe tips, where radially arranged thickeningsoccurred. The unitegmic seed coat was made up of three distinctlayers: the frayed outer layer, the middle layer with portionsrising outwards to form the radial thickenings, and the innerlayer, the thickness of which was greatest in the micropylarzone. The endosperm tissue had two regions, the micropylar andthe lateral endosperm, which differed in polysaccharide composition,thickness and metachromasy intensity, and presence (in the lateralendosperm) or absence (in the micropylar endosperm) of birefringenceof the cell walls. Moreover, in the micropylar region, wherethe embryo suspensor remnant was found, Ca-oxalate crystalswere scarce or absent. The presence of a partially permeablecuticle covering the seed endosperm was observed. Incubationof seeds in Lucifer Yellow CH indicated that water was ableto penetrate quickly into the seed coat along the pathway formedby the radial thickenings, the raised margins of the polygonalcells and the middle layer. Afterwards, LY-CH readily infiltratedthe apical portions of the seed lobes and then the whole endosperm.Following imbibition, morphological changes were found in themicropylar endosperm, such as the initial digestion of proteinbodies. In addition, both in the seed coat and in the endosperm,a weaker fluorescence, probably due to leaching of polyphenolicsubstances, was observed. Once the seed coat was broken at themicropylar end of the seed, the endosperm cap surrounding theradicle tip had to be punctured by it so that complete germinationcould occur. Weakening and rupture of the micropylar endospermare briefly discussed. Copyright 2000 Annals of Botany Company Phacelia tanacetifolia, seed coat, micropylar endosperm, endosperm cap, early germination, structure, histochemistry     

Seed coat development, anatomy and scanning electron microscopy of Harpagophytum procumbens (Devil's Claw), Pedaliaceae

Seed coat development of Harpagophytum procumbens (Devil's Claw) and the possible role of the mature seed coat in seed dormancy were studied by light microscopy (LM), transmission electron microscopy (TEM) and environmental scanning electron microscopy (ESEM). Very young ovules of H. procumbens have a single thick integument consisting of densely packed thin-walled parenchyma cells that are uniform in shape and size. During later developmental stages the parenchyma cells differentiate into 4 different zones. Zone 1 is the multi-layered inner epidermis of the single integument that eventually develops into a tough impenetrable covering that tightly encloses the embryo. The inner epidermis is delineated on the inside by a few layers of collapsed remnant endosperm cell wall layers and on the outside by remnant cell wall layers of zone 2, also called the middle layer. Together with the inner epidermis these remnant cell wall layers from collapsed cells may contribute towards seed coat impermeability. Zone 2 underneath the inner epidermis consists of large thin-walled parenchyma cells. Zone 3 is the sub-epidermal layers underneath the outer epidermis referred to as a hypodermis and zone 4 is the single outer seed coat epidermal layer. Both zones 3 and 4 develop unusual secondary wall thickenings. The primary cell walls of the outer epidermis and hypodermis disintegrated during the final stages of seed maturation, leaving only a scaffold of these secondary cell wall thickenings. In the mature seed coat the outer fibrillar seed coat consists of the outer epidermis and hypodermis and separates easily to reveal the dense, smooth inner epidermis of the seed coat. Outer epidermal and hypodermal wall thickenings develop over primary pit fields and arise from the deposition of secondary cell wall material in the form of alternative electron dense and electron lucent layers. ESEM studies showed that the outer epidermal and hypodermal seed coat layers are exceptionally hygroscopic. At 100% relative humidity within the ESEM chamber, drops of water readily condense on the seed surface and react in various ways with the seed coat components, resulting in the swelling and expansion of the wall thickenings. The flexible fibrous outer seed coat epidermis and hypodermis may enhance soil seed contact and retention of water, while the inner seed coat epidermis maintains structural and perhaps chemical seed dormancy due to the possible presence of inhibitors.