A new anti-counterfeiting method: fluorescent labeling by safranine T in tobacco seed

Counterfeit seeds cause a large loss in seed industry and crop production. Present study was undertaken to find a new anti-counterfeiting technology by labeling seeds with a fluorescent compound, safranine T (ST). Response surface analysis (RSA) experiment with two factors (ST concentration and seed soaking time) of 14 combinations was performed to investigate the ST effects on seed vigor and fluorescence showing in seedlings of two tobacco varieties, MS Yunyan 85 and Honghua Dajinyuan. After soaking in ST solutions, tobacco seeds were germinated for 16 days, then the germination index (GI), vigor index (VI), length of roots and shoots were determined. The optimum combinations of ST concentration and soaking time on GI, VI and length of shoots and roots were 0.57–0.69 mg/ml and 6.77–9.34 h for MS Yunyan 85 and 0.51–0.80 mg/ml and 5.81–7.39 h for Honghua Dajinyuan, respectively. Meanwhile, tobacco seedlings, treated with ST solutions from 0.3 to 1.5 mg/ml showed obviously fluorescence under illumination of green light. It suggests that soaking tobacco seeds in safranine T solution of suitable concentration can be used as a labeling technique for anti-counterfeiting in tobacco seeds.     

Chilling tolerance in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> induced by seed priming with putrescine

Chilling stress is one of primary constraints to tobacco production in many parts of the world. The present study was conducted to induce chilling tolerance in tobacco by seed priming with putrescine (Put) in relation to physiological changes, using seeds from two tobacco varieties, MSk326 (chilling sensitive variety) and Honghuadajinyuan (HHDJY, chilling tolerant variety). Seed germination, seedling antioxidant enzyme activities and malondialdehyde (MDA) concentration, as well as polyamine concentration were determined under low temperature. During chilling stress at 11°C, seed priming with 0.01 mM Put for 48 h (Put0.01mM48 h) and seed priming with 0.1 mM Put for 48 h (Put0.1mM48 h) significantly increased germination percentage, germination index, seedling length and dry weight of both varieties compared to the controls without Put treatment. When seedlings of 4-leaf stage suffered a short chilling stress (5°C), Put 0.1 mM 48 h improved the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), increased endogenous Put, Spd and Spm concentration and decreased the MDA concentration. The results showed that Put priming treatments were available to enhance the chilling tolerance of tobacco seedlings. The optimal treatment of Put was Put0.1 mM48 h.     

六盘山区辽东栎的实生苗更新及其影响因子

研究了六盘山区辽东栎(Quercus liaotungensis)灌丛的种子产量、土壤种子库组成、湿沙和风干贮藏对种子寿命的影响以及动物取食子叶对种子萌发和幼苗建立的影响。结果表明, 辽东栎灌丛种子的完好率为27.51%, 被动物取食或搬运种子的比例(41.51%)显著高于其他类型种子(p < 0.01); 辽东栎次生林土壤种子库中萌发和虫蛀种子分别占35.16%和38.29%, 完好种子仅占13.65%, 捕食动物主要通过贮藏或搬运而影响土壤种子库中的种子密度。湿沙贮藏60天的辽东栎种子自动萌发率高达96.67%, 短期贮藏可加快种子的萌发进程, 提高萌发率和萌发指数, 但随着贮藏时间的延长, 种子萌发进程延迟, 萌发率、萌发指数和活力指数均不同程度地降低。排除动物取食处理的幼苗在林窗和林下生境的存活率分别为80%和83%, 而不排除动物取食幼苗在2种生境中分别仅有25%和31%能够存活, 表明子叶在幼苗建立中具有重要作用。林窗中幼苗子叶的动物取食率(85.00%)高于林下(71.00%), 子叶留存的幼苗在林窗中的存活率(6.00%)低于林下(15.50%), 而子叶被取食幼苗的存活率在两种环境中基本相等(分别为18.50%和18.00%)。     

Review of seed pelletizing strategies for arid land restoration

Arid lands face numerous restoration challenges due to infrequent and irregular precipitation which impacts plant germination, growth, and survival. Abundant seed predators, harsh surface conditions, and native seeds often poorly suited to mechanical distribution compound restoration challenges. Seed pellets (a.k.a. pods, seed balls, and seed bombs), an aggregation of clay, soil, water, and multiple seeds, have the potential to reduce some of the challenges. However, no formal guidelines based on an aggregated research review exist. Available publications, both peer‐reviewed and any public domain, were reviewed. Of the 24 publications found (some describing multiple tests), seed pellets tested on rangelands had 9 negative effects, 10 neutral effects, and 8 positive effects. Greenhouse testing showed five negative effects, two neutral effects, and four positive effects. Forest systems had three neutral effects and two positive effects. Advantages in mechanical distribution were not well quantified and other effects were lumped together in germination and seedling establishment totals. The use of pelleted seed would benefit from a more mechanistic understanding of the interaction between the pellets and the systemic constraints. As long as the pellets themselves do not greatly reduce germination or establishment, they appear a potentially viable method for improving seed distribution and seeding efficiency, protecting seeds, and adding amendments. While the evidence is far from conclusive, seed pellets may also hold some advantages in increasing germination and establishment.     

Efficiency of endozoochorous seed dispersal in six dry-fruited species (Cistaceae): from seed ingestion to early seedling establishment

We combined laboratory and nursery experiments to analyse the effectiveness of sheep as endozoochorous seed dispersers of six native shrubby Cistaceae species collected in SE Spain (Helianthemum apenninum (L.) Mill., H. violaceum (Cav.) Pers., Fumana ericoides (Cav.) Grand., F. thymifolia (L.) Spach, Cistus monspeliensis L. and C. laurifolius L.), considering the main stages after seed ingestion, i.e. seed recovery, seed germination, seedling emergence and early seedling establishment. Seed recovery after gut passage was high (around 40%) for all the species, except F. thymifolia (12%). Most seeds (ca. 90%) were recovered within 48 h after ingestion for all the species, although seeds were still recovered up to 96 h after ingestion. Gut passage increased germination up to seven-fold compared to non-ingested seeds. Furthermore, seedling emergence from seeds contained in pellets was overall similar (intact pellets) to or higher (crumbled pellets) than emergence from seeds without dung. Survival of emerged seedlings and mass of seedlings after 20 days were not reduced by dung. Sheep act therefore as effective dispersers of these Cistaceae species by scattering seeds and promoting germination, while faeces do not hamper seedling establishment. We conclude that the interaction between herbivorous ungulates and these dry-fruited species may be considered a mutualism qualitatively similar to the mutualism between frugivorous vertebrates and fleshy-fruited plants.     

GIGANTEA regulates phytochrome A-mediated photomorphogenesis independently of its role in the circadian clock

GIGANTEA (GI) is a nuclear protein involved in the promotion of flowering by long days, in light input to the circadian clock, and in seedling photomorphogenesis under continuous red light but not far-red light (FR). Here, we report that in Arabidopsis (Arabidopsis thaliana) different alleles of gi have defects in the hypocotyl-growth and cotyledon-unfolding responses to hourly pulses of FR, a treatment perceived by phytochrome A (phyA). This phenotype is rescued by overexpression of GI. The very-low-fluence response of seed germination was also reduced in gi. Since the circadian clock modulates many light responses, we investigated whether these gi phenotypes were due to alterations in the circadian system or light signaling per se. In experiments where FR pulses were given to dark-incubated seeds or seedlings at different times of the day, gi showed reduced seed germination, cotyledon unfolding, and activity of a luciferase reporter fused to the promoter of a chlorophyll a/b-binding protein gene; however, rhythmic sensitivity was normal in these plants. We conclude that while GI does not affect the high-irradiance responses of phyA, it does affect phyA-mediated very-low-fluence responses via mechanisms that do not obviously involve its circadian functions.     

The effects of seed size, cotyledon reserves, and herbivory on seedling survival and growth in Quercus rugosa and Q. laurina (Fagaceae)

In a greenhouse experiment, seedling survival of two oak species (Quercus rugosa and Q. laurina) was greatly affected by the excision of cotyledons 1 mo after germination, with a greater impact on Q. laurina. The effect of seed size was also significant for both species, with a positive correlation between seed mass and survival and growth. The effect of cotyledon excision on seedling growth persisted throughout the first growing season in Q. rugosa and was not analyzed for Q. laurina due to the low number of seedlings that survived cotyledon excision. Seed size significantly affected seedling height, diameter, leaf area, and biomass at 6 mo. Seed size and cotyledon retention affected the ability of Q. rugosa to recover from herbivory, as both factors had a significant effect on relative growth rates after aerial biomass removal. The results show that seedlings originating from large seeds can better endure loss of cotyledons and aerial biomass and thus are better equipped to confront stress early in their lives.     

外源水杨酸对盐胁迫下大黄种子萌发及幼苗生长的影响

该研究以掌叶大黄、唐古特大黄和药用大黄种子为材料,采用双层滤纸培养法,设置系列浓度NaCl (0、100、150、200、250 mmol/L) 胁迫试验,以及系列浓度水杨酸(SA)溶液(0、50、100、150、200、250 mg/L)拌种和浸种后盐胁迫实验,测定3种大黄种子萌发及幼苗生长指标,揭示外源水杨酸对盐胁迫下大黄种子萌发及幼苗生长的影响。结果显示：(1)随NaCl浓度增大3种大黄种子的发芽率均呈直线下降趋势,且子叶、胚轴、根和苗等生长均受到强烈抑制。(2)在拌种条件下, 200 mmol/L NaCl胁迫下掌叶大黄苗长在200 mg/L SA处理下受到显著促进; 200 mmol/L NaCl浓度盐胁迫下唐古特大黄种子发芽率在250 mg/L SA处理下受到显著抑制;100 mmol/L NaCl胁迫下药用大黄种子发芽势在200 mg/L SA处理下受到显著抑制,其发芽率在150 mg/L SA处理下得到显著抑制,其苗长在250 mg/L SA处理下受到显著抑制。(3)在浸种条件下, 200 mmol/L NaCl胁迫下掌叶大黄种子发芽率在50 mg/L SA处理下显著提高,其幼苗根长和苗长的生长在250 mg/L SA处理受到显著促进;200 mmol/L NaCl胁迫下唐古特大黄种子的发芽势在200 mg/L SA处理下得到显著促进,其幼苗根和苗的生长在50 mg/L SA处理下得到显著促进;100 mmol/L NaCl 胁迫下药用大黄根和苗的生长在100 mg/L SA处理下均得到显著促进。研究表明,3种大黄种子和幼苗对盐胁迫的响应趋势一致,但对不同浓度SA拌种和浸种的响应有较大差异。     

黄瓜种子及其萌发期的化感作用研究

以大白菜、萝卜、番茄和黄瓜种子为受体,采用实验室培养皿种子发芽生物测试法研究了黄瓜种子浸提液、种子萌发、胚根和芽苗分泌物、芽苗腐解物和芽苗浸提液的化感效应。结果表明:(1)黄瓜种子浸提液对大白菜、萝卜、番茄和黄瓜种子萌发均有化感抑制作用,即黄瓜种子内含有某些化感抑制物质。(2)在水浸提过的黄瓜种子萌发过程中,它不仅对其近邻套种的大白菜、萝卜和番茄种子萌发产生化感抑制作用,而且其胚根和芽苗分泌物对后茬播种的4种蔬菜种子发芽也表现出不同程度的化感抑制作用;黄瓜芽苗腐解物和芽苗水浸提液也对各受体蔬菜种子发芽与生长产生不同程度的化感抑制作用,且随着腐解芽苗量的增加或浸提液浓度的升高,各受体蔬菜种子的发芽指标值、化感效应指数值和综合效应值随之降低。(3)黄瓜种子浸提液及芽苗各器官的化感物质对黄瓜种子的萌发与生长产生了自毒作用,且黄瓜芽苗腐解物、芽苗浸提液、胚根及芽苗分泌物对受体黄瓜的自毒作用均为最大。研究发现,黄瓜种子浸提液、种子萌发时期以及芽苗各器官的化感物质主要是通过抑制受体胚根的生长而起化感抑制作用,即受体蔬菜种子胚根对化感效应最为敏感;因黄瓜种子及萌发期释放化感物质的途径有所不同,导致受体大白菜、萝卜、黄瓜和番茄的化感响应也不相同;在黄瓜种子萌发和芽苗生长的早期,化感物质即开始在芽苗体内进行合成与积累,一部分可通过胚根和芽苗分泌途径释放到环境中,另一部分可通过芽苗腐解途径释放化感物质,并对受体蔬菜种子萌发与生长表现出较强的化感抑制作用。     

Restionaceae seedlings: Morphology,anatomy and systematic implications

Restionaceae differ from most monocot families in having both epigeal and hypogeal germination. The green cotyledons associated with epigeal germination have a central vascular strand as found in most epigeal monocotyledons. In some genera the cotyledon may have a hairpin‐like structure, also described for Anthericaceae. The cotyledon of the hypogeal seedlings is short, without green pigment and largely remains embedded in the seed coat. Hypogeal germination is correlated with large, woody, indehiscent, frequently myrmecochorous nuts, while epigeal germination is found in species with smaller indehiscent nutlets or seeds, dispersed in a variety of ways. The primitive condition is most likely epigeal germination. In hypogeal seedlings of some African and Australian taxa an epicotyledonary rhizome is found between the primary root and the first leaves. Seedlings of African Restionaceae frequently have elongated culm internodes, whereas in the Australian species studied, internodes are very short, resulting in a cluster of seedling leaves. The leaf blades, which in most species are only found on the seedlings, are very simple anatomically. However, they appear to be unifacial, similar to the leaf blades of Anarthria (Anarthriaceae). The anatomical specialisations in the blades mirror those recorded for the culm anatomy. These observations are consistent with the hypothesis that Centrolepidaceae may be neotonous Restionaceae. They also corroborate the morphology of the African Restionaceae, and the presently accepted phylogeny of the African genera of Restionaceae.     

Seed reserve translocation and early seedling growth of eight tree species in a tropical deciduous forest in Mexico

Seed reserves play an essential role during germination and seedling establishment and are particularly important for species that grow in seasonal ecosystems with a short growing season. In this study, we examined (a) how and when the seedlings change their dependence from seed resources to external resources, (b) the lipid, nitrogen, and non-structural carbohydrate reserve translocation from seeds to seedlings over time, and (c) whether reserve translocation may be correlated to cotyledon and leaf lifespan of seedlings for eight tree species in a tropical deciduous forest in north-western Mexico. Our results showed that the cotyledon lifespan was not related to the cotyledon type (photosynthetic or reserve) and that the cotyledon biomass did not decrease significantly until germination. In six of the eight studied species, biomass allocation to the leaves was favored; lipids were the first reserve exhausted before the first leaves were totally expanded in seven of the eight study species. Species with the highest N concentration had expanded leaves and lost their cotyledons faster than species with a low N concentration. Our results suggest that tropical deciduous forest species employ different strategies to survive the dry season and re-sprout in the next growing season mediated by seed reserve concentrations, translocation patterns and subsequent biomass allocation.     

Seed size as an indicator of seed quality: a case study of Primula veris

Seed size is a widely accepted measure of seed quality, because many earlier studies have shown that large seeds have high seedling survival, growth and establishment. We tested whether ovule loss increases size of the remaining seeds and whether such size increase affects seedling establishment. We removed all except one flower from inflorescences of Primula veris L. (Primulaceae), a perennial hemicryptophyte herb, at a late stage of flowering. Flower removal (FR) increased seed size by 33% compared to the control plants. We then divided the seeds within each treatment to small, middle-sized and large seeds and carried out a sowing experiment in the field. Within each experimental group, seedling establishment was positively associated with seed size. However, despite size differences, seeds from the FR and control groups had the same seedling establishment probability. Seeds from FR plants had a higher seedling emergence in May than those from control plants, but the number of seedlings alive per sowing plot in the late summer was the same in both experimental groups. Increase in seed mass after partial FR thus did not enhance seedling performance, although seed size variation due to other causes was positively correlated with seedling establishment. Further studies are needed to show whether plastic changes of seed size are usually adaptive or not.     

Seed size as an indicator of seed quality: a case study of Primula veris

Seed size is a widely accepted measure of seed quality, because many earlier studies have shown that large seeds have high seedling survival, growth and establishment. We tested whether ovule loss increases size of the remaining seeds and whether such size increase affects seedling establishment. We removed all except one flower from inflorescences of Primula veris L. (Primulaceae), a perennial hemicryptophyte herb, at a late stage of flowering. Flower removal (FR) increased seed size by 33% compared to the control plants. We then divided the seeds within each treatment to small, middle-sized and large seeds and carried out a sowing experiment in the field. Within each experimental group, seedling establishment was positively associated with seed size. However, despite size differences, seeds from the FR and control groups had the same seedling establishment probability. Seeds from FR plants had a higher seedling emergence in May than those from control plants, but the number of seedlings alive per sowing plot in the late summer was the same in both experimental groups. Increase in seed mass after partial FR thus did not enhance seedling performance, although seed size variation due to other causes was positively correlated with seedling establishment. Further studies are needed to show whether plastic changes of seed size are usually adaptive or not.     

Seedling tolerance to cotyledon removal varies with seed size: A case of five legume species

It is generally accepted that seedlings from large seeds are more tolerant to defoliation than those from small seeds due to the additional metabolic reserves present in the large seeds. However, information on the effects of amount of seed reserves (cotyledon removal) from seedlings resulting from large vs. small seeds on seedling growth and long‐term survival in the field is limited. Five legume species with different sizes of seeds were sown in the field and none, one, or both cotyledons removed 7 days after seedling emergence. Seedling biomass, relative growth rate (RGR) and survival were determined at different time. Cotyledon removal, species, and their interaction had significant effects on seedling growth and survival. During the period between 33 and 70 days, seedlings from large seeds had a significantly lower RGR than those from small seeds. Biomass, RGR, and survival of seedlings from large seeds were significantly reduced by removal one or both cotyledons, whereas those of seedlings from small seeds were not affected. Seed energy reserves are more important for the early growth of seedlings from large seeds than for those from small seeds. The overall effect of cotyledon removal on growth and survival varies with seed size (i.e., energy reserves) with seedlings from small seeds being less sensitive than those from large seeds under field conditions.     

Seed size, seedling morphology, and response to deep shade and damage in neotropical rain forest trees

To investigate the existence of coordinated sets of seedling traits adapted to contrasting establishment conditions, we examined evolutionary convergence in seedling traits for 299 French Guianan woody plant species and the stress response in a shadehouse of species representing seed size gradients within five major cotyledon morphology types. The French Guianan woody plant community has larger seeds than other tropical forest communities and the largest proportion of hypogeal cotyledon type (59.2%) reported for tropical forests. Yet the community includes many species with intermediate size seeds that produce seedlings with different cotyledonal morphologies. A split-plot factorial design with two light levels (0.8% and 16.1% PAR) and four damage treatments (control, seed damage, leaf damage, stem damage) was used in the shadehouse experiment. Although larger-seeded species had higher survival and slower growth, these patterns were better explained by cotyledon type than by seed mass. Even larger-seeded species with foliar cotyledons grew faster than species with reserve-type cotyledons, and survival after stem grazing was five times higher in seedlings with hypogeal cotyledons than with epigeal cotyledons. Thus, to predict seedling performance using seed size, seedling morphology must also be considered.     

杉木种子与幼苗不同发育期的耐旱性研究

以广东杉木(Cunninghamia lanceolata)第二代改良种子园种子为试材,利用不同浓度聚乙二醇(PEG-6000)溶液(5%、10%、20%和40%)模拟不同程度(轻度、中度、较强、重度)干旱胁迫,并对不同干旱条件下的种子萌发、幼苗子叶期和真叶期不同发育阶段的形态变化进行测定分析。结果发现,杉木种子萌发初期对水分尤为敏感,耐旱性不强,尤其是随干旱程度的加剧,种子起始萌发时间不断后延且发芽率显著降低。子叶后期的杉木幼苗较子叶前期耐旱,真叶期的杉木幼苗对轻度和中度干旱胁迫具有较强的耐受性。     

Diaspore structure and germination ecophysiology of the babassu palm (Attalea vitrivir)

Both germination and seedling establishment in palm trees are strongly influenced by the morphoanatomy of the fruits, although the interactions of these processes with ecophysiological aspects are not yet well understood. The present work evaluated structural and physiological aspects of seed germination in Attalea vitrivir, a species living under the seasonal climate of the Cerrado (Brazilian savanna) biome. We studied morphology, anatomy and histochemistry of the fruits and seedlings, the effects on germination of the pericarp, of diaspore storage conditions, germination temperature, removal of the operculum, and of gibberellic acid (GA3) application, and characterized the imbibition process of the seeds. Germination depends on a series of complex interactions between structures and physiological processes. The pericarp protects the seed and also causes physical dormancy that, when interacting with the temperature regime, can delay germination until subsequent rainy periods. Some seeds demonstrated non-profound physiological dormancy associated with restricted germination imposed by the operculum. Germination and initial development result from the elongation of the cotyledon cells and are affected by the activities of two distinct meristems in the proximal region of the embryo. The haustorium develops an invaginated secretory epithelium and aerenchyma, and actively participates in seed reserve mobilization. The adaptation of A. vitrivir to the Cerrado environment, with a strongly seasonal climate, is favored by its diaspore structure, by the abundance of endosperm reserves that allow the seedlings to survive for a long period above soil level, and by a seedling development pattern that protects the vegetative axis by deep burial.     

干旱胁迫下华北驼绒藜种子大小及苞片对萌发和幼苗生长的影响

干旱是影响荒漠区植物种子萌发和幼苗生长的关键因素。以多年生强旱生半灌木华北驼绒藜为对象,研究了不同干旱程度(0、100、200、300和400 g·L^-1 PEG6000)下,种子大小及苞片有无对种子萌发及幼苗生长的影响。结果表明: 干旱胁迫显著抑制了种子萌发和幼苗地上部的生长。100和200 g·L^-1 PEG6000处理显著增加了幼苗根长,而300和400 g·L^-1 PEG6000处理显著降低了幼苗根长。与无苞片相比,有苞片使种子的发芽率显著降低12%,发芽指数显著降低50.5%,幼苗的地上部长度显著增加20.8%,幼苗根长显著增加6.3%。种子大小对种子发芽指数无显著影响,但与小种子相比,大种子发芽率显著提高3%,幼苗的地上部长度显著增加20.5%,幼苗根长显著增加33.0%。在干旱条件下,苞片能延缓种子的萌发速度,种子大小能影响后代的存活几率,二者共同影响华北驼绒藜对极端干旱环境的适应性。     

Seed germination patterns in green dragon (Arisaema dracontium,Araceae)

Arisaema dracontium (green dragon) is a perennial herb that is widely distributed in eastern North America. However, in Canada, at the northern edge of its distribution, the species is designated as “vulnerable” with respect to conservation status. In natural populations, seedlings are uncommon; the present study was undertaken in order to characterize seed and seedling properties in green dragon. Seeds were sampled from five sites, ranging from Ontario at the northern limit of the distribution range, to Louisiana in the south. Seed germinability ranged from 25 to 55%, depending upon source. Experiments indicated that neither the hard seed coat nor a water-soluble exudate from the seed was responsible for inducing or maintaining dormancy. Patterns of seed germination appear to reflect general climatic conditions at the sites where seeds had originated. Cold stratification at 3°C produced significantly greater relative germinability in all seed collections except the most southerly one, from Baton Rouge. These seeds also had a slower overall speed of germination. In contrast, germination of seeds from the most northerly site was promoted by cold stratification and occurred over a relatively brief period. Germination in alternating light and dark conditions decreased the speed of germination compared to germination in the dark, however exposure to light changed the phenology of germination by promoting development of adventitious roots and primary leaves in these seedlings.