The performance of carrot seeds in relation to their viability

The germination and emergence characteristics, and early seedling growth, of carrot seeds cv. Chantenay red-cored from different sources with a range of germination from 54–94%, was compared. Seeds from protected crops (mean temperature of growth 21°C, r.h. 45–70%) gave higher percentage germination than those from crops grown outdoors (mean temperature 15°C, 70–100% r.h.). Germination was also higher from mature (seed moisture content at harvest 20% or lower) than immature seed (seed moisture content at harvest between 20 and 60%). High percentage germination (>90%) was associated with low mean germination times and low spreads of germination times whilst the reverse was true for low percentage germination. Similar relationships were found for seedling emergence characters in the field although a lower proportion of the viable seeds produced seedlings from slowly than rapidly germinating seed lots. In general, seed lots having a low percentage germination gave greater variability in plant weight than those of higher percentage germination. There was no effect of seed source on radicle or shoot relative growth rates or on post-emergence seedling growth rates.     

A study of the relationship between seedling characters and rate of germination within a seed lot

The relationship between seedling characters and germination rate within a seed lot was studied in cauliflower, leek and onion seeds. Newly germinated seeds were selected after successive days of imbibition at 20°C and placed on slope tests to assess early seedling growth. In all three species seedling length decreased and the coefficient of variation of those seedling lengths increased with increasing number of days of imbibition required for germination. Slow germinating seeds in all three species produced fewer normal healthy seedlings than faster germinating seeds. The relevance of these results to pre-germinated seed sowing techniques is discussed.     

Patterns of protein oxidation in Arabidopsis seeds and during germination

Increased cellular levels of reactive oxygen species are known to occur during seed development and germination, but the consequences in terms of protein degradation are poorly characterized. In this work, protein carbonylation, which is an irreversible oxidation process leading to a loss of function of the modified proteins, has been analyzed by a proteomic approach during the first stages of Arabidopsis (Arabidopsis thaliana) seed germination. In the dry mature seeds, the legumin-type globulins (12S cruciferins) were the major targets. However, the acidic alpha-cruciferin subunits were carbonylated to a much higher extent than the basic (beta) ones, consistent with a model in which the beta-subunits are buried within the cruciferin molecules and the alpha-subunits are more exposed to the outside. During imbibition, various carbonylated proteins accumulated. This oxidation damage was not evenly distributed among seed proteins and targeted specific proteins as glycolytic enzymes, mitochondrial ATP synthase, chloroplastic ribulose bisphosphate carboxylase large chain, aldose reductase, methionine synthase, translation factors, and several molecular chaperones. Although accumulation of carbonylated proteins is usually considered in the context of aging in a variety of model systems, this was clearly not the case for the Arabidopsis seeds since they germinated at a high rate and yielded vigorous plantlets. The results indicate that the observed specific changes in protein carbonylation patterns are probably required for counteracting and/or utilizing the production of reactive oxygen species caused by recovery of metabolic activity in the germinating seeds.     

Patterns of protein synthesis during the germination of pea axes,and the effects of an interrupting desiccation period

As germination of axes of Pisum sativum L. seeds progressed, profound quantitative and qualitative changes occurred in the patterns of protein synthesis. This was shown by fluorography of gels following two-dimensional polyacrylamide gel electrophoresis separation of [³⁵S]methioninelabelled proteins. The effects of desiccation during germination on these in-vivo protein-synthesis patterns were followed. Desiccation differentially affected the synthesis of proteins. Usually, however, upon rehydration following desiccation the types of proteins being synthesized were recognizable as those synthesized earlier during imbibition of control, once-imbibed axes: seeds imbibed for 8 h, and then dried, did not recommence synthesis of proteins typical of 8-h-imbibed control seeds, but rather of 4-h-imbibed control seeds. Seeds imbibed for 12 h, and then dried and rehydrated, synthesized proteins typical of 4-h-and 8-h-control seeds. Thus drying of germinating pea axes caused the proteinsynthesizing mechanism to revert to producing proteins typical of earlier stages of imbibition. Drying during germination never caused the seed to revert to the metabolic status of the initial mature dry state, however.Abbreviation DR dried and rehydrated     

Phytotoxic effects of fumonisin B1 on maize seedling growth

Fumonisin B₁ toxin is produced by the fungusFusarium moniliforme Sheldon, which is systemic to maize (Zea mays L.) and maize seeds. The effects of zero to 100 parts per million fumonisin B₁ on the germination process of maize seeds was determined. The presence of fumonisin had no effect on percent seed germination, but fumonisin inhibited radicle elongation by up to 75% after 48 hours of imbibition. An analysis of amylase secretion in the maize endosperm indicated that fumonisins inhibited amylase production in the germinating seed. Isoelectric focusing of endosperm extracts indicated that secretion of the low pI class of amylases was affected more that other amylase isozymes. The results suggested that the presence of high levels of fumonisin in maize seed may have deleterious effects on seedling emergence.     

Imbibition of white spruce seeds and somatic embryos: A study of morphological changes in an environmental scanning electron microscope and potassium leakage

Summary Potassium leakage and morphological changes during imbibition of white spruce [Picea glauca (Moench) Voss] seeds and somatic embryos were investigated. A single desiccated somatic embryo, a single somatic embryo exposed to a high relative humidity environment for 2 d, and a single dry zygotic embryo leaked similar amounts of potassium over a 120-min period of imbibition in liquid germination medium. A seed without a seed coat leaked two and eight times more potassium than a single whole seed and a single zygotic embryo, respectively. Nearly 50% of the potassium leaked for all tissues was leaked within the first 20 min of imbibition. Exposure of somatic embryos to an environment with high relative humidity resulted in a reduction in the percentage of potassium leaked after 80 and min to levels equivalent to those for zygotic embryos. Using an environmental scanning electron microscope, we found that desiccated somatic embryos and dry zygotic embryos had wrinkled surface cells, with cells in the surface of zygotic embryos being more shrunken in appearance. Imbibition of both types of embryos in water resulted in turgid surface cells after 2 h. Imbibition in liquid germination medium did not cause much hydration of surface cells, which still had wrinkled appearances after 2 h. Finally, imbibition on filter paper on semisolidified germination medium resulted in slower hydration of somatic and zygotic embryos. Cells near the medium appeared hydrated while cotyledon surface cells furthest from the medium resembled cells in desiccated embryos.     

Investigation of the relationship between seed leachate conductivity and the germination of Brassica seed

Large differences in the leachate conductivities of single seeds after 24 h soaking were observed in five Brussels sprout cultivars. Within each cultivar non-germinable seeds tended to have higher conductivities than germinable seeds, but the conductivity above which seeds failed to germinate varied between cultivars. No conductivity reading or partition value could therefore be considered as an absolute boundary between germinable and non-germinable seeds to allow the prediction of seed germination for all seed lots of the same species. However, the mean conductivity for each sample was significantly correlated with germination (r = 0.83, P < 0.05) and was clearly higher for samples with lower germinations. Differences in the partition values for the five cultivars may have been attributable to the natural age of the seeds. Seeds of Asmer Aries artificially aged rapidly at 20% moisture content for up to 48 h at 45° C showed increasing leachate conductivity with increased time of ageing (r = 0.97, P < 0.01) and decreased germination (r = - 0.88, P < 0.05). Again however the values for germinable and non-germinable seeds of different artificial ages varied. Leakage from seeds was limited by the testa in the first 6 h of imbibition. Solute leakage from embryos of the five cultivars was therefore greater than those for intact seeds but a clear separation of germinable and non-germinable seeds was still not possible from the conductivity readings. The implications of the observed increases in conductivity with increased seed age and decreased germination for the development of a rapid vigour test for small seeded vegetables in conjunction with controlled deterioration are discussed.     

Ascorbate and glutathione metabolism in embryo axes and cotyledons of germinating lupine seeds

Changes in ascorbate and glutathione contents and the activities and isoenzyme patterns of enzymes of the ascorbate-glutathione cycle were investigated in embryo axes and cotyledons of germinating lupine (Lupinus luteus L.) seeds. Ascorbate content was not significantly affected over the initial 12 h of imbibition in embryo axes, but afterwards increased, with the most rapid accumulation coinciding with radicle emergence. A somewhat similar trend was observed for glutathione with significant increase in embryo axes shortly before radicle protrusion followed by decline in the next hours. In cotyledons the ascorbate pool rose gradually during germination but the amount of glutathione showed fluctuations during a whole germination period. The activity of ascorbate peroxidase (APX) rose progressively in embryo axes, while activities of dehydroascorbate reductase (DHAR) and glutathione reductase (GR) showed transient increase during germination. New isoforms of APX and GR were synthesized, suggesting that they play a relevant role during germination. All analyzed enzymes were already present in dry seeds which allowed them to be active immediately after imbibition.     

Pepper seed germination assessed by combined X-radiography and computer-aided imaging analysis

A lot of pepper seeds having 87 % germination were subjected to X-ray inspection using a non lethal dose of radiation. Seeds with less than 2.7 % (on the basis of total seed area) of free space area, i.e. the spaces between embryo and endosperm, were classified as highly viable seeds (97–100 % germination) with the lowest level of abnormal seedlings. Seeds X-ray classified as good were subjected to a computerised image analysis to study seed imbibition and radicle elongation. The patterns of seed area increase, chosen as the most accurate indicator of seed swelling, resembled the triphasic curve of water uptake. The first phase was completed at 9 h followed by a second phase that varied widely in time until completion of germination between 52 and 96 h. The proportion of seeds with radicle protrusion between 52–56 h and 64–72 h assessed with the image analysis was significantly higher than that recorded using a conventional germination test. In addition, the rate of increase of seed area during the third phase of imbibition, mostly due to protrusion of the radicle tip and its growth, was highly correlated with the corresponding radicle elongation rate.     

Relationship between the ATP content measured at three imbibition times and germination of onion seeds during storage at 3, 15 and 30C

The possibility of using ATP content as an indicator of seedquality was studied in onion seeds {Allium cepa cv. Wdenswil).The percentage germination and ATP content of imbibed seedswere compared during 145 weeks of storage at three temperatures(3, 15 and 30C). ATP content, which was undetectable in airdriedseeds (moisture content: 9%, w/w), increased rapidly as a functionof imbibition time, as did the fresh weight and respirationrate, reaching a steady-state level after about 17 h. After36 weeks of storage, the rate of ATP formation was greater forthe seeds stored at 3C than for those kept at 15 and 30C.Furthermore, the onset of ATP synthesis was delayed. These phenomena,which are likely to be an expression of seed ageing, are usefulindicators, allowing the prediction of the loss of seed viabilitybefore the decrease in percentage germination which occurredbeyond 36 weeks of storage. In addition, the correlation betweenATP content and germination capacity of seeds during 145 weeksof storage was excellent (r=0.95 at 15C and 0.97 at 30C),provided that a 17 h-imbibition time, specific for onion seeds,was chosen. These results are discussed in terms of the controversyconcerning the correlation between the ATP content and germinationpercentage of seeds. Key words: Allium cepa, ageing, bioenergetic metabolism, seed quality, temperature storage     

Changes of Adenylate Pool and Energy Charge E. C. in Relation to the Germination of Chinese Cabbage Seeds

The changes of adenylate pool (ATP, ADP, AMP) and energy charge (E. C.) were determined in germinating seeds of Chinese cabbage. The dry seeds contained a few ATP, most of the adenylate was in the form of AMP. After 1 h of inhibition, most of AMP was converted to ATP, and E. C. value increased rapidly. If the inhibition continued for two hours, the level of ATP increased about 35-fold, and E. C. value reached 0.51. After 24 h of germination, the ATP level in the non- germinating seeds was lower 4 times than that in the germinating seeds and E. C. value was lower too. If the seeds germinated in 3% O2, the ATP level dropped. As the seeds were transfered to air, the adenylate pool and E. C. value increased to the level of the control, then the percentage germination reached 76%. When the seeds were treated with 3% O2, 5 × 10-4 M 2, 4-dinitro phenol, 1 × 10-5 M earbonyl cyanid m-chlorophenyl hydrozone and 5 × 10-3 M iodoaeetie acid at the first 1–2 h, they inhibited the production of ATP and decreased E. C. value. Iodoaeetic acid was the most effective one among the four inhibitors. It was shown that the rate of germination was closely related to the energy charge and the adenylate pool.     

Differential tissue-specific expression of cysteine proteinases forms the basis for the fine-tuned mobilization of storage globulin during and after germination in legume seeds

The temporal and spatial distribution of cysteine proteinases (CPRs) was analyzed immunologically and by in situ hybridization to identify the CPRs involved in the initiation of storage-globulin degradation in embryonic axes and cotyledons of germinating vetch (Vicia sativa L.). At the start of germination several CPRs were found in protein bodies in which they might have been stored in the mature seeds. Cysteine proteinase 1 was predominantly found in organs like the radicle, which first start to grow during germination. Cysteine proteinase 2 was also present at the start of germination but displayed a less-specific histological pattern. Proteinase B was involved in the globulin degradation of vetch cotyledons as well. The histological pattern of CPRs followed the distribution of their corresponding mRNAs. The latter were usually detected earlier than the CPRs but the in situ hybridization signals were histologically not as restricted as the immunosignals. Proteolytic activity started in the radicle of the embryonic axis early during germination. Within 24 h after imbibition it had also spread throughout the whole shoot. At the end of germination, newly synthesized CPRs might have supplemented the early detectable CPRs in the axis. In the cotyledons, only the abaxial epidermis and the procambial strands showed proteinase localization during germination. Both CPR1 and CPR2, as well as the less common proteinase B, might have been present as stored proteinases. Three days after imbibition, proteolytic activity had proceeded from the cotyledonary epidermis towards the vascular strands deeper inside the cotyledons. The histochemical detection of the CPRs was in accordance with the previously described histological pattern of globulin mobilization in germinating vetch [Tiedemann J, et al. (2000)]. A similar link between the distribution of CPRs and globulin degradation was found in germinating seeds of Phaseolus vulgaris L. The coincidence of the histological patterns of globulin breakdown with that of the CPRs indicates that at least CPR1, CPR2 and proteinase B are responsible for bulk globulin mobilization in the seeds of the two legumes. Received: 14 February 2000 / Accepted: 16 August 2000     

Seed vigour and water relations in wheat

The laboratory germination (criterion radicle emergence) of seven seed lots of winter wheat cv. Slejpner was similar. However, they differed in vigour as demonstrated by differences in germination after controlled deterioration carried out at a range of seed moisture contents, at two temperatures and for different times. A vigour assessment for each lot was quantified by calculating a value for the seed lot constant K_i, of the viability equation. Germination in lower water potentials reduced the uptake of water, radicle and coleoptile emergence and radicle and coleoptile extension. There was no difference in the water uptake of seed lots differing in vigour. However, seed lots of lower vigour showed less radicle emergence, less coleoptile emergence and shorter radicles than higher vigour seed lots in low water potentials. Similarly, controlled deterioration resulted in reduced radicle and coleoptile emergence and growth compared to unaged seed, and also to a greater sensitivity to low water potentials. The implications for field establishment are discussed.     

Inter-nucleosomal DNA fragmentation and loss of RNA integrity during seed ageing

The germination of viable seeds is the basis for new plant growth and development. Seeds lose viability during storage, but the biochemical mechanisms of seed death are not fully understood. This study aimed to investigate degradation patterns of nucleic acids during seed ageing and subsequent water uptake. Seeds of Pisum sativum L. were artificially aged at 50°C and 12% seed water content (WC). Nucleic acids degradation was studied during ageing and during imbibition of four seed lots with differential viability from highly viable to dead. As seeds lost viability during ageing, DNA was gradually degraded into internucleosomal fragments, resulting in ‘DNA laddering’, in conjunction with disintegration of 18S and 28S rRNA bands. During imbibition, non-aged controls had high levels of DNA and RNA integrity through to radicle protrusion. In an aged seed lot with 85% total germination (TG) DNA fragmentation decreased upon imbibition probably due to nucleosome degradation, while rRNA integrity did not improve. In an aged seed lot with 44% TG, neither DNA nor rRNA integrity improved upon imbibition. Dead seeds showed DNA degradation as laddering throughout imbibition along with extensive degradation of rRNA. We present a model in which interlinked programmed and non-programmed events contribute to seed ageing, and suggest that protection of nucleic acids during ageing is key to seed longevity.     

Spatio-temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer, and endosperm tissues

Germination of barley is accompanied by changes in water-soluble seed proteins. 2-DE was used to describe spatio-temporal proteome differences in dissected seed tissues associated with germination and the subsequent radicle elongation. Protein identification by MS enabled assignment of proteins and functions to the seed embryo, aleurone, and endosperm. Abundance in 2-DE patterns was monitored for 48 different proteins appearing in 79 gel spots at 8 time-points up to 72 h post imbibition (PI). In embryo, a beta-type proteasome subunit and a heat shock protein 70 fragment were among the earliest proteins to appear (at 4 h PI). Other early changes were observed that affected spots containing desiccation stress-associated late embryogenesis abundant and abscisic acid (ABA)-induced proteins. From 12 h PI proteins characteristic for desiccation stress disappeared rapidly, as did a putative embryonic protein and an ABA-induced protein, suggesting that these proteins are also involved in desiccation stress. Several redox-related proteins differed in spatio-temporal patterns at the end of germination and onset of radicle elongation. Notably, ascorbate peroxidase that was observed only in the embryo, increased in abundance at 36 h PI. The surprisingly early changes seen in the protein profiles already 4 h after imbibition indicate that germination is programmed during seed maturation.     

A gibberellin-regulated xyloglucan endotransglycosylase gene is expressed in the endosperm cap during tomato seed germination.

Xyloglucan endotransglycosylases (XETs) modify xyloglucans, major components of primary cell walls in dicots. A cDNA encoding an XET (LeXET4) was isolated from a germinating tomato (Lycopersicon esculentum Mill.) seed cDNA library. DNA gel blot analysis showed that LeXET4 is a single-copy gene in the tomato genome. LeXET4 mRNA was strongly expressed in germinating seeds, was much less abundant in stems, and was not detected in roots, leaves or flower tissues. During germination, LeXET4 mRNA was detected in seeds within 12 h of imbibition with maximum mRNA abundance at 24 h. Tissue prints showed that LeXET4 mRNA was localized exclusively to the endosperm cap region. Expression of LeXET4 was dependent on exogenous gibberellin (GA) in GA-deficient (gib-1 mutant) tomato seeds, while abscisic acid, a seed germination inhibitor, had no effect on LeXET4 mRNA expression in wild-type seeds. LeXET4 mRNA disappeared after radicle emergence, even though degradation of the lateral endosperm cell walls continued. The temporal, spatial and hormonal regulation pattern of LeXET4 gene expression suggests that XET has a role in endosperm cap weakening, a key process regulating tomato seed germination.     

Activities of Hydrogen Peroxide-Scavenging Enzymes in Germinating Wheat Seeds

During imbibition and germination of wheat (Triticum aestivum)in the dark over 72 h, activities of the enzymes of the ascorbate(AsA)-dependent H₂O₂-scavenging pathway, AsA peroxidase, monodehydroascorbate(MDAsA) reductase, dehydroascorbate (DHAsA) reductase and glutathione(GSSG) reductase as well as superoxide dismutase (SOD), catalaseand guaiacol peroxidase were determined both in whole grainsand in isolated embryos and endosperm. With the exception of DHAsA reductase, activities of the otherenzymes assayed increased in germinating seeds, especially duringradicle emergence (between 24–48 h of imbibition). Theseincreases, particularly for AsA peroxidase, were much higherin the embryo than in the endosperm. Within 72 h of imbibition,activities per seed increased 116-fold for AsA peroxidase, 19-foldfor guaiacol peroxidase, 5-fold for catalase and only 1·4-foldfor SOD. In contrast to the decreases in DHAsA reductase, theother AsA recycling enzyme, MDAsA reductase, increased 5-foldwithin 72 h. The results indicate that, in wheat seeds, imbibition and germinationis associated with enhanced cellular capacity to detoxify H₂O₂.For this detoxification the operation of AsA peroxidase togetherwith the AsA-regenerating enzymes appears to be of particularimportance. Key words: Ascorbate peroxidase, germination, hydrogen peroxide detoxification, inhibition, wheat     

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 ).