Seed Development in a Shrunken Endosperm Barley Mutant

Seg8 is one of eight barley (Hordeum vulgare L.) mutants whoseendosperm development is affected by the maternal plant genotype.This study was initiated to determine the nature and onset ofabnormal development to provide a basis for further studiesaimed at understanding the mechanism of genetic control. Seeddevelopment and synthesis and accumulation of reserve substanceswere compared between seg8 and its normal counterpart, cv. Klages.Light microscopic examination showed that the mutant phenotypeappeared as early as 4 d after anthesis (DAA), and seg8 graindry weight was significantly lower than cv. Klages by 8 DAA.Grain cell number was significantly lower in seg8 by 8 DAA,indicating an early termination of cell division. The mutanthad a lower starch concentration and higher sucrose concentration,also evident at 8 DAA. Rates of [¹⁴C]sucrose incorporation intostarch in excised half seeds were similar in both genotypesat 2 and 4 DAA, but at 8 and 12 DAA seg8 had a lower rate. Totalprotein concentration was not significantly different betweenthe two genotypes throughout endosperm development. These resultsindicate that the mutation affects cell division and starchaccumulation prior to 8 DAA. It is not known if the reductionin starch biosynthesis and accumulation results from a reducedcapacity for starch or a defect in starch biosynthesis. Hordeum vulgare L., barley, shrunken endosperm mutant, endosperm development, starch, protein, endosperm cell number     

小麦籽粒发育及淀粉晶体特性对花后高温胁迫的响应

选用2个品质类型和成熟期不同的新疆主栽小麦品种‘新春11号’和‘新春39号’,分别进行花后灌浆早期高温(花后5~8d,32℃,T_1)和中期高温(花后15~18d,38℃,T_2)处理,分析花后高温对小麦籽粒发育及淀粉晶体的影响。结果显示:(1)T_1处理明显降低了两品种籽粒长度和粒重,而T_2处理显著影响籽粒宽度和厚度;高温处理虽然降低了籽粒灌浆速率,但两品种灌浆最大峰值出现时间均在花后18d。(2)T_1处理对小麦籽粒A型淀粉粒形态的影响较大,中熟品种‘新春11号’的A型淀粉粒表面在花后10d时可观察到微孔,在花后15~20d时其粒径明显小于同期对照,在花后20~25d时淀粉粒表面压痕增多且A、B型淀粉粒表面出现明显缢缩;而早熟品种‘新春39号’淀粉粒形态和粒径大小受花后高温的影响相对较小。(3)两品种在不同高温处理下,其淀粉粒晶体特性衍射峰出现的位置相同,但淀粉粒的尖峰强度不同,表明高温胁迫不影响淀粉粒的晶体类型,但可能改变了淀粉粒内部的层状结构。研究表明,花后早期高温不仅对小麦籽粒外部形态有较大的影响,同时也影响到籽粒内部淀粉粒的形态和晶体的特性。     

ADP-ribose gates the fertilization channel in ascidian oocytes

We report an ionchannel in the plasma membrane of unfertilized oocytes of the ascidianCiona intestinalis that is directly gated by the second messenger ADP-ribose. The ion channel is permeable to Ca²⁺ andNa⁺ and ischaracterized by a reversal potential between 0 and +20 mV and aunitary conductance of 140 pS. Preinjection of theCa²⁺ chelator1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) or antagonists of intracellularCa²⁺ release channels into oocytesdid not inhibit the ADP-ribose current, demonstrating that the channelis activated in a Ca²⁺-independentmanner. Both the fertilization current and the current induced by theinjection of nicotinamide nucleotides are blocked by nicotinamide,suggesting that the ADP-ribose channel is activated at fertilization ina nicotinamide-sensitive manner. These data suggest that ascidian spermtrigger the hydrolysis of nicotinamide nucleotides in the oocyte toADP-ribose and that this mechanism is responsible for the production ofthe fertilization current.

     

Differences of Starch Granule Distribution in Grains from Different Spikelet Positions in Winter Wheat

Wheat starch development is a complex process and is markedly difference by changes in spikelet spatial position. The present study deals with endosperm starch granule distribution and spatial position during filling development. The study was conducted with pure starch isolated from wheat (Triticum aestivum L.), Jimai20 and Shannong1391, at 7–35 days after anthesis (DAA). The results showed that grain number, spikelet weight and grain weight per spikelet in different spatial position showed parabolic changes. Upper spikelets had highest starch and amylose content followed by basal spikelets, then middle spikelets. The paper also suggested the volume percents of B-type and A-type granule in grain of middle spikelets were remarkably higher and lower than those of basal and upper spikelets, respectively. However, no significant difference occurred in the number percents of the two type granule. The ratio of amylase to amylopectin was positively correlated with the volume proportion of 22.8–42.8 µm, but was negatively related to the volume proportion of <9.9 µm. The results indicated that the formation and distribution of starch granules were affected significantly by spikelet position, and grains at upper and basal spikelet had the potential of increasing grain weight through increasing the volume of B-type granules.     

Spatial distribution of proteins and metabolites in developing wheat grain and their differential regulatory response during the grain filling process

Grain filling and grain development are essential biological processes in the plant’s life cycle, eventually contributing to the final seed yield and quality in all cereal crops. Studies of how the different wheat (Triticum aestivum L.) grain components contribute to the overall development of the seed are very scarce. We performed a proteomics and metabolomics analysis in four different developing components of the wheat grain (seed coat, embryo, endosperm, and cavity fluid) to characterize molecular processes during early and late grain development. In-gel shotgun proteomics analysis at 12, 15, 20, and 26 days after anthesis (DAA) revealed 15 484 identified and quantified proteins, out of which 410 differentially expressed proteins were identified in the seed coat, 815 in the embryo, 372 in the endosperm, and 492 in the cavity fluid. The abundance of selected protein candidates revealed spatially and temporally resolved protein functions associated with development and grain filling. Multiple wheat protein isoforms involved in starch synthesis such as sucrose synthases, starch phosphorylase, granule-bound and soluble starch synthase, pyruvate phosphate dikinase, 14-3-3 proteins as well as sugar precursors undergo a major tissue-dependent change in abundance during wheat grain development suggesting an intimate interplay of starch biosynthesis control. Different isoforms of the protein disulfide isomerase family as well as glutamine levels, both involved in the glutenin macropolymer pattern, showed distinct spatial and temporal abundance, revealing their specific role as indicators of wheat gluten quality. Proteins binned into the functional category of cell growth/division and protein synthesis/degradation were more abundant in the early stages (12 and 15 DAA). At the metabolome level all tissues and especially the cavity fluid showed highly distinct metabolite profiles. The tissue-specific data are integrated with biochemical networks to generate a comprehensive map of molecular processes during grain filling and developmental processes.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.): VI. INFLUENCE OF PRIMING ON GERMINATION RESPONSES TO TEMPERATURE AND WATER POTENTIAL DURING SEED DEVELOPMENT

Seed priming (imbibition in water or osmotic solutions followedby redrying) generally accelerates germination rates upon subsequentre-imbibition, but the response to priming treatments can varyboth within and among seed lots. Seed maturity could influenceresponsiveness to priming, perhaps explaining variable primingeffects among developmentally heterogeneous seed lots. In thecurrent study, muskmelon (Cucumis melo L.) seeds at two stagesof development, maturing (40 d after anthesis (DAA)) and fullymature (60 DAA), were primed in 0?3 M KNO₃ for 48 h at 30 ?C,dried, and imbibed in polyethylene glycol 8000 solutions of0 to –1?2 MPa at 15, 20, 25, and 30 ?C. Germination sensitivitiesto temperature and water potential () were quantified as indicatorsof the influence of seed maturity and priming on seed vigour.Germination percentages of 40 and 60 DAA control seeds weresimilar in water at 30 ?C, but the mean germination rate (inverseof time to germination) of 40 DAA seeds was 50% less than thatof 60 DAA seeds. Germination percentages and rates of both 40and 60 DAA seeds decreased at temperatures below 25 ?C. Reductionsin also delayed and inhibited germination, with the 40 DAAseeds being more sensitive to low than the 60 DAA seeds. Primingsignificantly improved the performance of 40 DAA seeds at lowtemperatures and reduced , but had less effect on 60 DAA seeds.Priming lowered both the minimum temperature (T_b) and the minimum (_b) at which germination occurred. Overall, priming of 40 DAAseeds improved their germination performance under stress conditionsto equal or exceed that of control 60 DAA seeds, while 60 DAAseeds exhibited only modest improvements due to priming. Asthe osmotic environment inside mature fruits approximates thatof a priming solution, muskmelon seeds may be ‘primed’in situ during the late stage of development after maximum dryweight accumulation. Key words: Cucumis melo L., seed priming, germination, vigour, development, temperature     

Activity of the Pentose Phosphate Pathway and Changes in Nicotinamide Nucleotide Content during Germination of Seeds of Cicer arietinum L.

Changes in the level of nicotinamide nucleotides, rate of ¹⁴CO₂output from [1^–14C] or [6¹⁴ C₆/C₁ ratios, glucose-6-phosphatedehydrogenase, 6-phosphogluconate dehydrogenase, and NAD kinaseactivities were determined during the first 72 h of germinationof seeds of Cicer arietinum L. The level of oxidized and reducedforms of nicotinamide nucleotides, together with the activityof glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase,NAD kinase, and C₆/C₁ ratios, suggest that the pentose phosphatepathway is activated during early germination in cotyledonsof chick pea seeds. The results obtained in embryonic axes seemsto indicate a lower participation of the PP pathway, probablydue to the development of the activity of the glycolytic-TCApathway.     

Effect of High Temperature During Grain-filling on the Structure of Developing and Malted Barley Grains

High temperatures (up to 35 °C) were applied to plants ofmalting barley,Hordeum vulgareL. (‘Schooner’) fora period of 5 d during grain-filling. Heat treatment had a profoundeffect on the structure of the mature barley grain. There wasevidence of degradation of endosperm storage products in heat-treatedgrain. Starch granule development was reduced in sub-aleuronecells following heat treatment and alterations to starch granuledistribution and growth were observed in the endosperms of thesegrains. Endosperm cell wall and crushed cell layer (CCL) developmentwere sensitive to high temperatures, with the reduced thicknessof the CCL and generally patchy Calcofluor fluorescence of endospermcell walls indicative of partial hydrolysis of ß-glucans.Increased growth of the embryo took place in heat-treated grainscompared with control grains. Endosperm texture was generallymore friable in heat-treated grains than in control grains,and these grains overmodified during malting, with considerabledegradation of starch in the form of extensive pitting of A-typestarch granules. Evidence is presented for developmental andgermination events occurring simultaneously within the developinggrain.Copyright 1998 Annals of Botany Company Barley,Hordeum vulgareL., starch granules, crushed cell layer, scutellum, embryo, fluorescence microscopy, scanning electron microscopy, confocal microscopy, malting quality.     

Effect of Post-anthesis Drought on Cell Division and Starch Accumulation in Developing Wheat Grains

Wheat plants (Triticum aestivum L., cv. Warigal) were subjectedto 20 d of water deficit during the period of endosperm celldivision. Drought accentuated the differences in final grainweight between spikelets and between grains within spikelets.The distal grains of top spikelets were most affected by drought.The maximum number of endosperm cells was, respectively, 30and 40 per cent lower in basal grains and distal grains of draughtedplants. In basal grains of middle spikelets, the number of largestarch granules per cell was unaffected but the number of smallstarch granules per cell was 45 per cent lower in grains ofdraughted plants. The initiation of small starch granules wasmore affected than cell division because severe water deficitoccurred earlier during the former process than the latter.Final dry weight appeared to correlate well with the maximumnumber of endosperm cells, but depended also on the number ofstarch granules per cell. Consequently, the amount of dry matterper cell was not constant in both treatments. The concentration of sucrose per endosperm cell was lower onlyin the droughted distal grains of top spikelets. The supplyof sucrose to endosperm cells did not regulate the initiationof small starch granules. Triticum aestivum L., wheat, drought, grain growth, cell division, starch     

Regulation of Grain Weight by Supply of Assimilates and Starch Granule Development in Three Winter Wheat Varieties

On removing the top half of the ear (halving) on several datesafter anthesis, dry weight per grain increased in three winterwheat (Triticum aestivum L.) varieties, in two pot experiments;the increase was greater with early than with late halving.The variety Splendeur had a lower dry weight and water percentagein grains than either Hobbit or Maris Huntsman. The ratio ofthe green area integrated over the post-anthesis period to thenumber of grains per ear (green area per grain) was highestin Splendeur and lowest in Hobbit in the first experiment; inthe second, Splendeur gave a lower ratio than the other twovarieties, which showed similar values. The green area per grainwas greater the earlier the ear was halved. The number of A-typestarch granules per endosperm, but not the volume per A granule(modal volume) and the modal volume of B starch granules, butnot their number, increased to a greater extent with early thanwith late halving. In Splendeur the grains had fewer A starchgranules, although these were of greater modal volume than inHobbit and Maris Huntsman and a number of B starch granulessimilar to the other two varieties, but of smaller modal volume.Maris Huntsman had more A granules than Hobbit, but with smallermodal volume. Dry weight per grain increased linearly with thenumber of A starch granules per endosperm, which in turn increasedasymptotically with green area per grain. The regressions forthe three varieties differed significantly. The influence ofthe supply of assimilates and the capacity for starch granuleformation in the regulation of grain weight is discussed. Key words: Grain weight, starch granules, assimilate supply, variety, wheat     

Antifungal Proteins during Sorghum Grain Development and Grain Mould Resistance

Proteins potentially inhibitory to the growth of grain‐moulding fungi in vitro have been identified from sorghum seeds. However, their role in vivo during fungi infection is still not clear. The objective of the present study was to evaluate the presence of antifungal proteins (AFPs) during grain development. Sureño (a grain mould‐resistant line), RTx430 (a grain mould‐susceptible) and their F₁ hybrid, were planted at two moisture levels. Caryopses were collected from each genotype every 7 days after anthesis (DAA) during development and maturity of the grain. Significant levels of grain mould occurred naturally. Levels of four AFPs (sormatin, chitinases, β‐1,3‐glucanases and ribosomal‐inactivating proteins) were determined using the immunoblotting technique. During grain development (7–35 DAA), Sureño and the F₁ hybrid showed higher levels of sormatin and chitinase than RTx430. RIPs levels in Sureño and the F₁ hybrid were higher than those in RTx430 after 21 DAA. Sormatin, chitinases, β‐1,3‐glucanases and RIPs levels in Sureño and in the F₁ hybrid were higher than those of RTx430 after grain physiological maturity. AFPs are associated with grain mould resistance because Sureño and the F1 hybrid induce and/or retain higher AFPs levels under grain mould infection pressure than did RTx430.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.): II. DEVELOPMENT OF GERMINABILITY, VIGOUR, AND DESICCATION TOLERANCE

The germinability, vigour, and desiccation tolerance of muskmelon(Cucumis melo L. cv. Top Mark) seeds was studied in relationto changes in seed water content during development within thefleshy fruit. Seed water content (fresh weight basis) declinedfrom 91% to 42% between 10 d and 35 d after anthesis (DAA) (whenmaximum dry weight was attained), then declined more slowlyto a minimum of 35% at 50 DAA before increasing again to 43%at 65 DAA. Fresh intact seeds were first germinable at 25 to30 DAA and attained maximum germination percentages at 45 DAA.Between 15 and 35 DAA, cotyledons, hypocotyls, radicles andepicotyls of isolated embryos (testa and perisperm enveloperemoved) sequentially developed the ability to grow when incubatedon water. Dehydration to water contents less that those attainedwithin the fleshy fruit is not a requirement for developmentof germination capacity of muskmelon seeds. Seeds became tolerantof rapid desiccation after 25 DAA, and drying of immature seeds(25 to 40 DAA) increased their germination percentages uponsubsequent imbibition. Washing, drying, or washing followedby drying increased seedling vigour (root length) as comparedto fresh seeds, which had very poor vigour. Water absorptionisotherms were constructed to test whether changes in water-bindingcomponents were correlated with the development of desiccationtolerance. Isotherms for seeds older than 25 DAA fit well tothe D'Arcy/Watt model, which postulates the existence of high-affinity,low-affinity and multi-molecular water-binding sites. Desiccation-intolerantseeds younger than 25 DAA lacked the component of the absorptionisotherm characteristic of the high-affinity water-binding siteswhich have been hypothesized to confer desiccation tolerance.However, we were unable to determine whether the absence ofhigh-affinity binding characteristics was specifically relatedto desiccation intolerance or was artifactual due to the lossof volatiles when immature seed samples were dried at high temperatures. Key words: Muskmelon, embryo, germination, development, vigour, desiccation     

Water Status Changes During Development in Relation to the Germination and Desiccation Tolerance of Aesculus hippocastanum L. Seeds

Seed growth characteristics of Aesculus hippocastanum were examinedin detail during development from about 70 to 140 d after anthesis(DAA), mainly in 1988 and 1989. Mean fresh and dry weights increasedfor both the axis and the whole seed up to the time of peakseed fall at 135 DAA with no cessation before fruit abscission.Water per seed increased up to 100 DAA, after which no furtherincrease occurred; moisture content declined for the embryonicaxis and whole seed respectively from above 75 and 65% at 95DAA to 65 and 50% at 130 DAA. At fruit shedding in 1990 waterpotential values of -1·2, -2·6 and -1·1MPa were observed for the testa, cotyledon and axis tissuesrespectively; relevant sorption isotherms are presented. Decreases in seed moisture content during development were accompaniedby increases in desiccation tolerance and in germinability,both reaching their maximum at the time of peak seed fall. Atmaturity, only about 10% viability was retained on drying seedto 20% moisture content; it is confirmed that the seeds are'recalcitrant'. The exact relationship between moisture contentand germination during development was dependent on the deptof dormancy, as judged by the period of chilling required; eachduration of chilling at 2°C within the range 3-12 weeksyields a curve of sigmoid shape. No germination occurred at26°C without chilling, but nearly full germination can beobserved for samples collected at 6 weeks before maximum seedfall with 12 weeks chilling. The rate of moisture loss duringdesiccation at 15°C and 15% rh becomes reduced during development.The ontogeny of these 'recalcitrant' seeds is compared withthat of 'orthodox' seeded species and the implication of sigmoid-shapedcurves for the relationship between seed moisture content andgermination are considered.Copyright 1993, 1999 Academic Press Aesculus hippocastanum L., horse chestnut, seed development, water status, germination, desiccation intolerance, desiccation rate     

The levels of soluble nucleotides in wheat aleurone tissue

The content of soluble nucleotides in aleurone layers isolated from mature wheat (Triticum aestivum var. Olympic) grain was investigated. The most abundant nucleotides were adenosine triphosphate, uridine triphosphate, and uridine diphosphoglucose. Smaller amounts of guanosine triphosphate, cytidine triphosphate, adenosine diphosphate, and nicotinamide adenine dinucleotide were also identified. The levels of some of these nucleotides were increased after incubation of the tissue under certain conditions.     

Ultrastructural Modifications of Plastids and Starch Metabolism during the Microsporogenesis of Ophrys lutea (Orchidaceae)

Ultrastructure of plastids has been studied throughout microsporogenesisof Ophrys lutea Cav. (Orchidaceae). A typical dedifferentiation/redifferentiationcycle of plastids was observed. At prophase I, plastids of pre-meiocytesgradually lost starch and separated the meiocytes through cytomicticchannels. At meiosis, plastids dedifferentiated to protoplastsand redifferentiated in young microspores. During pollen grainformation and maturation a cycle of starch accumulation/degradationoccurred; the mature pollen grain is starchless. Results arediscussed in terms of genetic inheritance following haploidnuclear state and in terms of carbohydrate metabolism. Plastids, starch, microsporogenesis, ultrastructure, Ophrys lutea, Orchidaceae     

Enzymes of carbohydrate metabolism in the developing rice grain

The levels of reducing and nonreducing sugars, starch, soluble protein, and selected enzymes involved in the metabolism of sucrose, glucose-1-P, and glucose nucleotides were assayed in dehulled developing rice grains (Oryza sativa L. line IR1541-76-3) during the first 3 weeks after flowering. The level of reducing sugars in the grain was highest 5 to 6 days after flowering. The level of nonreducing sugars and the rate of starch accumulation were maximum 11 to 12 days after flowering, when the level of soluble protein was also the highest. The activities of bound and free invertase, sucrose-UDP and sucrose-ADP glucosyltransferases, hexokinase, phosphoglucomutase, nucleoside diphosphokinase, and UDP-glucose and ADP-glucose pyrophosphorylases were high throughout starch deposition, and were maximum, except for nucleoside diphosphokinase which did not increase in activity, between 8 and 18 days after flowering. Soluble primed phosphorylase and ADP glucose-α-glucosyltransferase (starch synthetase) were both present during starch accumulation. Phosphorylase activity was at least 2-fold that of soluble starch synthetase but the synthetase followed more closely the rate of starch accumulation in the grain. The activity of starch synthetase bound to the starch granule also increased progressively with increased starch content of the grain.     

Adenylate and nicotinamide nucleotides in developing soybean seeds during seed-fill

Profiles of adenylate and nicotinamide nucleotides in soybean seeds were determined during seed-fill. The ATP content per seed increased during the early seed-filling stages to a level of 10 to 12 micrograms per seed. Seed ATP decreased after 40 days of development and reached its lowest level of less than 1 microgram at maturity. The ATP:ADP ratios were relatively constant at all seed development stages. Sharp increases in AMP levels during the late seed-fill stages were paralleled with a disappearance of ATP and ADP pools resulting in a reduced seed energy charge. Energy charge varied from the highest value of 0.78 at mid-seed-fill to less than 0.10 at maturity.     

Influence of Soil and Air Temperature on I. Nucleotide Metabolism and Growth of Phaseolus vulgaris L. seedlings

Eight day old greenhouse grown Phaseolus vulgaris L. seedlingswere exposed to three different temperature regimes. The concentrationof the total free nucleotides, nucleoside triphosphates, RNAand protein was much more pronounced in seedlings exposed tothe interactive day/night temperature of 28 ?C (soil) and 15?C (air) of regime I. Seedlings treated to a low soil/air, day/nighttemperature of 15 ?C (regime II) had the lowest content of totalfree nucleotides, nucleoside triphosphates, RNA and protein.The nucleoside triphosphate content and energy charge valueof seedlings treated to a high soil/air, day/night temperatureof 28 ?C (regime III) declined much earlier than in the seedlingsexposed to temperature regimes I and II. The fresh and dry weightof seedlings exposed to temperature regime I was significantlyhigher than those seedlings exposed to the other two temperatureregimes. The slowest growth was observed in seedlings exposedto temperature regime II. The noticeably higher concentrationof ATP, pyridine nucleotides and root nodulation in seedlingsexposed to temperature regime I suggests that higher soil andlow air temperature enhances nitrogen metabolism in P. vulgarisseedlings.     

The Number and Sizes of Starch Granules in the Wheat Endosperm, and their Association with Grain Weight

The number and size of starch granules was measured in developingand mature endosperms of two wheat varieties, Chinese Springand Spica. Chromosomal effects on particular aspects of starchgranule size and number were detected in the analysis of disomicgenotypes derived from reciprocal monosomic hybrids betweenthe two parental varieties. Among these genotypes, a greaterweight per endosperm cell was associated with a greater volumeper A-type starch granule. It is suggested that the number percell and volume of these large starch granules are the majordeterminants of endosperm cell weight, and there exists separategenetic control of these parameters. It should therefore bepossible genetically to combine these attributes to achievedirected increases in mature grain weight. Triticum aestivum, grain weight, starch granules