Changes in nitrogen contents and in proteolytic activities in different parts of field-grown wheat ears (Triticum aestivum L.) during maturation

Aminopeptidase, carboxypeptidase and neutral endopeptidase activitieswere analyzed in glumes and in kernels of field-grown wheat(Triticum aestivum L.) during ear development. Kernels harvestedon two dates were subdivided into outer pericarp, cross cells,endosperm and embryo. In developing parts with a net nitrogeninflux (young glumes, embryo, endosperm) the aminopeptidaseactivity is high, but in nitrogen-mobilizing tissues (senescingglumes, Outer pericarp) this activity decreases. Carboxypeptidaseis most active in fully expanded tissues. Neutral endopeptidaseshows the highest activity in the nitrogen mobilizing partsand extremely low activity in the embryo and the endosperm. (Received July 15, 1978; )     

Ploidy barrier to endosperm development in maize

Maize kernels inheriting the indeterminate gametophyte mutant (ig) on the female side had endosperms that ranged in ploidy level from diploid (2x) to nonaploid (9x). In crosses with diploid males, only kernels of the triploid endosperm class developed normally. Kernels of the tetraploid endosperm class were half-sized but with well-developed embryos that regularly germinated. Kernels of endosperm composition other than triploid or tetraploid were abortive.-Endosperm ploidy level resulting from mating ig/ig x tetraploid Ig similarly was variable. Most endosperms started to degenerate soon after pollination and remained in an arrested state. Hexaploid endosperm was exceptional; it developed normally during the sequence of stages studied and accounted for plump kernels on mature ears. Since such kernels have diploid maternal tissues (pericarp) but triploid embryos, the present finding favors the view that endosperm failure or success in such circumstances is governed by conditions within the endosperm itself.-Whereas tetraploid endosperm consisting of three maternal genomes and one paternal genome is slightly reduced in size but supports viable seed development, that endosperm having two maternal and two paternal chromosome sets was highly defective and conditioned abortion. Thus, development of maize endosperm evidently is affected by the parental source of its sets of chromosomes.     

The germination and the forces required to penetrate seed layers of different seedlots of three cultivars of crisp lettuce

Using 24 commercial seedlots of the lettuce cvs Ithaca, Pennlake and Saladin, the percentage germination at a range of temperatures from 20 to 35°C, and the forces required to penetrate the pericarp, the endosperm and the embryo were measured. Percentage germination declined with increasing temperature in all seedlots but there were particularly large differences between seedlots of the same cultivar at 30.0 and 32.5°C. The force required to penetrate the pericarp was on average more than twice that required to penetrate the endosperm. It is concluded that in commercial cultivars of crisp lettuce the strength of the pericarp plays a much greater role than that of the endosperm in determining germination. In all three cultivars there were significant positive correlations between seed weight and the force required to penetrate the whole seed. However only in cv. Pennlake were there significant correlations between germination at high temperature and seed penetration forces.     

A yellow mosaic disease of horse chestnut (Aesculus spp.) caused by apple mosaic virus

A severe foliar yellow mosaic disease was observed in horse chestnut trees (Aesculus carnea and A. hippocastanum). Reactions in woody indicator plants grafted with diseased horse chestnut suggested the presence of an ilarvirus. Virus isolates obtained by mechanical inoculation of herbaceous test plants reacted with antisera to apple mosaic virus but not with antisera to its serotype prunus necrotic ringspot virus, or to prune dwarf virus. Yellow mosaic was induced in horse chestnut seedlings grafted with tissues from herbaceous hosts infected with horse chestnut isolates or with the European plum line pattern isolate of apple mosaic virus. Virus was detected by enzyme-linked immunosorbent assay (ELISA) in embryo and endosperm of immature seed from infected trees but not in mature seed, or progeny seedlings. Strawberry latent ringspot virus was detected in one of six A. hippocastanum trees with a leaf vein yellows disease.     

rgf1, a mutation reducing grain filling in maize through effects on basal endosperm and pedicel development

The maize cob presents an excellent opportunity to screen visually for mutations affecting assimilate partitioning in the developing kernel. We have identified a defective kernel mutant termed rgf1, reduced grain filling, with a final grain weight 30% of the wild type. In contrast with most defective endosperm mutants, rgf1 shows gene dosage-dependent expression in the endosperm. rgf1 kernels possess a small endosperm incompletely filling the papery pericarp, but embryo development is unaffected and the seeds are viable. The mutation conditions defective pedicel development and greatly reduces expression of endosperm transfer layer-specific markers. rgf1 exhibits striking morphological similarities to the mn1 mutant, but maps to a locus approximately 4 cM away from mn1 on chromosome 2 of maize. Despite reduced starch accumulation in the mutant, no obvious lesion in starch biosynthesis has been detected. Free sugar levels are unaltered in rgf1 endosperm. Rates of sugar uptake, measured over short (8 h) periods in cultured kernels, are increased in rgf1 compared to the wild type. rgf1 and wild-type kernels, excised at 5 DAP and cultured in vitro also develop differently in response to variations in sugar regime: glucose concentrations above 1% arrest placentochalazal development of rgf1 kernels, but have no effect on cultured wild-type kernels. These findings suggest that either uptake or perception of sugar(s) in endosperm cells at 5-10 DAP determines the rgf1 kernel phenotype.     

Accrescimento del pericarpo,seme, endosperma ed embrione in prunus Amygdalus stokes

Abstract

GROWTH OF PERICARP, SEED, ENDOSPERM, AND EMBRYO IN PRUNUS AMYGDALUS STOKES. — The fruits of an almond-tree growing at Bari were collected weekly from February 22nd to July 11th and on August 16th 1960. The material was kept in fixative; the growth of the various organs was studied both from a morphological and a quantitative point of view. Special attention was given to growth of the endosperm, especially during the nuclear stage and at the beginning of cellularisation (Figg. 1-14), and to the developement of the embryo until it reaches the « heart-shaped » stage (Figg. 15–22). From a quantitative point of view, the volume and main diameters of pericarp and seed, and whenever possible endosperm and embryo, were measured for each fruit. Most of the data are given in Tables I to V and Figg. 23 and 24.

If reference is made to the 3 phases of fruit growth established for other species (notably peaches and cherries), the main conclusions are that:

1. phase I (growth of pericarp, testa and nucellus) is clearly recognisable; it ends after the micropylar portion of the endosperm has become cellular and the embryo heart shaped;

2. phase II is also present: during this phase most of the growth of endosperm and embryo takes place; while the seed has reached its definite size at the end of phase I, the pericarp undergoes a period of greatly reduced growth;

3. two weeks after the beginning of phase II the pericarp seems to resume growth just for a very short period, judging at least by the weekly values of the ratio pericarp volume to seed volume (see Fig. 23); this seems to indicate the existence of a new phase, that is phase III, which in fleshy fruits of the genus Prunus corresponds to a much longer and important process of pericarp growth than in the almond;

4. as in the peaches and cherries therefore a crisis in pericarp growth occurs during the period of maximum rate of growth of the cellular endosperm and embryo;

5. the sequence: cellularisation of the endosperm, growth of endosperm and embryo, ceasing of seed growth, and reduction in pericarp growth is very clear, particularly if we take into account growth in length rather than in volume; both morphological and quantitative data would indicate the importance of the endosperm not only for the beginning of embryo development, but also for the control of pericarp growth.

     

Pseudogamous Apomixis in Maize and Sorghum in Diploid-Tetraploid Crosses

Apomictic seed development is a complex process including formation of unreduced embryo sac, parthenogenetic embryo development from the egg cell, and endosperm formation either autonomously, or due to fertilization of polar nuclei by the sperm (under pseudogamous form of apomixis). In the latter case, an obstacle to the normal endosperm development is disturbance of maternal (m) -to-paternal (p) genomic ratio 2m: 1p that occurs in the cases of pollination of unreduced embryo sac with haploid sperms. Usage of tetraploid pollinators can overcome this problem because in such crosses maternal-to-paternal genomic ratio is 4m: 2p that provides formation of kernels with plump endosperm. Using tetraploid lines as pollen parents we observed formation of plump kernels on the ears and panicles of diploid maize and sorghum accessions. These kernels had hybrid endosperm and diploid maternaltype embryo or hybrid embryo with different ploidy level (2n, 3n, 4n). The frequencies of plump kernels on the ear ranged from 0.2-0.3% to 5.7-6.2% counting from the number of ovaries. Maternal-type plants were found in two maize lines, their frequency varying from 10.7 to 37.5% of the progeny plants. In CMS-lines of sorghum pollinated with tetraploid sorghum accessions, the frequency of plump kernels ranged from 0.6 to 14.0% counting from the number of ovaries; the frequency of maternal-type plants varied from 33.0 up to 96.1%. The hybrid nature of endosperm of the kernels that gave rise to maternal-type plants has been proved by marker gene expression and by SDS-electrophoresis of endosperm proteins. These data testify to variable modes of seed formation under diploid × tetraploid crosses in maize and sorghum both by amphi- and by apomixis. Therefore, usage of tetraploid pollinators might be a promising approach for isolation of apomixis in maize and sorghum accessions.     

Endosperm and Embryo Development in Daucus carota L.

Maximum carrot seed dry weight and maximum endosperm volumewere reached about 35 d after anthesis, although at this timethe endosperm was still soft, the pericarp green and less than50% of the seeds were viable. Fully viable, ripe seeds werenot produced until 44 d later. Seventy per cent of the increasein endosperm volume was due to an increase in cell number, whichceased 35 d after anthesis. The increase in embryo volume wasslower and was due to an increase in both cell number and cellvolume which continued until 49 d after anthesis. At maturitythe embryo was the equivalent of between 2% and 3% of the endospermvolume. The relationship between embryo length and cell number per embryowas unaffected by seed crop plant density, seed crop harvestdate and position of the seed on the mother plant but it wasaffected by the year of seed production, possibly due to differencesin temperature during the period of seed growth. Key words: Endosperm, Embryo, Carrot, Development     

Water uptake and splitting of wild oat caryopsis

Imbibition and germination experiments were conducted on the caryopses of wild oats (Avena fatua L.). The embryo envelopes, pericarp and aleurone layer, which completely cover the embryo-endosperm, do not form barriers against water uptake. The initial uptake of water is passive and the water moves across the pericarp with ease as it contains cracks; it is, however, transported across the aleurone layer through its cell walls into the endosperm and embryo of the caryopsis. The starchy endosperm enlarges due to water uptake causing the pericarp to rupture, thus exposing the aleuronelayer-covered seed. The aleurone layer is structurally heterogenous consistings of radially compressed irregular cells and cuboidal or radiallys tretched cells; the latter contains thicker walls. The former type is present along the abaxial side of the embryo and in the crease on the adaxial side of the caryopsis; the latter type covers the endosperm. The physical distention of the endosperm due to water uptake causes the rupture of the pericarp and the aleurone layer, and facilitates the emergence of the radicle and coleorhiza of the embryo during caryopsis germination.     

A Preliminary Study on Neutral and Acid Inhibitors in the Seed of Kalopanax scptemlobus

The method of extraction, isolation, purification and identification of the neutral and acid inhibitors in the seeds of Kalopanax Septemlobus was given. It is proved that the neutral inhibitor is coumarin, the acid inhibitor is abscisic acid (ABA) by means of paper chromatograph, thin layer chromatograph, high performance liquid chromatograph (HPLC), color reaction and bioassay. The neutral inhibitor can strongly inhibite not only the seed germination of Brassica Chinensis and the radical elongation, but also that of the seed of Kalopanax septemlobus. The study showed that ABA and coumarin exist in the seed coat, endosperm and pericarp of the Kalopanax Septemlobus seed. Both ABA and coumarin can transport from seed cover (pericarp, seed coat)to the interior (endosperm, embryo) as the seeds were stored in refrigerator. In addition, the different results of extracting neutral inhibitor with water, methanol, and alcohol were compared in this paper.     

刺楸种子中性与酸性抑制物质的研究

本文研究了刺楸种子抑制物质的提取,分离和鉴定方法,并测定了抑制物质的相对活性及其含量。结果表明,刺揪种子中除含高水平的酸性抑制物外,还存在一种中性抑制物。该中性抑制物同酸性抑制物相似,有较强的生物活性,不但可以抑制白菜种子的萌发与胚根生长,而且对已解除休眠的刺楸种子也有显著作用。利用纸层析,薄层层析,颜色反应和高效液相色谱等手段,证明中性抑制物质为香豆素类物质;酸性抑制物质的主要成分为脱落酸。种子各部位均含这两种抑制物质,但含量有所不同,随种子贮藏时间的延长,抑制物会发生转移。本文还讨论了用不同溶剂提取中性抑制物的效果。     

A mutational approach to the study of seed development in maize

The maize seed comprises two major compartments, the embryo and the endosperm, both originating from the double fertilization event. The embryogenetic process allows the formation of a well-differentiated embryonic axis, surrounded by a single massive cotyledon, the scutellum. The mature endosperm constitutes the bulk of the seed and comprises specific regions containing reserve proteins, complex carbohydrates, and oils. To gain more insight into molecular events that underlie seed development, three monogenic mutants were characterized, referred to as emp (empty pericarp) on the basis of their extreme endosperm reduction, first recognizable at about 12 d after pollination. Their histological analysis reveals a partial development of the endosperm domains as well as loss of adhesion between pedicel tissues and the basal transfer layer. In the endosperm, programmed cell death (PCD) is delayed. The embryo appears retarded in its growth, but not impaired in its morphogenesis. The mutants can be rescued by culturing immature embryos, even though the seedlings appear retarded in their growth. The analysis of seeds with discordant embryo-endosperm phenotype (mutant embryo, normal endosperm and vice-versa), obtained using B-A translocations, suggests that emp expression in the embryo is necessary, but not sufficient, for proper seed development. In all three mutants the picture emerging is one of a general delay in processes related to growth, as a result of a mutation affecting endosperm development as a primary event.     

Development of the Almond Nut (Prunus dulcis (Mill.) D. A. Webb). Anatomy and Chemical Composition of Fruit Parts from Anthesis to Maturity

The growth of the fruit of two varieties of almond (Prunus dulcis(Mill.) D. A. Webb) was studid from anthesis (week 0) to maturity(week 32). The dimensions, fresh weight, moisture content, anatomyand chemical composition of the pericarp, testa, embryo, endospermand nucellus are recorded diagrarnmatically, graphically andby micrographs for one variety. Of the two ovules present atflowering only one normally developed further. By 12 weeks afterflowering the whole fruit had reached full size. The space encloscdby the pericarp was filled by nuallus until weck 10, with subsequentenlargement of both endosperm and embryo. From week 16 to week20 the embryo increased to full size with a concumnt decreasein the size of the endosperm. Sixteen weeks after flowering,the embryo began to accumulate protein and lipid, little ofwhich originated from either the nucellus or endosperm. Theembryo contained no starch or reducing sugar but up to 3% sucrosein the early stags which dtcreascd as lipid and protein increased.Starch and sucrose levels were high in the testa at week 16but subsbquently dropped, starch more rapidly than sucrose.The role of the testa in transport of metabolites to the embryois discussed. Prunus dulcis, almond, fruit development, anatomy, embryo, endosperm     

Enzymes of sucrose and hexose metabolism in developing kernels of two inbreds of maize

Tissue distribution and activity of enzymes involved in sucrose and hexose metabolism were examined in kernels of two inbreds of maize (Zea mays L.) at progressive stages of development. Levels of sugars and starch were also quantitated throughout development. Enzyme activities studied were: ATP-linked fructokinase, UTP-linked fructokinase, ATP-linked glucokinase, sucrose synthase, UDP-Glc pyrophosphorylase, UDP-Glc dehydrogenase, PPi-linked phosphofructokinase, ATP-linked phosphofructokinase, NAD-dependent sorbitol dehydrogenase, NADP-dependent 6-P-gluconate dehydrogenase, NADP-dependent Glc-6-P dehydrogenase, aldolase, phosphoglucoisomerase, and phosphoglucomutase. Distribution of invertase activity was examined histochemically. Hexokinase and ATP-linked phosphofructokinase activities were the lowest among these enzymes and it is likely that these enzymes may regulate the utilization of sucrose in developing maize kernels. Most of the hexokinase activity was found in the endosperm, but the embryo had high activity on a dry weight basis. The endosperm, which stores primarily starch, contained high PPi-linked phosphofructokinase and low ATP-linked phosphofructokinase activities, whereas the embryo, which stores primarily lipids, had much higher ATP-linked phosphofructokinase activity than did the endosperm. It is suggested that PPi required by UDP-Glc pyrophosphorylase and PPi-linked phosphofructokinase in the endosperm may be supplied by starch synthesis. Sorbitol dehydrogenase activity was largely restricted to the endosperm, whereas 6-P-gluconate and Glc-6-P dehydrogenase activities were highest in the base and pericarp. A possible metabolic pathway by which sucrose is converted into starch is proposed.     

Distribution of enzyme activities within the developing maize (Zea mays) kernel in relation to starch, oil and protein accumulation

The association of enzyme activities in developing kernels with specific storage product accumulation at maturity was analyzed in different parts of Zea mays inbred OH43 kernels. Maize kernels were harvested at 20 and 55 days post-pollination and dissected into basal region, pericarp, embryo, lower endosperm, middle endosperm and upper endosperm. Mature (55 days pos(-pollination) kernel parts were analyzed for starch, total protein, zein and oil content. Immature (20 days post-pollination) kernel parts were assayed for activities of 15 enzymes of sugar and amino acid metabolism. Statistical analyses of the data suggested that glucokinase (EC 2.7.1.2) fructokinase (EC 2.7.1.4) and phosphofructokinase (EC 2.7.1.1 11) activities were primarily associated with oil accumulation, whereas ADP'-glueose pyrophosphorylasc (EC 2.7.7.27) and sucrose synthase (EC 2.4.1.13) activities were associated with starch accumulation. The results suggest that oil biosynthesis utilizes inveitase-mediated sucrose degradation in a pathway not requiring pyrophosphatc. whereas starch biosynthesis utilizes a sucrose synthase-mediated pathway of sucrose degradation in a pathway requiring pyrophosphatc. Additional groups of enzyme activities were associated with each oilier but not with any specific storage product and appeared to be associated with general metabolic activity.     

Changes in the Strength of Lettuce Endosperm during Germination

The forces required to puncture intact lettuce (Lactuca sativa) seed and pericarp, endosperm and embryo were measured by the Instron Universal Testing Machine. It required about 0.6 newton to puncture the endosperm in seeds imbibed in the dark at 6, 12 and 24 hours. Endosperm of seeds imbibed in the light or in dark with gibberellic acid required about 4.2 newtons at 6 and at 12 hours and only about 0.15 newton at 24 hours. Forces required to puncture embryo at all treatments and times remained constant at about 0.3 newton. Changes in the strength of the endosperm do not appear to be related directly to protrusion of the radicle.     

Tissue distribution and developmental patterns of NADH-dependent and ferredoxin-dependent glutamate synthase activities in maize (Zea mays) kernels

Both NADH-dependent glutamate synthase (NADH-GOGAT, EC 1.4.1.14) and ferredoxin-dependent glutamate synthase (Fd-GOGAT, EC 1.4.7.1) activities were present in the endosperm, embryo, pedicel and pericarp of maize ( Zea mays L. var. W64A × A619) kernels. The endosperm contained the highest proportions of each activity on a per tissue basis. In the endosperm, NADH-GOGAT and Fd-GOGAT activities increased 12- and 2.5-fold, respectively, during early zein accumulation. NADH-GOGAT and Fd-GOGAT activities were expressed in the upper, middle and lower portions of the endosperm in a manner that paralleled but preceded zein accumulation. Maize endosperm NADH-GOGAT was purified 159-fold using ammonium sulfate fractionation, anion exchange chromatography and dye-ligand chromatography. Apparent K_m values for glutamine, α-ketoglutarate and NADH were 850, 19 and 1 μM, respectively. The results are consistent with endosperm GOGAT functioning to redistribute nitrogen from glutamine, the predominant nitrogenous compound delivered to the endosperm, into other amino acids needed for storage protein synthesis.