Biochemical changes in developing seeds of pigeonpea (Cajanus cajan)

Soluble sugars, starch, soluble nitrogen and protein nitrogen were studied in developing seeds of 3 cultivars of pigeonpea. When expressed on a per seed basis soluble sugars increased up to 35 days after flowering and then declined slightly. Rapid starch accumulation was observed between 14 and 28 days after flowering. The levels of soluble nitrogen and protein nitrogen underwent rapid changes during the same period. Amino-acid composition of seed protein was also studied at different stages of maturation. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of salt-soluble proteins revealed that seed storage globulins are formed after 14 days of flowering and do not change much during later stages of maturation.     

Changes in protein and carbohydrate content during development of seeds and siliquas of rapeseed (Brassica napus L.)

Abstract

A study was made on the changes observed in the protein, starch and soluble sugar content during development of siliquas and seeds of rapeseed grown in central Italy.

Concentration of starch and soluble sugars in the seed increases to 75 per cent dry matter during the first few weeks of pod development and then drops to minimum values. The protein increases steadily until maturity, when a level of 0.85 mg per seed is reached, equivalent to 18 per cent dry matter. The protein and starch in the hull? decrease continuously during development, while in the initial stages the soluble sugars are accumulated until they account for 33 per cent dry matter, after which they decline towards maturity.     

Relationships between indices of seed maturity and carrot seed quality

Carrot seeds cvs Chantenay and Amsterdam were harvested on several occasions from crops grown in 1985 and 1986, from the time they had reached or were close to their maximum dry weight and were starting to turn brown. Maximum seed dry weight occurred approximately 40–45 days after flowering (DAF) in both cultivars. Maximum germination (International Seed Testing Association 14-day count) occurred 40 and 55 DAF in cvs Chantenay and Amsterdam, respectively, but the maximum 7-day count and the minimum coefficient of variation of embryo length did not occur until 60 DAF in cv. Chantenay and 55 to 65 DAF in cv. Amsterdam. Percentage germination was negatively and linearly related to seed moisture content, chlorophyll a + b content in the seed coat and seed distortion, measured on a modified tensile-testing machine, the relationships accounting for 77, 71 and 64% of the variance in the 7-day germination count, respectively. The corresponding values for the 14-day count were 63, 61 and 50%. A simple field test for monitoring seed maturity is proposed.     

Lipid biosynthesis in seeds of mustard (Brassica juncea) influenced by zinc and sulphur deficiency

In developing seeds of mustard ( Brassica juncea L. cv. RLM 198) the period between 20 and 30 days after fertilization (DAF) was identified as the period of active lipid biosynthesis, although dry matter continued to accumulate until maturity. The period of lipid synthesis was associated with a decrease in starch, soluble sugars and protein, thus, giving rise to precursors for the biosynthesis of lipids. Besides decreasing the dry matter content (on both % and seed basis), Zn and S deficiency caused a significant ( P > 0.05) reduction in oil content. As compared to control, the decrease in oil content was 11, 12 and 18% at 30 DAF and 4, 9 and 16% at maturity in Zn, S and (Zn+S) deficient treatments, respectively. Throughout the period of seed development, a significant decrease in starch and protein with a slight accumulation of soluble sugars was observed due to deficiency of Zn or S. The rate of [l-¹⁴C]-acetate incorporation into total lipids, which was maximal at 30 DAF, also displayed a significant decrease due to the abovementioned mineral deficiencies. Addition of Zn or S in vitro, enhanced the lipid synthesis at all stages of seed development. Under Zn and S deficiency, the phospholipids increased from 10 to 30 DAF and then declined until maturity. However, the proportion of glycolipids and free fatty acids increased, with a corresponding decrease in total glycerides. Further, in deficiency treatments, there was an increase in 22:1 with a corresponding decrease in 18:1, 18:2 and 18:3 in developing and mature mustard seeds.     

Proteins of Soybean Seeds: II. Accumulation of the Major Protein Components during Seed Development and Maturation

Fresh weight and dry weight as well as quantitative and qualitative protein changes in the developing soybean (Glycine max) seed were described from 12 days after flowering until maturity. The seed proteins were separated on sucrose density gradients into three major fractions, having average sedimentation coefficients of 2.2S, 7.5S, and 11.8S. The 2.2S sedimenting proteins predominated at very early stages of development (12 days after flowering) and decreased proportionately throughout maturation. The 7.5S and 11.8S components appeared to be synthesized later in maturity and in larger amounts than the 2.2S proteins. Electrophoretic studies on extracts from whole seeds and on isolated protein fractions confirmed the early abundance of proteins in the 2.2S fraction and revealed temporal differences in the accumulation of three components of the 7.5S fraction. The 11.8S sedimenting fraction appeared throughout seed development as a homogeneous protein which accumulated in the seed with a time course similar to that of the total 7.5S protein fraction.     

Embryo-specific reduction of ADP-Glc pyrophosphorylase leads to an inhibition of starch synthesis and a delay in oil accumulation in developing seeds of oilseed rape

In oil-storing Brassica napus (rape) seeds, starch deposition occurs only transiently in the early stages of development, and starch is absent from mature seeds. This work investigates the influence of a reduction of ADP-Glc pyrophosphorylase (AGPase) on storage metabolism in these seeds. To manipulate the activity of AGPase in a seed-specific manner, a cDNA encoding the small subunit of AGPase was expressed in the sense or antisense orientation under the control of an embryo-specific thioesterase promoter. Lines were selected showing an embryo-specific decrease in AGPase due to antisense and cosuppression at different stages of development. At early developmental stages (25 days after flowering), a 50% decrease in AGPase activity was accompanied by similar decreases in starch content and the rate of starch synthesis measured by injecting (14)C-Suc into seeds in planta. In parallel to inhibition of starch synthesis, the level of ADP-Glc decreased, whereas Glc 1-phosphate levels increased, providing biochemical evidence that inhibition of starch synthesis was due to repression of AGPase. At 25 days after flowering, repression of starch synthesis also led to a decrease in the rate of (14)C-Suc degradation and its further metabolism via other metabolic pathways. This was not accompanied by an increase in the levels of soluble sugars, indicating that Suc import was inhibited in parallel. Flux through glycolysis, the activities of hexokinase, and inorganic pyrophosphate-dependent phosphofructokinase, and the adenylate energy state (ATP to ADP ratio) of the transgenic seeds decreased, indicating inhibition of glycolysis and respiration compared to wild type. This was accompanied by a marked decrease in the rate of storage lipid (triacylglycerol) synthesis and in the fatty acid content of seeds. In mature seeds, glycolytic enzyme activities, metabolite levels, and ATP levels remained unchanged, and the fatty acid content was only marginally lower compared to wild type, indicating that the influence of AGPase on carbon metabolism and oil accumulation was largely compensated for in the later stages of seed development. Results indicate that AGPase exerts high control over starch synthesis at early stages of seed development where it is involved in establishing the sink activity of the embryo and the onset of oil accumulation.     

Etude de la globuline des graines de Tournesol (Helianthus annuus L.) accumulation au cours de la maturation et localisation intracellulaire

Protein synthesis and accumulation in growing sunflower (Helianthus annuus L.cv.Airelle) seeds are studied. The salt soluble fraction, globulin, is the main soluble protein component. The earlier stages of seed development (10 days after flowering) are characterized by high M_r polypeptides (74, 58 and 44 kDa). Later stages mainly show nature globulin polypeptides. Thus, protein synthesis in seed occurs at a specific period of seed development which follows a period of fast cell divisions (0–14 days after flowering). Protein bodies are isolated and their protein composition analyzed. Globulin subunits are the main polypeptides of protein bodies soluble fraction. Mature globulin is only stored in protein bodies.      

Phosphorus metabolism during ripening ofGlycine max. (L.)Merril

The seeds, seed covers and leaves, taken after 17, 27, 37, and 47 days after tagging of flowers of soybean, were analysed quantitatively for their contents of different phosphorus fractions. Total phosphorus content increased in seed cover and leaves, there was a gradual decrease during ripening. All the phosphorus fractions i.e. acid soluble—P, lipid—P, nucleic acid—P, and protein—P were found to increase with maturity in seeds whereas in case of seed covers the content of acid soluble—P, nucleic acid—P and protein—P decreased but a marked increase was observed in lipid—P. In leaves during ripening, all the phosphorus fractions decreased except protein—P which was found to be almost constant. In lipid—P an increase was observed during later stages of maturity.     

Arabidopsis sucrose synthase 2 and 3 modulate metabolic homeostasis and direct carbon towards starch synthesis in developing seeds

Two genes encoding sucrose synthase (SUS), namely SUS2 (At5g49190) and SUS3 (At4g02280), are strongly and differentially expressed in Arabidopsis seed. Detailed biochemical analysis was carried out in developing seeds 9–21 days after flowering (DAF) of wild type and two knockouts. SUS2 and SUS3 are not redundant genes since single knockouts show a phenotype in developing seeds. The mutants had 30–50% less SUS activity and therefore accumulated 40% more sucrose and 50% less fructose at 15 DAF. This did not affect the hexose-P pool, but led to 30–70% less starch in embryo and seed coat. Lipids were 55% higher in both mutants at 9–15 DAF. It seems that sucrolysis via SUS is not required for oil or protein synthesis but rather for channeling carbon toward ADP-glucose and starch in seeds. Metabolite profiling with GC–TOF revealed specific downstream changes in primary metabolism as a consequence of signaling or regulatory fine-tuning. While sucrose increased, hexoses and specific amino acids decreased reciprocally. There was a developmental shift regarding an earlier timing of dry weight accumulation, germinative maturity, oil deposition, sugar levels, transient starch buildup, and protein storage. Nevertheless, final seed size and composition were unaltered due to an earlier cessation of growth, thus giving rise to an apparent silent phenotype of mature mutant seeds. We conclude that SUS is important for metabolite homeostasis and timing of seed development, and propose that an altered sucrose/hexose ratio can modify carbon partitioning and the pattern of storage compounds in Arabidopsis.     

Effect of nitrogen nutrition on endosperm protein synthesis in wild and cultivated barley grown in spike culture

In normal growth conditions, total protein percent (salt soluble plus hordein fractions) in the endosperm at maturity in barley cultivar Hordeum vulgare L. cv `Ruth' was about 14%, whereas in an accession of wild barley, Hordeum spontaneum Koch line 297, it was about 28%. Spike culture experiments were conducted to ascertain whether there were basic differences between the two genotypes under conditions of widely different nitrogen supply. Spikes of each genotype were grown from 8 to 25 days after flowering in in vitro culture in a growth medium containing 0 to 4 grams per liter nitrogen supplied as NH<sub>4</sub>NO<sub>3</sub>. Spikes were pulse-labeled at intervals from 12 to 24 days after flowering with 3.7 megabecquerel of [<sup>3</sup>H]leucine to determine relative rates of synthesis of hordein-1 and hordein-2 polypeptides. At low nitrogen levels `Ruth' had a lower protein content than 297, but at increasing nitrogen levels its protein content increased rapidly and reached a maximum (35%) higher than 297 (30%). The relative contribution of the hordein fraction to total protein increased mainly with time, and hordein-1 to total hordein increased mainly with nitrogen level, in both genotypes. There appeared to be no fundamental limitations in the capacity of `Ruth' to accumulate protein; 297 appears to have a greater basal level of nitrogen availability under normal conditions.     

Nodulation,nitrogen fixation and nitrogen uptake in pigeonpea (Cajanus cajan (L.) Millsp) of different maturity groups

Summary The seasonal patterns of nodulation, acetylene reduction, nitrogen uptake and nitrogen fixation were studies for 11 pigeonpea cultivars belonging to different maturity groups grown on an Alfisol at ICRISAT Center, Patancheru, India. In all cultivars the nodule number and mass increased to a maximum around 60–80 days after sowing and then declined. The nodule number and mass of medium- and late-maturing cultivars was greater than that of early-maturing cultivars. The nitrogenase activity per plant increased to 60 days after sowing and declined thereafter, with little activity at 100 days when the crop was flowering. At later stages of plant growth nodules formed down to 90 cm below the soil surface but those at greater depth appeared less active than those near the surface. All the 11 cultivars continued to accumulate dry matter until 140 days, with most biomass production by the late-maturing cultivars (up to 11 t ha⁻¹) and least by the early-maturing determinate cultivars (4 t ha⁻¹). Total nitrogen uptake ranged from 69 to 134 kg ha⁻¹. Nitrogen fixation by pigeonpea was estimated as the difference in total nitrogen uptake between pigeonpea and sorghum and could amount to 69 kg N ha⁻¹ per season, or half the total nitrogen uptake. Fixation by pigeonpea increased with crop duration, but there were differences within each maturity group. The limitations of the methods used for estimating N₂ fixation by pigeonpea are discussed. Submitted as J.A. No. 552 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Biochemical changes during germination and seedling growth in Cuscuta campestris

Changes in the contents of starch, protein, DNA, RNA, total phosphorus, acid soluble phosphorus and inorganic phosphorus, and in the activities of some enzymes of carbohydrate, amino acid, nucleic acid and phosphate metabolism were studied during the germination of Cuscuta campestris seeds. The results are expressed on per seed basis.
Starch content in Cuscuta seeds showed a steady decline with most of it depleted by the end of the eighth day of germination. Protein content increased with germination up to 48 h and then decreased. RNA and DNA contents increased to a maximal level on the fourth day of germination and then decreased. Total phosphorus in the seeds remained almost unchanged during the period of study. Both trichloroacetic acid soluble and inorganic phosphorus increased until the third day and then decreased. Phytin was rapidly hydrolyzed with little being detectable by the seventh day of germination. Glucose-6-phosphate dehydrogenase increased with germination, while fructose bisphosphate aldolase which is indispensable for glycolysis, decreased with germination. Ribonuclease and deoxyribonuclease increased till the third and fourth day, respectively, and then decreased. Aspartate and alanine aminotransferases showed a maximum on the second day and then decreased. Activities of alkaline fructose-1,6-bisphosphatase and phytase were absent in the dry seeds and appeared only on the second day of germination. Both α- and β-amylase activities were present in the dry seed.     

烯效唑干拌种对小麦氮素积累和运转及籽粒蛋白质品质的影响

通过田间试验,研究了不同烯效唑干拌种剂量对3个不同筋力小麦品种植株氮素积累、运转和籽粒蛋白质品质的影响,结果表明,基因型、环境及烯效唑处理对小麦品质的影响效应依次减小,且均达到了极显著水平,但三者的互作效应较小。烯效唑处理后提高了不同生态点下不同小麦品种籽粒蛋白质含量和产量,处理后的面筋含量和沉淀值增加,面团形成时间和稳定时间延长;干拌种增加了开花期各营养器官中的氮素含量和单株氮素积累量,花后氮素总转移量、总转移率及其对籽粒氮的贡献率极显著提高,且处理后旗叶中可溶性蛋白质含量在花后15 d内均显著高于对照;对籽粒中氮含量而言,烯效唑处理后提高了灌浆初期籽粒中的非蛋白氮含量,花后5—20 d内均高于对照,灌浆期间籽粒蛋白氮含量均高于对照,因而处理后的粗蛋白质含量变化动态特点为谷底高、回升快。研究认为,烯效唑处理如同基因、环境一样独立影响小麦籽粒品质,而烯效唑处理后提高了开花初期旗叶中的可溶性蛋白质含量和花前营养器官中氮素含量及花后氮素转运量,可能是其提高籽粒非蛋白氮含量、促进籽粒蛋白质含量增加和蛋白质质量提高的重要原因之一,烯效唑干拌种对小麦籽粒蛋白质品质的改善具有广适性。     

The influence of nutrition on the nitrogenous constituents of plants

Summary A pot experiment has been made with oats fertilized with increasing amounts of calcium nitrate, phosphorus, and various combinations of these. The yield of dry matter and the content of phosphorus, potassium, total nitrogen, soluble nitrogen, peptide nitrogen, and nitrate nitrogen were determined on three separate occasions during the growing period.The number of tillers was increased to a certain extent by nitrogen and then decreased again with further applications. Phosphorus alone had no influence, but a positive interaction between phosphorus and nitrogen on the number of tillers was observed. Nitrogen increased the number of seeds per spikelet and the percentage of grain.Except at maturity the content of potassium followed the content of total nitrogen, the percentage of which increased with increasing nitrogen supply. The content of nitrate nitrogen decreased during the growing period and at maturity was decreased by increasing phosphorus supply.As a percentage of the total nitrogen, the peptide nitrogen and soluble non-nitrate nitrogen in the straw decreased with increasing nitrogen supply. Peptide nitrogen increased at maturity. When the phosphorus was added with a heavy dressing of calcium nitrate, the percentage and the yield of protein nitrogen seemed to increase.     

Protein Bodies of Developing Seeds of Vicia faba

Changes in fine structure and starch, nitrogen, and solublesugar content were followed through to maturation in developingcotyledons of Vicia faba. Various ultrastructural changes wereobserved in the developing cotyledons, notably an increase inthe number of membrane-bound ribosomes which corresponded withthe onset of storage protein deposition. The build-up of storageprotein was shown to occur in the cytoplasm within membrane-boundvacuoles which subsequently became the protein bodies of themature seed, retaining the original tonoplast as the boundingmembrane of the protein body. Nuclei became lobed during thelater phases of maturation; phytoferritin was observed in plastidsof mature seeds. The deposition of reserves in the cotyledonswas complete by 85–90 days after flowering, followingwhich water was lost until the seed became hard and ‘ripe’by no days after flowering.     

Protein Synthesis in Cotyledons of Pisum sativum L: I. Changes in Cell-Free Amino Acid Incorporation Capacity during Seed Development and Maturation

The changes in protein content of pea cotyledons have been followed during the period from 9 to 33 days after flowering. Initially protein content increased gradually with a rapid period of deposition occurring between days 21 and 27 after flowering. After the 28th day the rate of accumulation of protein declined as the seed dehydrated and matured. At maturity the pea cotyledon contained approximately 25% protein which was divided into albumins and globulins in the ratio of 1:1.4.     

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 ).     

苦瓜果实发育过程中的主要营养成分变化及种子生活力演进

为明确苦瓜的果实和种子的适宜采收时期,采用"翠中翠"苦瓜为研究对象,对苦瓜果实发育过程中的可溶性蛋白、总可溶性糖、维生素C的含量及种子生活力的变化进行动态监测。结果显示:(1)花后第14天至第26天,随着果实的迅速膨大,可溶性蛋白、总可溶性糖及维生素C含量也迅速增长;(2)种子干物质迅速积累,含水量迅速下降,种子生活力迅速提高,至花后第22天至第26天,种子生活力基本稳定。     

Changes in oligosaccharide content and antioxidant enzyme activities in developing bean seeds as related to acquisition of drying tolerance and seed quality.

Seeds of bean (Phaseolus vulgaris cv. Vernel) were collected throughout their development on the plant and dried at 15 degrees C and 75% relative humidity to a final moisture content of about 16% (fresh weight basis) to determine whether the onset of tolerance to this drying condition was related to changes in soluble sugars or the activities of the main antioxidant enzymes, namely superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR). Measurements of soluble sugars and enzyme activities were made after drying the seeds, and drying tolerance was evaluated by the ability of dried seeds to germinate and to produce normal seedlings. Seeds became tolerant to drying at 45 d after anthesis, a time marking physiological maturity. At physiological maturity, the moisture content of seeds was about 50-55% (fresh weight basis) and seed dry matter reached about 190 mg per seed. Seed vigour, evaluated by controlled deterioration and conductivity measurements, continued to increase after seed mass maturity, but decreased when seeds remained thereafter for more than 7 d on the plant. Acquisition of drying tolerance was coincident with an accumulation of raffinose and stachyose. Dried-tolerant seeds were also characterized by a high amount of sucrose, the most abundant sugar, and by a low content of monosaccharides. The (raffinose+stachyose)/sucrose ratio increased during seed filling, reaching a value close to 1 when all the seeds became tolerant to drying, and maintaining this proportion during the final stages of maturation. Acquisition of drying tolerance was also related to a reorientation of the enzymatic antioxidant defence system. Drying-tolerant dried seeds displayed high CAT and GR activities and low SOD and APX activities, while the opposite condition was observed in immature dried seeds. The shift in antioxidant enzymes corresponded to the beginning of the maturation-drying phase. These results suggest that oligosaccharide metabolism and enzymatic antioxidant defences may be involved in acquisition of drying tolerance during bean seed development, but are not related to seed vigour.