Seed coat cell turgor in chickpea is independent of changes in plant and pod water potential

Turgor pressure in cells of the pod wall and the seed coat of chickpea (Cicer arietinum L.) were measured directly with a pressure probe on intact plants under initially dry soil conditions, and after the plants were irrigated. The turgor pressure in cells of the pod wall was initially 0.25 MPa, and began to increase within a few minutes of irrigation. By 2-4 h after irrigation, pod wall cell turgor had increased to 0.97 MPa. This increase in turgor was matched closely by increases in the total water potential of both the pod and the stem, as measured by a pressure chamber. However, turgor pressure in cells of the seed coat was relatively low (0.10 MPa) and was essentially unchanged up to 24 h after irrigation (0.13 MPa). These data demonstrate that water exchange is relatively efficient throughout most of the plant body, but not between the pod and the seed. Since both the pod and the seed coat are vascularized tissues of maternal origin, this indicates that at least for chickpea, isolation of the water relations of the embryo from the maternal plant does not depend on the absence of vascular or symplastic connections between the embryo and the maternal plant.     

Structure and development of Medicago truncatula pod wall and seed coat

BACKGROUND AND AIMS: Medicago truncatula has gained much attention as a genomic model species for legume biology, but little is known about the morphology of its pods and seeds. Structural and developmental characteristics of M. truncatula pod walls and seed coats are presented. METHODS: Plants of M. truncatula ecotype A17 were grown under controlled conditions in a greenhouse. Flowers were date-tagged at anthesis, so that pods of known age could be collected. Harvested pods were fixed and sectioned for light microscopy. Structural attributes of pod walls and seed coats were characterized at four time points throughout early to mid-stages of pod development (3, 6, 13 and 20 d post-pollination). KEY RESULTS: Basic features of the pod wall are an exocarp comprised of a single epidermal layer, a mesocarp with seven to 14 layers of parenchyma cells, and an endocarp composed of an inner epidermal cell layer and three to five layers of sclerenchyma cells adjacent to it. Vascular bundles are abundant in the pod wall and include one lateral carpellary bundle, one median carpellary bundle and nine to 12 vascular bundles, all embedded within the mesocarp parenchyma. Seed coat features include an epidermal layer of macrosclereids, a sub-epidermal layer of osteosclereids, and two to five rows of internal parenchyma cells. The hilar region contains the tracheid bar and the chalazal vascular bundle, the latter of which expands to form only two short branches. CONCLUSIONS: This characterization provides a needed understanding of pod structure and development in this model legume, and should facilitate various molecular investigations into legume fruit and seed biology.     

Gas exchange by pods and subtending leaves and internal recycling of CO(2) by pods of chickpea (Cicer arietinum L.) subjected to water deficits

Terminal drought markedly reduces leaf photosynthesis of chickpea (Cicer arietinum L.) during seed filling. A study was initiated to determine whether photosynthesis and internal recycling of CO(2) by the pods can compensate for the low rate of photosynthesis in leaves under water deficits. The influence of water deficits on the rates of photosynthesis and transpiration of pods and subtending leaves in chickpea (cv. Sona) was investigated in two naturally-lit, temperature-controlled glasshouses. At values of photosynthetically active radiation (PAR) of 900 micromol m(-2) s(-1) and higher, the rate of net photosynthesis of subtending leaves of 10-d-old pods was 24 and 6 micromol m(-2) s(-1) in the well-watered (WW) and water-stressed (WS) plants when the covered-leaf water potential (Psi) was -0.6 and -1.4 MPa, respectively. Leaf photosynthesis further decreased to 4.5 and 0.5 micromol m(-2) s(-1) as Psi decreased to -2.3 and -3.3 MPa, respectively. At 900--1500 micromol m(-2) s(-1) PAR, the net photosynthetic rate of 10-d-old pods was 0.9-1.0 micromol m(-2) s(-1) in the WW plants and was -0.1 to -0.8 micromol m(-2) s(-1) in the WS plants. The photosynthetic rates of both pods and subtending leaves decreased with age, but the rate of transpiration of the pods increased with age. The rates of respiration and net photosynthesis inside the pods were estimated by measuring the changes in the internal concentration of CO(2) of covered and uncovered pods during the day. Both the WW and WS pods had similar values of internal net photosynthesis, but the WS pods showed significantly higher rates of respiration suggesting that the WS pods had higher gross photosynthetic rates than the WW pods, particularly in the late afternoon. When (13)CO(2) was injected into the gas space inside the pod, nearly 80% of the labelled carbon 24 h after injection was observed in the pod wall in both the WW and WS plants. After 144 h the proportion of (13)C in the seed had increased from 19% to 32% in both treatments. The results suggest that internal recycling of CO(2) inside the pod may assist in maintaining seed filling in water-stressed chickpea.     

Seed coat cell turgor responds rapidly to air humidity in chickpea and faba bean

The turgor pressure in cells of chickpea (Cicer arietinum L.) and faba bean (Vicia faba L.) seed coats was measured with a pressure probe. Measurements were made under in situ conditions by removing a section of wall from a pod, which remained attached to the plant, and exposing the intact seed. If the pod wall was removed and the turgor measurements made under ambient laboratory conditions of 50% to 70% relative humidity (RH), cell turgor pressure declined over time, typically reaching 0 MPa. If the pod wall was removed and the turgor measurements made under conditions of 100% RH, however, cell turgor pressure was stable over time, relatively uniform within the seed coat tissue, and was found to be 0.1-0.3 MPa for chickpea, and 0.1-0.2 MPa for faba bean. In both species there was a marked decline in cell turgor, beginning within about 60 s, when humidification was discontinued. The decline in cell turgor occurred regardless of the depth of the cell within the seed coat tissue, and this decline could be stopped, but not entirely reversed, when humidification was restored. An increase in cell turgor could also be caused by wetting of the seed. These responses indicate that a very rapid water exchange can occur within the seed coat tissue in situ. The rapid and, in some cases, relatively permanent loss of seed coat cell turgor in the absence of humidification raises serious concerns regarding desiccation artefacts which may be involved in the empty seed coat technique, often used to study seed carbon and water relations in grain legumes.     

Seed coat development, anatomy and scanning electron microscopy of Harpagophytum procumbens (Devil's Claw), Pedaliaceae

Seed coat development of Harpagophytum procumbens (Devil's Claw) and the possible role of the mature seed coat in seed dormancy were studied by light microscopy (LM), transmission electron microscopy (TEM) and environmental scanning electron microscopy (ESEM). Very young ovules of H. procumbens have a single thick integument consisting of densely packed thin-walled parenchyma cells that are uniform in shape and size. During later developmental stages the parenchyma cells differentiate into 4 different zones. Zone 1 is the multi-layered inner epidermis of the single integument that eventually develops into a tough impenetrable covering that tightly encloses the embryo. The inner epidermis is delineated on the inside by a few layers of collapsed remnant endosperm cell wall layers and on the outside by remnant cell wall layers of zone 2, also called the middle layer. Together with the inner epidermis these remnant cell wall layers from collapsed cells may contribute towards seed coat impermeability. Zone 2 underneath the inner epidermis consists of large thin-walled parenchyma cells. Zone 3 is the sub-epidermal layers underneath the outer epidermis referred to as a hypodermis and zone 4 is the single outer seed coat epidermal layer. Both zones 3 and 4 develop unusual secondary wall thickenings. The primary cell walls of the outer epidermis and hypodermis disintegrated during the final stages of seed maturation, leaving only a scaffold of these secondary cell wall thickenings. In the mature seed coat the outer fibrillar seed coat consists of the outer epidermis and hypodermis and separates easily to reveal the dense, smooth inner epidermis of the seed coat. Outer epidermal and hypodermal wall thickenings develop over primary pit fields and arise from the deposition of secondary cell wall material in the form of alternative electron dense and electron lucent layers. ESEM studies showed that the outer epidermal and hypodermal seed coat layers are exceptionally hygroscopic. At 100% relative humidity within the ESEM chamber, drops of water readily condense on the seed surface and react in various ways with the seed coat components, resulting in the swelling and expansion of the wall thickenings. The flexible fibrous outer seed coat epidermis and hypodermis may enhance soil seed contact and retention of water, while the inner seed coat epidermis maintains structural and perhaps chemical seed dormancy due to the possible presence of inhibitors.     

Ontogenic changes in enzymes of carbon metabolism in relation to carbohydrate status in developing mungbean reproductive structures

The content of free sugars and the activities of enzymes involved in carbon metabolism-sucrose synthase, acid and alkaline invertase, phosphoenol pyruvate carboxylase, malic enzyme and isocitrate dehydrogenase were determined during seed development in mungbean pods. A decrease in carbohydrate content of pod wall from 10 to 25 days after flowering (DAF) and a concomitant increase in the seed till 20 DAF was observed. Sucrose remained the dominant soluble sugar in the pod wall and seed. In the branch of inflorescence and pod wall, the activities of sucrose metabolizing enzymes, viz. acid and alkaline invertase, sucrose synthase (synthesis and cleavage) and sucrose phosphate synthase were higher at 5-10 DAF, whereas in seed the maximum activities of these enzymes were observed at the time of maximum seed filling stage (10-20 DAF). High activities of sucrose synthase at the time of rapid seed filling can be correlated to its sink strength. Higher activities of phosphoenol pyruvate carboxylase in the branch of inflorescence and pod wall than in seed may indicate the involvement of the fruiting structure for recapturing respired CO2. High activities of isocitrate dehydrogenase and malic enzyme in the seed at the time of rapid seed filling could provide NADPH and carbon skeletons required for the synthesis of various seed reserves.     

Response of sugar interconversion, starch and protein accumulation to supplied metabolites during pod filling in chickpea

Detached chickpea inflorescences bearing pods at 20 days after flowering (DAF) were cultured for 5 days in complete liquid medium supplemented separately with asparate, myo-inositol, alpha-ketoglutarate and phytic acid. Effect of these metabolites on sugar interconvestion and starch and protein accumulation in developing pods was studied. Substituting asparate (62.5 mM) for glutamine in culture medium decreased relative proportion of sucrose in all pod tissues but increased the level of sugars, starch and protein in pod wall and cotyledons. In cotyledons, whereas myo-inositol (75 mM) reduced the accumulation of starch without affecting protein level, alpha-ketoglutarate (44 mM) increased both starch and protein accumulation. Both myo-inositol and alpha-ketoglutarate increased relative proportion of sucrose in cotyledons. Phytic acid (1 mM) decreased in cotyledons 14C incorporation from glucose into EtOH extract (principally constituted by sugars), amino acids and proteins but increased the same into starch. In cotyledons, phytic acid also increased 14C incorporation from glutamate into amino acids but this increase was negatively correlated with protein synthesis. Phytic acid decreased the relative distribution of 14C from glucose and glutamate into sucrose from pod wall but enhanced the same into EtOH extract from embryo. Based on the results, it is suggested that mode of metabolic response to exogenously supplied metabolites widely differs in pod tissues of chickpea.     

Embryo development in the lady's slipper orchid, Paphiopedilum delenatii, with emphasis on the ultrastructure of the suspensor

BACKGROUND AND AIMS: Owing to large-scale collecting, the lady's slipper orchid, Paphiopedilum delenatii, is under threat of extinction. Asymbiotic germination provides a useful way to re-establish plants in the wild and for commercial propagation. A detailed study of embryo development would provide information on subsequent germination events and aid in the propagation of the species. METHODS: Developing capsules were collected for histochemical and ultrastructural studies by using both light and transmission electron microscopy. KEY RESULTS: The suspensor of this species consists of three vacuolated cells. During the early globular stage of embryo development, structural differentiation occurs, revealing an abundance of smooth endoplasmic reticulum cisternae and wall ingrowths within the suspensor cells. These features are not present in cells of the embryo proper. Furthermore, the results of Nile red staining demonstrate that a cuticular layer is present only in the embryo proper, but absent from the suspensor. Cuticular material is also present in the inner walls of the seed coat, and persists through seed maturation. CONCLUSIONS: The morphological features of the transfer cell and the absence of cuticular material in the suspensor cell wall corroborate the hypothesis that the suspensor is the major nutrient uptake site for the developing embryo in the lady's slipper orchid. The absence of an endosperm and presence of cuticular material in the inner walls of the seed coat enclosing the embryo proper further support the notion that nutrient uptake by the embryo is confined to the micropylar end of the seed through the suspensor.     

Histological Study of Seed Coat Development in Arabidopsis thaliana

Arabidopsis seed coat development using light and transmission electron microscopy revealed major morphological changes associated with the transition of the integuments into the mature seed coat. By the use of a metachromatic staining procedure, cytological events such as the production of phenolic compounds and acidic polysaccharides were followed. Immediately after fertilization, the cells of the inner epidermis of the inner integument became vacuolated and subsequently accumulated pigment within them. This pigment started to disappear from the cytoplasm at the torpedo stage of the embryo, as it became green. During the torpedo stage, mucilage began to accumulate in the cells of the external epidermis of the outer integument. Furthermore, starch grains accumulated against the central part of the inner periclinal wall of these cells, resulting in the formation of small pyramidal domes that persisted until seed maturity. At the maturation stage, when the embryo became dormant and colourless, a new pigment accumulation was observed in an amorphous layer derived from remnants of crushed integument layers. This second pigment layer was responsible for the brown seed colour. These results show that seed coat formation may proceed in a coordinated way with the developmental phases of embryogenesis. Received 25 May 1999/ Accepted in revised form 10 February 2000     

Formation of the vascular system of developing bean (Phaseolus limensis L.) seeds according to nuclear magnetic resonance microtomography

1H magnetic resonance microtomography imaging was applied to study vascular systems in developing bean (Phaseolus limensis L.) seeds. Using the gradient echo method, we recorded 2D tomographic sections in the sagittal and axial planes of the fruits sampled from a vegetating plant on days 10, 17, 24, and 31 after fertilization. Any vascular connection between the tissues of maternal plant (bean pod and seed coat) and the embryo were undetectable. The embryo has an autonomous branched network of procambial strands in the cotyledons, converging to the embryonic axis. The bean pods are covered with a network of vascular bundles; large vascular strands run along the dorsal and ventral sutures. The seed coat vascular bundles are formed in the process of seed ripening and are represented by a developed vascular system multiply branching in the middle part of the ground parenchyma at the stage of physiological maturity. They are connected with the source of assimilates via the lateral pod veins and a large vascular bundle, entering the seed below the hilum via the placenta. Assimilates enter the external part of the seed coat, which contains no vascular bundles, via the funiculus vascular bundles and hilum tissue.     

Differential response of carbon and nitrogen metabolism to fluoride application in fruiting structures of chickpea (Cicer arietinum)

The effect of sodium fluoride (10 and 50 mol·m⁻³) on the activities of sucrose metabolizing enzymes, transaminases and glutamine synthetase in relation to the transformation of free sugars to starch and protein in the fruiting structures (pod wall, seed coat, cotyledons) of chickpea was studied by culturing detached reproductive shoots in a liquid medium. Addition of fluoride to the culture medium drastically reduced starch content of the cotyledons and caused a marked build-up of total free sugars comprised mainly of reducing sugars in the pod wall and seed coat, and sucrose in the cotyledons. Concomitantly, the activity of soluble invertase was stimulated in the pod wall but reduced in the cotyledons. However, soluble protein content of both the pod wall and the cotyledons increased in conjunction with an increase in the activities of glutamate-oxaloacetate transaminase, glutamate-pyruvate transaminase and glutamine synthetase. Disruption of starch biosynthesis under the influence of fluoride and the resulting accumulation of free sugars possibly resulted in their favoured utilization in nitrogen metabolism. Labelling studies with [U-¹⁴C]-sucrose showed that the ¹⁴C incorporation into total free sugars was enhanced by fluoride in the pod wall but reduced in the seed coat and cotyledons, possibly due to an inhibitory effect on their translocation to the developing seeds.     

Ultrastructure of the “fringe-layer”, the innermost epidermis of cotton seed coats

Summary The inner epidermis of the inner integument of cotton seed coats (fringe-layer) and the cuticles between this cell layer and the nucellus were examined in the light and electron microscope at different times of their development. The cells of the fringe-layer contain only small vacuoles and their cytoplasm is densely packed with organelles and free and membrane-bound polysomes. The lateral walls contain many plasmodesmata. At the time when the fruit capsules stop growing, the fringe-cells produce a cell wall labyrinth, resembling that of transfer cells. The cell wall labyrinth is restricted to the lateral walls. The differentiated state of the fringe-cells is short-lived. At about the time of elaboration of the cell wall labyrinth most of them become progressively vacuolated, lignify, and lose their cytoplasmic constituents. The development of the fringe-layer is well correlated with other developmental events in the inner integument, but not with the filling of embryo and endosperm with reserve substances.At anthesis, the fringe-layer and nucellus are covered by a thin cuticle proper of about 20 nm. After anthesis, the nucellar cells start to produce a cuticular layer of considerable, but variable, thickness (0.25–2.5 m), containing a polysaccharide network.In drying seeds the cells of the fringe-layer disrupt. The thin outer tangential wall remains attached to the seed coat. The rest of the cell, together with the cuticles and the collapsed cells of the nucellus, form a protective layer around embryo and endosperm, remaining attached to the seed coat at the chalazal end.     

Relationships between Respiration Rate and Adenylate and Carbohydrate Pools of the Soybean Fruit

Relationships between respiration rate and adenylate and carbohydrate pools of the soybean (Glycine max L. Merrill) fruit during rapid seed growth were evaluated. Plants at mid pod-fill were subjected to different concentrations of CO(2) to alter the amount of photosynthate produced and, thus, available to the fruit. Respiration rate of the intact fruits was measured, along with glucose, sucrose, and starch concentrations, adenylate energy charge (AEC), and total adenylate pool (SigmaAdN) in the pod wall, seed coat, and cotyledons. The concentration of sucrose remained relatively constant in the pod wall (1.0 milligram per 100 milligrams dry weight), seed coat (6.5 milligrams per 100 milligrams dry weight), and cotyledons (4.5 milligrams per 100 milligrams dry weight) at moderate and high respiration rates. Furthermore, AEC remained relatively constant in the pod wall (0.55), seed coat (0.24), and cotyledons (0.44) during changes in respiration rate. This suggests that the amount of assimilate transported to the fruit, and its flux through the sucrose pools of the fruit parts, were important in the regulation of the respiration rate of the fruit. The average SigmaAdN in the seed coat (1300 picomoles per milligram dry weight) was significantly greater than in the cotyledons (750 picomoles per milligram dry weight) and pod wall (300 picomoles per milligram dry weight). In addition, the SigmaAdN in the seed coat and cotyledons increased with increasing respiration rate of the fruit. The high SigmaAdN in the seed coat and its increase with increases in respiration rate of the fruit suggest that an energy-requiring process is involved in the movement of sucrose through the seed coat.     

Discrimination Against 13CO2 in Leaves,Pod Walls,and Seeds of Water-stressed Chickpea

The rate of photosynthesis (P _N) in leaves and pods as well as carbon isotope content in leaves, pod walls, and seeds was measured in well-watered (WW) and water-stressed (WS) chickpea plants. The P _N, on an area basis, was negligible in pods compared to leaves and was reduced by water stress (by 26%) only in leaves. WS pod walls and seeds discriminated less against ¹³CO₂ than did the controls. This response was not observed for leaves as is usually the case. Pod walls and seeds discriminated less against ¹³CO₂ than did leaves in both WW and WS plants. Measurement of carbon isotope composition in pods may be a more sensitive tool for assessing the impact of water stress on long-term assimilation than is the instantaneous measurement of gas exchange rates.     

Formation of the vascular system of developing bean (<Emphasis Type="Italic">Phaseolus limensis</Emphasis> L.) seeds according to nuclear magnetic resonance microtomography

¹H magnetic resonance microtomography imaging was applied to study vascular systems in developing bean (Phaseolus limensis L.) seeds. Using the gradient echo method, we recorded 2D tomographic sections in the sagittal and axial planes of the fruits sampled from a vegetating plant on days 10, 17, 24, and 31 after fertilization. Any vascular connection between the tissues of maternal plant (bean pod and seed coat) and the embryo were undetectable. The embryo has an autonomous branched network of procambial strands in the cotyledons, converging to the embryonic axis. The bean pods are covered with a network of vascular bundles; large vascular strands run along the dorsal and ventral sutures. The seed coat vascular bundles are formed in the process of seed ripening and are represented by a developed vascular system multiply branching in the middle part of the ground parenchyma at the stage of physiological maturity. They are connected with the source of assimilates via the lateral pod veins and a large vascular bundle, entering the seed below the hilum via the placenta. Assimilates enter the external part of the seed coat, which contains no vascular bundles, via the funiculus vascular bundles and hilum tissue.     

In Vivo Fixation of CO2 by Attached Pods of Brassica campestris L.

In vivo net CO₂ exchange characteristics of attached Brassicapods were studied during the entire period of their growth anddevelopment after anthesis. ¹⁴CO₂ was fed both from the externalatmosphere and internally through the pod cavity, and the anatomyof the pod-wall was examined microscopically. Stomata were observedin the outer epidermal layer of the pod wall. Net in vivo CO₂fixation by the pods was observed throughout the period of theirdevelopment and was maximum on day 42 after anthesis (DAA).Compared to the internal feeding experiments, ¹⁴CO₂ fixationfrom the external environment was very high. Apparent translocationof fixed carbon from the pod wall to seeds was rapid. Pod photosynthesiscontributed substantially to seed growth. pods, Brassica campestris L., CO₂ fixation, stomata     

In Vivo Fixation of CO2 by Attached Pods of Brassica campestris L.

In vivo net CO₂ exchange characteristics of attached Brassicapods were studied during the entire period of their growth anddevelopment after anthesis. ¹⁴CO₂ was fed both from the externalatmosphere and internally through the pod cavity, and the anatomyof the pod-wall was examined microscopically. Stomata were observedin the outer epidermal layer of the pod wall. Net in vivo CO₂fixation by the pods was observed throughout the period of theirdevelopment and was maximum on day 42 after anthesis (DAA).Compared to the internal feeding experiments, ¹⁴CO₂ fixationfrom the external environment was very high. Apparent translocationof fixed carbon from the pod wall to seeds was rapid. Pod photosynthesiscontributed substantially to seed growth. pods, Brassica campestris L, CO₂ fixation, stomata     

Regulation of monocarpic senescence of Brassica campestris by the developing pods

Senescence of Brassica campestris L. cv. B-9 was studied with regard to seed maturation and source-sink relationships. In normal control plants leaf senescence (as determined by the change in chlorophyll level) started and proceeded in a progressive manner from base to apex during the period of early pod setting. Complete yellowing of the leaves occurred well before the seed maturation and pod wall senescence. The pod wall always senesced before the attainment of final seed weight. In two different sets of acrocarpous plants containing 65 pods and 10 pods, respectively, leaf senescence was delayed during the pod filling period. It started non-sequentially after complete yellowing and senescence of the pod wall. The degree of leaf senescence at the post-pod filling stage was almost proportional to the number of pods present. When peduncles of the acrocarpous 10-podded plants were removed after the pod filling stage of the plant, leaf senescence was delayed compared to plants whose pedicels were removed, although the senescence pattern of the upper three leaves was nonsequential in both cases. Defruiting at an early stage of development delayed leaf senescence, although the pattern of such senescence remained unaltered (i.e. nonsequential). Defoliation hastened the seed-filling process and pod wall senescence. Plants containing fewer pods had higher average seed weight, although yield per plant was reduced.
These results suggest that the pod wall serves as a temporary as well as intermediary storage organ and that foliar senescence is not directly related to seed maturation. The possible cause of uncoupling between foliar senescence and seed development is discussed.     

砂仁种子的解剖学和组织化学研究

砂仁种子包括假种皮、种皮、外胚乳、内胚乳与胚。假种皮由内表皮、外表皮及其间的６－９层薄壁细胞组成。种皮分为外种皮、中种皮与内种皮。外种皮由１层表皮细胞构成,其壁增厚并略木质化。中种皮包括各含１层细胞的下层皮和半透明细胞层与含３－５层细胞的色素层;下皮层与色素层细胞均含有红综色素,后者的壁呈网状增厚。内种皮由１层内切向壁与径向壁非常增厚的石细胞构成。种皮表面具有许多疣状突起,它们是体积较小的表皮细胞     

Biochemical and Physiological Changes During Maturation of Fruit of the Field Pea (Pisum arvense L.)

Quantitative changes in the carbohydrates, soluble nitrogenouscompounds, and specific proteins of seeds and pods of fieldpea have been followed during two seasons and related to theoverall pattern of growth of the embryo and other parts of thefruit. Reserves of solutes are established in the pod and inthe seed coat and endosperm of the seed some time before theembryo commences its exponential phase of growth. Later, thesereserves disappear and it is estimated that their mobilizationmight provide, at the most, only one-fifth of the embryo's requirementsfor carbon and nitrogen. Starch and certain albumin-type proteins accumulate relativelyearly in the life of the embryo. Globulin-type proteins, bycontrast, are laid down only after the cotyledons are almostfull size and after most of the reserve of starch has been accumulated.The content of hemicellulose, the principal carbohydrate ofthe embryo, increases steadily as the embryo grows. Neitherhemicellulose nor globulin is found in appreciable amounts inorgans other than the cotyledons. Each organ of the fruit has its own highly characteristic poolof free amino acids. Orderly and reproducible changes occurin the composition of these pools during fruit development,suggesting that variations in the amounts of specific compoundsmay have special significance in the programme of developmentand synthesis in the fruit.