油莎豆的组织培养和快速繁殖

1植物名称油莎豆(Cyperus esculentus L.var.sativus Boeck),又名油莎草、铁荸荠、地下板栗、地下核桃、人参果和人参豆。2材料类别块茎顶芽。3培养条件芽诱导培养基:(1)MS NAA1mg·L-1     

Chufa (Cyperus esculentus L. var. sativus boeck.): An unconventional crop. studies related to applications and cultivation

Tubers ofCyperus esculentuswere an important food in ancient Egypt. In Spain the tubers are used to make a beverage named horchata, and are also consumed as fresh after soaking. In other countries tubers are used in sweetmeats or uncooked as a side dish. New products obtained can enhance the interest in this crop: as a source of dietary fiber in food technology; as a high quality cooking/salad oil or as biodiesel fuel; as caramel to add body, flavor or color to other products; as a source of starch; as an antioxidant-containing food, etc. The results of a twenty-year research program on nutrition, fertilization, lodging control, planting date, soil texture and use of herbicides are shown. Three cultivars (‘Ametlla Bonrepos’, ‘Gegant Africana’, and ‘Llargueta Alboraia’) are selected and characterized using horticultural and morphological traits, the chemical composition of tubers and horchatas, and the RAPD technique.     

A molecular survey concerning the origin of Cyperus esculentus (Cyperaceae,Poales): two sides of the same coin (weed vs. crop)

Background and AimsCyperus esculentus is widespread in tropical and temperate zones and is also present in cooler regions. It is used as a crop plant, but it also occurs in the wild and as a weed. As a consequence of its ecological plasticity, C. esculentus has remarkable variability, with several morphotypes. Four wild-type varieties are presently recognized, in addition to the cultivated form. This study investigates the phylogenetic position and biogeography of C. esculentus with the objective of contributing new data to increase the understanding of its evolutionary history.Methods Genealogical relationships among genotypes were inferred by using plastid DNA haplotype and nuclear ribosomal (nr) DNA ribotype sequences for 70 specimens either collected in the field or obtained from herbaria. Statistical dispersal–vicariance (S-DIVA) and Bayesian binary method (BBM) analyses were used to reconstruct the possible ancestral ranges of C. esculentus. In order to determine the age of C. esculentus, a time-measured phylogenetic analysis was performed.Key Results Considerable variation between the chosen nuclear and plastid markers was detected (27 ribotypes vs. six haplotypes). No geographical structure was displayed among the haplotypes, but information on the dispersal pattern may be deduced. Two types of ribotypes were detected in nrDNA, with an evident geographical segregation into an Old World group and a polymorphic New World group. Both S-DIVA and BBM analyses suggested a biogeographical history in which dispersal from the African region has been crucial in shaping the current distribution pattern of C. esculentus. The most recent common ancestor between C. esculentus races has an age of 5.1 million years (95 % highest posterior density 2.5–10.2).Conclusions The molecular analysis provides novel insights into the evolutionary history of C. esculentus. The results have various taxonomic and phylogenetic implications, including a hypothesis on the origin and phylogeography of this species, which probably originated in the late Cenozoic in Africa, and reached the Americas repeatedly, independently of Columbian exchanges.     

Inheritance of seed weight in Cucumis sativus (L.) var. sativus and var. hardwickii (Royle) Kitamura

Summary A series of experiments was conducted to determine the inheritance of seed weight in cucumber. Matings between a Cucumis sativus var. sativus (Cs) L. inbred line (USDA WI 1606; P₁) and a C. sativus var. hardwickii (Royle) Kitamura (Ch) collection (PI 215589; P₂) were made to produce seed of reciprocal F₁, F₂, and BC₁ families. Families were grown under field and greenhouse conditions, and seeds were extracted from fruit 55 to 60 days post-pollination. Seed of F₁ and F₂ families was obtained using the Cs inbred WI2808 (P₁₂) and the Ch collection LJ 90430 (P₁₀), and seed of F₁ families were produced using a North Carolina Design II mating scheme in which three Cs (P₃= GY-14; P₄=WI 1379; P₅=WI 1909) inbreds were used as maternal parents and seven Ch collections (P₂; P₆= PI462369; P₇=486336; P₈=LJ91176; P₉=273469; P₁₀= 2590430; P₁₁=PI187367) were used as paternal parents. Mean seed weights of F₁ progeny reflected the dominance of genes of the C. sativus var. sativus parent. Transformation to number of seeds per unit weight resulted in increased variance homogeneity within generations and a broad-sense heritability ranging between 26% to 56%. Additive and dominance effects were important in the expression of seed weight in P₁×P₂ progeny produced in the greenhouse and additive effects were important in field grown progeny resulting from P₁×P₂ and P₁₀×P₁₂ matings. The estimated number of factors or loci involved ranged between 10 to 13, depending on the method of calculation.     

Multiple origins of congested inflorescences in Cyperus s.s. (Cyperaceae): Developmental and structural evidence

? Premise of the study: The understanding of homoplasic structures becomes more relevant when they are complex and define large angiosperm taxa. Inflorescence architecture usually fulfills both features, as happens with Cyperus, a genus with two taxonomical subdivisions characterized either by alternative expressions of Kranz anatomy (C(3) or C(4)) or inflorescence shape (condensed or lax). Those subdivisions are not completely congruent because at least one of these presumed characters has evolved several times. We focused a SEM study on the inflorescence development in species with condensed inflorescences and different photosynthetic anatomy to test the possibility that condensed inflorescences of subgen. Anosporum (C(3) anatomy) have evolved independently from those of subgen. Cyperus (C(4) anatomy). ? Methods: Freshly collected inflorescences of C. entrerianus, C. eragrostis, C. oxylepis, and C. incomtus were studied using stereoscopic and scanning electron microscopy. ? Key results: Condensed inflorescences of Cyperus species with C(3) and C(4) anatomy had differences in structure and development: (1) mature structure, (2) position of second-order branching initiation in the first developmental stage of the inflorescence, (3) main axis development and elongation, and branching development, (4) types of ramifications, (5) phyllotaxis and symmetry. ? Conclusions: Results support multiple origins of condensed inflorescences in Cyperus, based especially on differences in timing during development and elongation of the main axis and branches, branching pattern and phyllotaxis. Structure and development may be the key to using inflorescence morphology as an external feature to distinguish large natural groups within Cyperus based on vegetative anatomy.     

Safflower microsomes catalyse oil accumulation in vitro: A model system

Microsomal membrane preparations from the developing cotyledons of safflower (Carthamus tinctorius L.) seed catalyse the formation of triacylglycerol fromsn-glycerol 3-phosphate and linoleoyl-CoA. Conditions of incubation were achieved in which the rate of triacylglycerol synthesis approached activities which were compatible with oil accumulation observed in vivo. Reaction mixtures which contained the microsomes took on a white soup-like appearance as triacylglycerol synthesis proceeded and sufficient oil was produced to form a white fat-pad at the surface after centrifugation. The development of the oil bodies in the microsomal membranes was studied by electron microscopy and showed that lipid droplets were formed in or on the membrane surface and were then released as apparently naked entities into the surrounding medium. The ontogeny of the oil droplet in vitro is discussed in terms of oil-body formation in vivo.     

Towards a new classification of the giant paraphyletic genus Cyperus (Cyperaceae): phylogenetic relationships and generic delimitation in C4 Cyperus

Maximum likelihood and Bayesian inference analyses of nuclear ribosomal DNA (ETS1f) and plastid DNA (rpl32‐trnL, trnH‐psbA) sequence data are presented for ‘C₄ Cyperus’ (Cyperaceae). The term ‘C₄ Cyperus’ encompasses all species of Cyperus s.l. that use C₄ photosynthesis linked with chlorocyperoid vegetative anatomy. Sampling comprises 107 specimens of 104 different taxa, including many of the subdivisions of C₄ Cyperus s.s. and all C₄ segregate genera (Alinula, Ascolepis, Kyllinga, Lipocarpha, Pycreus, Queenslandiella, Remirea, Sphaerocyperus and Volkiella). According to our results, C₄ Cyperus is a well‐supported monophyletic clade nested in C₃ Cyperus. Despite the lack of resolution along the backbone of the C₄ Cyperus clade and for some internal branches, several well‐supported clades can be distinguished. The first clade in C₄ Cyperus is formed by Cyperus cuspidatus and C. waterloti. Other recognizable and well‐supported clades correspond to segregate genera, i.e. Ascolepis, Lipocarpha including Volkiella, and Kyllinga. Species of C₄ Cyperus s.s. form a core grade in which the C₄ segregate genera are embedded. Pycreus, the largest segregate genus composed of c. 120 species, is not monophyletic as it includes several C₄ species of Cyperus s.s. This study establishes a phylogenetic framework for revising the classification and character evolution in Cyperus s.l. © 2013 The Linnean Society of London     

A novel gene mutation that confers abnormal patterns of beta-carotene accumulation in cauliflower (Brassica oleracea var. botrytis)

The Or gene of cauliflower (Brassica oleracea var. botrytis) causes many tissues of the plant to accumulate carotenoids and turn orange, which is suggestive of a perturbation of the normal regulation of carotenogenesis. A series of experiments to explore the cellular basis of the carotenoid accumulation induced by the Or gene was completed. The Or gene causes obvious carotenoid accumulation in weakly or unpigmented tissues such as the curd, pith, leaf bases and shoot meristems, and cryptically in some cells of other organs, including the roots and developing fruits. The dominant carotenoid accumulated is beta-carotene, which can reach levels that are several hundred-fold higher than those in comparable wild-type tissues. The beta-carotene accumulates in plastids mainly as a component of massive, highly ordered sheets. The Or gene does not affect carotenoid composition of leaves, nor does it alter color and chromoplast appearance in flower petals. Interestingly, mRNA from carotenogenic and other isoprenoid biosynthetic genes upstream of the carotenoid pathway was detected both in orange tissues of the mutant, and in comparable unpigmented wild-type tissues. Thus the unpigmented wild-type tissues are likely to be competent to synthesize carotenoids, but this process is suppressed by an unidentified mechanism. Our results suggest that the Or gene may induce carotenoid accumulation by initiating the synthesis of a carotenoid deposition sink in the form of the large carotenoid-sequestering sheets.     

Inheritance of seed dormancy in Cummis sativus var. hardwickii (Royle) Alef.

Summary Reciprocal matings were made between two Cucumis sativus var. sativus L. inbred lines (WI 1606 and WI 2808) and two var. hardwickii (Royle) Alef. accessions (PI 215589 and PI 183967). Each case produced a series of reciprocal F₁, F₂, and BC₁ and BC₂ progenies which were used to evaluate seed dormancy in var. hardwickii. Under controlled conditions (25°±1°C and 85%±5% RH; 12 h fluorescent light, 30 mol s^–1 m^–2), no seed dormancy was observed in the var. sativus inbred lines 36 days following seed extraction from fruit. With rare exception, var. hardwickii accessions were dormant for at least 60 days. Seed dormancy in the F₁ was absent 36 days post extraction, indicating that dormancy in var. hardwickii is conditioned by recessive genes present. Seed of some F₁ progeny germinated between 36 and 50 days post-extraction, indicating the presence of transient dormancy or the more variable expression of the dormancy of var. hardwickii. No significant reciprocal differences in either germination rate or percentage were detected in either of the F₁ and F₂ progeny sets, suggesting lack of cytoplasmic or maternal control over these traits. It was estimated that three to seven factors or loci are involved with the expression of this trait depending on the method of calculation, and that a complex interaction between embryonic and non-embryonic tissue exists. Least square estimates indicate that both additive and dominance effects were important in the expression of dormancy. Comparison of theoretical geometric and arithmetic F₂ means to observed F₂ means also suggests that non-additive gene action contributes substantially to the observed variation. Broad-sense heritability ranged between 78 and 95%.     

Chemical investigation of different extracts and essential oil from the tubers of (Tunisian)Cyperus rotundus. Correlation with their antiradical and antimutagenic properties

The mutagenic potential of aqueous, Total Oligomers Flavonoids (TOF), ethyl acetate, and methanol extracts as well as essential oil (EO) obtained from tubers ofCyperus rotundus L. was assessed by “Ames assay”, usingSalmonella tester strains TA98 and TA100, and “SOS chromotest” usingEscherichia coli PQ37 strain with and without an exogenous metabolic activation system (S9). None of the different extracts showed a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test” and the “SOS chromotest”. Our results showed thatC. rotundus extracts have antimutagenic effects withSalmonella typhimurium TA98 and TA100 strains towards the mutagen Aflatoxin B1 (AFB1), as well as withE. coli PQ37 strain against AFB1 and nifuroxazide mutagens. A free radical scavenging test was used in order to explore the antioxidant capacity of the extracts obtained from the tubers ofC. rotundus. TOF, ethyl acetate and methanol extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. These extracts showed IC50 values of respectively 5, 20 and 65 μg/ml. The beneficial effects of TOF, ethyl acetate, methanol and essential oil extracts ofC. rotundus have been assessed by antioxidant and antimutagenic activities.     

The floral scales in Hellmuthia (Cyperaceae, Cyperoideae) and Paramapania (Cyperaceae, Mapanioideae): an ontogenetic study

BACKGROUND AND AIMS: In 1976 the monotypic genus Hellmuthia was placed in the Hypolytreae s.l., but was subsequently ascribed to the Mapanioideae, tribe Chrysitricheae, mainly because of the presence in Hellmuthia of two lateral, mapanioid-like floral scales with ciliated keels, the anatomy of the nutlet, the embryo and the inflorescence. Recently, based on cladistic analyses and supported by pollen ontogenetic evidence, Hellmuthia was transferred to a Cyperaceae, tribe Cypereae, clade mainly consisting of Ficinia and Isolepis. In this study, the floral ontogeny in Hellmuthia was investigated and compared with the floral ontogeny in Paramapania, with special attention for the floral scales. METHODS: Freshly collected inflorescences of Hellmuthia membranacea and Paramapania parvibractea were investigated using scanning electron and light microscopy. KEY RESULTS: In the conical 'spikelet' in Hellmuthia, proximal bracts occur, each axillating an axis with empty glume-like structures, or a reduced spikelet. Hence, it is a reduced partial inflorescence. In Hellmuthia, the stamen primordia originate before the primordia of the perianth-gynoecium appear. Moreover, a third adaxially positioned 'floral scale' was observed for the first time. The position and relative time of appearance of the floral scales in Hellmuthia are typical for perianth parts in Cyperoideae. The basal position of Hellmuthia within a clade of species with usually perianthless flowers, allows the presence of rudiments of a perianth in Hellmuthia to be interpreted as a primitive state. Development of the lateral 'scales' in Paramapania follows a different pattern. Therefore, it was decided that the lateral 'scales' in Paramapania are different from the lateral perianth parts in Hellmuthia. The pollen grains in Hellmuthia are cyperoid, with one polar and five lateral apertures, of which the membrane is covered with sexinous bodies. The pollen surface is granulate and perforate with microspines. CONCLUSIONS: The floral ontogeny in Hellmuthia occurs according to the general cyperoid pattern. The lateral scales in Hellmuthia are perianth parts, and they are not homologous to the lateral 'scales' in Paramapania.     

Application of Melatonin-Enhanced Tolerance to High-Temperature Stress in Cherry Radish (Raphanus sativus L. var. radculus pers)

Journal of Plant Growth Regulation - The growth and development of cold-season plants are susceptible to high temperature. Melatonin is a plant growth regulator with potential to improve plant...     

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.     

Distinct Phylogeographic Structures of Wild Radish (Raphanus sativus L. var. raphanistroides Makino) in Japan

Coastal plants with simple linear distribution ranges along coastlines provide a suitable system for improving our understanding of patterns of intra-specific distributional history and genetic variation. Due to the combination of high seed longevity and high dispersibility of seeds via seawater, we hypothesized that wild radish would poorly represent phylogeographic structure at the local scale. On the other hand, we also hypothesized that wild radish populations might be geographically differentiated, as has been exhibited by their considerable phenotypic variations along the islands of Japan. We conducted nuclear DNA microsatellite loci and chloroplast DNA haplotype analyses for 486 samples and 144 samples, respectively, from 18 populations to investigate the phylogeographic structure of wild radish in Japan. Cluster analysis supported the existence of differential genetic structures between the Ryukyu Islands and mainland Japan populations. A significant strong pattern of isolation by distance and significant evidence of a recent bottleneck were detected. The chloroplast marker analysis resulted in the generation of eight haplotypes, of which two haplotypes (A and B) were broadly distributed in most wild radish populations. High levels of variation in microsatellite loci were identified, whereas cpDNA displayed low levels of genetic diversity within populations. Our results indicate that the Kuroshio Current would have contributed to the sculpting of the phylogeographic structure by shaping genetic gaps between isolated populations. In addition, the Tokara Strait would have created a geographic barrier between the Ryukyu Islands and mainland Japan. Finally, extant habitat disturbances (coastal erosion), migration patterns (linear expansion), and geographic characteristics (small islands and sea currents) have influenced the expansion and historical population dynamics of wild radish. Our study is the first to record the robust phylogeographic structure in wild radish between the Ryukyu Islands and mainland Japan, and might provide new insight into the genetic differentiation of coastal plants across islands.     

The contribution of small collections to species distribution modelling: A case study from Fuireneae (Cyperaceae)

The recent and rapid digitization of biodiversity data from natural history collection (NHC) archives has enriched collections based data repositories; this data continues to inform studies of species' geographic distributions. Here we investigate the relative impact of plant data from small natural history collections (collections with < 100,000 specimens) on species distributional models in an effort to document the potential of data from small NHCs to contribute to and inform biodiversity research. We modelled suitable habitat of five test case species from Fuireneae (Cyperaceae) in the United States using specimen records available via the Global Biodiversity Information Facility and that of data ready to mobilize from two regional small herbaria. Data were partitioned into three datasets based on their source: 1) collections-based records from large NHCs accessed GBIF, 2) collections-based records from small NHCs accessed from GBIF, and 3) collections-based records from two small regional herbaria not yet mobilized to GBIF. We extracted and evaluated the ecological niche represented for each of the three datasets by applying dataset occurrences to 14 environmental factors, and we modelled habitat suitability using Maxent to compare the represented distribution of the environmental values among the datasets. Our analyses indicate that the data from small NHCs contributed unique information in both geographic and environmental space. When data from small collections were combined with data from large collections, species models of the ecological niche resulted in more refined predictions of habitat suitability, indicating that small collections can contribute unique occurrence data which enhance species distribution models by bridging geographic collection gaps and shifting modelled predictions of suitable habitat. Inclusion of specimen records from small collections in ongoing digitization efforts is essential for generating informed models of a species' niche and distribution.     

Fish emulsion as a food base for rhizobacteria promoting growth of radish (Raphanus sativus L. var. sativus) in a sandy soil

Commercial fish emulsion was evaluated as a plant growth medium and as a nutrient base to enhance radish (Raphanus sativus L. var. sativus) growth by bacterial and actinomycete isolates. Six bacterial isolates including three actinomycetes were selected from a screening of 54 bacteria (including 23 actinomycetes) based on their ability to produce plant growth regulators (PGRs) and to colonize radish roots. These isolates were tested in the presence and absence of autoclaved or non-autoclaved fish emulsion or inorganic fertilizers. The nutrient contents and types and levels of PGRs in tissues of treated plants were assayed to determine the basis of growth promotion. Fish emulsion was found to support plant growth in a sandy soil as effectively as an applied inorganic fertilizer. The plant growth promotion by bacterial and actinomycete isolates was most pronounced in the presence of autoclaved or non-autoclaved fish emulsion than in the presence of the inorganic fertilizers. The bacterial and actinomycete isolates were capable of producing auxins, gibberellins and cytokinins and appeared to use fish emulsion as a source of nutrients and precursors for PGRs. PGR levels in planta following combined treatments of the bacterial and actinomycete isolates and fish emulsion were found to be significantly enhanced over other treatments. The effect of fish emulsion appears to be more related to its role as a nutrient base for the bacterial and actinomycete isolates rather than to the increased activity of the general microflora of treated soil. This is the first report of fish emulsion as a nutrient base for plant growth promoting rhizobacteria. These results also indicate that the successful treatment can be effective and economical for horticultural production in sandy soils such as those found in the United Arab Emirates where fish emulsion is already in use as a substitute or supplement for inorganic fertilizer.     

Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis

The original aim of this work was to increase starch accumulation in potato tubers by enhancing their capacity to metabolise sucrose. We previously reported that specific expression of a yeast invertase in the cytosol of tubers led to a 95% reduction in sucrose content, but that this was accompanied by a larger accumulation of glucose and a reduction in starch. In the present paper we introduced a bacterial glucokinase from Zymomonas mobilis into an invertase-expressing transgenic line, with the intention of bringing the glucose into metabolism. Transgenic lines were obtained with up to threefold more glucokinase activity than in the parent invertase line and which did not accumulate glucose. Unexpectedly, there was a further dramatic reduction in starch content, down to 35% of wild-type levels. Biochemical analysis of growing tuber tissue revealed large increases in the metabolic intermediates of glycolysis, organic acids and amino acids, two- to threefold increases in the maximum catalytic activities of key enzymes in the respiratory pathways, and three- to fivefold increases in carbon dioxide production. These changes occur in the lines expressing invertase, and are accentuated following introduction of the second transgene, glucokinase. We conclude that the expression of invertase in potato tubers leads to an increased flux through the glycolytic pathway at the expense of starch synthesis and that heterologous overexpression of glucokinase enhances this change in partitioning.     

Evaluation of Crocus sativus L. (saffron) on male erectile dysfunction: A pilot study

In this study, the effect of Crocus sativus (saffron) was studied on male erectile dysfunction (ED). Twenty male patients with ED were followed for ten days in which each morning they took a tablet containing 200 mg of saffron. Patients underwent the nocturnal penile tumescence (NPT) test and the international index of erectile function questionnaire (IIEF-15) at the start of the treatment and at the end of the ten days. After the ten days of taking saffron there was a statistically significant improvement in tip rigidity and tip tumescence as well as base rigidity and base tumescence. ILEF-15 total scores were significantly higher in patients after saffron treatment (before treatment 22.15±1.44; after treatment 39.20±1.90, p<0.001). Saffron showed a positive effect on sexual function with increased number and duration of erectile events seen in patients with ED even only after taking it for ten days.     

Growth and accumulation of storage reserves by somatic embryos of Ocotea catharinensis Mez. (Lauraceae)

The focus of this study was the assessment of in vitro growth of embryogenic cultures of Ocotea catharinensis Mez. (Lauraceae) on Woody Plant Medium (WPM) supplemented with 22.7 g l^–1 sorbitol, 2 g l^–1 Phytagel, 20 g l^–1 sucrose and 400 mg l^–1 glutamine and the biochemical analysis of somatic embryos at different developmental stages (globular, early cotyledonary, cotyledonary and mature). The embryoids underwent repetitive embryo-genesis and developed non-synchronously, throughout the culture period. Dry weight increased 12.6- and 26.8-fold after 3 and 5 weeks, respectively. During the 5 week culture period a reduction in the frequency of embryoids at the globular stage and increasing frequencies of embryoids at early cotyledonary, cotyledonary and mature stages were observed. Embryoids at the mature stages presented a small but significantly higher water content than those at the globular stage. Therefore, embryoid expansion at the latter stages of development was mainly due to water uptake. Embryoids at the globular stages accumulated higher levels of total proteins while those at the cotyledonary and mature stage showed higher levels of soluble sugars and starch (physiological markers of embryo maturation). Thus, significant differences in the profiles of accumulated storage reserves were detected in the embryoids at different developmental stages in the culture conditions tested.