几种具无融合生殖特性的植物多胚和多苗现象的观察

报道6种具无融合生殖特性的植物种子的胚数和萌发实生苗数的观察结果。金桔(Fortunellamargarita (Lour.)Swingle)、蜜桔(Citrus unshiu Marcoritch)和花椒(Zanthoxylum bungeanum Maxim.)具珠心胚,含多胚种子频率分别为97.50％、100％和45.00％;多胚种子的胚数范围分别为2～49,3～54和2～6。草地早熟禾(Poa pratensis L.)、滨草(Elymus rectisetus)和湖北海棠(Malus hupehensis(pampon.)Rehd.)具非减数配子体无融合生殖特性,含多胚种子频率依次为34.25％、8.11％和37.50％;前两种的多胚种子中胚数范围为2～3,后者为2～15。蜜桔、草地早熟禾和湖北海棠种子萌发多苗的频率分别为22.00％、6.14％和2.22％。描述了多胚种子中胚的形态、位置和分布。对6种植物含单胚种子的胚的来源进行了分析。初步结论:1.具无融合生殖现象的植物种子含多胚和萌发多苗的特性可作为寻找具无融合生殖特性的植物的形态学指标;2.对其筛选的材料进行大孢子发生、胚囊形成和早期胚胎发育的研究,以期阐明多胚来源和生殖类型。     

Polyembryony increases embryo and seedling mortality but also enhances seed individual survival in Handroanthus species (Bignoniaceae)

Polyembryony seems to be advantageous to mother plants in detriment of their siblings which face competition since the beginning of seed development. This competition may limit the turnover of embryos into seedlings and their survival ability. We analysed polyembryony frequency and embryo to seedling turnover in three Handroanthus species with sporophytic apomixis. We tested if the embryo number per seed affected seed and embryo morphometry, seedling survival ability and seed individual survival (i.e. survival of at least one seedling per seed). The number of embryos per seed was compared with seedling number at different developmental stages. All 14 populations showed high frequencies of polyembryonic seeds (21–91%). As the number of embryos per seed increased (up to eight embryos/seed), there was a reduction of mean embryo mass, area, seedling length, individual seedling survival ability, and embryo to seedling turnover. There was also an increase in embryo morphological anomalies. However, enhanced seed individual survival was also observed. Thus, the high frequency of polyembryonic seeds and the increase in seed individual survival support the idea that polyembryony represents an alternative reproductive mechanism which can favours these species.     

Sporophytic apomixis in polyploid Anemopaegma species (Bignoniaceae) from central Brazil

Apomixis and polyploidy have been important in the evolution of the angiosperms, and sporophytic apomixis has been associated with polyembryony and polyploidy in tropical floras. We studied the occurrence of polyembryony in populations of tetraploid Anemopaegma acutifolium, A. arvense and A. glaucum from the Brazilian cerrados, and histological features of sexual and apomictic processes were investigated in A. acutifolium. All populations and species were polyembryonic (68.9–98.4% of seeds). Normal double fertilization occurred in most ovules, with exceptions being that 3% of ovules were penetrated but not fertilized and in 4% of ovules both synergids were penetrated. The penetration of both synergids suggests a continuous attraction of pollen tubes and polyspermy. Adventitious embryo precursor cells (AEPs) arose from nucellar and integumental cells of the ovule in pollinated and unpollinated A. acutifolium, indicating sporophytic apomixis. However, further embryo and endosperm development required pollination and fertilization. This pseudogamy also allows concurrent sexual embryo development. Similar polyembryony rates and polyploidy indicated that A. arvense and A. glaucum are also apomictic, forming an agamic complex similar to that observed for some species of confamilial, but not closely related Handroanthus. The co‐occurrence of apomixis and polyploidy in different groups of Bignoniaceae indicates homoplasious origin of these agamic complexes. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 77–91.     

Polyploidy and polyembryony in Anemopaegma (Bignonieae, Bignoniaceae)

Polyploidy is a key process in plant evolution, with the asexual formation of embryos representing a way through which polyploids can escape sterility. The association between polyploidy and polyembryony is known to occur in Bignoniaceae. In this study, we investigate polyembryony in four polyploid species of Anemopaegma: A. acutifolium, A. arvense, A. glaucum and A. scabriusculum as well as in one diploid species, A. album. Polyembryony was observed only in polyploid species. We used seed dissection and germination tests to compare the number of polyembryonic seeds. We tested how the pollen source influences the number of polyembryonic seeds and the number of embryos per seed and tested the correlation between the number of viable seeds per fruit and mean number of embryos per seed. The number of polyembryonic seeds observed by seed dissection was higher than the number of polyembryonic seeds determined by the germination test, with the number of embryos produced per seed being higher than the number of seedlings. The dissection of seeds of A. glaucum indicated that a higher number of polyembryonic seeds and a higher number of embryos were present in seeds from cross-pollination than in seeds from self-pollination. On the other hand, germination tests indicated that a higher number of polyembryonic seeds were present in fruits from self-pollination than from cross-pollination. The mean number of embryos per seed was not influenced by the number of viable seeds per fruit in fruits from open pollination. These results indicate a positive relationship between polyembryony and polyploidy in Anemopaegma.     

Self-fertility and polyembryony in South American yellow trumpet trees (Handroanthus chrysotrichus and H. ochraceus, Bignoniaceae): a histological study of postpollination events

Handroanthus chrysotrichus shows pollination-dependent self-fertility, polyploidy, and adventitious polyembryony, and it is closely related to H. ochraceus, for which apparently conflicting reports of self-incompatibility and apomixis have been published. The present study aims to investigate the polyembryony in these species by means of histological analysis of ovule/seed development in unpollinated, selfed, and crossed pistils/fruits (in H. chrysotrichus only) as well as seed germination experiments. Experimental pollinations were carried out to evaluate breeding systems in the studied populations, and the results indicated self-fertility in both species. Adventitious embryo precursor cells (AEPs) were formed in the ovules of unpollinated, selfed, and crossed pistils. However, unfertilized ovules never develop into seeds, and fertilization/endosperm initiation clearly stimulates the formation of AEPs in pollinated pistils. The inability of AEP-bearing unfertilized ovules to initiate endospermogenesis clearly shows that fertilization is needed for adventitious embryo development. Consequently, formation of AEPs is required but is not sufficient for apomictic reproduction in H. chrysotrichus. Analysis of the positions of multiple embryos in the endosperm indicated that fertilized ovules are able to develop into seeds even in the absence of a zygotic embryo. The development of AEPs in ovules of H. chrysotrichus foregoes the stage in which activation of selfed pistil rejection takes place in H. impetiginosus, a species with late-acting self-incompatibility. Our study supports the hypothesis that the self-fertility in H. chrysotrichus (and perhaps also in H. ochraceus) resulted from the emergence of pseudogamous apomixis, favored by the physiological peculiarities of the late-acting self-incompatibility and possibly related to polyploidy.     

AFLP markers closely linked to a major gene essential for nucellar embryony (apomixis<Emphasis Type="Italic">)</Emphasis> in <Emphasis Type="Italic">Citrus maxima</Emphasis> × <Emphasis Type="Italic">Poncirus trifoliata</Emphasis>

Some citrus varieties express a form of apomixis termed nucellar embryony in which the adventive embryos develop from nucellus tissue surrounding the embryo sac. This trait results in many seeds containing multiple embryos (polyembryony). Inheritance of the frequency of polyembryony was studied in 88 progeny from a cross of Citrus maxima (monoembryonic) × Poncirus trifoliata (polyembryonic). The frequency of polyembryonic seed produced by each progeny was determined by scoring 100–500 seeds for the number of seedlings to emerge from each seed. Two groups of eight individuals from each extreme of the population were chosen for bulked segregant analysis with amplified fragment length polymorphism markers amplified with 256 primer combinations. Candidate markers identified in the bulks as linked to the trait were tested on the 32 individuals used to create the bulks and then on the remaining plants in the population. Five candidate markers tightly linked to polyembryony in P. trifoliata were identified. Specific marker alleles were present in nearly all progeny that produced polyembryonic seed, and alternate alleles were present in nearly all progeny that produced only monoembryonic seed. The region defined by these markers very likely contains a gene that is essential for the production of polyembryonic seeds by apomixis, but also shows segregation distortion. The proportion of polyembryonic seeds varied widely among the hybrid progeny, probably due to other genes. Scoring 119 progeny of a P. trifoliata selfed population for the closely linked markers and the proportion of polyembryonic seeds confirmed close linkage between these markers and polyembryony.     

Reproductive bet‐hedging in a rare yet widespread rainforest tree,Syzygium paniculatum (Myrtaceae)

The rare rainforest tree species, Syzygium paniculatum, is the only known Australian species of the genus to produce seeds that regularly have multiple embryos. Evidence from other species suggests that this is a case of adventitious polyembryony, with the embryos arising from maternal nucellar tissue. In the present study we use microsatellite data to determine whether sexual reproduction does occur and, if it does, to investigate the relative fitness of asexual versus sexual seedlings. Genotyping suggested that the species is a polyploid and our results found very little genetic diversity within and among populations (with a total of nine genotypic combinations across the entire species). The only significant variation was between the three northernmost populations and the other eight populations sampled. Analysis of individual embryos showed that sexually derived embryos did occur in some seeds but that these were not necessarily the fittest. In general, the seedling from the largest embryo is the first to emerge and maintains a competitive advantage over the other seedlings from the same seed. We discuss the ramifications of the low levels of genetic diversity and consider whether there is a direct relationship between polyembryony and the inferred polyploidy of the species. We consider the possible advantages of reproductive bet‐hedging but also highlight the susceptibility of a species with low genetic diversity to extreme stochastic events. Syzygium paniculatum occurs in areas heavily impacted by human activity and these findings should contribute to improved management of this threatened species.     

Polyembryony in Citrus. Accumulation of seed storage proteins in seeds and in embryos cultured in vitro.

Citrus exhibits polyembryonic seed development, an apomictic process in which many maternally derived embryos arise from the nucellus surrounding the developing zygotic embryo. Citrus seed storage proteins were used as markers to compare embryogenesis in developing seeds and somatic embryogenesis in vitro. The salt-soluble, globulin protein fraction (designated citrin) was purified from Citrus sinensis cv Valencia seeds. Citrins separated into two subunits averaging 22 and 33 kD under denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A cDNA clone was isolated representing a citrin gene expressed in seeds when the majority of embryos were at the early globular stage of embryo development. The predicted protein sequence was most related to the globulin seed storage proteins of pumpkin and cotton. Accumulation of 33-kD polypeptides was first detected in polyembryonic Valencia seeds when the majority of embryos were at the globular stage of development. Somatic Citrus embryos cultured in vivo were observed to initiate 33-kD polypeptide accumulation later in embryo development but accumulated these peptides at only 10 to 20% of the level observed in polyembryonic seeds. Therefore, factors within the seed environment must influence the higher quantitative levels of citrin accumulation in nucellar embryos developing in vivo, even though nucellar embryos, like somatic embryos, are not derived from fertilization events.     

APOMIXIS AND POLYEMBRYONY IN PACHIRA OLEAGINEA (BOMBACACEAE)

Baker , H. G. (U. California, Berkeley.) Apomixis and polyembryony in Pachira oleaginea (Bombacaceae) . Amer. Jour. Bot. 47(4): 296—302. Illus. 1960.–Trees identified as Pachira oleaginea Decne. are now widely cultivated in tropical and subtropical Africa. Specimens from Achimota, Ghana, are shown to contain 1—5 embryos in a single seed. The flowers, which open at night, are self-compatible but need to be pollinated for seed-setting to occur. Maturation of the fruit takes about 2 mo. but the seeds germinate soon after shedding. One embryo in each seed is usually much the largest and the only one to become established as a seedling. There is no relation between seed-size and numbers of embryos contained. Histological and cytogical studies show that adventitious embryony is responsible for the excess embryos following the formation of a normal sexually produced embryo (2n=72) and endosperm (3n=108). Adventitious embryos arise from single cells in the nucellus in the vicinity of the micropyle and appear to develop faster than the sexually produced embryo with consequent frequent apomictic reproduction. This is the first report of apomixis in the Malvales, which may have important practical consequences as P. oleaginea is a potential economic plant and may fruit only 3 yr. after sowing, rendering seed-propagation feasible.     

The best of both worlds: Apomixis and sexuality co-occur in species of Microlicia,Melastomataceae

Apomixis, the asexual formation of embryos and seeds, occurs in approximately 18% of angiosperm families. Melastomataceae exhibits a remarkable number of apomictic species, distributed among different tribes. This mode of reproduction has been elucidated in Miconieae, but remains unclarified for other groups, such as Microlicieae. Although apomixis has been previously described for Microlicieae species, the cytological basis for this phenomenon is entirely unknown in this group. Thus, populations of Microlicia fasciculata and M. polystemma were used in order to (a) investigate the presence of autonomous apomixis; (b) verify if this mode of reproduction leads to polyembryony; and (c) investigate whether apomixis may occur in parallel with the sexual process. We tested these species for autonomous fruit set and polyembryony, and pollen viability, and analyzed pollen tube growth. Anatomical techniques were used to elucidate the micro- and megasporogenesis and gametogenesis. The species showed autonomous fruit and seed formation and exhibited polyembryony. Apospory and adventitious embryony were the developmental mechanisms of apomixis in M. fasciculata and M. polystemma, respectively. Both species exhibited low pollen viability. However, some viable pollen, reduced embryo sac formation, natural pollination and pollen tube growth enable sexual reproduction and characterize these species as facultative apomicts. The independence of pollinators for fruit set, uniparental reproduction and the possibility of sexual reproduction, confer reproductive assurance and flexibility, bringing together advantages of sexual and asexual reproduction. In this sense, apomixis may have played an important role in the evolution and diversification of Microlicia, a widely distributed genus in the Brazilian Cerrado.     

Polyembryony in Melastomataceae from Brazilian Cerrado: multiple embryos in a small world

Polyembryony has been commonly associated with apomixis in the angiosperms and seems to be more common than expected, even in biomes where sexual reproduction processes are predominant. Recent studies in Cerrado, the Neotropical savannas of Central Brazil, showed high frequencies of apomixis and polyembryony and indicated these processes as reproductive and evolutionary alternatives for plants in these areas. In this sense, we investigated the occurrence of polyembryony and its relationships with ecological (season and type of dispersal, ploidy, species distribution and breeding system) and taxonomic (tribe) factors in the Melastomataceae, a mostly tropical family already known for its high frequency of apomixis and very common in Cerrado. We collected seeds from 69 populations of 53 species, which were sown in germination chambers. After seed germination, the presence and number of seedlings per seed were evaluated as a method to estimate polyembryony. We encountered 18 species (33.96%) with polyembryony (more than one seedling, or gemellar seedlings, originated per seed) concentrated in species of the tribe Miconieae (64%) and Microlicieae (16.67%), but absent in Melastomeae. Monoembryony was present only in sexual species, while all apomictic species were polyembryonic. In Miconia, the polyembryony was correlated with polyploidy, and monoembryony with diploid species. Polyembryony was more common among species with wide distribution in the Cerrado region, which indicates that the presence of gemellar seedlings is important for establishment and survival of the group in the Cerrado biome.     

咖啡多胚现象与多胚苗形态发育的研究

通过目测筛选出咖啡多胚豆(种子),对其进行离体胚培养获得幼小植株,观察分析了多胚及其幼苗的形态发育特征。结果表明,不同倍性咖啡种质均存在多胚现象,不同材料的双胚率不同(变幅在1.06‰ ~ 4.46‰）,其中四倍体材料普遍比二倍体的高。多胚豆中胚的形态和着生位置各异。多胚再生植株生长发育特点各不相同,一般大胚再生植株生长发育正常;小胚植株长势弱,出现畸形植株及生长过程中夭亡等现象。同一咖啡豆中远离正常胚位的额外胚可能是由胚囊外的体细胞发育而形成的不定胚。     

Polyembryony in non-apomictic citrus genotypes

Background and Aims

Adventitious embryony from nucellar cells is the mechanism leading to apomixis in Citrus sp. However, singular cases of polyembryony have been reported in non-apomictic genotypes as a consequence of 2x × 4x hybridizations and in vitro culture of isolated nucelli. The origin of the plants arising from the aforementioned processes remains unclear.

Methods

The genetic structure (ploidy and allelic constitution with microsatellite markers) of plants obtained from polyembryonic seeds arising from 2x × 4x sexual hybridizations and those regenerated from nucellus culture in vitro was systematically analysed in different non-apomictic citrus genotypes. Histological studies were also conducted to try to identify the initiation process underlying polyembryony.

Key Results

All plants obtained from the same undeveloped seed in 2x × 4x hybridizations resulted from cleavage of the original zygotic embryo. Also, the plants obtained from in vitro nucellus culture were recovered by somatic embryogenesis from cells that shared the same genotype as the zygotic embryos of the same seed.

Conclusions

It appears that in non-apomictic citrus genotypes, proembryos or embryogenic cells are formed by cleavage of the zygotic embryos and that the development of these adventitious embryos, normally hampered, can take place in vivo or in vitro as a result of two different mechanisms that prevent the dominance of the initial zygotic embryo.     

Stomatal size,ploidy and polyembryony in Eriotheca Stellate Trichome Species Complex (Bombacoideae – Malvaceae) in the Cerrados of Brazil

• Geographic parthenogenesis, range expansion of apomictic plants after climate changes, has been described for Northern Hemisphere gametophytic apomicts. But similar trends have been observed for sporophytic apomicts of Cerrado, the savannas in Brazil. Eriotheca pubescens is a common Cerrado tree, an agamic complex of either hexaploid/polyembryonic apomicts or tetraploid/monoembryonic sexual individuals. Some populations have been described as a new species, Eriotheca estevesiae, all included in the Eriotheca Stellate Trichome Species Complex (ESTSC). Since breeding systems and ploidy are clearly associated with polyembryony and stomatal size, we used these ancillary features to map the reproductive and ploidy level traits of E. pubescens and E. estevesiae.
• Leaves and seeds were collected from individuals of 19 populations. Seeds were evaluated for the presence of polyembryony and leaves for stomatal measurements.
• Eight populations were monoembryonic while another eight were polyembryonic and for other three, the embryonic pattern was not readily verified. E. pubescens polyembryonic and hexaploid populations formed a homogeneous group, but monoembryonic plants were more variable. E. estevesiae populations were monoembryonic with smaller stomata. In contrast, some E. pubescens monoembryonic populations further south presented larger stomata. Despite these outliers, possibly mixed populations, stomatal size and embryonic pattern differed from northern to southern populations.
• Embryonic pattern and stomatal size indicated that northernmost populations of Eriotheca STSC (E. estevesiae) are diploid and sexual. Southernmost populations, mostly polyembryonic and with large stomata, are hexaploid and apomictic. This is in agreement with geographic parthenogenesis and range expansion of apomictic lineages to southern habitats available after the last glacial maximum.

     

Sexual,apomictic and mixed populations in <Emphasis Type="Italic">Handroanthus ochraceus</Emphasis> (Bignoniaceae) polyploid complex

Some tropical Bignoniaceae form sporophytic apomictic polyploid complexes are similar to better studied temperate plants. Handroanthus ochraceus is a widely distributed Neotropical savanna tree with both monoembryonic/self-sterile, and polyembryonic/apomictic and self-fertile populations, but lacking chromosome number and morphological comparative studies. We tested if monoembryonic/non-apomictic and polyembryonic/apomictic populations differed in ploidy and morphological features, as a basis to understand evolution and biogeography of these plants. Chromosome number and embryo number per seed were investigated, and uni- and multivariate analyses of flower and leaf morphology were done for five populations of H. ochraceus. We found two pure monoembryonic diploid (2n?=?40), and one pure polyembryonic (62–94% of polyembryonic seeds) tetraploid population (2n?=?80). One of the diploid populations presented only one individual with 3.2% polyembryonic seeds and was considered a non-apomictic population. Another population showed predominantly polyembryonic (27–66% of polyembryonic seeds) tetraploid individuals, but one diploid individual with 2% of polyembryonic seeds, and was considered a mixed apomictic and non-apomictic population. Morphological analyses confirmed breeding system clusters, and that stigma width, as well as pollen grain area, was consistently larger in polyembryonic populations. Polyploid plants showed larger cells, as well as larger organs and other distinctive features, which will be useful to identify apomictic populations and to future taxonomic discussions. The species can be considered an agamic complex with apomixis associated with neopolyploidy. This trend is also found in other tropical sporophytic apomictics, contrasting with the usual reports linking diploidy or paleopolyploidy to this kind of apomictics.     

咖啡多胚现象与多胚苗形态发育的研究

通过目测筛选出咖啡多胚豆(种子),对其进行离体胚培养获得幼小植株,观察分析了多胚及其幼苗的形态发育特征.结果表明,不同倍性咖啡种质均存在多胚现象,不同材料的双胚率不同(变幅在1.06‰～4.46‰),其中四倍体材料普遍比二倍体的高.多胚豆中胚的形态和着生位置各异.多胚再生植株生长发育特点各不相同,一般大胚再生植株生长发育正常;小胚植株长势弱,出现畸形植株及生长过程中夭亡等现象.同一咖啡豆中远离正常胚位的额外胚可能是由胚囊外的体细胞发育而形成的不定胚.     

Survival and performance of the invasive vine Vincetoxicum rossicum (Apocynaceae) from seeds of different embryo number under two light environments

The nonnative vine Vincetoxicum rossicum threatens several ecosystems in the Lower Great Lakes Basin of North America. One feature that may contribute to its invasiveness is the production of some seeds with multiple embryos (polyembryony), which may be beneficial as a bet-hedging strategy in variable environments. However, lower seed reserves per embryo in polyembryonic seeds may entail costs in low-light environments. The effect of seed from three embryonic classes (1, 2, or 3 embryos/seed) on V. rossicum survival and growth was studied under two forest understory light environments: full canopy (shade) or canopy gaps (light) in New York state. Two seedling cohorts were planted, in May 2004 and in May 2005. The survival and growth of seedlings was monitored biweekly for two (2005 cohort) or three (2004 cohort) seasons. For both cohorts, plants grown in canopy shade had reduced survival and growth compared with those grown in gaps. Contrary to expectations, seed embryo number had no effect on the final height, survival, or dry mass of plants in either habitat. Our results suggest that any fitness advantage provided by polyembryony may be habitat (light) dependent and not a general trait that affords V. rossicum a benefit in all habitats colonized.     

RAPD Evidence for Apomixis and Clonal Populations in Eriotheca (Bombacaceae)

Abstract: Two woody species of Eriotheca (Bombacaceae) of the Central Brazilian Cerrados were submitted to RAPD analyses. Both species are bee pollinated and have a similar flower structure, but E. pubescens presents adventitious embryony and apomixis, while E. gracilipes is self-incompatible. The RAPD screening reflects these differences in breeding systems, with very low genetic variation in the apomictic species, while the sexual species presented much higher variability with no similar genotypes among the sampled trees. The results suggest that adventitious embryony in E. pubescens effectively results in clonal populations or population mosaics of clonal individuals. Since recent studies have indicated poly-embryony and possibly apomixis in a number of Cerrado woody species otherwise considered obligatorily allogamous, the RAPD results presented here indicate the technique will be a useful tool to detect clonal populations of apomictic origin among Cerrado woody species with mixed mating systems and will help to assess the importance of apomixis as a breeding system for the Cerrado flora.     

Programmed cell death eliminates all but one embryo in a polyembryonic plant seed

Development of multiple embryos from a single zygote, the phenomenon called monozygotic polyembryony, is a widespread reproductive strategy found in higher plants and especially in gymnosperms. The enigma of plant monozygotic polyembryony is that only one embryo in a polyembryonic seed usually survives while the others are eliminated at an early stage. Here we report that programmed cell death (PCD) is the major mechanism responsible for elimination of subordinate embryos in a polyembryonic seed. Using post-fertilized pine (Pinus sylvestris) ovules, we show that once the dominant embryo is selected and, subsequently, the entire female gametophyte is affected by PCD, the cells of subordinate embryos initiate an autolytic self-destruction program. The progression of embryonic PCD follows a rigid basal-apical pattern, first killing the most basally situated cells, adjacent to the suspensor, and then proceeding towards the apical region until all cells in the embryonal mass are doomed. Our data demonstrate that during polyembryony, PCD serves to halt competition among monozygotic embryos in order to ensure survival of one embryo.     

Assessment of polyembryony in lemon: rescue and in vitro culture of immature embryos

The 27 lemon cultivars analysed could be considered slightly or moderately polyembryonic, with 25 to 43% of seeds being polyembryonic and from 1.3 to 1.6 embryos per seed. On this basis, it is necessary to rescue zygotic embryos at an immature stage. Rescue and in vitro embryo development have been studied in two Citrus limon polyembryonic cultivars. Sucrose (50 and 70 g/l) was combined with Murashige and Skoog and Gamborg’s B5 media and tested for optimal growth response. An important effect of genotype was observed: embryos from cultivar ‘Eureka’ had greater survival, germination percentage, and radical development. While the sucrose concentration in the medium did not have an effect on germination, the medium affected the embryo survival and root development of the seedlings, Gamborg’s B5 medium giving the best results. The ability to form plants in vitro was affected by an increase of embryo developmental stage. The germination and seedling height were greater with embryos of seeds collected 135–150 days after anthesis.