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.     

Polyembryony and apomixis in Eriotheca pubescens (Malvaceae - Bombacoideae)

Apomixis and adventitious polyembryony have been reported for several species of Bombacoideae, including Eriotheca pubescens, a tree species of the Neotropical savanna (Cerrado) areas in Brazil. However, the origin of polyembryonic seeds and their importance for the reproduction of the species remained to be shown. Here, we analyzed the early embryology of this species to establish the apomictic origin of extranumerary embryos. We also observed the geographic distribution of polyembryony in E. pubescens, and tested if apomixis was related to the source of pollen (self or cross) and population density. Moreover, we tested if polyembryonic apomictic embryos would develop normally into seedlings. In the observed seed primordia, after a relatively long quiescent period, the zygote developed into a sexual embryo concurrently with adventitious apomictic embryos which developed from nucellus cells. Adventitious embryos develop faster than sexual ones and are morphologically similar, so that 44 days after anthesis it was virtually impossible to distinguish and trace the fate of the sexual embryo. Polyembryony is widely distributed in populations some 400 km distant, and only one strictly monoembryonic individual was observed during the study. The number of embryos per seed varied between fruits and individuals but was significantly higher in seeds from cross-pollinations than from selfs, although fruit and seed set after crosses were much lower than after selfs. Embryo development into seedlings depended on their weight at germination, but polyembryonic seeds germinated and produced up to seven seedlings per seed in greenhouse conditions. Adventitious embryony and apomictic seedlings would explain the mostly clonal populations suggested by molecular studies.     

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.     

Cytoembryological Studies on Polyembryonic Line SB-1 of Oryza sativa: Polyembryong and Its Origin

Anatomical observations, genetic and cytoembryological studies were conducted to determine the cytological factors in relation to the production of diploid polyembryony within one embryo sac occurring in polyembryonic line SB-1 of O. sativa L. The results showed that the diploid polyembryony in SB-1 was not originated from the fertilization of synergids, but presumably from the megagametophytes with supernumerary egg cells. Potentially polyembryonic megagametophytes at maturity contained either a conventional 3-celled egg apparatus (92.60% of megagametophytes) or the supernumerary eggs forming either a 4-celled (5.2 %) or 5-celled (2.2 %) egg apparatus. In megagametophyte with one additional supernumerary egg cells were almost situated side by side in adjacent to the synergids and recessed from the micropyle, whereas in megagametophyte with two additional supernumerary eggs, one egg cell occupied the conventional possition and the other two lay above the synergids. The arrangement of embryos in polyembryonic caryopsis at various developmental stages was concordant to that of egg cells in megagametophytes with supernumerary eggs. Karyotype analysis showed that most of the twins and triplets were of 2n--2n and 2n-2n-2n respectively. Polyembryony was observed in Fo and F1 caryopsis of cross SB-1 × Malaihong (Malaihong is of non-polyembryony) and in Fl caryopsis of its reciprocal but not in Fo caryopsis. This implies that polyembryony occurring in SB-1 is of a dominant character of embryo sac. Anatomical observations proved that each seedling from twins or triplets was completely independent with no vascular bundle connected with each other. The causes of polyembryony within one embryo sac, the entry of supernumerary sperms and the origin of supernumerary egg cells are discussed.     

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.     

Allozyme evidence for polyzygotic polyembryony in Siberian stone pine (Pinus sibirica Du Tour)

Approximately 4000 mature seeds from 350 trees in nine populations (12–75 trees per population) of Siberian stone pine were investigated for multiple embryos (polyembryony). Haploid megagametophytes and embryos were genotyped for eight allozyme loci. Eight-yone seeds (2.11%) had more than 1 embryo. Of these, 71 seeds had 2 embryos (1.85%), 6 seeds had 3 embryos (0.16%), 3 seeds had 4 embryos (0.08%) and 1 seed had 6 embryos (0.026%). Allozyme comparison of megagametophytes and embryos could distinquish two types of polyembryony in 56 of the 81 seeds. In 28 seeds (50%) the polyembryony was polyzygotic (independent fertilizations of more than one egg cell in the ovule); 25 seeds (45%) had most likely monozygotic polyembryony (genetically identical embryos resulting from the cleavage of a single proembryo) and 3 seeds had both genetically different and genetically identical embryos. To the best of our knowledge, this is the first genetic evidence for the form of polyembryony in conifer seeds.     

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。     

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.     

Evolutionary constraints on polyembryony in parasitic wasps: a simulation model

Polyembryony involves the production of several genetically identical progeny from a single egg through clonal division. Although polyembryonic development allows highly efficient reproduction, especially in some parasitoid wasps, it is far less common than monoembryony (development of one embryo per egg). To understand what might constrain the evolutionary success of polyembryony in parasitoids, we developed Monte Carlo models that simulate the competition between polyembryonic females and their monoembryonic counterparts. We investigated which simulated life‐history traits of the females allow the monoembryonic mode of development to succeed. Published empirical studies were surveyed to explore whether these traits indeed differ between polyembryonic parasitoids and related monoembryonic species. The simulations predict an advantage to monoembryony in parasitoids whose reproduction is limited by host availability rather than by egg supply, and that parasitize small‐bodied hosts. Comparative data on the parasitoid families Encyrtidae and (to a lesser extent) Braconidae, but not the data from Platygastridae, circumstantially support these predictions. The model also predicts monoembryony to outcompete polyembryony when: 1) hosts vary considerably in quality, 2) polyembryonic development carries high physiological costs, and 3) monoembryonic females make optimal clutch size decisions upon attacking hosts. These multiple constraints may account for the rarity of polyembryony among parasitoid species.     

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 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.     

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.     

Phenomenon of polyembryony. Genetic heterogeneity of seeds

Different concepts of polyembryony and genetic heterogeneity of seeds in flower plants have been reviewed. Different types, ways, and forms of plant reproduction appeared in the course of evolution as a consequences of the attached mode of life and autotrophy. This is ascribed to totipotency, “stemminess” of plant cells, and presence of constantly functioning meristems, which determined to a great extent the system of plant safety. There are two ways of formation of a new individual: sexual process → gamospermy involving meiosis and gamete fusion and asexual process → agamospermy without meiosis and gamete fusion and two types of reproduction: seed and vegetative. Both processes may take place simultaneously in one seed, as a result of which many embryos of different origins are formed: uniparental and biparental inheritance. Traditionally, this phenomenon is called polyembryony. It comprises embryoidogeny (a new category of vegetative reproduction): formation of somatic embryos (= embryoids) in the flower, seed, and on vegetative organs. Genetic heterogeneity is one of the most important characteristics of seeds, which is based on different phenomena, such as embryogeny, embryoidogeny, and gametophytic and sporophytic apomixis. When describing two types of polyembryony, sporophytic (nucellar, integumental, cleavage) and gametophytic (synergidal, antipodal), a great attention is paid to characterization of initial cells of the sexual and adventive embryos. A new concept of apogamety is developed from new positions (totipotency and “stemminess”), which is based on different genesis of cells of the egg and antipodal systems. Five possible pathways of formation of the adventive embryos have been proposed from cells of the egg apparatus. Specific features of the formation of adventive embryos in the case of gametophytic apomixis, such as androgenesis and semigamy, are discussed. Morphogenesis of the sexual and adventive embryos proceeds in the mother organism and is determined by the origin and formation of their initials, types of ovule and embryo sac, and specific features of developmental biology. This determines parallelism in their development. The main difference consists in the way of reproduction: heterophasic and homophasic. The phenomenon of polyembryony and genetic heterogeneity of seeds is essential for development of the theory of reproduction and applied research related to seed productivity of plants.     

Sexual polyembryony in almond

Multiple embryos within the same seedcoat occur spontaneously in certain almond [P. dulcis (Mill.) D.A. Webb] cultivars including 'Nonpareil' and 'Mission'. Seedlings from the same polyembryonic seed are frequently viable, though one of the seedlings often shows weak growth and develops poorly. These dwarf seedlings have previously been reported as haploid. In this work, we have characterized several seedlings from 'Nonpareil' polyembryonic seed, including their germination and later growth. Isozyme and simple sequence repeat markers were used to analyze seedling genetic structure. In addition, individual mitotic karyotypes were determined following root-tip staining. The percentage of twin embryos showing aberrant growth was approximately 30%, with mortality rates of about 90%. The majority of these aberrant seedlings appear to be aneuploids. Most secondary embryos appear to be derived from the primary embryo following normal fertilization.     

Asynchronous expression of duplicate genes in angiosperms may cause apomixis, bispory, tetraspory, and polyembryony

Apomicts that produce unreduced parthenogenetic eggs are generally polyploid and occur in at least 33 of 460 families of angiosperms. Embryo sacs of these apomicts form precociously from ameiotic megaspore mother cells (diplospory) or adjacent somatic cells (apospory). Polysporic species (bisporic and tetrasporic) are sexual and occur in at least 88 families. Their embryo sacs also form precociously, but only non-critical portions of meiosis are affected. It is hypothesized that (i) the partial to complete replacement of meiosis by embryo sac formation in apomictic and polysporic species results from asynchronously-expressed duplicate genes that control female development, (ii) duplicate genes result from polyploidy or paleopolyploidy (diploidized polyploidy with chromatin from multiple genomes), (iii) apomixis results from competition between nearly complete sets of asynchronously-expressed duplicate genes, and (iv) polyspory and polyembryony result from competition between incomplete sets of asynchronously-expressed duplicate genes. Phylogenetic and genomic studies were conducted to evaluate this hypothesis. Apomictic, polysporic, and polyembryonic species tended to occur together in cosmopolitan families in which temporal variation in female development is expected, apomicts were generally polyploid with few chromosomes per genome (X = 9.6pL0.4 SE), and polysporic and polyembryonic species were paleopolyploid with many chromosomes per genome (x= 15.7pL0.6 and 13.2pL0.4, respectively). These findings support the proposed duplicate-gene asynchrony hypothesis and further suggest asexual reproduction in apomicts preserves primary genomes, sexual reproduction in polysporic and polyembryonic polyploids accelerates paleopolyploidization, and pa-leopolyploidization may sometimes eliminate gene duplications required for apomixis while retaining duplications required for polyspory or polyembryony. Hence, apomixis, with its long-term reproductive stability, may occasionally serve as an evolutionary springboard in the evolution of normal and developmentally-novel paleopolyploid sexual species and genera.     

DEVELOPMENTAL PATTERNS IN PINE POLYEMBRYONY

Berlyn , Graeme P. (Yale U., New Haven, Conn.) Developmental patterns in pine polyembryony. Amer. Jour. Bot. 49(4): 327–333. Illus. 1962.—An investigation of multiple-embryo development in pine was undertaken. The occurrence of 4 embryos per mature seed was found in approximately one third of the seeds of Pinus lambertiana and Pinus cembra examined and in a few seeds of Pinus strobus. These embryos were derivatives of the 4 embryo-initial cells of the apical tier of the proembryo. There was a progressive reduction in size and development of embryos in a given seed, the embryo furthest from the micropyle being the largest and most differentiated. However, in many cases, histogenesis proceed even when embryo size was significantly arrested. All 4 embryos in some seeds were viable in vitro, but rarely did more than one embryo germinate in vivo. This may be associated with the more rapid reactivation of the largest embryo. Variation in the extent of multiple-embryo development reflects the apparently indeterminate pattern of competition within an embryo system.     

Embryo and Endosperm Phytochemicals from Polyembryonic Maize Kernels and Their Relationship with Seed Germination

Because of the growing worldwide demand for maize grain, new alternatives have been sought for breeding of this cereal, e.g., development of polyembryonic varieties, which agronomic performance could positively impact the grain yield per unit area, and nutritional quality. The objectives of this study were to (1) determine the phytochemicals present in the embryo and endosperm of grain from maize families with high, low, and null polyembryony frequency, which were planted at different locations, and (2) state the relationship between these compounds and seed germination. The extracted phytochemicals from corn were identified by HPLC-MS. The results showed that the genotype with the highest presence of phytochemicals was the brachytic population with high polyembryony called “BAP”, which also required less water during the germination process. The number of phytochemicals in both embryo and endosperm tissues was not related to the sowing location where they came from or the type of polyembryony. The number of different phytochemicals depended on the grain tissue from where they were extracted. The chemical compounds found in the different maize tissues were related to the development of the plant, either in roots or nibs because these are mainly associated with the lignin synthesis.     

Embryogenesis of Polyembryonic Rice ApⅢ: Structural and Histochemical Studies of Egg Apparatus Around Fertilization

The structural and histochemical changes of the egg apparatus in the polyembryonic rice (Oryza sativa L.), ApⅢ with the highest frequence of additional embryos among the polyembryonic rice investigated, before and after fertilization were studied and compared with those of normal and other polyembryonic rices in a similar developmental period. A total of 2 932 ovules were observed and each of them contained only asingle embryo sac with a set of egg apparatus. Among 1 655 embryo sacs, there were 1 643 embryo sacs (99.27%) with one normal egg apparatus in each embryo sac, and only 12 embryo sacs (0.73%) from the remainder with 4 celled egg apparatus, i.e. two eggs and two synergids. Neither the numerous poly egg apparatus and egg like cells, nor the double set of embryo sacs each containing one egg apparatus and other abnormal egg apparatus in single ovary, which were reported by earlier investigators to have high frequency of embryo production in SB 1 and ApⅣ, were observed. The egg cell was located at the subterminal site of the micropylar end of embryo sac. The cytoplasm of egg cell was rich in protein materials and poly saccharide grains, which did not disappear until the division of zygote. The prominent nucleus was closely surrounded by protein and polysaccharide grains, which did not disappear until the division of zygote. No cytological difference was found between egg cells from the normal and abnormal egg apparatus. The two synergids were fully developed and situated at the upper most part of the micropylar end of the mature embryo sac. In most embryo sacs, the synergids were flask shaped with longer necks, and a widened cap shaped top, in close contact with the micropyle. The synergids had a well developed filiform apparatus. The characteristic appearance of the filiform apparatus as well as the cap neck region of synergids before and after pollen tube penetration were easily distinguishable from the egg cell. The structure, the stainability with Coomassie Brilliant Blue and PAS reaction, the process of accumulation, distribution and disapperance of the cytoplasmic protein materials and polysaccharide grains of the two synergids, the persistent and rarely the receptive synergids before and after pollen tube penetration, were closely similar to those of egg cell of the same developmental stage. In comparison with normal and other polyembryonic rice reported, the size of nucleus and nucleolus and their stainability also strongly resembled those of egg cell. Based on the results observed, the main conclusions are summarized as follows: (1) the additional embryos very frequently developed in the young and mature seed of polyembryonic rice ApⅢ were produced by one or two synergids of normal egg apparatus, rarely by 4 celled egg apparatus; (2) during fertilization, the synergids, in addition to the natural specific function of introducing pollen tube and transferring sperms to egg cell and central cell, could be closely associated with the potentiality to breed one or two additional embryos; and (3) as compared with that of normal or other polyembryonic rice it is firstly disclosed that in a few embryo sacs of ApⅢ, the cytoplasmic and nuclear structure, the active anabolism and catabolism of protein and polysaccharide materials and the delayed disorganization at the mid basal region of the receptive and persistent synergid still remained unchanged before the division of zygote. Such salient features could be the predisposition for the origin of additional embryos in ApⅢ.