Comparative water balance profiles of Orchidaceae seeds for epiphytic and terrestrial taxa endemic to North America

The Orchidaceae have dust-like seeds that use wind currents for long-distance dispersal. Lacking endosperm, orchid embryos consume free-living, mycorrhizal fungi as a carbon source (mycotrophy) after settling on a substrate. Few studies have investigated orchid seed morphology as it relates to ecology, but conceivably variations in seed size and testa characteristics could be linked to water loss rates aimed at maximizing germination in a particular habitat. Seeds of 2 epiphytic, 1 aquatic, and 7 terrestrial orchids native to North America were compared with respect to water balance profiles: Cleistes bifaria, Encyclia tampensis, Epidendrum nocturnum, Habenaria repens, Isotria medeoloides, Liparis elata, L. hawaiensis, Platanthera holochila, P. integrilabia, and P. leucophaea. Water content, water loss rate, activation energy, and equilibrium humidity were assessed for each species. Seeds of epiphytic orchids were smaller, lighter, more porous, and had higher water loss rates compared to terrestrials. No active mechanism for water absorption exists in seeds of either group. Water loss appears to be a species-specific phenomenon that may be linked to the ecological niches these species occupy.     

Somatic Embryos and Plantlets from an Ancient Clone of the Grapevine (cv. Cabernet-Sauvignon) by Apomixis in vitro

Plantlets were produced by induction of nucellar embryony (apomixis)in isolated unfertilized ovules of a non-apomictic plant—thegrapevine (Vitis vinifera L. cv. Cabernet-Sauvignon). Ovulesgrown in liquid culture with benzyladenine (5–10 µM)and ß-naphthoxyacetic acid (5–25 µM) formeda nucellar callus which gave rise to somatic embryos. For growthof plantlets the embryos were transferred to an agar mediumcontaining gibberellic acid (1 µM) and 2-isopentenyladenine(5 µM). This is the first report of somatic embryo formationin a cultivar of a temperate fruit.     

Variation in Two Sorbus Species Endemic to the Isle of Arran, Scotland

A study of the morphology of plants from four species of Sorbus,S. arraensis, S. pseudofennica, S. aucuparia and S. rupicola,two of them endemic to North Arran, suggested that the originof the two endemic groups could have been by hybridization. Variation between individuals within the hybrid groups indicatesthat the two groups, probably of hybrid origin (S. arranensisand S. pseudofennica), could each consist of a set of geneticallyisolated clones reproducing by apomixis, with the possibilityin the case of S. pseudofennica of occasional genetic interchangeby sexual reproduction. The overlap of characters between the two hybrid groups suggeststhat the inflow of genetic material by hybridization and geneticmodification in these groups is continuing. Sorbus arranensis Hedl., Sorbus pseudofennica E. F. Warb., Sorbus aucuparia L., Sorbus rupicola (Syme) Hedl., interspecific hybridization, apomixis     

Apomixis in Coix aquatica Roxb

When plants of Coix aquatica Roxb. were grown in isolation orbagged, with removal of staminate spikelets several producedone or two seeds, and one plant formed several seeds. Thesewere presumably formed through apomixis, of the autogamous type.Apomixis occurs side by side with sexual reproduction, and istherefore facultative. The fact that one of the plants grownunder the same conditions had higher apomictic seed set thanothers, and both its apomictic and selfed progeny also showedhigher apomictic seed formation suggests that these have greaterapomictic potentialities than others. Genetic analysis of apomixissuggests that it is recessive to sexuality, and is probablygoverned by a number of genes. A few triploids tested did notshow any apomictic seed set indicating that polyploidy per semay not be responsible for initiation of apomixis. Except thatit is a diploid, C. aquatica seems to fulfil the criteria forapomixis, yet it reproduces predominantly by sexual means.     

Reproductive Biology of Plantago: Shift from Cross- to Self-pollination

The breeding systems of an obligate outbreeder (Plantago lanceolata)and an obligate inbreeder (P. patagonica) were compared. Outbreedingin the former and inbreeding in the latter species is achievedby self-incompatibility and preanthesis cleistogamy, respectively.The difference in breeding system is accompanied by a differencein the pollen output, pollen to ovule ratio, stigma and anthersize, and seed weight. P. ovata, a third species known onlyin cultivated form, is intermediate. This species has two typesof plants, some of which are protogynous and others where stigmareceptivity and anther dehiscence synchronize. Although thedifferences between the two intraspecific variants are not large,they nevertheless suggest that the species is in evolutionaryflux. Domestication is known to have modified the breeding systemin many crop plants and there is evidence that, in P. ovata,a start in this direction has already been made. Plantago lanceolata, P. ovata, P. patagonica, protogyny, anther, stigma, pollen to ovule ratio     

Apomixis and Polyembryony in the Guggul Plant, Commiphora wightii

The present report is a part of our study on the reproductivebiology of a traditional Indian medicinal plant,Commiphora wightii,a source of guggul steroids. Field examination showed a predominantlylarge number of isolated and groups of female individuals. Onlyone andromonoecious and two exclusively male plants were recorded.Female plants set seed irrespective of the presence or absenceof pollen. Hand-pollination experiments and embryological studieshave confirmed the occurrence of non-pseudogamous apomixis,nucellar polyembryony and autonomous endosperm formation forthe first time in this plant, which is presently threatenedby over-exploitation. Commiphora wightii ; autonomous endosperm; guggul; medicinal plant; non-pseudogamous apomixis; nucellar polyembryony     

外来入侵植物胜红蓟的胚胎学观察及繁殖系统研究

以采自我国广东江门和广州两个种群的胜红蓟(Ageratum conyzoides L.)种子为材料,采用流式细胞种子筛选技术(FCSS)和人工控制授粉实验,对胜红蓟的繁殖系统进行研究,并结合整体透明技术和微分干涉差(DIC)显微镜观察法,对其胚珠发育过程和花药结构进行细胞胚胎学观察。种子筛选结果显示,胜红蓟的种子既可以通过有性生殖产生,又可以通过不需要假受精的无融合生殖产生,属于兼性无融合生殖类型。开放性授粉和套袋处理之间的结实率均较高,分别为88%±1.2%和86.2%±1.2%,两者之间无显著差异;而去雄处理的结实率和开放性授粉、套袋这两种处理之间差异显著。胚珠的细胞胚胎学观察结果发现,胜红蓟的有性生殖胚囊发育方式为蓼型,无融合生殖胚囊的发育方式为山柳菊型。胜红蓟的花粉粒在花药内就开始萌发出花粉管,具有闭花受精特性。研究结果表明闭花受精和兼性无融合生殖等繁殖特性保证了胜红蓟在各种生存环境下的结实量,提高其在新生境中归化和入侵的可能性。     

Apomixis in <Emphasis Type="Italic">Eulaliopsis binata</Emphasis>: characterization of reproductive mode and endosperm development

Apomixis represents an alteration of classical sexual plant reproduction to produce seeds with essentially clonal embryos, stimulating wide interest from biologists and plant breeders for its ability to fix heterosis. Eulaliopsis binata (Poaceae), is identified here as a new apomictic species. Embryological investigation indicates that the developmental pattern of embryo sac formation in E. binata represents gametophytic apospory, the embryo originating from an unreduced cell, without fertilization and the mode of endosperm development was autonomous. Sexual embryo sacs were found with a frequency of 1–4% depending on the biotype. The DNA content of nuclei (C-value) in mature seeds was screened by flow cytometry (FCSS) and demonstrated that the endosperm was derived autonomously without fertilization and the three biotypes of E. binata showed varying degrees of apomixis. The Wide-leaf type showed obligate apomixis whereas the Slender-leaf and the Red-haulm type displayed facultative apomixis. In addition, adventitious embryos were observed on the wall of ovary, integument and nucellus cells, indicating that E. binata produces embryos via a mixture of apospory and adventitious embryony.     

Differences in Patterns of Newly Synthesized Proteins in Imbibing Embryos of After-ripened and Aged Seeds of Agrostemma githago and Comparison of Protein Synthesis In Vivo and In Vitro

Patterns of newly synthesized proteins in imbibing after-ripenedyoung and aged Agrostemma embryos show differences in the dynamicsof cotyledons and axes. In the course of imbibition the terminationof some syntheses in the embryonal parts of aged seeds is delayed.As was shown by in vitro translation, mRNAs coding for the synthesisof some proteins are still present in the embryos when proteintranslation has already finished. Key words: Protein pattern, in vivo protein synthesis, in vitro protein synthesis, after-ripened embryos, aged embryos, Agrostemma githago     

Effects of Gibberellic Acid on Floral Development in vivo and in vitro in the Cleistogamous Species, Lamium amplexicaule L.

Using established developmental markers in the corolla and androeciumof Lamium amplexicaule L., we investigated the apparent inductionof open (chasmogamous — CH) from closed (cleistogamous— CL) flowers after application of GA₃ (0.1 mM). In vivotreatment of potentially CL flower primordia caused cell expansionbut not the increased cell division in anthers and corolla necessaryto convert a CL into a CH flower. Floral primordia that appearedto be of undetermined floral type were grown in vitro on a basalmedium supplemented with kinetin (0.1 p.p.m.) and grew to maturityas CL flowers. On media additionally supplemented with GA₃,flowers underwent anthesis, but no true CH flower was produced.Gibberellins appear to be directly responsible for anthesisin the CH flower; but additional, as yet unknown growth factorsare involved in the switch from CL to CH floral form in a developinginflorescence. floral morphogenesis, Lamium amplexicaule L, henbit, cleistogamy, gibberellin, kinetin, anthesis     

Storage Lipid Accumulation by Zygotic and Somatic Embryos in Culture

In vitro accumulation of storage lipids occurs in zygotic andsomatic embryos of Brassicu napus L. The concentration of sucrosein the medium modified the pattern of storage lipid accumulationin zygotic and somatic embryos. The sucrose concentration atwhich the maximum amount of lipid is accumulated by the twotypes of embryos is different Analysis of fatty acid compositionshowed that the same fatty acids are present in embryos in vivoand those cultured in vitro although there are quantitativedifferences. The possibility of using this type of system forin vitro production of valuable plant metabolites is discussed Embryo cloning, somatic embryogenesis, in cilro culture, storage lipids, Brussica napus, oilseed rape     

Diel cycle in the percentage abundance of parthenogenetic females with embryos of different developmental stages in four species of marine cladocerans

The diel cycle in the percentage abundance of parthenogeneticfemales carrying embryos of different developmental stages inthe marine cladocerans Penilia avirostris, Pseudevadne tergestina,Pleopis polyphymoides and Pleopis schmackeri was studied atthe edge of an artificial rocky shore area in a semi-enclosedbay in Hong Kong. Females carried embryos during both day andnight, but females with fully-developed embryos were found predominantlyat night. The diel cycle in the abundance of females with matureembryos was most pronounced in P. tergestina, and less prominent,but still clearly noticeable in P. avirostris, P. schmackeriand P. polyphymoides. The absence or scarcity of females withmature embryos during daytime could be caused by both selectivepredation by visual predators and nocturnal maturation and releaseof embryos. Juveniles of Acanthopagrus schlegeli (black seabream)were the most abundant planktivorous fish in the study areain spring. Stomach content analyses revealed that these daytimepredators fed intensively on marine cladocerans and exhibiteda strong selection for females with mature embryos. On the otherhand, the gradual decline in the percentage of females withadvanced embryos during the latter part of the night, when feedingby visual predators presumably had not yet begun, suggests thatthere was a tendency for nocturnal release of neonates in marinecladocerans.     

Ontogeny of Somatic Embryos of Vigna aconitifolia, Vigna mungo and Vigna radiata

Callus cultures were initiated from immature cotyledons of Vignaaconitifolia, V. mungo and V. radiata on MS medium supplementedwith NAA, picloram or 2, 4-D. On transfer to L-6 liquid mediumsupplemented with low concentrations of picloram, GA₃ and cytokinins,large number of somatic embryos differentiated from the callus.The cells destined to become somatic embryos divided to formspherical or filamentous proembryos. From the filamentous proembryo,the embryo proper developed either at single or multiple sites.Development of somatic embryos from multiple sites resultedin several embryos connected by a common suspensor at the radicleend. Continued divisions of the proembryos led to globular,heart shaped and dicotyledonary stages of somatic embryogenesis.The somatic embryos of V. mungo and V. aconitifolia differentiatedinto tiny plantlets at low frequency (1%) in liquid suspensioncultures supplemented with zeatin, picloram and GA₃. Vigna aconitifolia Jacq, Marechal, mothbean, Vigna mungo L. Hepper, urdbean, Vigna radiata L. Wilczk, mungbean, somatic embryo     

Ultrastructural Studies of Somatic Embryos of Eucalyptus nitens and Comparisons with Zygotic Embryos Found in Mature Seeds

Although somatic embryogenesis has been observed in tissuesfrom a limited number of Eucalyptus species cultured in vitro,no comparisons have been made of the morphology and structureof eucalypt somatic embryos and zygotic embryos found in matureseeds. We used scanning and transmission electron microscopy,in conjunction with histological analysis, to compare maturezygotic embryos with somatic embryos of the commercially-importanttemperate eucalypt Eucalyptus nitens. Apart from differencesin the nature of the outer coating enclosing both embryo types,somatic embryos of E. nitens were observed to have strong similaritieswith zygotic embryos in seeds in terms of their overall size,morphology and internal cellular organization. Many cells inboth sexually-produced and somatic embryos contained numerouslipid-rich globular bodies. The wider significance of theseobservations is discussed with regard to their potential applicationsin eucalypt plantation biotechnology programmes. Copyright 2000Annals of Botany Company Eucalyptus nitens, shining gum, somatic embryo, tissue culture, ultrastructure, zygotic embryo     

Embryo-culture in Lathyrus

Development of 2-mm embryos of Lathyrus articulatus L. in vitrowas stimulated by placing them initially on an agar medium containing8 or 12 per cent sucrose. Germination took place after transferto a similar medium with 4 per cent sucrose. Successful cultureof 0.5-mm embryos was achieved under similar conditions butit was necessary to supplement the initial medium with 10 percent coconut milk. Using this technique, the hybrid L. clymenumL. x L. articulatus was raised from embryos which would haveotherwise aborted in vivo.     

Cryopreservation of Somatic Embryos of Four Species With and Without Cryoprotectant Pre-treatment

The development of cryostorage procedures for somatic embryosproduced from the tissues of plants that normally propagateby means of desiccation- and (often) chilling-sensitive seeds,and that are unstorable by conventional means, offers a viablealternative to the conservation of this otherwise recalcitrantgermplasm. A cryopreservation procedure utilizing cryoprotectantsand partial dehydration was previously developed for hydratedand germinating Pisum sativum embryonic axes. The present contributionapplies that technology to the somatic embryos of a range ofspecies, viz., Coffea arabica (coffee), Manihot esculenta (cassava),Phoenix dactylifera (date palm) and Pisum sativum (pea) andcompares it with results for material that was partially dehydrated,then very rapidly frozen. Cassava, coffee and date palm showedsimilar recovery from cryopreservation irrespective of the procedure.Pea somatic embryos, on the other hand, recovered best fromcryopreservation when pre-treated with the cryoprotectants,glycerol and sucrose, and then subjected to partial dehydration.Copyright1995, 1999 Academic Press Coffea arabica, Manihot esculenta, Phoenix dactylifera, Pisum sativum, cryopreservation, somatic embryos     

Description of a fertilization-independent obligate apomictic species: Corunastylis apostasioides Fitzg

The Australian midge orchid Corunastylis apostasioides of the tribe Diurideae has completely eliminated any male contribution in the process of seed formation, which occurs directly from the maternal tissue by a process termed apomixis. Here, we report C. apostasioides to be an obligate apomictic species devoid of any sexuality and compare its development to a close sexual relative C. fimbriata (R. Br.) D.L. Jones & M.A. Clem. Apomictic characteristics in C. apostasioides include production of seed in absence of fertilization, frequently closed flowers, production of immature pollen in non-dehiscent anthers, expansion of ovaries despite the lack of fertilization and the absence of a citronella scent that is found in C. fimbriata produced to attract pollinating vinegar flies (Jones 2006). The nature of apomixis in C. apostasioides was examined by ovule histology and amplified fragment length polymorphism (AFLP) in each case drawing comparison with sexual C. fimbriata. In C. apostasioides the central megaspore mother cell undergoes diplosporic apomixis, while additional embryos are derived from nucellar or integument initials formed by sporophytic apomixis. Typical of apomicts, C. apostasioides is polyploid compared to the sexual C. fimbriata. The divergences of C. apostasioides from sexuality to apomictic development are discussed.     

The Culture of Immature Pea Embryos

Pea embryos of a range of developmental stages were culturedin a defined medium in vitro for up to 16 days. The criticalfactor for successful culture was the osmotic pressure of themedium; for the stages studied this was provided by the incorporationof 18 per cent sucrose in the medium. The growth of embryosof a range of genotypes was compared; small seeded genotypescould grow at comparable rates in vitro to those attained invivo. The amount of protein synthesized in vitro was similarto that attained in vivo, whereas slightly higher and lowerlevels of starch and DNA respectively were attained in vitro.The roles of intrinsic and extrinsic factors in regulating embryogrowth were studied by comparing the growth in culture of embryosof different genotypes and of hybrid embryos derived from reciprocalcrosses. embryo culture, pea embryos, hybrid embryos, osmotic pressure     

An Analysis of Seed Development in Pisum sativum: IV. COTYLEDON CELL POPULATIONS IN VIVO AND IN VITRO

A method for quantifying changes in the cell population of Pisumsativum cotyledons during development is described. The methodis based on determining the frequency distribution for cellarea following the random sampling of a single-cell suspensionof cotyledon cells. The population profile of these cells changedprogressively and systematically from a single population, similarin size to meristematic cells, found in embryos less than 3.0mg in fresh weight, to a bimodal population in embryos greaterthan 100 mg fresh weight. This method was used to compare embryosof similar size from two genotypes near-isogenic except forgenes at the r locus. No significant differences were foundbetween the cell population profiles of embryos up to 30 mgfresh weight. However, a significant difference was found betweenembryos with fresh weights of 100 mg, the wrinkled (rr) linehaving a higher mean and maximum cell area (2 951 µm²and 9 240 µm² respectively) than the round (RR) line (2591µm²and 6470 µm²respectively). Comparisons were alsomade between cotyledon cell populations from round (RR) embryosgrown in vivo and in vitro. The most obvious differences werethe higher mean and maximum cell size of the large cell populationof in vitro grown embryos which were twice those found in vivo.Embryos grown to either 30 mg or 100 mg fresh weight in vitrohad a much greater proportion of large cells in the populationwith a corresponding reduction in total cotyledon cell number,compared with similar sized embryos grown in vivo. These data suggest that comparisons between different genotypes,or, between cultured and in vivoembryos, based on morphologicalsimilarities between embryos, may be invalid and subject tomisinterpretation. Key words: Pea, seed development, cell population     

Content and De Novo Synthesis of Cocaine in Embryos and Endosperms from Fruit of Erythroxylum coca Lam

There is controversy as to whether the mature fruit of Erythroxylumcoca var. coca Lam. contains the cocaine alkaloid (benzoylmethylecgonine).In the present study, cocaine was monitored to determine ifit was present in embryos and endosperms of mature fruit ofE. coca var. coca Lam., and if present, the time required forde novo synthesis in imbibing seed. Seeds from mature fruitof E. coca were dissected to separate the embryos from the endosperms.The separated embryos and endosperms were analysed for cocaine.Subsequently, endosperms and embryos from seed imbibed. undera light and dark treatment were separated on days 3, 6, 9, 12and 15 and analysed for cocaine. Cocaine was present in embryos(0.005% of d. wt) and endosperms (0–001% of d. wt) ofmature fruit of E. coca. De novo synthesis of cocaine occurredonly in embryos of seed imbibed under light after day 9 of imbibition. Erythroxylum coca, alkaloid, benzoylmethylecgonine, cocaine, embryo, endosperm, seed imbibition