Abstract: Three maize genotypes previously shown in the literature to respond to anther culture were tested under various conditions. Studies indicated that embryogenic response ranged from 0 to 100 embryos per 1,000 anthers plated and was significantly lower without cold pretreatment of the anthers. Culture in liquid media tended to produce more embryos than in semi-solid as did the addition of activated charcoal to either liquid or solid culture media. Most results were confounded by plant-to-plant variation which tended to obscure significant differences. In one study, germination rate of androgenetic embryos averaged about 20%, but only 26% of those embryos that germinated completed their reproductive cycle and formed seed albeit through sibpollination since plants could not be selfed. Chromosome counts using root tip squashes indicated that regenerated plants were either haploid or diploid but plants scored as non-diploid yielded as much seed as scored diploids. This suggests that progeny can be recovered even from putative haploids, presumably as a result of “sectoring” in the developing ear. A DNA-specific fluorescent dye was used to visualize the presence of putative embryogenic microspores (PEMs) during the culture period. PEM counts were a function of time in culture and were apparently greater than the number of embryos obtained for a given treatment. The data indicate that, as previously reported for other species, both induction and survival phases also exist in maize anther culture.

Abstract: Recombinant cDNA libraries to poly(A)RNA isolated from mature pollen of Zea mays and Tradescantia paludosa have been constructed. Northern blot analyses indicate that several of the clones are unique to pollen and are not expressed in vegetative tissues. The majority, however, are expressed both in pollen and vegetative tissues. Southern hybridizations show that the pollen specific sequences in corn are present in one or a very few copies in the genome. By using several of the clones as probes, it was found that there are at least two different groups of mRNAs with respect to their synthesis. The mRNAs of the first group represented by the pollen specific clones are synthesized after microspore mitosis and increase in concentration up to maturity. The second group, exemplified by actin mRNA, begins to accumulate soon after meiosis, reaches its maximum by late pollen interphase, and decreases thereafter. Although the actin mRNA and the pollen specific mRNAs studied show very different patterns of initiation of synthesis and accumulation during pollen development, the rates of decline of these mRNAs during the first 60 minutes of germination and pollen tube growth in Tradescantia are similar and reflect the previously observed declines in rates of protein synthesis during this period.

Abstract: Improvement of the yield of green plants was obtained by decreasing the concentration of ammonium nitrate in the culture medium and using glutamine as a nontoxic nitrogen source. Embryoids developed directly from the microspores on such media, and frequently green plants were obtained by germination of the embryoids on the induction medium, whereas formation of calli was greatly reduced.

Abstract: Mechanical isolation of Brassica napus microspores from whole buds with a micro-blender enabled the rapid isolation of large numbers of microspores free from tissue and cellular debris The procedure resulted in savings of time and labor and could be used independent of the size and number of buds to be examined Between 700 and 1000 embryos per bud were obtained Compared to anther removal there was no difference in embryo yields or quality Microspore isolation under cool conditions and overnight incubation prior to plating improved the frequency of embryogenesis Over 75% of the embryos developed into normal torpedo-stage structures if the medium was replenished during culture and the embryos placed on a gyratory shaker Over 80% of the torpedo-shaped embryos would ultimately develop into plants The implications of these techniques to genetic and physiological studies are discussed

Abstract: SUMMARYSpontaneous intercellular chromatin migration (cytomixis) occurred in PMCs of Lilium davidii at synizesis stage of meiosis, and less frequent in later stages. This process has led to the formation of up to 9.44% to 13.6% of PMCs, microspores, and generative cells with chromosome numbers deviating from the normal haploid chromosome complement, n= 12. The abnormal cells contained chromosome numbers ranging from 8 to 15. The patterns of intercellular chromatin migration may be divided into two types: (a) The chromatin substance may migrate from one nucleus into one adjacent cell, and (b) from one nucleus into two or more cells or from two or more nuclei into one cell. According to the chi-square test, it has been shown that the differences between the patterns of intercellular chromatin migration and the variation of chromosome number in the PMCs, microspores or generative cells are not significant. Because of cytomixis, about 10% cells proved to be aneuploids, with hypoploids predominating. It shows ...

Abstract: Maize anthers have been induced on modified N6 medium to produce embryoids. Different stages from the cultures were sampled and prepared for microscopical examination. The microspores at the onset of culture were in an early developmental stage, with the nucleus and numerous organelles centred in the middle, surrounded by many small vacuoles with a lipid content. The binuclear pollen grains contained small vesicles and much starch. The partially condensed vegetative nucleus indicated participation of the vegetative component in the formation of multicellular pollen grains (MPGs). Several MPGs have been observed which differed in morphology. We suggest, on the basis of these ultrastructural observations, that in maize mainly the vegetative cell contributes to the MPG which further develops directly into embryoids.

Abstract: The histochemistry of different developmental stages of the pollen wall, aperture, and Ubisch bodies of Triticum aestivum is examined with light and transmission electron microscopy. Various parts of the callosic envelope of the tetrad spores stain differentially. At the late tetrad stage, the probacules and the coat of pro-Ubisch bodies are densely stained for acidic polysaccharides, protein, and neutral polysaccharides. The protectum and the core of pro-Ubisch bodies are moderately stained. Upon release of microspores from the callosic cell envelope, the stainability for acidic polysaccharides increases in the exine and in the wall of Ubisch bodies, becoming very intense in the wall of mature pollen grains and Ubisch bodies. The stainability for neutral polysaccharides is decreased in the mature pollen wall and in the Ubisch bodies, while the stainability for protein increases. The results also indicate the probability of the presence of unsaturated lipids and the absence of free aldehydes in the pollen wall and Ubisch bodies.

Abstract: Anthers of wheat cultivars Orofen and Pitic 62 were incubated for 8 days at 15, 20, 25, 30, 35 and 40°C before transfer to 25°C. Compared with anthers cultured at 25°C constantly, anthers treated at 30°C produced 40% more microspore callus and green plants in both cultivars whereas those treated at 35°C produced 2–3 fold more green plants. Treatment at 40°C was deleterious. Possible modes of action of high temperature on callus production and albinism were discussed.

Abstract: Microsporogenesis was investigated in hermaphroditic and male-sterile plants in nine gynodioecious taxa of Hawaiian Bidens. Normal microsporogenesis in hermaphrodites and the onset of abortion in male steriles were similar in all taxa and in a hybrid between two gynodioecious species. The early abnormal vacuolation of tapetal cells is the first visible evidence leading to premeiotic abortion of microsporogenesis in male steriles. The sporogenous cells disintegrate rapidly after the vacuolation of the tapetum, resulting in a shrunken, indehiscent anther which is composed of only the epidermal layer with some remnant cells of the endothecium and the connective at anthesis. In hermaphrodites, the tapetal cells remain dense and undergo karyokinesis to become binucleate during meiosis I. The tapetum becomes plasmodial after microspores are released from tetrads and gradually disappears during pollen formation. The genetic factor(s) which cause the abortion act with remarkable precision and consistency in all taxa investigated. This suggests that gynodioecy in all Hawaiian Bidens is homologous and the establishment of male sterility in Hawaiian Bidens occurred only once. The spread of the genetic male-sterile factor(s) may be the result of adaptive radiation of the original gynodioecious species or natural interspecific hybridization.

Abstract: Crosses were made between members of the two West African okra types ‘Soudanien’ and ‘Guineen’. All crosses succeeded in both directions and the F1 plants which showed hybrid vigour for plant stature were partially sterile. Cytological observations of the F1 plants revealed abnormal meiosis which resulted in the production of microspores of variable sizes. The frequency of viable pollen (as indicated by acetocarmine staining) was low in the hybrids: 35.80% (U.I.92× U.I.313) and 39.41% (1bk-1×U.I.215). The number of seeds produced per fruit was low in the hybrids and only a few of these seeds are viable. The possibility of gene transfer between the two okra types was discussed.

Abstract: Fusion of the generative and vegetative nuclei physically separated by a wall has been observed in cultured microspores of barley. The generative cell appears to play an active role in fusion as it elongates toward the vegetative nucleus, becomes detached from the microspore wall, and finally completely encloses the vegetative nucleus. The generative cell wall disappears before nuclear fusion takes place. Since these events have been known to occur during pollen development in vivo, it is hypothesized that the occurrence of nuclear fusion in cultured microspores is the result of continued expression of the genes for gametophytic development.

Abstract: Successful isolation of haploids (2n=2x=24) from 4x(2EBN) Mexican species was achieved by using Modified Wenzel (MW) and Nitsch and Nitsch (NN) media for anther culture. Both media, MW and NN, gave consistent regeneration of plantlets from 4x(2EBN) Mexican species, and responses to the media were both species and accession (PI) specific. Plantlets regenerated directly from microspores, by-passing the callus cycle, were mostly haploid (2n=2x=24). Haploid plants appeared smaller and weaker than their parents with a drastic reduction in both male and female fertility. Abnormalities observed during meiosis and the lack of success in crossing confirmed the effect of the meiotic irregularities observed in the haploids. A total of 290 seeds was obtained from the 4x × 2x crosses of 4x Mexican species with irradiated pollen of 2xS. cardiophyllum and 2xS. chancayense; however, no haploid (2n=2x=24) plants resulted. Extraction of haploids from colchicinedoubled 4x (2EBN) or natural 6x(4EBN) Mexican species using a Phureja pollinator was examined to provide clues to the potential of using 2x (1EBN) pollinators for haploid induction in 4x(2EBN) × 2x(1EBN) crosses.

Abstract: This invention relates to a process for fusing protoplasts derived from haploid Brassica tissue, comprising the steps of (A) obtaining haploid microspores from a plant grown from diploid Brassica seed; (B) culturing said microspores to produce a culture; (C) deriving from said culture a plurality of haploid protoplasts capable of fusion to form a regenerable diploid synkaryon; and (D) inducing fusion among said plurality of haploid protoplasts to produce a regenerable diploid synkaryon, to regenerable diploid synkaryons produced by the process of the invention, and to plants and seeds derived therefrom.

Abstract: Light and fluorescence microscopy were used to study coenocytic microspore germination from male-sterile (ms1 ms1) soybean plants. Anther squashes from male-sterile plants revealed that a low frequency of natural coenocytic microspore germination occurred in male-sterile anthers of four independent lines; [ms1-North carolina (T260H),ms1-Urbana (T266H),ms1-Tonica (T267H), andms1-Ames (T268H)]. Abnormalities such as giant tubes, branched tubes, tubes with swollen areas, and multiple tubes were observed from coenocytic microspores from all four lines. The Urbana line, however, demonstrated a higher percentage of coenocytic microspore germination than did the other three lines. Flowers of the Urbana line from both malefertile and male-sterile plants, as well as gynoecia pollinated with coenocytic microspores from sterile plants, were used for in vivo studies. Pollen-tube growth appeared normal in male-fertile plants. In contrast, coenocytic microspore tubes rarely were observed in gynoecia from male-sterile plants or in gynoecia from malefertile plants that had been artificially cross-pollinated withms1 ms1 plants. Few tubes from coenocytic microspores were observed in the vicinity of the micropylar region. A low frequency of seed set was achieved in the greenhouse on Urbana male-sterile plants grown in the absence of male-fertile plants. Thus, we believe either that some gametes from coenocytic microspores are able to participate in fertilization at low frequency or that apomixis occurs inms1 ms1 plants.

Abstract: Viruses and cyst nematodes cause many diseases in potato, and therefore, one of the major breeding aims in potato research is to increase the level of resistance. Success in this field may be obtained efficiently by the combined application of classical and cell culture approaches. For Solarium tuberosum, an analytical synthetic breeding scheme has been proposed using the different ploidy levels (Wenzel et al. 1979). This paper concentrates on experiments for a rapid incorporation and acceleration of resistance into breeding clones using the lx, 2x and 4x ploidy levels, which are either obtained partheno- or androgenetically. Microspore culture within the anther as well as in isolation from the anther is used, furthermore, procedures are described to increase genetically the frequency of the in vitro response, particularly the frequency of the regeneration from microspores.

Abstract: In 3 populations ofApluda mutica, a useful forage grass, the anther wall is 4-layered and its development corresponds to the Monocotyledonous type. Successive cytokinesis in microspore mother cells results in isobilateral tetrads of microspores. Pollen is shed at 3-celled stage. The female gametophyte is of the monosporic Polygonum type. Megaspore tetrads and female gametophyte degenerate at different stages of development and this results in poor seed set. The ovule is bitegmic, pseudocrassinucellate and hemianatropous. The integuments and the nucellus get consumed after fertilization. The pericarp forms a thin-layer around the endosperm and mature embryo and the latter is of the panicoid type.

Abstract: The ovule and anther development of Helichrysum rupestre var. errerae from Pantelleria is studied. The ovule is anatropous, tenuinucellate, unitegmic, endotheliate and is derived from a two-zonate primordium. The archespore is one- or two-celled and all the megaspores show a functional tendency, although only the chalazal one is able to form the gametophyte according to the Polygonum type. The polar nuclei fuse before fertilization and the three antipodal cells divide further. Anthers are tetrasporangiate and their wall development conforms to the Dicotyledonous type. The epidermis persists at maturity and the endothecial cells thicken their walls. The tapetum is of the periplasmodial type and its cells become plurinucleate during microsporogenesis. The number of microsporocytes varies along the length of the same locule. Arrangement of the resulting microspores is tetrahedral, isobilateral or decussate. Pollen grains are three-celled when shed. Degenerating microsporocytes and microspores have f...