PARTHENOGENESIS IN TETRAPLOID LILIUM LONGIFLORUM

Emsweller , S. L., and Joseph Uhring . (U.S.D.A., Beltsville, Md.) Parthenogenesis in tetraploid Lilium longiflorum. Amer. Jour. Bot. 49(9): 978–984. Illus. 1962.—Nine maternal polyhaploids from 1 capsule and 1 tetraploid from another were produced following pollination of 2 tetraploid Lilium longiflorum plants with pollen from diploids of the same species. One of the 9 plants had 25 chromosomes; the extra chromosome was identified as a modified D. Two other plants had 2 new chromosomes each and the remaining 6 had 24 unmodified chromosomes. Translocations in meiosis of the tetraploid produced the new chromosomes. One plant obtained from a second capsule had 48 chromosomes. The 9 plants were smaller than diploids and the 48-chromosome plant was considered a diploid until mitosis was observed. The 9 plants originated from unfertilized eggs of the tetraploid, and the 48-chromosome plant presumably from chromosome doubling of an egg cell.     

THE DEVELOPMENT OF THE SEED OF ABUTILON THEOPHRASTI I. OVULE AND EMBRYO

Winter , Dorothy M. (Iowa State U., Ames.) The development of the seed of Abutilon theophrasti. I. Ovule and embryo. Amer. Jour. Bot. 47(1): 8–14. Illus. 1960.—Abutilon theophrasti Medic, is a widespread annual weed which produces an abundance of seed in capsules which mature within 20 days after pollination. Ovule differentiation may be observed at least 8 days before anthesis when a sporogenous cell becomes evident and 2 integuments are initiated. An 8-nucleate embryo sac is produced from the chalazal megaspore approximately 2 days before anthesis. The outer integument of the mature campylotropous ovule consists of 2 cell layers, the inner integument has 6 to 15 cell layers. The initially free-nucleate endosperm becomes cellular betwen 3 and 7 days after pollination. At maturity a thin layer of gelatinous endosperm encases the embryo. The Asterad-type proembryo of Abutilon has a stout suspensor and develops rapidly. Four days after pollination cotyledons are initiated; 4 days later a leaf primordium is evident. Fifteen days after pollination the embryo, which has essentially completed its growth, consists of a large hypocotyl with root promeristem and root cap at its basal end, and 2 flat, folded, leaflike cotyledons enclosing a small epicotyl at its upper end. The epicotyl consists of an embryonic leaf and a stem apex.     

Regeneration of black grama (<Emphasis Type="Italic">Bouteloua eriopoda</Emphasis> torr. torr) plants via somatic embryogenesis

Summary Black grama (Bouteloua eriopoda) is an important forage grass in southwestern USA rangelands. Plants were regenerated by somatic embryogenesis. Surface-disinfested seeds were germinated and the embryonic shoots were excised and cultured on Murashige and Skoog (MS) medium gelled with agar. Callus was induced from apical meristems. Calluses were cultured on MS solid medium with six concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or Dicamba (6-dichloro-o-anisic acid) for 6 wk under light or dark conditions. Somatic embryo induction was greatest on 4.52 μM Dicamba, under light, after transferring to an auxin-free medium. Embryo development progressed from globular torpedo to mature embryos phenotypically identical to those naturally produced in seed. These germinated and grew into intact plants and were established in soil and grown to maturity. To our knowledge, this is the first report of somatic embryo induction and regeneration in black grama grass.     

GRASSLAND COMMUNITY FRACTIONS FROM CENTRAL NORTH AMERICA UNDER SIMULATED CLIMATES

For a broad latitudinal comparison, 6 growth chambers with temperature and light-period controls were programmed to simulate 20 weeks of the growing periods for Devils Lake, North Dakota; Lincoln, Nebraska; Austin, Texas; Corpus Christi, Texas; Mexico City, D.F.; and Flagstaff, Arizona. Clonal populations of Panicum virgatum L., the Andropogon gerardi Vitman–halli Hack. complex, A. scoparius Michx., Sorghastrum nutans (L.) Nash, Bouteloua gracilis (H.B.K.) Lag., and B. curtipendula (Michx.) Torr. composed the experimental community fractions. The short growing periods of the northern and high-altitudinal climates accommodated flowering within 5 populations of the North Dakota and Arizona community fractions. In the other climates, the growing periods, variously incomplete during the 20-week programming, also accommodated flowering of many of the early-maturing populations. Annual endogenous rhythms within the geographic variants were suggested by the different response patterning. The northern clones responded rapidly and showed sensitivity to diverse environmental conditions. Many southern clones, requiring more than 20 weeks for flowering, showed developmental homeostasis under the different climates.     

Establishment of Native Semidesert Grasses into Existing Stands of Eragrostis lehmanniana in Southeastern Arizona

The semidesert grassland in southern Arizona has changed from a native grassland to a scattered Prosopis juliflora var. velutina (mesquite) woodland with an understory of African Eragrostis lehmanniana (Lehmann lovegrass) on many sites. To determine native grass restoration potential, seven species were direct seeded into E. lehmanniana stands that were left alive, burned, sprayed with an herbicide and then either left standing, or mowed. Initial native grass establishment was limited in the live standing treatment but was successful for all other treatments when either June or August sowing was followed by consistent summer precipitation and soil water availability. Four species, Bothriochloa barbinodis (cane beardgrass), Bouteloua curtipendula (sideoats grama), Digitaria californica (Arizona cottontop), and Leptochloa dubia (green spangletop) initially established most successfully, while only Muhlenbergia porteri (bush muhly) had consistently limited or no establishment. E. lehmanniana establishment from the seed bank was increased by canopy removal associated with burning. Densities of native grasses one year after successful initial establishment were much lower than that of E. lehmanniana. A possible revegetation strategy would be to spray emergent E. lehmanniana seedlings and surviving plants with an herbicide during the summer rainy season after spring burning. Native grasses could then be established by sowing in early August of that year or June and August of subsequent years until consistent precipitation produces a native grass stand.     

Observations on the Rate of Early Embryogenesis and Cytomixis in Spring Wheat

The rate of early embryogenesis and cytomixis of spring wheat has been observed. The results obtained are summarized as follows: 2 h. after pollination, the two sperms entered into the egg nucleus and the polar nucleus respectively; after 6 h. triple fusion has been completed, a male nucleolus is discernible in egg nucleus. Twenty-four h. after pollination, 2-celled proembryo can be detectable, after 6 d the differentiation in some embryo has initiated. Three d after pollination, the formation of endosperm cells are proceeding, upto 6 d, the embryo sac are full of endosperm cells. After pollination through full development, the degeneration of the antipodals appears, 4 d later the structure of cell and nucleus disappeared. Only naked nucleoli and chromatin mass are remained. After 8 d the degeneration of antipodals almost has completed. Cytomixis has been seen in the cell of nucellus, endosperms, ovary, proembryo and differentiated embryo and it frequently appeared near the proembryo.     

An improved protocol for carrot haploid and doubled haploid plant production using induced parthenogenesis and ovule excision in vitro

In this work, we describe an improved protocol for induced parthenogenesis and ovule culture of carrot (Daucus carota L.). The effects of pollination with parsley pollen and/or 2,4-dichlorophenoxyacetic acid (2,4-D) treatment on the stimulation of parthenogenesis were studied using heterozygous donor plants of 30 varieties and breeding populations of carrots. Isolated ovules, cultured in vitro, enlarged and developed embryos or calli. The application of 2,4-D on pollinated flowers stimulated callus development but did not increase the frequency of embryo development from ovules and, thus, was not useful for increasing the frequency of haploid plant recovery. The efficiency of embryo development was accession-dependent and varied from 0 to 24.29%. In optimized conditions, most accessions responded by embryo development exclusively. The highest frequency of embryo development was observed from ovules excised from ovaries 20–22 d after pollination with parsley pollen. Among several media used for ovule culture, 1/2-strength Murashige and Skoog medium with 0.06 μM indole-3-acetic acid (IAA) was the best. It allowed the production of embryos at a similar frequency as on the media supplemented with kinetin, gibberellic acid, putrescine, or thidiazuron, but restricted callus development. Most plants obtained were haploids and diploids derived from parthenogenesis, as evidenced by homozygosity at three independent loci based on isozyme and PCR analyses. In total, considering haploids and embryo-derived homozygous diploids together, 72.6% of regenerated plants were of gametic origin.     

Cytological characterization of the embryo-lethal mutantdek-1 of maize

Summary The recessive embryo-lethal mutantdek-1 of maize, showing arrest of embryo development at the proembryo stage, lack of carotenoids and anthocyanins and absence in the endosperm of the aleurone layer, was characterized at a cytological level. Cytofluorimetric analysis excluded endoreduplication or polyploidization events in mutant embryonic cells, in spite of an evident increase in nucleolus and nucleus diameters.The data seem to point to an involvement ofDek-1 in the progression of the embryo toward specific developmental steps and in the differentiation of the aleurone layer in the endosperm. Cellular proliferation is not affected by the mutation, as is shown by DNA replication even after the arrest in development and by the possibility of inducing callus from mutant embryos.Abbreviation DAP days after pollination     

Development of Embryo and Fruit in Ipomoea batatas Lam

This paper deals with the development of the embryo and the formation of the fruit for lpomoea batatas Lam. based on the observation of its flower bud differentiation, megasporogenesis and the development of the female gametophyte, microsporogenesis and the development of the male gametophyte. The pollen grain germinated on the stigma about 10–30 min. after pollination. The pollen tube penetrated the transmitting tissue in the middle of the style between 30–60 min. after pollination. After 2 hours the tip of the pollen tube reached the micropyle. Double fertilization completed after 5 or 12 hours then the zygote and the endosperm nucleus formed. The first mitotic division of the endosperm nucleus takes place about 12 hours after pollination, earlier about 3 hours than the first division of the zygote, the latter gives rise to a terminal cell and a basal cell by a transverse division. The second division is transverse in the terminal cell, forming two cells. The basal cell divides longitudinally into two adjoining cells. The terminal cell becomes the proembryo with four cells, and at the same time, the basal cell becomes the suspensor with four cells after 41–52 hours. The proembryo gradually becomes globular, cordate and torpedo-shaped, respectively about 96–120, 144–156, 168-192 hours after pollination. The cotyledons of the embryo gradually prolongate 10 days after pollination. The embryo almost completes its development within 21–30 days after pollination. he fruit is a capsule. The ovary gradually swells 3–4 days after pollination, then forms fruit, which ripens about 21–30 days after pollination, 2R.=4–8 mm. A fruit contains 1–4 seeds. 7,000 fruits were analysed in 1983, the results are as follows: 64.6% of then with only one seed in a capsule, 31.8% two seeds, 5.48% three seeds and 0.1% four seeds. The seeds are small, 2R. from 3.84 mm to 2.84 mm. The shape and the weight of the seeds are different from each other because of difference in number of seeds within a capsule.     

Direct evidence of pseudogamy in apomictic Brachiaria brizantha (Poaceae)

Brachiaria brizantha is a forage grass that has several apomictic accessions. B. brizantha cv. Marandu is a natural tetraploid aposporous apomict widely cultivated in Brazil. Pseudogamy was detected in this species by observation that seed set is suppressed in plants that have had the stigmas excised from the flowers. The egg cell develops parthenogenetically in the apomictic plants, meaning that fertilisation is necessary for the formation of the endosperm. A thorough knowledge of all the events of seed formation in natural apomictic plants is essential for a complete understanding of this mode of reproduction. In this paper, we show direct evidence of pseudogamy in B. brizantha through the cytological analysis of polar nucleus fertilisation and the determination of triploid level of the endosperm tissue. The development of the male gametophyte gives rise to a reduced tri-celled pollen, the viability of which varies throughout the year, reaching 88% in the peak of the flowering period. Discharge of the male gamete takes place around 10 h after pollination and monospermy is the predominant system observed. Precocious embryony was also observed in these plants; embryos arise from egg cells. Endosperm development followed the free nucleus model and was associated with the presence of an embryo. Cellularisation and reserve uptake occurred 2 days after pollination (DAP) and mature endosperm was observed 8 DAP. The triploid level of the endosperm in the apomictic accession confirmed the 2:1 maternal:paternal ratio of genome contribution in the tissue.     

IMPORTANCE OF TIME OF GERMINATION AND SOIL DEPTH ON GROWTH OF PROSOPIS GLANDULOSA (LEGUMINOSAE) SEEDLINGS IN THE PRESENCE OF A C4 GRASS

In an investigation of the causes of the invasion of woody plants into grasslands, competition between seedlings of Prosopis glandulosa Torr. and Bouteloua curtipendula (Michx.) Torr. was examined. Introduction of P. glandulosa into a B. curtipendula neighborhood significantly reduced P. glandulosa dry mass when compared to P. glandulosa growth alone. The greater the time interval from P. glandulosa germination to addition of B. curtipendula, the less interference the grass had on woody plant growth. Reciprocally, the greater the time interval from B. curtipendula germination to addition of P. glandulosa, the more interference the grass had on woody plant growth. Prosopis glandulosa belowground dry mass was <0.02 g (all in the upper 30 cm of soil) when planted after B. curtipendula at any soil depth, but if planted alone its root dry mass ranged from 2 to 8 g depending on depth. Prosopis glandulosa seedling dry mass increased linearly with soil depth, while B. curtipendula dry mass reached a plateau. In general, belowground dry mass of P. glandulosa was distributed throughout the soil depth examined (decreasing with depth), while 80% of B. curtipendula dry mass was found in the upper 30 cm of soil, suggesting a partitioning of soil resources. Data suggest that P. glandulosa and perhaps other shade-intolerant woody species that establish in grasslands do so in disturbances or vegetation gaps. Gaps may close, but by this time woody plant roots are below grass roots, thus partitioning soil resources and reducing interspecific competition.     

Toward in vitro fertilization in Brachiaria spp.

Brachiaria are forage grasses widely cultivated in tropical areas. In vitro pollination was applied to accessions of Brachiaria spp. by placing pollen of non-dehiscent anthers on a solid medium near isolated ovaries. Viability and in vitro germination were tested in order to establish good conditions for pollen development. Comparing sexual to apomictic plants, apomictic pollen has more abortion after meiosis during the microspore stage and a lower viability and, of both types, only some plants have sufficient germination in a high sugar concentration. Using in vitro pollination with the sexual plant, the pollen tube penetrates into the nucellus and micropyle, but the embryo sac degenerates and collapses. In the apomictic B. decumbens, in vitro pollination leads to the transfer of the sperm nuclei into the egg cell and the central cell. The results are discussed according to normal fertilization and barriers in sexual and apomictic plants.     

NATURE OF THE PLANT COMMUNITY. VI. TEXAS GRASSLAND COMMUNITIES UNDER TRANSPLANTED CONDITIONS

Mc Millan , Calvin . (U. Texas, Austin.) Nature of the plant community. VI. Texas grassland communities under transplanted conditions. Amer. Jour. Bot. 48(9): 778–785. Illus. 1961.—Clones of 7 grass taxa, Bouteloua gracilis (H. B. K.) Lag., B. eriopoda (Torr.) Torr., B. curtipendula (Michx.) Torr., Panicum virgatum L., the Andropogon scoparius Michx. complex, the Andropogon gerardi Vitman-hallii Hack. complex, and Sorghastrum nutans (L.) Nash, were transplanted from throughout their distribution in Texas and studied in an experimental garden at Austin. Restricted to western Texas and Panhandle areas, Bouteloua gracilis and B. eriopoda contained similar early-flowering clones throughout their distribution. Less restricted to western sites, B. curtipendula contained later-flowering types from eastern and central areas. In the 4 remaining, widespread taxa, early-flowering potential characterized clones from western sites. These 4 widespread taxa contain the latest-flowering clones from the coast of southern Texas. Clones of Stipa leucotricha Trin. and Rupr. from a broad area in Texas lacked a flowering gradient. Grassland communities of western Texas and the Panhandle, attuned to short growing seasons and low rainfall, were composed of opportunists, the Bouteloua species, and early-flowering variants within the widespread species. Communities of central Texas in habitats of highly unpredictable moisture pattern and a relatively long growing season contained later-flowering variants. Coastal communities attuned to a long growing season contained the latest-flowering variants.     

Barriers to interspecific hybridization between Vigna unguiculata and Vigna vexillata

Summary Interspecific hybridization between Vigna unguiculata and V. vexillata always failed: no seed was obtained in both crossing directions. Two different barriers to crossability were found: a pre-zygotic barrier and a post-zygotic one. Many abnormalities were observed in pollen-tube development, which reduced the percentage of fertilization to 18–30%. Differences in the percentage of fertilization were detected between the two accessions of V. vexillata involved in the interspecific crosses. The development of the interspecific embryo was analyzed and the embryo and endosperm nuclei always degenerated 5–8 days after pollination. The growth of the embryo stopped at a globular stage, which is too early for excision and in vitro culturing.     

In vitro development of maize immature embryos: a tool for embryogenesis analysis

During monocot embryo development, the zygote goes through a proembryo stage characterized by a radial symmetry and later becomes a true embryo with a bilateral symmetry. In order to determine culture conditions for immature embryonic stages, proembryos and embryos were isolated from controlled pollinated maize plants and developed in vitro. Precise culture conditions were determined for each type of explant: a monolayer system for embryos using NBM medium enriched with maltose (0.25 M) but without hormones, and a bilayer system for proembryo stages using N6 medium supplemented with maltose (0.35 M) and zeatin (3 mM). Morphological, cytological, and in situ hybridization analysis have shown that the resulting embryos (stages 1-2), developed in vitro, were similar to those formed in vivo and subsequently gave rise to fertile plants. This work demonstrates that successful embryo differentiation is dependent on specific parameters including the genotype, the nature of the carbon source, the type and concentration of hormones used and orientation of the embryos on the medium. The potential use of these results for embryo rescue and mutant analysis are discussed.     

BIOSYSTEMATIC STUDIES IN THE BOUTELOUA CURTIPENDULA COMPLEX. V. MEGASPOROGENESIS AND EMBRYO SAC DEVELOPMENT

Megasporogenesis and embryo sac development in the sexually reproducing taxa Bouteloua warnockii (2n = 22), B. media (2n = 20), B. uniflora Vasey var. uniflora (2n = 20), B. uniflora var. coahuilensis Gould and Kapadia (2n = 20), and B. curtipendula var. curlipendula (2n = 40) all were found to be of the Adoxa type, in which all 4 megaspores persist and divide once to form an 8-nucleate embryo sac. On the other hand, evidence indicated that plants of B. curtipendula var. caespitosa with high ancuploid chromosome numbers reproduce by pseudogamous fertilization of an aposporous embryo sac. In this taxon the megaspore mother cell did not go beyond the first anaphase of meiosis and the functional embryo sac developed from a nucellar cell. Although the 8-nucleate embryo sac was typical, a 3-nucleate embryo sac was observed to develop in some cases.     

Decreased DNA content of birch (Betula) chromosomes at high ploidy as determined by cytophotometry

Relative amounts of DNA were determined on telophase nuclei by Feulgen cytophotometry for euploid taxa of birch (Betula) with somatic chromosome numbers of 28, 42, 56, 70, and 84. A direct correlation was found between observed DNA absorbance and chromosome number except for plants of B. papyrifera with 84 somatic chromosomes. The DNA density value for nuclei of the 84-chromosome plants fitted a 12.25 ratio instead of the expected 13.0 ratio. The DNA density value for these plants was calculated to be approximately equivalent to plants which would possess 63 somatic chromosomes. The average DNA value per chromosome was 2.73 for the 84-chromosome plants in contrast to 3.50 per chromosome in each of the lower euploids. Nuclear diameters of the 84-chromosome plants were directly related to chromosome number and not to DNA density value. The genomic number of Betula was considered to be x=7, rather than x=14, since a DNA value equivalent to 63 chromosomes is a multiple of 7 and not 14. Diploid birch species (2n=2x=28), therefore, would actually be tetraploids (2n=4x=28). The reduction in DNA content may be an adaptation for the establishment of higher ploidy in birches.     

The ultrastructure of zygotic embryo development in pearl millet (Pennisetum glaucum; Poaceae)

The ultrastructure, morphology, and histology of zygotic embryogenesis in pearl millet (Pennisetum glaucum) were examined using light and electron microscopic techniques. Embryogenesis was initially characterized by the presence of a vacuolated egg cell and zygote. The increased presence of Golgi bodies in the zygote suggested it was metabolically more active than the egg cell. The first zygotic division resulted in a densely cytoplasmic apical cell and a highly vacuolated basal cell. The club-shaped proembryo displayed a large amount of endoplasmic reticulum (ER) and ribosomes, very few lipids, and a continuous gradient of vacuoles from the highly vacuolated basal suspensor cells to the densely cytoplasmic apical cells. The embryo had well-defined parts by 8 days after pollination, including shoot and root meristems, coleoptile, scutellum, provascular system, and the first leaf primordium. Large increases in ER, lipids, starch, and vacuoles occurred in the scutellum during the maturation of the embryo, except in the provascular cells. Throughout zygotic embryogenesis, embryo cells were connected by plasmodesmata except where intercellular spaces occurred. Ultrastructural, morphological, and histological observations of zygotic embryogenesis in pearl millet are in agreement with previous reports for other grass species.     

On the Fertilization of Oryza sativa L. × Sorghum vulgare Pets.

1.The pollen germination of Sorghum vulgate appeared normal on the stigma of the Oryza sativa, but the pollen tubes grew slowly in the style. Some of the pollen tubes may become enlarged in their tips or sometimes bursting, while others have continued to grow and entered the embryo sacs. 2. The growth rate of the pollen tubes varied widely. A few pollen tubes were observed in the embryo sacs of the materials 2 hours after pollination, but most of them entered the embryo sacs much later. 3. The zygote associated with a paucity of endosperm nuclei was observed in the materials 1 day after pollination. The double fertilization and 8–12-celled proembryo associated with a number of the free nuclei of the endosperm appeared with a rather high frequency (10.3%) in the materials 3 days after pollination. Some of them are normal in appearance and others may show more or less abnormalities. 4. No division figure was found except in one single case in which mitoses have occurred in both the proembryo and the endosperm. It is most likely that in such case the proembryo and the endosperm if left intact might develop further. 5. A 80-celled embryo was the biggest one which appeared in the materials 5 days after pollination. In general, no cells were ever formed in the endosperm, except in one instance among the 7 days materials the endosperm became cellular in micropylar end. In all other cases the endosperm either ceased to develop early or disorganized. The disorganized endosperm materials are considered to be utilized by the embryo. 6. In certain instances the free nuclei of the endosperm were not distributed at random. They were not equal in size and might fuse into giant nuclelei. 7. The most striking feature is that in the embryo sacs, in which double fertilization or proembryo and endosperm have occurred, a dark stained pollen tube was commonly present. This fact leads us to the conviction that in general only if a healthy pollen tube entered the embryo sac, double fertilization can take place and further development can proceed. 8. In certain cases the protoplasm of the embryo cells appeared scanty. It is apparently that the normal metabolism of the embryo was disturbed owing to the lack of nutrient, and the death of the embryo ensued. 9. No differentiated embryo was observed and no mature seeds were produced. The materials fixed 12 days after pollination showed a variety of abnormalities and collapses. The authors believe that the failure of seed production of rice X kaoliang was primarily due to the fact that the pollen tubes in the style grew too slowly to reach the embryo sacs in time. The consequence is that the double fertilization took place only in a late stage while the male and female gametes may have already become unhealthy. In addition, in this late stage the stored starch in the maternal tissues having gradually disappeared, the nutrient supply to the embryo sac was therefore limited and the young embryo and endosperm were finally in starvation.     

Cytogenetics of a new type of barley with 16 chromosomes

In the progeny of a trisomic type for chromosome 6, Purple, a 16-chromosome type was obtained, which had a pair of new metacentric chromosome 6 in excess. The new metacentric chromosome 6 was shorter than any of the 14 chromosomes of normal barley complement and showed a heteropycnotic nature at late prophase in somatic mitosis. At metaphase I in the plants with 14+one metacentric chromosome 6 (2n=15) the chromosome configuration was exclusively 7_II+1_I indicating that the extra metacentric chromosome 6 could not associate with the normal chromosome 6. At diakinesis and metaphase I in the new 16-chromosome plants most of the sporocytes showed 8_IIor 7_II+2_I. Neither tetravalents nor trivalents were observed at meiosis. The chromosome behaviour at anaphase I and later stages of meiosis was regular in general, resulted in a fairly high pollen fertility of about 61 per cent. Seed fertility however, was very low. The transmission rate of the new metacentric chromosome 6 through the pollen was extremely low in 16-chromosome plants. Possible origin of new basic number and B-chromosome in diploid level through trisomic condition was suggested (Summary see p. 138).Contribution No. 141 of the Department of Plant Science, University of Manitoba.