Nuclear DNA amount during sporogenesis and gametogenesis in sexual and aposporous buffelgrass

Nuclear DNA amounts (C values) were measured in Feulgen-stained sections of anthers and ovules of sexual plant B-2s (genotype aaaa) and aposporous cultivar Higgins (genotype AAaa) of buffelgrass (Pennisetum ciliare). The mass of the unreplicated nuclear genome of a gamete equals 1C DNA. In both lines, pollen mother cell nuclei were 4C before leptotene; anther wall, dyad, 1-nucleate pollen, and generative cell nuclei were 2C; microspore tetrad, enlarging microspore, and sperm nuclei were 1C. The tapetum persisted as uninucleate cells with 4C DNA. Archespores (2-4C) of both lines initiated meiosis to form megaspore tetrad nuclei with 1-2C DNA. In B-2s, chalazal megaspores (2-4C) formed reduced 8-nucleate Polygonum type embryo sacs, and sacs at 2- and 4-nucleate stages showed distributions with peaks near C1 and C2, corresponding to G1 and G2 cell cycle phases; this is characteristic of active mitosis. Nuclei of 8-nucleate sacs and of eggs and polars were 1C, indicating chromosomes were not duplicated before fertilization. Antipodal nuclei had levels from 1 to 36C, possibly due to polyteny or endopolyploidy. In Higgins, aposporous initials and 2-nucleate embryo sacs showed bimodal distributions of 2n nuclei with peaks at 2C and 4C DNA. Nuclei of newly formed 4-nucleate Panicum type aposporous sacs and of polars were 2C; aposporous eggs stained too faintly for reliable measurement.Names of products are included for the benefit of the reader and do not imply endorsement or preferential treatment by USDA     

中国鹅掌楸双受精和胚胎发生的细胞形态学观察

本文运用常规手段观察了中国鹅掌楸的双受精和胚胎发生,并探讨了其致濒的原因。中国鹅掌楸的卵细胞极性化不明显,中央细胞的两极核直至受精才与精核同时融合,受精为有丝分裂前型;原始细胞型胚乳,胚胎发生为柳叶菜型。     

Conifer somatic embryogenesis for studies of plant cell biology

Summary Embryogenic cultures have been produced for a wide range of conifers and current methods developed for spruce permit the maturation of high quality embryos that can be desiccated and then germinated to form plantlets. Embryogenic suspensions consisting of immature embryos are an excellent source of regenerable protoplasts. This review considers examples of applications of embryogenic suspension cultures for basic studies in three areas of plant cell biology. a) Immunofluorescence studies of microtubules in mitotic spruce cells reveal focused spindle poles at prophase and anphase, suggesting the presence of microtubule organizing centers (MTOCs). Antibodies known to recognize animal MTOCs do not stain the polar regions but do stain developing kinetochores. b) Embryo-derived protoplasts regenerate directly to somatic embryos. Fluorescence studies of the cytoskeleton in freshly derived protoplasts reveal random cortical microtubules and a fine network of actin filaments. During culture, protoplasts change shape and develop transverse cortical microtubule arrays. Embryonal cells of newly formed embryos possess distinctive arrays of cortical microtubules and networks of fine actin filaments while suspensor cells are characterized by transverse cortical microtubules and longitudinal actin cables. c) Transmission electron microscope studies of endocytosis in spruce protoplasts reveal an endocytotic pathway similar to that described previously for soybean. Uptake results are confirmed using high pressure freeze fixation instead of conventional chemical fixation. Presented in the Session-in-Depth Morphogenesis: Plant Cell and Tissue Differentiation at the 1994 Congress on Cell and Tissue Culture, Research Triangle Park, NC, June 4–7, 1994.     

Seasonal distribution of pollen in the atmosphere of melbourne: an airborne pollen calendar

Environmental monitoring of pollen grains in the atmosphere of Melbourne has been achieved using Burkard volumetric traps. Twenty-two families of flowering plants and confiers were identified in the pollen counts. About 62% of these pollen grains belonged to trees, 20% to grasses and 9% to herbs and weedy plants. During spring and summer, the atmosphere contained about 70% of the total annual pollen count. Tree pollen, predominantly elm and cypress, occurred abundantly in late winter and spring, with grass pollen predominantly in spring and early summer. These three types of pollen grains occurred in significant amounts, together accounting for more than 60% of the total annual catch. A seasonal incidence chart (pollen calendar) for Melbourne based on 2 years observation has been constructed. This pollen calendar is useful in identifying sources of allergies against particular seasonal airborne pollen types. Comparison of the time of occurrence of a particular pollen type using the pollen calendar and the time of allergic symptoms, can lead to accurate diagnosis and preventive measures being taken. This study has confirmed that grass pollen is the major source of allergenic pollen in the external environment triggering hay fever and allergic asthma in spring and early summer in Melbourne, Australia.     

美味猕猴桃和软枣猕猴桃种间杂交花粉管行为和早期胚胎发生的观察

美味猕猴桃（Ａｃｔｉｎｉｄｉａ ｄｅｌｉｃｉｏｓａ Ｎｏ．２６）和软枣猕猴桃（Ａ． ａｒｇｕｔａ）杂交,观察了花粉管在花柱内的行为和早期胚胎发生,结果如下： 花粉粒在柱头的乳突细胞表面萌发,在开放型的Ｖ 形花柱道内生长,生长速度比对照的缓慢, 到达胚珠珠孔处的时间延迟５０ 到６０ ｈ。当花粉管生长到花柱基部时出现波纹状弯曲;顶端膨大或尖细;部分花粉管的直径发生变化;少数花粉管生长极不规则。花粉管中胼胝质沿管壁不规则沉积, 有的不形成胼胝质塞,或整个管壁被胼胝质所覆盖。约有２６．７４％ 的胚珠受精, 并发育成种子,其中正常种子占６８．５０％ ;败育种子占３１．５０％ ,其中约有１１．４１％ 为空瘪种子;未受精胚珠占６１．４５％ 。正常种子及其胚均较对照的小。胚发育为茄型,胚乳为细胞型,合子保持休眠十几天后开始分裂形成胚,传粉后５０ｄ 见到子叶胚     

马尾松成熟合子胚的体细胞胚胎发生和植株再生

采用马尾松（Ｐｉｎｕｓｍ ａｓｓｏｎｉａｎａ Ｌａｍ ｂ．）成熟合子胚为起始外植体,在含２,４－Ｄ １０ ｍ ｇ／Ｌ,ＫＴ和ＢＡ 各４ ｍ ｇ／Ｌ的ＤＣＲ培养基上得到胚发生培养物。将白色半透明的愈伤组织（含早期原胚）在含２,４－Ｄ１．０ ｍ ｇ／Ｌ,ＫＴ和ＢＡ 各０．４ ｍ ｇ／Ｌ的ＤＣＲ培养基上保持并增殖。在附加９０００ ｍ ｇ／Ｌ肌醇的ＤＣＲ高渗培养基上得到粗壮的后期原胚。ＡＢＡ 和活性炭同时使用能促进子叶胚的形成,最高频率为３５．１％ 。在无激素培养基上,成熟体细胞胚萌发并进一步形成完整小植株     

烟草花粉发育过程及不同组织中的内源GUS活性

以 X- gluc作为葡糖苷酸酶 ( GUS)酶反应底物 ,用酶组织化学方法检测了烟草 ( Nicotianatabacum L.)不同组织细胞的内源 GUS。在幼苗的根、茎、叶不同发育时期的花药壁、柱头、胚珠分离的生殖细胞和胚囊中 ,均未检测到内源 GUS活性。不同发育时期的烟草花粉内源GUS活性存在差异 ,在小孢子分裂前后和成熟花粉萌发前后有两个活性高峰。检测花粉内源GUS活性的适宜 p H为 5 .0 ;p H7.0的条件下未检测到内源 GUS。用 2 0 %甲醇或 0 .2mmol/L葡糖二酸内酯处理不能完全抑制内源 GUS活性     

Pollen morphology of Gyrostemonaceae,Bataceae andKoeberlinia

Pollen morphology of Gyrostemonaceae, Bataceae, andKoeberlinia, which have been affiliated with glucosinolate-producing taxa, was examined by field emission scanning and transmission electron microscopy. Pollen grains of Gyrostemonaceae are tricolpate with scabrate-spinulate surface and have a thick, unstratified exine, while those of Bataceae are tricolporoidate with granular surface and have a thin exine with a single, outermost granular layer. Gyrostemonaceae and Bataceae, which had often been considered sister taxa based on palynological similarity and now are considered more distantly related, have a similar spongy ektexine, but differences between them are evident.Koeberlinia, which is recently considered a sister group to Bataceae+Salvadoraceae (with no spongy ektexine), has tricolporoidate pollen composed of a plesiomorphic, stratified exine with columellae. The totality of evidence indicates that, contrary to earlier observations, pollen of Gyrostemonaceae and Bataceae does not closely resemble each other, and that the spongy ektexine, which looks to be similar in TEM sections, is a homoplasy that evolved independently in the two families. Dedicated to the late Prof. Emer. Kankichi Sohma (August 28, 1926–June 26, 1995), who supervised us for our M. Sc. and D. Sc. programs at Tohoku University, Sendai. He died after his 40 years career in palynology; his wide range of interests and enthusiasm for research, and his unfailing encouragement for students are greatly missed.     

Occurrence of aneuploid progenies from an asynaptic amphidiploid ofScilla scilloides (lindley) druce II. Mechanism of production of the various aneuploid progenies

The mechanism of production of the various aneuploid progenies was clarified in the asynaptic amphidiploid plants (2n=34+4f+2F, AABB) ofScilla scilloides. Its asynaptic nature and chromosomal stickiness lead to the unequal segregation at anaphase I (AI) in PMC's. The observed values in 18 segregation patterns, 17:17 to 0: 34, were different from the expected values estimated from random segregation of chromosomes. Nevertheless, the preferential transmission of special chromosomes among genomes A (x=8=a₁−a₈) and B (x=9=b₁−b₉) had not occurred. As the result of unequal segregation, the pollen grains with various chromosome numbers were observed. Almost all of the 200 pollen grains contained chromosome numbers more than 17 (range 8 to 34). The observed values of each chromosome number were roughly similar to the expected values of containing the complete set of genome A or B in the random distribution without preferential segregation of chromosomes at AI. The difference between the index of polien mitosis and the pollen fertility was significant in the Wilcoxon matched-pairs signed rank test and suggested the selection for some genomically unbalanced pollen grains during maturation. Consequently, viable pollen grains with various chromosome constitutions are a few (mean pollen fertility of 5.8%) but might produce many aneuploids by self- and cross-pollination.     

Preparation and fusion properties of protoplasts from mature pollen of Nicotiana tabacum

Summary A method to remove the exine from mature tobacco pollen and to release numerous intact pollen protoplasts has been developed. Post-anthesis binucleate pollen was treated with water, buffered with MES at pH 5.5, for two hours. Rupture of the exine was caused by the force of pollen hydration exposing the intine to subsequent enzymatic maceration. The high osmotic pressure (1000 mOsm·kg^-1 H₂O) of pollen protoplasts required a special maceration medium, 4% KCl (w/v). Action of an enzyme solution containing 1% (w/v) Macerozyme and 1% (w/v) Cellulase gave rise to viable protoplasts within 4 hours. When cultured in a tobacco mesophyll protoplast culture medium, the pollen protoplasts underwent regeneration of a cell wall, formation of various tube-shaped structures, and division of the generative nucleus into two nuclei. Using a PEG/Ca²⁺ method pollen protoplasts were fused with diploid mesophyll protoplasts. Evidence of transfer of chloroplasts into the pollen protoplasts was observed after one day of culture.Abbreviations BCP bromocresol purple - FDA fluoresceindiacetate - MES 2-(N-morpholino) ethanesulfonic acid - PEG polyethyleneglycol