Ultrastructural Observations of Papaver rhoeas Mature Pollen Grains

The mature pollen grain of Papaver rhoeas is bicellular. The vegetative cell contains numerous mitochondria; endoplasmic reticulum is not very extensive and there are few ribosomes and plastids. Golgi bodies are in a very active state. The generative cell is lobed and spindle-shaped. The cytoplasm contains many, generally longitudinally arranged, bundles of microtubules. Other organelles are few in number, and include mitochondria, Golgi bodies and short cisternae of endoplasmic reticulum.     

Ultrastructural and Cytochemical Study on Mature Pollen of Pinus tabulaeformis

The pollen of Pinus tabulaeformis Cart. comprised two prothallial cells, a generative cell and a tube cell which degenerated at pollen maturation. The generative cell had its own cell wall, seperating from the intine of pollen, but with its side wall attached to the infine. Cytoplasmic channels were present on the side of the generative cell wall, which faced to the tube cell cytoplasm. The generative cell differed conspicuously from the tube cell. The main differences include: ( 1 ) The chromatin in the generative cell nucleus was condensed, but was dispersed and had numerous nueleare pores in the tube cell nucleus; (2)There was no microbody in the generative cell but many microbodies were present in the tube cell cytoplasm; (3)More inclusions were present in the tube cell than in the generative cell. Both the generative cell and the tube cells contained lipid bodies and amyloplasts in the cytoplasm, but there were more amyloplasts in the former. The tube cell also contained a few proteins which was absent in the generative cell. In addition, there were numerous mitochondria, polyribosomes, and a few endoplasmic reticulums and dictyosomes in the generative and tube cells. DAPI staining demonstrated numerous cytoplasmic DNA in both generative cell and tube cell. The mode of cytoplasmic inheritance, and the composition, structure and the nature of the pollen wall of P. tabulaefonnis are also discussed in this paper.     

花生成熟花粉的超微结构

花生（Arachis hypogaea L．）成熟花粉为二细胞型,具3个萌发沟,少数有4个。外壁呈蜂窝状。花粉壁由覆盖层、基粒棒、外壁内层及内壁构成。线粒体嵴密集、相互平行,脂体被粗面内质网包围。粗面内质网与外核膜相连,亦与线粒体相连。高尔基体甚少。营养核无核仁及染色质,与生殖细胞相连形成雄性生殖单位（male germ unit）。生殖细胞锤形、有壁,见一末端延伸成长尾状（长8μm）。细胞质含核糖体、线粒体、微管,未见体。在有些生殖细胞核内观察到具双层膜、少量嵴及深色内含物的球形结构,其米来源及本质尚不知,有待进一步研究确定。     

Phosphoproteomics Profiling of Nonsmall Cell Lung Cancer Cells Treated with a Novel Phosphatase Activator

Activation of protein phosphatase 2A (PP2A) is a promising anticancer therapeutic strategy, as this tumor suppressor has the ability to coordinately downregulate multiple pathways involved in the regulation of cellular growth and proliferation. In order to understand the systems‐level perturbations mediated by PP2A activation, we carried out mass spectrometry‐based phosphoproteomic analysis of two KRAS mutated non‐small cell lung cancer (NSCLC) cell lines (A549 and H358) treated with a novel small molecule activator of PP2A (SMAP). Overall, this permitted quantification of differential signaling across over 1600 phosphoproteins and 3000 phosphosites. Kinase activity assessment and pathway enrichment implicate collective downregulation of RAS and cell cycle kinases in the case of both cell lines upon PP2A activation. However, the effects on RAS‐related signaling are attenuated for A549 compared to H358, while the effects on cell cycle‐related kinases are noticeably more prominent in A549. Network‐based analyses and validation experiments confirm these detailed differences in signaling. These studies reveal the power of phosphoproteomics studies, coupled to computational systems biology, to elucidate global patterns of phosphatase activation and understand the variations in response to PP2A activation across genetically similar NSCLC cell lines.     

Nucleic-Acid and Protein Contents of Embryogenic Tobacco Pollen

Anthers of Nicotiana tabacum cv. White Burley containing microsporesin mitosis were cultured for 12-14 d at 23° C in order toinduce mitosis of the vegetative cell (embryogenic grains).After this period the total DNA content of such grains estimatedby gallocyanin-chrome alum photometry was double that of themicrospores in mitosis, whereas the total RNA content was reducedby about one-third. Total protein estimated by naphthol yellowS photometry was approximately the same as at inoculation. Incontrast, total RNA and total protein contents of non-embryogenicgrains in the same anthers were at least four times greaterthan at inoculation. It is suggested that degradation of cytoplasmicinformation concerned in gametophytic differentiation takesplace prior to mitosis of the vegetative cell.     

桃花粉离体萌发和花粉管生长特性研究

采用花粉离体萌发法研究不同培养基组分和培养条件对桃花粉萌发和花粉管生长的影响,同时对不同贮藏温度下的桃花粉寿命进行研究.结果表明:固体培养基与液体培养基对桃的花粉萌发率和花粉管长度影响差异不显著;10%蔗糖是大多数桃品种花粉的最适萌发条件;硼能提高桃花粉的萌发率,但对花粉管的生长没有促进作用;桃花粉在20℃～25℃的培养温度下萌发率最高,花粉管最长;桃花粉萌发率和花粉管长度在培养前3 h内上升最快,3～5 h上升趋势减弱,5 h后基本停止;随着贮藏温度的升高和贮藏时间的延长,花粉生活力呈降低的趋势.     

不同培养条件对黄连木花粉萌发和花粉管生长的影响

以黄连木花粉为试材,采用离体培养法研究了培养基组分和植物生长调节物质对黄连木花粉萌发和花粉管生长的影响.结果表明:花粉萌发和花粉管生长的适宜蔗糖浓度为15%,适宜培养温度为25℃;该培养条件下,花粉萌发率和花粉管长度分别达最大值63.3%和412.1 μm.硼酸、赤霉素(GA3)和吲哚乙酸(IAA)在一定浓度范围内,可以促进黄连木花粉萌发和花粉管生长,浓度过高时起抑制作用;最适宜黄连木花粉萌发和花粉管生长的硼酸浓度、赤霉素(GA3)和吲哚乙酸(IAA)浓度分别为100、50和15 mg/L.     

Proteomic Profiling of the Aleurone Layer of Mature Arabidopsis thaliana Seed

The aleurone layer of mature Arabidopsis thaliana seed plays important roles in seed germination and dormancy. However, the proteomic profile of this cell layer is unknown partly because it is difficult to separate this thin cell layer from the mature seeds. In this study, we have used a simple technique to separate the aleurone layer along with the seed coat following germination of seeds and determined for the first time the putative protein composition of this cell layer. By subjecting the total proteins extracted from the seed coat to 2D gel electrophoresis followed by liquid chromatography/tandem mass spectrometry, we identified four AGI loci, AT4G28520, AT5G44120, AT1G03880, and AT1G03890; all of which belong to the seed storage family of proteins. Because in Arabidopsis the diploid aleurone cells of the seed coat perform protein storage functions similar to that of triploid endosperm of other plant species, it is assumed that the above AGI loci are associated with the aleurone layer of the seed coat.     

A Simple Method for Staining Nuclei of Mature and Germinated Maize Pollen

A sugar acetocannine staining technique has been developed for staining the sperm and vegetative nucleus of mature and germinated maize pollen grains. This procedure is simple, stable and highly repeatable. The physiological properties of the mature maize pollen grains are first adjusted by using an in vitro germinating culture solution. This solution is 15% sucrose and contains 360 ppm calcium chloride dihydrate, and 120 ppm boric acid. One part fresh pollen grains is uniformly mixed with nine parts of the solution and left at room temperature for at least 5 hr. One part of this solution is then mixed with two parts of regular acetocannine stain and left overnight. The color of this mixture is pinkish red or raspberry. The sugar in the mixture helps to increase color contrast between the pollen cytoplasm (light pink) and the nuclei (reddish purple), decreases the frequency of burst pollen, increases pollen expansion, stabilizes pollen figures and automatically seals the coverglass.     

Effects of EDTA and Acidified Medium on the Dedifferentiation of Immature Pollen in a Tobacco Pollen Culture

The addition of ethylenediaminetetraacetic acid (EDTA) to themedium stimulated the dedifferentiation from immature pollengrains to embryogenic cells in the tobacco pollen culture systemdeveloped by Kyo and Harada (1986). The stimulative effect ofEDTA was mainly due to the decrease in the pH of the mediumand was very marked in the pollen at the late-bicellular stage(stage IV), which did not become embryogenic under the originalculture condition. (Received September 5, 1990; Accepted September 27, 1990)     

梅花花粉离体萌发和花粉管生长研究

(南京农业大学园艺学院,南京210095)摘要:研究培养基成分、pH值和培养方式对梅花花粉离体萌发和花粉管生长的影响。结果表明:不同品种梅花花粉离体萌发的最适培养基为ME3+200g.L-1PEG4000(pH5.0),品种‘淡丰后’、‘久观绿萼’、‘喧妍宫粉’和‘月光玉蝶’最高萌发率可分别达到58.6%、60.6%、85.6%和50.7%。PEG4000能显著促进梅花花粉萌发,在培养基各成分中作用最大,不可替代。低浓度(50g.L-1)蔗糖对梅花品种花粉萌发作用不显著,而高浓度(≥100g.L-1)蔗糖明显抑制花粉萌发和花粉管生长。固体和液体培养对梅花花粉离体萌发的影响差异不显著。     

Pollen Lipidomics: Lipid Profiling Exposes a Notable Diversity in 22 Allergenic Pollen and Potential Biomarkers of the Allergic Immune Response

Background/Aim

Pollen grains are the male gametophytes that deliver sperm cells to female gametophytes during sexual reproduction of higher plants. Pollen is a major source of aeroallergens and environmental antigens. The pollen coat harbors a plethora of lipids that are required for pollen hydration, germination, and penetration of the stigma by pollen tubes. In addition to proteins, pollen displays a wide array of lipids that interact with the human immune system. Prior searches for pollen allergens have focused on the identification of intracellular allergenic proteins, but have largely overlooked much of the extracellular pollen matrix, a region where the majority of lipid molecules reside. Lipid antigens have attracted attention for their potent immunoregulatory effects. By being in close proximity to allergenic proteins on the pollen surface when they interact with host cells, lipids could modify the antigenic properties of proteins.

Methodology/Principal Findings

We performed a comparative pollen lipid profiling of 22 commonly allergenic plant species by the use of gas chromatography-mass spectroscopy, followed by detailed data mining and statistical analysis. Three experiments compared pollen lipid profiles. We built a database library of the pollen lipids by matching acquired pollen-lipid mass spectra and retention times with the NIST/EPA/NIH mass-spectral library. We detected, identified, and relatively quantified more than 106 lipid molecular species including fatty acids, n-alkanes, fatty alcohols, and sterols. Pollen-derived lipids stimulation up-regulate cytokines expression of dendritic and natural killer T cells co-culture.

Conclusions/Significance

Here we report on a lipidomic analysis of pollen lipids that can serve as a database for identifying potential lipid antigens and/or novel candidate molecules involved in allergy. The database provides a resource that facilitates studies on the role of lipids in the immunopathogenesis of allergy. Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.     

野生烟草花粉活力与柱头可授性及繁育特性研究

采用TTC法测定2个野生烟草材料(花烟草、哥西氏烟草)和1个栽培品种(K326)的花粉活力及其日变化情况,通过联苯胺-过氧化氢法测定3个烟草材料的柱头可授性、利用直接授粉法测定不同开花天数柱头可授性变化,并通过估算花粉胚珠比(P/O)、杂交指数(OCI)及授粉试验分析3个烟草材料的繁育特性。结果表明:(1)哥西氏烟草的花粉活性(74.9%)显著高于K326(52.2%)和花烟草(45.3%),且K326与花烟草间差异不显著;3个烟草材料花粉活力日变化均呈双峰曲线,峰值分别在13:00与15:00左右,且3个烟草材料的花粉活力日最低值与气温日最高值同时出现在14:00。(2)K326柱头可授性显著高于花烟草和哥西氏烟草,且2个野生烟草间可授性无显著差异;不同烟草的最佳授粉时期不同,哥西氏烟草自开花前1d至花后4d,一直保持较高的柱头可授性;花烟草的最佳授粉时期为花后2~3d;K326的柱头在开花前1d至开花后1d授粉最佳。(3)K326以自交为主,存在异交现象;哥西氏烟草繁育类型为兼性异交,自交亲和;花烟草繁育以异交为主,自交亲和性差。研究认为,野生烟草柱头可授性显著低于栽培品种从而影响其结实性,花烟草结实率低主要是由自交不亲和性造成的,而缺乏有效的传粉机制是造成哥西氏烟草结实性差的主要原因。     

Callus Induction and in vitro Regeneration from Barley Mature Embryos

We have assayed different combinations of nutrient media and growth regulators to induce callus and plant regeneration from explants of root, shoot and leaf, complete seed, and isolated mature embryo of barley (Hordeum vulgare L. cv. Hassan). The best results were obtained with mature embryo in J25-8 medium supplemented with 2.0 mg dm^–3 2,4-dichlorophenoxyacetic acid where about 75 % developed friable calli. Some 80 – 85 % of these calli regenerated barley plants in the same J25-8 medium supplemented with 1.0 mg dm^–3 indole-3-butyric acid and 0.1 mg dm^–3 kinetin.     

应用水稻花粉离体萌发体系观察花粉管内微丝分布

采用非固定、DMSO渗透和异硫氰酸标记的鬼笔环肽（FITC—Ph）染色方法,观察水稻花粉离体萌发过程中花粉管内肌动蛋白微丝的形态和分布。结果表明：（1）水稻花粉水合2min后即可萌发,花粉管生长速度在600～1500μm／h之间。（2）水合而未萌发的花粉粒中,大量较短的梭形微丝束构成微丝网络结构,萌发过程中花粉粒内的梭形微丝束松解,部分微丝转移至萌发的花粉管内沿花粉管纵轴呈束状结构;随着花粉管的伸长,微丝束主要分布在花粉管中前端,但在花粉管顶端区域始终未见明显的微丝束。（3）水合后不能正常萌发的花粉粒内肌动蛋白微丝呈弥散不规则分布,在相同萌发时间生长迟缓的花粉管中,微丝束较少,且主要位于花粉管近萌发孔的部位。表明微丝骨架的形态和分布影响水稻花粉管的萌发和生长。