Resolution of Channels in the Exine by Translocation of Colloidal Iron

Author(s):  
John R. Rowley

The morphology of the exine of many pollen grains, at the time of flowering, is such that one can suppose that transport of substances through the exine occurred during pollen development. Holes or channels, microscopic to submicroscopic, are described for a large number of grains. An inner part of the exine of Epilobium angustifolium L. and E. montanum L., which may be referred to as the endexine, has irregularly shaped channels early in pollen development although by microspore mitosis there is no indication of such channeling in chemically fixed material. The nucleus in microspores used in the experiment reported here was in prophase of microspore mitosis and the endexine, while lamellated in untreated grains, did not contain irregularly shaped channels. Untreated material from the same part of the inflorescence as iron treated stamens was examined following fixation with 0.1M glutaraldehyde in cacodylate-HCl buffer at pH 6.9 (315 milliosmoles) for 24 hrs, 4% formaldehyde in phosphate buffer at pH 7.2 (1,300 milliosmoles) for 12 hrs, 1% glutaraldehyde mixed with 0.1% osmium tetroxide for 20 min, osmium tetroxide in deionized water for 2 hrs and 1% glutaraldehyde mixed with 4% formaldehyde in 0.1M cacodylate-HCl buffer at pH 6.9 for two hrs.

Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.


1979 ◽  
Vol 57 (18) ◽  
pp. 1946-1950 ◽  
Author(s):  
S. C. Agarwala ◽  
C. Chatterjee ◽  
P. N. Sharma ◽  
C. P. Sharma ◽  
N. Nautiyal

A reduction in the size of tassels, male flowers, and anthers resulted from molybdenum deficiency in maize. In molybdenum-deficient plants, anthesis was suppressed or delayed and the anthers had fewer and smaller pollen grains that lacked dense cytoplasmic contents, appeared shrivelled, and had poor viability. Because of molybdenum deficiency, there was a decrease in the activity of starch phosphorylase in mature and freshly shed pollen grains and decreases in the activities of invertase and acid phosphatase at all the five stages at which pollen grains were assayed for the enzymes. The activities of catalase and peroxidase were increased by molybdenum deficiency at all five stages and that of ribonuclease at four out of the five stages of pollen development.


2009 ◽  
Vol 2009 ◽  
pp. 1-12 ◽  
Author(s):  
K. Zienkiewicz ◽  
E. Bednarska

The aim of the present work was the characterization of nuclear bodies in the microspore and developing pollen cells ofHyacinthus orientalisL.. The combination of Ag-NOR, immunofluorescence and immunogold techniques was used in this study. The obtained results showed the presence of highly agyrophylic extranucleolar bodies in microspore and developing pollen cells, which were finally identified as Cajal bodies. In all cases, a strong accumulation of snRNP-indicating molecules including TMG cap, Sm proteins and U2 snRNA, was observed in the examined nuclear bodies. In contrast to their number the size of the identified structures did not change significantly during pollen development. In the microspore and the vegetative cell of pollen grains CBs were more numerous than in the generative cell. At later stages of pollen development, a drastic decrease in CB number was observed and, just before anthesis, a complete lack of these structures was indicated in both pollen nuclei. On the basis of these results, as well as our previous studies, we postulate a strong relationship between Cajal body numbers and the levels of RNA synthesis and splicing machinery elements in microspore and developing pollen cells.


1994 ◽  
Vol 74 (2) ◽  
pp. 357-358 ◽  
Author(s):  
J. H. M. Willison ◽  
C. H. Cheung ◽  
M. I. N. Zhang ◽  
X. Xiao

Turnip (Brassica rapa L.) root tissue was exposed to freeze-thaw stresses of −7, −9, -−11 and −19 °C. The post-thawed tissues were either subjected to leaching in deionized water for 2 h or left at 100% humidity. Tissue survival was then assayed by vital staining using modified 2,3,5-triphenyltetrazolium chloride (TTC) staining in 0.2 M phosphate buffer. Tissue survival was significantly lower for leached samples than for non-leached samples. It is concluded that freeze-thaw injury in plant tissues is enhanced by post-thaw leaching in water. The 0.05 M phosphate buffer commonly used for TTC staining also damaged freeze-thaw injured tissue. Key words:Brassica rapa L., 2,3,5-triphenyltetrazolium chloride (TTC), freeze-thaw injury, leaching


Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1287
Author(s):  
Liai Xu ◽  
Tingting Liu ◽  
Xingpeng Xiong ◽  
Weimiao Liu ◽  
Youjian Yu ◽  
...  

The pollen grains produced by flowering plants are vital for sexual reproduction. Previous studies have shown that two CCCH-type zinc-finger protein genes in Brassica campestris, BcMF30a and BcMF30c, are involved in pollen development. Due to their possible functional redundancy, gain-of-function analysis is helpful to reveal their respective biological functions. Here, we found that the phenotypes of BcMF30a and BcMF30c overexpression transgenic plants driven by their native promoters were similar, suggesting their functional redundancy. The results showed that the vegetative growth was not affected in both transgenic plants, but male fertility was reduced. Further analysis found that the abortion of transgenic pollen was caused by the degradation of pollen contents from the late uninucleate microspore stage. Subcellular localization analysis demonstrated that BcMF30a and BcMF30c could localize in cytoplasmic foci. Combined with the studies of other CCCH-type genes, we speculated that the overexpression of these genes can induce the continuous assembly of abnormal cytoplasmic foci, thus resulting in defective plant growth and development, which, in this study, led to pollen abortion. Both the overexpression and knockout of BcMF30a and BcMF30c lead to abnormal pollen development, indicating that the appropriate expression levels of these two genes are critical for the maintenance of normal pollen development.


2020 ◽  
Vol 11 ◽  
Author(s):  
Hyun Min Kim ◽  
Se Hee Park ◽  
Sang Hoon Ma ◽  
Seo Young Park ◽  
Chul-Ho Yun ◽  
...  

Abscisic acid (ABA) is a key signaling molecule that mediates plant response to stress. Increasing evidence indicates that ABA also regulates many aspects of plant development, such as seed germination, leaf development, and ripening. ABA metabolism, including ABA biosynthesis and degradation, is an essential aspect of ABA response in plants. In this study, we identified four cytochrome P450 genes (CaCYP707A1, 2, 3, and 4) that mediate ABA hydroxylation, which is required for ABA degradation in Capsicum annuum. We observed that CaCYP707A-mediated ABA hydroxylation promotes ABA degradation, leading to low levels of ABA and a dehydration phenotype in 35S:CaCYP707A plants. Importantly, seed formation was strongly inhibited in 35S:CaCYP707A plants, and a cross-pollination test suggested that the defect in seed formation is caused by improper pollen development. Phenotypic analysis showed that pollen maturation is suppressed in 35S:CaCYP707A1 plants. Consequently, most 35S:CaCYP707A1 pollen grains degenerated, unlike non-transgenic (NT) pollen, which developed into mature pollen grains. Together our results indicate that CaCYP707A mediates ABA hydroxylation and thereby influences pollen development, helping to elucidate the mechanism underlying ABA-regulated pollen development.


2006 ◽  
Vol 33 (6) ◽  
pp. 547 ◽  
Author(s):  
Reinhard K. Proels ◽  
Mari-Cruz González ◽  
Thomas Roitsch

The tomato extracellular invertase family comprises four members with different expression patterns. Among the three invertase isoenzymes expressed in floral tissues, Lin5, Lin6 and Lin7, the expression of Lin7 was previously shown to be restricted to the tapetum and pollen. Histochemical analysis of β-glucuronidase (GUS) reporter activity shows Lin7 expression in pollen and pollen tubes of corresponding transgenic plants. The physiological relevance of the identification of gibberellin-responsive cis-acting elements for induction of the Lin7 promoter is supported by the repression of Lin7 expression in pollen grains by the gibberellin biosynthesis inhibitor paclobutrazol. Functional approaches with transgenic tomato plants establish a link between gibberellin action and invertase function in the tapetum for pollen development: both tissue-specific antisense repression of extracellular Lin7 and ectopic inactivation of the biologically active GAs by expression of a GA2-oxidase under control of the Lin7 promoter result in germination deficient pollen. These complementary findings support the idea that the GA requirement of pollen development, pollen germination and pollen tube growth are linked to energy metabolism via the regulation of an extracellular invertase as a key enzyme for carbohydrate supply via an apoplasmic pathway.


2001 ◽  
Vol 7 (S2) ◽  
pp. 452-453
Author(s):  
Thomas H. Johnson ◽  
Steven J. Schmitt ◽  
Karen S. Renzaglia

Among land plants, lower vascular plants produce the most elaborate of all male gametes. in order to characterize the degree of variability in sperm cell architecture among ferns, this scanning electron microscope study was undertaken on three highly disparate genera: Ceratopteris(a derived leptosporangiate fern) Angiopteris(a derived eusporangiate fern) and Psilotum(a putatively basal eusporangiate fern).Mature gametophytes were placed in 0.01M phosphate buffer (pH ca. 7.2) or distilled water until spermatozoids were released. Gametophyte portions were removed and the spermatozoid suspension fixed with 2% glutaraldehyde in 0.05M phosphate buffer. Post-fixation utilized 1% aqueous osmium tetroxide for lhr, followed by rinsing and dehydration in ethanol (10 min @ 25, 50, 75 and 100%). The dehydrated pellet was suspended in 100% hexamethyldisilazane, immediately centrifuged and the pellet deposited onto clean glass coverslips. Specimens were dried at 60°C, sputtered with ca. 350 A of palladium/gold and imaged in a Hitachi S570 SEM.


Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 987-997 ◽  
Author(s):  
Bryan Kindiger

Cytogenetic investigations of meiosis in hybrids between maize and Tripsacum have been well documented; however, the inherent problem of male and female sterility has not been addressed either on a genetic or cytogenetic level. The purpose of this cytological study was to identify some of the probable causes of male sterility in maize × Tripsacum dactyloides hybrids. Disturbances in pollen development of maize × T. dactyloides hybrids, derived from both diploid (2n) and tetraploid (4n) Tripsacum sources, were commonly observed. Anomalies in the development of the microspore apparently occurred because of a failure of the chromosomes to congregate at the metaphase plate, development of a tripolar spindle, and failure of cytokinesis at the first and second meiotic divisions. Phenotypic features of abnormal microspore development were the maturation of large pollen grains, "Siamese" pollen grains, the occurrence of variable invaginations, and a nuclear budding-type behavior. These abnormalities were not observed in the 56-chromosome amphidiploid or the 38-chromosome backcross generations.Key words: maize, Tripsacum, microspore, sterility.


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