Elongation Of Segments Research Articles

Differential expression profiles of growth-related genes in the elongation zone of maize primary roots

Growth in the apical elongation zone of plant roots is central to the development of functional root systems. Rates of root segmental elongation change from accelerating to decelerating as cell development proceeds from newly formed to fully elongated status. One of the primary variables regulating these changes in elongation rates is the extensibility of the elongating cell walls. To help decipher the complex molecular mechanisms involved in spatially variable root growth, we performed a gene identification study along primary root tips of maize (Zea mays) seedlings using suppression subtractive hybridization (SSH) and candidate gene approaches. Using SSH we isolated 150 non-redundant cDNA clones representing root growth-related genes (RGGs) that were preferentially expressed in the elongation zone. Differential expression patterns were revealed by Northern blot analysis for 41 of the identified genes and several candidate genes. Many of the genes have not been previously reported to be involved in root growth processes in maize. Genes were classified into groups based on the predicted function of the encoded proteins: cell wall metabolism, cytoskeleton, general metabolism, signaling and unknown. In-situ hybridization performed for two selected genes, confirmed the spatial distribution of expression shown by Northern blots and revealed subtle differences in tissue localization. Interestingly, spatial profiles of expression for some cell wall related genes appeared to correlate with the profile of accelerating root elongation and changed appropriately under growth-inhibitory water deficit.

The spatially variable inhibition by water deficit of maize root growth correlates with altered profiles of proton flux and cell wall pH.

Growth of elongating primary roots of maize (Zea mays) seedlings was approximately 50% inhibited after 48 h in aerated nutrient solution under water deficit induced by polyethylene glycol 6000 at -0.5 MPa water potential. Proton flux along the root elongation zone was assayed by high resolution analyses of images of acid diffusion around roots contacted for 5 min with pH indicator gel. Profiles of root segmental elongation correlated qualitatively and quantitatively (r(2) = 0.74) with proton flux along the surface of the elongation zone from water-deficit and control treatments. Proton flux and segmental elongation in roots under water deficit were remarkably well maintained in the region 0 to 3 mm behind the root tip and were inhibited from 3 to 10 mm behind the tip. Associated changes in apoplastic pH inside epidermal cell walls were measured in three defined regions along the root elongation zone by confocal laser scanning microscopy using a ratiometric method. Finally, external acidification of roots was shown to specifically induce a partial reversal of growth inhibition by water deficit in the central region of the elongation zone. These new findings, plus evidence in the literature concerning increases induced by acid pH in wall-extensibility parameters, lead us to propose that the apparently adaptive maintenance of growth 0 to 3 mm behind the tip in maize primary roots under water deficit and the associated inhibition of growth further behind the tip are related to spatially variable changes in proton pumping into expanding cell walls.

Characterization of the concentration gradient of prostaglandin H synthase 2 mRNA throughout the pregnant baboon uterus.

The present study was designed to determine the effect of the spatial gradient from the cervix to the uterine fundus on the control of local prostaglandin H synthase (PGHS) 2 mRNA expression. We performed total cesarean hysterectomies during the last trimester in 12 pregnant baboons, 7 not in labor and 5 in labor, and examined PGHS2 mRNA expression throughout the uterus. PGHS2 mRNA abundance was quantified by in situ hybridization and northern blot analysis in the uterine fundus, lower uterine segment and the different segments of the cervix. Quantitative northern blot and in situ analysis demonstrated a gradient of PGHS2 mRNA expression, with the highest levels at the level of the lower portion of the cervix and decreased expression through the mid- and upper portion of the cervix and lower uterine segment; the lowest levels of expression were seen in the uterine fundus. Moreover, cellular localization of PGHS2 mRNA and protein demonstrated high levels of expression in the cervical glandular epithelial cells with only occasional staining of smooth muscle cells in pregnant baboons. Decreased PGHS2 mRNA concentration gradient from the cervical external os to the fundus suggests that prostaglandin (PG) production in the uterus and cervix strongly depends on anatomical relations. This increased local PG production activity may be critical to pregnancy-associated lower uterine segment elongation, cervical softening and effacement in primate labor. These data provide a compelling biological basis for the use of PGHS2 inhibitors in the prophylaxis of preterm birth and cervical incompetence.

Towards a revision of the genus Parastenocaris Kessler, 1913: establishment of Simplicaris gen. nov. from groundwaters in central Italy and review of the P. brevipes -group (Copepoda, Harpacticoida, Parastenocarididae)

A new genus Simplicaris (Copepoda, Harpacticoida, Parastenocarididae) is established to accommodate two species collected from deep groundwater in Italy, S. lethaea sp. nov. and S. veneris (Cottarelli & Maiolini, 1980) comb. nov.Parastenocaris hippurisHertzog, 1938 and P. aedesHertzog, 1938 are ranked as incertae sedis within the genus. Members display complete absence of leg 5 in both sexes and an unusual elongation of the first exopodal segments of legs 1–4, in which exp-1 is distinctly longer than exp-2 or -3, or as long as exp-2 and -3 combined. As the systematic status of the family Parastenocarididae and of the type genus Parastenocaris is still in flux, a list of phylogenetically informative characters is proposed, along with a discussion of their various states in representative members of the family. The genus Parastenocaris sensu stricto is redefined to comprise only the brevipes-group. The remaining members of the genus are considered Parastenocaris sensu lato pending a thorough revision of the family.

The Auxin-induced Maize Gene ZmSAUR2 Encodes a Short-lived Nuclear Protein Expressed in Elongating Tissues

By differential screening of a cDNA library from auxin-induced maize coleoptiles we have isolated and characterized a SAUR gene, designated ZmSAUR2, belonging to a not yet characterized subtype of the SAUR family. ZmSAUR2 encodes a 15.3-kDa protein and is specifically induced by auxin in elongating coleoptile tissue but not in primary leaves or in roots. The transcript level rapidly increased within minutes and preceded auxin-stimulated elongation of coleoptile segments. Cycloheximide also induced ZmSAUR2 transcription, as has been shown for other early auxin-induced genes, whereas abscisic acid, brassinolide, ethylene, gibberellic acid, kinetin, and methyl jasmonate did not provoke an increase in ZmSAUR2 mRNA abundance. In pulse-chase experiments using auxin-induced coleoptiles and an anti-ZmSAUR2 antibody we were able to precipitate a protein of the expected molecular mass and to determine a half-life of about 7 min, which is among the shortest known in eukaryotes. In gel shift assays binding of calmodulin to ZmSAUR2 was demonstrated, suggesting the possibility of post-transcriptional regulation. Upon transformation of onion epidermal cells with a ZmSAUR2::GUS construct the corresponding chimeric protein was detected in the nucleus. The results suggest that ZmSAUR2 encodes a short-lived nuclear protein that might be involved in auxin-mediated cell elongation.

Interactions between abscisic acid, ethylene and gibberellin in internodal elongation in floating rice: the promotive effect of abscisic acid at low humidity

The enhancement of internodal elongation in floating or deepwater rice (Oryza sativa L. cv. Habiganj Aman II) by treatment with ethylene or gibberellic acid (GA3) at high relative humidity (RH) is inhibited by abscisic acid (ABA). Here, we examined the interactive effects of ethylene, gibberellin (GA) and ABA at low RH on internodal elongation of deepwater rice stem segments. Although ethylene alone hardly promoted internodal elongation of stem sections at 30% RH, it enhanced the internodal elongation induced by GA3. Application of ABA alone to stem segments had no effect on internodal elongation. However, in the presence of ethylene and GA3 at 30% RH, ABA further promoted internodal elongation. This promotive effect of ABA was not found in the internodes of stem segments treated either with ethylene or with GA3 at 30% RH or in the internodes of stem segments treated with ethylene and/or GA3 at 100% RH.

Post-hatch development of dendritic arborization in cerebellar Purkinje neurons of quail chicks: a morphometric study

To study development of the cerebellum in precocial birds during the early post-hatch period, dendritic arborization of Purkinje neurons was analyzed by intracellular Lucifer-yellow filling in fixed slices. Purkinje neurons were sampled from parasagittal slices of the cerebellar vermis of quail chicks (from 1 day pre-hatch to 14 days post-hatch). Confocal images revealed that the dendritic arborization expanded two-fold during the initial 3 days post-hatch, whereas the growth of cell bodies was much smaller. The dendritic expansion was accompanied by significant elongation of distal dendritic segments (5th and more distal segments), while the length of proximal dendrites (1st–4th segments) and the number of dendritic branches remained unchanged. Active synaptogenesis may occur selectively in distal dendrites during the early post-hatch period.

Reactive oxygen species in the elongation zone of maize leaves are necessary for leaf extension.

The production and role of reactive oxygen species (ROS) in the expanding zone of maize (Zea mays) leaf blades were investigated. ROS release along the leaf blade was evaluated by embedding intact seedlings in 2',7'-dichlorofluorescein-containing agar and examining the distribution of 2',7'-dichlorofluorescein fluorescence along leaf 4, which was exposed by removing the outer leaves before embedding the seedling. Fluorescence was high in the expanding region, becoming practically non-detectable beyond 65 mm from the ligule, indicating high ROS production in the expansion zone. Segments obtained from the elongation zone of leaf 4 were used to assess the role of ROS in leaf elongation. The distribution of cerium perhydroxide deposits in electron micrographs indicated hydrogen peroxide (H(2)O(2)) presence in the apoplast. 2',7'-Dichlorofluorescein fluorescence and apoplastic H(2)O(2) accumulation were inhibited with diphenyleneiodonium (DPI), which also inhibited O*(2)(-) generation, suggesting a flavin-containing enzyme activity such as NADPH oxidase was involved in ROS production. Segments from the elongation zone incubated in water grew 8% in 2 h. KI treatments, which scavenged H(2)O(2) but did not inhibit O*(2)(-) production, did not modify growth. DPI significantly inhibited segment elongation, and the addition of H(2)O(2) (50 or 500 microM) to the incubation medium partially reverted the inhibition caused by DPI. These results indicate that a certain concentration of H(2)O(2) is necessary for leaf elongation, but it could not be distinguished whether H(2)O(2), or other ROS, are the actual active agents.

Defects in the MITFmi / miApical Surface are Associated with a Failure of Outer Segment Elongation

The loss of MITF function in the MITFmi / mimouse affects not only RPE differentiation, but also the development of rod photoreceptor outer segments. Our data indicate that opsin immunoreactivity is detected in the cell membrane and along the ONL/RPE border of developing MITFmi / mirod photoreceptors and that rod outer segment morphogenesis is initiated. Although molecules associated with the outer segment continued to be expressed, outer segments did not elongate and develop stacked organized discs perpendicular to the RPE. The MITFmi /miRPE also failed to form apical microvilli and lacked the apical network of the phosphoprotein ezrin seen in wild type tissue. The MITFmi / miRPE basal surface was loosely organized and retained ezrin labelling which indicated some degree of differentiation. The correlation seen in our data suggest that there may be a link between the failure of the RPE apical domain to form and lack of rod outer segment elongation.

Gibberellic acid and dwarfism effects on the growth dynamics of B73 maize ( Zea mays L.) leaf blades: a transient increase in apoplastic peroxidase activity precedes cessation of cell elongation

The relationship between apoplastic peroxidase (EC 1.11.1.7) activity and cessation of growth in maize ( Zea mays L.) leaf blades was investigated by altering elongation zone length. Apoplastic peroxidase activity in the elongation and secondary cell wall deposition zones of elongating leaf blades of the maize inbred line B73 was used as a control and compared to leaves of the dwarf mutant D8‐81127, a near‐isogenic line of B73 unresponsive to gibberellins, and to leaves of B73 plants to which gibberellic acid (GA 3 ) had been applied via root uptake. Elongation zone length was increased by treatment with GA 3 through an increase in cell number as well as increased final cell length. The shorter elongation zone of dwarf leaves occurred primarily through reduced final cell length. Although elongation zone length differed among dwarf, control, and GA 3 ‐treated leaf blades, in all three treatments a transient increase in apoplastic peroxidase activity preceded a reduction in the segmental elongation rate in leaves. A peroxidase isoenzyme with pI 7.0 occurred in the leaf elongation zone during growth deceleration in all three treatments, and its activity decreased as growth displaced tissue into the region of secondary cell wall deposition. Growth cessation for all treatments coincided with the first appearance of peroxidase isozymes with pIs of 5.6 and 5.7. Based on the activity of particular isozymes relative to growth and differentiation, the pI 7.0 isoenzyme is most likely to be involved in cessation of cell elongation, while isozymes with pIs 5.6 and 5.7 are likely to be active in lignification.

Auxin-induced elongation of short maize coleoptile segments is supported by 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one.

Endogenous extractable factors associated with auxin action in plant tissues were investigated, especially their effects on elongation of 1-mm coleoptile segments of maize (Zea mays L.), in the presence of saturating 10 microM indole-3-acetic acid (IAA). The relative growth response, to auxin alone, was much smaller in segments shorter than 2-3 mm compared to 10-mm segments. Fusicoccin-induced elongation, however, was less affected by shortening the segments. A reduced auxin response may result from the depletion through cut surfaces of a substance required for IAA-mediated growth. Sucrose, phenolics like flavonoids, and vitamins were ruled out as the causal factors. A partially purified methanol extract of maize coleoptiles supported longterm, auxin-controlled elongation. The active material was also found among substances bleeding from scrubbed maize coleoptiles. The active factor from maize was further purified by HPLC and characterised by the UV spectrum and its pH shift. This factor was identified as 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) by mass spectroscopy. Activity tests confirmed that pure DIMBOA from other sources sustained auxin-induced elongation of short maize coleoptile segments. However, DIMBOA only partially restored the activity lost from short segments. This indicates that an additional factor, other than DIMBOA, is required. Extracts from Avena or Cucurbita did not contain the factor DIMBOA; it was active on maize elongation, but not on Avena coleoptiles or Cucurbita hypocotyls. This narrow specificity and the lack of DIMBOA action in short-term tests with maize indicate that DIMBOA is not the general auxin cofactor but may specifically "spare" the co-auxin in maize.

Involvement of cortical microtubules in plastic extension regulated by gibberellin in Lemna minor root.

We aimed to analyze the rheological characteristics during elongation of the root segments in Lemna minor. The elastic component of segment elongation (EC) increased for the first 6 h, and then almost stopped. However, the plastic component of the segment elongation (PC) began to rapidly increase from 6 h onwards. Uniconazole-P, a gibberellin biosynthesis inhibitor, inhibited the total elongation of root segments (TE), and this inhibition was mainly caused by suppression of the rapid increase in the PC after 6 h. Concomitant with this inhibition, the cortical microtubule (CMT) array within root epidermal cells became disorganized in the presence of uniconazole-P from 6 h onwards. Adding GA3 abolished the inhibition of TE by uniconazole-P treatment, and this recovery was caused not by the increase in the EC but by an increase in the PC. Furthermore, the CMT arrays also recovered their characteristic organization in the presence of GA3. These findings suggest that endogenous gibberellin accelerates TE by activating the PC via control of CMT arrays. This conclusion is also supported by rheological analysis where propyzamide was used to disrupt microtubules. We suggest that endogenous gibberellin controls the PC via its influence over the transverse arrangement of CMTs.

Structure formation of a single polymer chain. I. Growth of trans domains

Molecular dynamics simulations are carried out to study structure formation of a single polymer chain with 500 CH2 groups. Our simulations show that the orientationally ordered structure is formed at a low temperature both by gradual stepwise cooling and by quenching from a random configuration at a higher temperature. The growth of the global orientational order proceeds in a gradual manner in the case of gradual stepwise cooling, whereas it proceeds in a stepwise manner in the case of quenching. The latter feature endorses the previously proposed hypothetical grand view of self-organization [e.g., see T. Sato, Phys. Plasmas 3, 2135 (1996)]: when a system is driven far from equilibrium, it will evolve to a more stable state in a stepwise fashion irrespective of its fundamental interaction forces. From the microscopic analysis of the structure formation process, we find the following characteristic features: (i) In the case of gradual stepwise cooling, the global orientational order grows gradually through the incorporation of small trans domains and the surrounding trans segments into the largest trans domain. (ii) In the case of quenching, the growth of the orientational order is either due to the incorporation of small trans domains and the surrounding trans segments into the largest trans domain or to the elongation of the trans segments in the largest trans domain.

Flower stalk segments of Arabidopsis thaliana ecotype Columbia lack the capacity to grow in response to exogenously applied auxin.

Exogenously applied IAA stimulated cell elongation of segments excised from flower stalks of Arabidopsis thaliana ecotype Landsberg erecta (Ler) by increasing the cell wall extensibility, but it did not affect that of ecotype Columbia (Col). Treatment with a low pH buffer solution (pH 4.0) or fusicoccin (FC), a reagent activating H(+)-ATPases, significantly increased the cell wall extensibility and promoted elongation growth of flower stalk segments of both ecotypes, indicating that the flower stalk segments of Col possess the capacity to grow under acidic pH conditions. IAA promoted the proton excretion in segments of Ler but not of Col. On the other hand, FC increased the proton excretion in segments of Col as much as that of Ler. These results suggest that IAA activates the plasma membrane H(+)-ATPases in the segments of Ler but not those of Col, while FC activates them in both ecotypes. Flower stalks of Col may lack the mechanisms of activation by IAA of the plasma membrane H(+)-ATPases.

A new concept of the significance of regional distribution of prostaglandin H synthase 2 throughout the uterus during late pregnancy: Investigations in a baboon model

Objective: We sought to identify regional differences in prostaglandin H synthase 2 messenger ribonucleic acid expressions in various intrauterine tissues in the pregnant baboon as an indicator of prostaglandin production capability to explain the various interactive roles of different intrauterine tissues in the processes that precede, promote, and complete labor. Study Design: Prostaglandin H synthase 2 messenger ribonucleic acid expression was measured by reverse transcriptase–polymerase chain reaction or Northern blot analysis in the uterine fundus, lower uterine segment, cervix, amnion, chorion, and placenta during late pregnancy and spontaneous term labor in the pregnant baboon. Myometrial electromyography enabled clear relation of the findings to uterine contractile activity. Results: There were dramatic increases of prostaglandin H synthase 2 messenger ribonucleic acid expressions during late gestation and during labor in the lower uterine segment, cervix, and decidua. The amniotic prostaglandin H synthase 2 messenger ribonucleic acid expression increased during labor. In contrast, the prostaglandin H synthase 2 messenger ribonucleic acid expressions in the uterine fundus, chorion, and placenta did not change during late gestation and labor. Conclusion: Demonstrated increased lower uterine segment and cervical prostaglandin H synthase 2 abundances would promote lower uterine segment elongation and cervical effacement. Engagement of the fetal presenting part would stimulate local prostaglandin H synthase 2 expression and obstruct diffusion of high forebag prostaglandin to the rest of the uterus, as reported previously in human pregnancy. These data support a new conceptual mechanistic framework for preparatory changes in the lower uterine segment and cervix preceding labor as precisely related to myometrial contractility changes. (Am J Obstet Gynecol 2000;183:1287-95.)

Regulation of elongation growth by gibberellin in root segments of Lemna minor.

Hormonal control of elongation growth was analyzed in segments excised from the elongation zone of Lemna roots. Exogenous GA3 did not promote the segment elongation but rather inhibited it. Uniconazole-P, a gibberellin biosynthesis inhibitor, significantly inhibited the segment elongation, and the inhibitory effect was completely nullified by GA3. In the epidermis, cell elongation was inhibited, but lateral cell expansion was not affected by uniconazole-P. Orientation of cortical microtubules of epidermal cells was disturbed by treatment with uniconazole-P for 12 h, and the disorganization of cortical microtubules was ameliorated by GA3. These findings suggested that disorganization of cortical microtubules induced inhibition of elongation growth of root. However, stabilization of cortical microtubules by taxol, a microtubule-stabilizing agent, did not affect the inhibition of segment elongation by uniconazole-P. These results suggested that endogenous gibberellin controls the elongation growth of root by regulating cell elongation.

Choline conjugates of auxins. I. Direct evidence for the hydrolysis of choline-auxin conjugates by pea cholinesterase

We synthesized the choline conjugates of three derivatives of phenylacetic acid, four derivatives of phenoxyacetic acid, and of naphthalene-1-acetic acid and measured the effects of these conjugates on pea ( Pisum sativum L. cv. Alaska) stem segment elongation. We also synthesized the thioester analogs of six choline conjugates and measured their rates of hydrolysis by pea cholinesterase and their rates of spontaneous hydrolysis. With one exception, conjugates that stimulated growth were hydrolyzed at high rates by pea cholinesterase, while conjugates that were hydrolyzed at low rates by pea cholinesterase did not stimulate growth. The results are consistent with a model in which cholinesterase releases active auxins from these conjugates.

Disruption of Laminin β2 Chain Production Causes Alterations in Morphology and Function in the CNS

From the elegant studies of Ramon y Cajal (1909) to the current advances in molecular cloning (e.g., Farber and Danciger, 1997), the retina has served as an ideal model for the entire CNS. We have taken advantage of the well described anatomy, physiology, and molecular biology of the retina to begin to examine the role of the laminins, one component of the extracellular matrix, on the processes of neuronal differentiation and synapse formation in the CNS. We have examined the effect of the deletion of one laminin chain, the beta2 chain, on retinal development. The gross development of retinas from laminin beta2 chain-deficient animals appears normal, and photoreceptors are formed. However, these retinas exhibit several pathologies: laminin beta2 chain-deficient mice display abnormal outer segment elongation, abnormal electroretinograms, and abnormal rod photoreceptor synapses. Morphologically, the outer segments are reduced by 50% in length; the outer plexiform layer of mutant animals is disrupted specifically, because only 7% of observed rod invaginating synapses appear normal, whereas the inner plexiform layer is undisturbed; finally, the rate of apoptosis in the mutant photoreceptor layer is twice that of control mice. Physiologically, the electroretinogram is altered; the amplitude of the b-wave and the slope of the b-wave intensity-response function are both decreased, consistent with synaptic disruption in the outer retina. Together, these results emphasize the prominence of the extracellular matrix and, in particular, the laminins in the development and maintenance of synaptic function and morphogenesis in the CNS.

Al Binding in the Epidermis Cell Wall Inhibits Cell Elongation of Okra Hypocotyl

Al inhibits root elongation at micromolar concentrations, but the mechanisms leading to this process are unknown. In these studies, Al-induced inhibition of cell elongation was examined using hypocotyl of okra (Abelmoschus esculentus Moench cv. Clemson Spineless) as an experimental model. One-h exposure to Al (0.5 mM A1CI3) in the presence of 10 /uM auxin in 0.5 mM CaCI2, pH 4.0 significantly inhibited auxin-induced cell elongation of okra hypocotyl segments. Elongation was further suppressed with increasing Al concentration s up to 1 mM. Treatment of the hypocotyl with 1 mM citrate for 10 minutes after 2-h exposure to Al resulted in significant recovery of elongation. The amount of Al in the cell wall relative to the total in the tissue was 96.0,96.2, and 85.4%, respectively, following 1-, 2-, and 3-h exposure to the Al solution. The total and cell wall Al content was decreased by half after the citrate desorption treatment. Furthermore, 95% of Al was found in the epidermis, and 95% of the Al in the epidermis was associated with the cell wall. Experiments using split hypocotyl segments showed that Al exposure increased the outward bending of hypocotyl segments, suggesting that the epidermis elongation was specifically inhibited by Al. Al inhibited the autolysis of epidermis by about 20%, but had little effect on the autolysis of core tissue. Taken together, these results suggest that Al binding in the epidermal cell wall inhibits critical components in cell wall loosening mechanism, resulting in inhibition of cell elongation.

Occurrence of cello-oligosaccharides in the apoplast of auxin-treated pea stems

Treatment of pea (Pisum sativum L.) hypocotyl segments with indole-3-butyric acid, which promotes segment elongation, increased the solubilization of both xyloglucan and cello-oligosaccharides in the apoplast of auxin-treated pea stems. The cello-oligosaccharides were isolated from the apoplastic solution with a charcoal/Celite column and were identified as cellobiose, cellotriose, and cellotetraose after subsequent thin-layer chromatography and paper electrophoresis. Cello-oligosaccharides in the apoplastic fraction were monitored using cellobiose dehydrogenase. Both xyloglucan and cello-oligosaccharides appeared to be formed concurrently within 30 min after treatment with the auxin, and the cello-oligosaccharides increased with stem elongation even after 2 h. The total activity of cellulase did not increase for up to 4 h.