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PubMed2000

An increase in pectin methyl esterase activity accompanies dormancy breakage and germination of yellow cedar seeds.

Pectin methyl esterase (PME) (EC 3.1.1.11) catalyzes the hydrolysis of methylester groups of cell wall pectins. We investigated the role of this enzyme in dormancy termination and germination of yellow cedar (Chamaecyparis nootkatensis [D. Don] Spach) seeds. PME activity was not detected in dormant seeds of yellow cedar but was induced and gradually increased during moist chilling; high activity coincided with dormancy breakage and germination. PME activity was positively correlated to the degree of dormancy breakage of yellow cedar seeds. The enzyme produced in different seed parts and in seeds at different times during moist chilling, germination, and early post-germinative growth consisted of two isoforms, both basic with isoelectric points of 8.7 and 8.9 and the same molecular mass of 62 kD. The pH optimum for the enzyme was between 7.4 and 8.4. In intact yellow cedar seeds, activities of the two basic isoforms of PME that were induced in embryos and in megagametophytes following dormancy breakage were significantly suppressed by abscisic acid. Gibberellic acid had a stimulatory effect on the activities of these isoforms in embryos and megagametophytes of intact seeds at the germinative stage. We hypothesize that PME plays a role in weakening of the megagametophyte, allowing radicle emergence and the completion of germination.

Abscisic Acid↗

PubMed2001

Proteomic analysis of arabidopsis seed germination and priming.

To better understand seed germination, a complex developmental process, we developed a proteome analysis of the model plant Arabidopsis for which complete genome sequence is now available. Among about 1,300 total seed proteins resolved in two-dimensional gels, changes in the abundance (up- and down-regulation) of 74 proteins were observed during germination sensu stricto (i.e. prior to radicle emergence) and the radicle protrusion step. This approach was also used to analyze protein changes occurring during industrial seed pretreatments such as priming that accelerate seed germination and improve seedling uniformity. Several proteins were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry. Some of them had previously been shown to play a role during germination and/or priming in several plant species, a finding that underlines the usefulness of using Arabidopsis as a model system for molecular analysis of seed quality. Furthermore, the present study, carried out at the protein level, validates previous results obtained at the level of gene expression (e.g. from quantitation of differentially expressed mRNAs or analyses of promoter/reporter constructs). Finally, this approach revealed new proteins associated with the different phases of seed germination and priming. Some of them are involved either in the imbibition process of the seeds (such as an actin isoform or a WD-40 repeat protein) or in the seed dehydration process (e.g. cytosolic glyceraldehyde-3-phosphate dehydrogenase). These facts highlight the power of proteomics to unravel specific features of complex developmental processes such as germination and to detect protein markers that can be used to characterize seed vigor of commercial seed lots and to develop and monitor priming treatments.

Arabidopsis↗

PubMed2005

A new role for the Arabidopsis AP2 transcription factor, LEAFY PETIOLE, in gibberellin-induced germination is revealed by the misexpression of a homologous gene, SOB2/DRN-LIKE.

Gibberellic acid (GA) promotes germination, stem/hypocotyl elongation, and leaf expansion during seedling development. Using activation-tagging mutagenesis, we identified a mutation, sob2-D (for suppressor of phytochromeB-4 [phyB-4]#2 dominant), which suppresses the long-hypocotyl phenotype of a phyB missense allele, phyB-4. This mutant phenotype is caused by the overexpression of an APETALA2 transcription factor, SOB2, also called DRN-like. SOB2/DRN-like transcript is not detectable in wild-type seedling or adult tissues via RT-PCR analysis, suggesting that SOB2/DRN-like may not be involved in seedling development under normal conditions. Adult sob2-D phyB-4 plants have curled leaves and club-like siliques, resembling plants that overexpress a closely related gene, LEAFY PETIOLE (LEP). Hypocotyls of a LEP-null allele, lep-1, are shorter in the light and dark, suggesting LEP involvement in seedling development. This aberrant hypocotyl phenotype is due at least in part to a delay in germination. In addition, lep-1 is less responsive to GA and more sensitive to the GA biosynthesis inhibitor paclobutrazol, indicating that LEP is a positive regulator of GA-induced germination. RT-PCR shows that LEP transcript accumulates in wild-type seeds during imbibition and germination, and the transcript levels of REPRESSOR OF ga1-3-LIKE2 (RGL2), a negative regulator of GA signaling during germination, is unaffected in lep-1. These results suggest LEP is a positive regulator of GA-induced germination acting independently of RGL2. An alternative model places LEP downstream of RGL2 in the GA-signaling cascade.

Amino Acid Sequence↗

PubMed2006

Tomato seed germination: regulation of different response modes by phytochrome B2 and phytochrome A.

Lycopersicon esculentum seeds germinate after rehydration in complete darkness. This response was inhibited by a far-red light (FR) pulse, and the inhibition was reversed by a red light (R) pulse. Comparison of germination in phytochrome-deficient mutants (phyA, phyB1, phyB2, phyAB1, phyB1B2 and phyAB1B2) showed that phytochrome B2 (PhyB2) mediates both responses. The germination was inhibited by strong continuous R (38 micromol m(-2) s(-1)), whereas weak R (28 nmol m(-2) s(-1)) stimulated seed germination. Hourly applied R pulses of the same photon fluence partially replaced the effect of strong continuous R. This response was called 'antagonistic' because it counteracts the low fluence response (LFR) induced by a single R pulse. This antagonistic response might be an adaptation to a situation where the seeds sit on the soil surface in full sunlight (adverse for germination), while weak R might reflect that situation under a layer of soil. Unexpectedly, the effects of continuous R or repeated R pulses were mediated by phytochrome A (PhyA). We therefore suggest that low levels of PhyA in its FR-absorbing form (Pfr) cause inhibition of seed germination produced either by extended R irradiation (by degradation of PhyA-Pfr) or by extended FR irradiation [keeping a low Pfr/R-absorbing form (Pr) ratio].

Germination↗

PubMed1996

The role of MT2 and anti-bcl-2 protein antibodies in the differentiation of benign from malignant cutaneous infiltrates of B-lymphocytes with germinal center formation.

MT2/CD45RA and anti-bcl-2 protein (Bcl-2) monoclonal antibodies are useful markers in distinguishing follicular lymphomas from reactive follicular hyperplasia of the lymph nodes. We examined biopsy specimens from 11 patients with primary cutaneous B-cell follicle center lymphomas, 10 patients with cutaneous pseudolymphomas with germinal centers, and 6 patients with inflammatory infiltrates with germinal centers in non-lymphoid cutaneous tumors (3 basal cell carcinomas, 2 malignant melanomas, and 1 solar keratosis), in order to evaluate the utility of MT2 and anti-Bcl-2 antibodies in differentiating benign from malignant germinal center cell proliferations in the skin. Immunohistochemical evaluation of MT2 and Bcl-2 was focused exclusively on the reactivity of germinal center cells. Specific membranous MT2 positivity was found in 2/11 cutaneous follicle center lymphomas; a diffuse, non-specific staining pattern was identified in 3/11 follicle center lymphomas and in 1/6 inflammatory infiltrates in non-lymphoid tumors. A negative MT2 reaction was observed in 6/11 follicle center cell lymphomas, in all cases of pseudolymphomas and in 5/6 inflammatory infiltrates in non-lymphoid tumors. Bcl-2 positivity was detected only in 1/11 follicle center lymphomas. Germinal center cells in all other cases were Bcl-2 negative. Our results suggest that MT2 and anti-Bcl-2 antibodies are only of limited value in differentiating primary cutaneous follicle center lymphomas from cutaneous pseudolymphomas with germinal centers.

Antibodies, Monoclonal↗

PubMed1997

Germinal center development.

Using a set of surface markers including IgD and CD38, human tonsillar B cells were classified into discrete subpopulations. Molecular and functional analysis allowed us to identify: i) two sets of naive B cells (Bm1 and Bm2); ii) germinal center founder cells (Bm2'); iii) an obscure population of germinal center B cells, displaying a high load of somatic mutations in IgV genes, C mu to C delta switch and preferential Ig lambda light chain usage: these cells may represent the precursors of normal and malignant IgD-secreting plasma cells; iv) the centroblasts (Bm3) in which somatic mutation machinery is activated; v) the centrocytes (Bm4) in which isotype switch occurs; vi) the memory B cells. The characterization of these subpopulations showed that: i) programmed cell death is set before somatic mutations, possibly providing an efficient way for affinity maturation; ii) only high affinity centrocytes are allowed to switch isotype; iii) CD40-ligation inhibits plasmacytic differentiation of mature B lymphocytes; iv) memory B cells preferentially differentiate into plasma cells; v) IgD isotype switch occurs in normal B cells; vi) receptor editing may be induced by somatic mutations in germinal centers. We also characterized two types of antigen-presenting cells in germinal centers: follicular dendritic cells that select high affinity B cells, and a new subset of germinal center dendritic cells that activate germinal center T cells.

Animals↗

PubMed1997

Lymphotoxin-alpha-deficient and TNF receptor-I-deficient mice define developmental and functional characteristics of germinal centers.

Mice deficient in LT alpha (LT alpha-/-) lack lymph nodes and Peyer's patches. This action of LT alpha in lymph node organogenesis appears to be mediated by the membrane form of LT using a mechanism independent of TNF receptor I (TNFR-I) or II (TNFR-II). In contrast, normal Peyer's patch development appears to require both LT alpha and TNFR-I, with TNFR-I-/- mice showing hypoplastic Peyer's patch structures. LT alpha-/- mice also fail to support the normal segregation of T-cell and B-cell zones within the splenic white pulp. Again, this occurs via a mechanism independent of TNFR-I or TNFR-II. Additionally, follicular dendritic cell (FDC) clusters or germinal centers fail to develop in the spleen of LT alpha-/- animals. Mice deficient in either TNF alpha or TNFR-I also fail to develop splenic FDC clusters and germinal centers, indicating that signaling by both LT alpha and TNF alpha is required for development of these specialized lymphoid tissue structures. Finally, the splenic white pulp areas in LT alpha-/- mice lack the marginal zone of monoclonal antibody MOMA-1-staining metallophilic macrophages, whereas TNFR-I-deficient mice have preserved MOMA-1 staining. Thus, certain actions of LT alpha to regulate spleen white pulp architecture are mediated by receptors other than TNFR-I, most likely by the LT beta R or a closely related receptor. We tested whether germinal centers are essential for maturation of T-cell-dependent antibody responses. When LT alpha-/- mice were immunized with low doses of NP-ovalbumin (NP-OVA) adsorbed to alum, there was dramatically impaired production of high affinity anti NP IgG; however, after immunization with high doses of NP-OVA adsorbed to alum, LT alpha-/- mice mounted a high affinity NP-specific serum IgG response similar to wild-type mice, all in the absence of germinal centers or clustered FDC. Thus, although germinal centers enhance the processes required for maturation of the humoral immune response, the mechanisms responsible for affinity maturation are not absolutely dependent on the presence of germinal centers.

Animals↗

PubMed2026

Maize ZmMYB59 inhibits post-germinative shoot and root elongation through ZmGA2ox3/10-mediated gibberellin catabolism.

Gibberellin (GA) promotes seed germination, but sustained or excessive GA signaling after germination can lead to aberrant root and shoot elongation. How GA homeostasis is transcriptionally restrained during post-germinative seedling development remains unclear. Using overexpression and gene-edited maize materials, we demonstrate that ZmMYB59 inhibits root and shoot elongation during post-germinative growth. Integrated RNA-Seq and CUT&Tag analyses identified the GA catabolism genes ZmGA2ox3 and ZmGA2ox10 as candidate direct targets of ZmMYB59. Hormone profiling analysis showed elevated bioactive GA1 and GA4 levels in the scutellum and aleurone layer cells of zmmyb59 mutants. Dual-luciferase assays, electrophoretic mobility shift assays, and ChIP-qPCR further confirmed that ZmMYB59 directly binds AC8 cis-elements in the ZmGA2ox3/10 promoters and activates their transcription. The zmga2ox3/10 double mutant, but neither single mutant, exhibited enhanced root and shoot elongation, accompanied by GA4 accumulation. This phenotype was suppressed by exogenous application of the GA biosynthesis inhibitor uniconazole. Transcriptomic and biochemical analyses further revealed enhanced starch degradation, reduced starch content, and increased soluble sugar accumulation in the double mutant. Taken together, these findings reveal that the ZmMYB59-ZmGA2ox3/10 module restrains GA accumulation and starch mobilization after germination, thereby coordinating reserve utilization with post-germinative root and shoot growth in maize.

Gibberellins↗

PubMed2001

Modulation of germination and growth of plants by meditation.

So called primitive peoples of the world share a philosophy that human interaction via ceremony or ritual can affect the natural world. Is it possible to affect the germination and growth of plants by imbuing them with an intent to stimulate or inhibit them? We conducted a double blind series of experiments to determine whether a process of meditation on the water (referred to as "treated") given to a controlled planting of green peas or wheat would affect their germination. Peas were given water treated with stimulating intent. Statistical analysis was done using contingency table, Fisher's test, and Mantel-Haenszel analysis. The germination rate of 504 seeds receiving treated water with stimulating intent was 60.3% compared to 51.8% for the 504 controls (p = 0.006, 0.047, 0.003 respectively). A similar experiment was conducted with wheat with the intent of inhibiting germination. The germination rate of 2970 wheat seeds receiving treated water with inhibitory intent was 70.7% versus 74.9% for 2970 controls (p < 0.001, 0.0001, 0.001 respectively). During the sixth run of the wheat (inhibition) experiment, the seedlings were harvested and individually weighed on the tenth day after planting to determine whether there was any difference in growth. The mass of the treated seedlings was statistically significantly lower (mean = 97 mg versus 106 mg for the controls) when compared by analysis of variance (p = 0.000056). We conclude that meditation upon the water supplied to green peas and wheat can affect their germination rates and growth.

Double-Blind Method↗

PubMed2005

Gibberellic acid nitrite stimulates germination of two species of light-requiring seeds via the nitric oxide pathway.

We used two species of light-requiring seeds, Paulownia tomentosa, which have absolute light requirement (no germination in darkness), and Stellaria media seeds, which germinate in darkness to a certain extent because of presence of preformed active phytochrome, to obtain results strongly suggesting that gibberellic acid nitrite stimulates seed germination via its capability as a functional NO donor. Exogenous application of gibberellic acid nitrite stimulates gibberellin-insensitive Stellaria media seed germination in darkness as do a wide variety of NO donors. Pure gibberellic acid could replace the light requirement of P. tomentosa seeds, thus enabling them to germinate in darkness. Gibberellic acid nitrite did not have this effect. A stimulative effect from gibberellic acid nitrite could be detected only after exposure of these seeds to short, 10 min, pulse of red light. Taken together, these results suggest that gibberellic activity of gibberellic acid nitrite is lost after nitrosation but, regarding to the presence of -O-NO moiety in the molecule, gibberellic acid nitrite shares stimulative properties in seed germination with other compounds with NO-releasing properties.

Dose-Response Relationship, Drug↗

PubMed1997

Preventive effect of germinated barley foodstuff on diarrhea induced by water-soluble dietary fiber in rats.

We investigated the preventive effect of germinated barley foodstuff (GBF) added to the diet on diarrhea induced by the dietary water-soluble dietary fibers, polydextrose, hemicellulose, and poly-acrylic acid sodium salt, in Sprague-Dawley rats. The minimum content of GBF necessary for blocking diarrhea was 3% (by weight) of the diet. Since GBF is mainly derived from the aleurone and scutellum of malted barley, we assessed the physiological effects of the aleurone and scutellum fractions derived from barley grains before and after germination. The addition of fractions containing only germinated barley, and not barley collected before germination, increased the fecal output and jejunal mucosal protein content. The effects of malted barley were very similar to those of GBF. It was concluded that germination was necessary to bring about the physiological effects of GBF. Since non-lignified hemicellulose and Gln-rich protein were newly synthesized during germination, these might have contributed to the increased fecal output and jejunal mucosal protein content.

Acrylic Resins↗

PubMed2004

Differential expression of acid invertase genes during seed germination in Arabidopsis thaliana.

In Arabidopsis thaliana (L.) Heynh ecotype Landsberg, levels of soluble acid invertase activity are closely related to the progress of seed germination. To study the mechanism(s) of the development of these enzymes, two cDNA clones that encode putative vacuolar acid invertases were isolated from germinating seeds and very young seedlings using reverse-transcription polymerase chain reactions with degenerate primers. These fragments corresponded to the genes At beta fruct3 and At beta fruct4 from the Columbia ecotype. An apoplasmic invertase gene corresponding to At beta fruct1/ATCWINV1 was also isolated from these samples. Northern blot analyses showed that At beta fruct3 and At beta fruc4 are expressed concomitantly with germination and the subsequent seedling growth. In contrast, the At beta fruct1/AtcwINV1 mRNA is translated before germination. These expression patterns are regulated by phytochrome, which perceives red light and in turn triggers de novo synthesis of gibberellin, initiating Arabidopsis seed germination. To test the effects of gibberellin on the expression of these genes, seed were treated with a gibberellin biosynthesis inhibitor, uniconazole or prohexadione. These chemicals inhibited both seed germination and expression of the above genes, but subsequently applied GA(4), an active gibberellin, reversed the inhibition. These results suggest that the transcription of genes encoding the vacuolar invertases, At beta fruct3 and At beta fruct4 and a gene encoding the apoplasmic invertase, At beta fruct1/AtcwINV1, are induced by gibberellin synthesized de novo following irradiation with red light.

Arabidopsis↗

PubMed2005

Degradation of soluble proteins including some allergens in brown rice grains by endogenous proteolytic activity during germination and heat-processing.

The effect of germination and subsequent heat-processing on the degradation of soluble proteins, including some allergenic proteins, in brown rice grains was investigated. The content of soluble proteins, including 14-16-kDa and 26-kDa allergens, in the germinated and processed brown rice grains (GPR) was much lower than that of non-germinated brown rice. These proteins in brown rice grains were also much lower after subsequent heat-processing during the manufacturing process. The protease activity of germinated brown rice (GR) was detected and increased 1.5 times after germination. The optimum pH values for degradation of the 26-kDa and 14-16-kDa allergens in the GR grains were 4 and between 5 and 7, respectively. These results suggest that the decrease in the soluble proteins and allergens was induced in part by proteolytic degradation. The presence of a detergent enhanced the proteolytic degradation of the soluble proteins, especially of the 26-kDa allergen, in the brown rice grains. The degradation of the 26-kDa allergen was weakly inhibited by NEM, suggesting cysteine protease(s) may have been involved in its degradation. These results suggest that the two abundant allergens were degraded in a different manner and probably by different proteases in the grains during germination.

Allergens↗

PubMed2003

Effects of simulated acid rain on the pollen germination and pollen tube growth of apple (Malus sylvestris Miller cv. Golden).

The pollens of apple flowers have been treated with simulated acid rain solutions in range of pHs 2.9 to 5.0 in order to determine the threshold proportion values that lead the observed symptoms of detriments of acid rain. Compared to controls (pH 6.5), pollen germination decreased by 41.75% at pH 3.3 and pollen tube elongation decreased by 24.3% at pH 3.4. Acid rain threshold proportion value was around pH 3.3 and 3.4 for apple pollen germination and pollen tube elongation, respectively. Furthermore, pollen tube elongation was determined to be more sensitive to acid rain than pollen germination. The pH values below 3.1 resulted in complete destruction of pollen tubes. Pollen germination entirely stopped at around pH 3.0. Finally, it has been shown that the acid rain has a blocking effect on pollen germination and pollen tube elongation in apple. The conclusion is that not only pH value but also the quantity of acid rain is important factor in germination. The results were found statistically significant through the LSD test at levels of p < 0.05 and p < 0.01.

Acid Rain↗

PubMed2000

Impaired affinity maturation in Cr2-/- mice is rescued by adjuvants without improvement in germinal center development.

Cr2-/- mice have an impairment in humoral immunity, as shown by the decrease in the Ab titers against T cell-dependent Ags and abnormalities in germinal center formation. Germinal centers are present, but they are decreased in size and number, indicating problems in their development. In this study, we investigated whether this abnormality in germinal center development is associated with problems in the establishment of optimal affinity maturation and the generation of memory B cells, processes closely related to the germinal center reaction. We immunized the Cr2-/- animals with different Ags with or without adjuvants. We showed that, when immunized without adjuvants, complement receptors are absolutely required for optimal affinity maturation. Although limited affinity maturation is elicited in the Cr2-/- Ab response, it is decreased as compared with normal animals. Memory B cell generation is also impaired. In the presence of adjuvants, germinal center development in the Cr2-/- mice is still abnormal, as demonstrated by their decreased size and number. Surprisingly, adjuvants establish optimal affinity maturation and partially restore the amount of Ab produced during the primary response and memory B cell generation. However, adjuvants cannot improve the ability of follicular dendritic cells to retain Ags in the form of immune complexes. These observations indicate that immunization with inflammatory Ags offset some of the immunological abnormalities found in the Cr2-/- mice and show that optimal affinity maturation in the Cr2-/- mice can be achieved in the absence of normal germinal centers.

Adjuvants, Immunologic↗

PubMed1996

No evidence of adverse effects on germination, emergence, and fruit yield due to space exposure of tomato seeds.

Seeds of 'Rutgers California Supreme' tomato (Lycopersicon esculentum Mill.) were exposed to outer space conditions aboard the long duration exposure facility (LDEF) satellite in the space exposed experiment developed for students (SEEDS) project of the National Aeronautics and Space Administration (NASA). Seeds aboard the LDEF were packed in dacron bags forming four layers per sealed canister. Some of these seeds were used in Oklahoma and Florida for studies of germination, emergence, and fruit yield. Among all measured variables in three experiments, there was only one significant main effect of canister 2 versus canister 7 (for mean time to germination) and only one main effect of layer (for seedling shoot dry weight). There also were only two inconsistent canister x layer interactions in the germination tests. The contrast of Earth-based control seed versus space-exposed seed was significant four times: in Oklahoma in 1991 the mean time to germination of space-exposed seeds and the days to 50% of final germination were 0.7 days less than for Earth-based seeds, and in Florida in 1992 seedling percent emergence and shoot dry weight were increased by space exposure. Fruit yield and marketability were unaffected in plants grown from space-exposed seeds. These results support student findings from the SEEDS project, and provide evidence that tomato seeds can survive in space for several years without adverse effects on germination, emergence, and fruit yield.

Cosmic Radiation↗

PubMed1998

[Metabolism in Arctium lappa L. at the primary stage of germination].

OBJECTIVE: To investigate the metabolic changes of the seeds at the primary stage of germination in Arctium lappa. METHOD: By means of electrophoresis the changes of peroxidases and soluble proteins were assayed at the primary stage of germination. RESULTS: The color of some original bands was enhanced, some original bands disappeared, and some new bands were detected in peroxidase patterns during the germinating process. The color changes were the main expression in soluble protein patterns. The bands of soluble proteins changed most obviously at 96 h and 120 h after germination. CONCLUSION: It follows that the metabolic process in the seeds of Arctium lappa at the stage of germination is complex and 96 h and 120 h after germination are the key times of metabolism.

Asteraceae↗

PubMed2002

Phytochrome E controls light-induced germination of Arabidopsis.

Germination of Arabidopsis seeds is light dependent and under phytochrome control. Previously, phytochromes A and B and at least one additional, unspecified phytochrome were shown to be involved in this process. Here, we used a set of photoreceptor mutants to test whether phytochrome D and/or phytochrome E can control germination of Arabidopsis. The results show that only phytochromes B and E, but not phytochrome D, participate directly in red/far-red light (FR)-reversible germination. Unlike phytochromes B and D, phytochrome E did not inhibit phytochrome A-mediated germination. Surprisingly, phytochrome E was required for germination of Arabidopsis seeds in continuous FR. However, inhibition of hypocotyl elongation by FR, induction of cotyledon unfolding, and induction of agravitropic growth were not affected by loss of phytochrome E. Therefore, phytochrome E is not required per se for phytochrome A-mediated very low fluence responses and the high irradiance response. Immunoblotting revealed that the need of phytochrome E for germination in FR was not caused by altered phytochrome A levels. These results uncover a novel role of phytochrome E in plant development and demonstrate the considerable functional diversification of the closely related phytochromes B, D, and E.

Apoproteins↗