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PubMed2000

Protection of tomato seed germination from the inhibitory effect of 2,4,5-trichlorophenoxyacetic acid by inoculation of soil with Burkholderia cepacia AC1100.

The effect of 2,4,5-trichlorophenoxy acetic acid (2,4,5-T) on the germination and seedling vigor of different crop seeds was tested. Seeds of rice, maize, sorghum, finger millet, and horse gram were comparatively more tolerant to the chemical with no marked effect up to a concentration of 200 mg 2,4,5-T L(-)(1) as tested by the filter paper method. Tomato and brinjal (egg plant) were highly susceptible. Even at 5 and 10 mg 2,4,5-T L(-)(1), marked reduction in the germination and seedling vigor of tomato and egg plant, respectively, was observed. At 20 and 30 mg L(-)(1), the germination of tomato and egg plant seeds, respectively, were completely inhibited on filter paper, whereas the inhibitory concentrations in soil was 40 mg 2,4,5-T kg(-)(1) soil. Several abnormalities were observed in the chemically affected seedlings. Protease activity of the seeds germinating in the presence of the chemical was drastically reduced. Bioremediation of the chemically contaminated soil with Burkholderia cepacia AC1100, by inoculation of the soil 7 days before sowing the seeds, completely protected the seeds, resulting in normal germination and an improved seedling vigor.

2,4,5-Trichlorophenoxyacetic Acid↗
PubMed2001

Nutritional quality of germinated cowpea flour (Vigna unguiculata) and its application in home prepared powdered weaning foods.

Amino acid profiles, protein digestibility, corrected amino acid scores (PDCAAS), chemical scores, essential amino acid indexes, and calculated biological values of controlcowpea flour (CCF), germinated cowpea flour (GCF) prepared from cowpeas germinated at 25 degrees C for either 24 h or 48 h and weaning foods prepared from cowpea flours were determined. Locally available rice, cowpea flour, banana-pumpkin slurry, and skim milk powder and sucrose in the ratio 35:35:15:15:5 were used to formulate weaning food containing not less than 15% protein. The ingredients were cooked into a slurry and oven-dried to produce flakes. The nutritional and sensory qualities of the weaning products were evaluated. Germination had little effect on the amino acid profile of cowpeas. In vitro protein quality and starch digestibility were improved in germinated cowpea flour. The PDCAAS of 24 h germinated cowpea flour (GCF) weaning food was higher (55.49%) than CCF-weaning food (46.74%). Vitamin A activity in 24 h GCF weaning food was higher than in CCF-weaning food. In vitro starch digestibilities of 24 h GCF and 48 h GCF-weaning foods were higher than that of CCF weaning food. The 24 h GCF-weaning food which had a higher overall acceptability score by sensory panelist than 48 h GCF and CCF-weaning food is recommended for household consumption.

Amino Acids↗
PubMed2003

Transport of Cl- across the plasma membrane during pollen grain germination in tobacco.

The role of Cl- transport across the plasma membrane was studied in an early step of pollen grain germination in tobacco Nicotiana tabacum L. The Cl- channel blockers, 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and niflumic acid, completely suppress the germination with IC(50) approximately 8 micro M. At this concentration NPPB reduces the rate of Cl- efflux out of pollen grain by 1.8-fold in the interval 5-12 min, and niflumic acid reduces the rate 1.2-fold. 4,4;-Diisothiocyanatostilbene-2,2;-disulfonic acid, a known inhibitor of Cl- channels and antiporters, completely suppresses germination as well (IC(50) = 240 micro M), but has no effect on the rate of Cl- efflux. Inhibitors of chloride co-transporters, such as furosemide, bumetanide, and bis(1,3-dibutylbarbituric acid)pentamethine oxonol, suppress the germination by less than 50%. This set of data suggests that NPPB-sensitive anion channels are involved in the activation of pollen grains in the early stage of germination.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Affinity maturation without germinal centres in lymphotoxin-alpha-deficient mice.

Affinity maturation by somatic hypermutation is thought to occur within germinal centres. Mice deficient in lymphotoxin-alpha (LT alpha-/- mice) have no lymph nodes or Peyer's patches, and fail to form germinal centres in the spleen. We tested whether germinal centres are essential for maturation of antibody responses to T-cell-dependent antigens. LT alpha-/- mice immunized with low doses of (4-hydroxy-3-nitrophenyl)acetyl-ovalbumin (NP-OVA) showed dramatically impaired production of high-affinity anti-NP IgG1. However, LT alpha-/- mice immunized with high doses of NP-OVA, even though they failed to produce germinal centres, manifested a high-affinity anti-NP IgG1 response similar to wild-type mice. Furthermore, when LT alpha-/- mice were multiply immunized with high doses of NP-OVA, the predominantly expressed anti-NP VH gene segment VH186.2 showed somatic mutations typical of affinity maturation. Thus, B-cell memory and affinity maturation are not absolutely dependent on the presence of germinal centres.

Amino Acid Sequence↗
PubMed2004

The BCL6 proto-oncogene suppresses p53 expression in germinal-centre B cells.

The human proto-oncogene BCL6 encodes a BTB/POZ-zinc-finger transcriptional repressor that is necessary for germinal-centre formation and is implicated in the pathogenesis of B-cell lymphoma. The precise function of BCL6 in germinal-centre development and lymphomagenesis is unclear because very few direct BCL6 target genes have been identified. Here we report that BCL6 suppresses the expression of the p53 (also known as tp53) tumour suppressor gene and modulates DNA damage-induced apoptotic responses in germinal-centre B cells. BCL6 represses p53 transcription by binding two specific DNA sites within the p53 promoter region and, accordingly, p53 expression is absent in germinal-centre B cells where BCL6 is highly expressed. Suppression of BCL6 expression via specific short interfering RNA leads to increased levels of p53 messenger RNA and protein both under basal conditions and in response to DNA damage. Most notably, constitutive expression of BCL6 protects B cell lines from apoptosis induced by DNA damage. These results suggest that an important function of BCL6 is to allow germinal-centre B cells to tolerate the physiological DNA breaks required for immunoglobulin class switch recombination and somatic hypermutation without inducing a p53-dependent apoptotic response. These findings also imply that deregulated BCL6 expression contributes to lymphomagenesis in part by functional inactivation of p53.

Apoptosis↗
PubMed1998

An evaluation of germinating beans as a source of vitamin C in refugee foods.

OBJECTIVE: To assess whether germinating pulses and beans are a potential source of vitamin C in rations for refugee communities with poor vitamin C status. DESIGN: High pressure liquid chromatographic assessment of vitamin C content of a range of legumes following germination in light and dark and after cooking. RESULTS: Many species of pulses produced significant quantities of vitamin C up to five days following germination in both light and dark although cooking caused a marked loss of ascorbate. CONCLUSION: Germination of approximately half of the seeds of the pulse constituents of many basic rations would be likely to generate, within a 3-5 d germination period, sufficient ascorbate to provide the 10 mg needed to protect adults against the development of scurvy.

Ascorbic Acid↗
PubMed2002

Lack of Shiga-like toxin binding sites in germinal centres of mouse lymphoid tissues.

B cells in germinal centres are known to express carbohydrate antigen CD77 in human lymphoid tissues. The CD77 antigen is specifically recognized by Shiga-like toxins (SLTs) that are produced by enterohaemorrhagic Escherichia coli O157:H7. To determine whether the binding subunits of Shiga-like toxin-1 (SLT-1B) could have adverse effects on the murine immune system when used as an immunogen, we investigated whether SLT-1B could bind to germinal centres of mouse lymphoid tissues. Frozen sections of peripheral lymph nodes and Peyer's patches from immunized mice were tested for the presence of SLT-1B-binding sites by immunohistological methods. Germinal centres were not stained with SLT-1B, while they were intensely stained with peanut agglutinin (PNA), another marker of germinal centres. On the other hand, SLT-1B specifically bound to renal tubules and collecting ducts in frozen sections of mouse kidney. This is consistent with results from human tissues. We also demonstrated that B220/PNA double-positive populations in lymph nodes from immunized mice exhibited only marginal staining with SLT-1B. The present results suggest that SLTs would not impede germinal centre functions of the murine immune system.

Animals↗
PubMed2002

Life and death within germinal centres: a double-edged sword.

Within germinal centres, B lymphocytes are destined to die by apoptosis via Fas signalling, unless they are positively rescued by antigen and by signals initiated by CD40-CD154 interactions. Thus, while the germinal centre microenvironment can become a virtual graveyard for most B lymphocytes that fail to bind antigen with high affinity, it concomitantly provides the necessary stimuli for the survival of cells that successfully accomplish affinity maturation. Such dichotomy in the physiology of germinal centre reaction that results in survival of the functional B-cell repertoire and the elimination of abnormal cells, dictates the fate towards B-cell homeostasis or disease. Consequently, the death and survival-signalling arms within germinal centres predominantly reside on the timely and controlled expression of Fas and its ligand (FasL), and CD40 and CD154, respectively. In keeping with this notion, lymphoproliferation or deficient immunity are documented landmarks of inactivation of either the Fas/FasL or CD40/CD154 signalling pathways. The present review considers two different scenarios in the control of B-cell survival and death within germinal centres. The first is an idealistic scenario, in which a discriminatory and co-ordinate signalling initiated by the CD40/CD154 and Fas/FasL pairs, respectively, leads the rescue of the functional B-cell repertoire and the elimination of the abnormal phenotype. The second is a gloomy scenario in which both the lack and the hyperexpression of either receptor/ligand pairs, are seen as equally deleterious.

B-Lymphocytes↗
PubMed2000

The Arabidopsis AtEPR1 extensin-like gene is specifically expressed in endosperm during seed germination.

Screening of 10 000 Arabidopsis transgenic lines carrying a gene-trap (GUS) construct has been undertaken to identify markers of seed germination. One of these lines showed GUS activity restricted to the endosperm, at the micropylar end of the germinating seed. The genomic DNA flanking the T-DNA insert was cloned by walking PCR and the insertion was shown to be located 70 bp upstream of a 2285 bp open reading frame (AtEPR1) sharing strong similarities with extensins. The AtEPR1 open reading frame consists of 40 proline-rich repeats and is expressed in both wild-type and mutant lines. The expression of the AtEPR1 gene appears to be under positive control of gibberellic acid, but is not downregulated by abscisic acid during seed germination. No expression was detected in organs other than endosperm during seed germination. The putative role of AtEPR1 is discussed in the light of its specific expression in relation to seed germination.

Amino Acid Sequence↗
PubMed2002

The symbiosis of Bacillus subtilis L-forms with Chinese cabbage seedlings inhibits conidial germination of Botrytis cinerea.

AIMS: To establish whether germination of Botrytis cinerea was affected by the symbiosis of Bacillus subtilis L-form bacteria with Chinese cabbage. METHODS AND RESULTS: Germinating seeds of Chinese cabbage were co-cultivated with either L-forms of Bacillus subtilis or 5% (w/v) mannitol by soaking for 3 h. Seeds were then washed in sterile water, sown on a minimal medium and incubated in controlled conditions. L-form symbiosis was detected over a time course by ELISA. Conidial germination of Botrytis cinerea was significantly reduced on cotyledonous leaves of L-form-treated plants compared with controls. CONCLUSIONS: Symbiosis of B. subtilis L-form bacteria during seed germination of Chinese cabbage inhibits conidial germination in plants on subsequent exposure to Botrytis cinerea. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first account of plant symbiosis with L-form bacteria showing antagonism to a fungal plant pathogen. This has promising implications for the use of this L-form as a biocontrol agent.

Bacillus subtilis↗
PubMed2006

Differential responses of germination and seedling establishment in populations of Tragopogon pratensis (Asteraceae).

The establishment phase is an important bottleneck in the life cycle of plants. It consists of two steps that are rarely separated, i.e., the germination of seeds and the establishment of seedlings. Here we report the results of two experiments in which we independently investigated germination and seedling establishment in the greenhouse, under different grass vegetation treatments representing different regeneration niches. Seeds of Tragopogon pratensis from six populations and two habitat types were studied, three from roadside verges and three from hayfields. Germination percentages and germination speed were higher for seeds from roadside verges than for seeds from hayfields, but were little affected by treatment. In contrast, seedling growth was much lower in the tall grass vegetation, than in the short grass and especially the bare soil treatment. Seedling sizes were generally similar for different populations and habitat types. Our results thus show that the two early steps in the establishment phase of plants may respond very differently to the micro-environment, and may have a different selection history. Insight into the ecology and evolution of life histories may require that germination and establishment are considered separately.

Environment↗
PubMed2003

A calmodulin-binding protein from Arabidopsis has an essential role in pollen germination.

Calmodulin (CaM), a ubiquitous multifunctional calcium sensor in all eukaryotes, mediates calcium action by regulating the activity/function of many unrelated proteins. Although calcium and CaM are known to play a crucial role in pollen germination and pollen tube growth, the proteins that mediate their action have not been identified. We isolated three closely related CaM-binding proteins (NPG1, NPGR1, and NPGR2) from Arabidopsis. NPG1 (No Pollen Germination1) is expressed only in pollen, whereas the NPG-related proteins (NPGR1 and NPGR2) are expressed in pollen and other tissues. The bacterially expressed NPG1 bound three isoforms of Arabidopsis CaM in a calcium-dependent manner. To analyze the function of NPG1, we performed a reverse genetics screen and isolated a mutant in which NPG1 is disrupted by a T-DNA insertion. Segregation and molecular analyses of the NPG1 knockout mutant and a cross with a male sterile mutant indicate that the mutated NPG1 is not transmitted through the male gametophyte. Expression of NPG1 in the knockout mutant complemented the mutant phenotype. Analysis of pollen development in the knockout mutant by light microscopy showed normal pollen development. Pollen from NPG1 mutant in the quartet background has confirmed that NPG1 is dispensable for pollen development. However, germination studies with pollen from the mutant in the quartet background indicate that pollen carrying a mutant allele does not germinate. Our genetic, histological, and pollen germination studies with the knockout mutant line indicate that NPG1 is not necessary for microsporogenesis and gametogenesis but is essential for pollen germination.

Alleles↗
PubMed2002

Changes in alpha-and beta-amylase activities during seed germination of African finger millet.

Changes in alpha- and beta-amylase activities in African finger millet (Eleusine coracana (L) Gaertener) were followed during germination. Germination on a small scale was performed at 15 degrees C for 1-10 days and at 20, 25 and 30 degrees C for 1-8 days. alpha- and beta-Amylase activities in malt crude extracts of germinated finger millet were evaluated spectrophotometrically using chromogenic methods. The highest alpha-amylase activity was exhibited in malt flour of finger millet germinated at 15 degrees C for 9 days and at 20 degrees C for 6 days, while the highest beta-amylase activity was displayed in the malt flour germinated for 5 days at 30 degrees C. Thermo-stability of these enzymes in malt extracts was also evaluated. Malt extracts incubated at 40 and 50 degrees C for up to 4 h retained about 84 and 64% of alpha-amylase activities, respectively. There was a substantial decrease in alpha-amylase activity to more than 90% when malt extracts were incubated at 70 and 90 degrees C for 40 and 10 min, respectively. beta-Amylase was completely inactivated when the crude extract was incubated at 70 degrees C for only 10 min. At pH 5.4, alpha-amylase displayed maximum catalytic activity at around 45 degrees C. Optimum temperature for beta-amylase activity at pH 6.0 was between 50 and 55 degrees C. Activity staining for alpha-amylase was also performed and three bands of activity were found in malt extract, each possibly representing an isozyme of alpha-amylase from finger millet.

Africa↗
PubMed1997

Downregulated expression of SHP-1 in Burkitt lymphomas and germinal center B lymphocytes.

We wish to identify developmental changes in germinal center B cells that may contribute to their rapid growth. SHP-1 is an SH2 domain-containing phosphotyrosine phosphatase that negatively regulates activation of B cells and other cells of hematopoietic lineages. We have found that in all 13 EBV-negative and 11 EBV-positive Burkitt lymphomas with a nonlymphoblastoid phenotype, the mean concentration of SHP-1 was reduced to 5% of that of normal B and T cells. The possibility that this diminished expression of SHP-1 was related to the germinal center phenotype of Burkitt lymphomas was supported by the low to absent immunofluorescent staining for SHP-1 in germinal centers, and by the inverse relationship between the concentration of SHP-1 and the expression of the germinal center marker CD38 on purified tonsillar B cells. In CD38-high B cells, SHP-1 concentration was 20% of that of mantle zone B cells from the same donor. This reduction in SHP-1 is comparable to that of cells from motheaten viable mev/mev mice in which there is dysregulated, spontaneous signaling by cytokine and antigen receptors. Therefore, germinal center B cells may have a developmentally regulated, low threshold for cellular activation.

B-Lymphocytes↗
PubMed2005

The effect of gamma irradiation on the germination and growth of certain Nigerian agricultural crops.

Gamma irradiation has been found to be very useful both for sterilisation in medicine and the preservation of food and cereals in nutrition and agriculture. This investigation was carried out to determine the effect of gamma irradiation on the subsequent germination and growth of irradiated seeds. Thirty seeds each of maize, okra and groundnut were irradiated to varying doses of 150, 300, 500, 700, 900, 1000 Gy using the (60)Co gamma cell irradiator facility at the Centre for Energy Research and Development, Obafemi Awolowo University, Ile-Ife. These, as well as the controls (unirradiated seeds), were planted on the same day in an already prepared area of farmland during the rainy season to ensure a constant moisture flow. The times of germination and subsequent growth were monitored. Results show that maize, okra and groundnut seeds needed for planting can be safely stored using gamma irradiation. However, the study reveals that the number of germinated seeds and the growth rate for the crops decrease with increase in the radiation dose the seeds were exposed to. Third-degree polynomial equations were derived which describe the percentage germination of the crops at various levels of exposure. A chart of percentage germination of seeds versus exposure dose is also presented as a quick guide to farmers, policy makers and agricultural institutions.

Crops, Agricultural↗
PubMed2004

Measuring effects of music, noise, and healing energy using a seed germination bioassay.

OBJECTIVE: To measure biologic effects of music, noise, and healing energy without human preferences or placebo effects using seed germination as an objective biomarker. METHODS: A series of five experiments were performed utilizing okra and zucchini seeds germinated in acoustically shielded, thermally insulated, dark, humid growth chambers. Conditions compared were an untreated control, musical sound, pink noise, and healing energy. Healing energy was administered for 15-20 minutes every 12 hours with the intention that the treated seeds would germinate faster than the untreated seeds. The objective marker was the number of seeds sprouted out of groups of 25 seeds counted at 12-hour intervals over a 72-hour growing period. Temperature and relative humidity were monitored every 15 minutes inside the seed germination containers. A total of 14 trials were run testing a total of 4600 seeds. RESULTS: Musical sound had a highly statistically significant effect on the number of seeds sprouted compared to the untreated control over all five experiments for the main condition (p < 0.002) and over time (p < 0.000002). This effect was independent of temperature, seed type, position in room, specific petri dish, and person doing the scoring. Musical sound had a significant effect compared to noise and an untreated control as a function of time (p < 0.03) while there was no significant difference between seeds exposed to noise and an untreated control. Healing energy also had a significant effect compared to an untreated control (main condition, p < 0.0006) and over time (p < 0.0001) with a magnitude of effect comparable to that of musical sound. CONCLUSION: This study suggests that sound vibrations (music and noise) as well as biofields (bioelectromagnetic and healing intention) both directly affect living biologic systems, and that a seed germination bioassay has the sensitivity to enable detection of effects caused by various applied energetic conditions.

Abelmoschus↗
PubMed2004

Effects of germinated brown rice extracts with enhanced levels of GABA on cancer cell proliferation and apoptosis.

In the present work we investigated the effects of brown rice extracts on proliferation and apoptosis of cancer cells. Brown rice extracts were prepared using nongerminated brown rice versus germinated brown rices. Mouse leukemia L1210 cells, human acute lymphoblastic leukemia Molt4 cells, and human cervical cancer HeLa cells were treated with either nongerminated brown rice extract (N ex), water-germinated extract (W ex), chitosan-germinated extract (C ex), glutamic acid-germinated brown rice extract (G ex), or chitosan/glutamic acid-germinated brown rice extract (CG ex). The concentrations of gamma-aminobutyric acid (GABA) in the G ex and CG ex were three and 3.3 times higher than the GABA concentration in the N ex, respectively. The G ex and CG ex retarded significantly the proliferation rates of L1210 and Molt4 cells, and the highest retardation rate was with CG ex. In addition, the G ex and CG ex enhanced significantly apoptosis of the cultured L1210 cells, but no significant apoptosis was seen with the other extracts, which have lower concentrations of GABA than G ex and CG ex. These results show that brown rice extracts with enhanced levels of GABA have an inhibitory action on leukemia cell proliferation and have a stimulatory action on the cancer cell apoptosis.

Animals↗
PubMed2002

Fructokinase and hexokinase from pollen grains of bell pepper (Capsicum annuum L.): possible role in pollen germination under conditions of high temperature and CO2 enrichment.

The processes of pollen grain development and germination depend on the uptake and metabolism of pollen sugars. In pepper (Capsicum annuum L.), initial sugar metabolism includes sucrose hydrolysis by invertase and subsequent phosphorylation of glucose and fructose by hexose kinases. The main objective of this study was to investigate changes in fructokinase (EC 2.7.1.4) and hexokinase (EC.2.7.1.1) activities in pepper flowers during their development, and to study the possible roles of these enzymes in determining pollen germination capacity under high temperature and under CO(2) enrichment, previously shown to modify sugar concentrations in pepper pollen (Aloni et al., 2001 Physiologia Plantarum 112: 505-512). Fructokinase (FK) activity was predominant in pepper pollen, and increased during pollen maturation. Pollen hexokinase (HK) activity was low and did not change throughout pollen development. High-temperature treatment (day/night, 32/26 degrees C) of pepper plants reduced the percentage of pollen that germinated compared with that under normal temperatures (26/22 degrees C), and concomitantly reduced the activity of FK in mature pollen. High temperature also reduced FK and HK activity in the anther. Under high ambient CO(2) (800 micro l l(-1)) pollen FK activity was enhanced. The results suggest that pollen and anther FK may play a role in the regulation of pollen germination, possibly by providing fructose-6-phosphate for glycolysis, or through conversion to UDP-glucose (UDPG) to support the biosynthesis of cell wall material for pollen tube growth. High temperature stress and CO(2) enrichment may influence pollen germination capacity by affecting these pathways.

Calcium Chloride↗