A SEPALLATA gene is involved in the development and ripening of strawberry (Fragaria X ananassa Duch.) fruit, a non-climacteric tissue

Climacteric and non-climacteric fruits have traditionally been viewed as representing two distinct programmes of ripening associated with differential respiration and ethylene hormone effects. In climacteric fruits, such as tomato and banana, the ripening process is marked by increased respiration and is induced and co-ordinated by ethylene, while in non-climacteric fruits, such as strawberry and grape, it is controlled by an ethylene-independent process with little change in respiration rate. The two contrasting mechanisms, however, both lead to texture, colour, and flavour changes that probably reflect some common programmes of regulatory control. It has been shown that a SEPALLATA(SEP)4-like gene is necessary for normal ripening in tomato. It has been demonstrated here that silencing a fruit-related SEP1/2-like (FaMADS9) gene in strawberry leads to the inhibition of normal development and ripening in the petal, achene, and receptacle tissues. In addition, analysis of transcriptome profiles reveals pleiotropic effects of FaMADS9 on fruit development and ripening-related gene expression. It is concluded that SEP genes play a central role in the developmental regulation of ripening in both climacteric and non-climacteric fruits. These findings provide important information to extend the molecular control of ripening in a non-climacteric fruit beyond the limited genetic and cultural options currently available.

Nutrients distribution in diseased coffee leaf tissue

Knowing the structure and distribution of nutrients in plant tissues can clarify some mechanisms of pathogen attack in plants and plant defense against infection, thus helping management strategies. The aim of this study was verify differences in distribution of mineral nutrients in coffee leaf tissues around foliar lesions of bacterial blight of coffee, blister spot, cercospora leaf, phoma leaf spot and coffee leaf rust. Fragments of leaf tissue surrounding the lesions were dehydrated in silica gel, carbon covered and subjected to X-ray microanalysis (MAX). Thirty-three chemical elements were detected in leaf tissue; however, there was variation in potassium and calcium contents surrounding the lesions. The highest potassium content was found in asymptomatic tissues surrounding the lesions, decreasing toward the transition zone and reaching minimum content in symptomatic tissues. The highest calcium content was found in symptomatic tissues, decreasing toward the transition zone and reaching minimum content in asymptomatic tissues. Therefore, MAX can be used to analyze the composition and distribution of nutrients in plant tissues and, if associated with mineral nutrition, it may help understand host-pathogen relationships and plant disease management.

Shared Escovopsis parasites between leaf-cutting and non-leaf-cutting ants in the higher attine fungus-growing ant symbiosis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

500 ml of blood loss does not decrease non-invasive tissue oxygen saturation (StO2) as measured by near infrared spectroscopy - A hypothesis generating pilot study in healthy adult women

Background The goal when resuscitating trauma patients is to achieve adequate tissue perfusion. One parameter of tissue perfusion is tissue oxygen saturation (StO2), as measured by near infrared spectroscopy. Using a commercially available device, we investigated whether clinically relevant blood loss of 500 ml in healthy volunteers can be detected by changes in StO2 after a standardized ischemic event. Methods We performed occlusion of the brachial artery for 3 minutes in 20 healthy female blood donors before and after blood donation. StO2 and total oxygenated tissue hemoglobin (O2Hb) were measured continuously at the thenar eminence. 10 healthy volunteers were assessed in the same way, to examine whether repeated vascular occlusion without blood donation exhibits time dependent effects. Results Blood donation caused a substantial decrease in systolic blood pressure, but did not affect resting StO2 and O2Hb values. No changes were measured in the blood donor group in the reaction to the vascular occlusion test, but in the control group there was an increase in the O2Hb rate of recovery during the reperfusion phase. Conclusion StO2 measured at the thenar eminence seems to be insensitive to blood loss of 500 ml in this setting. Probably blood loss greater than this might lead to detectable changes guiding the treating physician. The exact cut off for detectable changes and the time effect on repeated vascular occlusion tests should be explored further. Until now no such data exist.

Detection of Ca2+-Dependent Transglutaminase Activity in Root and Leaf Tissue of Monocotyledonous and Dicotyledonous Plants

Protein extracted from root and leaf tissue of the dicotyledonous plants pea (Pisum sativum) and broad bean (Vicia faba) and the monocotyledonous plants wheat (Triticum aestivum) and barley (Hordeum vulgare) were shown to catalyze the incorporation of biotin-labeled cadaverine into microtiter-plate-bound N′,N′-dimethylcasein and the cross-linking of biotin-labeled casein to microtiter-plate-bound casein in a Ca2+-dependent manner. The cross-linking of biotinylated casein and the incorporation of biotin-labeled cadaverine into N′,N′-dimethylcasein were time-dependent reactions with a pH optimum of 7.9. Transglutaminase activity was shown to increase over a 2-week growth period in both the roots and leaves of pea. The product of transglutaminase's protein-cross-linking activity, ε-(γ-glutamyl)-lysine isodipeptide, was detected in root and shoot protein from pea, broad bean, wheat, and barley by cation-exchange chromatography. The presence of the isodipeptide was confirmed by reversed-phase chromatography. Hydrolysis of the isodipeptide after cation-exchange chromatography confirmed the presence of glutamate and lysine.

Non root feeding with two sources of zinc on Coffea arábica L. 'mundo novo'(B.Rodr.) Choussy

A trial was carried out on an eight old coffee plantation with visible zinc problems. The plantation was situated nearly the city of Jaú (22º30'S, 48º30'W). State of São Paulo, Brazil. The soil is classified as medium texture Oxisol of low base saturation (Latossol Vermelho Amarelo - fase arenosa). The pulverization program started in november 1977, followed in march and July 1978 (heavy harvest) and ended in march and July 1979 (light harvest). Is should be mentioned that a well reconized characteristic of arábica coffe is its habit of biennial bearing, a very heavy harvest is most often followed by a light load the next year. The following treatments and amounts of chemicals per cova hole (4 trees) were tested in accordance with a random block design: 1. 1 g of zinc (zinc sulphate, 0.5%) 2. 3 g of nitrogen (urea, 1.3%) 3. 1 g of zinc + 3 g of nitrogen (zinc sulphate 0.5% + urea 1.3%) 4. 0.25 g, 0.50 g, 1.00 g, 2.00 g of zinc plus 0.75 g, 1.50 g, 3.00 g and 6.00 of nitrogen (correspondent to NZN* 15-0-0-5 as 0.75%, 1-5%, 3.0% and 6.0% by v/v). Foliar absorption data were obtained by collecting the 3rd and 4th pairs of the coffee leaves and analysed them for N, P, K, Ca, Mg, S, B, Cu, Fe, Mn, and Zn. The main results may be summarized as follows: 1. The maximum calculated yields of clean coffee were obtained by the applications of 5.84 1 of NZN (1.13%) per hectare. 2. The applications of zinc sulphate (0.5%) and urea (1.3%) together or separate did not affected the coffee bean production. 3. The applications of 15.0 1 of NZN per hectare reduced the coffee yields. 4. Leaf damages and burning symptoms were observed by the applications of urea (1.3%) plus zinc sulphate (0.5%) and larger doses than 7.5 1 of NZN per hectare. 5. Leaf tissue analysis show that the concentrations of the elements were affecred by the age of the leaves and by the yields of the coffee trees. 6. The applications of increasing doses of NZN causes an increase in the concentration of zinc, manganese and boron in the leaves and decreased the concentration in calcium and potassium the leaves. 7. The concentration of zinc in the leaves associated with the heavy harvest, in July, was 70.0 ppm.

Chloroacetamid Spray Drift and Leaf Tatters in Hackberry

During the last decade, leaf tatters has been reported in white oak and hackberry across several Midwestern states. Herbicide spray drift studies have shown that chloroacetamides can induce leaf tatters. The objectives of this research were to: 1) identify vulnerable bud developmental stages in hackberry and 2) determine if different commercial chloroacetamides affect severity of leaf tatters. In 2008, a preliminary spray drift experiment was conducted on mature trees from a former hackberry provenance test stand. Acetochlor (Harness), S-metolachlor (Dual II Magnum), and dimethenamid (Outlook) were applied at concentrations approximating 27%, 54%, 81%, or 108% of the recommended field rate. Three developmental stages before bud burst were present on the selected trees. Leaf tatters did not develop on the selected hackberry trees. However, symptoms were observed on neighboring, non-target hackberry trees, which had been in the leaf unfolding and expanding stages at the time of spraying. In 2009, three year old hackberry seedlings were treated with 1%, 10%, and 100% of the recommended field rate of acetochlor (Harness), S-metolachlor (Dual II Magnum), and dimethenamid (Outlook). Folded buds and two unfolding leaf developmental stages were present on seedlings. Another spray study was conducted on 32 mature hackberry trees from the provenance stand. A solution of 5608 mg a.i./L dimethenamid (Outlook) was applied to trees in the unfolding and/or expanding leaf stage. Treated trees represented four provenances. Image analysis was used to calculate seedling and mature tree leaf areas and estimate the seedling percentage of leaf tissue loss. Foliar damage was not significantly different between seedlings treated with water, 1%, or 10% of the field rate. Foliar damage was significantly different between seedlings treated with 1% or 100% of the field rate, and between seedlings treated with 10% or 100% of the field rate. Foliar damage in seedlings was not significantly different between the developmental stages. Additionally, symptoms of leaf tatters were observed on the treated mature hackberry. Future studies should focus on chloroacetamide concentrations above 10% of the recommended field rate.

Transforming growth factor alpha immunoreactivity. A study in hepatocellular carcinoma and in non-neoplastic liver tissue

Background. Transforming growth factor alpha (TGF alpha) is an important mitogen that binds to epidermal growth factor receptor and is associated with the development of several tumors. Aims. Assessment of the immunoexpression of TGF alpha in hepatocellular carcinoma (HCC) and in non-neoplastic liver tissue and its relationship to morphological patterns of HCC. Material and methods. The immunohistochemical expression of TGF alpha was studied in 47 cases of HCC (27 multinodular, 20 nodular lesions). Five lesions measured up to 5 cm and 15 lesions above 5 cm. Thirty-two cases were graded as I or II and 15 as III or IV. The non-neoplastic tissue was examined in 40 cases, of which 22 had cirrhosis. HBsAg and anti-HCV were positive in 5/38 and 15/37 patients, respectively. The statistical analysis for possible association of immunostaining of TGF alpha and pathological features was performed through chi-square test. Results. TGF alpha was detected in 31.9% of the HCC and in 42.5% of the non-neoplastic. There was a statistically significant association between the expression of TGF alpha and cirrhosis (OR = 8.75, 95% CI = [1.93, 39.75]). The TGF alpha was detected more frequently in patients anti-HCV(+) than in those HBsAg(+). The immunoexpression of TGF alpha was not found related to tumor size or differentiation. In conclusion the TGF alpha is present in hepatocarcinogenesis in HBV negative patients. Further analysis is needed to examine the involvement of TGF alpha in the carcinogenesis associated with HCV and other possible agents. In addition, TGF alpha has an higher expression in hepatocyte regeneration and proliferation in cirrhotic livers than in HCC.

Correlation between MMPs and their inhibitors in breast cancer tumor tissue specimens and in cell lines with different metastatic potential

Background: The metastatic disease rather than the primary tumor itself is responsible for death in most solid tumors, including breast cancer. The role of matrix metalloproteinases ( MMPs), tissue inhibitors of MMPs (TIMPs) and Reversion-inducing cysteine-rich protein with Kazal motifs ( RECK) in the metastatic process has previously been established. However, in all published studies only a limited number of MMPs/MMP inhibitors was analyzed in a limited number of cell lines. Here, we propose a more comprehensive approach by analyzing the expression levels of several MMPs (MMP-2, MMP-9 and MMP-14) and MMP inhibitors (TIMP-1, TIMP-2 and RECK) in different models ( five human breast cancer cell lines, 72 primary breast tumors and 30 adjacent normal tissues). Methods: We analyzed the expression levels of MMP-2, MMP-9 and MMP-14 and their inhibitors (TIMP-1, TIMP-2 and RECK) by quantitative RT-PCR (qRT-PCR) in five human breast cancer cell lines presenting increased invasiveness and metastatic potential, 72 primary breast tumors and 30 adjacent normal tissues. Moreover, the role of cell-extracellular matrix elements interactions in the regulation of expression and activity of MMPs and their inhibitors was analyzed by culturing these cell lines on plastic or on artificial ECM (Matrigel). Results: The results demonstrated that MMPs mRNA expression levels displayed a positive and statistically significant correlation with the transcriptional expression levels of their inhibitors both in the cell line models and in the tumor tissue samples. Furthermore, the expression of all MMP inhibitors was modulated by cell-Matrigel contact only in highly invasive and metastatic cell lines. The enzyme/inhibitor balance at the transcriptional level significantly favors the enzyme which is more evident in tumor than in adjacent non-tumor tissue samples. Conclusion: Our results suggest that the expression of MMPs and their inhibitors, at least at the transcriptional level, might be regulated by common factors and signaling pathways. Therefore, the multi-factorial analysis of these molecules could provide new and independent prognostic information contributing to the determination of more adequate therapy strategies for each patient.`

Intercellular transport of epidermis-expressed MADS domain transcription factors and their effect on plant morphology and floral transition

P>During the lifetime of an angiosperm plant various important processes such as floral transition, specification of floral organ identity and floral determinacy, are controlled by members of the MADS domain transcription factor family. To investigate the possible non-cell-autonomous function of MADS domain proteins, we expressed GFP-tagged clones of AGAMOUS (AG), APETALA3 (AP3), PISTILLATA (PI) and SEPALLATA3 (SEP3) under the control of the MERISTEMLAYER1 promoter in Arabidopsis thaliana plants. Morphological analyses revealed that epidermal overexpression was sufficient for homeotic changes in floral organs, but that it did not result in early flowering or terminal flower phenotypes that are associated with constitutive overexpression of these proteins. Localisations of the tagged proteins in these plants were analysed with confocal laser scanning microscopy in leaf tissue, inflorescence meristems and floral meristems. We demonstrated that only AG is able to move via secondary plasmodesmata from the epidermal cell layer to the subepidermal cell layer in the floral meristem and to a lesser extent in the inflorescence meristem. To study the homeotic effects in more detail, the capacity of trafficking AG to complement the ag mutant phenotype was compared with the capacity of the non-inwards-moving AP3 protein to complement the ap3 mutant phenotype. While epidermal expression of AG gave full complementation, AP3 appeared not to be able to drive all homeotic functions from the epidermis, perhaps reflecting the difference in mobility of these proteins.

On the extent of genetic variation for transpiration efficiency in sorghum

A glasshouse study examined 49 diverse sorghum lines for variation in transpiration efficiency. Three of the 49 lines grown were Sorghum spp, native to Australia; one was the major weed Johnson grass (Sorghum halepense), and the remaining 45 lines were cultivars of Sorghum bicolor. All plants were grown under non-limiting water and nutrient conditions using a semi-automatic pot watering system designed to facilitate accurate measurement of water use. Plants were harvested 56-58 days after sowing and dry weights of plant parts were determined. Transpiration efficiency differed significantly among cultivars. The 3 Australian native sorghums had much lower transpiration efficiency than the other 46 cultivars, which ranged from 7.7 to 6.0 g/kg. For the 46 diverse cultivars, the ratio of range in transpiration efficiency to its l.s.d. was 2.0, which was similar to that found among more adapted cultivars in a previous study. This is a significant finding as it suggests that there is likely to be little pay-off from pursuing screening of unadapted material for increased variation in transpiration efficiency. It is necessary, however, also to examine absolute levels of transpiration efficiency to determine whether increased levels have been found. The cultivar with greatest transpiration efficiency in this study (IS9710) had a value 9% greater (P < 0.05) than the accepted standard for adapted sorghum cultivars. The potential impact of such an increase in transpiration efficiency warrants continued effort to capture it. Transpiration efficiency has been related theoretically and experimentally to the degree of carbon isotope discrimination in leaf tissue in sorghum, which thus offers a relatively simple selection index. In this study, the variation in transpiration efficiency was not related simply to carbon isotope discrimination. Significant associations of transpiration efficiency with ash content and indices of photosynthetic capacity were found. However, the associations were not strong. These results suggest that a simple screening technique could not be based on any of the measures or indices analysed in this study. A better understanding of the physiological basis of the observed genetic differences in transpiration efficiency may assist in developing reliable selection indices. It was concluded that the potential value of the improvement in transpiration efficiency over the accepted standard and the degree of genetic variation found warrant further study on this subject. It was suggested that screening for genetic variation under water-limiting conditions may provide useful insights and should be pursued.

In vitro growth and leaf anatomy of Cattleya walkeriana (Gardner, 1839) grown in natural ventilation system

Natural ventilation system facilitates gaseous exchanges in in vitro plants promoting changes in the leaf tissue, which can be evaluated through the leaf anatomy, and it allows a cultivation closer to the photoautrophic micropropagation. The objective of this work was to evaluate the effects on in vitro growth and on the leaf anatomy of Cattleya walkeriana grown in natural and conventional ventilation system with different concentrations of sucrose (0; 15; 30 and 45 L-1) combined with different cultivation systems (conventional micropropagation and natural ventilation system). The culture medium was composed of MS salts, solidified with 7 g L-1 of agar and pH adjusted to 5.8. Forty milliliters of culture medium were distributed in 250 mL flasks, autoclaved at 120 ºC for 20 minutes. The greater plant growth, as well as the greater thickness of the mesophyll was observed with the use of 20 g L-1 sucrose in natural ventilation system. Plants grown in natural ventilation system showed a thicker leaf mesophyll, which is directly related to photoautotrophic crops. The natural ventilation system induced more elliptical stomata and probably more functional formats.

Tissue-adapted invasion strategies of the rice blast fungus Magnaporthe oryzae.

Magnaporthe oryzae causes rice blast, the most serious foliar fungal disease of cultivated rice (Oryza sativa). During hemibiotrophic leaf infection, the pathogen simultaneously combines biotrophic and necrotrophic growth. Here, we provide cytological and molecular evidence that, in contrast to leaf tissue infection, the fungus adopts a uniquely biotrophic infection strategy in roots for a prolonged period and spreads without causing a loss of host cell viability. Consistent with a biotrophic lifestyle, intracellularly growing hyphae of M. oryzae are surrounded by a plant-derived membrane. Global, temporal gene expression analysis used to monitor rice responses to progressive root infection revealed a rapid but transient induction of basal defense-related gene transcripts, indicating perception of the pathogen by the rice root. Early defense gene induction was followed by suppression at the onset of intracellular fungal growth, consistent with the biotrophic nature of root invasion. By contrast, during foliar infection, the vast majority of these transcripts continued to accumulate or increased in abundance. Furthermore, induction of necrotrophy-associated genes during early tissue penetration, previously observed in infected leaves, was not seen in roots. Collectively, our results not only report a global characterization of transcriptional root responses to a biotrophic fungal pathogen but also provide initial evidence for tissue-adapted fungal infection strategies.

Role of major histocompatibility complex class II expression by non-hematopoietic cells in autoimmune and inflammatory disorders: facts and fiction.

It is well established that interactions between CD4(+) T cells and major histocompatibility complex class II (MHCII) positive antigen-presenting cells (APCs) of hematopoietic origin play key roles in both the maintenance of tolerance and the initiation and development of autoimmune and inflammatory disorders. In sharp contrast, despite nearly three decades of intensive research, the functional relevance of MHCII expression by non-hematopoietic tissue-resident cells has remained obscure. The widespread assumption that MHCII expression by non-hematopoietic APCs has an impact on autoimmune and inflammatory diseases has in most instances neither been confirmed nor excluded by indisputable in vivo data. Here we review and put into perspective conflicting in vitro and in vivo results on the putative impact of MHCII expression by non-hematopoietic APCs-in both target organs and secondary lymphoid tissues-on the initiation and development of representative autoimmune and inflammatory disorders. Emphasis will be placed on the lacunar status of our knowledge in this field. We also discuss new mouse models-developed on the basis of our understanding of the molecular mechanisms that regulate MHCII expression-that constitute valuable tools for filling the severe gaps in our knowledge on the functions of non-hematopoietic APCs in inflammatory conditions.

Acclimatization and leaf anatomy of micropropagated fig plantlets

The survival of micropropagated plants during and after acclimatization is a limiting process to plant establishment. There is little information on how the anatomy of vegetative organs of Ficus carica can be affected by culture conditions and acclimatization. The present research aimed to study the effects of time on culture medium and substrates during the acclimatization of fig tree plantlets produced in vitro, characterizing some leaf anatomy aspects of plantlets cultured in vitro and of fig trees produced in field. Plantlets previously multiplied in vitro were separated and transferred into Wood Plant Medium (WPM) where they were kept for 0, 15, 30, 45 and 60 days. Different substrates were tested and studies on leaf anatomy were performed in order to compare among plantlets grown in vitro, plantlets under 20, 40 and 60 days of acclimatization, and field grown plants. Keeping plantlets for 30 days in WPM allowed better development in Plantmax during acclimatization. Field grown plants presented higher number of stomata, greater epicuticular wax thickness and greater leaf tissue production compared to in vitro ones. The leaf tissues of in vitro plantlets show little differentiation and have great stomata number compared with acclimatized plants, which reduce the number of stomata during the acclimatization process.