Insights into Hypoxic Systemic Responses Based on Analyses of Transcriptional Regulation in Arabidopsis

We have adopted a hypoxic treatment system in which only roots were under hypoxic conditions. Through analyzing global transcriptional changes in both shoots and roots, we found that systemic signals may be transduced from roots to trigger responses in tissues not directly subjected to hypoxia. The molecular mechanisms of such systemic responses under flooding are currently largely unknown. Using ontological categorization for regulated genes, a systemic managing program of carbohydrate metabolism was observed, providing an example of how systemic responses might facilitate the survival of plants under flooding. Moreover, a proportion of gene expressions that regulated in shoots by flooding was affected in an ethylene signaling mutation, ein2-5. Many systemic-responsive genes involved in the systemic carbohydrate managing program, hormone responses and metabolism, ubiquitin-dependent protein degradation were also affected in ein2-5. These results suggested an important role of ethylene in mediation of hypoxic systemic responses. Genes associated with abscisic acid (ABA) biosynthesis are upregulated in shoots and down regulated in roots. An ABA signaling mutation, abi4-1, affects expression of several systemic responsive genes. These results suggested that regulation of ABA biosynthesis could be required for systemic responses. The implications of these results for the systemic responses of root-flooded Arabidopsis are discussed

A Nanodot Array Modulates Cell Adhesion and Induces an Apoptosis-Like Abnormality in NIH-3T3 Cells

Micro-structures that mimic the extracellular substratum promote cell growth and differentiation, while the cellular reaction to a nanostructure is poorly defined. To evaluate the cellular response to a nanoscaled surface, NIH 3T3 cells were grown on nanodot arrays with dot diameters ranging from 10 to 200 nm. The nanodot arrays were fabricated by AAO processing on TaN-coated wafers. A thin layer of platinum, 5 nm in thickness, was sputtered onto the structure to improve biocompatibility. The cells grew normally on the 10-nm array and on flat surfaces. However, 50-nm, 100-nm, and 200-nm nanodot arrays induced apoptosis-like events. Abnormality was triggered after as few as 24 h of incubation on a 200-nm dot array. For cells grown on the 50-nm array, the abnormality started after 72 h of incubation. The number of filopodia extended from the cell bodies was lower for the abnormal cells. Immunostaining using antibodies against vinculin and actin filament was performed. Both the number of focal adhesions and the amount of cytoskeleton were decreased in cells grown on the 100-nm and 200-nm arrays. Pre-coatings of fibronectin (FN) or type I collagen promoted cellular anchorage and prevented the nanotopography-induced programed cell death. In summary, nanotopography, in the form of nanodot arrays, induced an apoptosis-like abnormality for cultured NIH 3T3 cells. The occurrence of the abnormality was mediated by the formation of focal adhesions

An Intelligent Artificial System : Artificial Immune based Hybrid Genetic Algorithm for the Vehicle Routing Problem

Vehicle routing problems are well-known combinational optimization problems with considerable economic significance. Considering the vehicle routing problem with limited capacity on tree is a problem that often naturally arises in railway, river, and rural road networks. In this paper, we describe an artificial immune system that is distributed, robust, dynamic, diverse and adaptive. It captures many features of the vertebrate immune system and proposed an intelligent artificial system which hybrid genetic and immune algorithm to solve the vehicle routing problem with limited capacity on tree. Computational results show the proposed technique to be very competitive with the best-known heuristic routing procedures providing some new best-known solutions

Restoring Fertility for Novel Interspecific Hybrids between <i>Kalanchoe garambiensis</i> and <i>K. nyikae</i> Using Colchicine Treatment

Interspecific hybridization is an effective strategy in Kalanchoe breeding programs for the introduction of new traits. Wild species within the Kalanchoe genus are valuable genetic resources for providing new horticulture traits and to improve environmental adaptations. However, reproductive barriers associated with fertilization and hybrid sterility must be overcome to produce fertile hybrid progenies. To approach the breeding objectives for Kalanchoe cut flower cultivars with long stem traits and adaptation to tropical/subtropical regions, a tropical species endemic to Taiwan, Kalanchoe garambiensis Kudo, was used as a parent to cross with other long stem Kalanchoe species. Reciprocal crossing was effective in overcoming interspecific unilateral incompatibility in our crossed pairs. One superior hybrid, ‘103-1’, produced capsules without seeds by selfing and backcrossing with pollens from either parent. Other than the seedless trait, failure of pollen releasing from anther, pollen aggregation and no pollen germination in ‘103-1’ suggested its F1 sterility. Colchicine treatments on apical buds of ‘103-1’ successfully overcame potential meiotic abnormalities by doubling ploidy. For the first time, fertile interspecific hybrids of K. garambiensis and K. nyikae Engler were generated. The fertile hybrid has further produced progeny populations by crossing with K. nyikae or K. blossfeldiana von Poelln, ‘Ida’

based Hybrid Genetic Algorithm for the Vehicle Routing Problem

Abstract: Vehicle routing problems are well-known combinational optimization problems with considerable economic significance. Considering the vehicle routing problem with limited capacity on tree is a problem that often naturally arises in railway, river, and rural road networks. In this paper, we describe an artificial immune system that is distributed, robust, dynamic, diverse and adaptive. It captures many features of the vertebrate immune system and proposed an intelligent artificial system which hybrid genetic and immune algorithm to solve the vehicle routing problem with limited capacity on tree. Computational results show the proposed technique to be very competitive with the best-known heuristic routing procedures providing some new best-known solutions

Arabidopsis Hsa32, a Novel Heat Shock Protein, Is Essential for Acquired Thermotolerance during Long Recovery after Acclimation

Plants and animals share similar mechanisms in the heat shock (HS) response, such as synthesis of the conserved HS proteins (Hsps). However, because plants are confined to a growing environment, in general they require unique features to cope with heat stress. Here, we report on the analysis of the function of a novel Hsp, heat-stress-associated 32-kD protein (Hsa32), which is highly conserved in land plants but absent in most other organisms. The gene responds to HS at the transcriptional level in moss (Physcomitrella patens), Arabidopsis (Arabidopsis thaliana), and rice (Oryza sativa). Like other Hsps, Hsa32 protein accumulates greatly in Arabidopsis seedlings after HS treatment. Disruption of Hsa32 by T-DNA insertion does not affect growth and development under normal conditions. However, the acquired thermotolerance in the knockout line was compromised following a long recovery period (>24 h) after acclimation HS treatment, when a severe HS challenge killed the mutant but not the wild-type plants, but no significant difference was observed if they were challenged within a short recovery period. Quantitative hypocotyl elongation assay also revealed that thermotolerance decayed faster in the absence of Hsa32 after a long recovery. Similar results were obtained in Arabidopsis transgenic plants with Hsa32 expression suppressed by RNA interference. Microarray analysis of the knockout mutant indicates that only the expression of Hsa32 was significantly altered in HS response. Taken together, our results suggest that Hsa32 is required not for induction but rather maintenance of acquired thermotolerance, a feature that could be important to plants

The AP2/ERF Transcription Factor AtERF73/HRE1 Modulates Ethylene Responses during Hypoxia in Arabidopsis1[W][OA]

A number of APETALA2 (AP2)/ETHYLENE RESPONSE FACTOR (ERF) genes have been shown to function in abiotic and biotic stress responses, and these genes are often induced by multiple stresses. We report here the characterization of an AP2/ERF gene in Arabidopsis (Arabidopsis thaliana) that is specifically induced during hypoxia. We show that under normoxic conditions, the expression of AtERF73/HRE1 can be induced by exogenous addition of 1-aminocyclopropane-1-carboxylic acid and that a combination of hypoxia and 1-aminocyclopropane-1-carboxylic acid results in hyperinduction of AtERF73/HRE1 expression. In addition, hypoxic induction of AtERF73/HRE1 is reduced but not completely abolished in ethylene-insensitive mutants and in the presence of inhibitors of ethylene biosynthesis and responses. These results suggest that, in addition to ethylene, an ethylene-independent signal is also required to mediate hypoxic induction of AtERF73/HRE1. To assess the role of AtERF73/HRE1, we generated three independent RNA interference (RNAi) knockdown lines of AtERF73/HRE1. Under normoxic conditions, the AtERF73/HRE1-RNAi seedlings displayed increased ethylene sensitivity and exaggerated triple responses, indicating that AtERF73/HRE1 might play a negative regulatory role in modulating ethylene responses. Gas chromatography analyses showed that the production of ethylene was similar between wild-type and RNAi lines under hypoxia. Quantitative reverse transcription-polymerase chain reaction analyses showed that hypoxia-inducible genes could be affected by AtERF73/HRE1-RNAi lines in two different ways: hypoxic induction of glycolytic and fermentative genes was reduced, whereas induction of a number of peroxidase and cytochrome P450 genes was increased. Taken together, our results show that AtERF73/HRE1 is involved in modulating ethylene responses under both normoxia and hypoxia