Effects of clinorotation and microgravity on sweet clover columella cells treated with cytochalasin D

The cytoskeleton of columella cells is believed to be involved in maintaining the developmental polarity of cells observed as a reproducible positioning of cellular organelles. It is also implicated in the transduction of gravitropic signals. Roots of sweet clover ( Melilotus alba L.) seedlings were treated with a microfilament disrupter, cytochalasin D, on a slowly rotating horizontal clinostat (2 rpm). Electron micrographs of treated columella cells revealed several ultrastructural effects including repositioning of the nucleus and the amyloplasts and the formation of endoplasmic reticulum (ER) whorls. However, experiments performed during fast clinorotation (55 rpm) showed an accumulation (but no whorling) of a disorganized ER network at the proximal and distal pole and a random distribution of the amyloplasts. Therefore, formation of whorls depends upon the speed of clinorotation, and the overall impact of cytochalasin D suggests the necessity of microfilaments in organelle positioning. Interestingly, a similar drug treatment performed in microgravity aboard the US Space Shuttle Endeavour (STS-54, January 1993) caused a displacement of ER membranes and amyloplasts away from the distal plasma membrane. In the present study, we discuss the role of microfilaments in maintaining columella cell polarity and the utility of clinostats to simulate microgravity.     

Cortical microtubules in sweet clover columella cells developed in microgravity

Electron micrographs of columella cells from sweet clover seedlings grown and fixed in microgravity revealed longitudinal and cross sectioned cortical microtubules. This is the first report demonstrating the presence and stability of this network in plants in microgravity.     

Mode of high temperature injury to wheat during grain development

High temperature stress adversely affects wheat growth in many important production regions, but the mode of injury is unclear. Wheat ( Triticum aestivum L. cv. Newton) was grown under controlled conditions to determine the relative magnitude and sequences of responses of source and sink processes to high temperature stress during grain development. Regimes of 25°C day/15°C night, 30°C day/20°C night, and 35°C day/25°C night from 5 days after anthesis to maturity differentially affected source and sink processes. High temperatures accelerated the normal decline in viable leaf blade area and photosynthetic activities per unit leaf area. Electron transport, as measured by Hill reaction activity, declined earlier and faster than other photosynthetic processes at the optimum temperature of 25/15 °C and at elevated temperatures. Changes in RUBP carboxylase activities were similar in direction but smaller in magnitude than changes in photosynthesic rate. Increased protease activity during senscence was markedly accentuated by high temperature stress. Specific protease activity increased 4-fold at 25/15 °C and 28-fold at 35/25 °C from 0 to 21 days after initiation of temperature treatments. Grain-filling rate decreased from the lowest to the highest temperature, but the change was smaller than the decrease in grain-filling duration at the same temperatures. We concluded that a major effect of high temperature is acceleration of senescence, including cessation of vegetative and reproductive growth, deterioration of photosynthetic activities, and degradation of proteinaceous constituents.     

The influence of volume of exercise on early adaptations to strength training

The purpose of this investigation was to compare the effects of single-set strength training and 3-set strength training during the early phase of adaptation in 18 untrained male subjects (age, 20-30 years). After initial testing, subjects were randomly assigned to either the 3L-1U group (n = 8), which trained 3 sets in leg exercises and 1 set in upper-body exercises, or the 1L-3U group (n = 10), which trained 1 set in leg exercises and 3 sets in upper-body exercises. Testing was conducted at the beginning and at the end of the study and consisted of 2 maximal isometric tests (knee extension and bench press) and 6 maximal dynamic tests (1 repetition maximum [1RM] tests). Subjects trained 3 days per week for 6 weeks. After warm-up, subjects performed 3 leg exercises and 4 upper-body exercises. In both groups, each set consisted of 7 repetitions (reps) with the load supposed to induce muscular failure after the seventh rep (7RM load). After 6 weeks of training, 1RM performance in all training exercises was significantly increased (10-26%, p < 0.01) in both groups. The relative increase in 1RM load in the 3 leg exercises was significantly greater in the 3L-1U group than in the 1L-3U group (21% vs. 14%, p = 0.01). However, the relative increase in 1RM load in the 3 upper-body exercises was similar in the 3L-1U group (16%) and the 1L-3U group (14%). These results show a superior adaptation to 3-set strength training, compared with 1-set strength training, in leg exercises but not in upper-body exercises during the early phase of adaptation.     

Water-soluble chlorophyll protein (WSCP) of Arabidopsis is expressed in the gynoecium and developing silique

Water-soluble chlorophyll protein (WSCP) has been found in many Brassicaceae, most often in leaves. In many cases, its expression is stress-induced, therefore, it is thought to be involved in some stress response. In this work, recombinant WSCP from Arabidopsis thaliana (AtWSCP) is found to form chlorophyll-protein complexes in vitro that share many properties with recombinant or native WSCP from Brassica oleracea, BoWSCP, including an unusual heat resistance up to 100°C in aqueous solution. A polyclonal antibody raised against the recombinant apoprotein is used to identify plant tissues expressing AtWSCP. The only plant organs containing significant amounts of AtWSCP are the gynoecium in open flowers and the septum of developing siliques, specifically the transmission tract. In fully grown but still green siliques, the protein has almost disappeared. Possible implications for AtWSCP functions are discussed.     

Hypoxia inducible factor activates the transforming growth factor-alpha/epidermal growth factor receptor growth stimulatory pathway in VHL(-/-) renal cell carcinoma cells

Bi-allelic-inactivating mutations of the VHL tumor suppressor gene are found in the majority of clear cell renal cell carcinomas (VHL(-/-) RCC). VHL(-/-) RCC cells overproduce hypoxia-inducible genes as a consequence of constitutive, oxygen-independent activation of hypoxia inducible factor (HIF). While HIF activation explains the highly vascularized nature of VHL loss lesions, the relative role of HIF in oncogenesis and loss of growth control remains unknown. Here, we report that HIF plays a central role in promoting unregulated growth of VHL(-/-) RCC cells by activating the transforming growth factor-alpha (TGF-alpha)/epidermal growth factor receptor (EGF-R) pathway. Dominant-negative HIF and enzymatic inhibition of EGF-R were equally efficient at abolishing EGF-R activation and serum-independent growth of VHL(-/-) RCC cells. TGF-alpha is the only known EGF-R ligand that has a VHL-dependent expression profile and its overexpression by VHL(-/-) RCC cells is a direct consequence of HIF activation. In contrast to TGF-alpha, other HIF targets, including vascular endothelial growth factor (VEGF), were unable to stimulate serum-independent growth of VHL(-/-) RCC cells. VHL(-/-) RCC cells expressing reintroduced type 2C mutants of VHL, and which retain the ability to degrade HIF, fail to overproduce TGF-alpha and proliferate in serum-free media. These data link HIF with the overproduction of a bona fide renal cell mitogen leading to activation of a pathway involved in growth of renal cancer cells. Moreover, our results suggest that HIF might be involved in oncogenesis to a much higher extent than previously appreciated.     

Markers associated with a QTL for grain yield in wheat under drought

Drought is a major abiotic stress that adversely affects wheat production in many regions of the world. The objective of this study was to identify quantitative trait loci (QTL) controlling grain yield and yield components under reduced moisture. A cross between common wheat cultivars ‘Dharwar Dry’ (drought tolerant) and ‘Sitta’ was the source of one hundred twenty-seven recombinant inbred lines evaluated for two-seasons in a field under differing soil moisture regimes in Ciudad Obregon, Sonora, Mexico. An SSR/EST-STS marker map was constructed and a grain yield QTL on the proximal region of chromosome 4AL was found to have a significant impact on performance under reduced moisture. This region was associated with QTL for grain yield, grain fill rate, spike density, grains m⁻², biomass production, biomass production rate, and drought susceptibility index (DSI). Molecular markers associated with these traits explained 20, 33, 15, 23, 30, 26, and 41% of phenotypic variation, respectively on chromosome 4A. Microsatellite locus Xwmc89 was associated with all significant QTL covering a 7.7 centiMorgans (cM) region and generally explained the greatest proportion of phenotypic variation. The alleles associated with enhanced performance under drought stress were contributed by Dharwar Dry. Microsatellite marker wmc89 may be useful for marker assisted selection to enhance drought tolerance.