Effect of various irradiation treatments of plant protoplasts on the transformation rates after direct gene transfer

Summary In P. hybrida and B. nigra an enhancement of transformation rates (direct gene transfer) of about six to seven-fold was obtained after irradiation of protoplasts with 12.5 Gy (X-ray). The effect of protoplast irradiation was similar in experiments where protoplasts were irradiated 1h before transformation (X-ray/DNA) or 1h after completion of the transformation procedure (DNA/X-ray). Increased X-ray doses up to 62.5 Gy resulted in further enhancement of percentages of transformed colonies, indicating a correlation between relative transformation frequencies (RTF) and the doses applied. Estimation of degradation rates of plasmid sequences in plant protoplasts yielded a reduction of plasmid concentration to 50% 8–12 h after transformation. In 1-day-old protoplasts, the level of plasmid fragments dropped to 0%–10% compared to 1h after transformation. The results demonstrate that the integration rates of plasmid sequences into the plant genome may in part be governed by DNA repair mechanisms. This could be an explanation for the observed genotypic dependence of transformation rates in different plant species and plant genotypes. Gene copy number reconstructions revealed enhanced integration rates of plasmid sequences in transformed colonies derived from irradiated protoplasts.     

Characteristics of attention-related body sensations. Temporal stability and associations with measures of body focus,affect, sustained attention,and heart rate variability

Aim: This study investigated the temporal stability and correlates of attention-related body sensations that emerge without external stimulation during rest and due to focused attention on a body part.

Materials: To assess attention-related body sensations, participants were asked to focus on a freely chosen body area with closed eyes, and had to report whether the sensation of that area had changed. Self-report questionnaires were used to assess various aspects of body focus (body awareness, body responsiveness, somatosensory amplification, subjective somatic symptoms), and positive and negative affectivity. Previous experiences in body–mind therapies were also measured. PEBL Continuous Performance Test was used to assess sustained attention. Heart rate variability scores were based on a 3-minute long resting heart rate measurement.

Methods: Fifty-eight university students (22.3?±?3.95 years; 34 females) participated in the study. The stability of attention-related body sensations was measured 8?weeks later on a randomly chosen sub-group (n?=?28).

Results: Attention-related body sensations showed a mediocre temporal stability (r_ρ?=?0.47, p?=?0.012). People reporting attention-related body sensations showed significantly higher body awareness, somatosensory amplification, and resting heart rate; and marginally higher somatic symptoms. No relation was found with body–mind practice, body responsiveness, positive and negative affect, the vagal component of heart rate variability, and performance in the sustained attention task.

Conclusion: Attention-related sensations are relatively stable over time. They are connected to some, but not to all of the aspects of body focus. Further studies are needed to elaborate the influencing stable and situational factors.     

泥沙埋深对苦草和微齿眼子菜及两物种混合分解的影响

为探讨泥沙淤积对水生植物分解的影响,研究了沉水植物苦草(Vallisneria natans)、微齿眼子菜(Pota-mogeton maackianus)及两物种混合在底泥中不同埋深(0 和5 cm)的分解速率和养分动态,实验周期为117d。结果显示:(1)在0和5 cm埋深处理下,苦草、微齿眼子菜及两物种混合的分解速率均表现为苦草最快,微齿眼子菜最慢,物种混合介于两单种之间。与0 cm处理相比,在5 cm埋深处理下苦草、微齿眼子菜及两物种混合的分解速率显著降低(P0.05)。苦草在0和5 cm埋深处理下分解35天后干重剩余率分别为0和43.51%、在5 cm处理下分解82d后干重剩余率为0。微齿眼子菜和两物种混合在5 cm埋深处理下分解117d后的干重剩余率分别提高了31.09%和37.44%。(2)与0 cm处理相比,5 cm埋深处理显著抑制苦草、微齿眼子菜及两物种混合的N、P释放。苦草在0和 5 cm埋深处理下分解35天后N剩余率分别为0和31.28%、P剩余率分别为0和24.45%。在5 cm埋深处理下分解117天后微齿眼子菜N和P剩余率分别提高了19.45%和14.73%、两物种混合N、P剩余率分别提高了41.57%和22.82%。(3)两物种混合在0和5 cm埋深处理下,其分解速率均表现为加和效应,但N、P元素释放在0 cm处理下分别表现为协同效应和加和效应,在5 cm埋深处理下均表现为拮抗效应。(4)随着分解的进行,5 cm埋深处理下的苦草和微齿眼子菜的微生物呼吸速率均显著降低,物种混合的微生物生物量始终低于0 cm处理。这些结果表明泥沙埋深显著降低了苦草、微齿眼子菜及两物种混合的分解速率和N、P元素释放,物种混合的N、P元素释放在分解后期均因沉积作用而产生了拮抗效应。此外,泥沙埋深对分解的抑制作用与微生物呼吸及生物量降低有密切的关系。研究结果可为认识水生植物分解对沉积作用的响应机制提供资料,并为了解水生植物分解对底质营养动态的影响提供参考。     

Sequence-dependent effect of interruptions on microsatellite mutation rate in mismatch repair-deficient human cells

Although microsatellite mutation rates generally increase with increasing length of the repeat tract, interruptions in a microsatellite may stabilize it. We have performed a direct analysis of the effect of microsatellite interruptions on mutation rate and spectrum in cultured mammalian cells. Two mononucleotide sequences (G₁₇ and A₁₇) and a dinucleotide [(CA)₁₇] were compared with interrupted repeats of the same size and with sequences of 8 repeat units. MMR-deficient (MMR⁻) cells were used for these studies to eliminate effects of this repair process. Mutation rates were determined by fluctuation analysis on cells containing a microsatellite sequence at the 5′ end of an antibiotic-resistance gene; the vector carrying this sequence was integrated in the genome of the cells. In general, interrupted sequences had lower mutation rates than perfect ones of the same size, but the magnitude of the difference was dependent upon the sequence of the interrupting base(s). Some interrupted repeats had mutation rates that were lower than those of perfect sequences of the same length but similar to those of half the length. This suggests that interrupting bases effectively divide microsatellites into smaller repeat runs with mutational characteristics different from those of the corresponding full-length microsatellite. We conclude that interruptions decrease microsatellite mutation rate and influence the spectrum of frameshift mutations. The sequence of the interrupting base(s) determines the magnitude of the effect on mutation rate.     

Effects of boundary layer conductance on substomatal pressures of carbon dioxide

Abstract. Gas exchange measurements were made on single leaves of three C₃ and one C₄ species at air speeds of 0.4 and 4.0 m s⁻¹ to determine if boundary layer conductance substantially affected the substomatal pressure of carbon dioxide. Boundary layer conductances to water vapour were 0.4 to 0.5 mol m⁻² s⁻¹ at the lower air speed, and 1.2 to 1.5 mol m⁻² s⁻¹ at the higher air speed. Substomatal carbon dioxide pressures were about 5 Pa lower at low boundary layer conductance in the C₃ species, and about 3 Pa lower in the C₄ species when measurements were made at high and moderate photosynthetic photon flux densities. No evidence of stomatal adjustment to altered boundary layer conductance was found. Photosynthetic rates at high photon flux densities were reduced by about 20% at the low air speed in the C₃ species. The commonly reported values of substomatal carbon dioxide pressure for C₃ and C₄ species were found to occur only when measurements were made at the higher air speed.     

False discovery rate control for multiple testing based on discrete p-values

For multiple testing based on discrete p-values, we propose a false discovery rate (FDR) procedure “BH+” with proven conservativeness. BH+ is at least as powerful as the BH (i.e., Benjamini-Hochberg) procedure when they are applied to superuniform p-values. Further, when applied to mid-p-values, BH+ can be more powerful than it is applied to conventional p-values. An easily verifiable necessary and sufficient condition for this is provided. BH+ is perhaps the first conservative FDR procedure applicable to mid-p-values and to p-values with general distributions. It is applied to multiple testing based on discrete p-values in a methylation study, an HIV study and a clinical safety study, where it makes considerably more discoveries than the BH procedure. In addition, we propose an adaptive version of the BH+ procedure, prove its conservativeness under certain conditions, and provide evidence on its excellent performance via simulation studies.     

辣椒花药培养的胚状体分化及其与成苗率的关系

12个辣椒品种的花药培养的结果表明,所有品种均可诱导出胚状体,其中9个品种可获得健壮的再生植株。每一辣椒品种的适宜植物生长调节利配比不同。不同品种的胚状体诱导率和成苗率有差异。成熟的胚状体均能分化成苗,根先分化或停止发育的胚状体很少成苗。     

ELEVATED CARBON DIOXIDE DIFFERENTIALLY ALTERS THE PHOTOPHYSIOLOGY OF THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE) AND EMILIANIA HUXLEYI (HAPTOPHYTA)1

Increasing anthropogenic carbon dioxide is causing changes to ocean chemistry, which will continue in a predictable manner. Dissolution of additional atmospheric carbon dioxide leads to increased concentrations of dissolved carbon dioxide and bicarbonate and decreased pH in ocean water. The concomitant effects on phytoplankton ecophysiology, leading potentially to changes in community structure, are now a focus of concern. Therefore, we grew the coccolithophore Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler and the diatom strains Thalassiosira pseudonana (Hust.) Hasle et Heimdal CCMP 1014 and T. pseudonana CCMP 1335 under low light in turbidostat photobioreactors bubbled with air containing 390 ppmv or 750 ppmv CO₂. Increased pCO₂ led to increased growth rates in all three strains. In addition, protein levels of RUBISCO increased in the coastal strains of both species, showing a larger capacity for CO₂ assimilation at 750 ppmv CO₂. With increased pCO₂, both T. pseudonana strains displayed an increased susceptibility to PSII photoinactivation and, to compensate, an augmented capacity for PSII repair. Consequently, the cost of maintaining PSII function for the diatoms increased at increased pCO₂. In E. huxleyi, PSII photoinactivation and the counter‐acting repair, while both intrinsically larger than in T. pseudonana, did not change between the current and high‐pCO₂ treatments. The content of the photosynthetic electron transport intermediary cytochrome b6/f complex increased significantly in the diatoms under elevated pCO₂, suggesting changes in electron transport function.