Biomass accumulation and partitioning, photosynthesis, and photosynthetic induction in field-grown maize (Zea mays L.) under low- and high-nitrogen conditions

The objectives of this comparative study were to investigate the responses of biomass accumulation and partitioning to nitrogen supply and to examine the effect of low-nitrogen supply on the photosynthetic responses of maize leaves to steady-state and dynamic light. While the difference in leaf number and stem diameter was not statistically significant, there was a significant difference in plant height between the low-nitrogen and high-nitrogen maize plants. During grain-filling period, the ear leaf of the low-nitrogen maize plants possessed lower values of maximum photosynthetic rate, maximum stomatal conductance, maximum transpiration rate, apparent quantum yield, light compensate point, and carboxylation efficiency than did that of the high-nitrogen maize plants. Contrarily, lower values of intercellular CO₂ concentration and dark respiration rate were observed in the high-nitrogen maize plants. In addition, a slower response to simulated sunflecks was found in the ear leaf of the low-nitrogen maize plants; however, stomatal limitations did not operate in the ear leaf of the high-nitrogen or low-nitrogen maize plants during the photosynthetic induction. As compared to the high-nitrogen maize plants, the low-nitrogen maize plants accumulated much less plant biomass but allocated a greater proportion of biomass to belowground parts. In conclusion, our results suggested that steady-state photosynthetic capacity is restricted by both biochemical and stomatal limitation and the photosynthetic induction is constrained by biochemical limitation alone in low-nitrogen maize plants, and that maize crops respond to low-nitrogen supply in a manner by which more biomass was allocated preferentially to root tissues.     

Purification and functional characterization of the human N-CoR complex: the roles of HDAC3, TBL1 and TBLR1

Corepressors N-CoR and SMRT participate in diverse repression pathways and exist in large protein complexes including HDAC3, TBL1 and TBLR1. However, the roles of these proteins in SMRT-N-CoR complex function are largely unknown. Here we report the purification and functional characterization of the human N-CoR complex. The purified N-CoR complex contains 10-12 associated proteins, including previously identified components and a novel actin-binding protein IR10. We show that TBL1/TBLR1 associates with N-CoR through two independent interactions: the N-terminal region and the C-terminal WD-40 repeats interact with the N-CoR RD1 and RD4 region, respectively. In vitro, TBL1/TBLR1 bind histones H2B and H4, and, importantly, repression by TBL1/TBLR1 correlates with their interaction with histones. By using specific small interference RNAs (siRNAs), we demonstrate that HDAC3 is essential, whereas TBL1 and TBLR1 are functionally redundant but essential for repression by unliganded thyroid hormone receptor. Together, our data reveal the roles of HDAC3 and TBL/TBLR1 and provide evidence for the functional importance of histone interaction in repression mediated by SMRT-N-CoR complexes.     

Single-walled carbon nanotube induction of rat aortic endothelial cell apoptosis: Reactive oxygen species are involved in the mitochondrial pathway

The use of nano-sized materials offers exciting new options in technical and medical applications. Single-walled carbon nanotubes are emerging as technologically important in different industries. However, adverse effects on cells have been reported and this may limit their use. We previously found that 200μg/mL of single-walled carbon nanotubes induce apoptosis in rat aorta endothelial cells. The current study aimed to determine the signaling pathway involved in this process. We found that reactive oxygen species generation was involved in activation of the mitochondria-dependent apoptotic pathway. The finding of apoptosis was supported by a number of morphological and biochemical hallmarks, including chromatin condensation, internucleosomal DNA fragmentation, and caspase-3 activation. In conclusion, our results demonstrate that single-walled carbon nanotubes induce apoptosis in rat aorta endothelial cells and that reactive oxygen species are involved in the mitochondrial pathway.     

Postconditioning attenuates cardiomyocyte apoptosis via inhibition of JNK and p38 mitogen-activated protein kinase signaling pathways

A sequence of intermittent interruptions of oxygen supply (i.e., postconditioning, Postcon) at reoxygenation reduces oxidant-induced cardiomyocyte loss. This study tested the hypothesis that prevention of cardiomyocyte apoptosis by Postcon is mediated by mitogen-activated protein kinases pathways. Primary cultured neonatal rat cardiomyocytes were exposed to 3 h hypoxia followed by 6 h of reoxygenation. Cardiomyocytes were postconditioned by three cycles each of 5 min reoxygenation and 5 min hypoxia after prolonged hypoxia. Relative to hypoxia alone, reoxygenation stimulated expression of JNKs and p38 kinases, corresponding to increased activity of JNKs (phospho-c-Jun) and p38 (phospho-ATF2). The level of TNFα in cell lysates, activity of cytosolic caspases-8, -3, expression of Bax and the number of apoptotic cardiomyocytes were increased while expression of Bcl-2 was decreased with reoxygenation. Consistent with an attenuation in generation of superoxide anions detected by lucigenin-enhanced chemiluminescence at early period of reoxygenation, treatment of cardiomyocytes with Postcon further reduced expression and activity of JNKs and p38 kinases, level of TNFα, the frequency of apoptotic cells and expression of Bax. However, the inhibitory effects of Postcon on these changes were lost when its application was delayed by 5 min after the start of reoxygenation. Addition of a JNK/p38 stimulator, anisomycin into cardiomyocytes at the beginning of reoxygenation eliminated protection by Postcon. These data suggest that 1) hypoxia/reoxygenation elicits cardiomyocyte apoptosis in conjunction with expression and activation of JNK and p38 kinases, release of TNFα, activation of caspases, and an increase in imbalance of pro-/anti-apoptotic proteins; 2) Postcon attenuates cardiomyocyte apoptosis, potentially mediated by inhibiting JNKs/p-38 signaling pathways and reducing TNFα release and caspase expression.     

Possible correlation between high temperature-induced floret sterility and endogenous levels of IAA,GAs and ABA in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

In order to explore the possible physiological mechanism of high temperature induced sterility in rice, we examined the floret sterility and endogenous plant growth regulator contents in pollens of two hybrid rice cultivars Shanyou63 and Teyou559 that are tolerant and susceptible to high temperature, respectively. Indexes of floret sterility, pollen activity, and variation of endogenous indole-3-acetic acid (IAA), gibberellic acids (GAs), abscisic acid (ABA), free proline and soluble proteins in anthers were measured. We found that during the course of high temperature treatment, both cultivars exhibited a marked decrease in pollen activity, pollen germination and floret fertility; however, the high temperature tolerant Shanyou63 showed a much slower rate of decrease than the high temperature susceptible Teyou559. In addition, anthers of both cultivars displayed a decrease in the contents of IAA, GAs, free proline and soluble proteins but an increase in the ABA content. Yet compared to Teyou559, Shanyou63 retained significantly higher levels of free praline and GAs and a lower level of ABA, along with higher pollen vigour and pollen germination rate even after prolonged high temperature treatment. Our study suggests a possible correlation between pollen viability/floret sterility and high temperature-caused changes in IAA, GAs, ABA, free proline and soluble protein contents. The severity in these changes may reflect the variation of rice cultivars in their heat stress sensitivities for floret development.     

Evidence that cardioprotection by postconditioning involves preservation of myocardial opioid content and selective opioid receptor activation

Opioids introduced at reperfusion (R) following ischemia (I) reduce infarct size much like postconditioning, suggesting the hypothesis that postconditioning increases cardiac opioids and activates local opioid receptors. Anesthetized male rats subjected to 30 min regional I and 3 h R were postconditioned with three cycles of 10 s R and 10 s reocclusion at onset of R. Naloxone (NL), its peripherally restricted analog naloxone methiodide, delta-opioid receptor (DOR) antagonist naltrindole (NTI), kappa-opioid receptor antagonist norbinaltorphimine (NorBNI), and mu-opioid receptor (MOR) antagonist H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP) were administered intravenously 5 min before R. The area at risk (AAR) was comparable among groups, and postconditioning reduced infarct size from 57 +/- 2 to 42 +/- 2% (P < 0.05). None of the antagonists alone altered infarct size. All antagonists abrogated postconditioning protection at higher doses. However, blockade of infarct sparing by postconditioning was lost, since tested doses of NL, NTI, NorBNI, and CTAP were lowered. The efficacy of NorBNI declined first at 3.4 micromol/kg, followed sequentially by NTI (1.1), NL (0.37), and CTAP (0.09), suggesting likely MOR and perhaps DOR participation. Representative small, intermediate, and large enkephalins in the AAR were quantified (fmol/mg protein; mean +/- SE). I/R reduced proenkephalin (58 +/- 9 vs. 33 +/- 4; P < 0.05) and sum total of measured enkephalins, including proenkephalin, peptide B, methionine-enkephalin, and methionine-enkephalin-arginine-phenylalanine (139 +/- 17 vs. 104 +/- 7; P < 0.05) compared with shams. Postconditioning increased total enkephalins (89 +/- 8 vs. 135 +/- 5; P < 0.05) largely by increasing proenkephalin (33 +/- 4 vs. 96 +/- 7; P < 0.05). Thus the infarct-sparing effect of postconditioning appeared to involve endogenously activated MORs and possibly DORs, and preservation of enkephalin precursor synthesis in the AAR.     

利用反义RNA技术分析春化相关基因VER17在冬小麦花发育过程中的功能

为了研究小麦春化相关基因VER17的功能,应用反义RNA技术,将VER17基因的反义片段构建到载体pBI121上,通过花粉管通道法获取转基因小麦.对T0代转基因植株GUS染色以及PCR等分子鉴定,得到14株含反义VERJ7基因片段的阳性转基因植株.对T0代和T1代的表型观察结果显示,VER17反义转基因植株开花时间延迟,并且穗的顶部和基部小花出现明显的退化.表明春化相关基因VER17在小麦发育过程中可能起到促进植物开花以及穗顶端和基部花发育的作用,减少小花退化,同时对雄蕊的发育也有影响.     

猪干扰素-γ基因在毕赤酵母中的分泌表达

将去除信号肽的猪干扰素-γ（PoIFN-γ）基因置于酿酒酵母α因子分泌信号的DNA序列后, 构建成pPIC9K-α-PoIFN-γ分泌型重组表达载体, 电转化导入毕赤酵母GS115中,经G418筛选后获得2株多拷贝插入的重组子。SDS-PAGE和Western blot分析结果表明,所获得的重组子能够分泌表达出17kD和23kD左右的PoIFN-γ特异蛋白,其表达量为108mg/L,占培养液总蛋白的60%。实验首次在毕赤酵母表达系统中实现了PoIFN-γ基因的分泌表达。