Digital imaging approaches for phenotyping whole plant nitrogen and phosphorus response in Bracbypodium distacbyon

This work evaluates the phenotypic response of the model grass （Brachypodium distacbyon （L.） P. Beauv.） to nitrogen and phosphorus nutrition using a combination of imaging techniques and destructive harvest of shoots and roots. Reference line Bd21-3 was grown in pots using 11 phosphorus and 11 nitrogen concentrations to establish a dose-response curve. Shoot biovolume and biomass, root length and biomass, and tissue phosphorus and nitrogen concentrations increased with nutrient concentration. Shoot biovolume, estimated by imaging, was highly correlated with dry weight （R2 〉 0.92） and both biovolume and growth rate responded strongly to nutrient availability. Higher nutrient supply increased nodal root length more than other root types. Photochemical efficiency was strongly reduced by low phosphorus concentrations as early as 1 week after germination, suggesting that this measurement may be suitable for high throughput screening of phosphorus response. In contrast, nitrogen concentration had little effect on photochemical efficiency. Changes in biovolume over time were used to compare growth rates of four accessions in response tonitrogen and phosphorus supply. We demonstrate that a time series image-based approach coupled with mathematical modeling provides higher resolution of genotypic response to nutrient supply than traditional destructive techniques and shows promise for high throughput screening and determina- tion of genomic regions associated with superior nutrient use efficiency.     

The key residue for SSB-RecO interaction is dispensable for Deinococcus radiodurans DNA repair in vivo

The RecFOR DNA repair pathway is one of the major RecA-dependent recombinatorial repair pathways in bacteria and plays an important role in double-strand breaks repair. RecO, one of the major recombination mediator proteins in the RecFOR pathway, has been shown to assist RecA loading onto single-stranded binding protein （SSB） coated single-stranded DNA （ssDNA）. However, it has not been characterized whether the protein-protein interaction between RecO and SSB contributes to that process in vivo. Here, we identified the residue arginine-121 of Deinococcus radiodurans RecO （drRecO-R121） as the key residue for RecO-SSB interaction. The substitution of drRecO-R121 with alanine greatly abolished the binding of RecO to SSB but not the binding to RecR. Meanwhile, SSB-coated ssDNA annealing activity was also compromised by the mutation of the residue of drRecO. However, the drRecO-R121A strain showed only modest sensitivity to DNA damaging agents. Taking these data together, arginine-121 of drRecO is the key residue for SSB-RecO interaction, which may not play a vital role in the SSB displacement and RecA loading process of RecFOR DNA repair pathway in vivo.     

High-accuracy mapping of protein binding stability on nucleosornal DNA using a single-molecule method

Dear Editor, The conformation of nucleosomal DNA is significantly different from that of B-form double stranded DNA （dsDNA） （Richmond and Davey, 2003）. In nucieosomal DNA, specific DNA sequences are less flexible and less accessible than in free dsDNA, which might be due to the tight association of histone cores. The allosteric effect via DNA has been documented recently （Kim et al., 2013）, suggesting that DNA is not merely a solid rod providing recognition sequences. Previous studies of nucleosomal DNA-protein interactions only demonstrated the key mechanism involved--histone shielding （Li and Wrange, 1993; Hinz et al., 2010; Sahu et al., 2010）.     

pIRES2-GDNF-NT-3真核表达载体的构建与鉴定(英文)

目的:采用一种简便和高效的方法构建双基因共表达载体pIRES2-GDNF-NT-3。方法:人胶质细胞源性神经营养因子和神经营养素3是采用PCR的方法从人外周血单个核细胞的基因组DNA中获取,将人胶质细胞源性神经营养因子的cDNA片段插入到pIRES2-EGFP多克隆位点构建成为pIRES2-GDNF-EGFP.神经营养素3 cDNA片段通过替换EGFP的方式插入到pIRES2-GDNF-EGFP中构建成为pIRES2-GDNF-NT-3双基因共表达载体。结果:人胶质细胞源性神经营养因子和神经营养素3被克隆,通过测序和酶切鉴定的得知与基因库报道序列一致。结论:人神经生长因子和神经营养素3双基因真核表达载体成功构建,它提供了一个新的表达系统,为进一步研究双基因的功能奠定了基础。     

原发性肝癌患者术后两种营养支持的疗效对比及对免疫功能的影响研究

目的:研究原发性肝癌患者术后两种营养支持的疗效及对免疫功能的影响。方法:病例选自2010年10月至2012年11月在我院就诊治疗的90例经诊断为原发性肝癌并进行肝部分切除的患者,随机分为EN组和PN组,每组各45例,分别进行肠内和肠外营养支持治疗,观察术前及术后患者的营养状况、住院时间、肠道功能恢复时间、并发症情况及免疫功能的变化,经统计学处理,探讨两种不同营养支持疗法的临床疗效及对免疫功能的影响。结果:相对于PN组,EN组患者术后营养明显改善,胃肠道功能恢复相对较快,免疫功能明显提高,术后并发症明显减少,且P0.05。差异具有统计学意义。结论:早期EN比PN更能改善肝癌病人术后肠道功能及营养状况,降低术后并发症,提高患者的免疫功能,有利于患者术后恢复。