Selection of reference genes for quantitative real-time PCR normalisation in largemouth bass Micropterus salmoides fed on alternative diets

The partial cDNA sequences of eight reference genes (actb, tuba1, gapdh58, gapdh59, eef1a1, RNA 18 s, pabpc1, ube2I) were cloned from largemouth bass Micropterus salmoides. The expression levels of these eight genes were compared in the various tissues (eye, spleen, kidney, gill, muscle, brain, liver, heart, gut and gonad) of M. salmoides fed on forage fish. The results showed that the candidate genes exhibited tissue-specific expression to various degrees and the stability ranking order was eef1a1 > tuba1 > RNA 18 s > pabpc1 > ube2I > actb > gapdh58 > gapdh59 among tissue types. Four candidate genes eef1a1, tuba1, RNA 18 s and actb were used to analyse the stability in liver tissues of largemouth bass between the forage-fish group and the formulated-feed group. The candidate genes also showed some changes in expression levels in the livers, while eef1a1 and tuba1 had the most stable expression in livers of fish fed on alternative diets within 10 candidates. So eef1a1 and tuba1 were recommended as optimal reference gene in quantitative real-time PCR analysis to normalise the expression levels of target genes in tissues and lives of the M. salmoides fed on alternative diets. In livers, the expression levels of gck normalised by eef1a1 and tuba1 showed the significant up-regulation in formulated feed group (P < 0.05) than those in forage-fish group. While sex difference has no significant effects on the expression levels of gck in both groups.     

A 55 K SNP array-based genetic map and its utilization in QTL mapping for productive tiller number in common wheat

Key message

A high-density genetic map constructed with a wheat 55 K SNP array was highly consistent with the physical map of this species and it facilitated the identification of a novel major QTL for productive tiller number.

Abstract

Productive tiller number (PTN) plays a key role in wheat grain yield. In this study, a recombinant inbred line population with 199 lines derived from a cross between ‘20828’ and ‘Chuannong16’ was used to construct a high-density genetic map using wheat 55 K single nucleotide polymorphism (SNP) array. The constructed genetic map contains 12,109 SNP markers spanning 3021.04 cM across the 21 wheat chromosomes. The orders of the genetic and physical positions of these markers are generally in agreement, and they also match well with those based on the 660 K SNP array from which the one used in this study was derived. The ratios of SNPs located in each of the wheat deletion bins were similar among the wheat 9 K, 55 K, 90 K, 660 K and 820 K SNP arrays. Based on the constructed maps, a novel major quantitative trait locus QPtn.sau-4B for PTN was detected across multi-environments in a 0.55 cM interval on 4B and it explained 17.23–45.46% of the phenotypic variance. Twenty common genes in the physical interval between the flanking markers were identified on chromosome 4B of ‘Chinese Spring’ and wild emmer. These results indicate that wheat 55 K SNP array could be an ideal tool in primary mapping of target genes and the identification of QPtn.sau-4B laid a foundation for the following fine mapping and cloning work.

     

Stoichiometry of leaf nitrogen and phosphorus of grasslands of the Inner Mongolian and Qinghai-Tibet Plateaus in relation to climatic variables and vegetation organization levels

Nitrogen (N) and phosphorus (P) are most commonly the limiting essential elements that affect the functioning of plants and ecosystems. However, their stoichiometry in relation to climatic variables and vegetation organization levels has not been comprehensively characterized. N and P concentrations were measured for 329 leaf samples collected at 132 sites along the 5000 km long China Grassland Transect that traverses the Inner Mongolian and Qinghai-Tibet Plateaus. The patterns of these measurements were analyzed with reference to climate factors, plant species, plant functional groups, grassland communities and grassland ecosystems. The aim was to explore whether geographical patterns of plant leaf elements are related to zonal climatic variables, and at which vegetation organization levels changes of plant leaf N and P stoichiometric characteristics and pattern occur. Results showed that interspecific differences of N and P concentrations were most significant for the three vegetation organization levels of species, community and ecosystem. Plant leaf N and P concentrations were higher, coefficients of variation of N and P lower, and N/P, C/N and C/P ratios were also lower for leaf samples from the cold high altitude Qinghai-Tibet Plateau than for those from the relatively lower altitude and warmer Inner Mongolian Plateau. Correlation of N and P for Inner Mongolian grassland was higher than that for the Qinghai-Tibet Plateau. The study indicates plant species are the most basic unit influencing plant stoichiometric geographic patterns, and that climatic variables affect leaf element concentrations mainly through their effect on changes of plant species composition of vegetation.     

利用Tol2转座子构建斑马鱼心脏组织特异表达转基因载体及其表达分析

为了制备用于在斑马鱼心脏中特异表达目的基因的转基因载体,通过分子克隆的方法对能够在斑马鱼心脏中特异表达EGFP报告基因的Tol2载体进行了改造,在原有的CMLC2启动子与EGFP编码区之间插入带有多克隆位点的IRES序列,获得pTol2-CMLC2-IRES-EGFP转基因表达载体,该载体可以实现在同一个启动子CMLC2的驱动下分别同时表达目的基因和EGFP;为了验证该表达载体的有效性,进一步在CMLC2启动子与IRES序列之间插入DsRed-Monome编码区,利用得到的pTol2-CMLC2-RED-IRES-EGFP转基因载体显微注射到斑马鱼单细胞期胚胎中进行表达分析,结果表明外源目的基因DsRed-Monome和报告基因EGFP均能以相同的表达模式在斑马鱼心脏组织中特异表达。pTol2-CMLC2-IRES-EGFP转基因表达载体的成功构建对于建立心脏发育候选基因的斑马鱼转基因实验模型具有重要意义。     

ABA调控种子发育的研究进展

种子发育是一个复杂的生物学过程，受各种遗传和外界因素的调节，显著影响农作物特别是禾谷类作物的种子活力和产量与质量。脱落酸(ABA)是调控种子发育和萌发最重要的植物激素之一，其活性水平、信号转导及其LAFL网络在种子发育包括胚胎发生和成熟过程的调控中起关键作用。该文主要综述了近年来ABA调控种子发育的研究取得的重要进展，包括ABA代谢和信号转导对种子发育的调控，ABA与种子成熟转录因子(AFL-B3、FUS3、ABI3、LEC2等)的作用，以及ABA在种子发育中的作用机制，并提出了需要进一步研究的科学问题，为深入理解种子发育的分子机制提供参考，从而提高种子的活力、产量和质量。     

川东红池坝地区红三叶和鸭茅人工草地土壤和植物营养元素含量特征的研究

川东地区池坝地区红三叶（Ｔｒｉｆｏｌｉｕｍｐｒａｔｅｎｓｅ）和鸭茅（Ｄａｃｔｙｌｉｓｇｌｏｍｅｒａｔａ）人工草地土壤和植物营养元素的含量特征如下;（１）土壤中的元素含量以铁,钾和镁较高,钠,钙,氮,锰和磷较低,硫,锌,硼,铜和钼微少;（２）从元素的富集特征来看,该区土壤中的钙,硫为重度淋溶元素,钾,磷,镁,锌,钠为中度淋溶元素,铁,铜属轻度淋溶元素,锰属富集元素;（３）根据元素的生物吸收系列,红三     

乳腺癌组织中ER、PR和CEA相关性研究

应用免疫组化技术亲和组化法和ＡＢＣ法,检测了１０６例乳腺癌组织中ＥＲ、ＰＲ和ＣＥＡ水平。其阳性率依次为８３％、８１．１％和８８．７％。其中７９．２％的乳腺癌ＥＲ和ＰＲ表达一致。在９８例ＥＲ和／或ＰＲ阳性乳腺癌中有９２例（９３．９％）呈ＣＥＡ阳性,８例ＥＲ和ＰＲ阴性乳腺癌中６例（７５％）为ＣＥＡ阴性,８９％乳腺癌ＣＥＡ与ＥＲ和ＰＲ表达一致。在癌的分级表达中,随着癌的组织学分级增高ＣＥＡ阳性率增高,而ＥＲ和ＰＲ阳性率减低。结果表明,乳腺癌ＥＲ、ＰＲ和ＣｈＡ表达可反映肿瘤的不同生物学特征。同时检测三者对选择内分泌治疗及判断预后更有意义。     

Detection and control of aspartimide formation in the synthesis of cyclic peptides

Extensive two-dimensional NMR analysis was employed to characterize the structural identity of the macrocyclic peptide lactam and the imide analog, a major side reaction product when allyl ester was used to protect the side chain of aspartic acid. A straightforward protocol modification was developed to minimize aspartimide formation during the synthesis of cyclic peptides.     

Efficacy of Oral Cryotherapy on Oral Mucositis Prevention in Patients with Hematological Malignancies Undergoing Hematopoietic Stem Cell Transplantation: A Meta-Analysis of Randomized Controlled Trials

Objectives

Controversy exists regarding whether oral cryotherapy can prevent oral mucositis (OM) in patients with hematological malignancies undergoing hematopoietic stem cell transplantation (HSCT). The aim of the present meta-analysis was to evaluate the efficacy of oral cryotherapy for OM prevention in patients with hematological malignancies undergoing HSCT.

Methods

PubMed and the Cochrane Library were searched through October 2014. Randomized controlled trials (RCTs) comparing the effect of oral cryotherapy with no treatment or with other interventions for OM in patients undergoing HSCT were included. The primary outcomes were the incidence, severity, and duration of OM. The secondary outcomes included length of analgesic use, total parenteral nutrition (TPN) use, and length of hospital stay.

Results

Seven RCTs involving eight articles analyzing 458 patients were included. Oral cryotherapy significantly decreased the incidence of severe OM (RR = 0.52, 95% CI = 0.27 to 0.99) and OM severity (SMD = -2.07, 95% CI = -3.90 to -0.25). In addition, the duration of TPN use and the length of hospitalization were markedly reduced (SMD = -0.56, 95% CI = -0.92 to -0.19; SMD = -0.44, 95% CI = -0.76 to -0.13; respectively). However, the pooled results were uncertain for the duration of OM and analgesic use (SMD = -0.13, 95% CI = -0.41 to 0.15; SMD = -1.15, 95% CI = -2.57 to 0.27; respectively).

Conclusions

Oral cryotherapy is a readily applicable and cost-effective prophylaxis for OM in patients undergoing HSCT.