茶树CsLHTs亚家族基因的克隆与表达分析

氨基酸是茶叶的重要品质成分和氮素贮存形式,因此开展茶树体内氨基酸转运蛋白的研究尤为重要。本研究从茶树转录组中筛选得到5条茶树LHTs(Lysine histidine transporters,赖氨酸/组氨酸转运蛋白)家族基因序列,并从龙井43中成功克隆获得4个茶树LHTs基因,分别命名为CsLHT1、CsLHT6、CsLHT8.1和CsLHT8.2。对该亚家族编码的氨基酸序列的理化性质和功能结构进行预测,结果显示CsLHTs亚家族含有9~11个跨膜区;且含有与氨基酸转运相关的结构域。为了明确在不同茶树品种和氮素水平间CsLHTs基因的表达差异,本研究选用3个茶树品种的扦插苗为试验材料,水培条件下氮饥饿两周后,分别用0.2、2、10mmol·L^-1的NH4NO3进行处理。利用qRT-PCR分析了CsLHTs在不同组织间的表达情况,结果表明4个基因在茶树营养组织中均有表达。经氮素处理后,CsLHTs基因在3个氮素水平和3个品种中呈现出不同的变化规律。CsLHT1、CsLHT6和CsLHT8.2在品种间的表达差异程度高于不同氮素水平间的基因表达差异。CsLHT8.1对氮素处理有明显的响应,尤其经0.2mmol·L^-1和10mmol·L^-1的NH4NO3处理72h后,在氮高效品种中茶302根中呈上调表达,表明CsLHT8.1可能参与氨基酸由根部向地上部的转运过程。     

Traits-related QTLs and genes and their potential applications in rice improvement under low phosphorus condition

The genetic improvement of rice over past decades has led to the loss of several genes which are responsible for nutrient acquisition and soil-related stresses. Phosphorus (P) is a non-renewable resource and inevitable element of metabolic functions in plants. By pedogenesis process, organic matter contributes to renewing soil P in lesser extent. Therefore, improvement of the P use efficiency is one of the most imperative traits in rice breeding program, which is governed by quantitative trait loci (QTLs). QTLs controlling low P associated morphological and physiological traits in rice were investigated mostly during last one and half decade by using diverse mapping populations. Comprehensive and meticulous survey from literature, we found that to date 133 P associated QTLs of morpho-physiological traits were reported to be distributed on twelve chromosomes and majority of these QTLs localized on chromosome 1, 2 and 12. For the first time, a complete figure is presented in this review on chromosome wise with respective QTLs associated with low P for easy understanding and selecting markers for future prospect. Further, it is necessary to understand the molecular mechanisms and regulation of genes and traits associated with low P to develop tolerant rice cultivars using functional marker-assisted selection.     

Effects of cysteamine supplementation on the intestinal expression of amino acid and peptide transporters and intestinal health in finishing pigs

This study aimed to evaluate the effects of cysteamine supplementation on the expression of jejunal amino acid and peptide transporters and intestinal health in finishing pigs. Sixty barrows were allocated into two experimental diets consisting of a basal control diet supplemented with 0 or 142 mg/kg cysteamine. After 41 days, 10 pigs per treatment were slaughtered. The results showed that cysteamine supplementation increased the apparent digestibility of crude protein (CP) (P < 0.05) and the trypsin activity in jejunal digesta (P < 0.01). Cysteamine supplementation also increased the messenger RNA abundance of SLC7A7, SLC7A9 and SLC15A1, occludin, claudin‐1 and zonula occludens protein‐1 (P < 0.001) in the jejunum mucosa. Increased glutathione content (P < 0.01) and glutathione peroxidase activity (P < 0.05) and decreased malondialdehyde content (P < 0.01) were observed in pigs receiving cysteamine. Additionally, cysteamine supplementation increased the concentrations of secretory immunoglobulin A (IgA) (P < 0.05), IgM (P < 0.001) and IgG (P < 0.001) in the jejunal mucosa. It is concluded that cysteamine supplementation could influence protein digestion and absorption via increasing trypsin activity, enhancing the digestibility of CP, and promoting the expression of jejunal amino acid and peptide transporters. Moreover, cysteamine improved intestinal integrity, antioxidant capacity and immune function in the jejunum, which were beneficial for intestinal health.     

Canine ABCB4: Tissue expression and cDNA structure

The ABCB gene subfamily of ABC (ATP-binding cassette) transporters is responsible for transporting a wide spectrum of molecules including peptides, iron, bile salts, drugs, and phospholipids. In humans, ABCB4 appears to be exclusively expressed on the apical membrane of hepatocytes where it translocates phosphatidylcholine from the inner to the outer leaflet of the canalicular membrane. Functional alterations in the ABCB4 transporter are associated with a number of cholestatic syndromes in humans. Because of its role in biliary lipid homeostasis in humans, investigation of the ABCB4 gene in dogs is warranted. Thus, the full cDNA sequence of canine ABCB4 was elucidated and its mRNA and protein expression levels in tissues were determined. Canine ABCB4 consists of 3804 nucleotides spanning 26 exons and is 89% identical to human ABCB4. Expression of ABCB4 in canine liver supports a potential role for the protein in normal biliary function similar to that in humans. The function of ABCB4 expressed in brain tissue has yet to be determined.     

Degree of Suppression of Mouse Myoblast Cell Line C2C12 Differentiation Varies According to Chondroitin Sulfate Subtype

Chondroitin sulfate (CS), a type of glycosaminoglycan (GAG), is a factor involved in the suppression of myogenic differentiation. CS comprises two repeating sugars and has different subtypes depending on the position and number of bonded sulfate groups. However, the effect of each subtype on myogenic differentiation remains unclear. In this study, we spiked cultures of C₂C₁₂ myoblasts, cells which are capable of undergoing skeletal muscle differentiation, with one of five types of CS (CS-A, -B, -C, -D, or -E) and induced differentiation over a fixed time. After immunostaining of the formed myotubes with an anti-MHC antibody, we counted the number of nuclei in the myotubes and then calculated the fusion index (FI) as a measure of myotube differentiation. The FI values of all the CS-treated groups were lower than the FI value of the control group, especially the group treated with CS-E, which displayed notable suppression of myotube formation. To confirm that the sugar chain in CS-E is important in the suppression of differentiation, chondroitinase ABC (ChABC), which catabolizes CS, was added to the media. The addition of ChABC led to the degradation of CS-E, and neutralized the suppression of myotube formation by CS-E. Collectively, it can be concluded that the degree of suppression of differentiation depends on the subtype of CS and that CS-E strongly suppresses myogenic differentiation. We conclude that the CS sugar chain has inhibitory action against myoblast cell fusion.     

苏云金芽胞杆菌类ABC transporter基因zwa-FEG能提高双效菌素的抗性

双效菌素(zwittermicin A,ZwA)是一种氨基多元醇类新型广谱抗生素,zmaR为其抗性基因。在苏云金芽胞杆菌(Bacillus thuringiensis)YBT-1520菌株ZwA合成基因簇末端存在一个类似ABC transporter的序列(命名为zwa-FEG)。将zwa-FEG基因在大肠杆菌(Escherichia coli)中表达,大肠杆菌能获得ZwA的抗性;将zwa-FEG转移到产ZwA的蜡状芽胞杆菌(Bacillus cereus)菌株UW85中,能使其ZwA产量显著提高。推测zwa-FEG是一种ZwA专一性的ABC transporter,它能将ZwA分泌到胞外,从而增强ZwA的合成和ZwA的抗性。     

鸡脑催乳素免疫反应神经元的分布

运用ＡＢＣ法观察了催乳素（ＰＲＬ）免疫反应神经元在鸡脑的分布。结果,ＰＲＬ阳性胞体主要分布在视交叉上核、视前室旁核、视上核、下丘脑室旁核、弓状核和伏隔核,旧纹状体、正中隆起存在大量阳性纤维末梢,在侧脑室腹侧的室管膜和脑基底神经胶质板上也存在ＰＲＬ阳性神经元。视前室旁核、视交叉上核、下丘脑室旁核、弓状核等核团内的ＰＲＬ阳性神经元有突起向第三脑室投射,伏隔核内及侧脑室室管膜上的ＰＲＬ阳性神经元有突起伸至侧脑室,视上核的ＰＲＬ阳性神经元也有突起投射至脑基底神经胶质板,表明鸡脑内的ＰＲＬ可以释放入脑室系统,参与调节脑－脑脊液神经体液回路。     

The differences in postprandial free amino acid concentrations and the gene expression of PepT1 and amino acid transporters after fishmeal partial replacement by meat and bone meal in juvenile turbot (Scophthalmus maximus L.)

Meat and bone meal (MBM) is a high‐quality alternative protein source used to replace fishmeal (FM). However, the molecular mechanisms of over‐substituted FM by MBM resulted in growth reduction are still not clear. The objective of the study was to evaluate the effect of FM replacement by MBM on the concentration of postprandial free amino acid (FAA) and mRNA abundance of peptide and amino acid transporters in juvenile turbot (Scophthalmus maximus L.). Fish were fed with FM diet (60% FM), MBM diet (33% FM + 34.2% MBM) and MBM + AA diet (MBM diet with essential amino acid (EAA) added to match the AA profile of FM) for 30 days. Results showed that compared with the FM diet, MBM diet led to a reduction in FAA concentration peak values in plasma and muscle. MBM + AA diet significantly elevated the peak values of FAA concentrations to FM diet level in plasma, but not in muscle. Furthermore, compared with FM diet, MBM diet significantly increased gene expression of PepT1 and major amino acid transporters in intestine, whereas MBM diet greatly downregulated gene expression of T‐type amino acid transporter‐1, system ASC amino acid transporter‐2 and cationic amino acid transporter‐2 in muscle. Supplemented EAA did not ameliorate these different effects in intestine and muscle. Overall, this study provided a comprehensive explanation for the relationship between diet, FAA concentrations and AA transportations, which provides a molecular basis for further using MBM to replace FM in aquafeeds.     

七星列岛海域春秋季游泳动物群落特征

基于2014年秋季和2015年春季七星列岛海域的底拖网渔获物进行分析,研究七星列岛海域游泳动物的种类组成、优势种、多样性、生态位和群落结构特征,得出游泳动物共80种,隶属于3纲14目42科62属,其中鱼类52种,虾类13种,蟹类10种,头足类5种。秋季优势种有6种,春季有4种,共有种为三疣梭子蟹。两季物种多样性指数(H')、(D)、(J')均为中等水平。生态位数值较大的物种秋季有六指马鲅(10.51)、哈氏仿对虾(9.47)、海鳗(8.84)等,春季有火枪乌贼(8.93)、口虾蛄(8.51)、带鱼(7.94)等。聚类分析显示,两季物种站位均可分为3组,与NMDS分析结果一致。ABC曲线分析得出,W值秋季为0.052、春季为–0.059,均接近0。该海域游泳动物群落组成以鱼类为主,虾类次之,头足类最少,优势种变化明显;哈氏仿对虾、海鳗、口虾蛄等低价值游泳动物资源生态位数值高,占据主要竞争地位;由于组内站位间物种组成和海域环境有一定程度的相似性,两季物种站位间分组不受季节影响。ABC曲线分析显示该海域处于中度干扰状态。