5种蔷薇科树种多酚氧化酶比较基因组学分析

蔷薇科植物中有多种具有重要价值的水果,如苹果、梨、桃、草莓和黑树莓等。这些水果普遍容易发生由多酚氧化酶(PPO)介导的酶促褐变而导致重大经济损失,从全基因组角度分析比较PPO基因家族,有助于加深对PPO基因家族和功能的认识。采用比较基因组学的方法,对这5种植物的PPO基因家族进行了基因鉴定、染色体定位、编码蛋白的亚细胞定位、内含子统计分析、基因系统进化、基因倍增与丢失等特征分析。从这5种植物中,共计鉴定出了42个PPO基因,其中6个基因被认定是假基因;绝大多数基因编码的蛋白定位于叶绿体,2个定位于线粒体,3个是分泌型蛋白;PPO基因在染色体上有串联重复和散布两种形式;9个基因具有内含子,聚类结果显示可以将它们分为6个类型,每个基因类型在进化过程中都发生过基因倍增和丢失;同型基因内含子的位置和大小具有相似性。这些结果揭示蔷薇科植物PPO基因内含子是伴随着基因倍增产生的,基因倍增也是推动PPO基因多样化的重要动力,PPO基因倍增和丢失差异导致PPO基因数量在不同物种之间产生差异。     

A Genome-Wide Association Study Reveals Differences in the Genetic Mechanism of Control of the Two Gait Patterns of the Brazilian Mangalarga Marchador Breed

The DMRT3 gene is described as the main gene involved in the determination of gait phenotypes in horses, and the allele A of the 22999655C>A single nucleotide polymorphisms (SNP) has been reported as a causal variant of this trait. In the Mangalarga Marchador breed, which exhibits two gait patterns with well-defined characteristics, genotypes AA and CA are associated with marcha picada and genotype CC with marcha batida. In this breed, allele A of the DMRT3 gene is only related to the marcha picada gait. The objective of this study was to identify the type of control of the marcha batida gait and to investigate SNPs and genomic regions responsible for this phenotype in Mangalarga Marchador horses. Forty-eight horses belonging to the two gait groups, marcha picada with AA and CA genotypes of the 22999655C>A SNP (n = 20) and marcha batida with CC genotype (n = 28), were analyzed using the Equine SNP70 BeadChip. The genome-wide association study result shows for the first time that, in contrast to the marcha picada gait phenotype that is apparently determined by a single gene (DMRT3) in which allele A of variant g.22999655C>A controls the trait, the marcha batida gait is controlled by a larger number of genes. Because of the small number of animals used in the two groups compared, the genomic regions associated with smaller effects on the marcha batida gait could not be identified.     

葡萄WRKY54基因克隆及表达特性分析

为了探究葡萄WRKY54基因的功能,以抗盐葡萄品种Vidal Blanc为材料,采用同源克隆法克隆得到Vv WRKY54基因,并对其进行生物信息学和表达特性分析。结果表明,Vv WRKY54基因c DNA序列为942 bp,编码313个氨基酸。生物信息学分析结果表明,Vv WRKY54蛋白分子量约为35.3091 k Da,等电点为5.45,不稳定系数为55.03,推测其为不稳定蛋白,与已知毛果杨及拟南芥WRKY54蛋白高度同源;亚细胞定位预测结果显示其主要存在于细胞核中。实时荧光定量PCR分析表明,Vv WRKY54在葡萄不同组织中均有表达,其中在梢尖中表达量最高;盐和低温等逆境胁迫因子均能诱导Vv WRKY54上调表达;此外,Vv WRKY54受水杨酸和一氧化氮诱导上调表达,其中水杨酸诱导Vv WRKY54相对表达量在12 h达到最大值,约为对照的50倍。推测Vv WRKY54在植物发育和抵御逆境胁迫中起着重要作用,这为进一步阐明Vv WRKY54的功能及作用机制奠定了分子基础。     

香蕉枯萎病菌4号生理小种cat1基因敲除与表型分析

前期蛋白质组学研究发现尖孢镰刀菌古巴专化型4号生理小种(Foc4)中的过氧化氢酶1(cat1)基因受到巴西蕉诱导上调表达,为研究cat1基因在Foc4侵染香蕉过程中的作用,利用同源重组方法敲除Foc4的cat1基因,并对获得的敲除突变体开展表型分析和致病性测定。结果表明:cat1基因缺失对Foc4的菌丝、孢子形态、菌落生长、抗渗透压胁迫和细胞壁选择性压力等没有影响;但引起菌株抵抗外源氧胁迫、细胞壁穿透能力、纤维素利用能力减弱和对巴西蕉的致病性减弱。显示cat1基因的产物通过清除香蕉分泌的活性氧在Foc4对香蕉的致病过程中起作用,并参与病原菌对寄主纤维素成分的代谢。此结果为进一步深入研究香蕉枯萎病菌的致病机理奠定基础。     

Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus)

Animal digestive tract is habitat for a large number of autochthonous microbiota, which play central roles in multiple biological and physiological processes of the host. In this study, two different micro‐biomass preparation methods were employed to evaluate the diversity of intestinal mucosa‐associated microbiota in grass carp (Ctenopharyngodon idellus). Genomic DNAs were isolated either directly from intestinal mucosal samples (group A), or from micro‐biomass after microbial dissociation (group B). Community richness, diversity and evenness indices were all higher in group B, but differences were not statistically significant (P = 0.97, P = 0.33, P = 0.34 respectively). Furthermore, group B samples exhibited an increased ratio of bacterial DNA in comparison with group A samples, but the difference was also not statistically significant (P = 0.74). In addition, there were no statistically significant differences between the two groups (P > 0.05) at the taxonomic level. Our results support previous findings that there exists a great abundance of the intestinal mucosa‐adherent microbiota in the grass carp; among these, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetes and Fusobacteria were the most common phyla. Within these microbiota, Paenibacillus, Bacteroides, Bacillus and Cetobacterium genera comprise the majority of the community, implicating their functional importance (e.g. as probiotics) to their host. Our results contribute towards a better understanding of the intestinal microbial profile of grass carp. Both micro‐biomass preparation techniques proved to be feasible for studying mucosa‐adherent microbiota of grass carp; however, the second method (group B) provides a protocol that is somewhat more effective than the first method (group A).     

福瑞鲤选育家系不同养殖阶段的生长差异分析

为了进一步观察最佳线性无偏预测(best linear unbiased prediction,BLUP)家系选育方法在福瑞鲤(Cyprinus carpio)继代选育中的潜力,该研究测量了继续选育第2代家系群体不同养殖阶段的体质量和形态性状。结果表明,生长快速家系群福瑞鲤早期(4月龄)生长速度较慢,到后期则生长加快,其体质量增长表现出明显的优势。在体型方面,随着养殖时间的延长,福瑞鲤各选育家系群的体厚/体长增加,体高/体长降低,逐渐呈现其体型修长的特征;同时2个越冬期的成活率均达到了94%以上。结果表明通过BLUP家系选育对福瑞鲤长期选育是可行的。在此基础上,通过主成分分析发现,福瑞鲤生长性状第一主成分是体质量;对不同生长时期的体质量进行相关性分析,发现9月龄、14月龄、21月龄鱼的体质量与24月龄的相关系数均达到极显著水平(P0.01),分别为0.851、0.897和0.957。因此,在福瑞鲤继续选育过程中,进行早期个体选择值得尝试。     

Can water disinfection prevent the transmission of infectious koi herpesvirus to naïve carp? – a case report

Hygienic measures such as disinfection are important tools for the maintenance of fish health in aquaculture. While little information is available on the disinfection of water intended for fish containment, Huwa‐San^®, a disinfectant used in food and water industries, was used for daily treatment at concentrations of approximately 60 ppm over a total period of 3 months (experiment 1) with a 3‐week treatment‐free interval after 2 months (experiment 2). During this period, koi herpesvirus (KHV) was added to the water of two aquaria, one used as a normal contact control, the other one receiving daily water disinfectant treatments that prevented KHV infection of carp. In the second experiment, Huwa‐San^® treatment was interrupted and KHV infection was prevalent. However, when naïve fish were introduced to the same aquarium after re‐application of disinfectant, KHV could not be detected in those naïve fish. Whilst KHV could not be detected in samples where disinfectant had been applied, it was present in samples of naïve fish cohabiting with infection contact control animals which had undergone no disinfectant treatment over experiments 1 and 2. The results presented here show that water treatment with a disinfectant may prevent transmission of infectious KHV to naïve carp cohabited with infected carp.     

茶树几丁质酶基因的克隆及其在干旱胁迫下的表达分析在干旱胁迫下的表达分析

以铁观音茶树叶片为材料,利用逆转录PCR及RACE法,克隆了茶树几丁质酶基因CsChi(GenBank登录号为KR078345).CsChi基因的cDNA全长为1 192 bp,包含972 bp的开放阅读框(ORF),编码323个氨基酸.生物信息学分析结果表明,CsChi蛋白的分子量为34.33 ku;理论等电点pI为8.44;原子组成为C1519H2285N413O464S18,总原子数为4 699;蛋白质结构分析显示该蛋白有6个蛋白的跨膜区域,属于跨膜蛋白;存在于细胞外;没有卷曲螺旋结构存在;CsChi基因编码的蛋白属于糖苷水解酶19家族,含有保守的ChtBD1结构域,与溶菌酶的保守结构域类似,可能兼具几丁质酶活性和溶菌酶活性,qPCR定量分析结果显示在不同干旱胁迫处理下茶树的CsChi基因的表达量,与对照组相比有所增加.推测CsChi基因在茶树干旱等逆境胁迫中起重要作用.     

Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

三氯生对红白鲫运动行为的影响

利用视频跟踪系统研究三氯生(TCS)对红白鲫运动行为的影响。将红白鲫(red-white crucian carp)暴露在5种不同浓度的TCS(0.50、0.60、0.70、0.80和0.90 mg/L)水体中,视频记录5 min内红白鲫的运动轨迹(运动轨迹线、摆动次数)、游动速度和垂直穿梭次数的变化。同时,进行TCS(0.60、0.69 mg/L)对红白鲫的14 d暴露实验,暴露实验结束后,2 min内监测上述指标。结果显示:短期TCS暴露,红白鲫的运动轨迹条数、摆动次数、垂直穿梭次数、运动速率显著增加;暴露14 d后的红白鲫上述运动行为学指标显著降低。结果表明:在一定浓度范围内,TCS能够改变红白鲫的运动行为。