斑节对虾血淋巴低丰度蛋白双向电泳技术体系的优化

文章探索并优化了斑节对虾（Penaeus monodon）血淋巴双向电泳技术体系,对斑节对虾血淋巴总蛋白抽提方法进行优化,减小高丰度蛋白的影响;对双向电泳过程各步骤,如IPG胶条pH范围的选择、等电聚焦程序、上样量等进行了优化,得到高分辨率的2-DE图谱,用PDQuest软件进行了初步分析。结果表明,用PEG 6000能更快捷有效地去除血淋巴中的高丰度蛋白,增加低丰度蛋白的点数及浓度;在抽提蛋白过程中,利用预冷的80%丙酮洗涤,在一向电泳过程中设置100 V低压除盐,能更有效地去除蛋白中的盐分;采用18 cm、pH 3~10 NL 的中型胶条,能更好地显示低丰度蛋白点的分布;被动水化上样300 μg结合硝酸银染色方法,能有效提高双向电泳图谱中蛋白点的分离度和分辨率。该试验为斑节对虾及其他甲壳动物血淋巴蛋白质组学相关研究提供了稳定可重复的研究方法。     

不同采收期咖啡果实物理性状比较

以潞江坝的Catimor系列品种咖啡鲜果为材料,研究不同采收期对咖啡果实物理性状的影响。结果表明,不同成熟期果实的物理性状差异为极显著,其中,早期和晚期成熟的咖啡鲜果和咖啡豆颗粒较小,物理性状较差,而中期成熟的咖啡鲜果和咖啡豆颗粒较大,物理性状较好。     

Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus (Bloch 1790)

Ontogenetic development of digestive system in crimson snapper (Lutjanus erythopterus Bloch 1790) larvae was histologically and enzymatically (alkaline phosphatase, amylase, lipase, pepsin, trypsin) examined from hatching to 36 days post hatching (DPH). The ontogenetic development of crimson snapper larval fish ontogeny was divided into three distinct phases: phase I starting from hatch to the onset of exogenous feeding, phase II starting from first feeding (2–3 DPH) until the formation of gastric glands (13–14 DPH) and phase III beginning from the appearance of gastric glands and continuing onwards. The specific activities of amylase, lipase, trypsin showed sharp increase and reached to the maximum from hatch to 4 DPH, 10 DPH, 20 DPH, respectively, followed by a declining trend with irregular fluctuation. In contrast to other enzymes, the specific activities of alkaline phosphatase showed a gradual increase from hatch to 29 DPH, followed by a sharp increase towards 36 DPH. The specific activity of pepsin was firstly detected on 17 DPH and gradually increased towards the end of this study. The total activities of these five enzymes showed a gradual increase till 29 DPH, followed by a sharp increase towards 36 DPH except for amylase and lipase reaching maximum at 32 DPH. The present study provides better understanding of the digestive ontogeny of crimson snapper during the larval stage and a guide to feeding and weaning of this economically important fish in hatcheries.     

斑节对虾含硒谷胱甘肽过氧化物酶基因全长克隆及表达分析

为了研究含硒谷胱甘肽过氧化物酶(selenium-dependent glutathione peroxidase,SeGPx)在斑节对虾应激反应和卵巢发育中的作用,实验以斑节对虾肝胰腺转录组数据中筛选获得的Se-GPx基因(Pm Se-GPx)片段为基础,利用RACE技术获得Pm Se-GPx基因的c DNA全长,并对其进行了生物信息学分析;利用荧光定量PCR技术研究了Pm Se-GPx在斑节对虾不同组织、卵巢发育各期、肝胰腺组织在pH9、硫酸铜(Cu~(2+))应激下的相对表达量和变化趋势。结果显示,Pm Se-GPx基因c DNA全长为959 bp,5'非编码区(UTR)长10bp,开放阅读框(ORF)长639 bp,编码212个氨基酸,310 bp的3'UTR包含一个硒代半胱氨酸插入序列(SECIS),ORF中的密码子209TGA211编码硒代半胱氨酸(selenocysteine,U67)。实时定量PCR实验结果表明,Pm Se-GPx在斑节对虾的肝胰腺、卵巢、精巢、心脏等组织中均有表达,其中肝胰腺中表达量显著高于其他组织,卵巢次之;卵巢发育阶段表达结果则显示Pm Se-GPx在卵巢发育各期均有表达,在卵巢Ⅲ期表达量最高,其次是V期;在Cu~(2+)胁迫下,其表达量总体呈现先下降后上升然后又下降的趋势;在环境pH为9胁迫下,表达趋势呈现先下降,后回升,然后又下降,再大幅上升的趋势。研究表明,SeGPx在斑节对虾卵巢发育过程中具有重要作用,且Se-GPx参与斑节对虾对环境理化因子应激的免疫调控。     

植物遗传转化方法及其在棉花品质改良育种中的应用

农杆菌介导法、花粉管通道法和基因枪法是植物遗传转化的主要方法，这些转化方法在植物基因工程育种和基因改良中起到了重要作用。这些遗传转化方法目前已经广泛应用于棉花抗虫、抗病和品质改良育种实践，并获得了抗虫、抗病等性状良好的转基因植株，有些已经进入商品化生产，取得了显著的经济效益和社会效益。本文重点综述了这3种应用较广的遗传转化方法及其在棉花抗虫、抗病和品质改良育种中的应用，并讨论了我国转基因棉花研究的进展。     

咖啡营养特性及营养诊断研究进展

饮料作物咖啡（Coffea）为茜草科咖啡属的多年生灌木或者小乔木,在我国主要种植于云南与海南两省。本文概述了咖啡的主要营养特性及咖啡营养诊断方面的研究概况,并指出咖啡营养诊断的研究方向。     

Food ingestion,consumption and selectivity of pompano,Trachinotus ovatus (Linnaeus 1758) under different rotifer densities

The growth, survival, food selection and consumption of pompano larvae under different rotifer densities as well as their colour preference during the rotifer feeding stage were examined in this study. Growth and survival of fish larvae were not significantly affected when rotifer density was between 10 and 20 mL^?1. Fish larvae grew slower at 1 and 40 rotifers mL^?1 than at 10 and 20 rotifers mL^?1, and higher fish survival was achieved when fish larvae were exposed to 10 and 20 rotifers mL^?1. The rotifer density of 1 mL^?1 not only reduced food ingestion during the early stage, but also delayed diet switch from rotifer to copepod nauplii. On 5 days post hatching (DPH), larval pompano ingested more rotifers in dark‐coloured tanks and ingested more rotifers when prey colour was green. Based on the results obtained in the present study, the culture of larval pompano larvae is recommended using dark wall tanks with a feeding density of 10–20 rotifers mL^?1 during the initial feeding stage. This study proposes a management protocol to use appropriate type and quantity of live food to feed pompano larvae in a hatchery rearing condition, which could be applicable to the culture of fish larvae in other marine fish species.     

鲮微卫星引物的鉴定及其适用性检验

利用4种鲤科鱼类的14对微卫星引物对西江流域一批野生鲮进行PCR扩增。将各扩增条带进行克隆测序,发现引物MFW1、MFW2、MFW15、MFW17、Cc7、Cc11、Bgon22扩增出的产物含有微卫星重复序列。进一步对鲮的微卫星位点MFW1、MFW2、Cc7重新设计引物,并将其分别命名为Cm1、Cm2、Cm3,新引物对鲮的扩增特异性增强。采用引物Cm1、Cm2、Cm3、Cc11、MFW15、MFW17、Bgon22对西江流域一批野生鲮进行引物适用性检验。结果表明,除引物MFW15、Cc11无多态性外,其余5对引物(Cm1、Cm2、Cm3、MFW17、Bgon22)在取样群体中扩增图谱带型丰富,随引物不同,各标记在群体中检测到的等位基因数为2到16个。各微卫星座位的期望杂合度(He)及观察杂合度(Ho)范围分别为0~0.9038和0~1,平均分别为0.6881(0.1819SD)和0.7772(0.1931SD)。座位连锁分析显示Cm1与Bgon22之间存在显著性水平连锁关系(P<0.05),其余各座位之间未检测到明显的连锁关系(P>0.05)。研究群体的遗传多样性指数平均为0.6823,多态性水平相对较高。以上结果表明,筛选获得的7个微卫星座位适于对鲮进行遗传多样性分析。     

斑节对虾6个家系生长、饲料利用和全虾营养成分的比较

以海南三亚野生斑节对虾( Penaeus monodon )为亲本,建立6个家系(hsy5-5,hsy7-7,hsy8-5,hsy10-10,hsy12-13和hsy15-15)进行生长、饲料利用和混养生长试验,同时分析各个家系全虾的营养成分.在生长和饲料利用试验中,家系hsy10-10的增重率和饲料系数优于其它家系;hsy8-5、hsy10-10和hsy12-13的成活率高于hsy5-5、hsy7-7和hsy15-15,但差异不显著( P ＞0.05).在混养生长试验中,水泥池混养时,家系hsy10-10的成活率高达90%,分别比hsy8-5、hsy12-13、hsy7-7、hsy15-15和hsy5-5家系高出13.3%、13.3%、20.0%、23.3%和30.0%;hsy10-10和hsy8-5的月总增重率高于其它4个家系;池塘混养时,hsy10-10家系的平均体重高于6个家系平均值的13.1%.结果表明,家系hsy10-10的成活率、增重率和饲料系数均优于其它家系,且含有最高的粗蛋白、粗脂肪和最低的水分.     

紫茎泽兰提取物对棉蚜的毒力及其灭蚜机理研究

紫茎泽兰Eupatorium adenophorum提取物对棉蚜Aphis gossypii有很高的毒杀作用。紫茎泽兰乙醇总提物、其氯仿萃取物以及从氯仿萃取物中分离得到的紫茎泽兰素A对棉蚜无翅成蚜的致死中浓度(LC_(50)浸渍法)分别为167.9、224.7、362.8mg/L。试验表明,紫茎泽兰素A对棉蚜钠-钾-三磷酸腺苷酸酶(Na-K-ATPase)活性有明显的抑制作用。棉蚜经2.00mg/ml紫茎泽兰素A溶液处理12h后,其体内的Na-K-ATPase活性仅为对照组棉蚜Na-K-ATPase活性的21.60％。