茶刺蛾核型多角体病毒实用剂型的配制

通过使用剂量的生物测定,明确了茶刺蛾核型多角体病毒(IfNPV)制剂中病毒的合适含量为1×107PIB/ml～2×107PIB/ml.测定了IfNPV(1×107PIB/ml)和Bt(2000 IU/μl)混配后的联合毒力,结果表明,协同毒力指数为2.1%,两者为相加作用.在此基础上,配制了茶刺蛾病毒水剂和茶刺蛾病毒Bt混剂,两种制剂的室内外毒效均在99%以上.     

茶刺蛾核型多角体病毒制剂示范应用试验

茶刺蛾（Iragoides fasciata Moore）是我国茶树上的一种重要害虫，爆发成灾时，可将成片茶园食成秃枝，严重影响产量。茶刺蛾幼虫的毒刺，也严重影响茶叶采摘和田间管理。近年来，茶刺蛾在有机茶园为害猖撅．为寻找有效防治茶刺蛾的生物制剂，笔者对茶刺蛾核型多角体病毒（IfNPV）制剂进行了研究，现将其示范应用试验结果报道如下。     

灰茶尺蛾核型多角体病毒的初步研究

1987年在武昌县茶园中采集的灰茶尺蛾(Ecxtropis grisescens Warren)死虫标本,分离出一种核型多角体病毒。通过对该病毒的形态结构、病症及侵染虫体的组织部位的观察,发现属一种新的杆状核型多角体病毒,其分离观察结果如下: 将茶园中采集到的病死虫用0.1摩PBS     

显脉球须刺蛾多角体病毒的初步研究

从显脉球须刺蛾(Scopelodes venosa Kwangtungensis Hering)幼虫自然罹病死亡虫体中,分离到1株多角体病毒,包涵体呈多角形,有四边、六边、八边等多种形态,大小为600—800×1000—1200nm,多角体会在0.05 mol/L Na_2CO_3+0.1 mol/L NaCl 弱碱溶液中降解,且能释放出短杆状的病毒粒子,其大小为260—300×40—60 nm。室内感染3—4龄幼虫,LC_(50)为4.112×10~3 PIB,回归直线方程为 y=0.549x+2.86     

茶尺蠖核型多角体病毒制剂的防治效果

茶尺蠖核型多角体病毒(Ectropis obliqua nuclear polyhedrosis virus,简称 EONPV)的两种制剂 EONPV 2H_2 的 Lc_(50)为2.5×10~4 PIB/m1、EONPV 2H_4的Lc_(50)为2×10~4 PIB/ml,以1×10~7 PIB/ml的 EONPV 2H_2感染寄主3龄虫的 Lt_(50)为5.5天,EONPV 2H_4 的 Lt_(50)为5.7天。两制剂均有一定的防紫外光能力,对寄主的致病性强,室内试验校正死亡率都在96%以上,     

茶尺Huo核型多角体病毒与农药混用试验

本文测定了茶尺Ｈｕｏ核型多角体病毒与几种杀虫剂混用的杀虫效果。     

茶尺蠖核型多角体病毒的生产工艺流程

茶尺蠖核型多角体病毒毒力较强,适宜于作为杀虫剂用于茶尺蠖防治。但要在生产上推广应用,首先必须解决工厂化大量生产问题。因此,我们在饲料筛选、病毒增殖条件等研究基础上,对该病毒的生产工艺流程进行了探索。现将主要流程概括如下。一、健虫饲养1.健虫室及养虫器具消毒健虫室在每次使用前5～6天,用蚕室蚕具消毒剂——消特灵(杭州江南植物生长素厂生产)澄清液喷布消毒,并打开室内紫外光灯照射2小时。养虫用500g 装玻璃果酱瓶、2×9cm 平底玻管,置120～150℃下高温干燥消毒1小时,其他木结构养虫器具用消特     

茶尺蠖核型多角体病毒与农药混用试验

本文测定了茶尺蠖核型多角体病毒（EoNPV）与几种杀虫剂混用的杀虫效果。生物测定结果表明,苏云金杆菌（Bt）与EoNPV混用时,表现有一定的颉颃作用,而且Bt浓度越高,往往颉颃作用越强。敌灭灵、喹硫磷与EoNPV混用,虽然有协同作用,但无增效作用。     

日龄对茶尺蠖核型多角体病毒毒性的反应

室内生测表明 ,1～ 6日龄的茶尺蠖幼虫感染 Eo NPV 9～ 10天后均 10 0 %死亡 ,日龄对最终毒性无影响。但不同日龄幼虫的起始死亡时间和 L T50 有明显差异。 L T50 为 4 .3± 0 .2 0天～ 6 .7± 0 .14天 ,各日龄虫的 L T50按从低到高排列次序为 :1日龄 <5日龄 <4日龄 <2日龄 <3日龄 <6日龄。     

灰茶尺蠖核型多角体病毒(EgNPV)安全性试验

用20亿PIB/mL灰茶尺蠖病毒(EgNPV)对成年SD大鼠进行急性经口、经皮毒性试验,对白色家兔进行眼刺激和皮肤刺激试验,对豚鼠进行皮肤致敏试验以及进行小鼠急性致病性试验。结果表明20亿PIB/mL灰茶尺蠖病毒(EgNPV)水剂对SD大鼠的急性经口毒性(其LD50雌雄大鼠均5 000 mg/kg)属于低毒;对SD大鼠的急性经皮毒性(其LD50雌雄大鼠均2 000 mg/kg)属于低毒;对家兔的皮肤无刺激性;对家兔的眼睛也无刺激性:在染毒后24 h内不洗眼情况下,眼刺激积分最高为0;对豚鼠的皮肤致敏率为0,其致敏性分级为Ⅰ级,属弱致敏物;病理组织学检查结果表明,各实验组小鼠内脏组织均未见有病理改变,无急性致病性。     

玉米螟虫生真菌球孢白僵菌几丁酶基因的克隆与分析

球孢白僵菌是最常见的昆虫病原真菌之一,已成功应用于玉米螟和松毛虫等害虫的生物防治。球孢白僵菌主要通过分泌多种胞外水解酶降解昆虫体壁侵入寄主,其分泌的水解酶主要包括蛋白酶、几丁质酶、脂酶等。本研究从玉米螟虫生真菌球孢白僵菌中分离到一个几丁质酶基因BbChitI,该基因的编码区包含1047bp,编码了348个氨基酸,氨基酸序列与来源于小菜蛾虫生真菌的白僵菌几丁质酶的同源性高达99%,与来源于木霉菌的几丁质酶的同源性达到82%。     

茶树亲环素基因cDNA全长的分析鉴定与原核表达

通过对茶树新梢cDNA文库大量随机测序,获得了一个编码亲环素的全长基因,在GenBank登录,登录号为DQ904327。茶树亲环素基因cDNA全长949 bp,其中开放阅读框全长495 bp,编码蛋白质含有164个氨基酸,分子量约为17.47 kD,等电点约为8.54,它具备5’端非编码区的“CAAT”标志及3’端非编码区的”AATAAA”poly-A加尾信号。其推测的氨基酸序列经与26条其他生物亲环素蛋白氨基酸序列进行CLUSTAL W多序列联配并以Neighbor-Joining法进行进化树构建后,发现与水稻和小麦的相似性较高,达到85%以上。根据亲环素基因开放阅读框序列设计引物,构建了原核表达载体pET/Csin-Cyp,并在大肠杆菌BL21(DE3)中成功诱导出了一个分子量为23 kD的亲环素融合蛋白。     

茶树硝酸根转运蛋白基因NRT2.5的克隆及表达分析

通过RACE克隆从茶树[Camellia sinensis (L.)]叶片中获得NRT2.5基因cDNA全长序列。所得基因序列全长为2 457 bp,其中开放阅读框为1 362 bp,编码454个氨基酸,推测蛋白分子量为48.7 kD,理论等电点为9.63。生物信息学分析表明,该基因与可可树、拟南芥等的NRT2.5基因具有高度同源性,且其编码的蛋白质具有NRT家族共有的结构特征。实时定量PCR分析表明,NRT2.5基因在茶树不同组织中均有表达,但主要在成熟叶和根中表达;不同氮浓度处理后,茶树NRT2.5基因在低浓度下的表达量高于高浓度下。     

茶树ACC合成酶基因全长cDNA的克隆及其生物信息学分析

乙烯是一种重要的植物激素,参与植物许多生理过程。ACC(1-氨基环丙烷-1-羧酸)合成酶(ACS)是植物乙烯合成过程中的限速酶,对乙烯的合成具有重要的调控作用。利用其它植物ACS基因的保守序列设计兼并引物,采用RACE(Rapid Amplificationc DNA Ends)和RT-PCR等技术,克隆得到编码茶树ACS基因的全长cDNA序列,长1579bp,编码478个氨基酸,预测分子量约为53.7kD,等电点8.039,GenBank登录为EF205149。以Neighbor-Joining法构建进化树,发现茶树ACS基因与柿树中的同类基因亲缘关系最近。茶树ACS基因的克隆为从分子水平认识乙烯在茶树上的生理作用奠定了基础。     

番木瓜镁离子转运蛋白基因CpMGT1的克隆与功能分析

镁是一种大量金属营养元素,其在植物的生长、发育、光合、胁迫响应等生物学过程中起重要作用。在高等植物中,Mg²⁺的吸收、运输、分布和再分配主要由镁离子转运蛋白（MGT/MRS2）和Mg²⁺/H⁺交换体（MHX）介导。其中,MGT/MRS2又名CorA,最先在鼠伤寒沙门氏菌中被发现,后在拟南芥、水稻等模式植物中进行了较为深入的研究。番木瓜（Carica papaya L.）是一种隶属于十字花目番木瓜科的重要热带果树,至今还未见有关MGT基因的报道。本研究基于番木瓜的基因组和转录组数据,采用RT-PCR技术成功克隆到一个MGT基因（CpMGT1）,应用生物信息学手段预测蛋白的理化特性和保守基序,运用qRT-PCR分析基因的表达模式,利用缺失CorA、MgtA和MgtB的沙门氏菌突变株MM281进行功能互补。结果表明：CpMGT1的编码区为1332 bp,预测编码443 aa,理论分子量为50.43 kDa、等电点为5.12;该蛋白含有2个疏水跨膜区（TM1和TM2）,其中TM1包含高度保守的GMN基序;进化及亚细胞定位分析表明CpMGT1与拟南芥中AtMGT1和AtMGT2的亲缘关系较近,定位于细胞膜;定量分析显示CpMGT1基因为组成型表达,其中在根、茎和果实中的表达量较高;在MM281中异源表达可显著提高工程菌在低Mg²⁺浓度下的生长速率,表明CpMGT1具有高效的Mg²⁺转运活性。CpMGT1的克隆与鉴定为进一步揭示番木瓜MGT基因在不同组织特别是在果实中Mg²⁺的积累机制奠定了坚实的基础。     

The Antinociceptive and Anti-Inflammatory Activities of Caulerpin,a Bisindole Alkaloid Isolated from Seaweeds of the Genus Caulerpa

The antinociceptive and anti-inflammatory activity of caulerpin was investigated. This bisindole alkaloid was isolated from the lipoid extract of Caulerpa racemosa and its structure was identified by spectroscopic methods, including IR and NMR techniques. The pharmacological assays used were the writhing and the hot plate tests, the formalin-induced pain, the capsaicin-induced ear edema and the carrageenan-induced peritonitis. Caulerpin was given orally at a concentration of 100 μmol/kg. In the abdominal constriction test caulerpin showed reduction in the acetic acid-induced nociception at 0.0945 μmol (0.0103–1.0984) and for dypirone it was 0.0426 μmol (0.0092–0.1972). In the hot plate test in vivo the inhibition of nociception by caulerpin (100 μmol/kg, p.o.) was also favorable. This result suggests that this compound exhibits a central activity, without changing the motor activity (seen in the rotarod test). Caulerpin (100 μmol/kg, p.o.) reduced the formalin effects in both phases by 35.4% and 45.6%, respectively. The possible anti-inflammatory activity observed in the second phase in the formalin test of caulerpin (100 μmol/kg, p.o.) was confirmed on the capsaicin-induced ear edema model, where an inhibition of 55.8% was presented. Indeed, it was also observed in the carrageenan-induced peritonitis that caulerpin (100 μmol/kg, p.o.) exhibited anti-inflammatory activity, reducing significantly the number of recruit cells by 48.3%. Pharmacological studies are continuing in order to characterize the mechanism(s) responsible for the antinociceptive and anti-inflammatory actions and also to identify other active principles present in Caulerpa racemosa.     

Cytotoxicity and Inhibition of Lymphocyte Proliferation of Fasciculatin,a Linear Furanosesterterpene Isolated from Ircinia variabilis Collected from the Atlantic Coast of Morocco

Fasciculatin, a furanosesterterpene isolated from the marine sponge Ircinia variabilis from the Atlantic Coast of Morocco, has been evaluated for its influence on a mitogen-induced proliferation of human lymphocytes and growth of human tumor cell lines.     

青天葵独脚金内酯信号转导关键基因D14的克隆与亚细胞定位分析

D14基因是独脚金内酯信号转导途径的关键基因,其编码的蛋白能够使SLs释放出活性小分子物质而发挥调节腋芽起始的功能。本研究根据其他物种的D14基因从青天葵转录组数据中筛选获得2条高度同源的片段,命名为NfD14a和NfD14b。应用RT-PCR和RACE技术,克隆获得NfD14a和NfD14b的cDNA全长序列,GenBank登录号分别为MH028026、MH028027。NfD14a基因全长1206 bp,ORF为861 bp,编码286个氨基酸;NfD14b基因全长1082 bp,ORF为813 bp,编码270个氨基酸。对NfD14的编码蛋白序列进行了生物信息学分析,结果表明：NfD14a和NfD14b均属于α/β折叠蛋白水解酶超家族成员（Abhydrolase superfamily）,但系统进化分析结果表明两者同源性不高。利用一步快速克隆的方法构建了植物表达载体35S::NfD14a-EGFP和35S::NfD14b-EGFP,并分别获得其工程菌。瞬时表达结果表明,NfD14a和NfD14b均定位在烟草原生质体的细胞核和细胞质。本研究通过对NfD14基因全长cDNA序列的克隆与亚细胞定位分析,为青天葵独脚金内酯信号转导途径调控植物分枝生长发育机制的研究奠定基础。     

Synthesis and Antifouling Activity Evaluation of Analogs of Bromosphaerol,a Brominated Diterpene Isolated from the Red Alga Sphaerococcus coronopifolius

Marine biofouling is an epibiotic biological process that affects almost any kind of submerged surface, causing globally significant economic problems mainly for the shipping industry and aquaculture companies, and its prevention so far has been associated with adverse environmental effects for non-target organisms. Previously, we have identified bromosphaerol (1), a brominated diterpene isolated from the red alga Sphaerococcus coronopifolius, as a promising agent with significant antifouling activity, exerting strong anti-settlement activity against larvae of Amphibalanus (Balanus) amphitrite and very low toxicity. The significant antifouling activity and low toxicity of bromosphaerol (1) motivated us to explore its chemistry, aiming to optimize its antifouling potential through the preparation of a number of analogs. Following different synthetic routes, we successfully synthesized 15 structural analogs (2–16) of bromosphaerol (1), decorated with different functional groups. The anti-settlement activity (EC₅₀) and the degree of toxicity (LC₅₀) of the bromosphaerol derivatives were evaluated using cyprids and nauplii of the cirriped crustacean A. amphitrite as a model organism. Derivatives 2, 4, and 6–16 showed diverse levels of antifouling activity. Among them, compounds 9 and 13 can be considered as well-performing antifoulants, exerting their activity through a non-toxic mechanism.     

Gene Cloning,Expression and Characterization of a Novel Xylanase from the Marine Bacterium,Glaciecola mesophila KMM241

Marine xylanases are rather less studied compared to terrestrial xylanases. In this study, a new xylanase gene, xynB, was cloned from the marine bacterium, Glaciecola mesophila KMM241, and expressed in Escherichia coli. xynB encodes a multi-domain xylanase XynB of glycoside hydrolase (GH) family 8. The recombinant XynB comprises an N-terminal domain (NTD) with unknown function and a catalytic domain, which is structurally novel among the characterized xylanases of GH family 8. XynB has the highest identity (38%) to rXyn8 among the characterized xylanases. The recombinant XynB showed maximal activity at pH 6–7 and 35 °C. It is thermolabile and salt-tolerant. XynB is an endo-xylanase that demands at least five sugar moieties for effective cleavage and to hydrolyze xylohexaose and xylopentaose into xylotetraose, xylotriose and xylobiose. NTD was expressed in Escherichia coli to analyze its function. The recombinant NTD exhibited a high binding ability to insoluble xylan and avicel and little binding ability to chitosan and chitin. Since the NTD shows no obvious homology to any known carbohydrate-binding module (CBM) sequence in public databases, XynB may contain a new type of CBM.