柞蚕品种间对微粒子虫的抗性测定

一种昆虫对某一病原的感受性不仅存在着地域差异,同时还存在着品系及品种间的差异,不同蚕品种对微粒子病的感受性有强弱之分(三谷贤三郎,1930),在家蚕品种中抗性强的ID50值是抗性弱的100倍,这种感受性强弱可遗传给后代,认为在不同家蚕品种之中存在不同的微粒子病抗性基因.     

家蚕病毒病抗性研究进展

家蚕病毒病的发病率高、防治难度大、危害程度高。本文综述了近十年来在家蚕核型多角体病毒、质型多角体病毒、浓核病病毒和软化病病毒方面的抗性品种选育、抗性功能基因、抗性功能蛋白、表观遗传等方面的研究进展,对今后运用多手段进行家蚕病毒病防治和抗性理论研究提供参考,为抗病毒药物、抗病毒疫苗的研发提供新的思路。     

家蚕AN品系的核型多角体病毒(BmNPV)抗性基因连锁与定位分析

不同家蚕品种对家蚕核型多角体病毒(Bm NPV)的抵抗性存在较大差异。以对Bm NPV感染具有较强耐受性的家蚕品种华康2号杂交组合(F1)累代自交建立AN品系,以Bm NPV多角体悬液对该品系2龄起蚕经口添食的致死中浓度(LC_(50))为2.154×10~9m L~(-1),比对易感品系C108的LC_(50)提高了10~4倍以上。以易感品系C108作为回交亲本与AN品系组配F_1、BC_1、BC_2群体,2龄起蚕用浓度为1×10~8m L~(-1)的Bm NPV多角体悬液浸渍的桑叶饲喂12 h进行抗性遗传分析,表明AN品系对Bm NPV的高度抗性由位于常染色体的1个显性主基因控制。利用SSR分子标记和高分辨率熔解曲线(HRM)分析技术对BC2F群体进行连锁分析,发现AN品系的Bm NPV抗性主基因位于第27连锁群。依据对BC2M定位群体的600余个个体的HRM分析结果,绘制了遗传总距离为29.1 c M的AN品系抗性主基因遗传连锁图,抗性主基因与S2800-9和S2801-352标记的遗传距离分别为5.2 c M、2.9 c M。通过对添食中等浓度(1×10~7m L~(-1))Bm NPV多角体悬液后存活个体的基因组DNA进行SSR分子标记HRM分析,推测家蚕抗性品系AN还存在多个分布于其他染色体的微效抗性基因。     

野桑蚕与家蚕对敌敌畏的抗性研究初报

野桑蚕是家蚕的亲缘类群,其遗传背景、生活习性及对农药的抗性等方面与家蚕有一定的相关性。在蚕期,野桑蚕作为危害 桑树的主要害虫之一,对其进行农药防治十分困难。蚕期常用的农药是敌敌畏,本文就野桑蚕和家蚕对敌敌畏抗性进行了分别调查和研究。     

利用转基因RNA干涉提高家蚕对BmNPV的抗性初探

基于探讨利用转基因RNA干涉技术使家蚕获得对核型多角体病毒抗性的目的,将家蚕核型多角体病毒(BmNPV)的DNA聚合酶基因、蛋白激酶(PK1)基因以及bro-d和orf1629基因的部分编码片段,分别以其反向重复的形式与家蚕Actin3启动子连接,构建了基于piggyBac转座子载体的转基因表达载体,通过显微注射于家蚕卵,获得了36~107个G1蛾区,得到了20~23头转基因阳性家蚕。经Southern杂交及RT-PCR分析表明,BmNPV的4个反向重复片段都已经导入家蚕基因组中,并且可以进行转录。攻毒实验结果表明,BmNPV的DNA聚合酶基因和PK1基因反向重复片段对病毒的增殖有抑制作用,从而使家蚕对BmNPV具有一定抗性;而BmNPV的bro-d和orf1629基因反向重复片段对病毒的增殖没有抑制作用。针对特定病毒的转基因RNAi技术有望使家蚕获得对该病毒的抗性。     

家蚕核型多角体病毒及抗病育种

本文介绍了家蚕核型多角体病毒的发现、性状、复制机制以及对家蚕核型多角体病毒病的防治方法。不同的研究者用不同的材料研究了家蚕对NPV的抵抗性及其遗传规律,得到了不同的结果。我们的研究发现了高抗NPV的家蚕特优种质,探明了家蚕对BmNPV抗性的遗传规律。以306和NB为亲本,组配了近等基因系,筛选了与常染色体上抗NPV基因连锁的RAPD分子标记,并以NB为素材,育成了一批新种质。     

家蚕抗质型多角体病毒(BmCPV)的研究进展

由家蚕质型多角体病毒(Bombyx mori cytoplasmic polyhedosis virus,Bm CPV)感染引发的家蚕中肠型脓病对蚕业生产的危害严重,目前对该病害的防治仍局限于消毒预防。国内外学者希望从筛选的对Bm CPV有较强抵抗力的家蚕品种中寻找抗性基因和分析抗性机制,并通过转基因技术过量表达抗性基因或经RNA干扰病毒侵染增殖关键基因等策略来提高家蚕防御Bm CPV侵染的能力。本文综述家蚕抗Bm CPV研究在上述方面取得的重要进展,并对家蚕抗Bm CPV的分子机制进行分析与讨论,为该领域展开更深入的研究提供较为系统的参考信息。     

家蚕脂肪酶基因Bmlipase-1在不同BmNPV抗性水平家蚕品种间的表达差异

家蚕脂肪酶Bmlipase-1在家蚕中肠组织特异性表达,具有抵抗家蚕核型多角体病毒(BmNPV)感染的活性。以对BmNPV具有不同抗性水平的6个家蚕品种为材料,利用荧光定量PCR(qRT-PCR)技术,检测Bmlipase-1基因在不同家蚕品种5龄期健康幼虫中肠组织的表达水平及感染BmNPV后的5龄期幼虫中肠组织中该基因的表达水平变化,探究该基因表达水平与家蚕对BmNPV的抗性水平的关系。结果表明:不同家蚕品种间Bmlipase-1的表达水平差异显著,抗性较强的品种,其表达水平较高,6个供试品种5龄幼虫中肠组织中的Bmlipase-1表达水平依次为NIL.LVR>P50>CVDAR18>nsd.NIL>892>306,抗性较强品种NIL.LVR 5龄期的平均表达量约为抗性较弱品种306的8.2倍;BmNPV能够诱导Bmlipase-1的表达,但不同品种间该基因的诱导表达差异显著,仍表现为抗性较强的品种该基因的诱导表达水平较高,NIL.LVR经BmNPV感染诱导后Bmlipase-1在5龄期的平均表达量约为抗性较弱品种306平均诱导表达水平的12.4倍。推测Bmlipase-1的表达水平与蚕体自身对BmNPV的抗性水平有一定的关联性。     

数量遗传学在家蚕育种上的应用概述

通过查阅家蚕经济性状遗传研究文献,概括叙述了数量遗传学在家蚕数量性状常规研究、品种稳定性分析、抗性遗传研究、饲料效率遗传研究及QTLs研究等方面的应用情况及部分成果.     

家蚕对丙体—666抵抗性及其遗传研究

用点滴法测定家蚕对丙体—666(γ—BHC)的抵抗性,探讨了抗性与发育时期、生理状况的关系,抗性与茧质、体质的关系,抗性的遗传。结果表明:雄蚕抗性略大于雌蚕;随着蚕的发育,抵抗性也增大。但以单位体重的抗性值,分析同一品种不同龄期几乎无差异;起蚕食桑后抗性显著增强,其增强程度超过体重增长率;抵抗性与体重、全茧量、茧层量、茧层率均有正的相关性,与死笼率无相关性;品种间抵抗性的LD50值(微克/克)的对数值近似正态分布;家蚕对γ—BHC的抵抗性受多基因控制,无母体影响。但由新九与兰5一组杂交试验的分析,认为还有不完全显性的主基因存在,位于常染色体上。     

柞蚕抗病机制及其抗病物质的研究——Ⅲ.柞蚕诱导抗菌物质活性与柞蚕对NPV感染抵抗性的相关关系

本试验利用半蛾区测定方法,将同蛾区蛹期诱导抗菌物质活性、稚蚕期对柞蚕NPV的感染抵抗性进行了相关性的比较试验.初步结果表明:(1)同蛾区稚蚕期对NPV感染抵抗性强的.蛹期诱导抗菌物质活性也高,呈正相关关系,(2)蛹期诱导抗菌物质活性高的峨区,次代稚蚕期抗毒力(NPV)也强,呈正相关;(3)柞蚕五龄幼虫经大肠杆菌诱导后也能产生抗菌物质,且其诱导抗菌物质活性与溶菌酶活性趋于正相关.同时,供试的三个柞蚕品种,蚕期和蛹期诱导抗菌物质活性趋于一致.     

家蚕现行品种对氟化物、核型多角体病毒病抵抗性的研究

对若干家蚕现行新品种进行抗氟性能和抗核型多角体病毒（ＶＰＶ）病鉴定，结果是：不同品种对氟化物和ＶＰＶ病抵抗力有差异，抗氟能力较强的有丰一、８７２、５４Ａ、丰一×５４Ａ、华峰×雪松、８７１×８７２：抗ＮＰＶ病较强的是３１８、４１６、秋３、３１７×３１８、４１５×４１６、８７１×８７２；抗氟性和抗ＮＰＶ病能力没有完全一致性，但３１７×３１８、８７１×８７２综合抗性较强。杂交种的抗氟和抗ＮＰＶ病明显强于纯系。     

两广二号一代杂交种对NPV的抗性研究

家蚕血液性脓病是夏季养蚕极难控制的毁灭性蚕病,通过开展两广二号一代杂交种对NPV的抗性研究试验,得出结论:两广二号品种对NPV多角体病原具有很强的抗性,是陕南地区当前最适合饲养的夏用品种。     

影响家蚕中肠碱性磷酸酶活性的若干因素

研究了影响家蚕中肠碱性磷酸酶活性的若干因素,在供试的蚕品种间,酶的活性差异显著,且与茧质成绩呈正相关;NPV与CPV对中肠碱性磷酸酶活性的影响不同,即CPV感染明显降低了此酶的活性,而NPV则无其影响;苏云金杆菌感染直接破坏中肠组织,引起酶活性的陡降;各种化学物质中以Mg2+对中肠碱性磷酸酶的激活作用最明显,体外条件下维生素C,Ca2-,柠檬酸,Cl-对酶有抑制作用。     

苜蓿银纹夜蛾核多角体病毒在家蚕蛹体内的复制及重组病毒外源基因的表达

家蚕蛹在复眼着色期,经4℃冷藏24小时后,可被Ac NPV(苜蓿银纹夜蛾核多角体病毒)感染。在蛹体内复制出的多角体大小差异较大,且比在昆虫Sf—21细胞中繁殖的Ac NPV和在蚕蛹体内形成的Bm NPV多角体小。在蛹体内繁殖的Ac NPV的游离病毒可以回返感染Sf—21细胞。由蚕蛹内分离的Ac NPV基因组DNA的限制性内切酶图谱与野生型的Ac NPV相同。以上结果证明Ac NPV可以在蚕蛹体内复制。含HBsAg基因(人乙肝表面抗原)的重组Ac NPV亦可感染家蚕蛹,并能正确表达外源基因。此外,还发现化蛹后第2天的家蚕蛹被注射约5×10~5PFU的Ac NPV,可诱导“人工滞育蛹”现象的发生,在25℃经过30天后蛹仍存活,发育停滞在复眼着色前阶段。     

家蚕核多角体病毒DNA聚合酶基因的克隆

从家蚕核多角体病毒（ＢｍＮＰＶ）基因组中克隆了完整的ＢｍＮＰＶＤＮＡ聚合酶基因。详细的限制性内切酶酶谱分析表明：ＢｍＮＰＶＤＮＡ聚合酶基因的限制性内切酶图谱与苜蓿尺蠖核多角体病毒（ＡｃＭ－ＮＰＶ）ＤＮＡ的聚合酶基因图谱完全一致，且在基因组中的位置亦相似。ＤＮＡ聚合酶基因的部分测序结果显示：ＢｍＮＰＶ与ＡｃＭＮＰＶ的ＤＮＡ聚合酶有高度的同源性，远高于ＡｃＭＮＰＶ和舞毒蛾核多角体病毒（ＬｄＭＮＰＶ）两者的ＤＮＡ聚合酶间的同源性，说明ＡｃＭＮＰＶ和ＢｍＮＰＶ间的亲缘关系比ＬｄＭＮＰＶ要近，从分子进化的角度来看，用表型性状进行杆状病毒科属以下的分类似乎并不适宜。     

EFFECTS OF OBSERVER ON THE DIAGNOSTIC ACCURACY OF LOW‐FIELD MRI FOR DETECTING CANINE MENISCAL TEARS

Low‐field MRI (lfMRI) has become increasingly accepted as a method for diagnosing canine meniscal tears in clinical practice. However, observer effects on diagnostic accuracy have not been previously reported. In this study, 50 consecutive stifle joints with clinical and radiologic evidence of cranial cruciate ligament insufficiency were investigated by lfMRI and arthroscopy. Fifteen observers who had varying levels of experience and who were unaware of arthroscopic findings independently reviewed lfMRI studies and recorded whether lateral and medial meniscal tears were present. Diagnostic accuracy (sensitivity, specificity, positive (PPV) and negative predictive value (NPV)) was determined for each observer and median values were calculated for all observers, using arthroscopy as the reference standard. Interrater agreement was determined based on intraclass correlation coefficient (ICC) analysis. Observer level of experience was compared with diagnostic sensitivity and specificity using correlation analysis. Based on pooled data for all observers, median sensitivity, specificity, PPV, and NPV for lfMRI diagnosis of lateral meniscal tears were 0.00, 0.94, 0.05, and 0.94, respectively. Median sensitivity, specificity, PPV, and NPV for medial meniscal tears were 0.74, 0.89, 0.83, and 0.79, respectively. Interrater agreement for all menisci was fair (0.51). Menisci were less consistently scored as having no tears (ICC = 0.13) than those scored as having tears (ICC = 0.50). No significant correlations between observer experience and diagnostic sensitivity/specificity were identified. Findings indicated that the accuracy of lfMRI for diagnosing canine meniscal tears was poor to fair and observer‐dependent. Future studies are needed to develop standardized and widely accepted lfMRI criteria for diagnosing meniscal tears.     

THE POTENTIAL OF SOME SYNTHETIC PYRETHROIDS FOR CONTROL OF THE CATTLE TICK (BOOPHILUS MICROPLUS)

As part of a continuing programme to aid in the development of new acaricides, the potentials of three synthetic pyrethroids, permethrin, cypermethrin and decamethrin, were assessed against a range of resistant strains of the cattle tick (Boophilus microplus). The compounds were shown to be acaricidal, with decamethrin the most effective, giving efficient control of susceptible and organo-phosphorus resistant strains of the cattle tick at concentrations as low as one fifth of those previously used for any commercial acaricide. However higher concentrations were required for satisfactory control of the DDT-resistant strain. Permethrin-selection of a field strain containing a low percentage of DDT-resistant ticks led to rapid development of resistance to a level equivalent to that of the DDT-resistant strain. Further selection increased resistance to permethrin to a higher level than that shown in the DDT-resistant strain. This increased resistance to permethrin appears to be restricted mainly to this compound and is not associated with a marked increase in resistance to cypermethrin, decamethrin or DDT. The potential of organo-phosphorus compounds to potentiate the pyrethroids when the two classes of compounds are used as mixtures against the pyrethroid-resistant and susceptible strains was confirmed.     

Bioeconomic evaluation of embryo transfer in beef production systems: III. Embryo lines for producing bulls.

A model was developed for the economic evaluation of embryos for producing bull lines for use in commercial beef production. The fundamental concept underlying the model is that a cloned and sexed embryo of known genetic characteristics for beef traits is used to produce a bull. After reaching physiological maturity, the bull is used in natural matings. Equations relating feed energy requirements and growth rates based on NRC requirements and costs and returns discounted to present value allow investigation of expected economic merits of progeny from different embryo bull lines. The model has the flexibility to determine optimal embryo characteristics for different production environments. Model sensitivity to variation in progeny sex ratios, growth rates, yield and quality grades, and herd fertility characteristics was examined. Net present values (NPV) per embryo transferred were determined at the optimal marketing age of progeny produced from mating the bull to 30 cows per year for 5 yr. Relative to the lowest NPV of $18,209 for progeny with an expected quality grade of Select and yield grade of 4 at 400 d, increments in NPV ranged from $329 to $22,708 depending on differences in expected progeny carcass grade characteristics. The difference between NPV for 100% male and 40% male sex ratios was $7,518. The NPV differences between progeny growth rates of 1.6 and .9 kg/d holding herd conception rate constant at .9 and .5 were $8,311 and $4,611, respectively. The model evaluates relative economic values of embryo lines for producing bulls, accommodating interactions among progeny characteristics, and environments.     

野桑蚕NPV与家蚕NPV的外形差异及多角体蛋白基因的研究

通过电子显微镜观察 ,发现从野桑蚕体内分离的核型多角体病毒 (BmmNPV)经空斑筛选 ,其分离株的多角体外形和家蚕核型多角体病毒 (BmNPV)的多角体外形呈现差异。对PCR扩增出的BmmNPV的多角体蛋白基因polh进行克隆、测序 ,并与BmNPV的polh序列比较 ,结果发现BmmNPV的polh在 +2 2、+70 3和 +711的 3个位点的碱基不同 ,使 +7位、+2 99位的Pro变成了Ser,+711位G变成为A ,而没有引起该位点 +2 31位Ala的改变。由Ser和Pro在形成蛋白二级结构的构象常数可以推测 ,BmmNPV的多角体较BmNPV更稳定 ,查明了多角体在外形存在差异的原因是ORF内碱基的差异引起的。