The Study on Molecular Phylogenetic and Molecular Marker of Species in Silk Insects

本文对家蚕、野桑蚕、蓖麻蚕、柞蚕和天蚕等 5种绢丝昆虫进行了随机扩增多态DNA(RAPD)分析。 40个引物中有 2 7个引物能扩增出 5 36个清晰且重复性强的条带 ,其中可变条带数为 5 2 0个 ,单个引物扩增的条带数在11～ 2 8之间 ,平均为 19 9,各片段分子量大小在 0 2 9～ 2 6 7kb之间。每个样本都能找出其独特的分子标记。家蚕与野桑蚕的遗传距离 (D)最小 ,为 0 376 0 ;家蚕与蓖麻蚕的遗传距离 (D)最大 ,为 0 7488。根据遗传距离 ,用UPG MA聚类分析方法构建了它们的分子树。Five species of silk insects including Bombyx mori, B. manolarina, Philosamia cynthia, Autheraea pernyi and A. yamamai were analyzed by RAPD method using 40 arbitrary primers. In these primers, 27 of them could amplify clear and repeating bands. 536 fragments were obtained and the variable bands were 520. Each primer gave 11～28 bands and the average was 19.9. The length of the fragments is 0.29～2.67 kb. Some distinctive bands were found in every species. The genetic distance( D ) between bombyx mori and B. manolarina is 0.3760, which is the lowest. The highest D value is 0.7488, which between Bombyx mori and Philosamia cynhia. The D value was then used to construct a dendrogram by unweighted pair-group method with arithmetical averages(UPGMA).国家自然科学基金!资助项目 (批准号 39870 410

家蚕人工授精与精子超低温保存研究(简报)

家蚕人工授精与精子超低温保存研究（简报）桂慕燕陈元霖刘治纬吴维光（厦门大学细胞生物学研究室，厦门361005）（华南农业大学亚太地区蚕桑培训中心，广州510642）家蚕（bOMbyXMOrIl.）是一种很重要的经济昆虫，也是优良的实验生物。家蚕的人工...国家自然科学基

RAPD Analysis of Hereditary and Variation of Domesticated Silkworm Generated by Introduction of Eri Silkworm DNA

借助于精子介导，在家蚕受精的过程中将蓖麻蚕dnA转入家蚕卵内，从它们后代获得了新的变异品系。本文采用rAPd技术对这些品系基因组dnA进行了分析。结果表明，所用50种10MEr随机引物中有49个检测出dnA的多态性，统计分析图谱中各类扩增带，其中变异品系与其相应受体的差异带占其总带数的26~37%，提示外源dnA导入受体后引起后代基因组的显著变异，并对这些变异的意义进行了讨论。With the aid of domesticated silkworm sperms, eri silkworm DNA was transFerred into domesticated silkworm eggs during insemination, and variant strains were obtained From the progenies.Genomes of three new strains were analyzed using RAPD assay.Polymorphic Fingerprints were obtained From 49 out of 50 primers.DiFFerent kinds of ampliFied bands in RAPD patterns were calculated and analyzed, the variant bands between variants and their recipients counted For 26～37% of the total bands of each variant.The results indicated that exogenous DNA introduced into recipients induced remarkable variation in progeny genomes.The signiFicance of the variation was discussed.国家自然科学基

Application of RAPD Technique in Genetic Relationship Study of Silk Insect Ⅰ. Genetic Variance in Eri Silkworm

用RAPD技术对蓖麻蚕基因组DNA进行多态性研究，分析了5个蓖麻蚕品种间的遗传差异。结果表明，所采用的40个随机引物中，有27个引物扩增谱带清晰且重复性较好，扩增总片段数达243个，单个引物的扩增片段数在4～17之间，平均为9条，片段大小在033～30kb之间。不同蓖麻蚕品种间的遗传距离(Ｄ)在00683～01603之间，根据Ｄ值，由UPGMA聚类分析软件绘制了它们的聚类分子树。Random amplified Polymorphic DNA (RAPD) was used to analyze the genetic diversity among eri sickworm. The genetic variance of five erisickworm was studied. The result showed that: 27 of 40 arbitrary primers could amplify clearly with repeatable bands.243 fragments were obtained.Each primer gave 4～17 bands and the average was 9.The length of the band was 0.33～ 3.0kb. The genetic distance (Ｄ) value between different breeds of Eri Silkworm was 0.0683～0.1603. The D value was used to construct a dendrogram by UPGMA.国家自然科学基金资助项目!（批准号39870410

绢丝昆虫线粒体DNA多态性研究

采用差速离心法和碱变性法制备家蚕、野桑蚕、蓖麻蚕、柞蚕和天蚕蛹线粒体dnA（MTdnA），并进行了限制性酶切片段多态性（rflP）检测。结果表明，绢丝昆虫在种间MTdnA酶切类型差异明显，而在种内不同品种之间则没有差异或差异甚少。在HAEⅢ酶解作用下，家蚕的15个一化和二化性品种的酶切类型相同，5个多化性品种的酶切类型也相同，但它们彼此间则有差异，前者与野桑蚕属同一个类型。由此提示，在家蚕品种演化过程中，一化和二化品种可能与野桑蚕有更密切的亲缘关系

Studies on the Mechanism of Variations of Hybrids of Domesticated Silkworm and Eri Silkworm ——RAPD Analysis of Genome

采用24种随机引物，对以蓖麻蚕精子进行人工授精得到的家蚕后代中的3个稳定变异品系及其亲本的基因组进行了rAPd检测，结果显示，在变异品系的rAPd图谱中，不仅存在大量与母本相同的“亲本带”，同时还出现了不同数量与母本不同的“变异带”，包括“非亲本带”、“缺失带”及个别仅与父本相同的“目的带”，从分子水平上揭示了变异品系存在着明显的“偏母性”与“变异性”特点。Twenty-Four random primers were used to analyze the genomes of three descendant strains with steady hereditable variation produced Form domesticated silkworm by artiFicial insemination with sperms of eri silkworm.The results show that in the RAPD patterns there are many ampliFied bands called“ parental bands”which are similar to those of the Female parent.At the same time, there appears varied amount of ampliFied bands called “variant bands” that are diFFerent From those of the Female parent.The variant bands include non-parental-bands, lost-bands and several “expected-bands” which only shared with the male parent.This research reveals the signiFicant matrocliny and variation in the descendant strains at the molecular level.国家自然科学基

Comparison of four promoters for transient expression of RFP reporter gene in cultured Bombyx mori cells (Bm-e-HNU5)

以红色荧光蛋白基因(RFP)为报告基因,构建含4种不同启动子的重组表达质粒,用脂质体介导法转染家蚕Bombyxmori细胞(Bm-e-HNU5),观察家蚕细胞质肌动蛋白4基因启动子(A4)、α微管蛋白基因启动子(α-tub)、蚕丝心蛋白重链基因启动子(Fib)和家蚕核型多角体病毒早期即刻蛋白基因启动子(IE)4种启动子调控RFP报告基因在家蚕细胞内的瞬时表达情况。构建的重组表达质粒pDsRed-α-tub、pDsRed-A4、pDsRed-IE和pDsRed-Fib经双酶切和PCR鉴定正确无误。转染和转录实验结果表明,除了pDsRed-A4外,其他3种重组质粒在Bm-e-HNU5细胞中都得到高转染率,α-tub、IE和Fib可依次增强RFP报告基因在家蚕细胞内的瞬时表达活性。The red fluorescent protein reporter gene (RFP) was used to construct recombinant plasmids containing four different promoters, i. e., the cytoplasmic actin4 promoter (A4), α-tubulin promoter (α-tub)from silkworm, the Bombyx mori nuclear polyhedrosis virus immediate early protein promoter (IE) and the fibroin heavy chain gene promoter (Fib), respectively. These recombinant plasmids, i. e., pDsRed-A4,pDsRed-α-tub,pDsRed-Fib and pDsRed-IE, had been constructed successfully by restriction enzyme digestion and PCR analysis, and then were transfected into B. mori cell lines (Bm-e-HNU5) by lipid-mediated method to observe the ability of the four promoters to drive RFP reporter gene transient expression in cells. Transfection and transcription experiments indicated that except pDsRed-A4, the other three kinds of recombinant plasmids all transfected Bm-e-HNU5 obviously. The promoters of α-tub, IE and Fib enhanced the transient expression activity of RFP reporter gene in the Bm-e-HNU5, and their activity strengthened sequentially.国家自然科学基金项目(39870410

InFluence of Laser Irradiation on Serum Albumen of Castor Silkworm

我们采用不同波长的激光照射蓖麻蚕蚕卵，使它产生半致死效应，经孵化后的幼虫在五龄期取血分析它的血清蛋白谱，证实了激光照射对蓖麻蚕的遗传性状具有诱变作用，其机理涉及到激光照射与基因表达及调控的关系。Eggs of castor silkworm are irradiated with diFFerent wavelengths of laser and they are made to produce the semilethal eFFect.AFter incubation, taking some blood From larva at the age of Five, the spectra of its serum albumen is analysed.It has been conFirmed that laser irradiation has a mutagenesis on heredity of castor silkworm.the mechanism involved with the relation of laser irradiation with gene expression, as adjuustion and control.福建省自然科学基