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

GENETYPE STUDY ON A NEW TRANSLUCENT WHITE EGG MUTANT of SILKWORMS

国家自然科学基

Genetical Research of BT924—A New Line in White Egg Mutants of Silkworm

新的家蚕白色卵突变系BT924是由蓖麻蚕DNA导入家蚕品种苏学5诱导获得。性状特征 :滞育卵当年白色 ,越年后呈浅黄褐色 ;蛾区内及区间卵色略有变异 ,幼虫皮肤低度透明 ,成虫复眼黑色。采用家蚕卵色正常型 (黑卵 )和突变型红色卵 (re)、桃红眼白卵 (pe)、第2白卵 (w -2)、第3白卵 (w-3)及BH863油蚕白卵 (w -3bh)与之杂交 ,进行遗传分析 ,结果表明 ,BT924白卵及其油蚕性状由隐性单基因控制 ,普通遗传 ,基因座与w-3相同 ,即 :10 -19.6。命名为 :whiteeggBT924 ,基因符号 :w -3bt。w -3bt对re表现上位作用。同时发现家蚕基因库保存的桃红眼白卵 (pe)标记基因系pe000同时持有红色卵 (re)基因 ,其卵色基因型为repe/repe。A new line in white egg mutants of silkworm (Bombyx mori)—BT924 was derived from incorporation of eri silkworm (Philosamia cynthia ricini Donovan) DAN into the silkworm race “Su Xue 5”.It has the following characteristic traits:The diapause eggs are white in color in the hiberating year and become fawn after hiberation; Slight intra and inter batch variation in gee color may exist; Larval skin is slightly translucent; And the compound eyes of the imagos are black.In the present research,the new white egg mutant line was crossed with a normal silkworm race(black egg) and with the mutant lines re,pe,w-2,w-3 and w-3bh for genetical analysis.White egg of BT924 together with its translucency was shown to be controlled by a single recessive gene and share the same locus with w-3(10-19.6). It has been named“white egg BT924”with the gene symbol w-3bt. w-3bt is epistatic to re.In addition,the marker gene line pe000 of pe maintained in the laboratory was shown to carry the re gene as well,its genotype being re pe/re pe.国家自然科学基金资助!(项目编号 :39970570

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

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

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

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

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

灭活 CVB1 在离体骨骼肌细胞中基因转移的研究

【目的】研究灭活后的柯萨奇病毒 B1 在原代培养骨骼肌细胞中对报道基因 pCDNA3-LacZ 转移效率的影响。【方 法】①用聚乙二醇沉淀和蔗糖低温超速离心法提纯并用β-丙内酯灭活病毒; ②分别用不同浓度的转铁蛋白聚赖氨酸( TfpL)+ LacZ、灭活 CVB1+LacZ 以及灭活 CVB1+TfpL+LacZ 在肌肉细胞进行基因转移, 以报告基因产物 β-半乳糖苷酶染色阳性的 细胞数作为基因转移效率指标。【结果】灭活 CVB1+TfpL+LacZ 在肌肉细胞中使基因转移效率由 TfpL +LacZ 组的不到 1%提高到 15%～ 20%。【结论】灭活 CVB1 也能象腺病毒一样促进基因转移,CVB1 的嗜肌肉特性为在肌肉组织中研究靶向 明确的新载体提供了实验依据