中国部分板栗品种的SSR标记

从板栗(Castanea mollissima)燕红中分离到25个SSR标记。为了鉴定这些SSR位点,在重复序列的两侧侧翼序列设计引物,并通过化学荧光检测法对6个板栗样品进行检测,共检测到20个多态性位点,每个位点的等位基因数为2~6个。挑选12个位点,通过半自动系统ABI PRISM377对中国北方24个板栗品种进行分析,这12个标记显示了高达75个等位基因的遗传多态性,每个位点的等位基因为4~10个,平均为6.3个,预期的平均杂合性为0.743(介于0.680~0.845),观察值为0.829(介于0.730~0.930)。无效等位基因估算频率显示为3个位点的正向价值,除了一个位点外,这些价值都很低,鉴定几率为7.01×10-11,亲缘关系鉴定几率非常高,为0.999,足够用于花粉流的研究。     

抗生素FR-008基因簇上游区域的DNA序列分析

链霉菌FR-008产生一种七烯大环内酯类抗生素FR-008，对真菌具有很高的抗菌活性，并对蚊子幼虫有高毒性（袁德军和周启，1990）。负责合成FR-008的基因簇被定位（Hu et al．，1994），有21个基因（共138kb，GenBank accession number AY310323）被确认参与了抗生素FR-008的生物合成以及调节和外运（Chen et al．，2003）。然而，在FR-008基因簇最左端fscO基因上游的基因是否与FR-008抗生素生物合成相关？为此，本研究对FR-008基因簇上游区域进行了序列测定和分析。     

柑橘体细胞杂种及其核质遗传研究进展

原生质体融合能够克服柑橘多胚、雌和／或雄性败育等生殖障碍，为柑橘(Citrus spp．)品种改良提供新的途径。柑橘原生质体融合主要采用“叶肉原生质体 胚性愈伤组织原生质体”和“胚性愈伤组织原生质体 胚性愈伤组织原生质体”两种模式，融合方式主要有对称和非对称融合。通过原生质体融合已获得200余个组合的体细胞杂种，杂种倍性多样，绝大多数为异源四倍体，其次为叶肉亲本型二倍体，此外还获得了一些其它倍性的杂种植株。体细胞杂种在柑橘品种改良上的应用主要是砧木改良和用作倍性杂交培育无核三倍体的亲本。柑橘体细胞杂种的鉴定方法有细胞学、形态学和遗传标记。柑橘体细胞杂种核质遗传研究表明，四倍体体细胞杂种核主要为双亲之和，二倍体体细胞杂种核来自叶肉亲本，叶绿体DNA表现出单向或单亲分离，在个别组合中有双亲叶绿体DNA共存现象；线粒体DNA主要来自悬浮系亲本，仅在少数组合中观察到重组或丢失现象。     

茴香脑衍生物的合成工艺研究

以茴香脑为原料经H2O:氧化合成茴香脑环氧化物2-(4-甲氧基苯基)-3-甲基环氧乙烷,产率91%,并研究了溶剂配比、物料配比、催化剂用量、反应温度、反应时间等因素对产物产率的影响.实验结果表明:乙腈与甲醇体积比为1:5、茴香脑与H2O2的物质的量之比为1:2.5、原料与无水碳酸钠的物质的量之比为1:0.8、反应时间为...     

Rapid detection of self-biting disease of mink by specific sequence-characterized amplified regions

Self-biting disease occurred in most farmed fur animals in the world. The mechanism and rapid detection method of this disease has not been reported. We applied bulked sergeant analysis (BSA) in combination with RAPD method to analyze a molecular genetic marker linked with self-biting trait in mink group. The molecular marker was converted into sequence-characterized amplified regions (SCAR) marker for rapid detection of this disease. A single RAPD marker A8 amplified a specific band of 263bp in self-biting minks, which was designated as SRA8-250, and non-specific band of 315bp in both self-biting and healthy minks. The sequences of the bands exhibited 75% and 88% similarity to Canis familiarizes major histocompatibility complex (MHC) class II region and Macaca mulatta MHC class I region, respectively. A SCAR marker SCAR-A8 was designed for the specific fragment SRA8-250 and validated in 30 self-biting minks and 30 healthy minks. Positive amplification of SCAR-A8 was detected in 24 self-biting minks and 12 healthy minks. χ2 test showed significant difference (p<0.01) in the detection rate between the two groups. This indicated that SRA8-250 can be used as a positive marker to detect self-biting disease in minks. Furthermore, the finding that self-biting disease links with MHC genes has significant implications for the mechanism of the disease.     

Polymorphism and heredity of cpDNA and mtDNA in the Section Leuce of <Emphasis Type="Italic">Populus</Emphasis>

The chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) of 16 Populus species (Section Leuce) and their F1 generation were detected using PCR-RFLP technique. The results show that cpDNA in the F1 generation of 22 hybrid combinations was inherited maternally, which supported the conclusions of the study of plasmid cytology. The mtDNA fragments amplified by PCR were consistent with the restriction maps in all hybrid combinations and no polymorphism was detected, indicating that the Section Leuce is highly conserved in mitochondrial gene sequences. These results provided direct evidence of maternal chloroplast inheritance in Populus tomentosa, P. bolleana, P. davidiana, P. adenopoda, P. tomentosa × P. bolleana, P. alba × P. glandulosa and P. alba × P. tomentosa.     

大型真菌DNA提取方法的改进

大型真菌中的一些类群的子实体由于含胶质和多糖较多,对提取高质量的DNA及随后的分子生物学实验有很大影响.针对这一现象,对过去大型真菌的DNA提取方法进行了改进,提出一种实用、高效的DNA提取方法.通过实验证明,此方法去除样品中胶质和多糖的效果明显,可有效获得高质量的DNA,满足随后进行的分子生物学研究需要.     

泰国产方斑东风螺养殖群体遗传多样性的RAPD分析

用随机扩增多态性DNA(RAPD)技术对泰国产方斑东风螺养殖群体的遗传多样性进行检测,从100个随机引物中筛选出21个引物对方斑东风螺的DNA进行扩增,结果表明:21个引物共检测到222条清晰且重复性好的条带,每个引物可扩增出4~16条带,分子量在200~2200bp之间,其中多态位点为156个,占70.27%;群体的Shannon多样性指数为0.2818,Nei基因多样性指数为0.2491;个体间最大遗传距离为0.291,最小遗传距离为0.066。通过与其他贝类遗传多样性的研究结果比较,可初步判断泰国产方斑东风螺养殖群体的遗传多样性比较丰富。     

Impact of soil fumigation practices on soil nematodes and microbial biomass

This study was designed to understand the impact of methyl bromide (MB) (CHaBr) and its alternatives on both free-living and root-knot nematodes in the soil. A randomized complete block experiment with six treatments and 4 replicates (each replicate in a separate greenhouse) was established in Qingzhou, Shandong Province, China. In addition to MB and untreated control (CK) treatments there were four alternative soil fumigation practices including MB virtually impermeable films (VIF), metam sodium (MS), MS VIF and soil solarization combined with selected biological control agents (SS BCA). Two tomato (Lycopersicum esculentum Mill.) cultivars, cv. Maofen-802 from the Xian Institute of Vegetable Science, China, and cv. AF179 Brillante from the Israeli Hazera Quality Seeds, were selected as test crops. The results indicated that Rhabditidae was the most dominant population with percentage abundance as high as 85% of the total number of identified free-living nematodes, followed by that of Cephalobidae. Methyl bromide and its alternatives except for the non-chemical SS BCA treatment controlled the target pest, root-knot nematodes. Also, the impact of the three chemical alternatives on free-living nematode number and functional group abundance was similar to the impact associated with a typical methyl bromide application. Chemical fumigation practices, especially that with MB, significantly reduced the number of nematodes in the soil and simultaneously significantly reduced the number of nematode genera thereby reducing nematode diversity. All the four soil chemical fumigation activities decreased soil microbial biomass and had an obvious initial impact on microorganism biomass. Furthermore, both plant-parasitic and fungivore nematodes were positively correlated with soil microbial biomass.     

连续热裂解法大规模提取质粒DNA

选择质粒保有率高及最适合发酵的TOP10菌株,在传统的煮沸裂解法提取质粒的基础上进行了提取方法的改进,使用恒流泵和热交换螺旋管,使菌体在流动过程中完成热裂解,释放质粒DNA。改进后的方法裂解温度可降到70",提取工艺流程简单,缩短了提取时间,提高了质粒DNA的提取效率。利用本方法提取的质粒大小正确、纯度合格,每升发酵液得率平均为100mg质粒DNA。