地锦和五叶地锦种间杂交不亲和性初步研究

将地锦和五叶地锦杂交,未能获得杂交种。对人工辅助自交和杂交后,花粉萌发和花粉管生长情况进行了荧光显微观测。观察发现：人工辅助自交后,花粉粒可在各自柱头表面正常萌发,花粉管可伸入花柱内。将地锦和五叶地锦杂交时,多数花粉粒不能在柱头表面萌发,或者花粉管呈卷曲、先端膨大、破裂等异常现象。授粉后2～48h的花柱中,未观察到花粉管。由此推断,地锦和五叶地锦之间的不亲和发生在柱头表面。     

Sex Determination of Bovine Embryo Biopsies Using in situ Hybridization*

A male specific bovine DNA fragment was used as probe to determine the sex of bovine interphase cells by in situ hybridization. This method also proved useful for determining the sex of bovine embryos.     

Production of hybrid seed in a seed orchard of two larch species,Larix sibirica and Larix decidua

The larch seed orchard studied is intended to produce interspecific hybrid seed between Larix sibirica and Larix decidua. Only the L. sibirica clone (half of the grafts) in the orchard was intended to act as the seed parent, and the six L. decidua clones are pollinators. The proportion of hybrid seed produced was determined using Mdh3 locus as a genetic marker. The mean percentage of hybrid seed produced by the grafts of L. decidua was 93, but that of the hybrid seed produced by L. sibirica was only 18. The results can be explained by differences in flowering abundance. The grafts of L. sibirica produced several times more pollen than those of L. decidua. Contrary to the original plans, L. decidua should be used as the seed parent and L. sibirica only as the pollen parent in the studied seed orchard.     

毛叶秋海棠的杂交遗传特性

 采用毛叶秋海棠与云南野生秋海棠, 以及国外毛叶秋海棠的园艺品种进行正反交试验, 观测各杂交F1 代群体叶片斑纹、茎的类型、植株抗寒能力等主要性状的表现及其遗传特性; 通过F1 代自交, 对F2 代群体叶片斑纹的观测统计, 研究银绿斑性状的分离规律。试验结果表明: 毛叶秋海棠银绿斑与无斑是一对相对性状, 银绿斑对无斑是显性, 无斑对银绿斑是隐性, 其遗传杂交符合孟德尔的分离规律; B . rex ×B . longialata F1 代植株银绿斑性状的持续表现受光照强度的影响较大, 生长2 年后需要增加约2000 lx 的光照才能保持原有的银绿斑色彩; 毛叶秋海棠根状茎性状的遗传特性极为稳定, 遗传保持率90 %; 杂交F1 代在抗寒性方面表现了明显的杂种优势。     

鸡白痢沙门氏菌与鸡伤寒沙门氏菌基因组消减文库的构建

为了解鸡白痢沙门氏菌与鸡伤寒沙门氏菌的基因组差别,筛选鸡白痢沙门氏菌特有的序列,利用SSH对鸡白痢沙门氏菌C79-13（实验方）与鸡伤寒沙门氏菌Sg9（驱动方）基因组进行了比较。选择四碱基内切酶Rsa I将C79-13与Sg9基因组DNA酶切,酶切的C79-13基因组DNA分成两份,分别与接头1和接头2R连接,然后进行两轮杂交和两轮PCR ,得到消减混合物, 将其与PCR?2.1克隆载体连接,转化感受态E.coli DH5α建立消减文库。结果共获得400个阳性克隆;筛选240个克隆制备质粒进行PCR检测,均扩增出100～2000 bp大小的片段。差异DNA消减文库的构建为进一步筛选、克隆鸡白痢沙门氏菌特异的未知新基因、建立分子鉴别新体系奠定了基础。     

Induction of interspecific allo-tetraploids of Megalobrama amblycephala♀×Megalobrama terminalis♂ by heat shock

Interspecific allo-tetraploidy was induced by a combination of hybridization of Megalobrama amblycephala ♀× Megalobrama terminalis ♂ and thermal shock. A thermal shock at 40 °C for 2 min applied to eggs 33 or 26 min (τ₀: 1.40) after fertilization resulted in a significantly ( P <0.01) higher yield of allo-tetraploids than hybrid crosses without heat shock. The yield of allo-tetraploids in embryos from mid-gastrula to hatching stages was 15.7–24.4% in different thermal shock treatment groups. The allo-tetraploidy rate in 330-day-old juveniles was 9.3% and 6.7% in different year treatment groups. After 2 years of rearing, allo-tetraploidy males attained sexual maturity with milk-like milt at the age of 2+, while allo-tetraploidy females at the age of 2+, and even at 3+, did not show any signs of sexual maturation exhibiting obviously delayed maturity. Only a few allo-tetraploidy females (three individuals) showed maturation characters at the age of 4+ and these females were induced to spawn.     

Identification of differentially expressed genes associated with bud dormancy release in tree peony （Paeonia suffruticosa） by suppression subtractive hybridization

A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies.In order to identify genes that are highly expressed in buds released from dormancy,588 clones were examined by differential screening.Of these,185 clones were selected to be sequenced.A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database.Semi-quantitative RT-PCR was used to confirm further the expression...     

抑制消减杂交（SSH）及其在鱼类基因克隆中的应用

和其它脊椎动物一样,在鱼类的生命进程中,某种特定类型的细胞中只有比例约为10%～15%的基因表达.由于这些基因的特异性决定了从受精卵到机体死亡的整个生命过程,所以这些基因的时空表达是受到严格调控的.在不同发育阶段,不同生理状态和不同类型的细胞中,表达的基因各不相同.因此,比较两种不同类型细胞中基因表达的差异,可以探究表型的遗传原因,了解生命过程的基本信息,从而找到与发育、疾病等相关的基因.     

Male heterogametic sex determination in Rana dybowskii based on sex-linked molecular markers

Identifying the mechanism for sex determination in amphibians is challenging. Very little is known about sex determination mechanisms of Rana dybowskii, a species of importance to evolutionary and conservation biology. We screened for sex-linked molecular markers in R. dybowskii in China using target region amplification polymorphism with 2 fixed primers against the sequences of Dmrt1. We found 2 male-linked molecular markers in R. dybowskii, which were 222 bp and 261 bp long. The detection rates of 222 bp marker in males form Xinglong, Huadian, and Dandong were 93.79%, 69.64%, and 13.64%, respectively, while the rate in females from Huadian was 27.50%. Besides, the detection rates of 261 bp marker in the above 3 regions were only observed in males at the rate of 93.79%, 87.50%, and 32.73%, respectively. The inheritance patterns of sex-linked molecular markers showed that the 2 sex-linked molecular markers were heterozygous. Compared to the XY-male parent, progeny from XX-pseudo-male parent possessed lower sex reversal ratio at the same rearing temperature, and the proportion of female froglets from an XX-pseudo-male parent was more than 95% at low rearing temperature (15°C). Our findings suggest that R. dybowskii displays male heterogamety, and the 2 sex-linked molecular markers may have a guiding significance for the protection and utilization of R. dybowskii.