流式细胞术在水生生物DNA含量和倍性分析中的应用

利用流式细胞仪对鲤鱼、银鲫、黄颡、牙鲆等淡水和海水鱼类进行了DNA含量测定或倍性分析。通过几百个样品的测定和分析,认为流式细胞仪在DNA相对含量测定和倍性分析上结果比较稳定一致,可以用于大量样品的测定分析。同时对待测样品的保存、处理及上样量等进行了一系列比较研究,找出最好的保存方法、最简便的样品处理方法和最少的上样量。流式细胞仪在水生生物的遗传育种、种群分化等方面有很重要的应用价值。     

Ecology,physiology and genetics of a phycodnavirus infecting the noxious bloom-forming raphidophyte <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis>

Abstract:   Heterosigma akashiwo virus (HaV) is a large icosahedral virus (∼0.2 μm) harboring a double-stranded DNA (dsDNA) genome (∼294 kbp). The virus is the only member of the genus Raphidovirus in the family Phycodnaviridae. Since its first discovery, a number of ecologic, physiologic and genetic studies about HaV have been conducted; especially, the relationship between H. akashiwo and HaV in nature was studied and viral infection is now regarded as a significant factor influencing the dynamics and termination of H. akashiwo blooms. HaV infection has considerable impacts on H. akashiwo populations in both aspects of fluctuation in biomass (quantity) and changes in clonal composition (quality). Partial sequencing of the HaV genome revealed that a number of genes showed considerable similarity to those of other protist-infecting viruses; still, the phylogenetic position of HaV suggested a number of enigmas in host–virus coevolution. Here are summarized the ecology, physiology and genetics of HaV especially from the viewpoint of the host–virus relationship.     

Direct evidence of multiple paternities in natural population of viviparous Japanese surfperch by allelic markers of microsatellite DNA loci

This study was performed to obtain information on the occurrence of multiple paternities in three species of viviparous Japanese surfperch using allelic markers of microsatellite DNA loci. Direct evidence for multiple fertilizations was established by reconstructing paternal genotypes from the progeny of gravid females. Multiple paternities were ascertained in five of 10 broods of Ditrema temmincki and in three of nine broods of Neoditrema ransonneti, but not in Ditrema viride. The number of patrilines detected in the progeny of D. temmincki and N. ransonneti females were two or three, respectively, as determined by the GERUD v2.0 algorithm for reconstructing parental genotypes from half-sib progeny arrays.     

柑桔体细胞融合再生9个组合的二倍体叶肉亲本类型植株

电场诱导９个柑桔种间及体内体细胞融合,各组合均再生叶肉原生质体亲本类型植株。这些植株经形态学和细胞学检查证明为二倍体（２ｎ＝２ｘ＝１８）。对其中５个组合进行ＲＡＰＤ分析表明,４个组合植株的谱带在所分析的引物上均表现为与叶肉细胞亲本谱带一致,另１个组合,即Ｐａｇｅ柑柚＋粗柠檬,个别植株除含有叶肉亲本的所有谱带外,还扩增出了悬浮系亲本的部分特征带,为杂种类型。探讨了这类植株的产生原因及潜在价值。     

Mitochondrial DNA deletion of the brain tissue of aged rats with learning and memory deficit

AIM and METHODS: The ratio of mitochondrial DNA (mtDNA) deletion was measured to find the relationship between mtDNA deletion and aged learning and memory deficit. The aged rats were divided into two groups, aged learning and memory deficit group and aged learning and memory normal group. The ratio of mtDNA deletion was measured by dilution polymerase chain reaction. RESULTS: There are deleted mtDNA (about 4834 bp) in the cerebral cortex, hippocampus and cerebellum of both young and aged rats. The ratios of deleted mtDNA were similar in the cerebral cortex,hippocampus and cerebellum of young rats (about 0.00018%). The ratio mtDNA of aged learning and memory normal rats had increased by five-fold in the cerebral cortex and hippocampus, or one-fold in the cerebellum over young rats. The ratio of aged learning and memory dificit rats had increased by one-fold in the cerebral cortex or 0.8-fold in the hippocampus or two-fold in the cerebellum over aged learning and memory normal rats.CONCLUSIONS: There was really the increase of mtDNA in aging rat brain. And this increase was double in amount in aged learning and memory deficit rats compared to the normal learning and memory aged rats. It is suggested that the mtDNA deletions in the brain regions associated with learning and memory may be contributed to the cellular and molecular mechanism of learning and memory deicit with aged rats.     

Activation of repair pathways after neuronal DNA damage induced by bupivacaine

AIM To investigate the activation of related repair pathways after bupivacaine-induced neuronal DNA damage by cDNA gene screening. METHODS The bupivacaine-induced SH-SY5Y neuronal damage and DNA damage model was established. The technique of cDNA microplate array was used to screen the 21 important regulatory factors in the DNA damage repair pathway. Post-analysis of these differentially expressed repair genes for the repair pathway enrichment and distribution was performed. The data were analyzed by GraphPad Prism 6 statistical software to compare differences between groups. RESULTS The viability of SH-SY5Y cells treated with bupivacaine at different concentrations （detected by CCK-8 assay） showed that the IC₅₀ value of bupivacaine was 1.5 mmol/L. The comet assay related index （the comet tail） was increased （P<0.05）, the phosphorylation level of γH2AX protein was increased （P<0.05）, indicating that DNA damage in the SH-SY5Y cells was significantly aggravated after bupivacaine treatment. The results of cDNA microplate assay showed that compared withcontrol group, the differentially expressed genes after bupivacaine treatment were DNA-PKcs, PTEN, NTH1, RAD9, CSB, GADD45, XPD, XPC-HR23B and P53. The analysis showed that these repair genes were mainly concentrated in the following 3 repair mechanisms： base excision repair, nucleotide excision repair, and non-homologous reconstitution. CONCLUSION The repair genes differentially expressed after neuronal DNA damage caused by local anesthetics are mainly concentrated in the pathways of non-homologous end-joining, base excision repair and nucleotide excision repair.     

苹果RAPD分析体系的建立

对富士苹果（Ｍａｌｕｓ　ｐｕｍｉｌａ　Ｍｉｌｌ．ｃｖ．Ｆｕｊｉ）ＲＡＰＤ（Ｒａｎｄｏｍ　Ａｍｐｌｉｆｉｅｄ　Ｐｏｌｙｍｏｒｐｈｉｃ　ＤＮＡ）分析体系的优化研究表明,ＲＡＰＤ反应体系中,ＤＮＡ、Ｔａｑ酶、引物和Ｍｇ２＋４种主要成分的最适用量分别为：２０ｎｇ、１．０ Ｕ、０．２ｕｍｏｌ·Ｌ－１和３．０ｕｍｏｌ·Ｌ－１。采用该优化体系,以 ＯＰＪ０３为引物,构建了我国及世界范围内苹果生产中重要的２８个品种的ＲＡＰＤ指纹图谱,分析了其遗传多样性,区分了供试的２８个苹果品种中的１５个,区分率达５３．６％。讨论了ＲＡＰＤ鉴定苹果品种的应用及其主要影响因素。     

大豆对大豆胞囊线虫侵染的应答机制研究

大豆胞囊线虫病(Heterodera glycines,soybean cyst nematode,SCN)是大豆生产上的重要病害,其特点为危害重、分布广、难防治,每年对大豆生产造成极大的损失。种植大豆抗性新品种是防治SCN目前最为有效的措施,研究大豆对SCN侵染的应答机制,是培育大豆持久抗病品种的前提,对加快抗线虫品种选育及SCN的防控具有重要的意义。本文综述了大豆对SCN侵染的组织细胞学应答机制;介绍了大豆在SCN侵染后酶系变化及酚类代谢的生理生化应答机制;从分子水平阐明了SCN侵染后大豆的基因转录变化,差异蛋白及DNA甲基化的应答机制,以期为大豆胞囊线虫病害的进一步研究与防治提供参考。