四种鱼类外周血红细胞细胞周期及DNA含量

通过流式细胞仪测量了兴国红鲤（Ｃｙｐｒｉｎｕｓ ｃａｒｐｉｏ Ｖａｒ． Ｓｉｎｇｕｏｎｅｎｓｉｓ）、奥利亚罗非鱼（Ｓａｒｏｔｈｅｒｏｄｏｎ ａｕｒｅａ）、 鳙（Ａｒｉｓｔｉｃｈｔｈｙｓ ｎｏｂｉｌｉｓ Ｒ．）和团头鲂（Ｍｅｇａｌｏｂｒａｍａ ａｍｂｌｙｃｅｐｈａｌａ Ｙｉｎ．）红细胞的ＤＮＡ含量,四种鱼与鸡红细胞ＤＮＡ含量的比值分别为１·６５、０．９６、０．９１、１·１５,以鸡红细胞ＤＮＡ含量２．３ｐｇ／Ｎ计算,四种鱼二倍体体细胞的 ＤＮＡ含量分别为３．８０ ｐｇ／Ｎ, ２． ２２ｐｇ／Ｎ、２． ０８ｐｇ／Ｎ、 ２． ６６ｐｇ／Ｎ。另外,四种鱼外周血红细胞分别有２６％、 ２６％、 ２３％、 ２４％的细胞处于 ＤＮＡ合成期（Ｓ）,ＤＮＡ合成后期（Ｇ２）和分裂期（Ｍ）,这表明这四种鱼类的外周血红细胞并非失去分裂能力的特化细胞群,它们表现了较强的细胞周期现象。这有可能是这几种硬骨鱼类外周血仍具有造血功能的缘故。     

几种鲟鱼基因组大小、倍体的特性及鲟形目细胞进化的探讨

采用Ｆｅｕｌｇｅｎ－显微分光光度计方法,以鸡红细胞为标准ＤＮＡ（３．２２ｐｇ／２Ｃ）测定了长江白鲟、达氏鲟、中华鲟、史氏鲟和北美匙吻鲟的体细胞基因组大小（ＤＮＡ含量）。结果表明,上述五种鲟鱼ＤＮＡ含量分别为４．１１、８．２６、９．０７、６．０７和３．９６ｐｇ。长江白鲟和北美匙吻鲟均属于四倍体鱼类,分布在长江水系的中华鲟和达氏鲟两种鲟属鱼类为八倍体类型。史氏鲟初步判断为八倍体,据分析可能存在四倍体的     

人r型基因工程干扰素中残余DNA的检测

采用地高辛标记探针,以核酸杂交法检测人ｒ型基因工程干扰素（ＩＦＮ－ｒ）制品中残余ＤＮＡ含量,结果表明,自制的二组ＤＮＡ标准品相应量间显示的色斑相差悬殊,提示高蛋白含量对痕量残余ＤＮＡ的检测干扰较大,添加ＩＦＮ－ｒ的ＤＮＡ标准品比纯ＤＮＡ标准品更合理,以此测得各样品均符合ＷＨＯ要求（〈１００ｐｇ／剂量）,方法敏感性为４ｐｇ。     

地高辛标记核酸探针斑点杂交法检测轮状病毒疫苗中残余MA－104细胞DNA含量的研究

本文报导用地高辛标记核酸探针和分子斑点杂交技术检测轮状病毒基因重配株Ｌ－３株活疫苗中残余ＭＡ－１０４细胞ＤＮＡ含量的方法。提取和纯化ＭＡ－１０４细胞ＤＮＡ,将其ＡｌｕＩ酶切片段用随机引物法引导ＤＮＡ标记为探针。待检样品抽提核酸后点膜进行斑点杂交。此法灵敏度高,可检出０．１４ｐｇ的ＤＮＡ,特异性强,与非同源性ＤＮＡ无杂交。用此法检测轮状病毒基因重配株Ｌ－３株制备的疫苗,ＭＡ－１０４细胞残余ＤＮＡ含量低于１４ｐｇ低于ＷＨＯ限量标准,结果表明此重配株用于研制疫苗是安全的。     

检测CHO基因工程乙肝疫苗残余DNA方法的建立和应用

对样品的处理方法进行了探索,最后选用一个疫苗剂量的蛋白酶Ｋ同时消化样品和标准ＤＮＡ,并将消化处理后的样品和标准ＤＮＡ用地高辛标记的探针进行检测,灵敏度达１０ｐｇ。共检测２１批检品,除一批ＤＮＡ残余量超过１００ｐｇ／ｄｏｓｅ以外,其它２０批样品的ＤＮＡ残余量均低于１００ｐｇ／ｄｏｓｅ。     

天青Ⅰ显示细胞核DNA组织化学新技术及其应用

天青Ⅰ显示细胞核ＤＮＡ组织化学新技术及其应用田玉旺邢惠清邓永江黄晓南（北京军区总医院病理科北京１００７００）细胞核ＤＮＡ含量的测定是直接反映肿瘤增殖能力的重要生物学指标。目前肿瘤细胞核ＤＮＡ图像定量分析已成为一门新兴的检测技术并广泛应用于临床,常采用...     

稻胚发育过程中细胞核内DNA增殖的特点

应用显微分光光度技术测定的稻胚发育过程中核ＤＮＡ含量的结构表明,球形胚中细胞核ＤＮＡ含量较高,多倍性明显,Ｃ值较集中在６－１２Ｃ,个别的最高达２３８或４３９Ｃ,它们可能为胚柄细胞;ＤＮＡ含量差异明显可能与它们的分化潜能有关。分化初期,中期及成熟胚细胞核ＤＮＡ含量一般不及球形胚高,不同倍性的频率分布亦较球形胚分散,但多倍性仍明显;     

普通小麦三个基因组之间的遗传关系及原位杂交分析

以普通小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍＬ．）的３个可能的二倍体供体种（乌拉尔图小麦（Ｔ．ｕｒａｒｔｕＴｈｕｍ．）拟斯卑尔脱山羊草（ＡｅｇｉｌｏｐｓｓｐｅｌｔｏｉｄｅｓＴａｕｓｃｈ）和粗山羊草（Ａｅ．ｔａｕｓｃｈｉｉＣｏｓｓ．）的基因组ＤＮＡ为研究对象,通过它们之间的相互杂交,比较杂交强度以及泳道中带纹的不同,并结合部分ＤＮＡ重复序列在基因组间含量差异的数据,得出结论：Ａ＾ｕ和Ｄ基因组的关系     

用地高辛标记探针检测基因工程人β干扰素制品中的外源DNA

基因工程人β干扰素工程菌经发酵培养、纯化后获得纯的制品,采用斑点杂交技术,用非放射性地高辛标记超声裂解的全工程菌ＤＮＡ作为探针,检测基因工程人β干扰素纯品,结果显示：基因工程人β干扰素纯品中的外源ＤＮＡ含量小于１００ｐｇ／剂。     

形态测量对肝细胞肝癌分级的研究

应用自行设计组装的ＭＩＰＳ－Ｉ型多功能图像分析仪对６０例不同级的肝细胞肝癌细胞核形态学以及ＤＮＡ含量的９个参数进行了测定。经多元判别分析建立判别方程,发现９个参数都有判别能力。对判别方程贡献最大的四个参数依次为核面积、核平均光密度＞５Ｃ细胞百分数和核周长。回代符合率为９５％。     

Nuclear DNA content of Vitis vinifera cultivars and ploidy level analyses of somatic embryo-derived plants obtained from anther culture

Flow cytometry was employed to determine the ploidy level of Vitis vinifera L. somatic embryo-derived plants obtained from anther culture. Only one among the 41 analysed plants (2.4%) presented somaclonal variation (tetraploidy); the other plants were diploid. No significant differences (P≤0.05) were detected between diploid and parental field plants. No haploid or aneuploid plants were observed. The nuclear DNA content of nine V. vinifera cultivars was also estimated using flow cytometry. A non-significant variation was found among the cultivars, with DNA content ranging from 1.17 pg/2C (cv. ‘Tinta Barroca’ and ‘Viosinho’) to 1.26 pg/2C (cv. ‘Cabernet Sauvignon’). These results and previous studies on other Vitis species suggest that Vitis genome is stable with regard to nuclear DNA content.     

Rapid detection of aneuploidy in <Emphasis Type="Italic">Musa</Emphasis> using flow cytometry

We report a procedure for the rapid and convenient detection of aneuploidy in triploid Musa using DNA flow cytometry. From a population of plants derived from gamma-irradiated shoot tips, plants were selected based on aberrant morphology and their chromosome numbers were counted. Aneuploids plants with chromosome numbers 2n=31 or 32 were found as well as the expected triploid plants (2n=3x=33). At the same time, the nuclear DNA content of all plants was measured using flow cytometry. The flow cytometric assay involved the use of nuclei isolated from chicken red blood cells (CRBC), which served as an internal reference standard. The relative DNA content of individual plants was expressed as a ratio of DNA content of CRBC and Musa (DNA index). In order to estimate the chromosome number using flow cytometry, the relative DNA content of plants with unknown ploidy was expressed as a percentage of the DNA content of triploid plants. The classification based on flow cytometry fully agreed with the results obtained by chromosome counting. The results indicated that flow cytometry is a convenient and rapid method for the detection of aneuploidy in Musa.     

Variation in nuclear DNA content in Malus species and cultivated apples.

The nuclear DNA content for a group of 40 Malus species and hybrids has been estimated using flow cytometry. Estimates of nuclear DNA content for this germplasm collection range from 1.45 pg for Malus fusca (diploid) to 2.57 pg for Malus ioensis (triploid). Among diploids, the nuclear (2C) DNA ranges from 1.45 pg for M. fusca to 1.68 pg for Malus transitoria. Among triploids, the nuclear (3C) DNA content ranges from 2.37 pg / 3C for Malus sikkimensis to 2.57 pg / 3C for M. ioensis. Given the complexity of the apple genome and its suggested allopolyploid origin, the results obtained in this study confirm earlier reports that polyploids can easily withstand the loss of a certain amount of DNA, and that there is a slight tendency towards diminished haploid nuclear DNA content with increased polyploidy.     

Presence of triploids among oak species

BACKGROUND AND AIMS: Polyploids, organisms that have multiple sets of chromosomes, are common in certain plant and animal taxa. However, there are only a few reports of intraspecific ploidy variation within the genus Quercus. The aim of the study was to investigate the suspected ploidy level of two oaks that have unusual microsatellite banding patterns. METHODS: Polyploidy was investigated by using microsatellite analysis, stomata length measurements and nuclear DNA content estimation by flow cytometry. KEY RESULTS: Each putative triploid tree has patterns of microsatellite variation unexpected for diploid genomes, with up to three alleles at some loci, significantly longer stomata and 1.5 times more DNA per nucleus compared with diploids. CONCLUSIONS: To our knowledge, this report contains the first evidence for triploidy in Q. petraea and confirmation of this phenomenon in Q. robur. Regardless of the positive or negative aspects of the presence of triploid oaks in forest stands, it is of value to be able to screen for them. This study demonstrates that nuclear microsatellites and estimation of DNA content by flow cytometry can readily be used for this purpose.     

Nuclear DNA content and base composition in 28 taxa of Musa.

The nuclear DNA content of 28 taxa of Musa was assessed by flow cytometry, using line PxPC6 of Petunia hybrida as an internal standard. The 2C DNA value of Musa balbisiana (BB genome) was 1.16 pg, whereas Musa acuminata (AA genome) had an average 2C DNA value of 1.27 pg, with a difference of 11% between its subspecies. The two haploid (IC) genomes, A and B, comprising most of the edible bananas, are therefore of similar size, 0.63 pg (610 million bp) and 0.58 pg (560 million bp), respectively. The genome of diploid Musa is thus threefold that of Arabidopsis thaliana. The genome sizes in a set of triploid Musa cultivars or clones were quite different, with 2C DNA values ranging from 1.61 to 2.23 pg. Likewise, the genome sizes of tetraploid cultivars ranged from 1.94 to 2.37 pg (2C). Apparently, tetraploids (for instance, accession I.C.2) can have a genome size that falls within the range of triploid genome sizes, and vice versa (as in the case of accession Simili Radjah). The 2C values estimated for organs such as leaf, leaf sheath, rhizome, and flower were consistent, whereas root material gave atypical results, owing to browning. The genomic base composition of these Musa taxa had a median value of 40.8% GC (SD = 0.43%).     

Molecular cytogenetic study on the ploidy status in Acipenser mikadoi

The ploidy status of Acipenser mikadoi was examined using nuclear DNA contents, karyotypes and fluorescence in situ hybridization (FISH) with 5.8S + 28S rDNA as a probe. In flow‐cytometrically sorted specimens with 8.2–9.1 pg DNA content per somatic cell, i.e. genetic diploid, the best informative metaphase with 268 chromosomes had 80 biarmed meta‐ or submetacentric (M or SM) chromosomes, 48 monoarmed telocentric (T) chromosomes and 140 microchromosomes. In genetic triploid specimens with 12.6–13.0 pg DNA content, the best informative metaphase with 402 chromosomes showed 120 biarmed M or SM, 72 monoarmed T chromosomes and 210 microchromosomes. The rDNA FISH detected a maximum 18 and 27 signals in the diploid and triploid A. miakdoi, respectively. The obtained findings thus corroborated a clear parallel between nuclear DNA contents and karyological or FISH profiles in the genetic diploid and triploid specimens, suggesting 1.5 times chromosome complements of diploid counterparts or three sets of homologues in the triploid sturgeons. Moreover, the estimated genome size and the observed molecular cytogenetic features in the diploid A. mikadoi strongly suggest that this species is a member of a functional tetraploid group recently proposed in the literature.     

应用流式细胞术测定17种中国野生蔷薇核DNA含量

以17种中国野生蔷薇为试材，采用改良的LB 01裂解液，以4种不同的标准植物——大豆（Glycine max Merr.‘Polanka’）、绿豆（Vigna radiata（L.） Wilczek）、番茄（Lycopersicon esculentum Miller）和玉米（Zea mays L.）为外标，以二倍体材料丽江蔷薇（Rosa×lichiangensis Yü et Ku）为内部参照，利用流式细胞术对其核DNA含量及染色体倍性进行检测，并采用常规染色体压片法验证倍性准确性。本研究首次检测了3个二倍体种——商城蔷薇（Rosa shangchengensis T.C.Ku）、广东蔷薇（Rosa kwangtungensis Yü et Tsai）和无刺刺梨（Rosa roxburghii f.inermis S.D.Shi），1个三倍体种——伞房蔷薇（Rosa corymbulosa Rolfe）和1个四倍体种——弯刺蔷薇（Rosa beggeriana Schrenk）的核DNA含量及基因组大小。结果表明，流式细胞术检测结果与常规染色体压片法结果一致，可对中国野生蔷薇的倍性研究进行补充。本研究结果可丰富中国蔷薇属植物的细胞遗传学背景资料并为繁育新品种提供理论依据。     

甜叶菊同源四倍体离体诱导及鉴定

用不同浓度秋水仙素溶液处理甜叶菊不定芽,诱导同源四倍体,并进行解剖学、染色体鉴定和流式细胞仪鉴定倍性。结果表明:(1)用0.20%的秋水仙素溶液浸泡甜叶菊不定芽12h,同源四倍体诱导率最高,可达32.14%。(2)同源四倍体植株与二倍体(对照)相比,其气孔、叶片等均表现巨大性,且叶片变厚、叶色浓绿、叶片皱缩。(3)对照植株染色体2n=2x=22,四倍体植株染色体2n=4x=44;流式细胞仪倍性鉴定结果显示,对照DNA相对含量为100,四倍体DNA相对含量为200。(4)该研究共鉴定出48株甜叶菊同源四倍体植株,为进行倍性植株的诱导奠定了技术基础,为进一步开展甜叶菊同源四倍体新品种的选育提供了实验材料。     

Ultramicrochemical determination of nucleic acids in individual cells using the Zeiss UMSP-I microspectrophotometer. Application to isolated rat hepatocytes of different ploidy classes

Synopsis Edström's method for the ultramicrochemical determination of RNA and DNA in individual cells was modified for the measurement of extinction in u.v. light with the aid of the Zeiss scanning microspectrophotometer UMSP-I. With this new procedure, nucleic acids down to about 3 pg RNA or about 4 pg DNA can be measured with a very high accuracy.The method was applied to enzymatically isolated rat liver parenchymal cells. A mean DNA content of 6.52 pg was found for diploid cells. The DNA content of mononuclear cells of different ploidy levels and of binuclear cells showed a close proportionality with the nuclear ploidy and the number of nuclei per cell. The RNA content of mononuclear diploid cells amounted to 33.4 pg, yielding an RNA/DNA ratio of 5.12. The RNA/DNA ratio was similar for binuclear and mononuclear cells of the same ploidy level but decreased considerably with increasing nuclear ploidy.     

Detection of protoplast-derived DNA tetraploid Lisianthus (Eustoma grandiflorum) plants by leaf and flower characteristics and by flow cytometry

Plants of lisianthus (Eustoma grandiflorum (Griesbach)Schinners=Lisianthus russellianus Hook.) were regenerated from protoplasts and grown in pots until flowering. Vegetative and floral characteristics were measured and compared with parent plants. Larger leaves and petals and longer guard cells, sepals and filaments were recorded from protoplast-derived plants suggestive of polyploidy. The nuclear DNA contents of protoplast-derived and parental plants were determined by flow cytometry. Protoplast-derived plants were confirmed as DNA tetraploid by flow cytometry with a DNA index of 1.95. Their nuclear DNA content was measured as 6.33±0.04 pg DNA per 2C nucleus compared with 3.26±0.10 pg DNA per 2C nucleus from parental plants. Polyploidisation induced during protoplast regeneration offers an alternative to that of colchicine treatment.