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Chromosomal distribution patterns of global 5mC and 5hmC on the ZZ/ZW and XX/XY chromosomes in the Japanese wrinkled frog, Rana rugosa, induced by Tet methylcytosine dioxygenase enzymes Tet甲基胞嘧啶双加氧酶诱导的日本皱蛙ZZ/ZW和XX/XY染色体上5mC和5hmC的染色体分布模式
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.15
Musashi Kubiura, I. Miura, M. Tada
Tet methylcytosine dioxygenase enzymes catalyze the conversion of DNA methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), thereby initiating DNA demethylation. In mice, Tets also regulate gene activity by binding the regulatory elements of target genes. 5hmC is associated with euchromatic regions and epigenetic reprogramming events. In this study, we examined the levels of 5mC and 5hmC in two major groups of Japanese wrinkled frogs (Rana rugosa) with morphologically distinct sex chromosome constitutions, ZZ/ZW and XX/ XY. Patterns of 5mC and 5hmC, as determined by immunostaining, were indistinguishable between homologous sex chromosomes in ZZ cells and XX cells. Genome-wide conversion of 5mC to 5hmC, followed by cell cycle-mediated dilution, was induced in PHA-stimulated peripheral blood cells in adult frogs. Tet enzymes may share an evolutionarily conserved function associated with a global epigenetic reprogramming event during dedifferentiation in mammals and frogs.
Tet甲基胞嘧啶双加氧酶催化DNA甲基胞嘧啶(5mC)转化为5-羟甲基胞嘧啶(5hmC),从而启动DNA去甲基化。在小鼠中,Tets也通过结合靶基因的调控元件来调节基因活性。5hmC与常染色质区和表观遗传重编程事件有关。在这项研究中,我们检测了两种主要的日本皱蛙(Rana rugosa)的5mC和5hmC水平,它们具有形态上不同的性染色体构成,ZZ/ZW和XX/ XY。免疫染色法测定的5mC和5hmC在ZZ细胞和XX细胞的同源性染色体间无法区分。在pha刺激的成年蛙外周血细胞中,5mC全基因组转化为5hmC,随后进行细胞周期介导的稀释。Tet酶可能在哺乳动物和青蛙去分化过程中共享一个与全局表观遗传重编程事件相关的进化保守功能。
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引用次数: 0
Chromosome instability of allopolyploid resynthesized Brassica napus 异源多倍体再合成甘蓝型油菜的染色体不稳定性
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.79
N. Ohmido, Kana Ueda, K. Fujii
Allopolyploid resynthesized Brassica napus (AACC, 2n = 38) was produced by cross-hybridization between B. rapa (AA, 2n = 20) and B. oleracea (CC, 2n = 18) for new vegetative crop breeding. Many studies have provided evidences for the phenotype instability and close relationship between A and C genome in the amphidiploid resynthesized B. napus cultivars. In fact, a new B. napus varieties, Hanakkori showed atypical morphological characters and generated many off-type progeny plants. In this study, we investigated the pollen fertility, chromosome number, structure, and behavior linked to defi ne the factors, which produce the unstable phenotypic expressions in the Hanakkori progenies. We define the chromosome number and chromosome organization using fl uorescence in situ hybridization analysis on somatic mitosis and meiosis cells. Off-type plants with lower pollen fertility show the instability of chromosome number and structures with small chromosome fragments. Observation of chromosomes behaviors at meiosis showed that the meiotic division in off-type plants led to appreciably higher abnormalities than in on-type plants. These results demonstrated that minimized abnormal chromosome structure and formation might be essential to stabilize the normal progenies in the B. napus plant breed-
利用rapa (AA, 2n = 20)与甘蓝(CC, 2n = 18)杂交获得异源多倍体重组甘蓝型油菜(AACC, 2n = 38),用于培育新的营养作物。许多研究都证明了雌雄二倍体再合成甘蓝型品种的表型不稳定性和A基因组与C基因组的密切关系。事实上,甘蓝型油菜新品种Hanakkori表现出非典型的形态特征,并产生了许多异型后代植株。本研究从花粉的育性、染色体数目、结构和行为等方面探讨了影响花楸后代不稳定表型表达的因素。我们用荧光原位杂交分析确定体细胞有丝分裂和减数分裂细胞的染色体数目和染色体组织。花粉育性较低的off型植物表现出染色体数目和结构的不稳定性,染色体片段较小。对染色体减数分裂行为的观察表明,离型植物的减数分裂导致的染色体异常明显高于同型植物。这些结果表明,尽量减少异常染色体的结构和形成可能是稳定甘蓝型油菜正常后代的关键
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引用次数: 3
Karyological observations on dwarf individuals of Pilea brevicornuta Hayata (Urticaceae) growing in the riparian habitat in Amami-Oshima of the Ryukyus, Japan 日本琉球天美大岛滨岸生境短叶荨麻(荨麻科)矮生个体的核学观察
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.3
Nure Ferdousee, T. Denda, M. Yokota
Cytotaxonomical studies were conducted on Pilea brevicornuta Hayata in the Ryukyu Islands and Taiwan. Karyomorphology of dwarf and creeping individuals growing in riparian habitat of Amami-Oshima Island, which is morphologically distinct from normal individuals growing in forest floor, is the main focus of this report. All of the studied individuals of P. brevicornuta including dwarf ones had same chromosome number of 2n = 24. The karyotype formula is 2n = 24 = 6m + 13sm + 5st for normal individual and 2n = 24 = 6m + 12sm + 6st for dwarf individual. The chromosomes of P. brevicornuta were all small with sizes varying between 0.9 and 3.0 μm. Considering the basic chromosome number in the genus Pilea, 2n = 24 is regarded as diploid based on x = 12. Despite the high variability in morphology, P. brevicornuta had remarkably uniform karyomorphology.
摘要对琉球群岛和台湾的短花椒(Pilea brevicornuta Hayata)进行了细胞分类研究。本文重点研究了鸭美大岛滨水生境中生长的矮生和匍匐个体与生长在森林地面上的正常个体在形态上的差异。所有研究个体,包括矮生个体,染色体数目均相同,均为2n = 24。正常个体核型公式为2n = 24 = 6m + 13sm + 5st,矮秆个体核型公式为2n = 24 = 6m + 12sm + 6st。短花椒的染色体都很小,大小在0.9 ~ 3.0 μm之间。考虑到Pilea属的基本染色体数目,根据x = 12, 2n = 24被认为是二倍体。尽管在形态上有很大的变异,但短花椒的核形态却非常一致。
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引用次数: 0
The role of phosphorylation of histone H3 at serine 10 in chromatin condensation in vitro 组蛋白H3丝氨酸10位点磷酸化在体外染色质凝聚中的作用
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.9
I. Equilibrina, Elena Krayukhina, K. Inoue, Y. Ishikawa, A. Kawamoto, Takayuki Kato, I. Suetake, S. Tajima, H. Takata, S. Uchiyama, K. Fukui
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引用次数: 4
Unusual sex-ratios and developmental mortality in the rice frog Fejervarya kawamurai 稻蛙不寻常的性别比例和发育死亡率
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.53
I. Miura, H. Ohtani, Takeshi Fujitani
In the Japanese rice frog, Fejervarya kawamurai, we identifi ed two unusual features of the sex ratio using artificial crosses at laboratory. Firstly, inbreeding such as sibling-sibling crosses, backcrosses, and diploid gynogenesis resulted in a male-biased sex ratio with a high rate of developmental mortality. Secondly, outbreeding reduced the mortality, and particularly outbreeding of females with males from a geographically separate population restored the sex ratio. Established mechanisms of genetic and environmental sex determination in vertebrates do not easily explain these results. Ecologically, this mechanism favors expanding populations that invade new habitat, because frogs must move continuously between populations to produce enough daughters and reduce embryonic mortality.
在日本稻蛙(Fejervarya kawamurai)中,我们通过实验室人工杂交发现了性别比例的两个不寻常特征。首先,近亲繁殖,如兄弟姐妹杂交、回交和二倍体雌核发生,导致雄性偏倚的性别比例和高的发育死亡率。其次,近亲繁殖降低了死亡率,特别是雌性与来自地理上不同种群的雄性的近亲繁殖恢复了性别比。脊椎动物的遗传和环境性别决定机制并不容易解释这些结果。从生态学上讲,这种机制有利于扩大入侵新栖息地的种群,因为青蛙必须在种群之间不断迁移,以产生足够的后代,降低胚胎死亡率。
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引用次数: 2
Damage-free imaging of human chromosomes 人类染色体的无损伤成像
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.69
I. Robinson, M. Yusuf, J. Schwenke, A. Estandarte, Fucai Zhang, Gurdeep K. Bhella, Neha Parmar, J. Clark, Changyong Song, D. Nam, Gina Ratnasari, Kohei Kaneyoshi, H. Takata, K. Fukui
Microscopy methods have provided most of our knowledge to date on the structural organisation of chromosomes. Even after decades of research, the high order structure of human chromosomes is still under investigation. The new generation of X-ray sources, X-ray Free Electron Lasers (XFELs) have opened an opportunity for imaging these complex structures with higher than optical resolution but without the radiation damage that usually accompanies X-ray imaging. Here we report our first experimental steps towards imaging micron-sized human chromosomes using the SACLA XFEL facility in Japan using the MAXIC chamber. The paper highlights the sample preparation of chromosomes, staining and drying conditions used, as well as the imaging optimisations and shows our fi rst results.
显微镜方法提供了迄今为止我们对染色体结构组织的大部分知识。即使经过几十年的研究,人类染色体的高阶结构仍在研究中。新一代x射线源,x射线自由电子激光器(XFELs)为这些复杂结构的成像提供了一个机会,其分辨率高于光学分辨率,但没有通常伴随x射线成像的辐射损伤。在这里,我们报告了使用日本SACLA XFEL设备使用MAXIC室成像微米级人类染色体的第一个实验步骤。本文重点介绍了染色体的样品制备、染色和干燥条件,以及成像优化,并展示了我们的初步结果。
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引用次数: 1
Development of a robotic system interfaced to a microfl uidic device to isolate a single chromosome 一个机器人系统接口的微流体装置分离单个染色体的发展
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.73
M. Yusuf, P. Patel, Gurdeep K. Bhella, Benjamin C. Coles, K. Mir, A. Ward, S. Botchway, I. Robinson
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引用次数: 0
Comparative analysis of cytological abnormalities induced in Vicia faba L. genotypes using physical and chemical mutagenesis 物理诱变与化学诱变诱导蚕豆基因型细胞学异常的比较分析
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.47
Shahnawaz Khursheed, R. A. Laskar, A. Raina, R. Amin, Samiullah Khan
Chromosomal aberration assessment is an important index in mutation breeding for determining the mutagen potency, which helps to deduce an optimum level of mutagen dose tolerable by the species. The current experiment was carried out to study the comparative account of different concentrations of EMS (0.01%, 0.02%, 0.03% and 0.04%), gamma rays (100Gy, 200Gy, 300Gy and 400Gy) and their combinations (100 Gy + 0.01% EMS, 200 Gy + 0.02% EMS, 300 Gy + 0.03% EMS and 400 Gy + 0.04% EMS) on induced chromosomal aberrations in two varieties of Vicia faba L. viz; PRT-12 and vikrant. The control plants showed normal meiosis while the treated plants showed considerable meiotic anomaly. Percent frequency assessment showed dose dependent increase in aberrations. The different abnormalities observed were laggards, bridges, stickiness, micronuclei, disturbed polarity, cytomixis etc. Seeds treated with combination treatments showed more cytological aberrations then individual treatments of both gamma and EMS doses. The order of the most signifi cant dose from each treatment conditions (physical, chemical and combination) were 300 Gy + 0.03% EMS > 400 Gy > 0.04% EMS for var. vikrant and 400 Gy + 0.04% EMS > 0.04% EMS > 400 Gy for var. PRT-12. The comparative analysis showed higher sensitivity of variety vikrant towards the doses of mutagen used than variety PRT-12 at same dose.
染色体畸变评估是突变育种中确定诱变剂效价的重要指标,有助于推断出该物种可耐受的最佳诱变剂剂量。本试验旨在研究不同浓度电磁辐射(0.01%、0.02%、0.03%和0.04%)和γ射线(100Gy、200Gy、300Gy和400Gy)及其组合(100Gy + 0.01%电磁辐射、200Gy + 0.02%电磁辐射、300Gy + 0.03%电磁辐射和400Gy + 0.04%电磁辐射)对2个蚕豆品种染色体畸变的影响;PRT-12和维克兰特。对照植株减数分裂正常,处理植株减数分裂异常。百分比频率评估显示剂量依赖性畸变增加。不同的异常有迟滞、桥状、粘连、微核、极性紊乱、细胞分裂等。联合处理的种子比γ和EMS单独处理的种子表现出更多的细胞学畸变。各处理条件(物理、化学和组合)的最显著剂量顺序为var. vikrant为300 Gy + 0.03% EMS > 400 Gy > 0.04% EMS, var. PRT-12为400 Gy + 0.04% EMS > 0.04% EMS > 400 Gy。对比分析表明,在相同剂量下,维克兰特品种对诱变剂的敏感性高于PRT-12品种。
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引用次数: 23
Enzymatic maceration/air-drying method for chromosome observations in the young leaf of pear (Pyrus spp.) 酶浸/风干法在梨幼叶染色体观察中的应用
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.29
Masashi Yamamoto, S. Terakami, Toshiya Yamamoto
A chromosome preparation method using young leaves of pear (Pyrus spp.) was developed. Young leaves, 1-2 cm long, of grafted Japanese pear ‘Kosui’ (Pyrus pyrifolia Nakai) were used as materials. The leaves were cut into approximately 2 mm2 for enzymatic maceration/airdrying (EMA). For EMA, enzyme mixture containing 4% Cellulase Onozuka RS, 1.5% Macerozyme R200 (Yakult), 0.3% Pectolyase Y-23 (Seishin Pharmaceutical Co., Ltd.), and 1 mM EDTA, pH 4.2, at 37°C for 60-75 min was optimum for chromosome preparation because a large number of good preparations, with all 34 chromosomes relatively extended and well spread without cytoplasm, were observed. The 18S-5.8S-25S ribosomal RNA gene (rDNA) site was detected in telomeric positions of six chromosomes in fluorescent in situ hybridization (FISH). The number and positions of rDNA sites were the same as in the results using root tips as materials (Yamamoto et al. 2010, 2012). The method developed in the present study is considered to be promising for further cytogenetic studies in pear since true-to-type chromosome samples are obtained from young leaf.
研究了利用梨幼叶制备染色体的方法。以嫁接的日本梨‘小穗’(Pyrus pyrifolia Nakai)幼叶为材料,幼叶长1 ~ 2cm。将叶子切成约2 mm2用于酶浸/风干(EMA)。对于EMA,含有4%纤维素酶Onozuka RS、1.5%宏酵素R200 (Yakult)、0.3% Pectolyase Y-23 (Seishin Pharmaceutical Co., Ltd)、1 mM EDTA、pH 4.2、37℃、60-75 min的酶混合物最适合用于染色体制备,因为观察到大量良好的制剂,34条染色体均相对延长,且无细胞质分布良好。荧光原位杂交(FISH)技术在6条染色体端粒位置检测到18S-5.8S-25S核糖体RNA基因(rDNA)位点。rDNA位点的数量和位置与以根尖为材料的结果相同(Yamamoto et al. 2010, 2012)。本研究中开发的方法被认为是有希望进一步的细胞遗传学研究,因为从嫩叶中获得了真型染色体样本。
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引用次数: 1
Karyotypes of the lenok genus Brachymystax from the Amur River basin ? AgNORs are diff erent between sharp-snouted and blunt-snouted lenoks 阿穆尔河流域lenok属Brachymystax的核型研究?agnor在尖鼻和钝鼻之间是不同的
Pub Date : 2015-01-01 DOI: 10.11352/SCR.18.59
S. Frolov, H. Sakai, V. N. Frolova
Two sympatric forms of the Siberian lenok genus Brachymystax, sharp-snouted lenok (B. lenok, SS) and blunt-snouted lenok (B. tumensis is applied currently, BS), from the Amur River basin shared similar karyotype showing 2n = 90 and the formula of 8M+1SM+1SMST+9ST+27A, with NF = 110. However, AgNORs were located on one each pair of acrocentric and subtelocentric chromosomes in the SS lenok, and only one pair of subtelocentric chromosomes in the BS lenok. The observed karyotypic difference strongly supported these two forms to be different species. Variations of previously published lenok karyotypes also were discussed.
来自黑龙江流域的两种同域形态西伯利亚短鼻犀属(Brachymystax),尖鼻犀属(B. lenok, SS)和钝鼻犀属(B. tumensis, BS),核型相似,均为2n = 90,核型公式为8M+1SM+1SMST+9ST+27A, NF = 110。而在SS染色体组中,AgNORs分别位于一对远心染色体和一对亚远心染色体上,而在BS染色体组中,AgNORs仅位于一对亚远心染色体上。观察到的核型差异有力地支持这两种形式是不同的物种。还讨论了先前发表的lenok核型的变异。
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引用次数: 1
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Chromosome science
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