植物细胞遗传学实验课染色体标本建设

细胞遗传学实验课必须有一系列的实验材料作为支撑。文章通过介绍本实验室改良的植物染色体制片技术，制作了一整套完整的细胞遗传学实验教学的染色体标本，为农业院校研究生细胞遗传学实验教学提供参考。     

植物细胞减数分裂染色体制片新方法

植物细胞减数分裂染色体是遗传学实验中常用的实验标本材料，用常规制片技术制作，不易得到理想的制片效果。作者总结多年的教学工作经验，改进了传统植物细胞减数分裂染色体制片技术。新方法具有制片质量好、效率高等优点。     

单倍体育种——林木快速育种的新方法

一、林木单倍体育种的意义植物花粉(雄配子体)通过传粉作用与卵细胞(雌配子体)相结合形成受精卵。受精卵发育成为种子。种子以及由种子长成的植株的细胞中,包含着来自精、卵即父母双方的两套遗传物质——染色体,这从细胞遗传学上说是二倍体。花粉和卵细胞分别由花粉母细胞和大孢子母细胞经减数分裂而形成。它们的染色体数目为体细胞染色体的一半,也就是说,在花粉和卵细胞中只有一套染色体,因而是单倍体。花     

提高植物细胞减数分裂时染色体动态观察效果的方法

蚕豆(Vacia faba)是研究植物细胞分裂图象理想的材料,蚕豆的染色体大,它的体细胞染色体数为2n=12,易于鉴定。蚕豆在种植和管理上也较方便。因此在植物学和遗传学教学中,一般都用蚕豆花粉母细胞作为观察减数分裂染色体的动态变化材料。但由于受实验材料的取材时间与处理等因素影响,以往的实验结果都不太理想,为了能提高学生观察     

黑条小车蝗Oedaleus decorus(Gevm)精母细胞减数分裂观察研究

1、遗传学教学过程中希望学生具有遗传学的牢固基础知识。减数分裂是遗传学的三个基本原则的共同的细胞学基础。减数分裂中染色体的行为变化与生物遗传变异密切相关。熟悉减数分裂过程,掌握制片观察技术,是从事遗传育种的基本功之一。特别是细胞遗传学的优势反映在概念和技术的融合和促进。2、能否准确观察到蝗虫精母细胞的减数分裂,至关重要的是把握住蝗虫合适的发育时期:若虫末期,成虫最早期,如失之过早或偏晚都将观察不到精母细胞的减数分裂。黑条小车蝗精细管多为46条。解剖之后,凭观感可以初步判断偏晚已经退化的精细管或过早发育不良的精细管。3、黑条小车蝗精母细胞减数分裂初期Ⅰ,n=11,n=12,多为短棒状和两头带尖的端化染色体、以及少数近端着丝粒拉成括弧状的染色体图象(参见照片)。     

植物杂交概念的拓展——四论高等植物染色体杂交

在植物染色体杂交中,一个受体细胞的染色体和外源的染色体在胞内发生染色体杂交,使得分化中的受体细胞具有杂交染色体,从而发育出杂交植物.这和植物有性杂交中精子和卵子两个细胞结合不同,也和植物体细胞杂交中两个脱壁体细胞融合也不同,因此植物染色体杂交拓展了植物杂交的概念,即从两个相异细胞的融合杂交到一个细胞内两种不同染色体的融合杂交.     

甘薯细胞遗传学研究进展

指出了甘薯细胞遗传学研究仍存在的很多技术瓶颈,如功能基因定位等,总结了甘薯细胞遗传学的研究进展,包括染色体行为、杂交种染色体分析、甘薯起源与亲缘关系等.同时展望了甘薯细胞遗传学的发展和亟待加强的方向,为甘薯遗传背景的研究提供了新的方法和思路.     

罗氏沼虾的染色体研究

从罗氏沼虾处于囊胚期和原肠期的胚胎细胞中提取中期染色体进行细胞遗传学研究。染色体制片分析表明，罗氏沼虾二倍体染色体数目2N=118，但是也有一些细胞具有100～109条染色体。以外，还有一些细胞具有与单倍体染色体数相近的染色体数。     

杨山牡丹×紫牡丹杂交后代“梦幻”的细胞遗传学

进行了杨山牡丹×紫牡丹杂交后代"梦幻"体细胞的细胞遗传学研究,结果表明该品种为混倍体.对其不同染色体数目的细胞进行核型分析,核型类别分别是3B,2B,2C,与父母本有很大的差异.用植物染色体去壁低渗制片法研究了"梦幻"大孢子母细胞减数分裂双线期-后期Ⅰ的分裂过程,结果显示:所有的细胞减数分裂行为均异常,在终变期-中期Ⅰ,分别出现了多价体、单价体、二价体提前分离、染色体胶连、赤道外染色体、染色体断裂等异常现象,这些异常导致了后期Ⅰ的不等数分离,二价体或多价体不分离,染色体胶连及出现大量的染色体断片,最终不能形成正常的雌配子体.此外,在体细胞中观察到的混倍现象在生殖细胞的减数分裂过程中也得到了进一步的证实,在终变期分别出现了5_Ⅱ+断片、4_Ⅱ+3_Ⅰ,5_Ⅱ+2_Ⅰ+断片、6_Ⅱ+3_Ⅰ+断片等染色体配对构型.     

甘蓝型油菜与蓝花子属间倍半二倍体的形成及其细胞遗传学特性

对甘蓝型油菜与蓝花子的属间远缘杂交得到的杂种Ｆ＿１，经加倍后获得双二倍体植株。该植株作母本再与甘蓝型油菜杂交获得其倍半二倍体．用充气液体培养基加８－羟基喹啉预处理，对不能以根尖为材料作染色体制片的植物，以其幼嫩花蕾进行体细胞染色体制片，对甘蓝型油菜与蓝花子属间倍半二倍体的细胞遗传学特性进行了研究，该植物的染色体数目为２ｎ＝４７．大多数ＰＭＣｓ的染色体在减数分裂过程中配对不规则，并在后期Ⅱ中出现多极分裂现象．     

蝇类卵巢染色体制备的新方法

以羽化后未交尾的雌性麻蝇为活体材料,通过“Feulgan预染——显微解剖悬液铺片——Giemsa复染法”获得形态良好、分散适中、着色清晰的卵泡细胞有丝分裂中期染色体标本。本法材料性别来源明确,对蝇类性染色体鉴定的准确性有重要意义。     

三极细胞有丝分裂中期纺锤体的激光共聚焦显微镜观察

用松胞素 Ｂ（ Ｃｙｔｏｃｈａｌａｓｉｎ Ｂ, Ｃ Ｂ）处理培养的 Ｈｅｌａ 细胞,抑制胞质分裂,引起 Ｈｅｌａ 细胞发生不正常分裂,可形成多极细胞（三极、四极等）．通过荧光免疫染色法显示多极细胞有丝分裂中期的微管,使用激光共聚焦显微系统观察三极细胞纺锤体和中期染色体的空间相对关系,推测了纺锤体微管的分布与有丝分裂后期染色体分离的相关性．本方法还可用于研究有丝分裂期纺锤体微管对胞质分裂分裂沟形成的影响．     

Mitochondrial DNA repairs double-strand breaks in yeast chromosomes

The endosymbiotic theory for the origin of eukaryotic cells proposes that genetic information can be transferred from mitochondria to the nucleus of a cell, and genes that are probably of mitochondrial origin have been found in nuclear chromosomes. Occasionally, short or rearranged sequences homologous to mitochondrial DNA are seen in the chromosomes of different organisms including yeast, plants and humans. Here we report a mechanism by which fragments of mitochondrial DNA, in single or tandem array, are transferred to yeast chromosomes under natural conditions during the repair of double-strand breaks in haploid mitotic cells. These repair insertions originate from noncontiguous regions of the mitochondrial genome. Our analysis of the Saccharomyces cerevisiae mitochondrial genome indicates that the yeast nuclear genome does indeed contain several short sequences of mitochondrial origin which are similar in size and composition to those that repair double-strand breaks. These sequences are located predominantly in non-coding regions of the chromosomes, frequently in the vicinity of retrotransposon long terminal repeats, and appear as recent integration events. Thus, colonization of the yeast genome by mitochondrial DNA is an ongoing process.     

灰家鸽的核型制备及其分析

本文以灰家鸽骨髓为材料,采用增加动物骨髓细胞有丝分裂相的方法,空气干燥法制片,姬姆萨染色,获得了较理想的中期分裂相.对12只鸽子的核型观察结果表明:灰家鸽的染色体数为2n=78.文章对前10对大的染色体进行了测量和形态分析,并对家鸽与原鸽的核型以及国外报导的家鸽核型进行了初步比较,讨论了引起染色体组型差异的原因.     

Chfr defines a mitotic stress checkpoint that delays entry into metaphase

Chemicals that target microtubules induce mitotic stress by affecting several processes that occur during mitosis. These processes include separation of the centrosomes in prophase, alignment of the chromosomes on the spindle in metaphase and sister-chromatid separation in anaphase. Many human cancers are sensitive to mitotic stress. This sensitivity is being exploited for therapy and implies checkpoint defects. The known mitotic checkpoint genes, which prevent entry into anaphase when the chromosomes are not properly aligned on the mitotic spindle, are, however, rarely inactivated in human cancer. Here we describe the chfr gene, which is inactivated owing to lack of expression or by mutation in four out of eight human cancer cell lines examined. Normal primary cells and tumour cell lines that express wild-type chfr exhibited delayed entry into metaphase when centrosome separation was inhibited by mitotic stress. In contrast, the tumour cell lines that had lost chfr function entered metaphase without delay. Ectopic expression of wild-type chfr restored the cell cycle delay and increased the ability of the cells to survive mitotic stress. Thus, chfr defines a checkpoint that delays entry into metaphase in response to mitotic stress.     

利用HU和APM双阻断法诱导高频率植物根尖细胞有丝分裂同步化的研究

应用羟基脲（ｈｙｄｒｏｘｙｕｒｅａ简称ＨＵ）和甲基氨草磷（ａｍｉｐｒｏｐｈｏｓ－ｍｅｔｈｙｌ简称ＡＰＭ对小麦、大麦、黑麦、蚕豆玉米、水稻等植物进行有丝分裂同步化诱导,何使有丝分裂中期指数（ｍｅｔａｐｈａｓｅ ｉｎｄｅｘ简称ＭＩ达５０％以上,中期染色体占８０％￣９０％,有丝分裂正常,未见染色体畸变现象。     

Microtubule-motor activity of a yeast centromere-binding protein complex.

During cell division, sister chromosomes segregate from each other on a microtubule-based structure called the mitotic spindle. Proteins bind to the centromere, a region of chromosomal DNA, to form the kinetochore, which mediates chromosome attachment to the mitotic spindle microtubules. In the budding yeast Saccharomyces cerevisiae, genetic analysis has shown that the 28-basepair (bp) CDEIII region of the 125-bp centromere DNA sequence (CEN sequence) is the main region controlling chromosome segregation in vivo. Therefore it is likely that proteins binding to the CDEIII region link the centromeres to the microtubules during mitosis. A complex of proteins (CBF3) that binds specifically to the CDEIII DNA sequence has been isolated by affinity chromatography. Here we describe kinetochore function in vitro. The CBF3 complex can link DNA to microtubules, and the complex contains a minus-end-directed microtubule-based motor. We suggest that microtubule-based motors form the fundamental link between microtubules and chromosomes at mitosis.     

Movement and segregation of kinetochores experimentally detached from mammalian chromosomes

The kinetochore is a specialized structure at the centromere of eukaryotic chromosomes that attaches chromosomes to the mitotic spindle. Recently, several lines of evidence have suggested that kinetochores may have more than a passive role in the movement of chromosomes during mitosis and meiosis. Kinetochores seem to attract and 'capture' microtubules that grow from the spindle poles and microtubules may lengthen or shorten by the addition or subtraction of tubulin subunits at their kinetochore-associated ends. An attractive hypothesis is that kinetochores function as 'self-contained engines running on a microtubule track'. Here, we show that kinetochores can be experimentally detached from chromosomes when caffeine is applied to Chinese hamster ovary cells that are arrested in the G1/S phase of the cell cycle. The detached kinetochore fragments can still interact with spindle microtubules and complete all the mitotic movements in the absence of other chromosomal components. As these cells enter mitosis before DNA synthesis is completed, chromosome replication need not be a prerequisite for the pairing, alignment and segregation of kinetochores.     

大蒜根尖有丝分裂染色体标本的制作

染色体是遗传物质的载体,是基因的载体.生物体细胞的染色体数目和结构是重要的遗传标志之一,特别是在真核细胞中.因此深入认识染色体的结构和功能,对研究生物的遗传、变异、进化、细胞增殖、个体的发育和生殖过程的平衡控制都有十分重要意义.本实验以大蒜根尖为材料,介绍了大蒜根尖细胞有丝分裂标本的制作方法.该方法简单、方便,材料易得,成功率高,适用于对正常植物组织有丝分裂过程的观察.     

The inhibiting effect of 1·4 recombinant P chromosome of wheat-<Emphasis Type="Italic">Agropyron cristatum</Emphasis> addition line on the <Emphasis Type="Italic">Ph</Emphasis> gene

P chromosomes may carry a genetic system that inhibits the Ph gene in wheat. Abnormal chromosome synapsis in wheat-Agropyron cristatum addition line II-21-2 (additional 1·4 recombinant P chromosome) was observed in this study. The results of cytogenetics and Ph1 gene amplification showed that the Ph1 gene was normal and the average number of quadrivalents or hexavalents was determined to be 0.41 and 0.13, respectively, in pollen-mother cells of wheat-Agropyron cristatum addition line II-21-2. The analysis o...