Rhodamine-phalloidin and anti-tubulin antibody staining of spindle fibres that were irradiated with an ultraviolet microbeam

Summary We irradiated chromosomal spindle fibres in crane-fly spermatocytes with an ultraviolet microbeam of 270 nm wavelength light with total energies near those that cause actin filaments in myofibrils to depolymerize; after irradiation we stained the cells with rhodamine-labelled phalloidin and with anti-tubulin antibodies. In some cells, the irradiation reduced both phalloidin and tubulin staining of the chromosomal spindle fibres; in other cells, the irradiations reduced phalloidin staining but not tubulin staining; in yet other cells, the irradiations reduced tubulin staining but not phalloidin staining. In all irradiated cells in which phalloidin staining was reduced in the irradiated areas phalloidin staining also was reduced poleward from the irradiated areas. These results show that phalloidin staining of chromosomal spindle fibres is not dependent on the presence of kinetochore microtubules, and, therefore, that actin filaments are present in the spindle fibres in vivo. We suggest that actin filaments present in spindle fibres in vivo may be involved in causing chromosome movements during anaphase.     

PET1402, a nuclear gene required for proteolytic processing of cytochrome oxidase subunit 2 in yeast

The nuclear mutation pet ts1402 prevents proteolytic processing of the precursor of cytochrome oxidase subunit 2 (cox2) in Saccharomyces cerevisiae. The structural gene PET1402 was isolated by genetic complementation of the temperature-sensitive mutation. DNA sequence analysis identified a 1206-bp open reading frame, which is located 215 by upstream of the PET122 gene. The DNA sequence of PET1402 predicts a hydrophobic, integral membrane protein with four transmembrane segments and a typical mitochondrial targeting sequence. Weak sequence similarity was found to two bacterial proteins of unknown function. Haploid cells containing a null allelle of PET1402 are respiratory deficient.     

Role of the Cys 2-Cys 10 disulfide bond for the structure, stability, and folding kinetics of ribonuclease T1.

The Cys 2-Cys 10 disulfide bond in ribonuclease T1 was broken by substituting Cys 2 and Cys 10 by Ser and Asn, respectively, as present in ribonuclease F1. This C2S/C10N variant resembles the wild-type protein in structure and in catalytic activity. Minor structural changes were observed by 2-dimensional NMR in the local environment of the substituted amino acids only. The thermodynamic stability of ribonuclease T1 is strongly reduced by breaking the Cys 2-Cys 10 bond, and the free energy of denaturation is decreased by about 10 kJ/mol. The folding mechanism is not affected, and the trans to cis isomerizations of Pro 39 and Pro 55 are still the rate-limiting steps of the folding process. The differences in the time courses of unfolding and refolding are correlated with the decrease in stability: the folding kinetics of the wild-type protein and the C2S/C10N variant become indistinguishable when they are compared under conditions of identical stability. Apparently, the Cys 2-Cys 10 disulfide bond is important for the stability but not for the folding mechanism of ribonuclease T1. The breaking of this bond has the same effect on stability and folding kinetics as adding 1 M guanidinium chloride to the wild-type protein.     

大蒜中期染色体内的核糖核蛋白物质

研究了大蒜（Ａｌｌｉｕｍ ｓａｔｉｖｕｍ Ｌ．）中期染色体的超微结构和ＲＮＰ物质。常规染色表明,大部分染色体内部有低电子密度区,有的染色体中低电子密度区域较大而似孔洞。银染结果也证明了有大小不等的孔洞存在。Ｂｅｒｎｈａｒｄ 染色显示,在染色体周边和染色体内部都有ＲＮＰ分布。用ＮａＯＨ 处理证明了Ｂｅｒｎｈａｒｄ 染色法所显示的深染区确实含有ＲＮＡ。ＲＮＰ量的多少与ＥＤＴＡ 的分化时间呈负相关     

Cytogeography and meiotic chromosome configurations of six intraspecific aneuploids ofCarex conica Boott (Cyperaceae) in Japan

Chromosome numbers were determined for 340 plants ofCarex conica from 83 populations in Japan. Six aneuploids, 2n=32, 33, 34, 36, 37 and 38, were found. Plants with even diploid chromosome numbers 2n=32, 34, and 36 were the most common and had different geographical distributions. Individuals with 2n=32 were from islands in the Seto Inland Sea and nearby coastal areas of the Chugoku District of Honshu; those with 2n=34 were from the Kanto, Chubu and Kinki Districts of Honshu; those with 2n=36 were from the mountainous areas of Chugoku, Shikoku and Kyushu Districts. Canonical discriminant analysis of 17 morphological characters demonstrated that the plants with 2n=32 were clearly distinct from those with 2n=34 or 36. All four aneuploids with even chromosome numbers showed normal bivalent pairing at meiotic metaphase I and probably represent cytogenetically stable cytodemes. Plants with 2n=33 had one heteromorphic trivalent and 15 bivalents, indicating a structural mutation. At mitotic metaphase I, one chromosome was markedly larger than the others, suggesting that the 2n=33 plants arose from 2n=34 plants by fusion of two chromsomes. The plant with 2n=37 was intermediate in morphology betweenCarex conica (2n=36) andC. morrowii (2n=38) and probably originated as an interspecific hybrid between these species.     

Determination of the size of the Azotobacter vinelandii chromosome

The chromosome of Azotobacter vinelandii UW was digested separately with the rape cutter restriction endonucleases Swal (5-ATTTAAAT), PmeI (5GTTTAAAC) and Pacl (5-TTAATTAA) and the products were separated by pulsed-field gel electrophoresis. The size of the chromosome was determined to be approximately 4.5 megabase pairs (Mb) based on the sum of the sizes of the restriction fragments. This is almost the same as the size of the chromosome of Escherichia coli. The inability of the undigested DNA to enter the gel has led us to infer that the chromosome is circular.     

A self-fertile trigeneric hybrid,Triticum aestivum ×Agropyron michnoi ×Secale cereale

Trigeneric hybrids between the (Triticum aestivum ×Agropyron michnoi) F₁ (CM, 2n=5x=35; ABDPP) and two winter rye (Secale cereale L., 2n=2x=14; RR) cultivars, Wugong 774 and AR-132, were synthesized. Such trigeneric hybrids could be used to transfer resistance genes for powdery mildew from rye to CM and subsequently to common wheat and to identify (1) the effects of the P genome ofAgropyron on the self-fertility of the hybrids and (2) the differences in genetic background between rye cultivars with marked differences in pollinating habit. The trigeneric hybrids varied widely in morphology and showed a high level of resistance to such diseases as barley yellow dwarf virus (BYDV), stripe rust, leaf rust, stem rust, and powdery mildew. Selfed and many backcross derivatives were obtained from the trigeneric hybrids. The results indicated that rye cvs Wugong 774 and AR132 arose from different gene pools and that the P genome ofAgropyron carries gene(s) responsible for chromosome segregation, leading to functional gamete formation and self-fertility of the hybrids. The F₂ and BC₁ plants could be obtained in two ways — fusion of the unreduced gametes and the assumed apomixis of unreduced female gametes in the trigeneric hybrid plant II-4 — which indicates that this trigeneric hybrid may be a special genetic stock. Chromosome pairing in the trigeneric hybrids and ways of producing wheat/rye and wheat/Agropyron translocations are discussed.     

穗花杉染色体的研究

穗花杉为国家三级重点保护的珍稀濒危植物,雌雄异株。根尖细胞染色体分析表明：雌株和雄株的染色体数目为２ｎ＝４０,其中第１对和第２对为中部着丝粒染色体,第３－２０对为端部着丝粒染色体。核型为２ｎ＝４０＝４ｍ＋３６Ｔ。雌雄株除第２对的长度稍有差异外,其余各对的相对长度和臂比都较近似,可能尚无性染色体分化。Ｇｉｅｍｓａ Ｃ带显示,间期核有３个较大的染色中心,最长的３条染色体的中央缢痕有深染色带纹,可能是着