Cytogenetic Analysis of Cotton Monosomics

Eleven monosomics in cotton that were obtained in the progenies of three disomic desynaptic plants were cytologically characterized. The transmission of the monosomes in progeny was shown in the 26 monosomic plants. In 23 plants the frequency of monosomics was ranged between 14.29 and 41.67 %. Three monosomics usually occurred in much lower frequencies (from 3.03 to 5.00 %). Various transmission rates indirectly pointed out different monosomes as a specific chromosomes of cotton genome. Three telochromosomes and one isochromosome were isolated from the progenies of the four monosomics. Using translocation test it was recovered that seven monosomes of different monosomics are homologous to one of the chromosomes of six translocation lines of our collection.     

Chromosomal Banding Analysis of Monosomics in Wheat

N-banding analysis has been used to identify the univalents of all 21 monosomics at diakinesis or metaphase Ⅰ. The univalents of nine wheat monosomics which are monosomic lB to 7B, 4A and 7A have shown distinctive N-banding patterns. These banding patterns appear to be identical in meiotic and mitotic chromosomes. The method is simple and speedy. The research probably provides a new way for cytological identification of monosomics in wheat and offers a technique for genome analysis of hybrids in wheat.     

Cyto-Chromosomal Analysis of Developing Wheat Monosomics

In order to achieve the aim of advanced breeding program with the definite direction, it is necessary for us to develop the monosomic lines used for the wheat breeding programs in China. We fixed the wheat ears at appropriate stage in Carnoy’s fluid, and stained with acetocarmine in every generation from the different crosses mentioned above. According to their karyotypes of metaphase 1, the monosomics, normal bodies, monotelosomics, ditelosomics and allotypic bivalents were identified (Plate Ⅰ, 1–8). In the process of developing monosomic lines “Beijing Red No.1”, some monosomic lines such as 5A’s and 4D’s, can be directly proved by their phenotypes, other lines of monosomic 1B, 5B add 6B can also be directly proved to be true by their typical chromosomal morphology. In order to check the accuracy of chromosomal orders of monosomic lines, we tested all 21 monosomic lines of “Beijing Red No.1” by means of telosomic testing. At the same time we tested the origirnal monosomics of “Chinese Spring” as a check. In the F1’s of test crosses, those showing 20 bivalents and one monoelemic (20”+t’) were proved to be right. Whereas those showing 19 bivalents, 1 univalent and 1 allotypic bivalent (19”+1’+1’t’) were proved to be wrong. The karyotypes of F1’s from the test crosses for “Beijing Red No.1” can be verified by compairing with that of the check. During some years, we have examined 500 F1 plants of test crosses for monosomic lines of “Beijing Red No.1”, and some what less plants for monosomic lines of “Chinese Spring”. The number of observed cells usually was 100–200, the least was 40 and the most was 600. As the result, all F1's of test crosses showed accurate karyotypes. Besides detemning the F1 karyotypes of test crosses, we also analysed and compared their phenotypes with each other (photograph 9–12). According to the pbenotypes caused by the chang in chromosome number, structure and gene dosage, not only we can check the accuracy of testing result, but also locate the genes controlling some characters on the chromosomes or chromosomal arms.     

Isolation and Characterization of Amber Suppressors in Yeast

Nonsense suppressors were obtained in a haploid yeast strain containing eight nutritional mutations, that are assumed to be amber or ochre, and the cyc1-179 amber mutation that has a UAG codon corresponding to position 9 in iso-1-cytochrome c. Previous studies established that the biosynthesis and function of iso-1-cytochrome c is compatible with replacements at position 9 of amino acids having widely different structures (Stewart and Sherman 1972). UV-induced revertants, selected on media requiring the reversion of one or two of the amber nutritional markers, were presumed to contain a suppressor if there was the unselected reversion of at least one other marker. The 1088 suppressors that were isolated could be divided into 78 phenotypic classes. Only 43 suppressors of three classes caused the production of more than 50% of the normal amount of iso-1-cytochrome c in the cyc1-179 strain. Genetic analyses indicated that all of these highly efficient amber suppressors are allelic to one or another of the eight suppressors which cause the insertion of tyrosine at ochre (UAA) codons (Gilmore, Stewart and Sherman 1971). Furthermore, only tyrosine has been identified at position 9 in iso-1-cytochrome c in cyc1-179 strains suppressed with these efficient amber suppressors.     

Monosomics in common bean,Phaseolus vulgaris

Summary Two monosomics of Phaseolus vulgaris (2n = 22) were found among selfed progeny of plants treated with colchicine. The monosomic chromosomes involved were identified as chromosomes H and J according to the previously suggested Giemsa karyotype. Both monosomic plants had slower growth rate and smaller size as compared with their respective euploid sibs. However, no apparent morphological characteristics distinguished the two monosomics. The frequencies of transmission through selfing of monosomics H and J were 9% and 10 % respectively.     

Isolation and Identification of a Sexual Hormone in Yeast

5-Fluorotryptophan (5FT), indolmycin (IM), 4-fluorotryptophan and 7-azatryptophan were found on screening to be tryptophan antagonists among various chemically synthesized and naturally occurring tryptophan analogues for the isolation of l-tryptophan (l-Trp) producing mutants of Bacillus subtilis K.

From among 5FT resistant mutants, potent l-Trp producers were obtained using an improved isolation medium. Growth of the isolated 5FT-resistant l-Trp producer, AJ 11709, was inhibited by IM. From among 5FT and IM resistant mutants, the best strain, AJ 11979, which produced 9.0 g/liter of l-Trp from 13% glucose on 120hr cultivation, was selected.     

高产虾青素红发夫酵母选育研究进展

虾青素是 60 0多种类胡萝卜素中的一种 ,具有抗氧化 ,抗肿癌和增强免疫力等许多重要的生理和生物学功能 ,在水产养殖、食品和医药等领域应用前景广阔。综述了虾青素的生物来源、生物转化途径以及高产虾青素红发夫酵母菌株的选育。     

一种简便的适用于酵母双杂交系统的酵母质粒提取方法

目的:建立一种适用于酵母双杂交系统的简便快捷的酵母质粒提取方法。方法:以酿酒酵母为供试材料,用玻璃珠振荡法破除酵母细胞壁,提取酵母总DNA,最后通过电转化大肠杆菌DH10B获得目的质粒。结果:粗提得到的质粒可直接转化DH10B,作为模板用于PCR分析及酵母双杂交后续的序列分析等,大大降低了工作量。结论:该方法简便快捷,经济实用,降低了成本,提高了效率,可以作为一种实验室酵母质粒提取方法。     

Isolation and Characterization of a Thermotolerant Methanol-Utilizing Yeast

A yeast capable of growth on methanol as its sole carbon-energy source was isoalted from soil samples and identified as a strain of Hansenula polymorpha. A continuous enrichment culture at 37 C with a simple mineral salts medium was used to select this organism. The isolate, designated DL-1, has a maximal specific growth rate of 0.22 per h, at pH 4.5 to 5.5 and temperatures of 37 to 42 C, in simple mineral salts medium with methanol (0.5%), biotin, and thiamine. Growth occurred in a chemostat at temperatures up to 50 C, with strong growth at 45 C. The maximal growth yield of the yeast on methanol was 0.36 g of dry cell weight per g of methanol, and the yield on oxygen was 0.37 g of dry cell weight per g of O(2). Protein content of the isolate is 46%, and total nucleic acid content varies from 5.0 to 7.0% with increasing growth rate from 0.08 to 0.20 per h. The amino acid profile of this yeast protein indicates that it could serve as a good source of food protein. Feeding studies with rats show the yeast to have no toxic effects.     

Isolation and Characterization of Chromosome-Gain and Increase-in-Ploidy Mutants in Yeast

We have developed a colony papillation assay for monitoring the copy number of genetically marked chromosomes II and III in Saccharomyces cerevisiae. The unique feature of this assay is that it allows detection of a gain of the marked chromosomes even if there is a gain of the entire set of chromosomes (increase-in-ploidy). This assay was used to screen for chromosome-gain or increase-in-ploidy mutants. Five complementation groups have been defined for recessive mutations that confer an increase-in-ploidy (ipl) phenotype, which, in each case, cosegregates with a temperature-sensitive growth phenotype. Four new alleles of CDC31, which is required for spindle pole body duplication, were also recovered from this screen. Temperature-shift experiments with ipl1 cells show that they suffer severe nondisjunction at 37°. Similar experiments with ipl2 cells show that they gain entire sets of chromosomes and become arrested as unbudded cells at 37°. Molecular cloning and genetic mapping show that IPL1 is a newly identified gene, whereas IPL2 is allelic to BEM2, which is required for normal bud growth.     

Yeast Membrane Vesicles: Isolation and General Characteristics

Yeast membrane vesicles are formed when packed yeast are ground manually in a porcelain mortar and pestle with glass beads (0.2 mm diameter). These vesicles can be separated from the other components of the grinding mixture by a combination of centrifugation steps and elution from a column of the same glass beads (0.2 mm diameter). Isolated vesicles are osmotically sensitive, contain cytoplasmic components, and have energy-independent transport function. They are unable to metabolize glucose, but have respiratory function which is thought to be associated with intravesicular mitochondria. Invertase and oligomycin-insensitive adenosine triphosphatase are present in lysed vesicle preparations, and the appropriateness of these enzyme activities as membrane markers is discussed.     

Starvation-Associated Genome Restructuring Can Lead to Reproductive Isolation in Yeast

Knowledge of the mechanisms that lead to reproductive isolation is essential for understanding population structure and speciation. While several models have been advanced to explain post-mating reproductive isolation, experimental data supporting most are indirect. Laboratory investigations of this phenomenon are typically carried out under benign conditions, which result in low rates of genetic change unlikely to initiate reproductive isolation. Previously, we described an experimental system using the yeast Saccharomyces cerevisiae where starvation served as a proxy to any stress that decreases reproduction and/or survivorship. We showed that novel lineages with restructured genomes quickly emerged in starved populations, and that these survivors were more fit than their ancestors when re-starved. Here we show that certain yeast lineages that survive starvation have become reproductively isolated from their ancestor. We further demonstrate that reproductive isolation arises from genomic rearrangements, whose frequency in starving yeast is several orders of magnitude greater than an unstarved control. By contrast, the frequency of point mutations is less than 2-fold greater. In a particular case, we observe that a starved lineage becomes reproductively isolated as a direct result of the stress-related accumulation of a single chromosome. We recapitulate this result by demonstrating that introducing an extra copy of one or several chromosomes into naïve, i.e. unstarved, yeast significantly diminishes their fertility. This type of reproductive barrier, whether arising spontaneously or via genetic manipulation, can be removed by making a lineage euploid for the altered chromosomes. Our model provides direct genetic evidence that reproductive isolation can arise frequently in stressed populations via genome restructuring without the precondition of geographic isolation.     

从老抽酱醪中分离耐盐性酵母的研究

以老抽酱醪为实验材料进行耐盐性酵母菌种分离,并做菌种鉴定。分析了在不同盐度条件下耐盐性酵母菌的生长情况和生长过程中培养基总糖的消耗,可以发现实验得到的酵母在22％（质量与体积}E）盐度下依然能够良好生长。结果表明,实验分离出的No．2菌在同级盐度的条件下的生长量要明显高于No．1菌,但在乙醇产率方面,两株菌在相同的含盐量为16％（质量与体积比）的麦芽汁培养基中发酵8d,No．1菌的乙醇产率为3．1％（体积比）,No．2菌的乙醇产率2．9％（体积比）。     

Isolation and Characterization of Omnipotent Suppressors in the Yeast Saccharomyces Cerevisiae

Approximately 290 omnipotent suppressors, which enhance translational misreading, were isolated in strains of the yeast Saccharomyces cerevisiae containing the psi+ extrachromosomal determinant. The suppressors could be assigned to 8 classes by their pattern of suppression of five nutritional markers. The suppressors were further distinguished by differences in growth on paromomycin medium, hypertonic medium, low temperatures (10 degrees), nonfermentable carbon sources, alpha-aminoadipic acid medium, and by their dominance and recessiveness. Genetic analysis of 12 representative suppressors resulted in the assignment of these suppressors to 6 different loci, including the three previously described loci SUP35 (chromosome IV), SUP45 (chromosome II) and SUP46 (chromosome II), as well as three new loci SUP42 (chromosome IV), SUP43 (chromosome XV) and SUP44 (chromosome VII). Suppressors belonging to the same locus had a wide range of different phenotypes. Differences between alleles of the same locus and similarities between alleles of different loci suggest that the omnipotent suppressors encode proteins that effect different functions and that altered forms of each of the proteins can effect the same function.     

Presumed Sexual Isolation in Yeast Populations during Production of Sherrylike Wine

The three stages in wine production in the Montilla-Moriles region of Spain are (i) fermentation, (ii) film formation in vessels, and (iii) aging, all of which selected three yeast populations. All isolates from the three stages were classified as Saccharomyces cerevisiae. The distribution of characteristics of sugar fermentation (sucrose, galactose, maltose), Li tolerance, and Cu tolerance was different in the strains isolated in the three stages. This finding suggests that sexual isolation in the yeast populations prevents the random distribution of taxonomic characters.     

两相分配法制备南极红酵母质膜

用葡聚糖T-500(Dextran T-500)和聚乙二醇(PEG-3350)两相体系制备南极红酵母(Rhodotorula sp.)菌株 NJ298的质膜。首先在2 mmol/L KCl浓度下, 选用5种不同的聚合物浓度(5.6%、5.8%、6.0%、6.2%、6.4 %, W/W), 研究了NJ298质膜在两相体系中的分配情况, 在此基础上进一步研究了KCl浓度(2 mmol/L、4 mmol/L、6 mmol/L、8 mmol/L、10 mmol/L)对NJ298质膜的纯度及得率的影响。结果表明, 选用6.0%聚合物浓度, 4 mmol/L KCl的两相分配体系, 分离3次可得到相对纯度在78.2%的南极红酵母质膜组分, 标志酶鉴定及磷钨酸染色电镜检测均表明获得了高纯度密实的正向型的质膜囊泡。这为进一步研究该菌株的南极极端环境适应机制奠定了基础。     

两相分配法制备南极红酵母质膜

用葡聚糖 T-500(Dextran T-500)和聚乙二醇(PEG-3350)两相体系制备南极红酵母(Rhodotorula sp.)菌株 NJ298 的质膜.首先在 2 mmol/L KCl 浓度下,选用5种不同的聚合物浓度(5.6%、5.8%、6.0%、6.2%、6.4%,W/W),研究了 NJ298 质膜在两相体系中的分配情况,在此基础上进一步研究了 KCl 浓度(2 mmol/L、4 mmol/L、6 mmol/L、8 mmol/L、10 mmol/L)对 NJ298 质膜的纯度及得率的影响.结果表明,选用6.0%聚合物浓度,4 mmol/LKCl 的两相分配体系,分离3次可得到相对纯度在 78.2%的南极红酵母质膜组分,标志酶鉴定及磷钨酸染色电镜检测均表明获得了高纯度密实的正向型的质膜囊泡.这为进一步研究该菌株的南极极端环境适应机制奠定了基础.