Thiophene synthesis and distribution in young developing plants of Tagetes patula and Tagetes erecta

Thiophene synthesis and accumulation were investigated in organsof Tagetes patula and T. erecta. Thiophene accumulation startedrapidly in germinating seedlings of both species. Roots andhypocotyls were the major thiophene accumulating organs and5-(3-buten-1-ynyl)-2, 2-bithienyl (BBT) and 5-(4-acetoxy-1-butynyl)-2,2 -bithienyl (BBTOAc) were the major accumulated compounds.Higher thiophene concentrations were reached in Tagetes patulathan in T. erecta. Accumulation patterns for individual thiopheneswere different within organs, between organs and between bothspecies. Within hypocotyls of Tagetes patula, thiophene concentrationswere high in the epidermis and the vascular tissue and low inthe parenchymatic tissues of cortex and pith. Synthesis of thiopheneswas high in the roots and hypocotyls and very low in the leaves.Transport of thiophenes from the roots into the shoot occurred,but the rate of transport was too low to explain the high concentrationsin the hypocotyl. It is concluded that for the main part thiophenesare accumulated where they are synthesized. Key words: Tagetes, hiophenest, synthesis, accumulation, secondary metabolites     

Thiophene Content in Normal and Transformed Root Cultures of Tagetes erecta: A Comparison with Thiophene Content in Roots of Intact Plants

Transformed roots of Tagetes erecta were obtained followinginfection of stems of sterile plantlets with Agrobacterium rhizogenesstrain TR105. The thiophenes detected were 5-(4-hydroxy-l-butenyl)-2,2'-bithienyl,5-(4-acetoxy-l-butenyl)-2,2'-bithienyl, 5-(3-buten-l-enyl)-2,2',-bithienyl and 2.2': 5', 2'-terthienyl. The thiophene patternwas the same in normal root cultures and roots of the intactplant. Transformed roots showed a higher growth rate and a higherbiomass yield than normal root cultures on a hormone-free media. Key words: Transformed roots normal roots, Tagetes erecta, thiophenes, Agrobacterium rhizogenes     

Isolation and Characterization of Neurospora Mutants Affected in Invertase Synthesis

We have outlined a procedure that allows the large-scale screening of mutagenized Neurospora crassa populations for invertaseless mutants. We have isolated and characterized three mutations, inv(DBL1), inv(DBL9) and inv(DBL14), which have been mapped at or near the invertase structural gene. One of these, inv(DBL1), is particularly interesting. Our experiments indicate that the reduced level of invertase activity in the inv(DBL1)-containing cell can be explained as the result of a reduced number of normal enzyme molecules. We also show that wild-type Neurospora is able to respond rapidly to a change of medium and can dramatically increase its production of invertase within 20 min after a transfer to a carbon-free medium.     

Isolation and Identification of Pathogens Causing Marigold (Tagetes erecta L.) Black Spot in Beijing,China

Black spot leads to great marigold losses worldwide. The disease is characterized by black spots on leaves and stems in its early stages, and the whole plant has black rot at the advanced stage. In this report, 6 of 217 Alternaria strains isolated from lesions of marigold plants in Beijing were randomly selected. The morphological characteristics and a pathogenic tree based on two protein‐coding genes (gpd and alt a 1) indicated that Alternaria tagetica is the causal agent of marigold black spot in Beijing. All six Alternaria strains could successfully re‐infect marigold, but they could not infect carrot or zinnia by either spore spray in a greenhouse or planting experiments in the epidemic area. This is the first report of the A. tagetica pathogen being isolated from marigold in Beijing.     

Thiophene production in transformed roots cultures of Tagetes filifolia

Transformed root cultures of Tagetes filifolia were established by infection with Agrobacterium rhizogenes LBA 9402. Several clones were obtained with different growth index, content of thiophenes and pattern. The isolated clones showed considerable variations in total thiophene content (27 to 11764 g/g fresh weight) but all had similar patterns of the different thiophenes. Only one clone excreted thiophenes (about 80% of total amount). © Rapid Science Ltd. 1998     

Effects of vesicular-arbuscular mycorrhiza on Tagetes erecta and Zinnia elegans

 Seeds of Tagetes erecta and Zinnia elegans were planted in soil inoculated with the vesicular-arbuscular mycorrhizal fungus Glomus etunicatum. This procedure produced positive effects on both Tagetes and Zinnia compared with the control: faster flowering and an increased number of flowers. Shoot height, shoot and root dry weights and percentage infected root length were also measured. The reactions of these plants to mycorrhizal infection were shown to be independent of changes in the phosphorus, potassium and sodium contents of the plants. Accepted: 1 August 1995     

Isolation and Characterization of Aminopeptidase-Deficient Lactobacillus bulgaricus Mutants

Lactobacillus bulgaricus CNRZ 397 is able to hydrolyze many amino-acyl- and dipeptidyl-β-naphthylamides. Analysis of heat inactivation kinetics, protease inhibitor effects, and the subcellular location of aminopeptidase (AP) activities from the parental strain and mutant derivatives dificient in alanyl- or leucyl-β-naphthylamide hydrolysis pointed out the existence of four APs. All mutants isolated were totally deficient in AP II, a cell wall metallo-enzyme with a broad substrate specificity but that is specifically responsible for lysyl-AP activity and is characterized by a molecular mass of 95,000 daltons. AP I and AP III are cytoplasmic enzymes that exhibit arginyl-AP activity; both enzymes are inducible during growth in rich peptide MRS medium (Difco Laboratories, Detroit, Mich.). The existence of a fourth AP (AP IV) that is involved in leucyl-AP activity was suggested. Moreover, we showed that X-prolyl-dipeptidyl-AP activity, which was not catalyzed by an AP, involved an enzyme(s) that is controlled by a regulatory mechanism that is common to that of AP II.     

Isolation and Characterization of Sendai Virus Temperature-Sensitive Mutants

Ten temperature-sensitive mutants of Sendai virus, a paramyxovirus, were isolated and partially characterized. The mutants replicated in chicken embryo lung cells at 30 C, but not at 38 C; wild-type virus grew equally well at both temperatures. Complementation tests divided the mutants into seven groups. Six groups synthesized neither infectious virus nor RNA when incubated at 38 C from the beginning of infection. Temperature shift-up experiments demonstrated that three of these complementation groups were blocked in early steps required for RNA synthesis, but these gene functions were not needed throughout the replicative cycle. In contrast, the other three RNA-negative complementation groups were defective throughout the replicative cycle in functions required for virus-specific RNA synthesis. Only one mutant, which complemented all of the above, synthesized RNA but not infectious virus when placed at 38 C; the hemagglutinin of this mutant functioned only at the permissive temperature.     

Agrobacterium-mediated transient transformation of marigold (Tagetes erecta)

Hypocotyls, roots, leaf sections and shoot tips from Tagetes erecta plantlets were inoculated with Agrobacterium tumefaciens, harboring the binary vector pCAMBIA2301, containing the β-glucuronidase gene. Histochemical GUS assays of infected tissues showed transient gus gene expression after 3 days.     

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.     

缺硫对万寿菊镉积累的影响

采用营养液培养法,研究了缺硫对万寿菊(Tagetes erecta)各器官镉积累的影响,以及巯基化合物和有机酸等在各器官镉积累中的作用.结果表明:0.1 mg·L-1镉处理7 d,缺硫处理显著降低了植物根、茎和叶中的镉含量,较对照分别降低了47%、38%和61%;缺硫对植物体内半胱氨酸、γ-谷氨酰半胱氨酸、谷胱甘肽和有机酸的合成没有显著影响,而且3种巯基化合物的含量都很低;缺硫植物根部合成的植物络合素的量显著降低,这可能是植物体内镉积累减少的主要原因;在植物的各器官中,叶片中的有机酸含量最高,而在叶片中未检测到植物络合素,由此可推断,叶片中积累的镉主要与有机酸结合;缺硫影响了根部植物络合素的合成,进而降低了植物体内镉的积累,而缺硫没有影响植物体内有机酸的合成,有机酸可能仅是参与植物组织中与镉的络合作用.     

Isolation and Characterization of NaCl-Sensitive Mutants of Caulobacter crescentus

An attempt to characterize Caulobacter crescentus genes important for the response to high concentrations of NaCl was initiated by the isolation of mutants defective in survival in the presence of 85 mM NaCl. A transposon Tn5 library was screened, and five strains which contained different genes disrupted by the transposon were isolated. Three of the mutants had the Tn5 in genes involved in lipopolysaccharide biosynthesis, one had the Tn5 in the nhaA gene, which encodes a Na⁺/H⁺ antiporter, and one had the Tn5 in the ppiD gene, which encodes a peptidyl-prolyl cis-trans isomerase. All the mutant strains showed severe growth arrest in the presence of 85 mM NaCl, but only the nhaA mutant showed decreased viability under these conditions. All the mutants except the nhaA mutant showed a slightly reduced viability in the presence of 40 mM KCl, but all the strains showed a more severe reduction in viability in the presence of 150 mM sucrose, suggesting that they are defective in responding to osmotic shock. The promoter regions of each disrupted gene were cloned in lacZ reporter vectors, and the pattern of expression in response to NaCl and sucrose was determined; this showed that both agents induced ppiD and nhaA gene expression but did not induce the other genes. Furthermore, the ppiD gene was not induced by heat shock, indicating that it does not belong to the σ³² regulon, as opposed to what was observed for its Escherichia coli homolog.     

Isolation and Characterization of Sex-Linked Female-Sterile Mutants in DROSOPHILA MELANOGASTER

The purpose of the experiments described was to identify X chromosome genes functioning mainly or exclusively during oogenesis. Two mutagenesis experiments were carried out with ethyl methane sulfonate. Following treatment inducing 60% lethals, 9% of the treated X chromosomes carried a female sterility mutation which did not otherwise seriously affect viability. Among —95 isolated mutants, 19 were heat-sensitive and 5 cold-sensitive. The mutants have been classified as follows: I (16 mutants; 12 complementation groups): the females laid few or no eggs; the defect concerned either ovulation or oogenesis. II (37 mutants; 18 complementation groups): the female laid morphologically abnormal eggs, often with increased membrane permeability. III A (13 mutants; at least 8 complementation groups): the homozygous females were sterile if mated to mutant males; their progeny (homo- and hemizygous) died at a late embryonic stage (11 mutants), at the larval stage (1 mutant) or at the pupal stage (1 mutant). However fertility was partly restored by breeding to wild-type males as shown by survival of some heterozygous descendants. III B (29 mutants; 22 complementation groups): the fertility of the females was not restored by breeding to a wild-type male. Most of the eggs of 13 of the mutants died at a late stage of embryogenesis. The eggs of the others ceased development earlier or, perhaps, remained unfertilized. The distribution of the number of mutants per complementation group led to an estimation of a total of about 150 X-linked genes involved in female fertility. The females of three mutants, heat-sensitive and totally sterile at 29°, produced at a lower temperature descendants morphologically abnormal or deprived of germ cells. Three other mutants not described in detail showed a reduction in female fertility with many descendants lacking germ cells. A desirable mutant which was not recovered was one with normal fertile females producing descendants which, regardless of their genotype, bore specific morphological abnormalities. The value of the mutants isolated for analysis of the complex processes leading to egg formation and initiation of development is discussed.     

Isolation and Characterization of Light-Insensitive Mutants of NEUROSPORA CRASSA

As part of a genetic analysis of blue light photoreception in Neurospora, three mutants were isolated that do not exhibit photosuppression of circadian conidiation, i.e., they show periodic conidiation in constant light. The mutations have been given the designations lis-1, lis-2 and lis-3 ("light insensitive"). The three mutations segregate as single nuclear genes, are nonallelic and are recessive to wild type in heterokaryon tests. The linkage groups of the mutations are as follows: lis-1, I; lis-2, VI; and lis-3, V. The light -insensitive phenotype of the mutants is restricted to the photosuppression response; other responses such as photoinduced phase shifting of the conidiation rhythm and photoinduced carotenogenesis are not altered. The physiological or biochemical defects of the mutants have not been established, but they are not similar to previous reported cases (i.e., rib and poky) in which a reduction in light sensitivity has been observed.     

Isolation and Characterization of Mms-Sensitive Mutants of SACCHAROMYCES CEREVISIAE

We have isolated mutants sensitive to methyl methanesulfonate (MMS) in Saccharomyces cerevisiae. Alleles of rad1, rad4, rad52, rad55 and rad57 were found amoung these mms mutants. Twenty-nine of the mms mutants which complement the existing radiation-sensitive (rad and rev) mutants belong to 22 new complementation groups. Mutants from five complementation groups are sensitive only to MMS. Mutants of 11 complementation groups are sensitive to UV or X rays in addition to MMS, mutants of six complementation groups are sensitive to all three agents. The cross-sensitivities of these mms mutants to UV and X rays are discussed in terms of their possible involvement in DNA repair. Sporulation is reduced or absent in homozygous diploids of mms mutants from nine complementation groups.