Teosinte crossing barrier1, a locus governing hybridization of teosinte with maize

A teosinte gene or gene cluster, Teosinte crossing barrier1 (Tcb1), that restricts crossability with maize mapped 6 centiMorgans distal to sugary-1 on chromosome 4. Tcb1 is loosely linked with the gametophyte-1 locus whose Ga1-s allele, present in many popcorns, confers nonreceptivity to the pollen of other maize varieties (ga1). Full-strength Tcb1 (positive modifiers present) was nonreceptive to Ga1-s as well as to ga1 pollen. Attenuated Tcb1 (positive modifiers absent) was detectably more receptive to Ga1-s than to ga1, suggesting cross recognition between the two systems of incompatibility. Reciprocally, homozygous Ga1-s was unreceptive both to Tcb1 and tcb1 pollen, but heterozygous Ga1-s/ga1 plants were somewhat more receptive to Tcb1 than to tcb1. Discrimination by Tcb1/− females against tcb1 pollen is prezygotic, accomplished without the loss of viable ovules. When introduced into maize, Tcb1 incompatibility may be useful for isolating one category of commercial varieties from another. Received: 25 September 2000 / Accepted: 24 November 2000     

Genetic interaction of DED1 encoding a putative ATP-dependent RNA helicase with SRM1 encoding a mammalian RCC1 homolog in Saccharomyces cerevisiae

 The Saccharomyces cerevisiae temperature-sensitive mutants srm1-1, mtr1-2 and prp20-1 carry alleles of a gene encoding a homolog of mammalian RCC1. In order to identify a protein interacting with RCC1, a series of suppressors of the srm1-1 mutation were isolated as cold-sensitive mutants and one of the mutants, designated ded1-21, was found to be defective in the DED1 gene. The double mutant, srm1-1 ded1-21, could grow at 35° C, but not at 37° C. A revertant of srm1-1 ded1-21 that became able to grow at 37° C acquired another mutation in the SRM1 gene, indicating the tight relationship between SRM1 and DED1. In all the rcc1 ^- strains examined, the amount of mutated SRM1 proteins was reduced or not detectable at the nonpermissive temperature. While mutated SRM1 protein was stabilized in all of the rcc1 ^- strains by the ded1-21 mutation, the ded1-21 mutation suppressed both srm1-1 and mtr1-2, but not the prp20-1 mutation, contrary to the previous finding that overproduction of the S. cerevisiae Ran homolog GSP1 suppresses prp20-1, but not srm1-1 or mtr1-2. Received: 20 March 1996/Accepted: 1 July 1996     

Synthesis of Several Fructo-oligosaccharides by Asparagus Fructosyltransferases

Nine fructo-oligosaccharides, synthesized in vitro from sucrose by an enzyme preparation from asparagus roots, were isolated and their structures were elucidated to be 1^F (1-β-fructofuranosyl)_n sucrose [n = 1 (1-kestose), 2 (nystose) and 3], 6^G (1-β-fructofuranosyl)_n sucrose [n=1 (neokestose), 2 and 3] and 1^F (1-β-fructofuranosyl)_m-6^G (1-β-fructofuranosyl)_n sucrose [m=1, n=1; m=2, n =1; and m =1, n=2]. These saccharides are all known to occur naturally in asparagus roots, but 6^G (1-β-fructofuranosyl)₃ sucrose and 1^F (1-β-fructofuranosyl)_m-6^G-(1-β-fructofuranosyl)_n sucrose (m=1, n =1; and m=1, n=2) were the first saccharides enzymatically synthesized in vitro. Also three types of fructosyltransferases were presumed to be involved in the biosynthesis of these oligosaccharides in asparagus roots.     

Expression of the SNF1 gene from Candida tropicalis is required for growth on various carbon sources, including glucose

SNF1 of Saccharomyces cerevisiae is an essential gene for the derepression of glucose repression. A homolog of SNF1 (CtSNF1) was isolated from an n-alkane-assimilating diploid yeast, Candida tropicalis. CtSNF1 could complement the snf1 mutant of S. cerevisiae. The previously published method for introducing the exogenous DNA into C. tropicalis was employed to construct SNF1/ snf1 heterozygote and snf1/snf1 homozygote strains. The successfully constructed SNF1/snf1 heterozygote was named KO-1. Disruption of the second CtSNF1 allele was unsuccessful, suggesting that CtSNF1 might be essential for cell viability. Therefore, in order to control the expression of CtSNF1, a strain (named KO-1G) in which the promoter region of CtSNF1 was replaced with the GAL10 promoter of C. tropicalis was constructed, and the growth of strains KO-1 and KO-1G was compared with that of the parental strain. The growth of strain KO-1 on glucose, sucrose, or acetate did not differ from the growth of the parental strain, but strain KO-1 showed a slight growth retardation on n-alkane. The growth of strain KO-1G on galactose was normal, but the cells stopped growing when transferred to glucose-, acetate-, or n-alkane-containing medium. Northern blot analysis against mRNA from the n-alkane-grown KO-1G strain demonstrated a close relationship between the presence of CtSNF1 mRNA and the growth of the cells, indicating that CtSNF1 is essential for cell viability. Moreover, mRNA levels of isocitrate lyase, which is localized in peroxisomes of C. tropicalis, were significantly affected by the level of CtSNF1 mRNA. Received: 3 May 1999 / Accepted: 14 July 1999     

Three members of the S multigene family are linked to the S locus of Brassica

Two self-incompatibility genes in Brassica, SLG and SRK (SLG encodes a glycoprotein; SRK encodes a receptor-like kinase), are included in the S multigene family. Products of members of the S multigene family have an SLG-like domain (S domain) in common, which may function as a receptor. In this study, three clustered members of the S multigene family, BcRK1, BcRL1 and BcSL1, were characterized. BcRK1 is a putative functional receptor kinase gene expressed in leaves, flower buds and stigmas, while BcRL1 and BcSL1 are considered to be pseudogenes because deletions causing frameshifts were identified in these sequences. Sequence and expression pattern of BcRK1 were most similar to those of the Arabidopsis receptor-like kinase gene ARK1, indicating that BcRK1 might have a function similar to that of ARK1, in processes such as cell expansion or plant growth. Interestingly, the region containing BcRK1, BcRL1 and BcSL1 is genetically linked to the S locus and the physical distance between SLG, SRK and the three S-related genes was estimated to be less than 610 kb. Thus the genes associated with self-incompatibility exist within a cluster of S-like genes in the genome of Brassica. Received: 15 April 1997 / Accepted: 13 June 1997     

An unstable gene controlling developmental variegation in okra (Abelmoschus esculentus (L.) Moench)

Summary Genetic studies on radiation-induced chlorina and variegated mutants of okra (Abelmoschus esculentus (L.) Moench) revealed the existence of an unstable gene. The normal green color of the leaves is controlled by duplicate genes C₁ and C₂, either of which produces the green colour. The chlorina plants are C ₁ C ₁ C ₂ C ₂. The allele c ₁ ^v is dominant to both C ₁ and C ₂ but is unstable. The homozygote c ₁ ^v c ₁ ^v c ₂ c ₂ is a normal green while the heterozygote c _i ^v c ₁ c ₂ c ₂ has a variegated phenotype as a result of the mutation of c ₁ ^v to c ₁ during development. In green plants with a c ₁ ^v c{sh1/v}c ₂ c ₂ genotype, the autonomous mutation of one of the c ₁ ^v alleles to c ₁ may take place at the pre-meiotic stage. In the variegated genotype (c ₁ ^v c ₁ c ₂ c ₂), the mutation of c ₁ to c ₁ ^v may take place in early ontogeny, thus producing green plants. The allele C ₁, when associated with c ₁ ^v in a heterozygous condition, mutates to c ₁ at the pre-meiotic stage even in the presence of the allele C ₂.     

Effects of altered α‐ and β‐branch carotenoid biosynthesis on photoprotection and whole‐plant acclimation of Arabidopsis to photo‐oxidative stress

Functions of α‐ and β‐branch carotenoids in whole‐plant acclimation to photo‐oxidative stress were studied in Arabidopsis thaliana wild‐type (wt) and carotenoid mutants, lut ein deficient (lut2, lut5), n on‐p hotochemical q uenching1 (npq1) and s uppressor of z eaxanthin‐l ess1 (szl1) npq1 double mutant. Photo‐oxidative stress was applied by exposing plants to sunflecks. The sunflecks caused reduction of chlorophyll content in all plants, but more severely in those having high α‐ to β‐branch carotenoid composition (α/β‐ratio) (lut5, szl1npq1). While this did not alter carotenoid composition in wt or lut2, which accumulates only β‐branch carotenoids, increased xanthophyll levels were found in the mutants with high α/β‐ratios (lut5, szl1npq1) or without xanthophyll‐cycle operation (npq1, szl1npq1). The PsbS protein content increased in all sunfleck plants but lut2. These changes were accompanied by no change (npq1, szl1npq1) or enhanced capacity (wt, lut5) of NPQ. Leaf mass per area increased in lut2, but decreased in wt and lut5 that showed increased NPQ. The sunflecks decelerated primary root growth in wt and npq1 having normal α/β‐ratios, but suppressed lateral root formation in lut5 and szl1npq1 having high α/β‐ratios. The results highlight the importance of proper regulation of the α‐ and β‐branch carotenoid pathways for whole‐plant acclimation, not only leaf photoprotection, under photo‐oxidative stress.     

Genetic characterization of the mei-41 locus in Drosophila melanogaster

Summary The mutagen-sensitive mutant mus(1)104 ^D1 of Drosophila melanogaster maps to a position on the X chromosome very close to the meiotic mutant mei-41 ^D5 . Both mutants have been characterized as mutagen-sensitive and defective in post-replication repair. In the present report we show by complementation studies that mus(1)104 and mus(1)103 are allelic with mei-41. In addition, two reported alleles of mus(1)104 lie between the mei-41 alleles A10 and D5. The size of the mei-41 locus is estimated to be about 0.1 centimorgans (cM). Because several alleles of mei-41 have been shown to reduce recombination and increase meiotic chromosome loss and nondisjunction, mus(1)104 ^D1 females were examined for defects in meiosis. Although there was no evidence for reduced recombination on the second chromosome in homozygous mus(1)104 ^D1 females, heterozygous mus(1)104 ^D1 /mei-41 ^>D5 and mus(1)104 ^D1 /deficiency females showed reduced levels of recombination. However, there was no evidence of an increase in nondijunction in these females.We dedicate this article to the memory of Larry Sandler, who passed away suddenly on February 7, 1987     

Cloning of caf1 +, caf2 + and caf4? + from Schizosaccharomyces pombe : their involvement in multidrug resistance, UV and pH sensitivity

We previously identified four nuclear genes (caf1 ⁺ to caf4 ⁺) in Schizosaccharomyces pombe, mutations in which can confer caffeine resistance. Here we report the cloning and sequencing of caf1 ⁺, caf2 ⁺ and caf4 ⁺. All three genes are allelic to genes (hba1 ⁺ , crm1 ⁺ and trr1 ⁺ , respectively) involved in multidrug resistance mechanisms or in stress response systems. In agreement with this the caffeine-resistant mutants caf1(hba1)-21, caf2(crm1)-3 and caf4(trr1)-83 are also resistant to brefeldin. Disruption of caf1(hba1) ⁺ and caf4(trr1) ⁺ makes cells sensitive to high pH. The overlapping ranges of pleiotropic effects and the genetic interaction detected between caf1(hba1) ⁺ and caf2(crm1) ⁺ suggest that the three genes function in interlinked systems. Received: 9 March 1998 / Accepted: 16 September 1998     

The tomato homolog of the gene encoding UV-damaged DNA binding protein 1 (DDB1) underlined as the gene that causes the<Emphasis Type="Italic"> high pigment-1</Emphasis> mutant phenotype

A tomato EST sequence, highly homologous to the human and Arabidopsis thaliana UV-damaged DNA binding protein 1 (DDB1), was mapped to the centromeric region of the tomato chromosome 2. This region was previously shown to harbor the HP-1 gene, encoding the high pigment-1 (hp-1) and the high pigment-1^w (hp-1^w) mutant phenotypes. Recent results also show that the A. thaliana DDB1 protein interacts both genetically and biochemically with the protein encoded by DEETIOLATED1, a gene carrying three tomato mutations that are in many respects isophenotypic to hp-1: high pigment-2 (hp-2), high pigment-2^j (hp-2^j) and dark green (dg). The entire coding region of the DDB1 gene was sequenced in an hp-1 mutant and its near-isogenic normal plant in the cv. Ailsa Craig background, and also in an hp-1^w mutant and its isogenic normal plant in the GT breeding line background. Sequence analysis revealed a single A⁹³¹-to-T⁹³¹ base transversion in the coding sequence of the DDB1 gene in the hp-1 mutant plants. This transversion results in the substitution of the conserved asparagine at position 311 to a tyrosine residue. In the hp-1^w mutant, on the other hand, a single G²³⁹²-to-A²³⁹² transition was observed, resulting in the substitution of the conserved glutamic acid at position 798 to a lysine residue. The single nucleotide polymorphism that differentiates hp-1 mutant and normal plants in the cv. Ailsa Craig background was used to design a pyrosequencing genotyping system. Analysis of a resource F₂ population segregating for the hp-1 mutation revealed a very strong linkage association between the DDB1 locus and the photomorphogenic response of the seedlings, measured as hypocotyl length (25<LOD score<26, R²=62.8%). These results strongly support the hypothesis that DDB1 is the gene encoding the hp-1 and hp-1^w mutant phenotypes.Communicated by R. Hagemann     

Suppression of methionine mutants in Coprinus

Summary The met-1-1 mutation in Coprinus lagopus is known to be suppressed by five recessive non-allelic suppressor genes (sup-1 to sup-5). Two of these genes complement normally in heterozygotes but the other three fail to complement each other in any combination. Four of the suppressor genes, sup-1, sup-2, sup-3 and sup-5, were tested for ability to suppress met-1-2 a second met-1 mutation. Non-identity of the two met-1 alleles was first confirmed by demonstrating intragenic recombination. The complementing suppressors, sup-1 and sup-2, proved to be allele unspecific and suppressed both met-1 mutations. The non-complementing suppressors, sup-3 and sup-5, were allele specific and could only suppress the met-1-1 mutation. This is interpreted to mean that sup-1 and sup-2 act indirectly to circumvent the metabolic lesion caused by any met-1 mutation whereas sup-3 and sup-5 are missense informational suppressors involving modified tRNA species which specifically mistranslate the met-1-1 mutant codon.     

Genetic control of chlorophyll biosynthesis: Regulation of delta-aminolevulinate synthesis in Chlamydomonas

Summary A chlorophyll-deficient mutant, br _s -1, of Chlamydomonas reinhardtii has been shown to accumulate low levels of an intermediate, protoporphyrin (PROTO), and to form light-brown colonies. A double mutant, br _s -1 r-1, accumulates 15-fold more PROTO than br _s -1 and forms dark-brown colonies. Enzymes synthesizing the first intermediate of chlorophyll, delta-aminolevulinate (ALA), from these two mutants and the wild-type are equally sensitive to inhibition by heme. The activity of ALA-synthesizing enzymes from br _s -1 r-1 is similar to that of the wild-type and is more than threefold that of br _s -1. It is proposed that the ALA-synthesizing enzymes in br _s -1 are under repression while r-1 is a mutation of the regulatory gene and consequently derepresses the synthesis of its own ALA-synthesizing enzymes. In addition, by mutagenizing br _s -1, we isolated six more double mutants having the same phenotype as br _s -1 r-1. Five of them are identical to br _s -1 r-1, the remaining one (db-10) carries a second mutation nonallelic to r-1. The ALA-synthesizing enzymes from db-10 are much less sensitive to heme inhibition than those from the wild type. It is proposed that ALA synthesis in Clamydomonas is regulated both allosterically and genetically.Abbreviations PROTO protoporphyrin - ALA delta-aminolevulinate - Mg-PROTO magnesium-protoporphyrin - GSA glutamate-1-semialdehyde     

Structural and functional characterization of <Emphasis Type="Italic">IbMYB1</Emphasis> genes in recent Japanese purple-fleshed sweetpotato cultivars

To elucidate further the genetic mechanism underlying anthocyanin accumulation in the storage roots of recent Japanese purple-fleshed sweetpotato cultivars, we compared the structure of the IbMYB1 gene in cultivar Ayamurasaki and its spontaneous mutant, AYM96, whose storage roots do not accumulate anthocyanin. Amplification of the IbMYB1 genomic fragment covering the coding sequences suggested that the genome of Ayamurasaki contained three types of IbMYB1 sequences, named IbMYB1-1, IbMYB1-2a and IbMYB1-2b, whereas AYM96 had only IbMYB1-1. Although these three IbMYB1 sequences had identical coding sequences, IbMYB1-1 had a 7-bp insertion in the first intron. IbMYB1-2a and IbMYB1-2b were characterized by a single nucleotide polymorphism in the second intron. Further cloning and sequencing of the flanking regions of these IbMYB1 sequences showed that the promoter and 3′ flanking regions of IbMYB1-2a and IbMYB1-2b were different from those of IbMYB1-1. Genetic analysis using an F₁ population derived from a cross between the purple-fleshed cultivar Murasakimasari and AYM96 suggested that IbMYB1-2 sequences are responsible for anthocyanin accumulation in the storage roots. The structural features of these three IbMYB1 sequences and identification of the IbMYB1-2null sequence, which contained sequences very similar to those of the flanking regions of IbMYB1-2a and IbMYB1-2b, but which lacked the sequence around the coding region, suggested that IbMYB1 genes in recent Japanese purple-fleshed cultivars had been established through multiple gene-duplication events.     

Polymorphism of the DQA1 promoter region (QAP) and DRB1, QAP,DQA1, DQB1 haplotypes in systemic lupus erythematosus

We have investigated the DNA polymorphism for the DQA1 promoter region (QAP) and HLA-class II DRB1, DQA1, and DQB1 genes in 178 central European patients with Systemic lupus erythematosus (SLE) using polymerase chain reaction and Dig-ddUTP labeled oligonucleotides. Increased frequencies of DRB1*02 and *03 are confirmed by DNA typing. In addition, the frequencies of DQA1*0501, *0102 and DQB1*0201, *0602 alleles are increased in the patients as compared to controls. The strongest association to SLE is found with DRB1*03 and DQB1*0201 alleles (p<10^–7, p corr. <10^–5 and p<10^–6, p corr. <10^–4, respectively). By investigating the DQA1 promoter region in the SLE patients we have detected nine different QAP variants. Increased frequencies of QAP1.2 and QAP4.1 are observed in patients as compared to controls (p <0.05, p corr. = n. s.). Analysis of linkage disquilibria demonstrates a very strong association between QAP variants and DQA1, DRB1 alleles. Certain QAP variants are completely associated with DQA1 and DRB1 alleles, whereas others can combine with different DQA1 and DRB1 alleles. All DRB1*02-positive patients and controls carry QAP1.2, and all DRB1*03-positive patients and controls carry QAP4.1. Conversely, the QAP1.2 variant appears only in DRB1*02 haplotypes, while the QAP4.1 variant can be observed in DRB1*03, *11, and *1303 haplotypes. Based on the strong linkage disequilibria between DRB1-DQA1-DQB1 genes and between DRB1-QAP-DQA1, we have deduced the four-point haplotypes for DRB1-QAP-DQA1-DQB1 in patients and controls. Two haplotypes DRB1*02-QAP1.2-DQA1*0102-DQB1*0602-and DRB1*03-QAP4.1-DQA1*0501-DQB1*0201 are significantly increased in patient as compared to controls (p<0.01, p corr. = n.s., RR = 1.8 and p <10^–7, p corr. <10^–5, RR = 3.1, respectively). The analysis of relative risks attributed to the various alleles of QAP, DQA1, and DQB1 as well as the investigation of the deduced DRB1-QAP-DQA1-DQB1 haplotypes leads to the conclusion that QAP4.1 and DQA1*0501 on the DR3 haplotypes are probably not involved in SLE susceptibility. There is no evidence for the involvement of DQ2 / dimers coded in transposition. Thus, susceptibility to SLE is on the DR3 haplotype most probably localized at DRB1 or telomeric of DRB1, while for the DR2 haplotype such orientation cannot be given. SLE study group members: M. Baur, A. Corvetta, H. Ehrfeld, J. Frey, J. R. Kalden, F. Krapf, B. Lang, G. G. Lange, K. Pirner, C. Rittner, E. Röther, P. Schneider, H. P. Seelig, S. Seuchter, W. Stangel, C. Specker, P. Späth, H. Deicher. Correspondence to: Z. Yao.     

The Arabidopsis transposable element Tag1 is widely distributed among Arabidopsis ecotypes

Tag1 is an autonomous transposable element (3.3 kb in length) first identified as an insertion in the CHL1 (NRT1) gene of Arabidopsis thaliana. Tag1 has been found in the Landsberg erecta ecotype of A. thaliana but not in Columbia or WS. In this paper, 41 additional ecotypes were examined for the presence of Tag1. Using an internal Tag1 fragment as probe, we found that DNA from 19 of the 41 ecotypes strongly hybridized to Tag1. Almost all of the Tag1-containing ecotypes had only one or two copies of Tag1 per haploid genome, as determined by Southern blot analysis. The only exception, Bf-1 from Bretagny-sur-Orge, France, had four copies. Two ecotypes, Di-G and S96, gave identical Southern blot patterns to that of Landsberg erecta and were subsequently shown to contain Tag1 at the same two positions found in Landsberg erecta (loci designated as Tag1-2 and Tag1-3). Two other ecotypes, Ag-0 and Lo-1, had a Tag1 element located at Tag1-2 but not at Tag1-3. The distance between these two loci was determined to be 0.37 cM. Analysis of DNA from two related species, A. griffithiana and A. pumila, showed that both species contain sequences that hybridize to Tag1 and that could be amplified with an oligonucleotide specific to the terminal inverted repeats of Tag1. These results show that Tag1 and related elements are present, and may be useful for insertional mutagenesis, in many A. thaliana ecotypes and several Arabidopsis species. Received: 18 August 1997 / Accepted: 9 October 1997     

Opposing effects of MYZUS PERSICAE-INDUCED LIPASE 1 and jasmonic acid influence the outcome of Arabidopsis thaliana–Fusarium graminearum interaction

Fusarium graminearum (Fg) is an important fungal pathogen of small grain cereals that can also infect Arabidopsis thaliana. In Arabidopsis, jasmonic acid (JA) signalling involving JASMONATE RESISTANT 1 (JAR1), which synthesizes JA-isoleucine, a signalling form of JA, promotes susceptibility to Fg. Here we show that Arabidopsis MYZUS PERSICAE-INDUCED LIPASE 1 (MPL1), via its influence on limiting JA accumulation, restricts Fg infection. MPL1 expression was up-regulated in response to Fg infection, and MPL1-OE plants, which overexpress MPL1, exhibited enhanced resistance against Fg. In comparison, disease severity was higher on the mpl1 mutant than the wild type. JA content was lower in MPL1-OE and higher in mpl1 than in the wild type, indicating that MPL1 limits JA accumulation. Pharmacological experiments confirmed the importance of MPL1-determined restriction of JA accumulation on curtailment of Fg infection. Methyl-JA application attenuated the MPL1-OE-conferred resistance, while the JA biosynthesis inhibitor ibuprofen enhanced resistance in mpl1. Also, the JA biosynthesis-defective opr3 mutant was epistatic to mpl1, resulting in enhanced resistance in mpl1 opr3 plants. In comparison, JAR1 was not essential for the mpl1-conferred susceptibility to Fg. Considering that methyl-JA promotes Fg growth in culture, we suggest that in part MPL1 curtails disease by limiting the availability of a plant-derived Fg growth-promoting factor.