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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 251 (1996), S. 60-68 
    ISSN: 1617-4623
    Keywords: Idiomorph ; DNA binding ; Mating type ; Bipolaris sacchari ; Cochliobolus heterostrophus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The availability of cloned genes that control sexual reproduction (mating type genes) in higher fungi has allowed us to consider the causes of failure to mate in asexual fungi. We report here that the asexual fungusBipolaris sacchari has a homolog of theMAT-2 gene of its sexual ascomycete relativeCochliobolus heterostrophus. TheB. sacchari MAT-2 sequence is highly similar to that ofC. heterostrophus MAT-2 and, in fact, functions in transgenicC. heterostrophus. Thus, the asexual nature ofB. sacchari is not due to absence or mutation ofMAT. When either of theC. heterostrophus MAT genes was transformed intoB. sacchari, the recipient could neither self nor cross with otherB. sacchari strains, in contrast to transgenicC. heterostrophus strains which can do both. Persistent asexuality ofB. sacchari, in spite of the presence of complementary functionalMAT genes, suggests that this fungus lacks genes other thanMAT which are essential for mating. Notably, the transgenicB. sacchari strains were sometimes able to initiate, but not complete, sexual development in interspecific pairings withC. heterostrophus. Transcript analysis showed that theB. sacchari MAT-2 gene is expressed in transgenicC. heterostrophus and that theC. heterostrophus MAT genes are expressed in transgenicB. sacchari. No transcript of the nativeB. sacchari MAT-2 gene was detected under any growth condition tested.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 256 (1997), S. 661-673 
    ISSN: 1617-4623
    Keywords: Key words Ascomycete ; Intron ; uORF ; UTR ; mRNA leader
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The single mating type locus (MAT ) of the heterothallic ascomycete Cochliobolus heterostrophus is composed of a pair of unlike sequences called idiomorphs, each of which encodes one MAT-specific gene (MAT-1 and MAT-2). MAT transcripts were observed in blots of poly(A)+ RNA isolated from cultures grown in minimal medium, but were not detectable after growth of the fungus in complete medium, suggesting that transcription of MAT is tightly regulated. The idiomorphs (MAT-1 = 1297-bp, MAT-2 = 1171-bp) encode transcripts of 2.2 kb (MAT-1) and 2.1 kb (MAT-2), which start 5′ and end 3′ of the idiomorph within sequences common to both mating types. Analyses of MAT-1 and MAT-2 cDNAs revealed obligatory splicing of one intron (55-bp in MAT-1, 52-bp in MAT-2) within each MAT-specific ORF and optional splicing of two introns (63 and 79-bp) in the long (approximately 0.55 kb) 5′ untranslated leader sequences; the 3′ untranslated region is 0.46 kb long. Transcription start sites were found 5′ of, and within, the 79-bp intron. Optional splicing of the upstream introns and at least two transcription start sites result in three types of transcript: Type I with both 5′ introns spliced, Type II with only the 63-bp intron spliced, and Type III with neither 5′ intron spliced. The three transcript types are distinguished by various combinations of four short ORFs encoded by the corresponding genomic DNA, in the leader sequences of the MAT mRNAs. The transcript structure suggests several mechanisms by which expression of the MAT genes might be regulated at the level of translation during sexual development.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1617-4623
    Keywords: Key words HMG box ; Transcriptional regulator ; 3′ UTR ; DNA-binding protein ; Heterothallic ascomycete
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To determine the number of proteins required for mating type (MAT) locus-regulated control of mating in Cochliobolus heterostrophus, MAT fragments of various sizes were expressed in MAT deletion strains. As little as 1.5 kb of MAT sequence, encoding a single unique protein in each mating type (MAT-1 and MAT-2), conferred mating ability, although an additional 160 bp of 3′ UTR was needed for production of ascospores. No other mating type-specific genes involved in mating identity or fertility were found. Thus, although homologs of the C. heterostrophus MAT-1 and MAT-2 genes exist in the filamentous ascomycetes Neurospora crassa and Podospora anserina, C. heterostrophus does not appear to have mating type-specific homologs of two additional genes required by both N. crassa and P.␣anserina for successful sexual reproduction. Three genes were identified in the common DNA flanking the MAT locus: a gene encoding a GTPase-activating protein and an ORF of unknown function lie 5′ while a β-glucosidase encoding gene lies found 3′. None of these genes appears to be involving in the mating process.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Glycine (Rhizobium infection) ; Infection thread ; Rhizobium ; Root hair infection ; Symbiosis (legume-Rhizobium)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The location and topography of infection sites in soybean (Glycine max (L.) Merr.) root hairs spot-inoculated with Rhizobium japonicum have been studied at the ultrastructural level. Infections commonly developed at sites created when the induced deformation of an emerging root hair caused a portion of the root-hair cell wall to press against an adjacent epidermal cell, entrapping rhizobia within the pocket between the two host cells. Infections were initiated by bacteria which became embedded in the mucigel in the enclosed groove. Infection-thread formation in soybean appears to involve degradation of mucigel material and localized disruption of the outer layer of the folded hair cell wall by one or more entrapped rhizobia. Rhizobia at the site of penetration are separated from the host cytoplasm by the host plasmalemma and by a layer of wall material that appears similar or identical to the normal inner layer of the hair cell wall. Proliferation of the bacteria results in an irregular, wall-bound sac near the site of penetration. Tubular infection threads, bounded by wall material of the same appearance as that surrounding the sac, emerge from the sac to carry rhizobia roughly single-file into the hair cell. Growing regions of the infection sac or thread are surrounded by host cytoplasm with high concentrations of organelles associated with synthesis and deposition of membrane and cell-wall material. The threads follow a highly irregular path toward the base of the hair cell. Threads commonly run along the base of the hair cell for some distance, and may branch and penetrate into subjacent cortical cells at several points in a manner analagous to the initial penetration of the root hair.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 115 (1983), S. 122-128 
    ISSN: 1615-6102
    Keywords: Inoculation ; Nodulation ; Rhizobium japonicum ; Soybean ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A method has been developed to inoculate soybean roots in the zone of susceptible root cells with nanoliter droplets containing the symbiont,Rhizobium japonicum. The position of inoculation is marked by means of ion exchange beads which adhere to the root surface. InoculatedR. japonicum cells from a nanoliter droplet spread over an area of approximately 35 host epidermal cells and have a high probability (∼ 80%) of inducing nodule formation at exactly the point of inoculation. The spot inoculation method is highly efficient in terms of the number of bacteria required to achieve a given probability of nodule formation. The method greatly facilitates ultrastructural analysis of the infection process and is useful for certain physiological studies of infection.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 251 (1996), S. 60-68 
    ISSN: 1617-4623
    Keywords: Key words Idiomorph ; DNA binding ; Mating type ; Bipolaris sacchari ; Cochliobolus heterostrophus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The availability of cloned genes that control sexual reproducti on (mating type genes) in higher fungi has allowed us to consider the causes of failure to mate in asexual fungi. We report here that the asexual fungus Bipolaris sacchari has a homolog of the MAT-2 gene of its sexual ascomycete relative Cochliobolus heterostrophus. The B. sacchari MAT-2 sequence is highly similar to that of C. heterostrophus MAT-2 and, in fact, functions in transgenic C. heterostrophus. Thus, the asexual nature of B. sacchari is not due to absence or mutation of MAT. When either of the C. heterostrophus MAT genes was transformed into B. sacchari, the recipient could neither self nor cross with other B. sacchari strains, in contrast to transgenic C. heterostrophus strains which can do both. Persistent asexuality of B. sacchari, in spite of the presence of complementary functional MAT genes, suggests that this fungus lacks genes other than MAT which are essential for mating. Notably, the transgenic B. sacchari strains were sometimes able to initiate, but not complete, sexual development in interspecific pairings with C. heterostrophus. Transcript analysis showed that the B. sacchari MAT-2 gene is expressed in transgenic C. heterostrophus and that the C. heterostrophus MAT genes are expressed in transgenic B. sacchari. No transcript of the native B. sacchari MAT-2 gene was detected under any growth condition tested.
    Type of Medium: Electronic Resource
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