No abstract

No abstract

The complete nucleotide sequence of RNA beta from the type strain of barley stripe mosaic virus (BSMV) has been determined. The sequence is 3289 nucleotides in length and contains four open reading frames (ORFs) which code for proteins of Mr 22,147 (ORF1), Mr 58,098 (ORF2), Mr 17,378 (ORF3), and Mr 14,119 (ORF4). The predicted N-terminal amino acid sequence of the polypeptide encoded by the ORF nearest the 5'-end of the RNA (ORF1) is identical (after the initiator methionine) to the published N-terminal amino acid sequence of BSMV coat protein for 29 of the first 30 amino acids. ORF2 occupies the central portion of the coding region of RNA beta and ORF3 is located at the 3'-end. The ORF4 sequence overlaps the 3'-region of ORF2 and the 5'-region of ORF3 and differs in codon usage from the other three RNA beta ORFs. The coding region of RNA beta is followed by a poly(A) tract and a 238 nucleotide tRNA-like structure which are common to all three BSMV genomic RNAs.

Spinosyns, a novel class of insect active macrolides produced by Saccharopolyspora spinosa, are used for insect control in a number of commercial crops. Recently, a new class of spinosyns was discovered from S. pogona NRRL 30141. The butenyl-spinosyns, also called pogonins, are very similar to spinosyns, differing in the length of the side chain at C-21 and in the variety of novel minor factors. The butenyl-spinosyn biosynthetic genes (bus) were cloned on four cosmids covering a contiguous 110-kb region of the NRRL 30141 chromosome. Their function in butenyl-spinosyn biosynthesis was confirmed by a loss-of-function deletion, and subsequent complementation by cloned genes. The coding sequences of the butenyl-spinosyn biosynthetic genes and the spinosyn biosynthetic genes from S. spinosa were highly conserved. In particular, the PKS-coding genes from S. spinosa and S. pogona have 91-94% nucleic acid identity, with one notable exception. The butenyl-spinosyn gene sequence codes for one additional PKS module, which is responsible for the additional two carbons in the C-21 tail. The DNA sequence of spinosyn genes in this region suggested that the S. spinosa spnA gene could have been the result of an in-frame deletion of the S. pogona busA gene. Therefore, the butenyl-spinosyn genes represent the putative parental gene structure that was naturally engineered by deletion to create the spinosyn genes.

SUMMARY Quinoxyfen is a protectant fungicide which controls powdery mildew diseases by interfering with germination and/or appressorium formation. Mutants of barley powdery mildew, Blumeria graminis f.sp. hordei, which are resistant to quinoxyfen produce fewer conidia, which germinate and form appressoria more promiscuously than do the prolific numbers of wild‐type spores. This suggests that resistance bypasses host recognition signals. RT‐PCR profiles of signal transduction genes, recorded during wild‐type germling morphogenesis, reveals that quinoxyfen alters the accumulation of Protein Kinase C (pkc), pkc‐like and catalytic subunit of Protein Kinase A (cpka) transcripts. Differential display‐reverse transcription PCR identified a gene transcript in wild‐type conidia that was absent, or much less abundant, in conidia from quinoxyfen‐resistant mutants. This mRNA was not detectable 24 h after wild‐type conidia were inoculated on to barley. It encodes a GTPase activating protein (GAP), which may interact with a small molecular weight Ras‐type GTP binding protein. In the presence of quinoxyfen, the gap mRNA remains throughout germling morphogenesis. The involvement of GAP in resistance suggests that quinoxyfen inhibits mildew infection by disrupting early cell signalling events.