RESUMO
A rice gene, OsAPT2, which encodes a putative adenine phosphoribosyl transferase (APRT), was cloned and characterized. Analysis of the cDNA and genomic sequences revealed seven exons and six introns in the OsAPT2. The deduced amino acid sequence of OsAPT2 is highly homologous to those of previously isolated APRTs. RT-PCR analysis indicated that the OsAPT2 transcript in the young panicles of 'Annong S-1' is down-regulated at 29 degrees C, the critical temperature for induction of 'Annong S-1' fertility conversion. Since the panicle is likely the thermo-sensitive organ at the early stages of pollen fertility alternation, the observed heat-induced change in the OsAPT2 expression pattern in young panicles may mediate, at least in part, thermo-sensitive genic male sterility (TGMS) in 'Annong S-1'. An antisense strategy was used to suppress the expression of the OsAPT2 homolog in Arabidopsis, and the obtained homozygous transgenic plants contained lower AMP content, displayed lower pollen germination rates and exhibited some abnormalities in leaf phenotypes and flowering timing. These data suggest that OsAPT2 is likely to be involved in TGMS in the rice line 'Annong S-1'.
Assuntos
Adenina Fosforribosiltransferase/genética , Oryza/genética , Adenina/química , Adenina Fosforribosiltransferase/química , Adenosina/química , Monofosfato de Adenosina/metabolismo , Alelos , Sequência de Aminoácidos , Northern Blotting , Clonagem Molecular , Biologia Computacional/métodos , Primers do DNA/química , DNA Complementar/metabolismo , Regulação para Baixo , Éxons , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Vetores Genéticos , Homozigoto , Temperatura Alta , Íntrons , Luz , Modelos Genéticos , Dados de Sequência Molecular , Nucleotídeos/química , Oligonucleotídeos Antissenso/química , Oryza/enzimologia , Fenótipo , Folhas de Planta/metabolismo , Fenômenos Fisiológicos Vegetais , Pólen/metabolismo , Reação em Cadeia da Polimerase , Estrutura Secundária de Proteína , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Temperatura , TransgenesRESUMO
Cloning of plant disease resistant genes is greatly helpful for disease resistant breeding in plants and the insight of resistance mechanism. However, there are less relevant researches in peach [prunus persica (L.) Batch]. In this study, four NBS-LRR type resistance gene analogs (RGAs) were cloned from genomic DNA of peach. The PNBS2 fragment was also amplified from peach cDNA and the full-length cDNA of PNBS2 (PRPM1, GenBank accession no. AY599223) has been cloned. Sequence analysis indicated that the cDNA of PRPM1 is 3007 bp in length and that the contained ORF encodes for a polypeptide of 917 amino acids. Compared with known NBS-LRR genes, it presented relatively high amino acid sequence identity. The polypeptide has typical structure of non-TIR-NBS-LRR genes, with NB-ARC, LZ, LRR and transmembrane domains. Southern analysis indicated that the PRPM1 gene might be a single copy in peach genome. Northern blot and RT-PCR analysis showed that the expression of PRPM1 was not induced by salicylic acid (SA) in peach young leaves. The isolation of putative resistance genes from peach provided useful bases for studying the structure and function of peach disease-resistance relating genes and disease resistant genetic breeding in peach.
Assuntos
Imunidade Inata , Doenças das Plantas/genética , Proteínas/genética , Prunus/genética , Sequência de Aminoácidos , Sequência de Bases , Sítios de Ligação , Northern Blotting , Southern Blotting , Mapeamento Cromossômico , Clonagem Molecular , DNA/química , DNA Complementar/metabolismo , Biblioteca Gênica , Proteínas de Repetições Ricas em Leucina , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/química , Reação em Cadeia da Polimerase , Estrutura Terciária de Proteína , RNA/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ácido Salicílico/metabolismo , Homologia de Sequência de Aminoácidos , Fatores de TempoRESUMO
beta-1,3-glucanase(BG2) is one of the pathogensis-related-proteins(PR). Study of these proteins and their related genes is one of the hot points in plant genetic engineering of disease resistance for a long time. In this research, specific primers were designed with the enzyme cleavage site of Spe I in its forward one and Not I site in the backward according to the BG2 gene sequence. Using this pair of primers, BG2 gene, which was contained in the plasmid of pRTL2, was amplified and confirmed by sequencing the amplified fragment inserted into T-easy vector. The positive clone containing BG2 gene was digested with the enzymes of Spe I/Not I and then BG2 gene was inserted into the Xba I/Not I sites of super expression binary vector pATC940. The reconstructed expression vector named as pATCBG2 was introduced into the wheat of Longfumai10 and Longfumai3 (Triticum aestivum L. em. Thell) through the particle gun transformation method. The Kanamysin resistant (Km') transformants were obtained. PCR, Dot-blotting and PCR-Southern hybridization analysis showed that the BG2 gene was integrated into the genome of wheat. Result of pathogen inoculation assay on the transgenic plants showed that the transgenic plants had a higher resistant disease score of 1-2 grade than the control.
Assuntos
Vetores Genéticos/genética , Glucana 1,3-beta-Glucosidase/genética , Triticum/genética , Ascomicetos/crescimento & desenvolvimento , Southern Blotting , Clonagem Molecular/métodos , DNA de Plantas/genética , DNA de Plantas/metabolismo , Desoxirribonuclease EcoRI/metabolismo , Desoxirribonuclease HindIII/metabolismo , Imunidade Inata/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Plantas Geneticamente Modificadas , Transformação Genética , Triticum/microbiologiaRESUMO
Southern corn rust (SCR) is a destructive disease in maize. The inbred line Qi319 is highly resistant to southern corn rust. SSR technique was employed to preliminary mapping of the resistance gene. Bulked segregant analysis revealed that two primers, phi 118 and phi 041, amplified polymorphic bands. SSR analysis on populations indicated the two primers were linked to the rust resistance gene, which was mapped on the short arm of chromosome 10. In addition, comparative analysis of the amplification bands among different populations revealed that the amplification products with the same primer in different populations were dissimilar. This result indicates that the genetic background may affect results of gene mapping and tagging. So, it is important to select suitable population to performing molecular marker analysis and gene mapping.
Assuntos
Basidiomycota/crescimento & desenvolvimento , Mapeamento Cromossômico/métodos , Zea mays/genética , Cromossomos de Plantas/genética , DNA de Plantas/genética , Imunidade Inata/genética , Repetições de Microssatélites/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Polimorfismo de Fragmento de Restrição , Zea mays/microbiologiaRESUMO
The study of plant male sterility plays an important role on utilization of heterosis. This paper reviews the current status of the studies of the heredity and mapping of the male sterile genes in wheat and the gene engineering of wheat male sterility. The application of male sterility in wheat breeding is discussed.
