A premature senescence
and death 128 (psd128) mutant was isolated from an ethyl methane sulfonate‐induced rice IR64 mutant bank. The premature senescence phenotype appeared at the six‐leaf stage and the plant died at the early heading stage. psd128 exhibited impaired chloroplast development with significantly reduced photosynthetic ability, chlorophyll and carotenoid contents, root vigor, soluble protein content and increased malonaldehyde content. Furthermore, the expression of senescence‐related genes was significantly altered in psd128. The mutant trait was controlled by a single recessive nuclear gene. Using map‐based strategy, the mutation Oryza sativa cell division cycle 48 (OsCDC48) was isolated and predicted to encode a putative AAA‐type ATPase with 809 amino‐acid residuals. A single base substitution at position C2347T in psd128 resulted in a premature stop codon. Functional complementation could rescue the mutant phenotype. In addition, RNA interference resulted in the premature senescence and death phenotype. OsCDC48 was expressed constitutively in the root, stem, leaf and panicle. Subcellular analysis indicated that OsCDC48:YFP fusion proteins were located both in the cytoplasm and nucleus. OsCDC48 was highly conserved with more than 90% identity in the protein levels among plant species. Our results indicated that the impaired function of OsCDC48 was responsible for the premature senescence and death phenotype.
A stable inherited rice spotted-leaf mutant HM47 derived from an EMS-induced IR64 mutant bank was identified. The mutant expressed hypersensitive response (HR)-like symptoms throughout its whole life from the first leaf to the flag leaf, without pathogen invasion. Initiation of the lesions was induced by light under natural summer field conditions. Expression of pathogenesis-related genes including PAL, PO-C1, POX22.3 and PBZ1 was enhanced significantly in association with cell death and accumulation of H 2 O 2 at and around the site of lesions in the mutant in contrast to that in the wild-type (WT). Disease reaction to Xanthomonas oryzae pv. oryzae from the Philippines and China showed that HM47 is a broad-spectrum disease-resistant mutant with enhanced resistance to multiple races of bacterial blight pathogens tested. An F 2 progeny test showed that bacterial blight resistance to race HB-17 was cosegregated with the expression of lesions. Genetic analysis indicated that the spotted-leaf trait was controlled by a single recessive gene, tentatively named
A rice spotted-leaf mutant was isolated from an ethane methyl sulfonate (EMS) -induced IR64 mutant bank. The mutant, designated as spl30 (spotted-leaf30), displayed normal green leaf color under shade but exhibited red-brown lesions under natural summer field conditions. Initiation of the lesions was induced by light and the symptom was enhanced at 33 (°) C relative to 26 (°) C. Histochemical staining did not show cell death around the red-brown lesions. Chlorophyll contents in the mutant were significantly lower than those of the wild type while the ratio of chlorophyll a/b remained the same, indicating that spl30 was impaired in biosynthesis or degradation of chlorophyll. Disease reaction patterns of the mutant to Xanthomonas oryzae pv. oryzae were largely unchanged to most races tested except for a few strains. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively named spl30(t), which co-segregated with RM15380 on chromosome 3, and was delimited to a 94 kb region between RM15380 and RM15383. Spl30(t) is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region. The genetic data and recombination populations provided in this study will enable further fine-mapping and cloning of the gene.
To understand the development of rice leaf blades, we identified a new rolled-leaf mutant, w32, from indica cultivar IR64 through EMS mutagenesis. The mutant showed a stable rolled-leaf phenotype throughout the life cycle. Two F2 populations were developed by crossing w32 to cultivar IR24 and PA64. Genetic analysis showed that the rolled-leaf phenotype was controlled by a single recessive gene. To determine the location of the gene, bulked segregant analysis was carried out using mutant and wild-type DNA pools and 1846 mutant-type F2 individuals derived from the cross w32/PA64 were genotyped to locate the gene on the short arm of chromosome 7. The rolled-leaf gene, tentatively named rl11(t), is likely a new gene as no other rolled-leaf genes have been identified near the region. By developing new SSR and InDel markers, the gene was delimited to a 52 kb region near the end of the short chromosome arm. Further fine mapping and cloning of the gene are currently underway.
A spotted-leaf mutant of rice HM143 was isolated from an EMS-induced IR64 mutant bank. Brown lesions randomly distributed on leaf blades were observed about 3 wk after sowing. The symptom lasted for the whole plant growth duration. Histochemical analysis indicated that cell death occurred in and around the site of necrotic lesions accompanied with accumulation of hydrogen hyperoxide. Agronomic traits were largely similar to the wild type IR64 except seed setting rate and 1000-grain weight which were significantly decreased in the mutant. Disease resistance of the mutant to multiple races of Xanthomonas oryzae pv. oryzae was significantly enhanced. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively termed spl HM143. In addition, using molecular markers and 1023 mutant type individuals from an F2 segregating population derived from the cross HM143/R9308, the spotted-leaf gene was finally delimited to an interval of 149 kb between markers XX25 and ID40 on the long arm of chromosome 4. Spl HM143 is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region.
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