Cloning of DNA fragments specific to the pathotype and race of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Japanese isolates of Verticillium dahliae, a causal agent of wilt disease in many plants, are classifiable into pathotypes based on their pathogenicity. Because these pathotypes are morphologically indistinguishable, establishing a rapid identification method is very important for the control of this pathogen in Japan. For cloning DNA fragments that are useful for identification and specific detection of V. dahliae pathotypes, we performed random amplified polymorphic DNA (RAPD) analyses using various isolates. One polymerase chain reaction (PCR) product, E10-U48, was specific to isolates pathogenic to sweet pepper. The other product, B68-TV, was specific to race 1 of isolates pathogenic to tomato. The specificity of these sequences was confirmed by genomic Southern hybridization. Further analyses revealed that the region peripheral to B68-TV obtained from the genomic DNA library includes the sequence specific to all isolates pathogenic to tomato (races 1 and 2). Moreover, sequence tagged site (STS) primers designed from B68-TV and its peripheral region showed race-specific and pathotype-specific amplification in a PCR assay. The probes and primers obtained in this study are likely to be useful tools for the identification and specific detection of pathotypes and races of V. dahliae. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB095266.     

Regulation of stony hard peach softening with ACC treatment

Controlling the rate of fruit softening in melting-flesh peaches is a primary goal of the fruit industry. Stony hard (SH) peach varieties lack the ability to synthesize 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, which is required for fruit maturation. SH peaches thus have crisp flesh that remains firm during ripening. In this study, we developed a simple technique to stimulate fruit softening by a single spray application of ACC at a concentration of 10–20 mM, which was sufficient to allow ethylene synthesis and fruit softening. Higher concentrations of ACC increased ethylene production, and made the fruit softer. Ethylene synthesis was limited to the first 2–3 d after ACC treatment, after which fruit ceased softening and retained its remaining firmness. These results indicate that a single application of ACC solution can be used to regulate the process of fruit softening in SH peaches.     

The extinction of genetic resources of Asian wild rice, Oryza rufipogon Griff.: A case study in Thailand

Natural populations of wild rice, Oryza rufipogon Griff., are now threatened with the disturbance of their natural habitats by various human activities. To obtain basic information on genetic erosion or loss of genetic diversity in wild rice, we investigated how environmental changes of habitat affected the genetic structure of its natural population at a study site in the central plain of Thailand. During 10 years from 1985 to 1994, the wild-rice population at this site was seriously destroyed and fragmented. Using two sets of seed sample collected in 1985 and 1994 from the same population, allozyme variability at 17 loci of 11 enzymes were examined. Isozyme genotypes of mother plants of seed samples were estimated by the segregation in each progeny, and we calculated genetic parameters for the population. Gene diversity severely decreased in the 1994 sample compared with the 1985 sample. It is supposed that declining and fragmentation of the wild rice population, which happened during the 10 years, caused loss of genetic variability and forced the habitually outbreeding plants to inbreed, accelerating a reduction in gene variability. Pgi1-1 allele which was common in Indica rice cultivars of this region was found in the wild rice plants growing at the side of rice fields. Probably, introgression has occurred between wild and cultivated rice plants, and consequently the intrinsic nature of wild rice was gradually blurred by cultivar genes. We must realize that the genetic erosion of wild rice is rapidly proceeding and that an action for their conservation in natural environment, so called in situ conservation, is urgently needed.     

Phylogenetic differentiation of cultivated rice,XXIII. Potentiality of wild progenitors to evolve the Indica and Japonica types of rice cultivars

Summary Studies of intermediate wild-cultivated plants suggested that the Indica-Japonica differentiation of cultivars of Oryza sativa L. has taken place with the domestication of wild plants. The wild progenitor, which is considered to be the Asian form of O. perennis Moench, shows no trace of differentiation into two such types as the Indica and Japonica. To elucidate the potentiality of the wild progenitor to evolve the two types when domesticated, the selfed progenies from an Indica × wild and a japonica × wild cross were examined with regard to three characters known to be useful for distinguishing between the two types. The data showed that Japonica-like plants were derived from the Indica × wild cross and Indica-like plants from the Japonica × wild cross. The Indica-Japonica differentiation was considered to have resulted from selection of incipient domesticates having such potentiality under different climatic conditions; the dynamics was discussed.     

Measurement of ammonia volatilization loss using a dynamic chamber technique: A case study of surface-incorporated manure and ammonium sulfate in an upland field of light-colored Andosol

The present study aimed to elucidate ammonia (NH₃) volatilization loss following surface incorporation (0–15 cm mixing depth) of nitrogen (N) fertilizer in an upland field of light-colored Andosol in central Japan. A dynamic chamber technique was used to measure the NH₃ effluxes. Poultry manure, pelleted poultry manure, cattle manure, pelleted cattle manure and ammonium sulfate were used as N fertilizers for basal fertilization to a bare soil with surface incorporation. All three experiments in summer and autumn 2007 and in summer 2008 showed negligible NH₃ volatilization losses following the application of all N fertilizers with the same application rate of 120 kg N ha⁻¹ as total N; these negligible losses were primarily ascribed to chemical properties of the soil, that is, its high cation exchange capacity (283 mmol_c kg⁻¹ dry soil) and relatively low pH(H₂O) (5.9). In addition, the surface incorporation, the very small ratio of ammoniacal N to total N for the manure, and the decrease in soil pH to ≤5.5 following applications of ammonium sulfate were also advantageous to the inhibition of NH₃ volatilization loss from the field-applied N fertilizers.     

Divergent evolution of wild and cultivated subspecies of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> as revealed by the study of <Emphasis Type="Italic">PolA1</Emphasis> gene

Triticum timopheevii (genome symbol AAGG) comprises two subspecies, cultivated ssp. timopheevii, and wild ssp. armeniacum. These two subspecies are considered as allotetraploids of AA genome from Triticum diploid species and SS genome from Aegilops species. The difference in genome symbol (G vs. S) is due to wide genetic variations among four SS genome species, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. speltoides. In order to study the origin of T. timopheevii, we compared 19th intron (PI19) sequence of the PolA1 gene, encoding the largest subunit of RNA polymerase I. Two different sized DNA fragments containing PI19 sequences (PI19A and PI19G) were amplified both in ssp. timopheevii and ssp. armeniacum. Shorter PI19A (112 bp) sequences of both subspecies were identical to PI19 sequences of two AA species, T. monococcum and T. urartu. Interestingly, the longer PI19G (241–243 bp) sequences of ssp. armeniacum showed more similarity to PI19 sequences of Ae. speltoides whereas ssp. timopheevii showed more similarity to PI19 sequences of other three SS genome species. The results indicated that two subspecies of T. timopheevii, ssp. armeniacum or ssp. timopheevii, might have arisen independently by allotetraploidization of AA genome with Ae. speltoides or one of the remaining three Aegilops species, respectively.     

Negative feedback regulation ensures the one receptor-one olfactory neuron rule in mouse

In the mouse olfactory system, each olfactory sensory neuron (OSN) expresses only one odorant receptor (OR) gene in a monoallelic and mutually exclusive manner. Such expression forms the genetic basis for OR-instructed axonal projection of OSNs to the olfactory bulb of the brain during development. Here, we identify an upstream cis-acting DNA region that activates the OR gene cluster in mouse and allows the expression of only one OR gene within the cluster. Deletion of the coding region of the expressed OR gene or a naturally occurring frame-shift mutation allows a second OR gene to be expressed. We propose that stochastic activation of only one OR gene within the cluster and negative feedback regulation by that OR gene product are necessary to ensure the one receptor-one neuron rule.     

亚洲栽培稻两大亚种之间同工酶基因型的主要区别

选择亚洲重要稻作区的中国、日本、朝鲜、印度、泰国、缅甸、尼泊尔等地的栽培稻品种89份及野生稻与栽培稻的中间稻系14份为材料,根据形态进行分类并检测其同工酶。从结果看,Amp-2、Acp-1和Cat-1三个同工酶对于籼粳分类有重要的作用。