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101.
Masahiro Shishido Itsuki Kubota Tatsuya Ohashi Toshiyuki Usami 《Journal of General Plant Pathology》2013,79(1):18-27
We developed a real-time PCR assay using a TaqMan probe (TM-qPCR) for specific detection and quantification of Phomopsis sclerotioides, causal agent of black root rot of cucurbit crops. The design of the primer sets and hybridization probe was based on the internal transcribed spacer region of the ribosomal DNA. The TM-qPCR assay was compared with a conventional, standard PCR (sPCR) assay and on a quantitative real-time PCR (SG-qPCR) assay based on SYBR Green I. The TM-qPCR assay had a detection limit of ca. 0.4 fg of P. sclerotioides DNA, which was approximately 100 times more sensitive than the sPCR assay and almost equivalent to the SG-qPCR assay. The TM-qPCR and SG-qPCR assays both were able to detect various quantities of P. sclerotioides DNA from diseased plants and infested soils, including DNA levels that were not detectable by the sPCR assay. However, the TM-qPCR was advantageous for samples containing PCR-inhibiting substances because its multiplex real-time PCR function allows the adjustment of cycle threshold values with an internal control. Based on the high specificity and sensitivity required for analyzing DNA in natural samples, the newly developed TM-qPCR assay was the most reliable tool for rapidly detecting and quantifying P. sclerotioides in plant and soil samples. 相似文献
102.
Masahiro Hosono Shigeki Sugawara Takeo Tatsuta Toshiyuki Hikita Junko Kominami Sachiko Nakamura-Tsuruta Jun Hirabayashi Sarkar M. A. Kawsar Yasuhiro Ozeki Sen-itiroh Hakomori Kazuo Nitta 《Fish physiology and biochemistry》2013,39(6):1619-1630
Osmerus (Spirinchus) lanceolatus egg lectin (OLL) is a member of the rhamnose-binding lectin (RBL) family which is mainly found in aqueous beings. cDNA of OLL was cloned, and its genomic architecture was revealed. The deduced amino acid (aa) sequence indicated that OLL was composed of 213 aa including 95 aa of domain N and 97 aa of domain C. N and C showed 73 % sequence identity and contained both -ANYGR- and -DPC-KYL-peptide motifs which are conserved in most of the RBL carbohydrate recognition domains. The calculated molecular mass of mature OLL was 20,852, consistent with the result, and 20,677.716, from mass spectrometry. OLL was encoded by eight exons: exons 1 and 2 for a signal peptide; exons 3–5 and 6–8 for N- and C-domains, respectively. Surface plasmon resonance spectrometric analyses revealed that OLL showed comparable affinity for Galα- and β-linkages, whereas Silurus asotus lectin (SAL), a catfish RBL, bound preferentially to α-linkages of neoglycoproteins. The Kd values of OLL and SAL against globotriaosylceramide (Gb3) were 1.69 × 10?5 M for and 2.81 × 10?6 M, respectively. Thus, the carbohydrate recognition property of OLL is slightly different from that of SAL. On the other hand, frontal affinity chromatography revealed that both OLL and SAL interacted with only glycolipid-type oligosaccharides such as Gb3 trisaccharides, not with N-linked oligosaccharides. The domain composition of these RBLs and an analytical environment such as the “cluster effect” of a ligand might influence the binding between RBL and sugar chains. 相似文献
103.
Guang-Xi Wang Toshiyuki Imaizumi Wei Li Hiromasa Saitoh Ryohei Terauchi Takanori Ohsako Tohru Tominaga 《Pesticide biochemistry and physiology》2008,92(1):24-29
TILLING (Targeting Induced Local Lesions IN Genomes) is a low-cost, high-throughput reverse genetic technique that employs a mismatch-specific endonuclease CEL-1 to discover induced point mutations in the genes of interest. The use of the TILLING technique to survey natural variation in genes is called EcoTILLING. Here, we report a modified EcoTILLING method for the discovery of mutations in multigene family, which we coin “Self-EcoTILLING” by using an allotetraploid Monochoria vaginalis ALS multigene family as an example. The mutations could be detected by TILLING of PCR products resulting from the primers specific to both Als1 and Als3 without involving the experimental step of mixture of reference and query DNA. Either of the two co-amplified loci could serve as reference DNA to the other. We demonstrate with this example that Self-EcoTILLING is a fast, reliable and economical technique of detecting single-nucleotide mutations in polyploid plants containing multigene family. 相似文献
104.
The chromosome number and electrophoretic karyotype of Japanese isolates of Verticillium dahliae were investigated. In a genomic Southern blot analysis of seven isolates probed with a telomere consensus sequence (TTAGGG)5, 12 or 14 bands were observed. Furthermore, pulsed-field gel electrophoresis (PFGE) of these isolates revealed five or six
chromosomal bands. A band (approx. 3.5 Mbp) common to all isolates apparently contained more than two chromosomes. From these
results, we concluded that each isolate’s chromosome number is six (an eggplant pathotype isolate) or seven (all isolates
of tomato and sweet pepper pathotypes). Although the chromosome sizes differed among isolates, karyotypes were similar within
tomato and sweet pepper pathotypes. A small chromosome (approx. 1.8 Mbp) was observed only in the sweet pepper pathotype.
Subsequent PFGE-Southern hybridization analyses revealed that the three DNA fragments specific to tomato pathotype are located
on the same chromosome. These results suggest that the tomato-pathotype-specific DNA sequences might coexist on one chromosome. 相似文献
105.
Toshiyuki IBARAKI Naohiko KUROYANAGI Masaharu MURAKAMI 《Soil Science and Plant Nutrition》2009,55(3):421-427
Some indica rice varieties are potential phytoextractors for paddy fields polluted with Cd because of their high biomass and because they can accumulate Cd to moderate levels in their shoots. To establish a practical phytoextraction system, phytoextraction using two indica rice cultivars (MORETSU and IR-8) was carried out in a paddy field polluted with moderate Cd levels (2.91 and 2.52 mg kg−1 , respectively). The Cd concentration and Cd uptake of MORETSU increased when irrigation water was drained at the maximum tillering stage, and the paddy soil was under oxidative conditions until harvesting. The Cd uptake of MORETSU and IR-8 increased and reached 516 and 657 g ha−1 , respectively, at the beginning of October. After phytoextraction using these high Cd accumulating rice varieties for 2 years, the Cd concentration in the paddy field decreased by 18% compared with the initial Cd concentration. The Cd concentration in the rice grains of a japonica ordinary rice variety (HINOHIKARI) subsequently grown on the field after the phytoextraction was lower than the concentration in rice grown on a non-phytoextracted field. These results suggest that phytoextraction using high Cd accumulating rice varieties with early drainage of irrigation water is a practical remediation system for moderate Cd polluted paddy fields in southwest Japan. 相似文献
106.
Susumu S. ABE Sadahiro YAMAMOTO Toshiyuki WAKATSUKI 《Soil Science and Plant Nutrition》2009,55(4):514-522
Termites play a significant role in soil-forming processes of the tropics. The influence of termites on pedogenesis as affected by the toposequence, however, has rarely been explored. We investigated the soil physicochemical and morphological characteristics of epigeal mounds constructed by Macrotermes bellicosus (Smethman) compared with those of surrounding pedons along a toposequence (bottom, fringe and upland sites) of an inland valley in central Nigeria. The physicochemical and morphological properties of the mound soils varied according to structural units but were generally different from those of the adjacent pedons. The differences included finer texture, higher electrical conductivity, total N, exchangeable bases (Ca, Mg and K) and effective cation exchange capacity and lower C/N ratio and exchange acidity in the mound than the pedon at each toposequence position. This tendency to modify the soil properties was more prominent in the nest body where the termites actually live, that is, in the hives, royal cell and base-plate, than in the soils below the nest and the other mound parts, that is, the external wall, internal wall and pillars. We found this trend to a greater or lesser degree at all toposequence positions. Our findings suggest that: (1) M. bellicosus can manipulate the mound soils according to functional applications of structure units or environmental requirements for its livelihood, regardless of local soils; (2) M. bellicosus makes ecological patches (hot spots) at all toposequence positions in the same measure; (3) the influence of M. bellicosus on the pedogenesis is reduced in the lowlands compared with the uplands because the number and volume of the mounds were substantially lower in the bottom and fringe sites compared with the upland site. 相似文献
107.
108.
Toru Kondo Mamoru Satou Toshiyuki Morikawa Yoshiaki Chikuo 《Journal of General Plant Pathology》2014,80(1):99-102
Brown root rot of Russell prairie gentian was observed in the Aomori Prefecture, Japan in April 2011. The fungal isolate from the diseased root was identified as Subplenodomus drobnjacensis on the basis of its morphological characteristics and nucleotide sequences. The isolate induced similar root rot symptoms when inoculated in healthy Russell prairie gentian plants. We proposed the name “brown root rot” for this disease. 相似文献
109.
Mitsuhiro Aihara Syun-ichi Urayama Minh Tuong Le Yu Katoh Tomoya Higashiura Toshiyuki Fukuhara Tsutomu Arie Tohru Teraoka Ken Komatsu Hiromitsu Moriyama 《Journal of General Plant Pathology》2018,84(2):92-103
Magnaporthe oryzae chrysovirus 1 strain A (MoCV1-A) is associated with an impaired growth phenotype of its host fungus, Magnaporthe oryzae. In this report, we assayed the virulence and pathogenicity of MoCV1-A-infected and MoCV1-A-free M. oryzae on rice plants. MoCV1-A infection did not affect virulence-associated fungal traits, such as conidial germination and appressorium formation. However, after punch inoculation of leaves on rice plants, MoCV1-A-infected strain formed smaller lesions than the MoCV1-A-free strain did on all rice varieties tested, showing that MoCV1-A infection resulted in reduced virulence of host fungi in rice plants. In contrast, after spray inoculation of rice seedlings, in some cases, MoCV1-A-infected and MoCV1-A-free strains caused different lesion types (resistance to susceptible, or vice versa) on individual international differential rice varieties. However, we did not find any gain/loss of the fungal avirulence genes by PCR, suggesting that MoCV1-A infection can convert the pathogenicity of the host M. oryzae from avirulence to virulence, or from virulence to avirulence, depending on the rice variety. We also confirmed the correlation of these race conversion events and invasive hyphae growth of the fungi in a leaf sheath inoculation assay. These data suggested that MoCV1-A infection generally confers hypovirulence to the fungal host and could be a driving force to generate physiological diversity, including pathogenic races. 相似文献
110.
Toshiyuki Imaizumi 《Weed Biology and Management》2018,18(2):99-102
Weedy rice is a conspecific weed of cultivated rice (Oryza sativa L.) and is a primary weed of direct‐seeded rice production systems worldwide. The shift in the method of establishing rice, that is, from transplanting to direct seeding, is considered the main reason for the emergence of weedy rice in Asia and, more recently, Japan. Japan has increasingly adopted directly seeded rice, even though this practice remains limited. In this study, I confirmed the relationship between weedy red rice emergence and the adoption of direct‐seeded rice in Japan. Twenty‐seven areas from eight prefectures infested with weedy rice were selected to confirm the relationship between the rice planting method and the emergence of weedy rice. Fields that were severely infested with weedy rice were selected as survey fields for this study. I found no relationship between the method of planting rice and weedy rice emergence. Changes to herbicide application, duration of rice production and hand weeding also contribute to the emergence of weedy rice in transplanted rice fields in Japan. The results of this study indicate that weedy rice could become a serious problem in Japan, even in transplanted rice fields. 相似文献