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Kyoko Sugawara Misako Himeno Takuya Keima Yugo Kitazawa Kensaku Maejima Kenro Oshima Shigetou Namba 《Journal of General Plant Pathology》2012,78(6):389-397
Phytoplasmas are plant pathogenic bacteria that infect more than 700 plant species. Because phytoplasma-resistant cultivars are not available for the vast majority of crops, the most common practice to prevent phytoplasma diseases is to remove infected plants. Therefore, developing a rapid, accurate diagnostic method to detect a phytoplasma infection is important. Here, we developed a phytoplasma detection assay based on loop-mediated isothermal amplification (LAMP) by targeting the groEL gene and 16S rDNA. We designed 19 primer sets for the LAMP assay and evaluated their amplification efficiency, sensitivity, and spectra to select the most suitable primer sets to detect Candidatus Phytoplasma asteris. As a result, DNA was efficiently amplified by one of the primer sets targeting the groEL gene, and LAMP assay sensitivity with this primer set was 10-fold higher than that of the polymerase chain reaction. Moreover, the groEL gene was successfully amplified from several strains of Ca. Phytoplasma asteris by this primer set, indicating that the groEL gene can be used as a LAMP assay target gene for a broad range of phytoplasma strains. Additionally, a simple DNA extraction method that omits the homogenizing and phenol extraction steps was combined with the LAMP assay to develop a simple, rapid, and convenient diagnostic method for detecting phytoplasma. 相似文献
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Hiroshi Fukayama Takashi Kobara Keita Shiomi Ryutaro Morita Daisuke Sasayama Tomoko Hatanaka 《Plant Production Science》2013,16(2):296-300
ABSTRACT
Overexpression of Rubisco small subunit (RbcS) of C4 plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (k cat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C4 plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS (NgRbcS) and guinea grass RbcS (GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The k cat and K m for CO2 (Kc) of Rubisco were increased in all transgenic lines. Interestingly, the k cat was markedly higher in NgRbcS homozygous line, whereas K c was notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C4 RbcS are effective approaches to alter the catalytic properties of Rubisco in rice. 相似文献
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Katsuhiro Tokugawa Dan Aoki Ryutaro Asai Yasuyuki Matsushita Masao Ishiguro Yasufumi Noda Kazuhiko Fukushima 《Journal of Wood Science》2017,63(3):281-287
In order to understand the kraft pulp decolouring mechanism on using a nonionic detergent, the pulp washing process and the resulting pulp handsheets were investigated by examining the brightness, kappa number, thioacidolysis product yield, and dewatering efficiency in the pressing sheet making process. The pulp decolouring could be attributed to a decrease in the lignin content and an improvement in the dewatering efficiency. Furthermore, the detergent distribution in the aqueous pulp suspension obtained during the pulp washing process was visualised using cryo-time-of-flight secondary ion mass spectrometry/scanning electron microscopy (cryo-TOF-SIMS/SEM). The detergent was clearly observed at the transverse surface of the pulp fibre cell wall and was also detected in the lumen of the fibres, suggesting the permeation of the detergent into the pulp fibre cell wall. Based on these results, the pulp decolouring mechanism can be proposed as follows: the detergent permeates into the pulp fibre cell wall and promotes the solution-exchange between the inside and the outside parts of the fibre cell wall, finally washing away the chromophoric substances such as lignin and its degradation products owing to the enhanced dewatering efficiency. 相似文献
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This study describes a successful method of somatic embryogenesis and genetic transformation using immature cotyledons of Prunus mume. Immature cotyledons from four different developmental stages of eight different P. mume cultivars were used for the experiments to optimize somatic embryogenesis and genetic transformation protocols. Somatic embryogenesis was induced when the explants were cultured on somatic embryo inducing medium consisting of MS basic medium supplemented with 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 μM 6-benzyladenine (BA). They were cultured for 30 days and then transferred to somatic embryo propagation medium containing 0.1 μM α-naphthaleneacetic acid (NAA) and 5 μM BA. It appeared that the developmental stage of the immature cotyledons used as explants was the most important factor for somatic embryogenesis; higher frequencies of somatic embryogenesis were observed when the immature cotyledons were less than 5 mm in length regardless of cultivars. For genetic transformation, the immature cotyledons were inoculated with Agrobacterium tumefaciens EHA101 harbouring a binary plasmid vector with neomycin phosphotransferase II and an intron-interrupted β-glucuronidase gene under the control of cauliflower mosaic virus 35S promoter, and three transgenic plant lines were obtained from inoculated “Sirakaga” immature cotyledons. Transgenic somatic embryos and shoots were selected using 25 mg l−1 kanamycin. Integration of transgenes in the genome of GUS-positive putative transgenic shoots was confirmed by PCR and Southern blot analyses. 相似文献
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Yoshikawa Ryutaro Maeda Atsushi Ueno Yoshihito Sakai Hiroki Kimura Shintaro Sawadaishi Tomohiro Kohgo Satoru Yamada Kohei Mori Takashi 《Veterinary research communications》2022,46(2):447-457
Canine hemangiosarcoma (HSA) has an extremely poor prognosis, making it necessary to develop new systemic treatment methods. MicroRNA-214 (miR-214) is one of many microRNAs (miRNA) that can induce apoptosis in HSA cell lines. Synthetic miR-214 (miR-214/5AE), which showed higher cytotoxicity and greater nuclease resistance than mature miR-214, has been developed for clinical application. In this study, we evaluated the effects of miR-214/5AE on stage 2 HSA in a mouse model. Mice intraperitoneally administered with miR-214/5AE (5AE group) had significantly fewer intraperitoneal dissemination tumor foci (median number: 72.5 vs. 237.5; p?<?0.05) and a lower median foci weight (0.26 g vs. 0.61 g; p?<?0.05). Mice in the 5AE group had increased expression of p53 and cleaved caspase-3, and a significantly lower proportion of Ki-67-positive cells, than those in the non-specific miR group. Notably, no significant side effects were observed. These results indicate that intraperitoneal administration of miR-214/5AE exhibits antitumor effects in an intraperitoneal dissemination mouse model of HSA by inducing apoptosis and suppressing cell proliferation. These results provide a basis for future studies on the antitumor effect of miR-214/5AE for HSA.
相似文献9.
Kensaku Maejima Misako Himeno Osamu Netsu Kazuya Ishikawa Tetsuya Yoshida Naoko Fujita Masayoshi Hashimoto Ken Komatsu Yasuyuki Yamaji Shigetou Namba 《Journal of General Plant Pathology》2014,80(2):176-183
Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains. 相似文献
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Most Prunus fruit tree species exhibit a homomorphic gametophytic self-incompatibility (GSI) system, in which specificity of self/nonself-recognition is controlled by products encoded within the S locus. In the pollination event, a self-incompatibility (SI) reaction is triggered when the same “S allele” specificity is expressed in both the pollen and pistil. During the last two decades, much progress has been made in our understanding of the molecular basis of the gametophytic self-incompatibility system in Prunus. Identification of the pistil S and pollen S determinants led to the development of PCR-based S genotyping and marker-assisted selection for self-compatible (SC) individuals. Molecular and genetic analyses of Prunus SC S haplotypes and polyploid sour cherry (Prunus cerasus) reveal the possible existence of a distinct SI/SC recognition mechanism in the S-RNase-based GSI system of Prunus. This review summarizes the current molecular knowledge of the S-RNase-based GSI system in Prunus with reference to data collected for S-RNase-based GSI in other plants and its potential usefulness in SC breeding. 相似文献