Construction of a system for exploring mitotic homologous recombination in the genome of Pyricularia oryzae     

Takayuki Arazoe  Shuichi Ohsato  Tsutomu Arie  Katsuyoshi Yoneyama  Shigeru Kuwata 《Journal of General Plant Pathology》2013,79(6):422-430

To investigate mitotic homologous recombination (HR) in Pyricularia oryzae, we created an HR detection system. The system consists of two non-functional enhanced yellow fluorescent protein (YFP) and blasticidin S deaminase (BSD) fusion genes (YFP::BSD). If mitotic HR occurs between the two non-functional genes in the genome, restoration of the functional YFP::BSD gene can be expected. The expression of the functional YFP::BSD gene can be detected by both YFP fluorescence and resistance against BS. When the P. oryzae genome was transformed simultaneously with two non-functional genes, all six lines of transformants with both genes had some portion of their hyphae exhibiting YFP fluorescence and BS resistance during growth. Up to ca. 10 % of conidia harvested from the mycelium of each of the six lines had YFP fluorescence, suggesting that HR consistently occurs during mycelium growth. To determine whether and how the HR-mediated phenotypic changes occurred at the DNA level, we analyzed the genomic DNA of BS-resistant mycelia by PCR-RFLP and sequencing and were able to confirm the existence of a restored functional YFP::BSD gene and a non-functional recombinant gene. Collectively, these findings demonstrate that we established a successful HR detection system for P. oryzae, which can be used for other plant pathogens, and that mitotic HR actually occurs in P. oryzae and constitute the first experimental evidence for mitotic HR in a fungus.  相似文献   

Fecal progestagens to detect and monitor pregnancy in captive female cheetahs (Acinonyx jubatus)     

Adachi I  Kusuda S  Kawai H  Ohazama M  Taniguchi A  Kondo N  Yoshihara M  Okuda R  Ishikawa T  Kanda I  Doi O 《The Journal of reproduction and development》2011,57(2):262-266

The purposes of the present study were to establish a noninvasive monitoring assay of fecal progestagen measurement to detect pregnancy and to identify the components of fecal progestagens in early, middle and late pregnancy in cheetahs. Feces were collected from 7 female cheetahs and analyzed from 30 days before the last copulation to parturition in 9 pregnancies. Blood was collected from one cheetah. Fecal progestagen and serum progesterone concentrations were determined by enzyme immunoassay (EIA). The profiles of the fecal progestagen concentrations were similar to the serum progesterone profile. Fecal progestagen and serum progesterone concentrations remained at the baseline until copulation. In the mean fecal progestagen profile during pregnancy (92.8 ± 0.4 days; from the last copulation to parturition), the concentrations increased 3-4 days after the last copulation and remained high until parturition. To investigate changes in the components of progestagen metabolites in the tripartite periods of gestation, fecal progestagens were analyzed by HPLC-EIA. Marked immunoreactive peaks consistent with 5α-pregnan-3α/β-ol-20-one and 5α-pregnan-3,20-dione and small peaks consistent with 5β-pregnan-3α/β-ol-20-one were detected. There were no distinct difference in the components of progestagens among the first, second and third trimesters of pregnancy. The hormone assay, as an indicator of fecal 5α-reduced pregnanes, is useful for detecting pregnancy and monitoring pregnant luteal activity in cheetahs.  相似文献   

Alleviation of manganese toxicity and manganese-induced iron deficiency in barley by additional potassium supply in nutrient solution     

Shah Alam  Muhammad Hasinur Rahman  Shigeru Kamei  Shigenao Kawai 《Soil Science and Plant Nutrition》2013,59(3):387-392

Barley plants were grown hydroponically at two levels of K (3.0 and 30 mm) and Fe (1.0 and 10 μm) in the presence of excess Mn (25 μm) for 14 d in a phytotron. Plants grown under adequate K level (3.0 mm) were characterized by brown spots on old leaves, desiccation of old leaves, interveinal chlorosis on young leaves, browning of roots, and release of phytosiderophores (PS) from roots. These symptoms were more pronounced in the plants grown under suboptimal Fe level (1.0 p,M) than in the plants grown under adequate Fe level (10 μm). Plants grown in 10 μm Fe with additional K (30 mm) produced a larger amount of dry matter and released less PS than the plants grown under adequate K level (3.0 mm), and did not show leaf injury symptoms and root browning. On the other hand, the additional K supply in the presence of 1.0 μM Fe decreased the severity of brown spots, prevented leaf desiccation, and increased the leaf chlorophyll content, which was not sufficient for the regreening of chlorotic leaves. These results suggested that the additional K alleviated the symptoms of Mn toxicity depending on the Fe concentration in the nutrient solution. The concentration (per g dry matter) and accumulation (per plant) of Mn in shoots and roots of plants grown in 10 μm Fe and 30 mm K were much lower than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that additional K repressed the absorption of Mn. The concentration and accumulation of Fe in the shoots and roots of the plants grown in 10 μm Fe and 30 mm K were higher than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that the additional K increased the absorption of Fe under excess Mn level in the nutrient solution. The release of PS, chlorophyll content, and shoot Fe concentration were closely correlated.  相似文献   

Diurnal variations in absorption and translocation of a ferrated phytosiderophore in barley as affected by iron deficiency     

Shah Alam  Fumihito Akiha  Shigeru Kamei  Shigenao Kawai 《Soil Science and Plant Nutrition》2013,59(3):457-461

The release of phytosiderophore (PS) from roots of Fe-deficient graminaceous plants follows a distinct diurnal rhythm with maximum release rates occurring usually 3 to 4 hours after the onset of light. However, it remains to be determined whether absorption of the PS-Fe³⁺ complex shows a diurnal rhythmicity similar to that of PS release, Barley plants grown with or without 10 µM FeEDTA for 7 days were fed with ferreted PS (10 µM labelled with ⁵⁹Fe) at 4-h intervals to study the diurnal variations in the absorption and transloca tion of ⁵⁹Fe, The absorption of ⁵⁹Fe, irrespective of the Fe nutritional status of the plants, was higher during the day and lower during the night but did not show any peak throughout the day-night cycle. On the other hand, the translocation of ⁵⁹Fe into shoots of Fe-deficient plants was lower than that of Fe-sufficient plants, while the Fe nutritional status of the plants did not affect the absorption of ⁵⁹Fe by roots, The formation of root apoplastic ⁵⁹Fe was lower during the day and higher during the night, regardless of the Fe nutritional status of plants. Our results showed that the absorption of the PS-Fe³⁺ complex by roots did not follow the PS release pattern.  相似文献   

Identification of several rice genes regulated by Si nutrition     

Shuichi Watanabe  Emi Shimoi  Naoko Ohkama  Hiroaki Hayashi  Tadakatsu Yoneyama  Junshi Yazaki 《Soil Science and Plant Nutrition》2013,59(8):1273-1276

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Effectiveness of Phytosiderophore in Absorption and Translocation of 59Iron in Barley in the Presence of Plant-Borne,Synthetic, and Microbial Chelators     

Shah Alam  Shigeru Kamei  Shigenao Kawai 《Journal of plant nutrition》2013,36(10):1709-1722

The mechanisms of iron (Fe) absorption and translocation in plants have received much study because they are the key processes in the supply of Fe to plants. The objective of this research was to study the effectiveness of phytosiderophore (PS) in the absorption and translocation of ⁵⁹Fe in Fe-deficient barley (Hordeum vulgare L. cv. ‘Minorimugi’) plants in the presence of plant-borne, synthetic, or microbial chelators. Plants grown under Fe-deficient conditions in a phytotron at pH 5.5 for 7–18 d were fed with Fe³⁺ (10 μ M labeled with ⁵⁹Fe) in the presence of 10 μ M of different chelators with or without 10 μ M PS for 4 h starting at 2 p.m. (6 h after the onset of light period). The absorption and translocation of ⁵⁹Fe in plants treated with PS and Fe^{3 +} were increased relative to plants fed solely with Fe^{3 +} (control). There was no effect found on absorption and translocation of ⁵⁹Fe in plants treated with EDTA or p-coumarate relative to the control, but a differential increase was observed in ⁵⁹Fe absorption and translocation in plants treated with EDTA or p-coumarate in the presence of PS. In comparison with the control, a decrease in ⁵⁹Fe absorption and translocation was observed in plants treated with HEDTA or EDDHA or FOB, but this decrease was avoided in plants treated with HEDTA or EDDHA or FOB in the presence of PS. The enhancement of ⁵⁹Fe absorption and translocation in plants treated with citrate, and the highest ⁵⁹Fe absorption and translocation in plants treated with citrate and PS, indicated that citrate had an additive effect on Fe absorption and translocation in plants. Our results showed that PS effectively played a role in Fe absorption and translocation in plants in the presence of other chelators. Plants treated with any chelators had lower extracellular ⁵⁹Fe in the roots compared with the control.  相似文献   

Physiological and Mineralogical Properties of Arsenic-Induced Chlorosis in Barley Seedlings Grown Hydroponically     

Molla Rahman Shaibur  Nobuyuki Kitajima  Reiko Sugawara  Toshihito Kondo  S. M. Imamul Huq  Shigenao Kawai 《Journal of plant nutrition》2013,36(2):333-353

The experiment was carried out to investigate the effects of arsenic (As) on the physiological and mineralogical properties of barley (Hordeum vulgare L. cv. ‘Minorimugi’). The plants were grown in nutrient solution treated with 0, 6.7, 33.5, and 67 μ M As (0, 0.5, 2.5, and 5 ppm As, respectively) in the phytotron. Dry matter yield of shoots and roots decreased significantly with the As treatments, indicating that barley plants are As-sensitive and As-toxicity depends on the As concentration in the rooting medium. Necrosis in older leaves and chlorosis symptoms (whitish color) in the fully developed young leaves were observed at the 33.5 and 67 μ M As treatments. Arsenic concentration, accumulation, and translocation increased with the increase of As concentration in the rooting medium. Arsenic was mostly concentrated in roots and a little amount was moved to shoots, indicating that As was not easily translocated to shoots of barley seedlings. Concentrations and accumulations of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), manganese (Mn), zinc (Zn), and copper (Cu) decreased significantly in shoots for 33.5 and 67 μ M As treatments as compared to the 0 μ M As treatment. Concentrations of P, K, Ca, Mg, Mn, and Cu decreased in roots, but Zn concentration increased in roots at 67 μ M As treatment. Accumulations of P, K, Ca, Mg, Mn, Zn, and Cu in roots also decreased significantly at 67 μ M As treatment. Accumulation of P and the cations showed negative relationship with As. Concentration of Fe decreased in shoots at 33.5 and 67 μ M As treatments where chlorosis was induced in the young leaf but increased in roots at 33.5 and 67 μ M As treatments. It was suggested that As might induce iron (Fe)-chlorosis in the plants. Among the micronutrients, Fe translocation was more affected than others by As. Phytosiderophore (PS) accumulation in roots, which is a symptom of Fe-deficiency in grasses, did not change significantly between 0 and 33.5 μ M As treatments; indicating that As-induced chlorosis did not enhance PS accumulation in roots and decreased due to As-toxicity at 67 μ M As treatment.  相似文献   

Studies on the nitrification in soil,with special reference to the population of nitrifier     

Shuichi Ishizawa  Tatsuhiko Matsuguchi 《Soil Science and Plant Nutrition》2013,59(6):30-34

A number of papers have been published on nitrification in soil. The problem of the nitrification has become increasingly important because of the appearance of new forms of nitrogenous fertilizers such as ammonium chloride or urea, etc.  相似文献   

Incorporation of 14C into methionine as an intermediate for phytosiderophore biosynthesis in barley     

Shigenao Kawai  Kiyoshi Kuwajima 《Soil Science and Plant Nutrition》2013,59(4):973-979

In order to verify the precursory role of methionine (Met) in the biosynthesis of the mugineic acid family of phytosiderophores (MAs), feeding experiments of ¹⁴C-Iabeled compounds to barley (Hordeum vulgare L. cv. Minorimugi) roots grown hydroponically were conducted. When both ^l4C-Glucose (Glc) and unlabeled Met were fed to segmented roots, ¹⁴C was incorporated into Met and MAs in the roots. Molar-radioactivity of Met was higher than that of the amino-butanoic-acid unit in MAs in the roots. When ^l4C-Glc and unlabeled homoserine (Hse) were fed to decapitated roots, l4C was incorporated into Met but not into Hse. Therefore, it was considered that Hse might not be a major precursor of MAs. In addition, ¹⁴C was incorporated into Met and MAs in the roots when both ¹⁴C-glycerol (Gol) and unlabeled Met were fed to segmented roots. It is suggested that MAs may be synthesized from Glc via Met, bypassing Hse, and that the MAs biosynthesis may involve an unknown pathway associated with Gol and leading to Met.  相似文献   

A next-generation-sequencing approach to diagnose viral diseases of greenhouse ranunculus     

Hayahi  Saki  Ota  Emi  Kurogi  Shuichi  Matsushita  Yosuke  Kanno  Yoshiaki  Takeshita  Minoru 《Journal of General Plant Pathology》2022,88(2):134-137

Journal of General Plant Pathology - Viruses that infect Ranunculus asiaticus L., a globally important greenhouse ornamental, have consistently caused serious damage in Japan. We coupled...  相似文献