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41.
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
42.
Shiro Fukuta Reiko Takahashi Satoru Kuroyanagi Noriyuki Miyake Hirofumi Nagai Hirofumi Suzuki Fujio Hashizume Tomoko Tsuji Hiromi Taguchi Hideki Watanabe Koji Kageyama 《European journal of plant pathology / European Foundation for Plant Pathology》2013,136(4):689-701
A loop-mediated isothermal amplification (LAMP) reaction with a primer set designed from the rDNA ITS sequence of P. aphanidermatum was developed. Results of a specificity test using 57 strains of Pythium spp. indicated that the LAMP assay gave no cross reactions in other 39 Pythium species, 11 strains of Phytophthora spp. and eight other soil borne pathogens. The detection limit was 10 fg of genomic DNA, which was ten times the sensitivity of the polymerase chain reaction. The LAMP assay was applied to hydroponic solution samples from tomato fields, and the results were compared to those of the conventional plating method. LAMP was observed to be effective for the specific detection of P. aphanidermatum. Furthermore, P. aphanidermatum was detected directly in tomato roots infected with P. aphanidermatum without DNA extraction. The LAMP method established in this study is a simple, sensitive and rapid tool for the detection of P. aphanidermatum. 相似文献
43.
Mizuho TASHIRO Yuki HOSOKAWA Hiromi AMAO Atsushi TOHEI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2020,82(12):1757
Hypothermia during anesthetic events is a common adverse effect of anesthesia in laboratory animals. In particular, small rodents such as mice is susceptible to hypothermia during anesthetic events. Therefore, the animals will need additional thermal support by external heating devices during and after anesthesia. In general, the time of recovery from anesthesia is typically longer in case of injectable anesthesia rather than inhalant anesthesia. However, the durations of thermal support have been almost limited to 1 hr from administration of anesthesia in general. Our study objectives are two-fold: 1) to compare the levels of hypothermia induced by injectable anesthesia with medetomidine-midazolam-butorphanol (MMB) and inhalant anesthesia with isoflurane (ISO); 2) to find the adequate durations of thermal support for preventing hypothermia induced by their anesthesia in mice. Adult male ICR mice were anesthetized during 40 min without and with the thermal support for 1 (both anesthetic groups), 2, 3, and 5 hr (in MMB group). Without thermal support, the decrease of body temperature in MMB group were more severe than that in ISO group. The durations of thermal support completely prevented hypothermia at 5 hr-support in MMB group and that at 1 hr-support in ISO group. However, the other short durations did not prevent hypothermia at 1, 2 and 3 hr-support in MMB group. These results suggest that the mice should be received thermal support over 5 hr after injection of MMB anesthesia to prevent hypothermia. 相似文献
44.
Makoto Shinohara Chihiro Aoyama Kazuki Fujiwara Atsunori Watanabe Hiromi Ohmori Yoichi Uehara 《Soil Science and Plant Nutrition》2013,59(2):190-203
Hydroponics is an excellent technique for the cultivation of vegetable crops and other plants, but organic fertilizers cannot be used in conventional hydroponic systems, which generally use only inorganic fertilizers, because organic compounds in the hydroponic solutions generally have phytotoxic effects that lead to poor plant growth. Few microorganisms are present in hydroponic solutions to mineralize the organic compounds into inorganic nutrients. In this article a novel and practical hydroponic culture method that uses microorganisms to degrade organic fertilizer in the hydroponic solution has been developed. Soil microorganisms were cultured by regulating the amounts of organic fertilizer and inoculum, with moderate aeration. The microorganisms mineralized organic nitrogen via ammonification and nitrification into nitrate at an efficiency of 97.6%. The culture solution containing the microorganisms was usable as a hydroponic solution, and organic fertilizer could be directly added to it during vegetable cultivation. Vegetables grew well in the organic hydroponic system. Organic hydroponics based on this method is therefore a practical tool for the utilization of organic sources of fertilizer. 相似文献
45.
Toshihiro Watanabe Mitsuru Osaki Hiromi Yano Idupulapati M. Rao 《Journal of plant nutrition》2013,36(7):1243-1255
Many tropical forage grasses and legumes grow well in acid soils, adapting to excess aluminum (Al) and phosphorus (P) starvation stresses by using mechanisms that are still unclear. To determine these mechanisms, responses to Al toxicity and P starvation in three tropical forages were studied: two grasses, Brachiaria hybrid cv. ‘Mulato’ (B. ruziziensis clone 44-06 × B. brizantha cv. ‘Marandú’) and Andropogon gayanus, and one legume, Arachis pintoi. The tropical grasses tolerated high levels of Al toxicity and P starvation, with the Brachiaria hybrid maintaining very low levels of Al concentration in shoots. 27Al Nuclear Magnetic Resonance spectroscopy (NMR) analysis revealed that, in the Brachiaria hybrid, Al makes complexes with some ligands such as organic-acid anions in the root symplast. The forages probably adapted to P starvation through high P-use efficiency. These experiments provide the first direct evidence we know of that organic acid anions within root tissue help detoxify Al in non-accumulator species such as the Brachiaria hybrid. 相似文献
46.
Hiromi Yokota Ikuyo Takamura Fumiko Ishikawa Miwako Ohta Shigeki Konishi 《Soil Science and Plant Nutrition》2013,59(2):457-461
It is well known that when inorganic elements such as boron (B), calcium, cobalt, and manganese are added to media in combination or alone, they stimulate the germination and/or tube growth of various kinds of pollen (Schmücker 1933; Loo and Hwang 1944; Yamada 1958; Brewbaker and Kwack 1963). Among these elements, the stimulatory effect ofB is the most effective and generally recognized. Aluminum (Al) belongs along with B to Group IIIa in the periodic table, and Fähnrich and Ultlich (1964) reported that Al inhibited the germination and growth of the pollen tube of Petunia hybrida and Antirrhinum majus. However, stimulatory effects of Al on pollen germination or pollen tube growth had not been previously reported. 相似文献
47.
Ch unlan Lian Yoshihisa Oi wake Hiromi Yokota Gang Wang Shigeki Konishi 《Soil Science and Plant Nutrition》2013,59(4):695-700
Aluminum (Al) toxicity is a major factor limiting yield production on acid soils (Foy 1983). The initial symptom of Al toxicity in many plants is manifested by the inhibition of root elongation (Ownby and Popham 1990; Llugany et al. 1994; Sasaki et al. 1994; Horst et al. 1997), which occurs during a very short period of time after exposure to Al (Llugany et al. 1994; Staß and Horst 1995). In a large number of recent reports, it was shown that the root apex plays a major role in the Al-sensitivity and response mechanisms (Zhang et al. 1994; Sasaki et al. 1997; Sivaguru and Horst 1998). However, it is interesting to note that stimulatory effects of Al on the growth of plants have also been reported in some studies (Chenery 1955; Konishi et al. 1985; Huang and Bachelard 1993; Osaki et al. 1997). In tea plant (Camellia sinensis L.) a stimulatory effect of Al on the growth was also demonstrated in some experiments, using intact plant (Chenery 1955; Konishi et al. 1985), cultured roots (Tsuji et al. 1994), and pollen tubes (Yokota et al. 1997). The growth of tea roots was typically more stimulated than that of shoots by Al (Konishi et al. 1985). It was assumed that Al effects might be due to the amelioration of phosphorus absorption (Konishi et al. 1985), secretion of malic acid from roots to dissolve aluminum phosphate in the rhizosphere (Jayman and Sivasubramaniam 1975), stimulation of growth of microorganisms on the root surface (Konishi 1990) or replacement of some functions of boron (Konishi 1992; Yokota et al. 1997). However, the stimulatory effects of Al on tea plant growth have not yet been el ucidated. The formation of callose (1,3-β-glucan) has been reported as a common plant response to a variety of stresses, as well as mechanical, biophysical, chemical, and biological injury (Jaffe and Leopold 1984; Zhang et al. 1994). Increased synthesis of callose has been observed upon exposure to excess amounts of some elements, such as boron (McNairn and Currier 1965), cobalt, nickel, zinc (Peterson and Rauser 1979), and manganese (Wissemeier and Horst} 1987, 1992). Callose synthesis was also induced by Al in the roots of Triticum aestivum (Zhang et al. 1994) and Zea mays (Horst et al. 1997; Sivaguru and Horst 1998), suspension-cultured cells of Glycine max (Staß and Horst 1995), and protoplasts of Avena sativa (Schaeffer and Walton 1990) and Zea mays (Wagatsuma et al. 1995). Induction of callose synthesis in roots seems to be a very rapid physiological indicator of Al-induced injury or genotypical differences in Al sensitivity (Wissemeier and Horst 1992; Zhang et al. 1994; Horst et al. 1997). Nevertheless, Al-induced callose synthesis in tea plant, whose growth is stimulated by suitable Al concentrations, has not been described yet. Therefore, to elucidate the physiological basic effects of Al on tea plant, callose synthesis affected by Al in the root tips of intact plants was analyzed in the present study. 相似文献
48.
Kenji Kanazawa Kyoko Higuchi Hiromi Nakanishi Naoko Kishi-Nishizawa Satoshi Mori 《Soil Science and Plant Nutrition》2013,59(4):717-721
Under iron deficient conditions, graminaceous plants secrete mugineic acid family phytosiderophores (MAs) from their roots to dissolve sparingly soluble iron compounds in the rhizosphere, and take up iron in the form of an Fe3+-MAs complex (Takagi 1976). A good correlation has been reported between the tolerance of Fe-deficiency and the amount of secreted MAs (Takagi 1993). Therefore, by using the genes involved in MAs biosynthesis, molecular breeding might produce transgenic plants tolerant to Fe-deficiency with a high level of MAs secretion. The biosynthetic pathway of MAs from L-methionine has been clarified (Fig. 1) and the enzymes participating in this process are now being investigated to isolate the genes responsible. Nicotianamine aminotransferase (NAAT) catalyzes the amino group transfer between nicotianamine (NA) and 2-oxoglutaric acid (Fig. 1). In order to purify NAAT, enzyme assay methods for NAAT have been developed and modified (Shojima et al. 1990; Ohata et al. 1993; Kanazawa et al. 1994). Some characteristics of NAAT have been reported using these enzyme assay methods (Kanazawa et al. 1994, 1995). Here, we further investigate some characteristics of this enzyme to improve the enzyme assay method, namely 1) the effect of K+ and Mg2+ on NAAT activity in vitro, and 2) the direct influence of MAs, Fe3+, and Fe2+ on NAAT activity. In addition, based on these results, the induction of enzyme activity by Fe-deficiency and suppression of the activity by Fe-resupply was investigated, by applying the new enzyme assay method. 相似文献
49.
Jun HAKOZAKI Sho KUZUKAMI Asako HARAGUCHI Kazuhiko NAKAYAMA Kodai KUSAKISAKO Noboru KUDO Hiromi IKADAI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(6):927
The definitive hosts of Metagonimus hakubaensis are reported to be hamsters, rats, mice, dogs, cats, chickens, and quails in experimental infection and Japanese water shrews in natural infection. Here we report that raccoon dogs are new natural definitive hosts of M. hakubaensis, based on morphological and molecular analyses of Metagonimus flukes collected from the host species from Aomori Prefecture, Japan. Moreover, M. hakubaensis recovered from raccoon dogs showed higher fecundity than those recovered from Japanese water shrews. Therefore, raccoon dogs were considered as a more suitable natural definitive host of M. hakubaensis than Japanese water shrews. 相似文献
50.
Increase in Iron and Zinc Concentrations in Rice Grains Via the Introduction of Barley Genes Involved in Phytosiderophore Synthesis 总被引:1,自引:1,他引:0
Hiroshi Masuda Motofumi Suzuki Kendi Claudio Morikawa Takanori Kobayashi Hiromi Nakanishi Michiko Takahashi Masahiko Saigusa Satoshi Mori Naoko K. Nishizawa 《Rice》2008,1(1):100-108
Increasing the iron (Fe) and zinc (Zn) concentrations of staple foods, such as rice, could solve Fe and Zn deficiencies, which are two of the most serious nutritional problems affecting humans. Mugineic acid family phytosiderophores (MAs) play a very important role in the uptake of Fe from the soil and Fe transport within the plant in graminaceous plants. To explore the possibility of MAs increasing the Fe concentration in grains, we cultivated three transgenic rice lines possessing barley genome fragments containing genes for MAs synthesis (i.e., HvNAS1, HvNAS1, and HvNAAT-A and HvNAAT-B or IDS3) in a paddy field with Andosol soils. Polished rice seeds with IDS3 inserts had up to 1.40 and 1.35 times higher Fe and Zn concentrations, respectively, compared to non-transgenic rice seeds. Enhanced MAs production due to the introduced barley genes is suggested to be effective for increasing Fe and Zn concentrations in rice grains. 相似文献