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1.
2.
It has been pointed out that tea (Camellia sinensis (L.) O. Kuntze) prefers ammonium (NH 4 + ) over nitrate (NO 3 ? ) as an inorganic nitrogen (N) source. 15N studies were conducted using hydroponically grown tea plants to clarify the characteristics of uptake and assimilation of NH 4 + and NO 3 ? by tea roots. The total 15N was detected, and kinetic parameters were calculated after feeding 15NH 4 + or 15NO 3 ? to tea plants. The process of N assimilation was studied by monitoring the dynamic 15N abundance in the free amino acids of tea plant roots by GC-MS. Tea plants supplied with 15NH 4 + absorbed significantly more 15N than those supplied with 15NO 3 ? . The kinetics of 15NH 4 + and 15NO 3 ? influx into tea plants followed a classic biphasic pattern, demonstrating the action of a high affinity transport system (HATS) and a low affinity transport system (LATS). The V max value for NH 4 + uptake was 54.5 nmol/(g dry wt min), which was higher than that observed for NO 3 ? (39.3 nmol/(g dry wt min)). KM estimates were approximately 0.06 mM for NH 4 + and 0.16 mM for NO 3 ? , indicating a higher rate of NH 4 + absorption by tea plant roots. Tea plants fed with 15NH 4 + accumulated larger amounts of assimilated N, especially glutamine (Gln), compared with those fed with 15NO 3 ? . Gln, Glu, theanine (Thea), Ser, and Asp were the main free amino acids that were labeled with 15N under both conditions. The rate of N assimilation into Thea in the roots of NO 3 ? -supplied tea plants was quicker than in NH 4 + -supplied tea plants. NO 3 ? uptake by roots, rather than reduction or transport within the plant, seems to be the main factor limiting the growth of tea plants supplied with NO 3 ? as the sole N source. The NH 4 + absorbed by tea plants directly, as well as that produced by NO 3 ? reduction, was assimilated through the glutamine synthetase-glutamine oxoglutarate aminotransferase pathway in tea plant roots. The 15N labeling experiments showed that there was no direct relationship between the Thea synthesis and the preference of tea plants for NH 4 + .  相似文献   

3.
In plants of wheat (Triticum aestivum L.) grown in the media with nitrate (NO 3 ? plants), ammonium (NH 4 + plants), and without nitrogen (N-deficient plants), the response to oxidative stress induced by the addition of 300 mM NaCl to the nutrient solution was investigated. Three-day-long salinization induced chlorophyll degradation and accumulation of malondialdehyde (MDA) in the leaves. These signs of oxidative stress were clearly expressed in NO 3 ? and N-deficient plants and weakly manifested in NH 4 + plants. In none of the treatments, salinization induced the accumulation of MDA in the roots. Depending on the conditions of N nutrition, salt stress was accompanied by diverse changes in the activity of antioxidant enzymes in the leaves and roots. Resistance of leaves of NH 4 + plants to oxidative stress correlated with a considerable increase in the activities of ascorbate peroxidase and glutathione reductase. Thus, wheat plants grown on the NH 4 + -containing medium were more resistant to the development of oxidative stress in the leaves than those supplied with nitrate.  相似文献   

4.
Water stress is a primary limitation on plant growth. In previous studies, it has been found that ammonium enhances the tolerance of rice plants to water stress, but how water is related to nitrogen form and water stress remains unknown. To study the effects of nitrogen form (NH 4 + , NO 3 ? , and a mixture of NH 4 + and NO 3 ? ) on the growth and water absorption of rice (Oryza sativa L.) seedlings, a hydroponic experiment with water stress, simulated by the addition of polyethylene glycol (PEG, 10% w/v, MW 6000), was conducted in a greenhouse. The results showed that, compared with non-water stress, under water stress, the fresh weight of rice seedlings increased by 14% with NH 4 + nutrition, whereas it had decreased by about 20% with either NO 3 ? or mixed nitrogen nutrition. No significant difference was found in the transpiration rate of excised shoots or in xylem exudation of excised roots in NH 4 + supply between the two water situations, whereas xylem flow decreased by 57% and 24% under water stress in NO 3 ? and mixed nutrition, and root hydraulic conductivity decreased by 29% and 54% in plants in NH 4 + and NO 3 ? nutrition conditions, respectively. Although water absorption ability decreased in both NH 4 + and NO 3 ? nutrition, aquaporin activity was higher in NH 4 + than in NO 3 ? nutrition, regardless of water stress. We conclude that NH 4 + nutrition can improve water handling in rice seedlings and subsequently enhance their resistance to drought.  相似文献   

5.
This investigation was prompted by the findings that (1) dibucaine-resistant homozygotes and heterozygotes for plasmacholinesterase also exhibit resistance to fluoride inhibition, (2) the differentiation of dibucaine-resistant from the fluoride-resistant genotypes is ambiguous with the method of Harris and Whittaker, (3) the plasmacholinesterase inhibition by Na fluoride (FN) is markedly influenced by the temperature. Therefore, we modified their method by increasing (1) the temperature of the reaction from 25C to 37C and (2) the concentration of Na fluoride from 5.0×10?5 m to 2.5×10?4 m. With this method, genetically normal individuals have a mean FN±sd=77.0±3.22 while atypical dibucaine-resistant homozygotes have a mean FN±sd=43.0±10.0 and atypical dibucaine-resistant heterozygotes 67.0±5.37. Since a linear correlation was observed between DN and FN by our new method, a fluoride number 2 sd lower than the predicted FN from the DN can distinctly identify the fluoride-resistant plasmacholinesterase genotype E 1 f .  相似文献   

6.
The effect of nitrogen starvation on the NO3-dependent induction of nitrate reductase (NR) and nitrite reductases (NIR) has been investigated in the halophilic alga Dunaliella salina. When D. salina cells previously grown in a medium with NH 4 + as the only nitrogen source (NH 4 + -cells) were transferred into NO 3 ? medium, NR was induced in the light. In contrast, when cells previously grown in N-free medium were transferred into a medium containing NO 3 ? , NR was induced in light or in darkness. Nitrate-dependent NR induction, in darkness, in D. salina cells previously grown at a photon flux density of 500 umol · m?2 s?1 was observed after 4 h preculture in N-free medium, whilst in cells grown at 100 umol · m?2 s?1 NR induction was observed after 7–8 h. An inhibitor of mRNA synthesis (6-methylpurine) did not inhibit NO 3 ? -induced NR synthesis when the cells, previously grown in NH 4 + medium, were transferred into NO 3 ? medium (at time 0 h) after 4-h-N starvation. However, when 6-methylpurine was added simultaneously with the transfer of the cells from NH 4 + to NO 3 ? medium (at time 0 h), NO 3 ? induced NR synthesis was completely inhibited. The activity of NIR decreased in N-starved cells and the addition of NO 3 ? to those cells greatly stimulated NIR activity in the light. The ability to induce NR in darkness was observed when glutamine synthetase activity reached its maximal level during N starvation. Although cells grown in NO 3 ? medium exhibited high NR activity, only 0.33% of the total NR was found in intact chloroplasts. We suggest that the ability, to induce NR in darkness is dependent on the level of N starvation, and that NR in D. salina is located in the cytosol. Light seems to play an indirect regulatory role on NO 3 ? uptake and NR induction due to the expression of NR and NO 3 ? -transporter mRNAs.  相似文献   

7.
The effective cross section for the H 2 + + H 2 + → H 3 + + p reaction in the energy range 5.7–11.5 eV is measured by the split beam method. The cross-section maximum at an energy of ~8 eV is related to the production of the H 4 ++ compound system. The reaction threshold W thr ≈5 eV provides evidence in favor of the classical model with the H 2 + ion charge fixed on one of the two nuclei during the entire collision event.  相似文献   

8.
9.

Aims

The objective of this study was to determine the relative importance of transpirational pull, Se speciation, sulfate and species on Se accumulation by plants, in order to determine which of these factors must be considered in the future development of models to predict Se accumulation by plants.

Methods

Seedlings of durum wheat (Triticum turgidum L. var durum cv ‘Kyle’) and spring canola (Brassica napus L. var Hyola 401) were grown hydroponically and exposed to SeO 4 2- (selenate) with or without SO 4 2- (sulfate), or to HSeO 3 - (biselenite) under different transpiration regimes altered through ‘low’ (~50%) or ‘high’ (~78%) relative humidity (RH). Plants were harvested after 0, 8, 16, or 24?h exposures, digested, and analyzed for Se by GFAAS.

Results

Accumulation and distribution of Se by plants is dependent on plant species, Se speciation in the nutrient solution, SO 4 2- competition, and transpiration regimes. Canola accumulated and translocated more Se than wheat. In wheat and canola, the greatest accumulation and translocation of Se occurred when plants were exposed to SeO 4 2- without SO 4 2- compared to solutions of SeO 4 2- with SO 4 2- or HSeO 3 2- . Wheat plants exposed to SeO 4 2- and SO 4 2- had an increased Se accumulation and translocation under increased transpiration rates than when exposed to SeO 4 2- without SO 4 2- or HSeO 3 2- . On the other hand, increases in transpiration increased the translocation of Se to canola shoots when exposed to HSeO 3 - more than any other treatments.

Conclusions

Overall, our results suggest that plant species is the most important factor influencing Se accumulation and translocation, but that these endpoints can be modified by climate and specific soil Se or S content. Models to predict accumulation of Se by plants must consider all of these factors to accurately calculate the mechanisms of uptake and translocation.  相似文献   

10.
Labeled nitrogen (15?N) was applied to a soil-based substrate in order to study the uptake of N by Glomus intraradices extraradical mycelium (ERM) from different mineral N (NO 3 ? vs. NH 4 + ) sources and the subsequent transfer to cowpea plants. Fungal compartments (FCs) were placed within the plant growth substrate to simulate soil patches containing root-inaccessible, but mycorrhiza-accessible, N. The fungus was able to take up both N-forms, NO 3 ? and NH 4 + . However, the amount of N transferred from the FC to the plant was higher when NO 3 ? was applied to the FC. In contrast, analysis of ERM harvested from the FC showed a higher 15?N enrichment when the FC was supplied with 15NH 4 + compared with 15NO 3 ? . The 15?N shoot/root ratio of plants supplied with 15NO 3 ? was much higher than that of plants supplied with 15NH 4 + , indicative of a faster transfer of 15NO 3 ? from the root to the shoot and a higher accumulation of 15NH 4 + in the root and/or intraradical mycelium. It is concluded that hyphae of the arbuscular mycorrhizal fungus may absorb NH 4 + preferentially over NO 3 ? but that export of N from the hyphae to the root and shoot may be greater following NO 3 ? uptake. The need for NH 4 + to be assimilated into organically bound N prior to transport into the plant is discussed.  相似文献   

11.
The effective cross section for the H 2 + +H 2 + → H 3 + +p reaction in the energy range 5.7–11.5 eV is measured by the split beam method. The maximum of the cross section at an energy of ~8 eV is related to the production of the H 4 ++ compound system. The reaction threshold W thr≈5 eV provides evidence in favor of the classical model of the H 2 + ion with the charge fixed on one of the nuclei throughout the collision event.  相似文献   

12.

Aims

Alkali stress (AS) is an important agricultural contaminant and has complex effects on plant metabolism, specifically root physiology. The aim of this study was to test the role of nitrogen metabolism regulation in alkali tolerance of rice variety 'Nipponbare'.

Methods

In this study, the rice seedlings were subjected to salinity stress (SS) or AS. Growth, the contents of inorganic ions, NH 4 + -nitrogen (free amino acids), and NO 3 ? -nitrogen in the stressed seedlings were then measured. The expression of some critical genes involved in nitrogen metabolism were also assayed to test their roles in the regulation of nitrogen metabolism during adaptation of rice variety 'Nipponbare' to AS.

Results

AS showed a stronger inhibiting effect on rice variety 'Nipponbare' growth than SS. AS may have more complex effects on nitrogen metabolism than SS.

Conclusions

Effects of AS on the nitrogen metabolism of rice variety 'Nipponbare' mainly comprised two mechanisms. Firstly, in roots, AS caused the reduction of NO 3 ? content, which caused two harmful consequences, the large downregulation of OsNR1 expression and the subsequent reduction of NH 4 + production in roots. On the other hand, under AS (pH, 9.11), almost all the NH 4 + was changed to NH3, which caused a severe deficiency of NH 4 + surrounding the roots. Both events might cause a severe deficiency of NH 4 + in roots. Under AS, the increased expression of several OsAMT family members in roots might be an adaptative response to the reduction of NH 4 + content in roots or the NH 4 + deficiency in rhizosphere. Also, the down-regulation of OsNADH-GOGAT and OsGS1;2 in roots might be due to NH 4 + deficiency in roots. Secondly, in shoots, AS caused a larger acuumulatiuon of Na+, which possibly affected photorespiration and led to a continuous decrease of NH 4 + production in shoots, and inhibited the expression of OsFd-GOGAT and OsGS2 in chloroplasts.  相似文献   

13.

Background and aims

Plant physiological traits and their relation to soil N availability was investigated as regulators of the distribution of understory shrub species along a slope in a Japanese cedar (Cryptomeria japonica) plantation in central Japan.

Methods

At the study site, previous studies demonstrated that both net and gross soil nitrification rates are high on the lower slope and there are dramatic declines in different sections of the slope gradient. We examined the distributions of understory plant species and their nitrate (NO 3 ? -N) use traits, and compared the results with the soil traits.

Results

Our results show that boundaries between different dominant understory species correspond to boundaries between different soil types. Leucosceptrum stellipilum occurs on soil with high net and gross nitrification rates. Hydrangea hirta is dominant on soil with high net and low gross nitrification rates. Pieris japonica occurs on soil with very low net and gross nitrification rates. Dominant understory species have species-specific physiological traits in their use of NO 3 ? -N. Pieris japonica lacks the capacity to use NO 3 ? -N as a N source, but other species do use NO 3 ? -N. Lindera triloba, whose distribution is unrelated to soil NO 3 ? -N availability, changes the extent to which it uses NO 3 ? -N in response to soil NO 3 ? -N availability.

Conclusions

Our results indicate that differences in the physiological capabilities and adaptabilities of plant species in using NO 3 ? -N as a N source regulate their distribution ranges. The identity of the major form of available soil N is therefore an environmental factor that influences plant distributions.  相似文献   

14.
Growth of 2-month-old nonnodulatedHippophaë rhamnoides seedlings supplied with combined N was compared with that of nodulated seedlings grown on zero N. Plant growth was significantly better with combined N than with N2 fixation and, although not statistically significant for individual harvests, tended to be highest in the presence of NH 4 + , a mixture of NH 4 + and NO 3 ? producing the highest yields. Growth was severely reduced when solely dependent on N2 fixation and, unlike the combined-N plants, shoot to root ratios had only slightly increased after an initial decrease. An apparently insufficient nodule mass (nodule weight ratio <5 per cent) during the greater part of the experimental period is suggested as the main cause of the growth reduction in N2-fixing plants. Thein vivo nitrate reductase activity (NRA) of NO 3 ? dependent plants was almost entirely located in the roots. However, when grown with a combination of NO 3 ? and NH 4 + , root NRA was decreased by approximately 85 per cent.H. rhamnoides demonstrated in the mixed supply a strong preference for uptake of N as NH 4 + , NO 3 ? contributing only for approximately 20 per cent to the total N assimilation. Specific rates of N acquisition and ion uptake were generally highest in NO 3 ? +NH 4 + plants. The generation of organic anions per unit total plant dry weight was approximately 40 per cent less in the NH 4 + plants than in the NO 3 ? plants. Measured extrusions of H+ or OH? (HCO 3 ? ) were generally in good agreement with calculated values on the basis of plant composition, and the acidity generated with N2 fixation amounted to 0.45–0.55 meq H+. (mmol Norg)?1. Without acidity control and in the presence of NH 4 + , specific rates of ion uptake and carboxylate generation were strongly depressed and growth was reduced by 30–35 per cent. Growth of nonnodulatedH. rhamnoides plants ceased at the lower pH limit of 3.1–3.2 and deterioration set in; in the case of N2-fixing plants the nutrient solution pH stabilized at a value of 3.8–3.9 without any apparent adverse effects upon plant performance. The chemical composition of experimental and field-growing plants is being compared and some comments are made on the nitrogen supply characteristics of their natural sites.  相似文献   

15.
Thylakoids were isolated from the leaves of three different plants (Pisum sativium L., Lactuca sativa L., and Raphanus sativus L.). The addition of HCO 3 ? to a suspension of salt-and HCO 3 ? -epleted thylakoids (suspended in salt-free medium) raised the rate of O2 evolution up to fourfold. This stimulation could be partially replaced by the addition of chloride or nitrate ions. However, the addition of HCO 3 ? in the presence of Cl? or NO 3 ? resulted in a higher stimulation of O2 evolution (sixfold in the presence of nitrate and sevenfold in the presence of chloride). On the other hand, the addition of HCO 3 ? to the thylakoids depleted from salt only raised the rate of O2 evolution by 10–15%, whereas 40–70% was obtained by the addition of nitrate or chloride ions. The fluorescence induction studies indicated a significant decrease in the yield of the variable fluorescence of the salt- and HCO 3 ? -depleted thylakoids. A partial increase in the fluorescence yield was obtained by the addition of HCO 3 ? alone. A typical fluorescence induction curves were obtained by the addition of HCO 3 ? in the presence of Cl? or NO 3 ? ions. The data obtained suggest a similar role for chloride and nitrate ions in O2 evolution in the Hill-reaction, which is restricted at the donor side of photosystem II, whereas bicarbonate plays its role at both sides (acceptor and donor sides). The presented data are those obtained for the thylakoids of P. sativium, which were more or less similar to those obtained for L. sativa and R. sativus.  相似文献   

16.
The effects of carbon, nitrogen, phosphate, and copper on cell growth and production of the isoflavone puerarin by suspension cultures of Pueraria tuberosa (Roxb. ex. Willd.) DC were investigated. Among the various sugars evaluated (glucose, galactose, fructose, maltose, and sucrose), use of sucrose in the medium led to the maximum accumulation of puerarin. A sucrose-feeding strategy in which additional sucrose was added to the flasks 15?d into the culture cycle stimulated both cell biomass and puerarin production. The maximum production of puerarin was obtained when a concentration balance of 20:60?mM NH 4 + /NO 3 ? was used as the nitrogen source. Alteration in the concentration balance of nitrogen components (NH 4 + /NO 3 ? 60:20?mM) or the use of either NH 4 + or NO 3 ? alone decreased biomass production and puerarin accumulation compared with the control culture (NH 4 + /NO 3 ? 20:20?mM). High amounts of phosphate (2.5 and 5?mM) in the medium inhibited puerarin production whereas 0.625?mM phosphate promoted puerarin production (68.3???g/g DW on day?25). An increase in Cu2+ concentration from 0.025 to 0.05?mg/l in the P. tuberosa cell culture medium resulted in a 2.2-fold increase in puerarin production (up to 141???g/g DW on day?25) but reduced cell culture biomass.  相似文献   

17.
High-affinity nitrate transport was examined in intact root hair cells of Arabidopsis thaliana using electrophysiological recordings to characterise the response of the plasma membrane to NO 3 ? challenge and to quantify transport activity. The NO 3 ? -associated membrane current was determined using a three-electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in the roots of seedlings grown in the absence of a nitrogen source, but only 4–6 days postgermination. In 6-day-old seedlings, additions of 5–100 μm NO 3 ? to the bathing medium resulted in membrane depolarizations of 8–43 mV, and membrane voltage (V m) recovered on washing NO 3 ? from the bath. Voltage clamp measurements carried out immediately before and following NO 3 ? additions showed that the NO 3 ? -evoked depolarizations were the consequence of an inward-directed current that appeared across the entire range of accessible voltages (?300 to +50 mV). Both membrane depolarizations and NO 3 ? -evoked currents recorded at the free-running voltage displayed quasi-Michaelian kinetics, with apparent values for Km of 23 ± 6 and 44 ± 11 μm, respectively and, for the current, a maximum of 5.1 ± 0.9 μA cm?2. The NO 3 ? current showed a pronounced voltage sensitivity within the normal physiological range between ?250 and ?100 mV, as could be demonstrated under voltage clamp, and increasing the bathing pH from 6.1 to 7.4–8.0 reduced the current and the associated membrane depolarizations 3- to 8-fold. Analyses showed a well-defined interaction between the kinetic variables of membrane voltage, pHo and [NO 3 ? ]o. At a constant pHo of 6.1, depolarization from ?250 to ?150 mV resulted in an approximate 3-fold reduction in the maximum current but a 10% rise in the apparent affinity for NO 3 ? . By contrast, the same depolarization effected an approximate 20% fall in the Km for transport as a function in [H+]o. These, and additional characteristics of the transport current implicate a carrier cycle in which NO 3 ? binding is kinetically isolated from the rate-limiting step of membrane charge transit, and they indicate a charge-coupling stoichiometry of 2(H+) per NO 3 ? anion transported across the membrane. The results concur with previous studies showing a high-affinity NO 3 ? transport system in Arabidopsis that is inducible following a period of nitrogen-limiting growth, but they underline the importance of voltage as a kinetic factor controlling NO 3 ? transport at the plant plasma membrane.  相似文献   

18.
The effect of nitrate uptake, or its absence, on the utilization of nitrate previously accumulated by dark-grown, decpitated maize (Zea mays L., cv. DeKalb XL-45) seedlings was examined. Five-d-old plants that had been pretreated with 50 mM 14NO 3 ? for 20 h were exposed for 8 h to nutrient solutions containing either no nitrate or 50 mM 15NO 3 ? , 98.7 atom % 15N. The ambient solution, xylem exudate, and plant tissue were analyzed to determine the quantities of previously-accumulated (endogenous) 14NO 3 ? that were translocated to the xylem, lost to the solution, or reduced within the tissue during the 8-h period. Energy was continuously available to the roots from the attached endosperm. In the absence of incoming nitrate, appreciable reduction and translocation of the endogenous 14NO 3 ? occurred, but efflux of 14NO 3 ? to the external solution was minimal. In contrast, during 15NO 3 ? uptake, there was considerable efflux of 14NO 3 ? as well as translocation of 14NO 3 ? to the xylem, but little 14NO 3 ? was reduced. Thus there appeared to be an inverse relationship between 14NO 3 ? efflux and reduction. The data are tentatively interpreted on the basis of a model which envisages (a) two storage locations within roots, one of which primarily supplies nitrate for translocation and the other of which primarily supplies nitrate for outward passage through plasmalemma, and (b) the majority of nitrate reduction as occurring during or immediately following influx across the plasmalemma, with endogenous 14NO 3 ? initially moving outward being recycled inward and thereby being reduced.  相似文献   

19.
The influence of NO 3 ? -N on growth and osmotic adjustment was studied in Tamarix laxa Willd., a halophyte with salt glands on its twigs. Seedlings of T. laxa Willd. were exposed to 1 mM (control) or 300 mM NaCl, with 0.05, 1 or 10 mM NO 3 ? -N for 24 days. The relative growth rate of seedlings at 300 mM NaCl was lower than that of control plants at all NO 3 ? -N levels, but the concentrations of organic N and total N in the twigs did not differ between the two NaCl treatments. Increasing NO 3 ? supply under 300 mM NaCl improved the growth of T. laxa, indicating that NO 3 ? played positive roles in improving salt resistance of the plant. The twigs of T. laxa Willd. accumulated mainly inorganic ions, especially Na+ and Cl?, to lower osmotic potential (Ψs): the contributions of Na+ and Cl? to Ψs were estimated at 31% and 27% respectively, at the highest levels of supply of both NaCl and NO 3 ? -N. The estimated contribution of NO 3 ? -N to Ψs was as high as 20% in the twigs in these conditions, indicating that NO 3 ? was also involved in osmotic adjustment in the twigs. Furthermore, increases in tissue NO 3 ? were accompanied by decreases in tissue Cl? and proline under 300 mM NaCl. The estimated contribution of proline to Ψs declined as with NO 3 ? -N supply increased from 1 to 10 mM, while the contributions of nitrate to Ψs were enhanced under 300 mM NaCl. This suggested that higher accumulation of nitrate in the vacuole alleviated the effects of salinity stress on the plant by balancing the osmotic potential. In conclusion, NO 3 ? -N played both nutritional and osmotic roles in T. laxa Willd. in saline conditions.  相似文献   

20.
Genetic variation at the locus controlling A1 band of erythrocyte esterase was found in the Japanese macaque,Macaca fuscata. Existence of four alleles,Es-A 1 1 ,Es-A 1 2 ,Es-A 1 3 , andEs-A 1 4 , controlling the mobility of the band and codominance relation between them were postulated. A majority of the troops examined were monomorphic inEs-A 1 1-1 phenotype, and the variant phenotypes were observed to occur only in Yugawara-Ihama, Arashiyama, and Koshima areas.  相似文献   

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