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1.
Interspecific protoplast fusion between two monokaryotic strains (a methionine auxotrophic and chloramphenicol-resistant Pleurotus ostreatus strain and a wild-type strain of Pleurotus pulmonarius) was carried out to introduce mitochondrial DNA (mtDNA) from P. ostreatus into P. pulmonarius. Because mycelial colonies regenerated on minimum medium containing chloramphenicol only after the treatment of protoplast electrofusion, the regenerants were regarded as protoplast fusants. The fusants isolated from regenerated colonies were uninucleate, and their resistance of chloramphenicol seemed to be a stable trait. The fusants mated with P. pulmonarius but not with P. ostreatus, and showed almost identical random amplified polymorphic DNA (RAPD) patterns to that of the parental P. pulmonarius monokaryon. The sizes of mitochondrial genomes were estimated to be 81.5 kb for P. ostreatus monokaryon, 54.9 kb for P. pulmonarius monokaryon, and 84.5 or 86.0 kb for the fusants. BamHI and EcoRI digest patterns of the fusant mtDNAs were almost the same as the parental P. ostreatus monokaryon. From the above results, the fusants seemed to carry nuclei derived from the monokaryon of P. pulmonarius and mtDNA including the chloramphenicol-resistance gene from the P. ostreatus monokaryon, suggesting that the P. ostreatus mtDNA had been introduced into P. pulmonarius cells by interspecific protoplast fusion. Contribution No. 383 of the Tottori Mycological Institute  相似文献   

2.
In order to develop a novel method of obtaining monokaryons for a mycorrhizal fungus, Lyophyllum shimeji, monokaryotization of dikaryotic stock culture via protoplast formation and regeneration was performed using 12 dikaryotic stocks. From 6 dikaryotic stocks, a total of 120 monokaryons were isolated, and their mating compatibility was tested. Mating-compatible monokaryons were successfully derived from a dikaryotic stock (NBRC 100325), and monokaryons of only 1 mating type relative to the parental dikaryons were isolated from another 3 strains (MH01710, OK2L-1, and HY7L-1). We successfully prepared monokaryotic stocks via protoplast monokaryotization, a technique that can be used to identify biological species of L. shimeji. This technique could be used for breeding various mycorrhizal mushrooms, including Tricholoma matsutake, for which the preparation of monospore cultures is extremely difficult. Part of this study was presented at the 11th Annual Meeting of the Japanese Society of Mushroom Science and Biotechnology (Asahikawa), September 2007  相似文献   

3.
Mycelial protoplasts of two compatible monokaryons ofLentinula edodes were prepared, and electro-fusion was undertaken. Sixteen dikaryons isolated after the electrofusion were tested for their fruiting abilities in comparison to the two dikaryons obtained by reciprocal mycelial mating. There were significant differences in fruiting body yield among the fusants, and some of them produced more fruiting bodies than did normally mated dikaryons. Although these dikaryons were divided into five genetically different groups by isozyme analyses and a cluster analysis, we could not find clear differences among the groups in terms of fruiting body yield, mycelial growth rate, or degree of wood decay. It appeared that fruiting body yield correlated positively with the mycelial growth rate but negatively with the degree of wood decay.  相似文献   

4.
A suitable screening method for heterozygous DNA markers in shiitake,Lentinula edodes (Berk.) Pegler, is reported. Monokaryons were derived from a dikaryon by de-dikaryotization via protoplast formation. Compatibility of the monokaryons was determined by pairwise culture on agar plates. We selected the primers to amplify polymorphic fragments among the original strain (Hokken600H600) and two monokaryons (H600PP-39 and H600PP-67) showing compatibility. A total of 135 fragments were selected as specific random amplified polymorphic DNAs (RAPDs) resulting from 56 primers of the 147 primers tested. Furthermore, we tested whether the polymorphic fragments segregated into 22 among four strains isolated from a basidium. Most of the polymorphic fragments (about 97.8%) showed 22 segregation among the four strains. We concluded that the polymorphic fragments were heterozygous if they were detected in either of the monokaryons (H600PP-39 and H600PP-67) and segregated to 22 among four meiotic strains (H600B-1,-2, -3, and -4). A total of 132 heterozygous DNA markers were therefore selected from a dikaryon of shiitake (Hokken600H600).Part of this report was presented at the 8th Annual Meeting of Mushroom Science and Biotechnology and the 47th Annual Meeting of the Japan Wood Research Society, Kochi, 1997  相似文献   

5.
The purpose of this research was to analyze the karyotype of the interspecific fusants of twoPleurotus species. Auxotrophic mutants derived from the cultivated strain ofP. ostreatus andP. cornucopiae were used. Protoplasts were fused electrically, and the fusants were selected under auxotrophic complementation. Esterase isozyme analysis showed that several fusants had isozyme bands originating from both parental strains, and others had unilateral isozyme bands. The fusant that had expressed isozyme bands of both parental strains showed chromosomal DNA bands of both of the parental strains in pulsedfield gel electrophoresis analysis. Despite the above results, the chromosomal composition of the fusants obtained by the pulsed-field gel electrophoresis did not exhibit all of the bands of both fusion parents.  相似文献   

6.
Colorimetric pH indicators were examined for their use in judging the culture maturity of sawdust-based cultures of shiitake,Lentinula edodes (Berk.) Pegler. Bromophenol blue (BPB) was the preferred indicator for judging culture maturity. Culture pH changes were detected as changes in color from bluish purple to yellow by direct spraying. The pH values of such cultures varied from 6.3 before inoculation to 4.0 during the fruiting stage. Culture maturity assessment can be used to control fruit body flush timing. Measurement of BPB staining inL. edodes sawdustbased cultures showed a good correlation with fruit body yield of the first flush. This method was concluded to be of value for detecting fruiting potential and therefore may be useful for evaluating flush timing forL. edodes cultivators using sawdust-based methods.  相似文献   

7.
Fagaceae species in Japan were identified by restriction fragment length polymorphism (RFLP) and sequence comparison of a region ofrbcL. Of nine restriction endonucleases used for digestion, three (MspI,RsaI,HaeIII) produced different restriction patterns in Fagaceae. Digestion byMspI yielded four patterns: Fagus species,Castanea crenata, Pasania glabra, and others. Digestion byRsaI andHaeIII afforded two patterns:Fagus species and others. These facts indicate thatCastanea crenata andPasania glabra can be identified byMspI restriction patterns ofrbcL. Sequence comparison of a region of therbcL gene among 20 species of Fagaceae showed that: (1) they could be divided into seven groups; (2) there is a site mutation betweenFagus crenata andF. japonica. The latter indicates that the wood of both Fagus species are identifiable at the species level, which is not the case using conventional methods. This result indicates the possibility of wood identification based on DNA polymorphism in Fagaceae at the intrageneric level.Part of this paper was presented at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 3–5, 1996 and the 47th annual meeting of the Japan Wood Research Society, Kochi, April 3–5, 1997  相似文献   

8.
This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at  相似文献   

9.
Focussing on four types of parenchyma cell around pith regions of branches of Robinia pseudoacacia L. var. inermis, we examined the timing and role of cell death during heartwood formation. Large parenchyma cells that were located in the inner part of the pith died within a year. By contrast, other parenchyma cells died within 4 years, with the timing of cell death depending on the type of cell. Axial parenchyma cells of the xylem close to the pith died first. Then, small parenchyma cells died in the perimedullary zone in the outer part of the pith. Finally, ray parenchyma cells in the xylem close to the pith died. Variations in the autofluorescence of cell walls, which might have been due to deposition of heartwood substances, were observed first in xylem ray parenchyma cells and small parenchyma cells in the perimedullary zone. Our results indicate that the initiation of heartwood formation occurs within 4 years in pith regions of branches in Robinia pseudoacacia L. var. inermis. Moreover, it appears that not only xylem ray parenchyma cells but also small parenchyma cells in the perimedullary zone might be involved in the synthesis of heartwood substances.  相似文献   

10.
A method is described for producing viable protoplasts from germinating conidia (germlings) of Ophiostoma picea. The use of MgSO4-based osmotic stabilizers significantly improved protoplast release, as did the use of 20-h-old germlings. Protoplast release rates also increased with higher enzyme concentration and incubation times, but there were associated negative effects, including reduced regeneration. The results indicate that the use of young germlings, low lytic enzyme levels, and short exposure periods produced the most viable protoplasts.  相似文献   

11.
An improved method for selection of large electro-fused protoplasts ofPopulus alba by a micromanipulator was developed. The conditions for electric cell fusion treatment were optimized. For the best result, protoplasts with a cell density of 5 × 105/mL were treated with an alternate current (1 MHz, 200 V/cm) and pulsed with a direct current (2 kV/cm) for 100μs in 2.5 mM CaCl2 and 0.55 M mannitol. The electo-fused protoplasts were cultured in NH4NO3-free Murashige and Skoog’s medium containing 0.6 M of mannitol, 0.09 M sucrose, 1μM of 2,4-dichlorophenoxyacetic acid and 0.1μM of benzyladenine, the same medium used for protoplast culture, but at a very low cell density of 5–10 × 102/mL in a well of a 96-well culture plate. When cell aggregates derived from individual fused protoplasts were transferred to fresh medium with 0, 0.3 or 0.6 M mannitol, large colonies developed. In the shoot differentiation medium, the reaction of the calluses derived from large fused protoplasts towards the growth regulators differed from the non-fused ones. In medium containing 1μM each of naphthalene acetic acid andN-(2-chloro-4-pyridyl)-N′-phenylurea, growth of callus from electro-fused ones was not reduced by much compared to the control, but shoot differentiation was inhibited. Gibberellic acid (0.1–10μM) was beneficial to shoot regeneration; however, irregularly shaped leaves appeared at high gibberellic acid concentrations. Shoots regenerated were rooted in Murashige and Skoog’s medium containing 4μM of indole-3-butyric acid. Some plantlets obtained had a varied morphology. Based on the characteristics of growth, some cells derived from electro-fused protoplasts appear to be physiologically different from the non-fused one.  相似文献   

12.
Y. Abe 《Forest Pathology》1989,19(7):423-434
Hypoxylon truncatum showed itself to occupy a larger area than Lentinus edodes in low-moisture-content stem segments inoculated with both fungi, but the result was just opposite under high moisture conditions. Their mycelial growth and decay rates indicated that H. truncatum is more tolerant of dry circumstances than L. edodes..  相似文献   

13.
The influence of substrate water potential () on the growth and fruiting of three genotypes of shiitake (Lentinula edodes) was investigated. A slight reduction of (–0.5MPa) stimulated mycelial and colony growth on liquid, agar, and sawdust-based substrates.L. edodes has been found to grow well at a around –0.5 MPa, which corresponds to a moisture content around 55%. A small decrease in at the final vegetative growth phase had positive effects on flush quantity. The substrate was significantly affected by the interaction between genotypes and spawn run time. The of well-colonized mature substrate was –0.7MPa before and –4.OMPa after the fruiting. The rose again to –0.7MPa during rapid absorbance of water by soaking, and this rise was repeated during the second and third flushes. It is suggested that the water-holding capacity of a substrate is related to culture maturity. Excellent water-providing capacity (higher) is expected in the substrate of well-matured cultures with a high density of mycelial colonization.Part of this paper was presented at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 1996  相似文献   

14.
Lysiphlebus testaceipes (Cr.), an exotic aphid parasitoid, but widespread along the West Mediterranean and Atlantic coastal areas, was regularly found in the northeastern part of the Iberian Peninsula parasitizing several aphid hosts, inland and in the Pyrenees. It was found that the environmentally induced restriction of available host species together with the opportunistic response and host alternation of the parasitoid contributed to the successful and on-going expansion and establishment of L. testaceipes from the coast to the inland montane areas of the Iberian Peninsula. Also, a comparison of the overall host range of L. testaceipes in several areas of the West Mediterranean/Atlantic (southern France, Spain, Portugal) demonstrated a rapid increase in the host range in these areas.  相似文献   

15.
[目的]探讨miR396在落叶松体细胞胚胎发生中的功能.以期该研究能为解决体胚发育中子叶畸形胚与生根率低的问题提供新的视角,同时也为进一步揭示体胚发育的分子调控网络做基础铺垫.[方法]构建pre-miR396超表达载体,通过农杆菌介导侵染,将pre-miR396转入落叶松胚性细胞中,筛选稳定抗性细胞系.在DNA与RNA水平上,对抗性细胞系进行的鉴定、检测,同时统计比对其与野生细胞系间发芽率、叶片数目以及生根率的差异.[结果]在抗性细胞系基因组中,扩增出HTP与miR396的特异性片段,且无农杆菌Virg基因片段.RNA表达水平上,抗性系pre-miR396与miR396表达皆增加,而靶基因LaGRFs的表达量下调.统计表明抗性系胚的发芽率、生根率、叶片数目畸形率分别为70.60%、0.60%、37%,而野生型分别为92.50%、10.55%、4%,抗性系胚的发芽率、生根率显著降低(Sig.=0.000),叶片数目畸形率显著增加(Sig.=0.000).[结论]miR396在落叶松体细胞胚胎发生过程中,可能通过负调节靶基因LaGRFs的表达或者通过LaGRFs影响与它相互作用的基因,参与体胚子叶原基细胞代谢,从而影响了子叶与叶片的发育;通过负调节LaGRFs或者其它与根部发育相关的靶基因,影响了根端分生区细胞的活动,参与调控根的发育.  相似文献   

16.
Sugar beet, Beta vulgaris L. is a strategic crop of sugar industry in Egypt. It is threatened by several insect pests among most important of them is the beet beetle Cassida vittata. This work deals with the biological control of this insect using four Egyptian and imported entomopathogenic nematodes (EPNs). The nematodes included Steinernema carpocapsae S2, Heterorhabditis bacteriophora S1, S. feltiae and H. bacteriophora (1-3). Daily mortality of larvae, pupae and adults of C. vittata were recorded after treatment with serial concentrations (from 500 to 4,000 infective juveniles/ml) of each of four studied EPNs. Development of nematodes in insect bodies was followed up. S. carpocapsae S2 was chosen for the application against different stages of the pest in a sugar beet field. In the field, single application of S. carpocapsae S2 killed 65% of the larvae, 92% of the pupae and 57.3% of the adults of C. vittata within a week. This work is the first report on using the EPNs to control sugar beet beetle.  相似文献   

17.
The objective of this study was to compare the effects of woodlots of five tree species, continuous maize (Zea mays L.) and natural fallow on soil water and nitrogen dynamics in western Tanzania. The tree species evaluated were Acacia crassicarpa (A. Cunn. ex Benth.), Acacia julifera (Berth.), Acacia leptocarpa (A. Cunn. ex Benth), Leucaena pallida (Britton and Rose), and Senna siamea (Lamarck) Irwin & Barneby). The field experiment was established in November 1996 in a completely randomized block design replicated three times. Maize was intercropped between the trees during the first three years after planting and thereafter the trees were allowed to grow as pure woodlots for another two years. Transpiration by the trees was monitored when they were 3 years old using sap flow gauges. Soil water content was measured using the neutron probe approach between November 1999 and March 2001. Soil inorganic N profiles were measured when the trees were four years old in all treatments. The results indicated that the trees transpired more water than natural fallow vegetation during the dry season. The difference was apparent at a depth of 35 cm soil, but was more pronounced in deeper horizons. The water content in the entire soil profile under woodlots and natural fallow during the dry period was 0.01 to 0.06 cm3 cm−3 lower than in the annual cropped plots. This pattern was reversed after rainfall, when woodlots of A. crassicarpa, A. leptocarpa, A. julifera, S. siamea and L. pallida contained greater quantity of stored water than natural fallow or continuous maize by as much as 0.00 to 0.02, 0.01 to 0.04, 0.01 to 0.04, 0.01 to 0.03 and 0.00 to 0.02 cm3 cm−3, respectively. Natural fallow plots contained the lowest quantity of stored water within the entire profile during this period. Transpiration was greatest in A. crassicarpa and lowest in L. pallida. All tree species examined were `scavengers' of N and retrieved inorganic N from soil horizons up to 2-m depth and increased its concentration close to their trunks. This study has provided evidence in semi-arid environments that woodlots can effectively retrieve subsoil N and store more soil water after rains than natural fallow and bare soil. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

18.
黄竹细胞悬浮培养和原生质体分离*   总被引：8，自引：2，他引：8       下载免费PDF全文
以牡竹属黄竹（ＤｅｎｄｒｏｃａｌａｍｕｓｍｅｍｂｒａｎｃｅｕｓＭｕｎｒｏ）的竹节及无菌苗根颈为外植体，培养于含有５ｍｇ／Ｌ２，４－Ｄ、０．２ｍｇ／ＬＫＴ和２００ｍｇ／ＬＬＨ的ＭＳ培养基上诱导愈伤组织，然后移至相同成份或不同浓度２，４－Ｄ的液体培养基中，进行悬浮培养，以建立分散性良好的悬浮细胞系。利用悬浮细胞和无菌苗叶片经酶解获大量原生质体，其产量分别约为每克鲜重２．５×１０５个和５×１０５个，活力均可达８０％。  相似文献   

19.
The development of Diglyphus isaea (Walker), a parasitoid of leafminers, was studied under laboratory conditions at seven constant temperatures (10, 15, 20, 25, 30, 35, and 40°C) on Liriomyza sativae Blanchard reared on cucumber (Cucumis sativus L.). The total development period (oviposition to adult emergence) decreased with increasing temperature between 15 and 35°C. In 10 and 40°C no development rate was found in D. isaea, so that it may be claimed that these temperatures fall outside the temperature range for development. Linear regression was used to describe the relationship between development rate and temperature. For egg to adult development, males of D. isaea required 153.8 degree-days (DD) above the theoretical threshold of 9.2°C and females required 161.3 DD above 9.4°C. Data were fitted to four nonlinear temperature-dependent models. Evaluation of the models took place based on the following criteria: fit to data, number and biological value of the fitted coefficient, and accuracy on the estimation of the thresholds. It could be concluded that the Briere-1 and Briere-2 models are suitable for estimating the minimum, maximum and optimal temperature thresholds of D. isaea. Thermal requirements and temperature thresholds can be used to predict the occurrence, number of generations and population dynamics of D. isaea.  相似文献   

20.
To find fungi that are potent for degradation of condensed tannin, a two-step screening was used. This involved measurement of fungal growth rate on Japanese cedar (Cryptomeria japonica) bark, followed by determination of [14C]-labeled CO2 generated from fungal degradation of synthetic [14C]-labeled condensed tannin model. In the first screening, 75 strains of wood rot fungi were tested, and 19 strains effectively decreased bark weight and/or the weight of the methanol-soluble fraction. For the second screening, [14C]-labeled condensed tannin model compound was synthesized in 11.8% yield based on radioactivity measurements. Over the incubation period, Coriolus hirsutus K-2671, Lentinus edodes Is, and Lampteromyces japonicus Nn showed higher cumulative [14C]-labeled CO2 emissions than the other strains and mineralized the [14C]-labeled condensed tannin model compound by 3.7%, 3.0%, and 3.0%, respectively. Fractionation of the methanol extracts from the medium by gel permeation chromatography after fungal treatment suggested that fungi that can induce the emission of significant levels of [14C]-labeled CO2 can extensively depolymerize condensed tannins.  相似文献   

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