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31.
Jesús Arellano Filiberto Vázquez Thelma Villegas Georgina Hernández 《Plant cell reports》1996,15(7):455-458
Summary The sesquiterpene quinone currently known as perezone is abundantly produced by the roots of Perezia cuernavacana. This compound is of biotechnological interest since it may be used as a pigment and has several pharmacological properties. In this work we demonstrate that perezone is also produced in transformed root cultures of P. cuernavacana. Hairy roots were induced by inoculation of internodal segments of sterile plants of P. cuernavacana with Agrobacterium rhizogenes AR12 strain. The axenic liquid MS medium cultures of the hairy roots isolated from the internodes showed active growth in the absence of growth regulators. The transformed nature of the tissue was confirmed by genomic integration (PCR and slot blot hybridization) and expression (enzyme activity) of the marker gus-gene. The production of perezone by a transformed root culture was evidenced by IR spectroscopy. Our results offer an alternative for enhanced production of perezone and represent an advantage over its extraction from natural plant populations which present problems in their agronomic culture. 相似文献
32.
E. Nenz F. Pupilli F. Paolocci F. Damiani C. A. Cenci S. Arcioni 《Plant Cell, Tissue and Organ Culture》1996,45(2):145-152
Culture conditions have been established for callus induction and growth from different explants in L. angustissimus L. Calli were obtained from hypocotyls, leaves, stems, cotyledons and roots cultured on media containing 2,4-dichlorophenoxyacetic acid or -naphthaleneacetic acid with kinetin, N6 – 2 or benzyladenine in different combinations and concentrations. Only those calli induced in presence of -naphthaleneacetic acid with benzyladenine or kinetin produced shoots. Calli induced from hypocotyl explants were the most efficient in regeneration of shoots. Transformation with an Agrobacterium rhizogenes binary vector carrying the plasmid pBI 121.1 is reported. The percentage of cotransformation was estimated by testing GUS activity in hairy roots. The integration of Ri T-DNA and the NPTII gene in transformed plants was confirmed by molecular analyses and in vitro culture of transgenic tissues in the presence of kanamycin.Abbreviations BA
benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- 1AA
indole-3-acetic acid
- NAA
-naphthaleneacetic acid
- 2iP
N6 – 2
- PA
proanthocyanidins
- NOS
nopaline synthase
- NI TII
neomycin phosphotransferase
- GUS
-glucuronidase
- CaMV
cauliflower mosaic virus 相似文献
33.
TransformationofOrychophragmusviolaceusUsingAgrobacteriumtumefaciensAndRegenerationofTransgenicPlantsa¥ZHOUJi-ming(周冀明);WEIZh... 相似文献
34.
L. Martinelli G. Mandolino 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(6-7):621-628
Isolated somatic embryos from petiole-derived callus cultures ofVitis rupestris Scheele have been employed in experiments on genetic transformation. Co-cultivation of somatic embryos during embryogenesis induction withAgrobacterium tumefaciens strain LBA4404, which contains the plasmid pBI121 carrying the neomycin phosphotranspherase and the-glucuronidase genes, produced transformed cellular lines capable of recurrent somatic embryogenesis. Precocious selection for high levels of kanamycin (100 mgl-1) was an important part of our transformation protocol. Transformed lines still have strong-glucuronidase expression as well as stable insertion of the marker genes after 3 years of in-vitro culture, during which they have maintained their capacity to organize secondary embryos and to regenerate transgenic plants with an agreeable efficiency (13%). 相似文献
35.
36.
Nobuyuki Uozumi Yurie Asano Takeshi Kobayashi 《Plant Cell, Tissue and Organ Culture》1994,36(2):183-190
Adventitious shoot primordia were formed on horseradish hairy root cultured in dark. Plantlet formation frequency from the primordia was higher than that from root fragments. Culture for 26 days provided the adventitious shoot primordia, which had the highest potential for plantlet formation (53% explants at 40 days). Benzyladenine supplementation in the dark caused primordium enlargement, but did not increase the number of primordia formed. After adventitious shoot primordia were encapsulated with calcium alginate, kinetin supplementation (2.0–4.0 M) increased the shoot formation frequency (65–80% explants at 20 days) in the light, but also promoted the undesirable formattion of multiple shoots. Supplementation with naphthaleneacetic acid (0.27–5.4 M) in the calcium alginate beads in light enhanced the root emergence from primordia without inhibition of plantlet formation when the encapsulated beads were put on the agar-medium without naphthaleneacetic acid. 相似文献
37.
R. Peñalver B. Vicedo C. I. Salcedo M. M. López 《Biocontrol Science and Technology》1994,4(3):259-267
Agrobacterium radiobacter strains K84, K1026 and K84 Agr‐ produced in vitro an antibiotic‐like substance (ALS 84), different from agrocin 84 and observed in mannitol‐glutamate medium. Twenty five out of 39 A. tumefaciens strains of biovars 1, 2 and 3 were sensitive to ALS 84 regardless of their sensitivity to agrocin 84. Sensitivity in A. tumefaciens strain C58 was not encoded by the Ti‐plasmid. Most isolates tested of Erwinia carotovora subsp. carotovora E. carotovora subsp. atroseptica, Pseudomonas corrugata P. cichorii and unidentified isolates from galls were also sensitive to this substance. ALS 84 was not affected by the proteases studied, nor by treatment at 62°C for 30 min and had a bacteriostatic effect. The production of ALS 84 might play a role in the complex mechanism of biological control of crown gall, especially in strains resistant to agrocin 84 and sensitive to ALS 84, and by the creation of an ecological niche favourable to A. radiobacter strains K84, K1026 or K84 Agr‐. 相似文献
38.
根癌农杆菌对甘蓝型油菜的转化及转基因植株的再生 总被引:37,自引:0,他引:37
用根癌农杆菌(Agrobacterium tum efaciens)共培养法把外源基因导入甘蓝型油菜(Brassi-ca napusL.)主要栽培品种“云北2 号”,获得转基因植株。所用外植体为带有1—2 m m 子叶柄的完整子叶,根癌农杆菌为A208SE(pTiT37-SE, pROA93)。Ti质粒pROA93 带有NPTⅡ及GUS嵌合基因。共培养2 天后转到附加25 m g/L卡那霉素的分化培养基(MS+ 4.5 m g/LBAP)上。AgNO3 和羧苄青霉素促进芽的分化,头孢霉素则有抑制作用。最高转化频率为27% 。把分化出的茎芽切下,插入含有25 m g/L卡那霉素的生根培养基中。羧苄青霉素不利于根的形成。把完整抗性植株移入盛土壤的盆中,生长状况良好。测定β-葡糖苷酸酶活性,84% 明显高于对照。以NPTⅡ基因作探针进行Southern blot分析,证实外源基因已插入到植物细胞基因组中 相似文献
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