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1.
Regeneration of white spruce (Picea glauca) somatic embryos from protoplasts derived from an embryogenic suspension culture was accomplished using a culture medium containing 2 mgl–1 2,4-D and 1 mgl–1 6-BAP. Divisions within 2 days led to plating efficiencies in the order of 24% after 9 days. A reduction in the osmoticum, necessary for sustained growth, was carried out gradually over 30 days. Embedding in agarose and culture in 5 cm petri dishes prior to transfer of agarose blocks to a bead type culture, led to the formation of somatic embryos as early as 23 days after isolation and yielded plating efficiencies in the order of 5–10% after 35 days culture. 相似文献
2.
Fifteen pinealectomized and 15 unoperated ewes were exposed to constant light for 3 weeks before and 10 weeks after lambing. Fourteen pinealectomized and 15 unoperated ewes were allowed to lamb outdoors. Five ewe lambs born in constant light to the 2 groups of dams were pinealectomized at 10 weeks of age. Ewes and lambs were then returned to the field. Puberty (determined by weekly progesterone analysis) was significantly delayed (P less than 0.05) in the pinealectomized ewe lambs. Median pubertal age in pineal-intact ewe lambs was 37 weeks compared to 49 weeks in pinealectomized lambs. Constant light during the first 10 weeks of life had no effect upon puberty onset nor did the pineal status of the dam. Control lambs entered seasonal anoestrus at the time pinealectomized ewe lambs were entering puberty. Pinealectomized lambs entered anoestrus at the same time as control lambs were beginning their second breeding season. These results confirm a key role of pineal-mediated hormonal signals in the control of puberty in the sheep. 相似文献
3.
David I. Dunstan Keith E. Turner Wayne R. Lazaroff 《Plant Cell, Tissue and Organ Culture》1985,4(1):55-60
The quality of shoots in cultures of the apple rootstock, M4, was used as a criterion for the selection of an optimum medium.
The frequency of shoots in defined shoot clases was monitored for each of five media, which differed in the type and concentration
of phytohormone. Media containing BA (1.15 mg l-1) and IBA (either 0.15 or 0.20 mg l-1) produced the maximum number of shoots that were desirable for transplantation and acclimatization. 相似文献
4.
F. Bekkaoui R. S. S. Datla M. Pilon T. E. Tautorus W. L. Crosby D. I. Dunstan 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,79(3):353-359
Summary Protoplasts from suspension cultures of somatic embryos of white spruce (Picea glauca Moench Voss) were electroporated with plasmids containing the chimeric genes for chloramphenicol acetyl transferase (CAT) or -glucuronidase (GUS), under control of one of three promoters. Transient CAT gene expression of approximately equal magnitude resulted when the CAT gene was fused to either the cauliflower mosaic virus (CaMV) 35S promoter or the nopaline synthase (NOS) promoter. When the CAT gene was fused to a tandem repeat CaMV 35S promoter (pPBI-363), CAT enzyme activity compared to NOS or 35S promoters increased up to eightfold (cell line WS-34), and were up to 100-fold greater than control (electroporated without plasmid). Comparatively, protoplasts of black spruce (Picea mariana Mill) and jack pine (Pinus banksiana Lamb.), electroporated with pPBI-363, produced increases in CAT activity compared to control of 90-fold and 70-fold, respectively. White spruce (WS-34) protoplasts were subsequently electroporated with the GUS gene fused to the tandem repeat CaMV 35S promoter. Comparatively, GUS enzyme activity increased up to tenfold compared to GUS fused to a CaMV 35S promoter. The results indicated that transient expression of the CAT and GUS genes was influenced by the type of promoter and cell line used, as well as by electroporation conditions.NRCC No. 30498 相似文献
5.
6.
T. E. Tautorus M. M. Lulsdorf S. I. Kikcio D. I. Dunstan 《In vitro cellular & developmental biology. Plant》1994,30(1):58-63
Summary Embryogenic cell cultures ofPicea mariana (black spruce) and the species complexPicea glauca-engelmannii (interior spruce) were maintained either as suspensions in liquid medium in 250 or 500-ml-capacity shake-flasks, 7-liter-capacity
airlift or mechanically stirred bioreactors, or on agar-solidified medium. Cultures from each of the maintenance conditions
were subsequently transferred to agar-solidified LP medium containing 40μM (±) -abscisic acid for maturation into cotyledonary stage embryos. For both species, the highest maturation frequency resulted
from cultures grown in the airlift bioreactor. With black spruce cells grown in the airlift bioreactor containing LP medium
with 60 mM sucrose, a maximum of 7.1 g·liter−1 dry weight and 2892 embryos·ml−1 were obtained after 15 days. For interior spruce cells, a maximum dry weight of 5.9 g·liter−1 and 2698 embryos·ml−1 were obtained after 21 to 30 days. During culture over 2 wk, ammonia was almost completely utilized by both species, wherease
nitrate was depleted to 40% of the initial concentration. Sucrose was rapidly hydrolyzed to glucose and fructose by both species.
Black spruce cultures preferentially metabolized glucose, whereas interior spruce preferentially metabolized fructose. Improved
growth of interior spruce cells in mechanically stirred bioreactors occurred when cultured in LP medium with 60 mM fructose as the sole carbon source.
NRCC no. 36479 相似文献
7.
David I. Dunstan Terry D. Bethune Cheryl A. Bock 《In vitro cellular & developmental biology. Plant》1993,29(3):109-112
Summary The production of cotyledonary somatic embryos of white spruce from cultures grown long-term as suspensions was investigated.
We report the effects of removal of 2,4-dichlorophenoxyacetic acid (2,4-D) from the maintenance medium (ordinarily containing
both 2,4-D and benzyl adenine) before (±)-ABA-stimulated maturation. In particular the use of a 1-wk culture period
without 2,4-D was found to improve the production of normal-looking cotyledonary somatic embryos. Using high performance liquid
chromatography analyses of culture supernatants, it was determined that this affect was not related to altered ABA metabolism.
Germination of cotyledonary somatic embryos from cultures pretreated by the 1-wk culture period without 2,4-D was improved
compared with similar embryos from cultures that had not been pretreated. 相似文献
8.
Molecular characterization of the staphylococcal multidrug resistance export protein QacC. 总被引:10,自引:2,他引:8 下载免费PDF全文
The QacC polypeptide is a member of a family of small membrane proteins which confer resistance to toxic compounds. The staphylococcal qacC gene confers resistance to toxic organic cations via proton-dependent export. The membrane topology of the QacC polypeptide was investigated by constructing and analyzing a series of qacC-phoA and qacC-lacZ fusions. From these analyses, most of the predicted features of the QacC protein were verified, although data regarding the possible orientation of the COOH region were not conclusive. The role of the sole cysteine residue, Cys-42, in QacC was studied by using the sulfhydryl reagent N-ethylmaleimide and site-directed mutagenesis. N-Ethylmaleimide was shown to inhibit qacC-mediated ethidium export. Multiple amino acid substitutions were made for Cys-42, and mutations at this location had various effects on resistance specificity. This suggests that the Cys-42 residue may be located near a region of QacC that is involved in substrate recognition. Mutagenesis of conserved residues in QacC indicated that Tyr-59 and Trp-62 also play an essential structural or functional role in QacC. 相似文献
9.
Summary Cells of the scutellum of immature embryos ofSorghum bicolor plated onto an agar medium containing 2,4-D give rise to shoots and embryo-like structures and to some callus. Some of the embryo-like structures later develop into typical sorghum embryos complete with scutellum, coleoptile and coleorhiza. The results of anatomical studies of the development of these secondary growth forms by light and scanning electron microscopy suggest that shoots and embryo-like structures can arise directly from cells of the primary scutellum without an intervening callus phase. In some cases it appears that the scutellum of the secondary embryos arises by folding of the scutellum of the sexual embryo and does not arisede novo. In other cases the structures arise from single cells. No evidence was found to indicate that organized structures arose from proliferating callus cells. The unorganized callus which arises initially is not capable of growth through continuous subculture; it produces a purple-black pigment and rapidly becomes necrotic. The significance of these observations is discussed in relation to present views on morphogenesis in cereal cell cultures and their implications forin vitro cell genetics.Abbreviations used in the text 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog 相似文献
10.
David I. Dunstan Debbie P. Lashta Susan I. Kikcio Trevor A. Thorpe 《In vitro cellular & developmental biology. Plant》1992,28(1):33-38
Summary The effects of sucrose concentration, addition of ammonium nitrate, and exposure to N6-benzyl-adenine (BA) on multiplication potential with shoots derived from shoot cultures of 17- to 20-yr-old Douglas fir trees
[Pseudotsuga menziesii (Mirb.) Franco] were compared. Each of these conditions, when compared independently, affected recurrent shoot multiplication
and influenced shoot development, as measured by the abundance of shoot apices. Sucrose concentration was influential, the
use of 25 g · liter−1 providing twice the multiplication obtained with 20 g · liter−1, and 14 × that obtained with the 30 g · liter−1 concentration routinely used (tree 11). Ammonium nitrate usage also improved multiplication, a 2.5 times improvement being
obtained after incorporation of 100 mg · liter−1 NH4NO3 into the medium (tree 33). Shoot cultures were responsive but relatively sensitive to addition of BA, the best improvement
in multiplication (5 times) being obtained with brief exposures to 3 mg · liter−1 BA (tree 11). Although shoot cultures were responsive to the conditions investigated, differences in shoot multiplication
and development were not displayed for several weeks. It was not possible therefore to repeat all the treatments with more
than one genotype; however, when this was possible a genotype-dependent variation in response was evident. 相似文献