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1.
Summary Seventeen cultivars belonging to the genus Dianthus were examined for protoplast isolation, culture and shoot regeneration under the same conditions. These included D. caryophyllus, D. chinensis, D. barbatus, D. plumarius, D. superbus and D. japonicus as well as interspecific hybrid cultivars (D. caryophyllus x D. chinensis and D. chinensis x D. barbatus). In all cultivars, viable protoplasts were isolated at high yields from leaves of axenic shoot cultures and some of these protoplasts divided and formed colonies. However, shoot regeneration frequencies were markedly different among the species. High frequency shoot regeneration was obtained from D. chinensis and interspecific hybrid cultivars, while only low frequency or no shoot regeneration was obtained from other species.Abbreviations MS
Murashige and Skoog (1962)
- FW
fresh weight
- MES
2-N-morpholinoethane sulfonic acid
- FDA
fluoroscein diacetate
- NAA
1-naphthaleneacetic acid 相似文献
2.
Sumiyuki Mii Jennifer Duong Yasunori Tome Aisada Uchugonova Fang Liu Yasuyuki Amoh Norimitsu Saito Kensei Katsuoka Robert M. Hoffman PhD 《Journal of cellular biochemistry》2013,114(7):1674-1684
We have previously reported that nestin‐expressing hair follicle stem cells can differentiate into neurons, Schwann cells, and other cell types. In the present study, vibrissa hair follicles, including their sensory nerve stump, were excised from transgenic mice in which the nestin promoter drives green fluorescent protein (ND‐GFP mice), and were placed in 3D histoculture supported by Gelfoam®. β‐III tubulin‐positive fibers, consisting of ND‐GFP‐expressing cells, extended up to 500 µm from the whisker nerve stump in histoculture. The growing fibers had growth cones on their tips expressing F‐actin. These findings indicate that β‐III tubulin‐positive fibers elongating from the whisker follicle sensory nerve stump were growing axons. The growing whisker sensory nerve was highly enriched in ND‐GFP cells which appeared to play a major role in its elongation and interaction with other nerves in 3D culture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. The results of the present report suggest a major function of the nestin‐expressing stem cells in the hair follicle is for growth of the follicle sensory nerve. J. Cell. Biochem. 114: 1674–1684, 2013. © 2013 Wiley Periodicals, Inc. 相似文献
3.
Hairy roots of snapdragon (Antirrhinum ma-jus L.: Scrophulariaceae) induced by a wild-type strain of Agrobacterium rhizogenes were cultured on media containing various concentrations of a phosphinothricin-based herbicide, bialaphos, or plant growth
regulators (PGRs). Adventitious shoot regeneration from hairy roots was observed with a low frequency (10%) on half-strength
Murashige and Skoog medium. Addition of α-naphthalene-acetic acid in combination with 6-benzylaminopurine, thidiazuron, or zeatin to the medium had no effect on shoot
regeneration from hairy roots. Although bialaphos at 0.9 mg l–1 or more was toxic to hairy roots, it significantly increased the shoot regeneration frequency up to 56% at 0.5 mg l–1. In contrast, non-transformed roots and leaves regenerated no shoots on media with or without bialaphos. Regenerated shoots
detached from host roots readily developed roots on gellan-gum-solidified medium. Regenerated plants were successfully transferred
to the greenhouse, but did not produce seed.
Received: 24 February 1997 / Revision received: 10 July 1997 / Accepted: 28 July 1997 相似文献
4.
Genetic transformation was carried out with wild-type strains of Agrobacterium rhizogenes for introducing a dwarf trait into the Scrophulariaceous ornamental plant, angelonia (Angelonia salicariifolia). Leaf segments of two angelonia genotypes (Ang.1 and Ang.2) were co-cultivated with mikimopine-type strains of A. rhizogenes. Adventitious roots that showed vigorous growth and increased lateral branching when cultured on half-strength Murashige and Skoog's (MS) basal salts medium lacking plant growth regulators (PGRs) after co-cultivation were selected as putatively transformed lines. All of these selected lines produced mikimopine. Adventitious shoots were efficiently induced from putatively transformed root segments on half-strength MS basal salts medium containing 1 mg l(-1) benzyladenine (BA) under continuous illumination (24-h photoperiod), and the shoots easily rooted following their transfer to half-strength MS basal salts medium lacking PGRs. The transgenic nature of regenerated plants was confirmed by Southern hybridization. Transformed plants frequently died during their acclimatization, and acclimatized plants of eight transformed lines grew very slowly for 1-5 months after transplantation to the greenhouse. Plants of two transformed lines of Ang.2 flowered 4-6 months after transplantation. These transformed plants exhibited phenotypic alterations such as dwarfness and smaller leaves. There were no apparent alterations observed in the number, shape, and size of the flowers. Pollen fertility of the transformed plants was 60-80% based on aceto-carmine staining. These results indicate the possibility of applying A. rhizogenes-mediated transformation for introducing a dwarf trait into angelonia. 相似文献
5.
Plant regeneration from protoplasts of Lilium × formolongi cv. Azusa was achieved by utilizing suspension cultures of meristematic nodular cell clumps with a high plant regeneration ability. Creamy-white calli with embryogenic potential were initially induced from the seeds on Murashige and Skoog (MS) medium containing 4.1 μM picloram. The calli were then transferred into a liquid medium of the same composition, in which they turned into compact cell clumps which consisted of meristematic nodules. Protoplasts were readily isolated from these meristematic nodular cell clumps. Colonies were successfully formed from the protoplasts by embedding in 0.1% gellan gum-solidified MS medium containing 4.1 μM picloram and 0.5 M glucose. They regenerated shoots and roots on MS medium containing 2.2 μM benzylaminopurine (BAP). The plants thus obtained produced flowers with normal fertile pollen 8 months after successful transfer into soil. These plants had normal chromosome numbers (2n = 24) but had shorter leaves than original plants. They set seeds after as well as cross pollination. 相似文献
6.
Protoplasts isolated from cell suspension culture of Phalaenopsis “Wataboushi” were cultured by (a) embedding in gellan gum-solidified hormone-free 1/2 New Dogashima medium (1/2 NDM) containing
0.44 M sorbitol, 0.06 M sucrose and 0.1 g/l l-glutamine (standard method) and (b) beads method using beads of gellan gum or sodium alginate as the gelling agents which
were surrounded by liquid NDM. Although, the two beads methods gave less frequency of initial protoplast division than the
standard method, the former finally resulted in higher frequency of microcolony formation than the latter. The highest frequency
of microcolony formation (23%) was obtained when protoplasts were embedded in 1% Ca-alginate beads and subcultured every two
weeks by replacing the surrounding liquid culture medium with a decrease in sorbitol concentration by 0.1 M. Colonies visible
to the naked eyes were observed within 2 months of culture and the regenerated calluses were transferred onto hormone-free
NDM supplemented with 10 g/l maltose and 0.3% (w/v) gellan gum, on which PLBs were formed and proliferated profusely. The
PLBs were regenerated into plantlets after changing the carbon source to 10 g/l sorbitol and successfully acclimatized to
greenhouse conditions. 相似文献
7.
Raham Sher Khan Gunaratnam Thirukkumaran Ikuo Nakamura Masahiro Mii 《Plant Cell, Tissue and Organ Culture》2010,101(3):279-285
The presence of marker genes conferring antibiotic or herbicide resistance in transgenic plants has been a serious problem
for their public acceptance and commercialization. MAT (multi-auto-transformation) vector system has been one of the strategies
to excise the selection marker gene from transgenic plants. Agrobacterium
tumefaciens strain EHA105 harboring a rol-type MAT vector, pMAT101, was used to produce morphologically normal transgenic Petunia hybrida ‘Dainty Lady’ employing rol gene as the selection marker gene. LacZ gene was used as a model gene of interest. Infected explants were cultured on plant growth regulator (PGR)- and antibiotic-free
half-strength MS medium. Sixty-five percent of the infected explants produced hairy roots. The hairy roots were separated
and proliferated on 1/2 MS hormone-free medium. Shoots produced from the hairy roots on 1/2 MS medium supplemented with benzylaminopurine
(BA) and naphthalene acetic acid (NAA) exhibited hairy root syndrome (Ri syndrome) such as dwarfed, reduced apical dominance,
short internodes and increased rooting, but subsequently produced normal-looking marker-free shoots. Molecular analysis of
DNA from the hairy roots, shoots with Ri syndrome and morphologically normal shoots revealed that the normal shoots had only
LacZ gene, and the removable cassette consisting of rol, R (recombinase) and GUS genes was excised. From this study it can be concluded that the chimeric rol genes can be used as a selection marker for Agrobacterinum-mediated transformation of Petunia hybrida and that the production of marker-free normal transgenic plants is possible without using selective chemical agents employing
rol-type MAT vector. 相似文献
8.
Supaporn Hasthanasombut Valentine Ntui Kanyaratt Supaibulwatana Masahiro Mii Ikuo Nakamura 《Plant biotechnology reports》2010,4(1):75-83
Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants.
We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In
this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants
expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of T2 progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results
indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated
normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest
that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance. 相似文献
9.
Fumihiro Fujiki Yoshihiro Oka Mai Kawakatsu Akihiro Tsuboi Hiroko Nakajima Olga A. Elisseeva Yukie Harada Zheyu Li Naoko Tatsumi Eriko Kamino Toshiaki Shirakata Sumiyuki Nishida Yuki Taniguchi Ichiro Kawase Yusuke Oji Haruo Sugiyama 《Microbiology and immunology》2008,52(12):591-600
The Wilms' tumor gene WT1 is overexpressed in various tumors, and the WT1 protein has been demonstrated to be an attractive target antigen for cancer immunotherapy. A WT1 protein‐derived 16‐mer peptide, WT1332 (KRYFKLSHLQMHSRKH), which was naturally generated through processing in cells and could elicit Th1‐type CD4+ helper T cell responses with an HLA‐DRB1*0405‐restriction has previously been identified by us. In the present study, it has been demonstrated that WT1332 can induce WT1332‐specific CD4+ T cell responses with the restriction of not only HLA‐DRB1*0405 but also HLA‐DRB1*1501, ‐DRB1*1502, or ‐DPB1*0901. These HLA class II‐restricted WT1332‐specific CD4+ T cell lines produced IFN‐γ but neither IL‐4 nor IL‐10 with WT1332 stimulation, thus showing a Th1‐type cytokine profile. Furthermore, HLA‐DRB1*1501 or ‐DRB1*1502‐restricted WT1332‐specific CD4+ T cell lines responded to WT1‐expressing transformed cells in an HLA‐DRB1‐restricted manner, which is consistent with our previous finding that WT1332 is a naturally processed peptide. These results indicate that the natural peptide, WT1332, is a promiscuous WT1‐specific helper epitope. WT1332 is expected to apply to cancer patients with various types of HLA class II as a WT1‐specific helper peptide in combination with HLA class I‐restricted WT1 peptides. 相似文献
10.
Evaluation of 12 β-lactam antibiotics for Agrobacterium-mediated transformation through in planta antibacterial activities and phytotoxicities 总被引:2,自引:0,他引:2
The antibacterial activities of 12 beta-lactam antibiotics against Agrobacterium tumefaciens strains LBA4404 and EHA101 living in tobacco (Nicotiana tabacum L.) leaf tissues, and their phytotoxicities to tobacco leaf tissues were evaluated. All beta-lactams at minimum bactericidal concentration (MBC) or higher showed weak bactericidal activities against agrobacteria persisting in tobacco leaf tissues. The beta-lactams evaluated were classified into two major groups according to their inhibitory effect on shoot regeneration of tobacco leaf tissues: (1) highly phytotoxic drugs, and (2) moderately phytotoxic drugs. According to these results, suitable kind and concentration of beta-lactam antibiotics were evaluated for Agrobacterium-mediated transformation. 相似文献