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1.
A continuous-flow culture system was developed for culturing Laminaria japonica protoplasts. Protoplasts were settled on 5-μm pore size nylon mesh fixed inside a 50-ml plastic syringe, and cultured in
Provasoli's enriched seawater with iodine medium with a gentle upward flow generated by a peristaltic pump. In the culture
system, 50% of the protoplasts regenerated their cell wall within 24 hours and almost all protoplasts regenerated a cell wall
after 3 days culture. After cell wall regeneration, a number of cells divided and regenerated into sheet-shaped thalli. The
thalli transferred to a tissue culture flask developed into sporophyte-like plantlets within 1 month. Plantlets then differentiated
into blade, stipe, and holdfast, with a proper mucilage canal.
Received: 21 April 1997 / Revision received: 27 June 1997 / Accepted: 5 July 1997 相似文献
2.
An improved procedure for plant regeneration from indica and japonica rice protoplasts 总被引:1,自引:0,他引:1
R. K. Jain G. S. Khehra S. -H. Lee N. W. Blackball R. Marchant M. R. Davey J. B. Power E. C. Cocking S. S. Gosal 《Plant cell reports》1995,14(8):515-519
Plant regeneration from protoplasts of two commercially cultivated Indian indica rice varieties, Pusa Basmati 1 and Java, has been accomplished by plating embryogenic cell suspension-derived protoplasts on the surface of filter membranes overlying agarose-embedded feeder cells of Lolium multltiflorum and Oryza ridleyi, combined with the use of a maltose-containing shoot regeneration medium. Embryogenic cell suspension cultures of Pusa Basmati 1 and Jaya were initiated from mature seed scutellum-derived calli in liquid R2 medium modified by the addition of 560 mg l–1 of proline and 1.0 % (w/v) maltose. In both varieties, protoplast plating efficiencies up to 0.4 % were obtained, depending on the nature of the feeder cells. L. multiflorum feeder cells induced a 6-fold higher plating efficiency than feeder cells of O. ridleyi. In combination, O. ridleyi and L. multiflorum feedercells further enhanced protoplast plating efficiency. Protoplast-derived cell colonies were not obtained from protoplasts of either indica varieties in the absence of feeder cells. MS-based medium containing kinetin (2.0 mg l–1) and -naphthaleneacetic acid (0.5 mg 1–1), together with sucrose and maltose both at 1.5 % (w/v), induced green shoot regeneration in 44 % of protoplast-derived tissues, depending on the feeder cells used for protoplast culture. In both varieties, tissues obtained using O. ridleyi feeder cells were more morphogenic than tissues obtained using L. multiflorum feeder cells, either alone or in combination with cells of O. ridleyi. In the japonica rice variety Taipei 309, this new procedure resulted in a 30-fold increase in plant regeneration from protoplasts compared to previous published procedures.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- GPFs
growth promoting factors
- NAA
-naphthaleneacetic acid
On leave from Department of Genetics, Haryana Agricultural University, Hisar, IndiaOn leave from Biotechnology Centre, Punjab Agricultural University, Ludhiana, India 相似文献
3.
4.
Young embryos of rice (Oryza sativa L. subsp, japonicavar. Guo-xiang No. 1) were cultured on MS agar medium(2,4-D 2 mg/l). Calli were formed and subcultured on N6 agar medium (2,4-D 2 mg/l ). After selection, the small, grainy and pale yellowish cell clusters with dense cytoplasm were used in protoplast preparation. Isolated protoplasts were cultured in N6 medium (2,4-D 1 mg/l, 6-BA 0.2 mg/l)~1 with agarose block culture method. The protoplasts grew, divided and formed calli. After inducing differentiation, the regenerated mature plants were obtained. 相似文献
5.
Kexuan Tang Quanan Hu Enpeng Zhao Aizhong Wu 《In vitro cellular & developmental biology. Plant》2000,36(4):255-259
Summary Factors influencing successful establishment of embryogenic cell-suspension cultures and plant regeneration from longterm
cell suspension-derived protoplasts of the recalcitrant Indica rice cultivar IR36 were studied. The factors included cell
and protoplast culture medium, protoplast culture procedure, the source of nurse cells, and the regeneration procedure. Embryogenic
cell suspension cultures could only be established from mature seed-derived callus of IR36 in AA-based medium (Müller and
Grafe, 1978). Protoplast-derived colonies could be obtained only using the filter-membrane nurse-culture procedure when Lolium multiflorum suspension cells served as nurse, rather than wild rice (Oryza ridleyi) and Japonica rice (Oryza sativa cv. Taipei 309) cells. The utilization of a two-step regeneration procedure led to regeneration of fertile plants from protoplasts
isolated from 2-yr-old cell suspensions of IR36, one of the most important but recalcitrant rice cultivars. 相似文献
6.
7.
目的:建立山茱萸的组织培养及植株再生体系。方法:分别以山茱萸的叶片、花柄和花托为材料,进行山茱萸不同外植体的离体培养研究,筛选最佳培养基组成。结果:适宜山茱萸叶片愈伤组织诱导的培养基组合为1/2MS,附加BA2.0mg/L、IBA0,5—1.0mg/L;适宜山茱萸花柄、花托愈伤组织诱导的培养基组合为1/2MS,附加BA1.0mg/L、2,4-D0.5mg/L;在1/2MS附加BA2.0mg/L、IBA0.05mg/L的培养基上,可诱导不定芽的产生;1/2MS附加IBA2.0mg/L的培养基有利于山茱萸试管苗生根。讨论:山茱萸的花托是进行组织培养的最适外植体,白色或翠绿色、结构致密的愈伤组织较易分化产生不定芽。 相似文献
8.
地黄组织培养及植株再生的研究 总被引:14,自引:2,他引:14
以地黄根茎所获无菌苗为材料,对其愈伤组织诱导、分化和再生植株的获取进行了初步研究。结果表明:取叶片、茎段、叶柄进行愈伤组织诱导,筛选出最适培养基为MS附加2,4-D0.5mg/L、BA1.0mg/L,愈伤组织诱导率可达100%。将叶片接种在分化培养基中,诱导不定芽,其最适分化培养基为MS附加BA 3mg/L、NAA 0.1mg/L,分化率为77.5%。试管苗在改良的MS(大量与微量元素、铁盐和有机物质各1/2)附加NAA 0.05mg/L的培养基上,经过15~20d培养,生根率可达100%。 相似文献
9.
10.
Li Hongchao Machii Hiroaki Hagio Takashi Takezaki Akane Hirabayashi Toshio 《Plant Cell, Tissue and Organ Culture》1999,58(2):119-125
A fast-growing, small, granular, embryogenic callus was selected from primary calli induced from the Japanese wheat cultivar
Nakasoushu and the Australian wheat cultivar Bodallin. Regenerable and fine suspension cultures were induced three to six
months after liquid culture was initiated and were characterized by dense cytoplasm and active division. These suspension
cultures routinely provided high yields of protoplasts with about 90% viability when incubated in a modified KMP (Kao and
Michayluk, 1975) medium containing 1 mg l-1 2,4-D (2,4-dichlorophenoxyacetic acid), and 1 mg l-1 zeatin. Nakasoushu and Bodallin protoplasts divided at frequencies of 8.6% and 11.1%, respectively, in agarose-solidified
media. When Nakasoushu protoplasts were cultured with effective nurse cells of sorghum and wheat, protoplast division increased
to 16.9% and 12.6%, respectively. Plating efficiencies varied from 0.03% to 2.5%. After subculture, protocolonies yielded
embryogenic calli and somatic embryos, from which green plants were eventually regenerated. Whole plants obtained from Nakasoushu
protoplasts were fertile, demonstrating the first report of Japanese cultivars in wheat protoplast cultures.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
11.
Summary In order to study mitogenic control during axolotl limb regeneration, we have developed a primary blastema cell culture as a very sensitive bioassay for blastema mitogens. Transferrin, an iron-binding glycoprotein which has been shown to be the neurotrophic factor for muscle cells, is the mitogen which has been analysed in the present report. Addition of approximately 2 g human transferrin/ ml of serum-free culture medium enhances blastema cell proliferation 11-fold over control levels and 2-fold over that produced by the addition of nerve extracts or purified growth factors extracted from nerve tissues (basic and acidic fetal growth factor, FGF). At a higher concentration (20 g/ml), transferrin alone has no mitogenic effect unless the medium is also supplemented with FeCl3 (100 M). The results are discussed with regard to the sensitivity of the blastema cell culture bioassay and in the context of the neurotrophic theory of urodele limb regeneration. 相似文献
12.
S. Pattnaik C. Pradhan S. K. Naik P. K. Chand 《In vitro cellular & developmental biology. Plant》2000,36(5):407-411
Summary A complete and efficient protocol is presented for plant regeneration from cell-suspension cultures of Dalbergia sissoo Roxb., an economically important leguminous tree. Factors influencing callus initiation, establishment of cell-suspension
culture, callus formation from embredded microcolonies, and shoot organogenesis from suspension-derived callus were identified.
Of the two different auxins tested, callus induction was better on a medium containing naphthalene acetic acid (NAA). The
percentage of callus induction increased considerably when NAA at 2.0 mg l−1 (10.8 μM) was added in conjunction with 0.5 mg l−1 (2.2 μM) N6-benzyladenine (BA). Of the three different explants evaluated for callus induction, hypocotyl segments were most responsive.
Friable hypocotyl-derived callus from the second subculture passage was used to initiate the cell-suspension culture. Optimum
growth of the cell suspension was observed in MS medium supplemented with the same growth regulators as described above for
callus induction, with an initial inoculum cell density of 1%. The plating efficiency of the microcolonies was greatly influenced
by harvesting time and the gelling agent used for plating. Efficiency was highest (93%) with cells harvested at their exponential
growth phase and plated in 1.2 g l−1 Phytagel. Shoot organogenesis from callus cultures was higher on a medium supplemented with a combination of BA and NAA than
on BA alone. Seventy-one per cent of cultures exhibited shoot-bud differentiation on a medium containing 3.0 mg l−1 (13.3 μM) BA and 0.5 mg l−1 (2.7 μM) NAA. Regenerated shoots were rooted on half-strength MS medium containing 1 mg l−1 each of indole-3-acetic acid (5.7 μM), indole-3-butyric acid (4.9 μM) and indole-3-propionic acid (5.3 μM). Plantlets were acclimated and established in soil. 相似文献
13.
骆驼蓬的组织培养及植株再生 总被引:1,自引:0,他引:1
以骆驼蓬(Peganum harmala L)无菌苗下胚轴切段为材料,在不同的培养基上进行愈伤组织的诱导,发现在MS基本培养基附加2.0mg/L 2,4—D、0.5mg/L 6—BA和3%蔗糖时,可100%的诱导出愈伤组织。愈伤组织在附加2.0mg/L 6—BA、0.5mg/L NAA、500mg/L CH和3%蔗糖的MS培养基上诱导出丛生芽,进而发育成苗,苗的分化频率在30%左右。分化苗或其茎切断在附加0.2mg/L IBA、0.2mg/L NAA和3%蔗糖的l/2MS培养基上出现根的分化,分化频率在90%以上。再生植株经炼苗后移栽成活,成活率在80%以上。 相似文献
14.
用4种诱导培养基P1(MS+2,4-D0.5mg/L)、P2(MS+6-BA0.1mg/L+NAA0.5mg/L)、P3(MS+6-BA0.5mg/L+NAA0.5mg/L)、P4(MS+NAA1.0mg/L+KT0.5mg/L),3种分化培养基(MS+6-BA1mg/L;MS+6-BA0.5mg/L;MS+6-BA1mg/L+NAA0.2mg/L)和4种生根培养基(MS;MS+IBA1mg/L;MS+IBA1mg/L+IAA0.5mg/L;1/2MS+IAA0.2mg/L)对苦豆子愈伤组织进行诱导和植株再生,研究影响苦豆子组织培养的因素,结果表明愈伤组织的诱导频率主要依靠激素的种类和浓度,培养基中加入0.2~2mg/L的2,4-D有利于苦豆子愈伤组织生长,但使褐化发生时间提前;培养基中加入活性炭对苦豆子愈伤组织褐化有明显的抑制作用;加入IAA对苦豆子根的分化是必需的。 相似文献
15.
We examined morphologically the process of regeneration before and after fission in a sexual strain of the freshwater planarian
Dugesia japonica. Usually fission takes place in the post-pharyngeal region. Decapitation significantly accelerates the rate
of fissioning. When decapitated worms were treated with substance P and neuropeptide K separately, the rate of fission markedly
decreased in both cases. Before the onset of fission, a presumptive region of fission was recognized in the post-pharyngeal
portion where undifferentiated cells, regenerative cells and newly differentiated cells were localized. Moreover a functional
network of fixed parenchyma cells was noted in this region. After fission, cell distribution in the blastema became quite
different from that of artificially amputated worms. This difference seems to be due to the process that occurs in the presumptive
region of fission.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
16.
A. Jähne P. A. Lazzeri M. Jäger-Gussen H. Lörz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1991,82(1):74-80
Summary We have established embryogenic cell suspension cultures of barley (Hordeum vulgare L. cultivars Igri, Gimpel, Princesse, and Baronesse) from anther-derived embryogenic callus. Suspension cultures of cultivars Igri and Gimpel were regenerable. The most successful cultivar was Igri, from which a number of independent cell lines producing plantlets were established. Plants could be transferred to soil; up to now, 50% of more than 200 regenerated plants were morphologically normal and fertile. The relative frequency of sterile plants increased as suspensions aged. Suspensions older than 1 year produced embryogenic callus but only albino plantlets could be regenerated. 相似文献
17.
In our study, we investigated the effects of regeneration conditions on both green and albino rice plants (Oryza sativa L.). The regeneration frequency of an albino cell line was compared to a normal cell line obtained from mature seed under two kinds of culture conditions; namely, the static culture on semi-solid regeneration medium and the suspension culture in liquid regeneration medium. The albino cell line, from which only albino plantlets were regenerated, was induced from the albino leaf segments. There were no significant differences in the regeneration frequencies between normal and albino calli on the semisolid regeneration medium. On the other hand, the frequency of regeneration of albino calli was significantly lower than that of the control specifically in the liquid regeneration medium. 相似文献
18.
Summary Hexaploid wheat plants were easily regenerated from young embryo-derived callus for twelve genotypes tested. After a 2.5 years culture period, however, most of the callus cells lost their ability to regenerate into shoots, but not into roots.A novel approach was used to regenerate shoots from the long-term suspension cultured cells. In general, instead of selecting embryogenic callus as source material, this approach requires the inoculation of unselected callus into liquid medium followed by removing the free floating cell portion, selecting out non-root forming cell clumps from the root forming primary suspension culture, and growing the putative shoot-competent clumps in liquid medium with reduced auxin concentrations. We have successfully established shoot-competent wheat suspension cultures for cv. Mustang. High (>80%) frequencies of plant regeneration were observed from plating of 2.5 year suspension cultures. The suspension cultures established by this approach have been utilized to select for heat tolerant variants and will be an ideal source material for protoplast culture and transformation studies. This approach can also be applied to other cereal crops which form roots easily but are unstable in maintaining long term regenerable cultures and which are not easily adaptable to suspension culture. 相似文献
19.
James H. Resau Ph.D. John R. Cottrell Kathryn A. Elligett Eric A. Hudson 《Cell biology and toxicology》1987,3(4):441-458
Human esophageal, tracheal, and pancreatic ductal fragments were collected at autopsy after a postmortem interval of 12 hours or less and maintained in explant organ culture for 30 days. The viability and growth of the explants was assessed by morphology, LDH enzyme release, and cellular outgrowth. The viability and growth of the bronchial explant epithelium was directly related to the postmortem interval. Esophageal epithelial regeneration followed the desquamation of the superficial cell layers. Pancreatic epithelia appeared to grow more slowly and with less outgrowth than the other tissues. Epithelial cell growth along the explant surface and onto the culture dish appeared to proceed through the well-characterized process that follows cell injury, i.e., flattening, migration, replication, and differentiation. Thus, sufficient numbers of viable epithelial cells capable of regeneration were present in routine autopsy epithelium, but there was considerable variation from tissue to tissue and case to case. The most effective and accurate approach to follow when evaluating and predicting the growth and viability of these explants is by using a combination of morphologic, enzymatic and biologic assays. Errors in the interpretation of viability are possible when only one assay method is utilized. These tissues grown in explant organ culture are suitable for studies on the mechanism and response of epithelia to cell injury, recovery and wound healing.Abbreviations 4F-1G
4% formaldehyde, 1% glutaraldehyde
- HIFBS
heat inactivated fetal bovine serum
- IA
immediate autopsy
- LDH
lactate dehydrogenase
- OsO4
osmium tetroxide
- RA
routine autopsy 相似文献
20.
Optimization of the conditions for an efficient induction of somatic embryogenic calli and regeneration of plants from mature seeds of japonica rice cultivars was attempted. The number, color, size, shape, and appearance time of the induced embryogenic calli varied among the rice cultivars depending on the type of basal medium (LS, MS, N6). Presence of adequate amount of sucrose in the medium was an absolute requirement for embryogenic callus formation and shoot induction. Induction of the embryogenic calli, whose overall rates ranged from 30 to 56%, was most efficient in N6 medium supplemented with 3.0 mg l–1 of 2,4-D and 30 g l–1 of sucrose. Agar concentration in the regeneration medium was also critical for the shoot induction. Kinetin was found to be more effective for shoot regeneration compared with BA, while the highest shoot regeneration frequencies were observed when either cytokinin was combined with high concentration (2.0 mg l–1) of NAA. The optimal concentration of kinetin for the highest shoot regeneration frequency (6777%) was different among the cultivars tested. The embryogenic calli-derived shoots rooted on a plant growth regulator-free MS medium were successfully established in soil, producing fertile seeds. 相似文献