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11.
单盐(KCl, CaCl_2或MgCl_2)和混合盐(KC_1+CaCl_2或KCl+MgCl_2)对植物原生质体完整率、存活率和膜透性等均有明显影响。K~+、Ca~(2+)或Mg~(2+)等单种阳离子明显降低原生质体膜完整率和存活率而增加其物质渗漏量,其中以单价阳离子K~+的影响为甚。上述单种阳离子还明显降低小麦幼叶超氧物歧化酶(SOD)和过氧化氢酶活性。只有由单价和二价阳离子组成的平衡混合盐才能使原生质体维持较高的完整率、存活率和较正常的膜透性.并能使细胞维持较高的SOD和过氧化氢酶活性。 认为单盐毒害机理可能是首先引起细胞膜发生不正常的膜相变或细胞累积较多的有害氧自由基,引起膜脂发生过氧化或脱酯化而破坏膜结构。在离子平衡混合盐作用下,膜系才能维持正常液晶相,具有较高活性的SOD和过氧化氢酶等生物保护性酶系是离子拮抗作用之原因。 相似文献
12.
L. Tanno-Suenaga H. Ichikawa J. Imamura 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1988,76(6):855-860
Summary X-irradiated protoplasts of Daucus carota L., 28A1, carrying cytoplasmic male sterile (CMS) cytoplasm and iodoacetamide-treated protoplasts of a fertile carrot cultivar, K5, were fused with polyethylene glycol (PEG), and 73 plants were regenerated. Twenty-six randomly chosen regenerated plants had non-parental mitochondrial DNA (mtDNA) as revealed by XbaI restriction fragment patterns, and all of the plants investigated had diploid chromosome numbers. Of the 11 cybrid plants that showed mtDNA fragment patterns clearly different from those of the parents, 10 plants showed male sterility with brown or red anthers, and one plant possessed partially sterile yellow anthers. The mtDNA fragment patterns of the ten cybrid plants with male sterile flowers resembled that of a CMS parent, 28A1; and four fragments were identified that were common between the sterile cybrid plants and 28A1, but absent from the partially sterile cybrid plants and a fertile cultivar, K5. The results indicated that the CMS trait of the donor was efficiently transferred into the cybrid plants by donor-recipient protoplast fusion. 相似文献
13.
Kirsten Brandt 《Plant Cell, Tissue and Organ Culture》1991,26(3):195-201
A method for estimating the oxygen availability in plant cell cultures grown in stationary liquid media (e.g. many protoplast cultures) was developed. The method is based on short-term measurements of respiration rate versus oxygen concentration on a sample of cells, suspended in liquid media. From such data it is possible to estimate the oxygen concentration at the bottom of a stagnant liquid culture, by calculating the amount of oxygen reaching the cells by diffusion. As an example, rape (Brassica napus L. cv. Omega) hypocotyl protoplasts were grown with different oxygen concentrations at the site of the cells, obtained by varying the cell density, the height of the liquid layer and the oxygen content of the gas phase. The number of surviving calli was positively correlated with the estimated oxygen availability in the range between 60 and 350 M O2, below 60 M all cells died. This indicates that oxygen availability can be a limiting factor in the range usually encountered in protoplast cultures, and that the method can be useful when designing optimal growth conditions for stationary cultures of plant cells.Abbreviations C1
bulk oxygen concentration in agitated medium
- Co
oxygen concentration in medium at the gas-liquid interface, in equilibrium with the gas
- Cx
oxygen concentration at cell level
- D
diffusion constant of oxygen in water
- KLa
oxygen transfer rate
- l
height of liquid above cells
- n
number of cells per ml
- Rx
respiration rate per cell 相似文献
14.
A protoplast to plant system in roses 总被引:7,自引:0,他引:7
Derek Matthews John Mottley Imelda Horan Andrew V. Roberts 《Plant Cell, Tissue and Organ Culture》1991,24(3):173-180
High yields of protoplasts were isolated from embryogenic suspension cultures of Rosa persica x xanthina and Rosa wichuraiana using an enzyme mixture comprising 20 g l-1 cellulase Onozuka R10, 1 g l-1 Pectolyase Y-23 and 10 g l-1 hemicellulase. Agarose-immobilized protoplasts gave the most consistent growth at a plating density of 5×104 protoplasts ml-1 on the basic medium of Kao & Michayluk (KM8p) containing 2 mg l-1 naphthaleneacetic acid and 1 mg l-1 benzylaminopurine. At 25°C in the dark, 0.004% of R. persica x xanthina protoplasts developed into colonies. Using similar culture conditions, but with a plating density of 9×104 protoplasts ml-1, 0.017% of R. wichuraiana protoplasts developed into colonies. On transfer of R. persica x xanthina colonies to Schenk & Hildebrandt's medium containing 3 mg l-1 2,4-dichlorophenoxyacetic acid, globular and later stage embryos were formed. Approximately 30% of these embryos developed into plantlets on transfer to basal Schenk & Hildebrandt's medium. Further development of the plantlets took place on cellulose plugs (Sorbarods) soaked in Murashige & Skoog's medium containing 0.05 mg l-1 naphthaleneacetic acid, 0.05 mg l-1 indole-3-butyric acid and 0.1 mg l-1 benzylaminopurine. Rose breeding is now open to the full range of in vitro genetic manipulation techniques involving protoplast technology. 相似文献
15.
M. Dijak A. Sproule W. Keller R. Pandeya S. Gleddie 《Plant Cell, Tissue and Organ Culture》1991,25(3):189-197
Agrobacterium tumefaciens strains harbouring plasmid vectors pBCAT1, pVU1011 or pMON806 were used to transform leaf explants of Nicotiana tabacum cultivars Delgold and Candel, N. debneyi, and N. rustica var. NRT. Transgenic plants resistant to the selective agents kanamycin, hygromycin or methotrexate were regenerated and used as sources of leaf mesophyll protoplasts. Protoplasts divided and regenerated plants in the presence of selective agents at levels inhibitory to protoplasts of non-transformed plants. Cross-resistance of protoplasts to more than one selective agent was not observed in this study which suggests that this approach may lead to an efficient interspecific somatic hybrid selection system. 相似文献
16.
S. J. Ochatt 《Plant Cell, Tissue and Organ Culture》1991,25(2):161-167
Leaf protoplasts of axenic shoot cultures of Lonicera nitida cv Maigrun underwent sustained division to give multicellular colonies (microcalli) on a modified, ammonium-free MS (Murashige & Skoog) medium containing 0.5 mg l-1 NAA (1-naphthaleneacetic acid), 1.0 mg l-1 BAP (6-benzylaminopurine) and 150 mg l-1 casein enzymatic hydrolysate. Callus was produced upon transfer of cell colonies to MS medium with 2.0 mg l-1 NAA and 0.2 mg l-1 BAP. About 110 days from isolation protoplast-derived shoots were regenerated on a half-strength MS medium with 0.01 mg l-1 NAA, 5.0 mg l-1 BAP, 0.5 mg l-1 zeatin and a complex mixture of group B vitamins. The replacement of such mixture by 250 mg l-1 casein enzymatic hydrolysate promoted rhizogenesis in calli, with shoot buds being subsequently regenerated from the protoplast-derived roots. Micropropagation of protoplast-derived shoots (of either origin) was difficult, due to a strong apical dominance, but could be accomplished by transferring single-node explants to half-strength MS medium with 1.5 mg l-1 BAP. Such shoots were, in turn, successfully rooted and transferred to the glasshouse where they completed acclimatization.Abbreviations BAP
6-benzylaminopurine
- CPW
Power et al. (1989) medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- F.P.E.
final plating efficiency
- f.wt.
fresh weight
- IAA
4-indole-3yl-acetic acid
- IBA
4-indole-3yl-butyric acid
- I.P.E.
initial plating efficiency
- MES
2-N-morpholinoethane sulfonic acid
- M.P.E.
intermediate plating efficiency
- MS
Murashige & Skoog (1962) medium
- NAA
1-naphthaleneacetic acid
- PVP-10
polyvinylpirrolidone
- Av MW 10,000, TIBA
2,3,5-tri-iodobenzoic acid
- Z
zeatin 相似文献
17.
J. Ballongue J. Amine E. Masion H. Petitdemange R. Gay 《FEMS microbiology letters》1985,29(3):273-277
Abstract Factors that may initiate the biosynthesis of acetoacetate decarboxylase were investigated in resting cells of Clostridium acetobutylicum . Linear acids from C1 to C4 were inducers, whereas branched acids and linear acids from C5 to C7 were not inducers of acetoacetate decarboxylase biosynthesis. Induction of acetoacetate decarboxylase was maximal at pH 4.8 in the presence of acid concentrations comparable with those found during fermentation. In growth conditions repression of acetoacetate decarboxylase biosynthesis was found. This fact explains that acetone production by Clostridium acetobutylicum occurs when growth slows down. 相似文献
18.
应用一交变电场(正弦波,500KHz,175—225V/cm),分别将裸大麦(正常或正常与黄化)、蚕豆、烟草的叶肉原生质体在一定间隔、平行的两电极间排列成串。再附加单个的方波脉冲(10—40μs,800V/cm,因不同的材料而异),以诱导相邻原生质体局部接触区发生质膜的可逆击穿而形成融合。上述正弦波和方波脉冲均由自制的细胞融合仪发生。在本实验条件下,融合率可达50%以上,并探讨了方波脉冲讯号以及其它有关的条件对于融合率的影响。 相似文献
19.
20.
Efficient preparation of spheroplasts fromCandida utilis, Saccharomyces cerevisiae, andSchizosaccharomyces pombe, using a purified mixture of enzymes fromTrichoderma harzianum, is described. Limitations of other methods, and differences between yeasts are demonstrated. 相似文献