大豆组织培养再生植株

建立从离体培养高效率再生植株的程序是用细胞遗传操作改良农作物的一个先决步骤。从栽培大再组织培养能再生植株的报道,近年日渐增多(可参考文献)。其中,获得再生植株效率比较高的报道,所用的品种材料和     

THE BEHAVIOR OF TISSUE CULTURES FROM ENGLISH AND ALGERIAN IVY IN DIFFERENT GROWTH PHASES

In both the English and Algerian ivies, Hedera helix L. and Hedera canariensis Willd., leaf dimorphism of the juvenile and the mature, fruiting growth phases is pronounced, the former being a vine with lobed leaves and the latter a shrubby, upright form with entire leaves. Tissue cultures of English ivy started from stems of the two growth phases on the same plant consistently behaved differently, those from the juvenile stage invariably having the higher proliferation rate and larger cells. These differences were maintained consistently in monthly transfers over a period of two years. No medium was found which would support the growth of tissue subcultures of the adult stage of Algerian ivy, but all growth phases of the English ivy were cultured freely in a modified White's medium with additions of coconut water, naphthaleneacetic acid, enzymatic casein hydrolysate, and inositol. Cultures from individual open-pollinated seedlings of both species varied greatly in proliferation rate but were usually high.     

AXENIC TISSUE CULTURES FROM THE SPOROPHYTES OF LAMINARIA DIGITATA AND LAMINARIA HYPERBOREA (PHAEOPHYTA)

Starting material for the tissue cultures was the meristematic basal zone of the blade. Pieces treated 30–60 sec in hypochlorite solution were rinsed and placed on agar plates made from the artificial seawater ASP6 F2 solidified with 6 g agar l^?1. After 6 weeks colorless callus-like tissue grew out from some pieces. Treatment with activated charcoal removed some inhibiting substances from the agar medium as numbers of callus developing pieces increased on such plates. A combination of 10^?5 M NAA and 5 · 10^?7 M kinetin gave a yellow-brown tissue. A differentiation in the tissue from L. hyperborea was observed as well as the formation of meiospores, which grew out into male and female plants. Thalli of sporophytes were observed but they never reached a length of more than one mm before they died or changed to an irregular pattern of growth.     

ADVENTIVE ORGANOGENESIS FROM SOMATIC TISSUE CULTURES OF SOLANUM CAROLINENSE: ORIGIN AND DEVELOPMENT OF REGENERATED PLANTS

The development of adventitious shoot formation from cultured somatic tissues of Solanum carolinense was studied using light microscopy. For purposes of comparison, callus initiation and proliferation were also followed. When stem segments of this plant were cultured on medium supplemented with 6 mg/l benzyladenine (BA), cell division was first observed after 48 hr in the external phloem and inner cortex of the segments. This division activity gave rise to meristematic zones which subsequently formed shoot primordia within 6 days of culture. While organogenic potential appears to be limited to specific regions, all tissues of the explant were capable of callus formation when cultured on medium containing 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). In conjunction with a previous study, it appears that unlike somatic embryogenesis in this species, organogenesis does not require an intermediate callus phase for differentiation to occur.     

IONIC EFFECTS ON LIGNIFICATION AND PEROXIDASE IN TISSUE CULTURES

Crown-gall tumor tissue cultures release peroxidase into the medium in response to the concentration of specific ions in the medium. This release is not due to diffusion from cut surfaces or injured cells. Calcium, magnesium, and ammonium were, in that order, most effective in increasing peroxidase release. The enzyme was demonstrated cytochemically on the cell walls and in the cytoplasm. Cell wall fractions, exhaustively washed in buffer, still contained bound peroxidase. This bound peroxidase could be released by treating the wall fractions with certain divalent cations or ammonium. The order of effectiveness for removing the enzyme from the washed cell walls is: Ca⁺⁺ ≈ Sr⁺⁺ > Ba⁺⁺ > Mg⁺⁺ > NH₄⁺. These data support the thesis presented that specific ions can control the deposition of lignin on cell walls by affecting the peroxidase levels on these walls.     

红豆草组织培养物的超低温保存及其超微结构的观察

红豆草(Onobrychis viciaefolia Scop.)组织培养物在5%DMSO+10%甘油+8%蔗糖的冰冻保护剂及以1℃/分钟的速度降温到-35—-40℃,停留2小时后,投入液氮,40℃水浴快速化冻等条件下,存活率达60—70%,并保持了高的分化能力。电子显微镜的观察结果表明,快速冰冻和1℃/分钟慢速冰冻至-35℃—40℃不停留,对细胞结构造成严重的致死性破坏;-35℃停留30分钟对细胞结构的损伤是可逆性的;停留2小时的其超微结构基本上与对照材料无明显差别。     

CALCIUM AND THE CONTROL OF LIGNIFICATION IN TISSUE CULTURES

Lipetz , Jacques . (Rockefeller Inst., New York, N. Y.) Calcium and the control of lignification in tissue cultures. Amer. Jour. Bot. 49(5): 460–464. Illus. 1962.—Crown-gall tissues of Helianthus, Parthenocissus, Nicotiana, habituated Nicotiana, normal carrot, and Parthenocissus tissues grown in vitro on media low in calcium show an increase in lignification. The lignin is deposited not only on the walls of tracheids but on the walls of parenchymatous cells as well. Transferring the tissues to media containing higher calcium concentrations inhibits this excess lignification. Neither strontium nor magnesium could be substituted for calcium. These studies indicate that the availability and concentration of inorganic ions play a role in the control of lignification.     

植物组织培养物的超低温保存

引言十九世纪末,诞生了一门新的科学——低温生物学(Cryobiology)。气体液化技术使人类可以获得近乎生命的凝固状态。1949年发现了甘油可以防止冰冻对活细胞的伤害,对低温生物学有重要贡献。从而导致了在许多不同领域内广泛应用低温保护技术。超低温通常指低于-80℃的低温,常用的有干冰(-79℃)、深冷冰箱、液氮(-196℃)及液氮蒸汽相(-140℃)。这样低的温度下保存生物材料,可以大大减慢、甚至终止代谢和衰老过程,因而能     

THE USE OF TISSUE CULTURES TO PRODUCE VIRUS-FREE CLONES FROM INFECTED POTATO VARIETIES

By growing the excised apical meristems of sprouts from the potato varieties King Edward and Arran Victory, infected respectively with potato paracrinkle virus and potato virus S , virus-free plants were obtained. Although the method failed to produce virus-free plants from varieties infected with potato virus X , this virus also seems not to be present in apical meristems, for no virus could be demonstrated in callus tissue that developed from excised meristems less than 200 μ across. The concentration of tobacco mosaic virus in tomato roots and tobacco stems is also much less near the growing point than in older cells, but there is no evidence that the meristematic region is virus-free.     

SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM TISSUE CULTURES OF PENNISETUM AMERICANUM,AND P. AMERICANUM x P. PURPUREUM HYBRID

Immature embryos as well as explants obtained from young inflorescences of Pennisetum americanum (pearl millet) give rise to callus tissues on nutrient media containing 2,4-dichlorophenoxyacetic acid (2,4-D). A compact and pale-yellow callus that arises from the peripheral cells of the scutellum, and from the young inflorescences, undergoes further organized growth. When transferred to a 2,4-D-free medium, supplemented with indole-acetic acid or kinetin, or both, embryoids are formed in the organized areas of the callus. Embryoids show a bipolar organization with a shoot-coleorhiza (root) axis and have a coleoptile-like structure surrounded at the base by a cup-shaped structure that resembles the scutellum in texture and morphology. Embryoids show bilateral or radial symmetry and “germinate” in vitro to form plants that have been grown to maturity in soil. Similar embryogenic callus cultures have been produced from young inflorescence tissues of hybrid Pennisetum, a triploid sexually sterile hybrid of P. americanum x P. purpureum. Plants derived from these have also been transferred to soil. The regenerated plants showed normal chromosome numbers.     

甜菊组织培养物中叶绿体的超微结构与脱分代

含有叶绿体的甜菊（Ｓｔｅｖｉａｒｅｂａｕｄｉａｎａ）愈伤组织细胞转移至新鲜培养基后,导致光合片层的逐渐减少或消失,最后叶绿体脱分化形成原质体样的结构。超微结构观察表明,光合片层的减少或消失与降解及叶绿体分裂特别是不均等缢缩分裂而致基质组分和类囊体膜稀释有关。这一过程并不完全同步,一些质体含有少量正常的片展而另一些质体含有退化的片层甚至片展结构完全消失。细胞的一个明显特点是细胞器大多聚集在细胞核附近,细胞质增加并向细胞中央伸出细胞质丝。同时可观察到原质体。培养７ｄ后,许多细胞呈分生状态,细胞质富含细胞器,充满了细胞的大部分空间。此时细胞中的质体大多呈原质体状态。在细胞生长的稳定期,质体内膜组织成基质基粒片层,同时质体核糖体增加。文中讨论了高度液泡化细胞脱分化与细胞中叶绿体脱分化的关系。     

MORPHOGENETIC BEHAVIOR OF TOBACCO TISSUE CULTURES AND IMPLICATION OF PLANT SENESCENCE

Decreases in the growth and organ-forming capacities characterized continuously cultured tobacco (Nicotiana tabacum ‘Wisconsin 38‘) callus. The root-initiation ability was completely lost in 1 1/2-year-old cultures. The rate of shoot formation decreased to a low plateau in cultures that reached 1 1/2-3 years of age since the explanting. An inverse relationship between callus growth and in vitro clonal age was also observed. Studies with callus clones started from individually isolated pith cells showed that the growth and organ-forming potentials of somatic cells varied, signifying that cell alterations had occurred in vivo. Both totipotent and non-totipotent cell lines were obtained. Subculturing the single-cell lines through several passages disclosed that the morphogenetically depressed state was irreversible and instability was characteristic of the totipotent lines. In the latter, a change toward the morphogenetically repressed level was observed. These findings are discussed in relation to the phenomenon of senescence. It is suggested that an accumulation of somatic mutations, i.e., genetic alterations resulting in reductions in the morphogenetic potential of cells, is a basis underlying senescence.     

爬树龙的组织培养

爬树龙(Rhaphidophora decursiva (Roxb.)Schott),别名过山龙,系天南星科(Araceae)崖角藤属(Rhaphidophora)的一种藤本植物,以气根攀缘于它物上生长,叶表面绿色、发亮,它不仅是一种多年生常绿的观赏植物,而且其茎叶入药,可医治许多病症,因此有一定的药用价值。通常用扦插方法繁殖,我们取它的茎尖和茎节进行组织培养试验,现将初步试验结果简要报道如下。