不同培养条件对半夏悬浮培养细胞生长及总生物碱形成的影响

研究了半夏悬浮培养过程中细胞鲜重的变化和MS培养基组分中碳源、钙盐、Fe盐及肌醇变化对半夏悬浮培养细胞生长及总生物碱合成的影响。结果表明 ,半夏细胞的生长曲线呈“S”型 ,细胞的最佳收获时间为 2 1d。在两种碳源中 ,葡萄糖比蔗糖利于细胞的生长和总生物碱的合成 ,最适浓度为 2 0g L。最适合细胞生长和总生物碱形成的钙盐浓度为 1 8mmol L、Fe盐浓度为 0 0 6mmol L和肌醇浓度为 10 0mg L。     

Effects of Culture Conditions on Tropane Alkaloid Formation in Hyoscyamus niger Suspension Cultures

Tropane alkaloid formation was studied under various culture conditions in suspension cultures of Hyoscyamus niger L. High aeration increased cell growth and the contents of hyoscyamine and scopolamine. White’s medium inhibited cell growth but increased the hyoscyamine content. The low concentrations of nitrogen and phosphate in White’s medium were responsible for such effects. NAA promoted cell growth but inhibited hyoscyamine formation. A hyoscyamine content of 0.05% dry weight (DW) (0.14 mg in 25 ml of culture medium) was achieved in modified LS medium in which the phosphate concentration was decreased to 0.2 mm and the auxin omitted.

Effects of precursor feeding on alkaloid formation were also studied with the putative precursors of hyoscyamine. N-Methylputrescine, tropine, phenylalanine, and tropic acid increased alkaloid formation, but the promotive effect of these precursors varied from one experiment to another.     

Effects of Chemical and Physical Conditions on Growth of Camptotheca acuminata Cell Cultures

Callus was induced from Camptotheca acuminata, which produces an antitumor alkaloid, carnptothecin. Using the Murashige and Skoogs’ medium as the basal, cultural conditions were examined for C. acuminata suspension cultures. As a result, a medium, containing 0.1 mg/liter 2,4-D, 3 mg/liter kinetin and 0.05 mg/liter GA₃, was established as a medium that gave the best cell growth in suspension cultures. In addition, conditioning of medium and addition of 0.115 mm l-Trp and l-Phe to medium promoted remarkably growth of cell suspensions.     

不同培养条件对光叶楮试管苗玻璃化的影响

以MS为基本培养基,研究生长调节剂、蔗糖、琼脂、光照强度和温度等因素对光叶楮试管苗玻璃化的影响。结果表明,最佳的生长调节剂配比是1.0 mg/L 6-BA、0.3 mg/L NAA,蔗糖浓度3.5%,琼脂浓度0.35%,光照强度2 000 lx,培养温度25℃。该优化培养条件能有效防止玻璃化苗的发生,获得较高的增殖系数和较健壮的苗木,且能降低生产成本。     

Initiation and Growth of Tissue Cultures of Theobroma cacao

Difficulties were experienced in obtaining sterile explant materialfrom which to initiate the callus cultures. The material whichprovided the most effective source in terms of numbers of explants,levels of contamination and ease of sterilization was from seedlingtissue, grown from seeds which had been transported in the intactpod. The seeds were removed asceptically and germinated in sterileflasks on White's medium solidified with agar, then at 3–4weeks old the seedling was sectioned into root, stem, cotyledon,hypocotyl and apical bud explants. The medium which promotedthe most rapid and sustained callus growth was a modified Murashigeand Skoog medium. Callus transferred to a liquid medium formeda cell suspension which by selecting for a low level of aggregationand a fast growth rate eventually developed these characteristics.There was evidence for clonal differences in the callus basedon growth rate data and appearance and related to the originof the initial explant tissue. Although the callus was exposedto considerable variation in the nutrient medium, except forthe occasional appearance of root-like structures, no regularorganogenesis was found.     

Investigations on Growth and Metabolism of Plant Cells: VII. Sources of Nitrogen for Tissue Cultures under Optimal Conditions for Their Growth

Tissue cultures from explants of carrot root and potato tuber,stimulated into rapid growth by the addition of coconut milkorAesculus liquid endosperm to the medium, become to a certaindegree heterotrophic for nitrogen. Maximum growth-rates areattained only when nitrate is supplemented with various reducednitrogen compounds. The effects of casein hydrolysate, amino-acidmixtures, ammonia, tryptophan, urea, and allantoin have beeninvestigated, and their possible biochemical roles are discussed.     

Growth and Morphogenesis in Tissue Cultures of Anagallis arvensis

Morphogenesis has been induced in excised organs and callus tissue cultures obtained from various parts of the seedling and mature plants of pimpernel (Anagallis arvensis). Vigorously growing cell cultures capable of being periodically subcultured have been established in liquid as well as on the agar-solidified Murashige and Skoog's medium supplemented with 2,4-D (0.1 mg/1) + kinetin (0.1 mg/l) + coconut milk (10%). The callus tissue obtained from excised hypocotyl segments is white, soft, friable and fast growing, and has been subcultured over a period of two years without showing any sign of decline in growth. The optimum conditions for growth are at pH 5.9, temperature 27°C, and with 4% sucrose as the carbon source. Under appropriate nutritional supply these cultures can be manipulated to induce rhizogenesis in the suspension cultures, and buds and “embryo-like” structures on agar-solidified media. The excised leaves, hypocotyl and stem segments regenerate buds. Of the cytokinins used, 6-(y,y-dimethylallylamino)-purine proved to be the best for the number of cultures producing buds, as well as for the number of buds per culture. Anatomical studies revealed that buds arise from the epidermal and subepidermal layers of leaves and hypocotyl; these buds form shoots which eventually develop into plantlets.     

Improved Growth of Tissue Cultures of the Onion, Allium cepa

A forty-fold increase in tissue fresh weight has been achieved for Allium cepa stem tissue callus grown on a modified B5 medium with increased ammonium, phosphate and nitrogen levels; and decreased level of 2,4-dichlorophenoxyacetic acid. Growth was examined on several standard media, using fresh weight, dry weight and cell number as parameters over an 8-week growth period. Further, tissue grown upon this medium is more friable than on other media. The value of such friable callus for suspension culture initiation and for single cell cloning experiments is discussed.     

Growth and Cytology of Celery Plants Derived from Tissue Cultures

Callus was initiated from petiole explants of 3-month-old celeryplants and sub-cultured at monthly intervals on a modified Murashigeand Skoog medium. 6 months later a further batch of callus wasinitiated, again from 3-month-old plants. When the first batchof callus had been sub-cultured for 15 months, and the secondbatch for 9 months, the callus from both sources was placedon a hormone-free medium to allow plants to form. Normal seedlingswere grown at the same time and when the plantlets were largeenough both the plantlets and the seedlings were transferredto individual pots and grown to maturity in a greenhouse. Thepetiole height, width and number of petioles were measured atintervals during the growing period and at one stage root tipswere excised from a number of plants in each group and the chromosomenumber counted. The growth and cytological data showed thatthe plants derived from tissue culture were not affected bydifference in sub-culture time and were very similar to theseedling plants.     

表油菜素内酯对苦参愈伤组织生长的影响

在苦参愈伤组织生长的最佳培养基:改良MS+TDZ 1.0 mg/L+2,4-D 0.5 mg/L+Cys20 mg/L上分别添加表油菜素内酯(epibrassinolide,epi-BR)0、10-7、10-5,10-3、10-1 mg/L.结果表明,愈伤组织鲜重随epi-BR浓度升高而升高,呈正相关,而干重则在10-3mg/L epi-BR浓度下达到最高,之后随epi-BR浓度升高呈下降趋势.     

Growth of Mixed Cultures on Mixed Substrates: I. Continuous Culture

Continuous culture on mixed glucose-lactose or glucose-butyrate media inoculated with river water led to a population composed of a pseudomonad and a coliform. The glucose was used preferentially to the other carbon source, and the utilization of the secondary carbon source was greatly reduced at high growth rates. Significant amounts of acetate were excreted even though the cultures were limited by the carbon source, rather than by oxygen or other nutrients. At high growth rates, the pseudomonad dominated the population, whereas at low and moderate growth rates the coliform was dominant. A syntrophic relationship was shown by the fact that the pseudomonad could not grow alone on the glucose-butyrate medium.     

地生弯颈霉的培养与生长条件

从长白山北坡森林土壤中分离纯化得到1株地生弯颈霉(Tolypocladium geodes),对其培养及生长条件(如培养基、光照、温度、碳源、氮源和pH等)进行了试验。结果表明,地生弯颈霉在麦芽浸膏琼脂培养基(MEA)上菌落生长直径最大,马铃薯葡萄糖琼脂培养基(PDA)上产孢量最高;5~30℃温度范围内均能生长,25℃时生长和产孢量最佳;最适碳源和氮源分别为葡萄糖和柠檬酸铵;光暗交替条件下,最利于其生长和产生孢子;pH4~9范围内均能生长和产孢,pH=8时菌落直径最大,pH=6时产孢量最高。     

Effect of 5-Fluorouracil on Growth and Morphogenesis of Tissue Cultures of Nicotiana sylvestris

Tissue cultures of Nicotiana sylvestris treated with increasingconcentration of 5-fluorouracil (5-FU) showed a differentialeffect of the drug on growth and morphogenesis. In the rangeof concentrations tested, 5-FU did not inhibit increases infresh weight, but it inhibited the production of shoots. Theeffect of 5-FU on shoot production was reversible and dependedon the time of adding 5-FU to the tissue culture. Uracil andthymine did not counteract the effect of 5-FU on morphogenesis,whereas uracil partially reduced the inhibition of growth undersome culture conditions. (Received July 16, 1985; Accepted April 5, 1986)     

Effect of Culture Origin and Conditions on Duvatrienediol Accumulation in Shoot Cultures of Tobacco

A comparison of morphology, trichome type and duvane accumulationof tobacco shoot cultures originating from seedlings, callusand cell suspension cultures incubated on a range of cytokininconcentrations is presented. A method for the extraction andHPLC analysis of duvatrienediols from cultures is described.Duvatrienediols (DVTs) were not detected in callus or cell suspensioncultures and were only detected in seedling-denved shoot cultures.Shoot development was dependent on explant type probably asa result of pre-exposure of callus and suspension cells to highauxin concentrations. Low cytokinin levels promoted shoot developmentwhilst high cytokinin levels increased the number of shootsregenerated but retarded their development. DVT accumulationoccurred only in ‘normally’ developed shoots withtrichomes. Growth of shoot cultures in liquid medium was inhibitoryto the accumulation of DVTs. Optimum conditions for the accumulationof DVTs were shown to be incubation of seedling-derived shootcultures on media containing 10^–6 M BAP solidified withagar. The yield was 120 µg g^–1 fr. wt. which iscomparable to greenhouse-grown material. Both and ß-DVTwere detected in a ratio of 5: 1. The relationship between tnchomesand DVT accumulation is discussed with reference to the rolesof culture origin, cytokinin, trichome type and environmentaleffects. Key words: Tobacco, duvatrienediol, trichome, shoot culture