云南红豆杉愈伤组织诱导和组织培养

云南红豆杉愈伤组织的诱导频率为Ｗｈｉｔｅ〉ＭＳ〉Ｂ５,其中最高达８０％,叶片和种子之间差异不大,但不同时期的叶片差异较大,以嫩叶诱导为佳,激素配比以２,４－Ｄ２．０－３．０ｍｇ／Ｌ,ＢＡＰ０．５－１．０ｍｇ／Ｌ为宜,在继代培养阶段,Ｂ５比Ｗｈｉｔｅ和ＭＳ更适合细胞的快速生长、繁殖,并且有利于克服组织的褐化,适合于细胞生长的激素配比也以２,４－Ｄ２．０－３．０ｍｇ／Ｌ,ＢＡＰ０．５－１．０ｍｇ／Ｌｏ     

A Broad Spectrum Tissue Culture Experiment with Tobacco (Nicotiana tabacum) Pith Tissue Callus

The composition and rationale of a broad spectrum tissue culture experiment involving 81 different media are described. Tobacco calluses sub-cultured on media in this experiment were induced to form six main types of callus depending on the concentration levels of minerals, auxins, cytokinins, and sucrose, growth factors and amino acids in the medium. Only nine of the 81 media inhibited callus growth, and growth (increase in mass divided by initial mass of fresh matter) varied considerably reaching a maximum of nearly 200 after eight weeks on a medium high in minerals, cytokinins, sucrose, growth factors and amino acids and low in auxins. Five media induced regeneration from the calluses. This experiment is suggested as a potentially fruitful introductory test for many new or unresolved tissue culture situations.     

中药“枳壳”的组织培养与植株再生（简报）

以酸橙的茎段为外植体,进行离体再生体系研究.结果表明,培养基MS + 6-BA 1.0 mg/L + IAA 0.5 mg/L + CH 400 mg/L适宜不定芽诱导;培养基MS + 6-BA 0.5 mg/L+ IAA 0.25 mg/L + CH 400 mg/L适宜不定芽增殖;培养基1/2MS + NAA 1.0 mg/L + IBA 2.0 mg/L生根效果较好,生根率达80%以上.     

Morphogenetic Investigations in Callus Cultures of Tylophora indica

Callus cultures derived from stem explants of Tylophora indica (Fam. Asclepiadaceae) show a broad spectrum of tissue differentiation and regenerative potentialities over a wide range of nutrient conditions resulting in the production of roots, shoots, bipolar embryos and plantlets. This has been investigated and interpreted on a histological basis. Studies on the morphogenesis of embryos have shown that only such embryonal cells which were deeply embedded with in the callus parenchyma showed an ontogenic development reminiscent of ovular embryogeny whereas in those organized in the superficial layer of the callus the sequence of embryogeny was entirely different. Free cells growing in agitated liquid media first developed into filamentous structures and eventually, by repeated intercalary divisions, organized into bipolar embryos. Development of accessory embryos through cleavage and budding in the callus, differentiation of amphicribral vascular nodules, and meristematic colonies organized into spheres of parenchyma were also observed. These alternative morphogenetic events probably reflect the biochemical subtleties occurring within the callus as a result of an inadequacy or an imbalance of the various constituents in the culture medium.     

Cell Suspension and Callus Culture from Somatic Tissue of Maize

Cell suspension and callus cultures from somatic tissue of inbred lines of maize (Zea mays L.) were cultured on media that were defined via modification of a Linsmaier and Skoog preparation. Germlings incubated on the primary medium originally employed required long-term incubation for callus induction. Modification of the primary medium with high levels of iron and (ethylene dinitrillo)tetraacetic acid (EDTA), B vitamin amendments and vitamin E, shortened incubation by 75% and nearly doubled the percentage of germlings which produced callus. Callus did not remain viable in subcultures to the secondary medium originally employed, whereas a preparation, developed via modification of the original secondary medium, enabled perpetuation of callus through repeated subculture. Modification with high levels of iron and EDTA, plug B vitamins and vitamin E, with decreased concentrations of five inorganic salts, suppressed aberrant organogenesis and stabilized culture growth as viable callus. Similar modification, with the exception that EDTA was omitted, was employed for the development of a liquid medium. Tonicity of the medium was adjusted with a lowered level of sucrose, with the liquid further modified by addition of acetate. Upon development of this liquid, maize became the sixth monocot species for which somatic cells remain viable through repeated subculture in liquid suspensions.     

Tissue Culture of Maize I. Callus Induction and Growth

Callus induction and subculture was successful with mature embryos and stem sections of seedlings of Zea mays L. on Linsmaier and Skoog's medium modified to contain 4 mg/I of 2,4-D and 1 g/I of casamino acids. — 2,4-D was superior to NAA and IAA for both callus induction and growth. Callus subcultured on NAA formed abundant roots on agar-solidified media and numerous root-like primordia in liquid cultures. — Kinetin had no effect on callus induction in the presence of 2,4-D and neither kinetin nor gibberellic acid stimulated callus growth during subculture. — Callus grew equally well on the medium of Linsmaier and Skoog, that of Schenk and Hildebrandt, and the B-5 medium of Gamborg and Eveleigh containing 2% sucrose, 4 mg/I of 2,4-D and 1 g/I of casamino acids. — The callus grew more rapidly at 25°C than at 30°C or 35°C. Little difference was noted at any temperature in callus growth in alternating light (16 h) and dark (8 h) or continuous dark. — Sucrose was superior to glucose and maltose in both liquid and agar-solidified cultures. Lactose and galactose failed to support callus growth.     

伏令夏橙愈伤组织体细胞胚发生中多胺水平的变化

以继代培养8年的伏令夏橙愈伤组织为材料,研究了不同类型愈伤组织体细胞胚发生能力的差异和多胺水平的变化及两者之间的关系.结果表明,胚性愈伤组织的多胺含量高于非胚性愈伤组织,体细胞胚发生能力与多胺水平呈正相关.体细胞胚发生早期Put含量的增加有利于体细胞胚发生.球形胚大量形成时,Spd达到最高值;球形胚发育后期并有少量心形胚形成时Spm达到峰值.随着倍性的增加,伏令夏橙体细胞胚发生能力降低.精氨酸脱羧酶的活性变化与Put水平呈正相关,表明它是调节伏令夏橙体细胞胚发生中多胺水平的重要因子.     

药用植物佛甲草愈伤组织的诱导和培养

目的：佛甲草（Sedum lineare Thunb）是景天科植物中具有多种药用功能的植物,其肉质多浆的特点是愈伤组织成功诱导和培养的瓶颈。方法：以佛甲草的叶和茎为外植体,用正交试验对培养基中的激素浓度（2,4-D、6-BA和Kt）进行配置,在配置的18种培养基中筛选到适合佛甲草叶和茎愈伤组织诱导和继代的培养基配方。结果：MS＋3%蔗糖＋2,4-D2＋Kt1和MS＋3%蔗糖＋2,4-D2＋6-BA1培养基对茎和叶愈伤组织的诱导率分别达到70%和80%左右,而MS＋3%蔗糖＋2,4-D2＋6-BA1更有利于佛甲草愈伤组织的继代培养。结论：在愈伤组织培养中,佛甲草的茎和叶都可以作为优良的外植体进行愈伤组织的诱导和培养。实验结果为进一步通过组织培养手段提高佛甲草的药效成分提供了有益参考。     

Changes in Nuclear Ultrastructure during Callus Development in Tissue Culture of Allium Sativum

The changes in nuclear ultrastructure during garlic development were investigated. In quiescent cells, the nucleus was oblate and small, full of heterochromatin, and without nucleolus. In the active phase cells, the nucleus moved toward the center of the cell, and was enlarged with many deep fissures, and the heterochromatin in the nucleus turned to be euchromatin. One or more nucleoli were seen in active phase cells. Cell divisions were by means of amitosis. Relatively large nuclei with small distortions were seen in the callus cells. These results suggest that the metabolism of the active cells was higher than that of callus and quiescent cells. This revised version was published online in July 2006 with corrections to the Cover Date.     

Caffeine Formation in Tea Callus Tissue

Callus tissue derived from segments of the stems of the teaplant produced caffeine. The caffeine was found in the tissueand was also present in the growth medium. The amount of caffeinevaried with the duration and conditions of growth of the callus.Theobromine was also produced by the callus. The methylatedpurines appeared during the latter half of the period of therapid phase of growth. It is probable that the caffeine wasformed by those cells nearing the end of growth, maybe whenthe cells were autolysing. Suspensions of cells which were mechanicallybroken in a phosphate buffer at pH 7.2 produced more caffeineduring incubation at 26 ?C. If the autolysates were supplementedwith RNA and methionine the amount of caffeine produced couldbe increased. It is suggested that the caffeine is formed duringthe catabolic breakdown of nucleic acids rather than directlyfrom the pathways of purine synthesis.     

Citrus Tissue Culture : AUXINS IN RELATION TO ABSCISSION IN EXCISED PISTILS

An in vitro bioassay for chemicals that affect Citrus abscission was used to identify three inhibitors of stylar abscission in lemon pistil explants incubated on defined nutrient media. The three inhibitors (picloram, 4-chlorophenoxyacetic acid, and 3,5,6-trichloropyridine-2-oxyacetic acid) are all auxins, and the most potent of them (i.e. picloram) was found to be at least 10 times more active in the bioassay than 2,4-dichlorophenoxyacetic acid. Picloram (2 micromolar) also was shown to be effective in inhibiting stylar abscission in pistil explants from other Citrus cultivars such as mandarin, Valencia, and Washington navel oranges and grapefruit. To study the physiology of auxins active as abscission inhibitors versus inactive auxins in lemon pistils, the transport and metabolism of [1-¹⁴C]-2,4-dichlorophenoxyacetic acid was compared with that of [2-¹⁴C]indole-3-acetic acid, which is without effect in the bioassay over the range from 0.1-100 micromolar. Insignificant quantities of labeled indole-3-acetic acid and/or labeled derivatives were found to reach the presumptive zone of stylar abscission under the test conditions. Labeled 2,4-dichlorophenoxyacetic acid and/or labeled derivatives also were transported slowly through pistils, but some radioactivity could be detected in the stylar abscission zone as early as 24 hours after the start of incubation. Extensive conversion of [2-¹⁴C]indole-3-acetic acid to labeled compounds tentatively considered to be glycoside and cellulosic glucan derivatives was found with the use of solvent extraction methodology. A significantly smaller percentage of the radioactivity in pistils incubated on [1-¹⁴C]-2,4-dichlorophenoxyacetic acid was found in fractions corresponding to these derivatives. Both transport and metabolism appear to be important factors affecting the activity of auxins as abscission inhibitors in the bioassay.     

利用组织培养技术提取甘草黄酮

本研究筛选了两种能快速诱导甘草疯长型愈伤组织的培养基MS5和MS7,用微波法提取甘草愈伤组织和野生甘草根中总黄酮。测定结果显示:三年生野生型胀果甘草根中总黄酮的含量约为6.02%,培养3~4周的胀果甘草愈伤组织中总黄酮的含量约为1.1%。结果表明:组织培养生产黄酮在生产效率、工厂化生产、避免季节限制、保护野生甘草资源及实际应用中更具优势。本研究可为从甘草中提取总黄酮提供一条可借鉴的途径。     

Rearing Migratory Endoparasitic Nematodes in Citrus Callus and Roots Produced from Citrus Leaves

Radopholus similis and Pratylenchus coffeae were reared on callus and roots developed from citrus leaves. Callus formed best when leaf petioles were immersed in Astatula fine sand and the leaves were sprayed daily with 4 ppm 2,4-D solution and maintained at 25 or 30 C. The nematodes completed one generation in 20 days at 25 C. Highest populations of R. similis (1,127) occurred after 50 days, and the highest for P. coffeae (619) after 70 days. Leaf-callus cultures from R. similis-resistant citrus rootstocks showed the same degree of infection as susceptible rough lemon callus after 30 days.     

植物愈伤组织培养中内外源激素效应的研究现状与展望

综述了外源激素对愈伤组织诱导、分化的影响,以及内源激素在愈伤组织培养过程中的变化规律。有关愈伤组织培养中外源激素作用的研究日趋细化,而激素变化与适宜继代周期的关系已成为研究热点。随着分子生物学与同位素标记等技术的引入,必将促进愈伤组织培养中激素相关基因调控机理的揭示,以及激素作用位点的精确定位,从而推动植物愈伤组织培养的迅猛发展。