凹叶厚朴组织培养的研究

采用单因子比较、正交设计、均匀设计法，分别研究了影响凹叶厚朴Ｍａｇｎｏｌｉａｂｉｌｏｂａ愈伤组织诱导和生长的各种因素。研究结果表明：愈伤组织诱导的最适培养基为Ｂ５＋２，４－Ｄ２ｍｇ／Ｌ＋６－ＢＡ１ｍｇ／Ｌ，生长的最适培养基为Ｂ５＋２．４－Ｄ１ｍｇ／Ｌ＋６－ＢＡ１．５ｍｇ／Ｌ。在培养基中添加１０％豆芽汁能够促进愈伤组织迅速生长。黑暗培养对愈伤组织的生长极为有利     

群众杨39无性系耐盐悬浮细胞系的建立和体细胞变异体完整植株的诱导*

本试验以群众杨３９号为试材，通过耐盐胁迫悬浮培养建立耐盐悬浮细胞系，经愈伤组织成功培育出耐ＮａＣｌ３．０‰～３．５‰的群众杨体细胞变异体的完整植株。实验表明，ＭＳ培养基附加０．４５ｍｇ／Ｌ２，４－Ｄ、０．３ｍｇ／ＬＮＡＡ和０１ｍｇ／ＬＫｉｎｅｔｉｎｅ的Ｍ４培养基能较好地获得松脆、易分散的愈伤组织。液体以ＭＳ培养基只附加０．５ｍｇ／Ｌ２，４－Ｄ的ＬＭ３为最好，附加氨基酸有益于悬浮细胞的正常生长。还对悬浮培养细胞的有关参数进行了测定。耐盐悬浮细胞培养实验结果表明ＮａＣｌ对细胞生长有抑制作用，并随着ＮａＣｌ升高而加强。对获得的耐３‰～６‰各水平ＮａＣｌ悬浮细胞经高密度植板，都能形成愈伤组织。耐盐愈伤组织诱导只获得耐３‰～４‰ＮａＣｌ的不定芽，高于４‰ＮａＣｌ未能诱导出不定芽，说明高浓度ＮａＣｌ对芽组织分化有明显的抑制作用。不定根分化对ＮａＣｌ反应非常敏感，只在耐３‰和３．５‰ＮａＣｌ浓度培养基诱导出不定根。     

凤尾丝兰子房离体培养中的激素效应研究

用ＭＳ培养配方为基本培养基，附加ＫＴ、６－ＢＡ、２。４－Ｄ、ＮＡＡ及ＩＢＡ等５种植物激素按不同水平与组合方式，以凤尾丝兰离体子房为试验对象，研究其在培养过程中各分比阶段的激素效应。结果表明：脱分化，再分化及生根诱导各阶段，细胞激动素与生长素的种类、性质、绝对浓度及配合比例至关重要。ＫＴ３～５ｍｇ／Ｌ＋２．４－Ｄ０．１～０．２ｍｇ／Ｌ可诱导愈伤组织产生，但以ＫＴ４ｍｇ／Ｌ＋２．４－Ｄ０．１ｍｇ／Ｌ效     

几种荫生观叶植物芽器官诱导培养

用比鲁逊白掌（Ｓｐａｔｈｉｐｈｙｌｌｕｍ“Ｉｌｌｕｓｉｏｎ”）超级白掌（Ｓ．“Ｓｕｐｐｅｒ”）、丹尼尔白掌（Ｓ．“Ｄａｎｉｅｌ”）、特纳万年青（Ｄｉｅｆｆｅｎｂａｃｈｉａ“Ｔｅｒｎａ”）芽器官为外植体，通过组织培养诱导出不定芽。比鲁逊白掌、超级白掌、丹尼尔白掌始芽诱导出正常芽丛较好的培养基为：Ｍｓ＋ＢＡ２～３ｍｇ／Ｌ＋ＮＡＡ０．５ｍｇ／Ｌ；特纳万年青始芽诱导出正常芽丛较佳的培养基为：Ｍｓ＋ＢＡ２～３ｍｇ／Ｌ＋ＩＡＡ０．５ｍｇ／Ｌ，其第１～２个腋芽的繁殖系数较高２．０。     

苦丁茶的组织培养研究

诱导苦丁茶具节茎段芽体发生、生长与增殖的培养基,以改良ＡｎｄｅｒｓｏｎＭＳ培养基＋ＢＡ２．０ｍｇ／Ｌ＋ＮＡＡ０．１ｍｇ／Ｌ＋ＺＴ２．０ｍｇ／Ｌ＋ＧＡ３０．５ｍｇ／Ｌ较为适宜;以改良ＡｎｄｅｒｓｏｎＭＳ培养基＋ＢＡ３．０ｍｇ／Ｌ＋ＮＡＡ０．２５ｍｇ／Ｌ＋ＺＴ１．０ｍｇ／Ｌ继代培养。培养时温度为２８℃,光照度为１５００ｌｘ,光照时间为１４ｈ／ｄ;生根处理用１／４ＭＳ＋ＩＢＡ１．０ｍｇ／Ｌ培养基。     

蓝桉组织培养的研究

用蓝桉（Ｅｕｃａｌｙｐｔｕｓｇｌｏｂｕｌｕｓ）离体芽器官诱导培养，分化形成丛生芽，年繁殖系数３￣（１２）。０．１～０．５ｍｇ／Ｌ的６－ＢＡ或０．５～０．８ｍｇ／Ｌ的ＫＴ诱导外植体（带节茎段）腋芽萌动的效果最佳，诱导率分别达８０．３％和８１．５％。１．５～２０ｍｇ／Ｌ的６～ＢＡ或２０～２．５ｍｇ／Ｌ的ＫＴ分别与０．５～１．０ｍｇ／Ｌ的ＮＡＡ组合，对于促进腋芽分化形成丛生芽及继代培养中芽的增殖具有最佳效果。培养基中的无机盐浓度、蔗糖含量对蓝桉试管苗的生根具有显著影响；ＩＢＡ促进蓝桉试管苗的生根。至目前为止，在１／２ＭＳ无机盐培养基＋ＩＢＡ１．２～１．４ｍｇ／Ｌ＋Ｓ５ｇ／Ｌ中诱导生根，生根率最高可达２６．４％。     

赤桉下胚轴不同再生途径建立研究

赤桉下胚轴不同再生途径建立研究表明，Ｂ５为基本培养基添加２，４－Ｄ１－２ｍｇ／Ｌ及２，４－Ｄ１－２ｍｇ／Ｌ附加ＫＴ０．５ｍｇ／Ｌ可以诱导出愈伤组织，诱导率达１００％；６ＢＡ０．５ｍｇ／ｌ与ＮＡＡ０．２ｍｇ／Ｌ配合使用，可使胚状体发生率达４２．８％。ＮＡＡ３ｍｇ／Ｌｇｎ ６ＢＡ０．２ｍｇ／Ｌ的结合愈伤诱导率也为１００％，例题胚状体发生率只有１７．５％，器官发生率７５．０％。ＡｇＮＯ３０．０５％可促进     

群众杨悬浮细胞系的建立和耐盐体细胞变异体的初步筛选

对群众杨首次进行了悬浮细胞培养，对悬浮细胞系的生长进行测定，同时对耐盐体细胞变异体进行了初步筛选，获得了耐３．０‰－４．０‰不同ＮａＣＬ水平的不定芽。获得适宜悬浮、松脆、易分散的愈伤组织是悬浮的关键，对不同供试脱分化激素配比的试验结果表明，诱导愈伤组织的ＭＳ＋２，４－Ｄ（０．４５ｍｇ／Ｌ）＋ＮＡＡ（０．３ｍｇ／Ｌ）＋Ｋｉｎｅｔｉｎ（０．１ｍｇ／Ｌ）为最好。合适的液体培养基成份和培养技术也是悬浮细胞     

山影拳的组织培养

文章着重介绍了山影拳组织培养中的愈伤组织的诱导及芽分化的过程。实验结果表明，在ＭＳ＋６－ＢＡ０．５ｍｇ／Ｌ＋２，４－Ｄ２ｍｇ／Ｌ＋ＮＡＡ０．１ｍｇ／Ｌ＋３０ｇ／Ｌ蔗糖＋１０ｇ／Ｌ琼脂的培养基中，山影拳的茎段切面上可以很快诱导出愈伤组织，愈伤组织白色、疏松、颗粒状；在ＭＳ＋６－ＢＡ０．５ｍｇ／Ｌ＋ＮＡＡ０．１ｍｇ／Ｌ的培养基上可诱导出呈浅绿色的愈伤组织，经一段时间以后，可分化出不定芽或在外植体的突起     

柚木优树的快速繁殖

在越南胡志明市进行柚木选优和优树快速繁殖，其个植体选用优树萌芽条件扦插萌生的腋芽茎 和茎尖。结果表明，芽增殖培养基为ＭＳ附加６－ＢＡ，２．０ｍｇ／Ｌ，ＫＴ１．０ｍｇ／Ｌ和ＮＡＡ０．５ｍｇ／Ｌ，生根培养基以１／２ＭＳ附加ＩＢＡ１．０ｍｇ／Ｌ，ＬＡＡ０．５ｍｇ／Ｌ效果最好。     

Somatic embryogenesis from cell suspension cultures of aspen clone

Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions.     

Establishment of cell suspension line of <Emphasis Type="Italic">Populus tomentosa</Emphasis> Carr.

Leaves of fine Populus tomentosa genotype TC152 were used as explants to establish cell suspension lines. The effects of plant growth regulators on callus induction and establishment of cell suspension lines were studied. The callus induction rate was the highest on a MS solid medium supplemented with 1.0 mg·L^-1 2,4-D. A cell suspension line could be obtained by inoculating calli which were not subcultured into a MS liquid medium supplemented with 1.5 mg·L^-1 2,4-D. The best subculture medium was MS ＋ 0.8 mg＇L-1 2,4-D ＋ 30 g·L^-1 sucrose with a subculture cycle of seven days.     

Callus initiation and subculture ofTaxus chinensis

Conditions have been established for the callus initiation and subculture ofT. chinensis. The calli were induced by the explants cultured first on the medium MS supplemented with 1.0 mg/L 2,4-D, 2g/L CH, and 25g/L sucrose, then on The medium: MS+1.0 mg/L NAA+0.5 mg/L BA+2g/L CH+25g/L sucrose. When the callus was subcultured and tamed several times, it could grow fast and stable on the medium: MS+0.2mg/L 2,4-D+0.5 mg/L NAA+0.5 mg/L BA+2g/LCH+25 g/L sucrose. The contamination of explants was a result of endophytic microbes ofT. chinensis. This could be avoided by adopting the tender shoots 3–5 cm long collected in early spring as the source of explants. The browning of the cultures could be prevented and controlled by means of the selection of a suitable explants, hormonal regime in the medium, culture methods and the use of antioxidants. Responsible Editor: Chai Ruihai     

Somatic embryogenesis and plant regeneration inAcanthopanax sciadophylloides

Somatic embryos ofAcanthopanax sciadophylloides Franch. et Sav. were differentiated from both zygotic and somatic embryos and calli, and plants were regenerated from these somatic embryos. A zygotic embryo, enclosed within a small portion of the endosperm, was incubated on Murashige and Skoog (MS) media supplemented with various combinations (range 0–10.0 mg/l) of 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D). After 4 months, swelling of the zygotic embryos and callus formation was observed. When the swollen embryos were transferred to MS medium supplemented with 0.5 mg/l of 2,4-D, somatic embryos were formed in one to two months. After subculture on the same medium, new embryos were differentiated from various parts of the older somatic embryos. The calli were cultured on medium supplemented with 2.0 mg/l of 2,4-D and BAP for three weeks. Proliferated calli were transferred to medium supplemented with 1.0 mg/l of 2,4-D and BAP. Somatic embryos were differentiated from the calli within one to two months. Somatic embryos were germinated on half-strength MS medium without plant growth regulators and the plantlets were grown in soil. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995) & First Asia-Pacific Symposium on Forest Tree Genetic Improvement (Beijing).     

Plantlet regeneration from leaf protoplasts ofPopulus euphratica

Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the cell density of 9×10⁴/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of 2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and plantlets were regenerated on 1/2 MS medium without a hormone. A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995).     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Caulogenic callus induction and adventitious bud formation from embryos of long-term stored seeds ofPicea jezoensis

Caulogenic calli with a high differentiation potency were induced from mature embryos ofPicea jezoensis seeds stored over a long time, for 29 years, resulting in the active formation of adventitious buds. Embryos began to induce calli within 3 weeks of cultivating on LP medium containing 3 μM BAP and 1 μM 2,4-D. Then, the calli proliferated and transformed into caulogenic calli with bud primordia in 8 weeks. The caulogenic calli increased actively with the addition of 500 mg/l ofl-glutamine in the medium. Furthermore, caulogenic calli, induced on LP medium containingl-glutamine, resulted in the formation of adventitious buds, which elongated after transferring the calli into LP medium with 0.1 μM BAP, but withoutl-glutamine. It appears that the number of adventitious buds and the process of shoot elongation are influenced by the kind of nitrogen contained in the medium for callus induction. A part of this study was presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

<Emphasis Type="Italic">Nitraria sibirica</Emphasis> cell suspension culture: establishment,characterization and application

Nitraria sibirica Pall. is a shrub that grows in saline-alkali soil and has traditional medicinal value and potential commercial value. The objectives of this study include induction and multiplication of callus, establishment of a suspension cell line, and isolation of protoplasts from cell suspensions. Murashige and Skoog (MS) medium was used for callus induction from mature seeds of N. sibirica. Seed-derived calluses were further multiplied on MS medium augmented with 0.5 mg L^?1 6-benzylaminopurine (6-BA) and 1.0 mg L^?1 2,4-dichlorophenoxy (2,4-D) acetic acid. Suspension cultures of N. sibirica were initiated by transferring friable calli to the same liquid multiplication medium. Characterization of the suspension culture was assessed based on fresh mass, dry mass, cell viability and pH value of the culture. A typical growth curve was observed after inoculating 1.5 g of callus in 40 mL liquid medium, including a lag phase, an exponential growth phase, a stationary phase, and a negative acceleration phase. The effect of factors such as pre-plasmolysis, enzyme combination, enzymolysis time and mannitol concentration, on the isolation of cell-derived protoplasts were evaluated to determine the usefulness of suspension cultures. The maximum yield (9.79 × 10⁶ cells/g) and highest viability (79.97%) of protoplast were reached when approximately 1 g of cell suspension (cultured for 6 days) was inoculated for 12 h in cell and protoplast washing solution made of 0.8 mol L^?1 mannitol mixture solution, cellulose onozuka R-10 2% (w/v), hemicellulose 0.2%, macerozyme R-10 1%, and pectolyase Y-23 0.5%. Protoplast yield was significantly influenced by pre-plasmolysis and cellulose onozuka R-10 (P < 0.05).