植物原生质体培养方法

1　植物原生质体培养的简史植物细胞原生质体 ,在植物学上指植物细胞通过质壁分离后 ,可以和细胞壁分开的那部分细胞物质。原生质体分离纯化后 ,须在适当的培养基上应用适当的培养方法 ,才能再生细胞壁 ,并启动细胞持续分裂 ,直至形成细胞团、长成愈伤组织或胚状体、分化和发育成苗 ,最终再生完整植株。其中 ,选择合适的培养方法始终是原生质体培养中最基础也是最关键的一环。植物原生质体培养方法起源于植物单细胞培养方法。早在 190 2年 ,Haberlant通过实验就预言 :体外培养单个细胞可通过其分裂得到培养组织。直到 1954年 ,植物单细胞培…     

菊科植物原生质体研究进展

介绍了目前菊科植物原生质体研究进展,重点对菊科植物原生质体分离、培养、影响原生质体再生的因素、原生质体再生植株的变异、原生质体的应用等方面的研究工作进行了总结,提出了存在的问题和今后的工作重点。     

植物原生质体培养及有关生理基础问题

植物原生质体培养及有关生理基础问题赵颖,梁海曼（杭州大学生物系,杭州３１００１２）ＰＬＡＮＴＰＲＯＴＯＰＬＡＳＴＣＵＬＴＵＲＥＡＮＤＳＯＭＥＢＡＳＩＣＰＨＹＳＩＯＬＯＧＩＣＡＬＰＲＯＢＬＥＭＳ￥ＺｈａｏＹｉｎｇａｎｄＬｉａｎｇＨａｉｍａｎ（Ｄｅｐａｒ...     

植物防御反应与原生质体培养技术的改进

介绍了脱壁酶分离原生质体操作过程中,植物细胞防御反应的发生及其对原生质体分离与早期阶段培养的影响;阐述了原生质作早期阶段培养的实质是遭受机械、生理损伤的植物细胞完成其自身修复的过程;并对改进原生质体培养技术进行了分析。     

核果类果树原生质体培养研究进展

本文主要介绍了核果类果树原生质体培养的材料来源、分离培养、植株再生,简要介绍了其杂交和遗传转化,并对以后的工作进行讨论。     

植物原生质体培养研究进展及应用

综述了原生质体培养的研究进展。原生质体培养在理论研究上的应用有３个方面：（１）细胞生理现象的研究;（２）细胞与细胞质相互关系的研究;（３）病毒侵染与复制机理的研究。在原生质体培养中可通过遗传操作和突变体的筛选对品种进行改良,创造优异品种。     

植物生物技术讲座（一）：原生质体培养

植物原生质体是指通过质壁分离,能够和细胞壁分开的那部分细胞物质。换言之原生质体就是除去全部细胞壁的“细胞”,或是一个为原生质膜所包围的“裸露细胞”。植物原生质体由于失去细胞壁,更由于单个原生质体即包含一个物种的     

植物原生质体培养研究新进展

近几年来植物原生质体培养取得较大进展;重要农作物如水稻、小麦、棉花、玉米、栽培大豆等都有了突破,试验技术和方法也有不少改进。本文介绍有关的主要结果并讨论存在问题和发展趋势。     

禾本科植物原生质体培养及研究进展

植物原生质体培养是高等植物细胞工程和基因工程的重要基础之一。许多遗传操作,如体细胞杂交、细胞质重组和直接的 DNA 摄入等,都依赖于原生质体的再生。这种再生在双子叶植物中已不是一件困难的事。然而,多年来禾本科植物原生质体培养一直被认为是个十     

Progress in plant protoplast research

During the past years plant regeneration from protoplasts was achieved for a number of important crops (maize, sorghum, rice, wheat, sugar beet). The use of embryogenic tissue for protoplast isolation greatly contributed to this success. There was also some progress in woody plant species and ornamentals. Fusion of protoplasts resulted in may fertile hybrid plants, especially in the Brassicaceae and Solanaceae. These somatic hybridization studies led to introduction of new agronomical traits from sexually incompatible species into the cultivar gene pool and to new nucleus-organelle compositions. The limitations of somatic hybridization, mainly imposed by the taxonomic distance of the parents, and expressed as chromosome loss and reduced fertility, are more clearly recognized now. Asymmetric hybridization with irradiated donor protoplasts resulted in cybrids with new cytoplasmic traits (e. g. intraspecific fusions in Brassica ), as well as in the transfer of only a few donor choromosomes (e. g. intrageneric fusions in Nicotiana ). Most intrageneric fusions, however, resulted only in a limited elimination of donor chromosomes (e. g. in Lycopersicon ), and polyploidization occurred (e. g. in Nicotiana ). Also some success on protoplast transformation was obtained in both monocots and dicots. Fertile transgenic rice plants (Japonica, Indica) were produced after direct gene transfer into protoplasts derived from embryogenic cell suspensions. Particle gun experiments using embryogenic cell suspension of maize resulted in fertile transgenic plants. Transformation of citrus and lettuce by direct gene transfer was also reported.     

Cauliflower inflorescence protoplast culture and plant regeneration

A yield of 2–4×10⁶ protoplasts/g F.W. could be obtained when fresh cauliflower inflorescence segments were digested with 2% cellulase Onozuka R-10, 1% cellulase RS and 0.4% Macerozyme R-10 in CPW18S for 7 to 10 h. Purified protoplasts were cultured in K8p liquid and agarose medium. Although protoplasts in liquid medium divided earlier than in agarose, protoplast-derived cells in liquid culture could not avoid browning. With agarose culture, sustained division and callus formation could be achieved. After 20 days, calli were transferred onto B5 agar medium with ZT 1.5 mg l^-1, BA 0.5 mg l^-1 and IAA 0.1 mg l^-1 for shoot formation. The frequency of bud formation varied from 56.7% for calli of 1mm in size to 5.6% for 5mm calli. The shoots formed were rooted in B5 medium containing 0.5 mg l^-1 IBA, and the regenerated plants were transplanted to pots and grew normally. It took about two months from protoplasts to the regenerated plants.Abbreviations Ade adenine - BA 6-benzyl aminopurine - CH casein hydrolysate - CM coconut milk - 2,4-D 2,4,-dichlorophenoxyacetic acid - GA₃ gibberellic acid - Gln glutamine - NAA -naphthylacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - ZT zeatin     

An improvement of tomato protoplast culture for rapid plant regeneration

Tomato mesophyll protoplasts were cultured in TM2 medium containing 5.7 M -naphthaleneacetic acid and 2.4 M benzyladenine and were incubated either in stationary culture or on an orbital shaker at 25–30 strokes per min, in combination with interval addition of fresh medium. The effects of stationary and shaking conditions on the growth of the colonies and their subsequent shoot organogenesis were significantly different. The cultures maintained in stationary condition without adding fresh medium accumulated a thin membranous layer on the medium surface and whitish substance in the medium that seemed to precede cell browning and premature colony death. Mild shaking conditions along with the reduction of colony density by one half by dividing the contents of one dish into two dishes, after adding 2 ml of fresh medium on the 4th day and further addition of fresh medium (0.5 ml) on the 8th day of plating, provided optimal conditions for colony growth and suppressed thin layer and whitish substance accumulation. Ten-day-old colonies raised through this protocol regenerated shoots rapidly (within 19–20 days after initial plating) after transfer to regeneration medium (MS medium with 2.8 M zeatin riboside, 0.06–0.1 M gibberellic acid, 4% sucrose and 1% type VII agarose) directly bypassing the callus phase.Abbreviations BA benzyladenine - GA₃ gibberellic acid - IAA indoleacetic acid - MS Murashige & Skoog (1962) medium - NAA -naphthaleneacetic acid - SPM stroke per min - GLM General Linear Models - 2,4-D 2,4-dichlorophenoxyacetic acid     

Studies of protoplast culture and plant regeneration from commercial and rapid-cyclingBrassica species

Protoplasts were isolated from aseptic shoot cultures of commercial cultivars ofBrassica napus, B. oleracea andB. campestris, and from the six rapid-cycling brassica species. Of the rapid-cycling species, onlyB. napus responded well to the culture conditions used; 2% of protoplasts formed calli and up to 5% of calli regenerated shoots. Regeneration was also achieved from commercial cultivars ofB. napus andB. oleracea. For these two species the plating density, time of dilution with fresh medium and the composition of the shoot-inducing medium were all found to have an important influence on the efficiency of plant regeneration. Both responded better to maltose than to sucrose-based media. Under the optimum conditionsB. napus showed a plating efficiency of 7.8% and shooting efficiency of 17%; forB. oleracea the figures were 2% and 56%, respectively.Abbreviations BAP 6-benzylaminopurine - NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid     

植物蛋白质组学研究进展

植物结构蛋白质组学研究目的在不断变化。早期旨在比较不同基因型和株系,估测系统发育距离。随后是用蛋白N端Edman测序法或氨基酸分析法鉴定蛋白。如今,应用双向凝胶电泳和质谱法,已能联手成功地解决蛋白分离和鉴定问题。这将有助于对不同突变系与野生型作比较和对不同的表达基因鉴定,应用数据库和作图软件有可能获得功能蛋白构象。