甘草离体培养物中总黄酮提取工艺的优化

【研究目的】对甘草离体培养物中总黄酮提取和测定工艺进行优化,以建立在室内筛选高产细胞系和试管苗株系的方法;【方法】采用超声波提取技术,对提取液、浸泡时间、提取温度等进行优化;【结果】将样品以75%甲醇浸泡6h后在32℃下超声提取两次,比色法测定总黄酮含量,加样回收率为95.3%～101.4%,相对标准偏差为1.61%。测得甘草初代愈伤组织中总黄酮含量为0.0012%～0.05%,子叶愈伤组织调控后含量高达0.08%,试管苗中总黄酮含量为1.04%～1.31%,移栽后高达4.5%。【结论】此工艺准确度较高、稳定性好、简便可行,为优良细胞系的筛选和试管苗的早期鉴定奠定了技术基础。

植物离体培养物中鱼藤酮的分离与HPLC测定

毛鱼藤（Ｄｅｒｒｉｓ ｅｌｌｉｐｔｉｃａ（Ｒｏｘｂ．） Ｂｅｎｔｈ．）和西非灰毛豆（Ｔｅｐｈｒｏｓｉａ ｖｏｇｅｌｉｉ Ｈｏｏｋ．ｆ．）离体培养物经９：１ＣＨＣｌ３／ＭｅＯＨ溶剂提取，Ｃ－１８柱层析分离后，在室温条件下以６６：３４ＭｅＯＨ／Ｈ２Ｏ或４２：５８ ＣＨ３ＣＮ／Ｈ２Ｏ为流动相，对样品进行反相柱ＨＰＬＣ分析。结果表明，毛鱼藤和西非灰毛豆的愈伤组织均能合成鱼藤酮，但含量相当低，分别为 １７．４ ｕｇ／ｇ，１．８ ｕｇ／ｇ干重。经诱导生根分化的毛鱼藤愈伤组织鱼藤酮含量达１３６．２ ｕｇ／ｇ干重，为愈伤组织含量的７．８倍。西非灰毛豆悬浮培养物的鱼藤酮含量虽比愈伤组织有很大提高，但还不及毛鱼藤愈伤组织含量。结果表明，植物的基因类型在次生代谢上起重要作用，同时离体组织或细胞的分化有助于鱼藤酮的生物合成。

高粱离体培养物主要特征特性的研究分析

对高粱离体培养物的胚性状况、分化出苗方式、辐射处理效应、抗药性等进行了分析。

提高籼稻花药长期继代离体培养物再分化率的研究

对提高长期继代培养的籼稻花药愈伤组织再分化力的方法和技术进行了初步研究。结果表明继代培养基中添加的外源细胞分裂素配比以NAA 3mg/L +KT 1mg/L为好。用山梨醇部分取代蔗糖作碳源可大大提高籼稻花药愈伤组织的分化潜力。籼稻花药愈伤组织再分化前转移到添加ABA的预分化培养基进行预分化培养 ,可充分挖掘继代愈伤组织的再分化潜力 ,使愈伤组织的再分化率大大提高。通过长期继代培养的籼稻花药愈伤组织再生植株的白苗率与成熟胚愈伤组织再生植株白苗率相近。研究结果表明只要培养方法得当 ,籼稻花药愈伤组织经过长期继代培养后仍可保持较高的再分化力 ,因此以籼稻花药愈伤组织作遗传转化受体 ,在技术上是完全可行的。

黄瓜子叶节离体培养物花分化Ca^(2+)敏感期的研究(简报)

植物开花是一个非常复杂的过程,对其机理的研究有着重要的实践意义和理论意义。近百年来进行了广泛的生理生化研究,提出过多种假设。自90年代初成功分离了花器官分化和发育基因以来,使这一研究领域进入了分子水平。钙不仅是植物的矿质营养元素之一,而且Ca2+作为植物的第二信使,参与细胞内多种生理生化活动。许多研究结果表明,Ca2+在成花诱导、花芽形成和分化过程中起重要作用。

荔枝离体培养物DNA提取方法的研究

采用十二烷基硫酸钠(SDS)并补加非水溶性聚乙烯吡咯烷酮法,提取荔枝胚性培养物基因组DNA,有效地去除了植物组织中所含的多酚、单宁、色素及多糖类物质,得到高质量的荔枝胚性培养物基因组DNA。此DNA适合作为PCR模板应用,能较好地用于RAPD分析,这是一种简便、快捷、经济的荔枝胚性培养物基因组DNA提取的方法。

黄瓜子叶培养物花芽分化初期的生理变化

以黄瓜子叶离体培养物直接分化花芽为实验系统 ,研究了其在花芽分化初期的生理变化。结果表明 ,矿质元素含量变化比较复杂 ,培养 2 d时 Cu、Fe水平剧升 ,4 d时 Zn水平剧升 ,6 d时 Ca水平剧降 ;过氧化物酶 (POD)活性总体呈上升趋势 ,可溶性蛋白和核酸含量则呈下降趋势 ;内源多胺中腐胺 (Put)在培养 2 d时达峰值 ,精胺 (Spm)、亚精胺 (Spd)和尸胺 (Cad)则在培养 4 d时达峰值 ;内源激素中生长素 (IAA)和脱落酸 (ABA)在培养 2 d时剧降。

TIB for Micropropagation and the Relationship between Anthocyanins and Chlorophyll of Strawberry Seedlings

In this study, we employed a temporary immersion bioreactor(TIB)system for the micropropagation of strawberry seedlings. The TIB method and the conventional method for the micropropagation of strawberry seedlings were compared in terms of explant propagation coefficient, seedling fresh weight,contents of anthocyanins and chlorophyll, and photosynthetic characteristics. The results showed that an inoculation density of 40 explants/L was suitable for the micropropagation of ’Benihope’ strawberry seedlings in the TIB. The propagation coefficient, fresh weight, photosynthetic rate, stomatal conductance, transpiration coefficient, and the contents of total anthocyanins and chlorophyll of seedlings micropropagated in the TIB were significantly higher than those of the seedlings micropropagated by the conventional method. In conclusion, the TIB method was superior to the conventional method in the micropropagation of strawberry seedlings.The negative reciprocity did not occur between anthocyanins and chlorophyll in strawberry seedlings cultured under forced ventilation such as TIB.

Optimization the Cell Wall Degrading Enzymes and Technique for Isolation of Protoplasts in Potato

Plasma membrane of plant cells is surrounded by cellulose wall and adjacent cells are joined together by a thick pectin rich matrix. Separation of plant cells and removal of the cell wall experimentally, by either a mechanical or an enzymatic process, results in the production ofprotoplast. Protoplasts are useful tools to study the uptake and transport ofmacromolecules and production of somatic hybrids. Protoplasts can be obtained from all types of actively growing young and healthy tissues. The most convenient and widely used source of plant protoplasts is leaf. Juvenile seedling tissues, cotyledons are other alternative tissues most frequently used for protoplasts isolation. All the environmental and genotypic factors, which affect the cell wall thickenings and compactness indirectly, influence the number of protoplasts recovered. Protoplasts are isolated by two methods, mechanical and enzymatic. The enzyme mixture solution of celluiose/macerozyme is used to digest the cell wall. The critical factors affecting the obtaning ofprotoplasts are the kinds of cell wall degrading enzymes, the physiological state of plant leaves, the type of osmotic stabilizers and the composition of reaction solution. With the improvement of technique and enzyme combination rate, the yield of collected protoplasts will be increased higher.

皮肤细胞信号传递

神经酰胺是由神经鞘氨醇和脂肪酸构成的脂肪分子家族，在细胞膜中浓度很高，据认为在皮肤通透性方面起着重要作用。多年来，人们认为神经酰胺纯粹是一类结构成分，但是现在认为，它们还参与细胞的信号传递。在本文中，作者合成了数种神经酰胺衍生物，研究了它们对角化细胞分化的影响。作者发现，有一些衍生物可以明显地提高人正常表皮角化细胞离体培养物中角蛋白1和外皮蛋白的表达，它们还能迅速引起细胞内钙水平的瞬时提高，刺激p42／44胞外信号调节激酶和c-Jun N-端激酶。