藏红花瓣状体的诱导与花柱-柱头状物的定向分化

藏红花(Crocus sativus L.)又名番红花、西红花;藏药中称为苟日苟木.它是鸢尾科草本植物,原产于欧洲、地中海地区.我国长期以来把它作为珍贵的中藏药.其药用部位是柱头,有效成分主要是藏红花素、藏红花酸、藏红花醛和藏红花苦素.丁葆祖等于1979年首次从藏红花的球茎获得完整的植株[1].Sano等人1987年在离体条件下,诱导花柱-柱头状物再生获得成功[2],此后国内外不断有相关报道[3～7].但试验大多集中在由外植体直接或由愈伤组织间接诱导柱头状物,且花柱-柱头状物的频率低和数量少.我们试图先由藏红花花瓣诱导瓣状体,再定向分化花柱-柱头状物,来提高花柱-柱头状物的诱导频率和数量.从而为以后的大规模生产奠定基础.     

番红花雄蕊柱头状物的离体再生

以番红花（Crocus sativus L.)雄蕊为材料诱导培养出花柱柱状结构,诱导率可达30％,起源于花丝基部。影响雄蕊柱头状物诱导的主要因素为外植体的发育期和生长素NAA的使用浓度。幼嫩浅黄色雄蕊适于诱导柱头状物。温度和光照在不同激素水平下对雄蕊柱头状物诱导的影响不同。紫外检测表明,由雄蕊诱导出的柱头状物含有番红花甙、番红花醛和番红花苦甙。其含量明显高于由药柱诱导出的柱头状结构。在诱导花魔王、子房、花瓣的柱头状物的过程中,观察到成药逆转现象。     

离体培养下番红花花柱—柱头状物再生的研究

番红花(Crocus sativus L.)幼花被和幼花柱可以诱导花柱-柱头状物的再生,长度为24mm 的花芽上分离的花被具有最高的诱导频率(37.5%);花被外植体附加 K5 mg/L 和NAA 4mg/L 有利于再生花柱-柱头状物,频率达到35.3%;附加6-BAP 7 mg/L 和 NAA9mg/L 则有利于柱头状物的大量形成。     

番红花花瓣分步处理与花柱-柱头状物的高频分化

1 植物名称番红花(Crocus sativus),又称藏红花、西红花,藏药中称为苟日苟木. 2 材料类别番红花的花瓣. 3 培养条件第一步诱导瓣状体,诱导培养基为1/2MS+NAA 4 mg*L-1(单位下同)+ KT 4～8+CH 300+肌醇 200.第二步花柱-柱头状物的定向分化培养,分化培养基的配方是1/2MS+NAA 4+KT 4+CH 300+肌醇 200.上述培养基均加入0.5%琼脂粉,3%蔗糖,pH为5.8.培养温度为(20±1)℃,暗中培养.     

花柱和柱头的结构

雌蕊由柱头、花柱和子房３部分组成。柱头是接受花粉的表面,通常将柱头分为２种类型：干砂和湿柱头。干柱头在传粉时没有分泌物的形成,表达干燥并覆盖有一层蛋白质薄膜;湿柱头在传粉时是湿润的,表面被一层分必物所覆盖,有皮细胞人有腺细胞的特征。花柱也被分为２种类型：开放型和闭合型。开放型花柱即中空花柱,花柱中央为花柱道,花柱道周围分布有一层特殊的通道细胞;闭合型花柱即实心花柱,也有２种类型,一类其中央有引导     

中国唇柱苣苔属校订(Ⅰ)

英国学者D.Don于1822年代F.Buchanan发表了Chirita Buch.-Ham.ex D.Don,^[30],于1825年描述了这属的三个种^[13]:C.urticifolia Buch.Ham.ex Don,C.bifolia D.Don及C.pumila D.Don.Chirita属于苦苣苔亚科Cyrtandroideae的长蒴苣苔族Didy-mocarpeae,与这族的另一大属长茹苞苔属Didymccarpus Wall.近缘,二者的主要区别在于柱头的特征上^{[1] [13] [17] [28]},本属的柱头为位于花柱下方的一个薄片,而上方的柱头财完全消失,在长茹筐苔属,柱头呈扁球形或盘状,可能是由二枚柱头合生而成。     

向日葵柱头,花柱和珠孔中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对向日葵（ＨｅｌｉａｎｔｈｕｓａｎｎｕｕｓＬ．）授粉前后柱头、花柱和珠孔中的钙进行了超微细胞化学定位。同时还运用Ｘ射线能谱（ＥＤＸ）和波谱（ＷＤＸ）两种方法进行了Ｘ射线定性分析,证明了前法所得沉淀确系焦锑酸钙。观察表明,花粉萌发和花粉管生长所经的柱头接受面,花柱引导组织和珠孔引导组织中含钙较柱头、花柱和珠孔的其它部位明显地多。柱头乳突细胞的表面和花柱引导组织的胞间基质中、尤其胞间基质与细胞壁外层相接之处钙很密集。在珠孔外端引导组织中,以角质层为界,钙主要分布于其近珠柄侧。花粉管壁果胶质层中有相当多的钙。结合向日葵中已有的研究和其他文献,讨论了钙的分布与花粉管生长的关系     

三种耧斗菜属植物柱头的特征

为了深入了解植物捕获花粉的机制和生殖策略,我们对3种耧斗菜属Aquilegia L.植物华北耧斗菜A. yabeana Kitagawa、无距耧斗菜A. ecalcarata Maxim.和秦岭耧斗菜A. incurvata P. K. Hsiao的雌蕊进行了扫描电镜和荧光显微镜观察,发现其柱头发育的特殊式样。其柱头位于花柱的腹缝面,由花柱腹缝两侧细胞发育成柱头乳突,在花期中从顶端朝花柱基部渐次成熟。伴随花柱向外弯曲,成熟的柱头表面相继呈现,逐渐增加了花粉落置的空间。这种柱头发育的式样通过延长柱头的可授期,增加了植物在不可预测的传粉环境下的繁殖机会。     

番红花

番红花(Crocus sativus)又名西红花、藏红花,为鸢尾科多年生草本。叶线形,自基部鳞茎生出,花顶生,1—2朵,淡紫色,花柱3叉分歧,橙红色至深红色。花期10-11月。番红花原产欧洲南部至伊朗,我国浙江、江苏、江西、山东、西藏、新疆及北京、上海等地有引种栽培。番红花以花柱、柱头入药,能活血化瘀,解郁安神,凉血解毒。番红花     

产藏红花素1(crocin)愈伤组织的诱导及其细胞系的筛选

对藏红花(cfocus sativus L)愈伤组织的诱导条件进行了优化.结果表明:Ms是藏红花芽愈伤组织的最佳诱导培养基,而B5是叶子和花愈伤组织的最佳培养基.藏红花芽、叶和花愈伤组织的最佳诱导温度分别是18℃、25℃和21℃.光照是叶子愈伤组织诱导的有利因素,但不利于芽和花愈伤组织的诱导.1.5～2.0 mg·L-1NAA和0.25 mg·L-6-BA是愈伤组织诱导的最佳激素组合.通过目视法和HPLC方法,从229株细胞系中筛选出细胞系Corml,其藏红花素1的含量是1 677 mg·g-,生长较快,且不易褐化.为采用植物细胞工程法解决藏红花素1资源短缺问题打下了基础.     

In Vitro Regeneration of Style-stigma-like Structure from Stamens of Crocus sativus

Style-stigma-like structures were regenerated from stamens of Crocus sativus L. The age of the stamen explant has an obvious effect on the induction rate. Auxin NAA has larger effect on the induction of filament style-stigma-like structure. Auxin NAA of higher concentration can lead to higher induction rate. Temperature and light have different effects on the induction of style-stigma-like structure from anther's filament of C． sativus with exogenous hormones at different levels. Ultraviolet tests show that style-stigma-like structure from anther's filament of C． sativus contains crocin, safranal and picrocrocin, contents of which are obviously more than those contained in the style-stigma-like from style. Floral reversion was observed in the induction of style-stigma-like structure from petals, ovaries and styles.     

Study on in Vitro Regeneration of Style-Stigma-Like Structure in Crocus sativus L.

Effects of age and variety of explant as well as of exogenous hormones on regeneration of style-stigma-like structure in vitro were observed in saffron. (1) Explants excised from stalk, perianth, anther, ovary, style and stigma showed different reactions in vitro, only young perianth and style could regenerate the style-stigma-like structures; (2) Age of the perianth explant had obvious effect on induction frequency, and the explants excised from an inflorescence of 24mm in length showed the highest frequency (37.5%) and (3) Experiments on exogenous hormones made clear that supplement with K 5mg/L and NAA 4mg/L was advantageous to regeneration of style-stigma-like structure. Induction frequency reached 35.3%. Supplement with 6-BAP 7mg/L and NAA 9mg/L could promote the formation of stigma like structures.     

应用稀土调控藏红花胚性愈伤组织的生长与分化

为提高藏红花（Crocus sativus）胚性愈伤组织的繁殖系数与出芽率,以建立藏红花离体快繁体系,解决其资源短缺问题,采用两步法,用稀土调控其胚性愈伤组织的生长与分化。结果表明,在添加了0.25mg·L-1NAA、3mg·L-16-BA和400mg·L-1CH的B5固体培养基中,0.04mmol·L-1La3＋促进胚性愈伤组织生长的效果最佳,繁殖系数为12,是不添加稀土处理组的1.48倍;在添加了0.25mg·L-1NAA、3mg·L-16-BA和400mg·L-1CH的1/2B5固体培养基中,0.06mmol·L-1Ce3＋促进胚性愈伤组织分化出芽的效果最佳,出芽率高达84.5%,是不添加稀土处理组的1.81倍,且高于国外报道的出芽率（40%）。初步解决了藏红花胚性愈伤组织生长慢和出芽率低等问题,为建立高效稳定的藏红花离体快繁体系奠定了基础。     

木薯离体培养与快速繁殖

选取3个木薯品种的腋芽作为外植体,分别接种于MS基本培养基上进行离体培养及快速繁殖。结果表明, 6-BA不同浓度对木薯品种不定芽诱导效果不一,在MS+6-BA 0.5~1.0mg/L中,3个品种均可诱导产生幼芽;在MS+6-BA 3.0mg/L中,3个品种的幼芽诱导率高达100%;MS+6-BA 3.0mg/L+NAA 0.1mg/L对继代效果较好;MS+NAA 0.1mg/L对生根效果最佳。     

In Vitro Induction of Regeneration of Fruit-Like Structure in Lycopersicon esculentum Mill.

It is possible to induce in vitro regeneration of fruit-like structures from tissue pieces excised from young pistil and young fruit of Lycopersicon esculenturn Mill. These fruit-like structures could be cultured in vitro to maturation, meanwhile they became red coloured. Under dissection it was observed that the fruit-like structure consists of fruit flesh and fruit coats without seeds and plancentae. Tests of exogenous hormones and explant ages revealed: 1. Supplement with exogenous cytokinins alone could induce the explants excised from the pistils to regenerate the fruit-like structures, and the highest induced frequency (50%) was obtained when 0. 5 mg/L zeatin was supplemented. Exogenous auxin seems unnecessary. 2. When tissues of young fruit were used as explants, all explants excised from 4—12 mm (diameter) young fruits could be induced to differentiate the fruit-like structures on medium supplemented with exogenous hormones. However, the hightest frequency of induction (62.5%) was obtained only in tissues of explants excised from 8 mm (diameter) sized fruit and cultured on medium supplemented with 2 mg/L of 6-BAP and 1 mg/L of NAA where regermination of the fruit-like structures took place. In order to investigate the expression of cell totipotency arisen in the regeneration of the fruit-like structure a concept of partial expression of plant cell totipotency was proposed and discussed.     

佳禾早占成熟种胚高效率组织培养方法的研究（简报）

In order to improve the embryogenic callus induction rate and the regeneration rate of JiaHe-ZaoZhan rice, the influence of different factors were investigated, media with different hormones were used. Induction medium was supplemented with 1.5 mg/L 2,4-D + 3 mg/L NAA + 0.1 mg/L KT + 1 mg/L phytic acid + 20 mg/L PAA. Embryogenic call were treated under the condition of 25 degrees C before transferring to regeneration medium, the regeneration medium contained 0.5 mg/L 6-BA + 3 mg/L NAA + 0.5 mg/L KT + 1 mg/L phytic acid. The experiment results indicated that the hormone treatments had certain effects on the callus induction. Under the optimal medium, culture condition and the hormone treatments, the embryogenic callus induction could reach over 95%, and dry treatment of embryogenic callus had been found to increase the frequency of plant regeneration, significantly the plant regeneration rate could reach over 80%. Transplanted into pots, the young plants grew well. Then a experimental system with stability and high regenerating efficiency has been established for the mature seeds of rice (JiaHe-ZaoZhan).     

影响四季桔器官发生的因素

对四季桔胚轴直接出芽、愈伤组织诱导及植株再生能力的研究表明：上、下胚轴在含有BA的MT培养基上,均能直接出芽,但上胚轴出芽能力明显高于下胚轴,最高出芽率为96．9％。外植体在培养基上的接种方式,对出芽也有一定影响,上胚轴切段以形态学下端垂直插入,出芽率高,且在培养前期表现尤为突出。胚轴愈伤诱导的适宜培养基为MT＋IBA0．5mg-1＋BAgmg-1。愈伤组织分化时,上、下胚轴愈伤组织所要求的BA浓度分别为4mgL-1及2mgL-1。在根的诱导中,附加的生长素种类,不仅影响生根率的高低,而且影响上部茎叶的生长,其中以加入lmgL-1IBA的效果最好,生根率达84．4％。     

培养基上生长的黄瓜去根苗雌花高效诱导体系

报告了外源KT(激动素)和IAA(吲哚乙酸)对黄瓜去根苗雌花诱导的增效作用,以及外植体苗龄、下胚轴长度、培养基中N素和NH4 -N水平对成花的影响,据此建立了有效的雌花诱导体系。7d龄带1/2下胚轴的去根苗接种在MS培养基中,对KT3.0mg/L添加IAA0.01mg/L时的雌花成花率达28%,比单用KT3.0mg/L、IAA0.01mg/L的分别高12%、26%,而对照组未见雌花,说明KT和IAA对雌花诱导的增效作用显著。实验表明,N80mmol/L(NH4 -N/TN37.5%)、保留1/4下胚轴和6d苗龄时的黄瓜去根苗雌花诱导率最高。