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。     

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.     

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.     

藏红花瓣状体的诱导与花柱-柱头状物的定向分化

藏红花 (ＣｒｏｃｕｓｓａｔｉｖｕｓＬ .)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…     

风信子花被外植体年龄对花器官分化的影响

离体培养风信子(Hyacinthus orientalis L．)不同年龄的花被外植体诱导花器官直接再生的实验表明;1．在MS附加6-BA 2 mg／L,2,4-D 0.1 mg／L的培养基上,年龄V的外植体大量衰老,基本丧失器官再生能力,处于年龄段Ⅱ～Ⅳ的外植体可发生玻璃化反应。2．玻璃化反应的外植体转移至MS附加6-BA 0.2 mg／L、NAA 0.005 mg／L的培养基上继续培养30 d后可再生正常的花被片,表明降低培养基中外源激素浓度能够阻止玻璃化反应继续发生。3．在MS附加6-BA 2 mg／L．2,4-D 0.1 mg／L的培养基上,外植体形态学下部可再生雌蕊状早期结构。平均每块外植体分化雌蕊状早期结构数以年龄Ⅲ的外植体最多。     

Role of Exogenous Hormones in Inducing Cells in different Positions of Perianth Explants to Regenerate Flower Buds in Hyacinthus orientalis

Role of the exogenous hormone in inducing different position cells of perianth explants of hyacinth to regenerate flower buds was studied. Experiments showed that (1) Exogenous hormones are necessary for inducing cells of the perianth explant to regenerate the flower buds; (2) Only cytokinine alone could induce the regeneration of the flower buds, the auxin was not necessary; (3) Exogenous hormones in different concentrations could induce cells in the different parts of the perianth explants to differentiate the flower buds: 6-BAP or zeatin 2 mg/L alone could induce cells located at the lower part of the perianth to differentiate flower buds. Combination of 6-BAP or zeatin 2 mg/L and 2, 4-D 0.1 mg/L was advantageous to cells located middle part of the perianth to regenerate the flower buds. Combination of 6-BAP or zeatin 2 mg/L and 2, 4-D 1.0 mg/L could promote cells located at the upper part of the perianth to differentiate flower buds.     

外源激素在诱导风信子花被外植体不同部位细胞再生花芽中的作用

外源激素诱导风信子（Ｈｙａｃｉｎｔｈｕｓ ｏｒｉｅｎｔａｌｉｓＬ．）同一发育时期花被外植体不同部位细胞再生花芽的实验表明∶１． 诱导花被外植体细胞再生花芽,外源激素是必需的;２． 仅有细胞分裂素就可以诱导花芽再生,生长素并不是必需的;３． 花被外植体上的不同部位的细胞再生花芽时,需要不同浓度的外源激素． 单独加６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ可以诱导花被下部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ ０．１ ｍ ｇ／Ｌ的组合有利于花被中部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ １．０ ｍ ｇ／Ｌ的组合能促进花被上部的细胞分化花芽     

Organogenesis and Plantlet Regeneration in Vitro of Populus euphratica

The research of organogenesis and in vitro plantlet regeneration of Populus euphratica Oliver was carried out using the tender shoots from mature tree as initial explants and MS medium as the basic medium. The effects of plant growth regulators (PGR) on the regeneration were compared. The results showed that the concentration of PGR was not strictly required for the organogenesis of the excised organs and callus, but the ratio of BA to NAA was important. Calli could be induced from the excised leaves and stems cultured on the medium with 0.5 mg/L BA and 0.5 mg/L NAA. The embryonic callus could be multiplied in dark on the medium supplemented with 0.25 mg/L BA and 0.5 mg/L NAA. For the adventitious bud regeneration of the leaf and callus, supplement with 0.5 mg/L BA and 0.1 mg/L NAA was appropriate, giving a regeneration frequency of 82.9% and 100%, respectively. The suitable level of BA and NAA for the excised stem's was 0.1 mg/L and 0.01 mg/L respectively, yielding a regeneration frequency of 83 %. Rooting occurred on the MS medium with half strength of macronutrient and addition of 0.015 mg/L NAA, and the rooting rate could reach up to 86.2%. The techniques of somatic cell cloning of P. euphratica was established in vitro. The problems of deterioration of the subcultured shoots were also discussed.     

胡杨离体器官发生及试管无性系的建立

研究了离体条件下胡杨（Ｐｏｐｕｌｕｓ　ｅｕｐｈｒａｔｉｃａ　Ｏｌｉｖｅｒ）茎段、叶片及愈伤组织的器官发生和植株再生技术。离体培养以ＭＳ为基本培养基并附加４０ｍｇ／Ｌ腺嘌呤和５００ｍｇ／Ｌ水解乳蛋白。离体叶片和茎段在ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．５ｍｇ／Ｌ的培养基上诱导产生愈伤组织,并在含０．２５ｍｇ／ＬＢＡ和０．５ｍｇ／ＬＮＡＡ的培养基上继代增殖。ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．１ｍｇ／Ｌ可诱导叶片和愈伤组织发生不定芽,诱导频率分别为１００％和８２．９％,对于茎段,ＢＡ和ＮＡＡ分别为０．１ｍｇ／Ｌ和０．０１ｍｇ／Ｌ时诱导不定芽频率可达８３％。试管苗在大量元素减半并附加０．０１５ｍｇ／ＬＮＡＡ的ＭＳ培养基上诱导生根,生根率达８６．２％。     

魔芋茎尖组织培养和植株再生的研究

以魔芋茎尖、幼芽为外植本,接种于１／２ＭＳ＋１．０ｍｇ／ＬＢＡ＋０．０１ｍｇ／ＬＮＡＡ的培养基中,茎尖、幼芽逐渐生长直接形成幼芽或幼苗;或从茎尖、幼芽由来的膨大的块茎组织表面诱导出幼芽。膨大的块茎组织分割后接种于ＭＳ＋０．０１ｍｇ／Ｌ＋０．０１ｍｇ／ＬＮＡＡ的培养基中进行增殖培养,同时从增殖的块茎组织表面不断地诱导出幼芽。幼芽切块转入不含激素的ＭＳ培养基中,形成幼苗。幼苗切块转入ＭＳ＋０．１ｍｇ／ＬＮＡＡ的生根培养茎中,幼苗生根,形成完整的植株。试管苗移栽６个月后获得干块茎。     

风信子花器官中HAP2基因的分离与表达研究(英文)

在离体条件下,以风信子花被片为外植体,通过控制激素的浓度可诱导花被片、雄蕊或胚珠的再生。近年来,在拟南芥和金鱼草等模式植物中已经分离出了许多控制花器官发育的同源异形基因,如AG,AP1,AP2,AP3等,其中AP2在控制花萼和花瓣形成过程中起重要作用,因此本文从风信子中分离AP2的同源基因,并对它在风信子再生系统中的表达进行了分析。根据AP2同源基因功能域的保守序列设计一对简并引物：5'-TGGGA（A/G）TC（G/T/C）CA（C/T）AT（C/T）TGGA-3'和5'-TCCCA(AGC)(CT)(GT)(AG)CC(AG) CA(CT)TT(AG)TG-3', 以再生的花被片为材料进行RT-PCR,扩增出大小约300bp的片段,序列分析表明该片段的氨基酸序列与AP2同源性高达89%。进而,利用5’和3’Race PCR,得到全长的cDNA。该基因命名为HAP2,GenBank登记号为AF134116,该基因全长1597bp,编码368个氨基酸(Fig.1）。与AP2相比,HAP2也含有10个氨基酸长的碱性功能域,其中KKSR为核定位信号。此外,HAP2也含有两个序列重复的68个氨基酸长的功能域（HAP2-R1,HAP2-R2）,HAP2-R1也含有能形成(-螺旋结构的核心区域,且与AP2-R1中的核心序列100%同源,而HAP2-R2中的核心区域与AP2-R2相比, 缺少9个氨基酸(Fig.2）。RT-PCR结合Southern 杂交结果表明(Fig.3）,HAP     

芥蓝植株再生体系的优化

采用正交设计方法对影响芥蓝植株再生体系的因素进行了优化研究,结果表明：影响芥蓝植株再生的最主要因素是外植体类型,其次依次为NAA,BAP,蔗糖和AgNO3。结果进一步显示,最利于芥蓝再生植株的培养基条件为：MS BA P2mg／L NAA0．03mg／L 1％蔗糖 AgNO3 7．0mg／L 0．8％琼脂,最适宜的外植体类型为下胚轴,植株再生频率高达97．5％。     

白菜高效再生系统的建立及其不定芽发生的组织学观察（简报）

白菜(Brassica campestris Lssp．chinensis Makino)起源于欧洲的野生芸薹,有许多变种和类型,是我国尤其是长江流域及南方各省普遍栽培的重要蔬菜种类之一,在农业生产中占有重要的地位。由于白菜的植株再生频率同其他芸薹属作物相比较低,因此,影响了基因工程技术在白菜品种改良上的应用。虽     

改良菜心离体培养植株再生体系的研究(简报)

菜心(Brassica campestris L.subsp.chinensis Makino var.parachinensis Tsen et Lee)为十字花科芸薹属芸薹种中国白菜亚种中的一个变种,又名“菜薹”。它是我国南方特产蔬菜之一,在蔬菜的周年供应上有重要地位。目前迫切需要培育抗病虫、抗逆和具有其他优良农艺性状的新品种,以提高菜心的     

An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis

In-vitro regeneration in fieldpea was achieved from immature embryonic axes and cotyledonary node explants of six genotypes on modified MS media supplemented with different concentration of plant growth regulators, 6-Benzylamino purine (BAP) and Naphthalene acetic acid (NAA). The best regeneration response, leading to multiple shoot formation efficiency (22.34 shoots/explant) was observed in the medium supplemented with 1.0 mg/L BAP and 0.2 mg/L NAA and best frequency (67.55?±?4.74) was achieved on medium containing 2.0 mg/L BAP and 0.4 mg/L NAA. The shoots were subcultured on a medium supplemented with a combination of 1.0 mg/L GA₃, 2.0 mg/L BAP and 0.4 mg/L NAA, which resulted in elongation of 85 % of shoots. Rooting attempted from the elongated shoots, on half strength MS medium and supplemented with three different auxins IBA, IAA and NAA separately, exhibited similar results. Alternatively, micro-grafting of in vitro regenerated shoots onto pre-germinated root stocks raised in green house facility was attempted with high success rate (75 %). The grafted plants could be successfully hardened, fertigated with Hoagland solution and distilled water in a ratio of (1:10) for acclimatization and further development. All the genotypes tested, produced multiple shoots that could be established to mature fertile plant, hence, the medium combinations used were found to be genotype neutral.     

改良菜心离体培养植株再生体系的研究（简报）

This investigation has developed an efficient and fast method for plant regeneration from petiole of cotyledon explants of Brassica campestris L. subsp. chinensis Makino var. parachinensis Tsen et Lee. A medium was designed for B. campestris subsp. chinensis var. parachinensis to obtain the high frequency of shoot regeneration, which contained BAP 2 mg/L, NAA 0.75-1.0 mgL and 7.5 mg/L AgNO3 solution to the half of NH4+ concentration's MS basic medium. 60 mL/L coconut milk were added to all of media. In this method, frequency of shoot regeneration of "youqing caixin" reached as high as 91.2% and the number of shoots per explant reached as high as 4.7 plants. The result showed that there was a positive correlation between frequency of shoot regeneration and number of shoots per explant. The little shoots could be observed five days after inoculation and were formed directly. The inducing rate of roots of the shoots reached as high as 100% and the rate of viability of transferred mature plant reached higher than 95%. The regeneration period from petiole with cotyledon to a seedling was shorten to about 49 days. Factors influencing in vitro explant regeneration were studied.     

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。     

采用俄罗斯橄榄叶片和茎段诱导产生不定芽的高效再生方法

俄罗斯橄榄(Elaeagnus angustifolia L.)是一种具有很重要药用价值和生态意义的植物。以俄罗斯橄榄一年生幼苗的叶片和茎段为实验材料,探讨了细胞分裂素类(6-BA和Zt)和生长素类(NAA和IBA)两类激素不同组合以及不同配比对植株再生的影响,最后建立了一个高效的俄罗斯橄榄再生方法。结果表明,MS 培养基+ 0.5 mg/L 6-BA +0.2 mg/L NAA更适合叶片的再生,平均每个外植体能产生多达4.3个不定芽;而在MS培养基 + 1.0 mg/L Zt +0.5 mg/L NAA的条件下,茎段外植体再生出来的不定芽最多可以达到平均3.6个;再生芽在含有0.5 mg/L NAA的1/2 MS培养基上生根率达到100%。体外再生苗移栽到装有灭菌混合土(土∶泥炭∶沙子=1∶1∶1)的花盆中锻炼驯化,最后有77%的再生植株存活下来。此结果不仅对俄罗斯橄榄种质资源保护有重要的促进作用,另外也为其将来的遗传转化奠定了基础。     

桃下胚轴高频离体再生体系的建立及优化

以6个桃品种的下胚轴为外植体,研究不同基因型、不同基本培养基及植物生长调节物质组合等对桃下胚轴离体再生不定芽的影响.结果显示:不同基因型桃下胚轴再生率差异显著;QL为'97-8-4'和'甜桃王'的最适基本培养基,MS为'金童5号'的最适基本培养基;植物生长调节物质用2.0 mg/L BA+0.04 mg/L NAA或单独使用2.0 mg/L BA的再生效果好.'97-8-4'在QL+2.0 mg/L BA+0.04 mg/L NAA+100 mg/L肌醇的培养基中再生率最高,为94.44%;'甜桃王' 在QL+2.0 mg/L BA+0.04 mg/L NAA+100 mg/L肌醇+500 mg/L CH的培养基中再生率最高,为80.42%;'金童5号'在QL+2.0 mg/L BA+0.04 mg/L NAA+200 mg/L肌醇的培养基中再生率最高,为78.00%.     

长柄双花木愈伤组织诱导及植株再生

以长柄双花木当年生嫩梢上的叶柄、嫩茎、嫩叶为外植体,对影响长柄双花木愈伤组织诱导和继代、分化主要因素进行研究。结果表明:在培养基MS+NAA0.5mg/L+2,4-D2.0mg/L上,三种外植体均可诱导出愈伤组织,其中叶片愈伤组织诱导率最高。该培养基还可作为愈伤组织继代培养基,但继代培养周期不超过2周。愈伤组织接种在MS+BA2mg/L上分化不定芽,根的诱导在1/2MS+IBA0.5mg/L培养基上进行。