广藿香不同外植体离体培养的研究

以广藿香叶片、带节茎、不带节茎及根尖为材料进行离体培养,对影响离体再生的因素进行了研究。结果表明:BA有利于广藿香外植体出芽,浓度以0.1～0.5mg/L效果较好;不同外植体的出芽能力有较大差异,其中以叶片和带节茎出芽能力较强,出芽率均达100%;外植体在培养基上的接种方式对出芽也有一定影响,带节茎以形态学下端垂直插入,可以缩短出芽时间及增加单个外植体出芽的数量。无根苗生根以MT+IBA0.2mg/L培养基为好,苗的生长较为健壮。     

长叶皇冠的组织培养与快速繁殖技术研究

以长叶皇冠短缩茎为外植体进行离体培养,对外植体灭菌方法以及培养基中不同种类和浓度生长调节剂对增殖、生根的影响等进行研究。结果表明:以0.1%的HgCl2为灭菌剂,采用8min+(6～8)min、间歇时间为24h的间歇灭菌法,可获得20%以上的成活无菌外植体;试管苗茎纵切成4份后,接种在1/2MS+6-BA6.0mg·L-1增殖培养基上培养42d,增殖系数可达9.3;在1/2MS+NAA0.2mg·L-1培养基上培养28d后,生根率达100%,平均每株约生根18条;炼苗后移植成活率100%。     

60Coγ射线对广藿香的辐照诱变及再生植株的RAPD分析

探讨60Coγ射线辐照对广藿香外植体离体再生的影响及再生植株的诱变效应.采用60Coγ射线辐照广藿香外植体,考察辐照对外植体离体再生的影响,并观察再生植株的形态变异;同时采用随机扩增多态性DNA（RAPD）技术分析诱变再生植株的变异情况.广藿香外植体的存活率和外植体再生芽的能力,随着辐照剂量的升高而降低;部分再生植株在形态上发生一些变化;RAPD分析表明,4个辐照剂量的外植体再生植株多态性频率分别为39.13%、36.37%、23.09%及33.33%.60Coγ射线辐射会造成外植体存活率和再生芽能力的下降,辐照外植体培养获得的再生植株,在表型性状及分子水平上出现了一定的分离,为将辐照诱变技术应用于广藿香育种提供了依据.     

四数九里香组织培养(简报)

以四数九里香新生带腋芽茎段为外植体,研究取材时期、6-BA浓度及生长素种类对外植体生长的影响,探讨不同浓度活性炭对外植体褐化的影响。结果表明,四数九里香组织培养适宜的取材时期为4月;外植体腋芽诱导率在6-BA浓度为2.5 mg·L-1时表现较高;IBA是较适宜外植体生长的生长素;活性炭浓度为1.5 g·L-1可有效减轻外植体褐化;组织培养适宜的启动培养基为MS+6-BA 2.5 mg·L-1+IBA 0.1 mg·L-1+GA3 0.3 mg·L-1+活性炭1.5 g·L-1。     

银柴胡离体培养与毛状根诱导技术初步研究

以银柴胡茎段为外植体,经消毒获得无菌再生材料后,筛选发根农杆菌介导毛状根诱导产生的最适条件。结果显示:最适的无菌消毒方法为:70%酒精浸5 s,0.1%升汞消毒3 min,获得了银柴胡离体培养材料;以发根农杆菌A4菌株介导的银柴胡毛状根诱导过程中,与叶片和不带腋芽茎段相比,带腋芽茎段为最适转化外植体,用OD600=0.8的菌液侵染茎段15 min,共培养3 d,800 mg/L头孢噻肟钠除菌,其诱导率及诱导密度最高,分别为100%和4.7,为最适诱导条件。研究结果说明在适合条件下,银柴胡带腋芽茎段适于诱导毛状根。     

广藿香叶盘法高效再生体系主要影响因素的研究

广藿香是重要的芳香药用植物,利用基因工程技术对广藿香进行品种改良,需要建立一个高效的广藿香植株再生体系。该研究以广藿香无菌苗叶片为材料,将叶盘外植体分别置于不同条件下培养,观察、统计其再生植株的数量及生长状况。通过研究15~50 d的苗龄、第2~4节上的叶及培养基中2,4-D、NAA、BA和KT的浓度和配比等因素对广藿香叶盘再生植株的影响,在此基础上优化培养条件,建立广藿香高效再生体系。结果表明:广藿香无菌苗的苗龄、叶片在茎上的着生位置以及培养基中的植物生长调节物质浓度和配比都对广藿香的植株再生有显著影响;优化培养条件为以培养30 d的广藿香无菌苗顶芽下第2对展开叶片切割的叶盘为外植体,在含0.1 mg·L-1NAA和0.5 mg·L-1BA的MS培养基中培养28 d,叶盘的不定芽发生频率达到100%,单个叶盘的平均再生芽数为96.5个,经生根培养及温室炼苗,再生植株的移栽成活率达到96%。广藿香叶盘植株再生体系的建立为其基因转化研究及优良品种的快速繁育奠定了基础。     

不同培养条件对南烛离体叶片不定芽再生的影响

以南烛( Vaccinium bracteatum Thunb.)组培苗离体叶片为实验材料,研究了不同培养条件对其不定芽再生状况的影响并筛选出最适培养条件。结果表明：基本培养基类型、生长调节剂种类及质量浓度、琼脂和蔗糖质量浓度、外植体类型、外植体接种方式和暗培养时间均对南烛离体叶片不定芽的出芽时间和再生率、外植体干枯率以及不定芽的生长状况有明显影响。南烛离体叶片不定芽再生的最佳培养条件为：以中脉横切2次的叶片为外植体,以近轴面面向培养基的方式接种;以1/2MS-1/2WPM为基本培养基,添加7.5 g·L-1琼脂、25.0 g·L-1蔗糖、0.5 mg·L-1 TDZ、5.0 mg·L-12ip或4.0 mg·L-1 ZT,暗培养21 d后转至光照度2000 lx、光照时间16 h·d-1的条件下培养。     

芍药组织培养中污染现象的克服

针对芍药在组织培养过程中外植体污染难于控制的问题进行了研究。试验分别于不同的季节取材后对外植体进行不同的表面消毒处理。结果表明：以芍药带芽茎段、顶芽、侧芽为外植体,采用0.1%~0.2% HgCl2消毒8 min,结合母株黄化处理、4℃消毒液+搅拌的表面消毒处理,可降低外植体初期培养污染率;在培养基中添加抗菌素,可明显控制内生菌的污染。     

离体条件下花芽分化研究的概况

本文论述了在离体条件下,外植体生理状态及培养条件对外植体花芽分化的影响,概述了国内外在这方面的工作进展,并对离体条件下花芽分化的研究作了理论上和应用上的评价。     

乙烯与植物离体培养外植体的器官发生

本综述了乙烯与离体培养外植体器官发生之间的关系,以及乙烯生物合成的调节对外植体不定芽,不定根,花芽分化,原生质体培养及花药培养等过程的影响。     

An efficient in vitro regeneration system for Lythrum salicaria

This report describes an efficient plant regeneration system for the medicinal plant Lythrum salicaria via direct adventitious shoot development from leaf and stem explants. Leaf explants were much more responsive to regeneration than stem segments. Of the hormonal combinations tested, those involving thidiazuron (TDZ; 0.1, 0.3 or 0.5 mg dm⁻³) were more effective than the combinations of other hormones and 0.1 mg dm-3 TDZ combined with either indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was the most productive. Rooting was readily achieved when multiple shoots were singled out and cultured on medium containing different auxins. IAA was the most effective on root development in terms of both the number of roots per shoot and the frequency of rooted shoots. More than 90 % of the regenerants survived after hardening for four weeks at gradually decreased air humidity.     

Cells within the nodal region of carnation shoots exhibit a high potential for adventitious shoot formation

Adventitious shoot formation was studied with leaf, stem and axillary bud explants of carnation (Dianthus caryophyllus L.). The shoot regeneration procedures were applicable for a wide range of cultivars and shoot regeneration percentages were high for all explant types. Using axillary bud explants, shoot regeneration efficiency was independent of the size of the bud and of its original position in the plant. In contrast, shoot regeneration from stem and leaf explants was strongly dependent on their original position on the plant. The most distal explants (just below the apex) showed the highest level of shoot regeneration. The adventitious shoot primordia developed at the periphery of the stem segment and at the base of leaf explants. In axillary bud, stem and leaf explants, shoot regeneration originated from node cells, located at the transition area between leaf and stem tissue. Moreover, a gradient in shoot regeneration response was observed, increasing towards the apical meristem.Abbreviations BA benzyladenine - NAA naphthaleneacetic acid     

根癌农杆菌对巴戟天遗传转化的影响因素

以巴戟天带节茎为材料,研究了根癌农杆菌对巴戟天遗传转化的影响因素。结果表明:外植体感染前先进行2 d预培养,对转化有一定促进作用;外植体与农杆菌共培养时间以3 d为宜;乙酰丁香酮能提高转化效率,抗性芽分化率可达18.0％;外植体与农杆菌共培养后延迟4 d选择,抗性芽分化率有所提高;硝酸银能抑制外植体表面农杆菌的生长,提高GUS阳性芽的比例,硝酸银浓度2 mg/L时,GUS阳性芽比例最高(42.9％)。     

Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron

An efficient in vitro method for multiple shoot bud induction and regeneration has been developed in Artemisia annua L. using leaf and stem explants in various concentrations and combinations of plant growth regulators to evaluate the frequency of regeneration. The sources of explants as well as plant growth regulators in the medium were found to influence the multiple shoot induction. The result shows that the stem segment cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg/l thidiazuron (TDZ) gave a perfect shoot formation (100%) and good shoot multiplication (57 shoots/explant) after 2 weeks of culture. Healthy regenerated shoots were elongated and rooted in MS medium without hormones. The artemisinin content in plants regenerated from stem explants using 0.1 mg/l TDZ was (3.36 +/- 0.36) microg/mg dry weight and two-fold higher than that of in vitro grown plants of the same age [(1.73 -/+ 0.23) microg/mg DW]. This system exhibited a potential for a rapid propagation of shoots from the stem explant and makes it possible to develop a clonal propagation of A. annua.     

Age and orientation of the cotyledonary leaf explants determine the efficiency of de novo plant regeneration and Agrobacterium tumefaciens-mediated transformation in Jatropha curcas L.

Effects of age and orientation of the explant on callus induction and de novo shoot regeneration from cotyledonary leaf segments of Jatropha curcas were studied. The callus induction and shoot regeneration capacity of cotyledonary leaf segments were found significantly related to the age of the explants and their orientation in culture medium. The youngest explant, derived from the cotyledonary leaf of germinated seed induced the highest regeneration response as compared to one- and two-week-old explants. A gradient response with age of the explant was observed in percentage of callus induction, shoot regeneration from callus and the number of shoots per regenerating callus. The explants cultured with their abaxial side in medium showed significantly higher regeneration response. The youngest explant was found to be most amenable to Agrobacterium-mediated transformation as compared to older explants. The fact that callus induced from the edges of the explant followed by de novo shoot induction, and strong transient gus expression observed in the edges of the explant are significant for routine Agrobacterium-mediated transformation and generation of stable transgenic plants in J. curcas.     

Formation of shoots from leaf axils of Alstroemeria: The effect of the position on the stem

Direct shoot regeneration was induced from leaf explants of Alstroemeria. The explants contained a leaf blade and a small portion of stem node, which were cut from the erect shoots of in vitro multiplicated plantlets. The shoot regeneration capacity of the excised leaf explants was significantly related to the position of the explant on the stem. The youngest explant which was located closest to the shoot apex gave the highest response. A gradient response toward the shoot apex was observed in percentage of shoot regeneration and in the number of shoots per regenerating explant. Histological studies revealed that the shoots were initiated at the leaf axils. The origin of the adventitious buds was located at the epidermal layer of stem peripheral cells. This revised version was published online in June 2006 with corrections to the Cover Date.     

Hormonal requirement and tissue competency for shoot organogenesis in two cultivars of Brassica napus

An investigation of the regeneration ability of explants taken from the floral stem of Brassica napus var. oleifera was performed in the winter cultivars Darmor and Bienvenu. Our purpose was to compare the regeneration ability of the two genotypes, to compare the competence of the different tissues of the stem and then to study histologically the regeneration of shoots. A strong genotypic effect was observed between the two cvs; Bienvenu had a poorer ability to produce shoots when cultured in the presence of benzyladenine: regeneration commenced later; the percentage of explants producing shoots and the number of shoots per regenerating explant were much lower. The comparison between the regeneration ability of different explants, i.e stem segments, internal stem segments, thin cell layer and peels, showed that the superficial tissues were able to regenerate roots but not shoots. Contrariwise, internal stem segments regenerated only shoots. The origin of shoots was investigated in stem segments of cv. Darmor. A kinetic histological analysis showed the basic role played by phloem and phloem-associated cells in shoot formation.     

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。     

In vitro plant regeneration from leaf explants of Ophiorrhiza japonica

An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm⁻³ BA and 0.2 mg dm⁻³ NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings with age of about 18–27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal MS medium supplemented with 0.5 mg dm⁻³ indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where 100 % survived after 1 month of acclimatization.