水花生愈伤组织的诱导及根的分化

在 1/ 2MS培养基上培养出水花生的无菌苗 ,然后将水花生不同器官的外植体接种在不同培养基和添加不同种类激素及其浓度配比的培养基中 ,研究愈伤组织的诱导条件。结果表明 :茎、叶为适宜组培的器官 ,苄基腺嘌呤(BA)、萘乙酸 (NAA)、玉米素 (ZT)、吲哚乙酸 (IAA)的几种配比对水花生茎和叶的愈伤组织诱导均产生良好效应 ,NAA不变 ,随BA浓度上升 ,愈伤组织的诱导率上升。NAA/BA的比值越大越有利于根的分化。     

怀山药微型块茎愈伤组织的诱导形成及高频率再生

对怀山药微型块茎愈伤组织的诱导及高频率再生进行了研究。结果表明 :(1)光下诱导铁棍山药脱分化形成愈伤组织 ,6 -BA2 mg/L NAA2 mg/L 为最佳激素组合。KT2 mg/L 2 ,4 - D2 mg/L 有利于铁棍山药愈伤组织的增殖。附加 2 mg/L NAA能缩短 4 7号山药愈伤组织的诱导时间。KT2 mg/L NAA2 m g/L 有利于 4 7号山药愈伤组织增殖 ;(2 )不同光照条件对愈伤组织的诱导和增殖影响不同。光照是缩短 4 7号山药愈伤组织诱导时间的另一因素。暗培养有利于愈伤组织的增殖 ,对 4 7号山药来说 ,暗培养下诱导率也较高 ;(3)基因型不同 ,愈伤组织类型不同 ,诱导率和不定芽分化率也有差异 ,4 7号山药高于铁棍山药 ;(4 )KT对 4 7号山药愈伤组织分化形成不定芽起主要作用 ,2 ,4 - D2 mg/L KT2 mg/L 为最佳激素组合 ;(5 )光培养有利于不定芽的分化     

黄山药愈伤组织诱导与分化

采用黄山药野生植株作为外植体,试验了不同激素处理对黄山药愈伤组织的诱导、分化影响,结果表明：不同的外植体的诱导率差别较大,叶片的诱导率最高,最高达到85．7％,茎段的诱导率较低,平均诱导率仅10％左右。以叶片作为外植体诱导愈伤组织的最佳培养基配方为MS 2,4-D2．0mg／L 6-BA2．5mg／L;愈伤组织分化生芽的最佳配方为MS BA1．0mg／L NAA0．5mg／L 蔗糖2％ pH6．4;愈伤组织分化生根的最佳配方诱MS BA1．0mg／L NAA0．1mg／L 蔗糖3％ pH6．80。     

玉竹的组织培养与快速繁殖

以玉竹[Polygonatum odoratum (Mill.) Druce]根状茎、叶片和茎段为外植体,于附加不同激素配比的MS培养基中诱导愈伤组织、不定芽和不定根,探讨增殖培养和植株再生的条件.结果表明,叶片和茎段外植体诱导愈伤组织和芽的分化率很低;而根状茎外植体易于培养,有较高的诱导率和增殖倍数,其愈伤组织、不定芽和不定根的诱导率分别可达87%、90%和99%以上.适宜根状茎外植体愈伤组织诱导的培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA,有利于增殖和丛生芽分化的培养基为MS+2.0 mg/L 6-BA+0.5 mg/L IBA和MS+3.0 mg/L 6-BA+0.1 mg/L NAA,而1/2MS+3.0～5.0 mg/L NAA适宜诱导试管苗生根培养.试管苗的移栽成活率可达85%以上.     

连钱草愈伤组织诱导及增殖研究

对连钱草Glechoma longituba (Nakai) Kupr.愈伤组织培养作了初步的探索,以不同的培养条件,利用连钱草的顶芽、叶、叶柄为外植体,研究了连钱草愈伤组织的培养。结果表明：在以MS培养基和LS培养基为基本培养基附加不同外源激素2,4-D、NAA、KT、BA条件下,连钱草在一个较宽的生长范围内,均可诱导产生连钱草的愈伤组织,但不同外植体、不同类型植物激素及其不同浓度对愈伤组织发生均有一定影响：作为外植体,连钱草叶柄和叶都可顺利诱导出愈伤组织;生长素2,4-D对连钱草外植体的脱分化起促进作用,但NAA却抑制愈伤组织的形成;细胞分裂素KT和BA均能与2,4-D组合促进愈伤组织的诱导。MS+2,4-D在光暗交替条件下和LS+2,4-D在黑暗条件下有利于连钱草愈伤组织的诱导,最佳诱导和增殖条件是MS+2,4-D（1.5 mg·L^-1）+BA（1.0 mg·L^-1）光、暗交替（光照14 h·d^-1）。在此条件下, 30 d后,叶的诱导率达91.38%,叶柄的诱导率达100%;愈伤组织继代培养14 d后,平均增殖率达202.2%。     

资源植物罗布麻的愈伤组织诱导和植株再生研究

以罗布麻（Apocynum venetum L.）无菌苗的叶、茎和根作为外植体,在不同激素与浓度组合的MS培养基上进行愈伤组织诱导和植株再生研究。结果表明,在多个激素浓度配比的培养基中,罗布麻叶和茎的愈伤组织诱导率均可达到100%;但进一步将愈伤组织转接到含有不同激素组合的培养基进行不定芽分化时,叶和茎来源的愈伤组织的不定芽分化率有很大不同,茎来源的愈伤组织虽可在多个培养基中有不定芽分化,但最高诱导率仅达20%;而叶片来源的愈伤组织虽仅有4个激素组合培养基中有不定芽的分化,但最高分化率可达到35%。因此,构建罗布麻再生体系的最佳外植体应选择叶片。愈伤组织诱导与不定芽分化的最佳培养基均为：MS＋NAA0.1 mg/L＋6-BA 1.0 mg/L;不定芽在1/2MS＋NAA 0.2 mg/L生根培养基的生根效果最佳,不仅根群质量好,且生根率也高达100%;此再生苗的移栽成活率也最高,在适宜条件下可达75%。     

甘蓝型油菜子叶和下胚轴再生植株无性系建立

以甘蓝型油菜(Brassica napus L.)豫油2号和6257的子叶和下胚轴为材料,在不同激素配比的MS培养基上诱导出了愈伤组织。将经过继代的部分愈伤转入分化培养基,分化结果表明:除基因型、外植体和分化培养基的激素配比对分化率有影响外,诱导愈伤培养基的激素配比对分化率也至关重要。豫油2号的子叶和下胚轴在最适诱导培养基(ZT 1+NAA0.5+2,4-D 0.2 mg/L)和最适分化培养基(ZT4+IAA 0.2 mg/L)组合中的愈伤分化率分别为12.5%和75%;6257的子叶和下胚轴在其最适诱导培养基(KT 2+NAA1+2,4-D 0.2 mg/L)和最适分化培养基(6-BA 4+IAA 0.02 mg/L)组合中的愈伤分化率分别为50%和37.5%。将其最适诱导培养基中的愈伤组织继代达8个月以上,建立了不同继代愈伤的再生植株无性系。     

红叶石楠离体植株再生频率的影响因素研究

以红叶石楠带芽茎段及叶片为外植体,分析激素和培养条件等因子对愈伤组织诱导及植株再生的影响。结果表明,MS+0.10mg/L 2,4-D+0.50mg/L NAA+0.50mg/L 6-BA+0.50mg/L KT为最佳愈伤组织诱导培养基,暗培养的愈伤组织诱导率高于光培养,其愈伤组织诱导率可达100%(带芽茎段)和98%(叶片)。MS+0.50mg/L IBA+2.00mg/L 6-BA+2.00mg/L KT为最佳分化增殖培养基,分化率91%以上,增殖倍数6.8以上,均达到最高。1/2MS+0.50mg/L IBA+0.01mg/L NAA为最佳生根培养基,生根率92%,生根量4.4根/株,均达到最高。     

影响魔芋愈伤组织形成的几个因素

研究了魔芋不同外植体类型、激素种类和激素组合、光暗培养条件、不同生理时期取材等因素对愈伤组织诱导能力的影响以及不同处理所得愈伤组织的分化能力。结果表明 ,不同外植体类型诱导愈伤组织能力顺序为 :顶花芽 >幼嫩芽鞘 >子芋 >根状芋 ;经过 4℃低温预处理的子芋和根状芋比未经预处理的外植体更易诱导出愈伤组织 ;MS +BA1 .0～ 2 .0 +NAA1 .0～ 2 .0培养基均能诱导外植体产生愈伤组织 ,愈伤组织诱导率大多在 5 0 %以上 ,最高达 87.5 % ;在魔芋生长期取材比在休眠期取材更易诱导出愈伤组织 ;不同处理所得愈伤组织在分化能力上具有较明显的差异。     

睫毛萼凤仙花的组织培养及其愈伤组织的超微观察

试验以睫毛萼凤仙花无菌苗的叶片、茎和根为外植体进行离体培养,并对两种具有不同再分化能力的愈伤组织进行电镜超微观察.结果表明:在培养基MS+NAA 0.5 mg/L+6-BA 0.5 mg/L上愈伤组织诱导率达最高,叶片愈伤组织的诱导率为100%,茎的愈伤组织诱导率为67%,而根却没有愈伤组织的发生;叶片愈伤组织在MS+...     

杯山药零余子愈伤组织诱导及植株再生的研究

对怀山药（Ｄｉｏｓｃｏｒｅａ ｏｐｐｏｓｉｔａ）零余子愈伤组织的诱导、分化、再生苗的生根和移栽进行了研究。结果表明：⑴在不同激素组合的培养基上怀山药零余子均能产生愈作组织,而且具有一次成苗的能力。ＢＡ２ｍｇ／Ｌ＋ＮＡＡ２ｍｇ／Ｌ的培养基对诱导愈伤组织最有利,其出愈率达１００％;⑵在愈伤组织的分化中,ＢＡ１ｍｇ／Ｌ＋ＮＡＡ１ｍｇ／Ｌ的激素组合是最佳的,其分化率为６３．６％,且多形成丛生芽;⑶再生植株     

银杏愈伤组织的形成及其中黄酮类化合物的产生

单一激素种类对银杏叶片,叶柄和幼茎愈伤组织的诱导中以NAA的效果最佳,2,4－D次之,6－BA最差,除胚乳外,胚,幼苗的胚根,子叶,幼茎,叶片和叶柄,以及成年树的嫩茎,叶片和叶柄各外植体在本试验条件下都能诱发愈伤组织,其中胚,子叶和叶柄的愈伤组织形成频率均可达到100．0％,叶片和幼茎在光照下的愈伤组织诱导频率比黑暗中的略高,而叶柄和胚根则相反,MS和DCR两种培养基都适合银杏幼苗叶片及叶柄愈伤组织的诱导,两者之间不存显著性差异,测得光照培养的3个组织系（ST1,ST2,ST3)中均含银杏黄酮甙元槲皮素,山柰素和异鼠李素,总含量分别为干重的0．35％,0．29％和0．14％,而黑暗中培养的这3个愈伤组织系则没有银杏黄酮的产生。     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of an endangered plant species, <Emphasis Type="Italic">Thermopsis turcica</Emphasis> (Fabaceae)

This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium supplemented with 0.5–20 μM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin (2–8 μM) alone or in combination with a low level (0.5 μM) of 2,4-D or NAA. The best regeneration of shoots from root explants was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were achieved from cotyledonary explants at 4 μM BA and 0.5 μM NAA. The presence of 0.5 μM or higher levels of 2,4-D in shoot induction medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction.     

银杏愈伤组织培养及其黄酮类化合物的测定(简报)

采用银杏胚、胚乳及3个月苗龄的苗叶为外植体,在附加不同激素的培养基上诱导出愈伤组织,从愈伤组织中提取黄酮类化合物并测定其含量。结果表明,由胚诱导的愈伤组织中黄酮类化合物含量最高。     

In vitro plant regeneration of Aster scaber via somatic embryogenesis

We established an in vitro plant regeneration system via somatic embryogenesis of Aster scaber, an important source of various biologically active phytochemicals. We examined the callus induction and embryogenic capacities of three explants, including leaves, petioles, and roots, on 25 different media containing different combinations of α-naphthalene acetic acid (NAA) and 6-benzyladenine (BA). The optimum concentrations of NAA and BA for the production of embryogenic calli were 5.0 μM and 0.05 μM, respectively. Media containing higher concentrations of auxin and cytokinin (such as 25 μM NAA and 25 μM BA) were suitable for shoot regeneration, especially for leaf-derived calli, which are the most readily available calli and are highly competent. For root induction from regenerated shoots, supplemental auxin and/or cytokinin did not improve rooting, but instead caused unwanted callus induction or retarded growth of regenerated plants. Therefore, plant growth regulator-free medium was preferable for root induction. Normal plants were successfully obtained from calli under the optimized conditions described above. This is the first report of the complete process of in vitro plant regeneration of A. scaber via somatic embryogenesis.     

In vitro plantlet regeneration from cotyledons of the tree-legume Leucaena leucocephala

Two plant regeneration methods applicable to Leucaenaleucocephala were developed. In the first method, involvingorganogenesis via callus formation, cotyledon, hypocotyl and root segments wereinitiated on MS medium containing different concentrations ofN⁶-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), andnaphthaleneacetic acid (NAA). Both compact (type I) and friable (type II) calliwere obtained from the cotyledon and hypocotyl explants treated with differentconcentrations of the growth regulators. Shoots were generated only from thefriable calli formed from the cotyledon explants. The calli formed from thehypocotyl explants did not generate shoots and the root explants died withoutforming callus. Cotyledon explants from 3–4 day old seedlings showedmaximum callus induction compared to those from older seedlings. In a secondmethod involving direct organogenesis, excised cotyledons were cultured on 1/2MS medium containing 10–35 mg l^–1N⁶-benzyladenine (BA) for 7–14 days. Transfer of thecotyledonsto regeneration medium containing low BA resulted in callus formation andsubsequent shoot regeneration from the base of the excised cotyledon explants,with up to 100% frequency. Regenerated shoots rooted best on a basal mediumcontaining no growth regulators.     

Production of glycyrrhizin in callus cultures of licorice

Licorice plants, Glycyrrhiza glabra, G. uralensis, and G. inflata, were investigated for callus induction using Murashige and Skoog (MS) medium combined with auxins and cytokinins. After 4 weeks of culture, 33-100% of leaf or stem explants formed calli. Maximum of shoot induction from callus cultures was achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l alpha-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). These results indicated that all three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l). Glycyrrhizin contents of G. uralensis calli induced using MS medium in combination with NAA and BA [(27.60 +/- 8.47) microg/g DW] or TDZ alone [(36.52 +/- 2.45) microg/ g DW] were higher than those found in other combinations.     

Plant regeneration from different explant types of Bituminaria bituminosa and furanocoumarin content along plant regeneration stages

The first protocol for in vitro plant regeneration from different explants of Bituminaria bituminosa, a pasture and medicinal species, has been established. Three explant types (petiole, leaflet and petiole-leaflet attachment “PLA”) cultured on media with different combinations of benzylaminopurine (BA; 5.0, 10.0 or 20.0 μM) and naphthalene acetic acid (NAA) or indole acetic acid (IAA; 0.5 or 5.0 μM) were tested for calli induction, and with 5 μM BA + 0.5 μM NAA or IAA for shoot development. The average number of shoots (≥5 mm) per callus depended on the explant type and the calli induction medium. The highest average number of shoots per callus was achieved by culturing leaflet and PLA explants on 5 μM IAA + 10 μM BA for calli induction and on 0.5 μM IAA + 5 μM BA for shoot development, and by culturing petiole explants on 0.5 μM NAA + 10 μM BA followed by a second culture on 0.5 μM NAA + 5 μM BA. The highest frequency of shoot rooting was achieved with 10.0 μM NAA and 1.0 μM gibberellic acid (GA₃). Rooted plants were acclimatised in a culture chamber, reaching 96 % survival. Acclimatised plants were transferred to a greenhouse and finally to the field, reaching 100 % survival. The furanocoumarin (FC) accumulation was evaluated in organogenic calli, in vitro shoots, ex vitro plants in the greenhouse and in ex vitro plants in the field (after 1 and 4 months of acclimatisation). The content of FCs depended on the plant material evaluated, being higher in ex vitro plants in the field (up to 9,824 μg g^?1 DW total FC) and lowest in organogenic calli (up to 50 μg g^?1 DW total FC). This effect may be due to cell organization, longer exposure to environmental factors and the developmental stage.     

牛蒡组织培养和高频植株再生的研究

通过对牛蒡(A rctium lapp a L.)不同外植体、不同激素配比的比较研究,建立了牛蒡离体培养高效植株再生体系.牛蒡子叶与下胚轴切段在含2.0 m g/L 2,4-D和0.5~2.0 m g/L BA的M S培养基中愈伤组织诱导率可以达到87%~100%;在1.0~3.0 m g/L NAA和0.5~2.0 m g/L BA的M S培养基上通过愈伤组织间接分化或外植体直接分化形成不定芽,其中愈伤组织分化率可达100%;下胚轴的分化率明显高于子叶,在1.0 m g/L NAA和1.0 m g/L BA的M S培养基上下胚轴直接分化率达77.3%.组织学观察发现牛蒡再生有器官发生和体细胞胚发生两种途径.将生长状态良好的不定芽转至含1.0 m g/L IBA和1.0 m g/L NAA的1/2 M S培养基上生根,移栽,成活率达到93.3%.从诱导愈伤组织到组培苗在珍珠岩中过渡成活,大约需要13周.组培苗次年开花并结实,生长形态特征正常.