播娘蒿下胚轴高频率再生植株因素的研究

以播娘蒿（Descurainia sophia)下胚轴为外植体,研究影响其分化的若干因素。结果表明：用附加2,4－D的培养基,对下胚轴切段预培养5d可提高分化频率;1.7mg/L AgNO3对分化出芽有很大促进作用,最佳激素组合为2.0mg/L 6-BA 0.2mg/L NAA;黄化苗的下胚轴比绿苗的下胚轴对激素反应更敏感,最高分化频率可达86.05%。再生芽转至1／2MS 0.5mg/L NAA 0.5mg/L IBA的培养基中,生根率100％。     

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。     

花生成熟胚胚轴的植株再生和快繁

利用正交试验筛选出了以花生成熟胚胚轴为外植体获得较高频率的不定芽分化和植株再生的条件,并在此基础上对花生试管苗的快速繁殖进行了研究,研究表明,4d龄实生苗的花生胚轴,接种于MS NAA0.4mg/L TDZ0.2mg/L诱导培养基上培养28d后,转移到MSo可获得丛生的再生苗。将丛生苗分离,接种到P1培养基（MS NAA0.4mg/L BAP0.5mg/L）,小苗长大,把较大的试管苗切成带叶节的茎段,接种于含不同浓度的TDZ和BAP的快繁培养基,结果表明,TDZ0.2mg/L最适宜试管苗的快繁,繁殖系数可达4.8/月,试管苗经IBA诱导生根后,移栽田间,开花结果。     

鳄嘴花的组织培养和快速繁殖

以鳄嘴花[Clinacanthus nutans(Burm.f.)Lindau]幼嫩茎段为外植体,研究不同植物生长调节剂对鳄嘴花茎段腋芽诱导、丛生芽分化、增殖及生根培养的影响。结果表明:茎段在培养基MS+1.0 mg·L~(-1) BAP+0.1mg·L~(-1) NAA上诱导产生腋芽,将腋芽转入培养基MS+1.0 mg·L~(-1) BAP+0.1 mg·L~(-1) NAA诱导分化丛生芽,分化率高达93.3%;在培养基中分别加入30 g·L~(-1)椰汁、30 g·L~(-1)香蕉、0.5 g·L~(-1)蛋白胨,都有壮苗的效果;最佳的生根培养基为MS+0.5 mg·L~(-1) NAA+2.0 mg·L~(-1) IBA,生根率达90%,且根系发达,芽苗生长健壮。移栽至混合基质(泥炭:椰糠:珍珠岩:河沙=3:2:2:1)中,鳄嘴花的成活率达97%。     

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％。     

欧李“大磨盘”丛生芽的诱导及生根技术

以欧李"大磨盘"的茎尖为外植体,1/2MS和MS为基本培养基,研究了不同生长激素对启动培养的影响,不同培养基对丛生芽增殖的影响,初步建立了欧李"大磨盘"丛生芽的诱导及生根技术。适宜"大磨盘"的启动培养的生长素为IBA,其最佳丛生芽的培养基为MS+6-BA1.0mg/L+NAA0.01mg/L+IBA0.2mg/L,最佳分化培养基是MS+6-BA0.5mg/L+NAA0.01mg/L+IBA0.2mg/L,最佳生根培养基是1/2MS+IBA0.5mg/L+NAA0.1mg/L。     

中国木薯栽培种通过器官发生再生植株研究

本文研究了中国木薯栽培种四种外植体通过器官发生再生植株的条件。结果表明:在MS附加0.05mg/L TIBA,1mg/L BA的培养基上“NZ 188”初步的萌发胚状体“切头”后切口处可直接产生丛芽,出芽率为43%。“SC201”胚状体子叶块在MS附加0.5 mg/L NAA,0.5mg/L BA的培养基上可直接出芽,出芽率为42%,在MS附加0.5mg/L IBA,1.5mg/L BA培养基上·出芽率为31%,AgNO_3和ABA单独使用或配合使用均不利于芽的再生。“NZ188”胚状体下胚轴在MS附加0.5mg/LNAA,0.5mg/L BA的培养基上形成的愈伤组织转入MS附加1mg/L NAA,2mg/L BA的培养基上,3周后大多数愈伤组织有绿点出现、仅4.4%外植体分化出芽。“HZ188”无菌苗茎段接种在MS附加0.05mg/L TIBA,2mg/LBA的固体培养基上,2周后形成大量愈伤组织,4周后仅见一块愈伤组织分化出芽。     

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％。     

瓠瓜的离体培养和植株的高效再生

９￣１０ｄ苗龄的瓠瓜无菌苗的带叶基（子叶片与子叶柄之间的部分）子叶在８种分化培养基上培养１５ｄ左右,结果在含３．５ｍｇ／Ｌ ＺＴ的培养基上能以较高的频率直接分化出芽;分化培养前对带叶基子叶及仅含子叶叶基的两种外植体纵向切割都能明显提高出芽率,不含子叶叶基的外植体不能分化出芽,决定出芽的外植体的部位是子叶叶基;纵向切割子叶叶基前预培养９ｄ以上出芽率大幅度提高。     

芥蓝组织培养及再生体系的建立

以黄花芥蓝的子叶、子叶柄和带柄子叶为外植体,在不同的BAP和NAA浓度的组合诱导下,获得再生植株.研究结果表明,在测试激素浓度下,带柄子叶为外植体的再生率最一致,其最适BAP和NAA浓度分别为3.0mg/L和1.0mg/L;较低浓度的BAP和NAA有利于子叶再生,其最适BAP和NAA浓度分别为2.0mg/L和1.0mg/L;子叶柄只有在较高的BAP和NAA浓度下才能再生,但再生率依然很低.     

Micropropagation of Sesbania aculeata (Pers.) by adventitious organogenesis

Multiple shoots differentiated from hypocotyl explants of Sesbania aculeata (Pers.) syn S. cannabina (Retz.) Pers., a leguminous woody shrub, when cultured on Murashige and Skoog's basal medium supplemented with auxin (IBA, NAA) or auxin and cytokinin (IBA + BAP, NAA + BAP). Shoot budding occurred directly from the explant as well as from callus. Differentiation of shoot and root occurred in one step in the same concentration of auxin or auxin and cytokinin. Elongation of shoots occurred in the shoot induction medium.     

In vitro multiplication of Peganum harmala--an important medicinal plant

The frequency of shoot regeneration and multiplication of P. harmala was influenced by the type of explant and kind and concentration of hormones. Of the various seedling explants, cotyledonary node exhibited maximum shoot regeneration frequency from axillary region on MS medium supplemented with 5 microM BAP. Addition of 0.1 microM NAA enhanced the efficacy of BAP for multiple shoot regeneration as well as improved the growth of shoots. BAP (5 microM) in combination with NAA (0.1 microM) was found to be the optimal for inducing an average of 4-5 shoots per explant in 75% of the cultures within 5 weeks. Replacement of BAP with other cytokinins at equimolar concentration of BAP i.e. 5 microM was not effective in inducing multiple shoots. Regenerated shoots were rooted on MS medium containing IBA (8 microM) with 80% efficiency. The plantlets were successfully established in soil where 80% of them developed into morphological normal plants.     

<Emphasis Type="Italic">In vitro</Emphasis> adventitious shoot organogenesis and plant regeneration from seedling explants of <Emphasis Type="Italic">Albizia odoratissima</Emphasis> L.f. (Benth.)

Epicotyl, petiole, and cotyledon explants derived from 14-d-old seedlings of Albizia odoratissima were cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of either 6-benzylaminopurine (BAP) solely or in combination with 0.5 μM naphthalene-3-acetic acid (NAA). The percentage of shoot regeneration and the number of shoots regenerated varied significantly depending on the type of explants used, the concentration of plant growth regulators, and the orientation of explants on the culture medium. The best response in terms of the percentage of shoot regeneration was obtained from epicotyls cultured horizontally on MS medium supplemented with 5 μM BAP, whereas the highest number of shoots per responding explant was recorded on medium containing 2.5 μM BAP and 0.5 μM NAA. Successful rooting was achieved by placing the microshoots onto MS medium containing 25 μM indole-3-butyric acid (IBA) for 24 h first, then transferring to the same medium without IBA. Of the various substrates tested, vermiculite was the best for plant acclimatization, as 75% of the plants survived and became established.     

Micropropagation of black pepper (Piper nigrum Linn.) through shoot tip cultures

Summary The morphogenetic potential of shoot tip explants of black pepper (Piper nigrum) was investigated and an effective multiple-shoot propagation method is described. Various combinations of media, growth regulators and sterilization treatments were compared. Problems with establishment in tissue culture sometimes occurred, probably caused by endogenous pathogens associated with tissue exudates. The best establishment and proliferation of shoot tip explants was obtained on MS medium containing 1.5 mg l^–1 BAP alone; subsequent growth and development of lateral branches was best on media containing 1.5 mg l^–1 BAP plus 3.0 mg l^–1 IBA. Adenine sulphate inhibited the number of explants showing regeneration but increased the number of shoot buds per regenerating explant. Shoots were rooted on a 50% strength medium containing 1mg l^–1 NAA.Abbreviations AdSO₄ adenine hemisulphate - BAP 6-benzylaminopurine - IBA indole-3-butyric acid - NAA napthaleneacetic acid     

In vitro Clonal Propagation and Organogenesis in Spilanthes acmella (L) Murray: A Herbal Pesticidal Plant of North-East India

Multiple shoots were regenerated in MS medium using different concentrations of BAP and Kn and different combinations of BAP with IAA, NAA and IBA. Highest multiplication of shoots was obtained with BAP (0.75 mg l^?1) with 28.4 shoots per explant after 60 days of culture. Shoots rooted best on IBA (0.5 mg l^?1), numbering 48.8 per explant. Organogenesis was maximum in callus cultured on MS medium supplemented with BAP (2.0 mg l^?1) and IAA (1.0 mg l^?1).     

Plant regeneration through the direct induction of shoot buds from petiole explants of Jatropha curcas: a biofuel plant

An efficient and reproducible method for the regeneration of Jatropha curcas plants has been developed. The method employed direct induction of shoot buds from petiole explants, without the formation of an intervening callus using a Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (58.35%) and the number of shoot buds per explant (10.10) were observed when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 µM TDZ after 6 weeks. The induced shoot buds were transferred to MS medium containing 10 µM kinetin (Kn), 4.5 µM 6-benzyl aminopurine (BAP) and 5.5 µM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA and indole-3-butyric acid (IBA). MS medium supplemented with 2.25 µM BAP and 8.5 µM IAA was found to be the best combination for shoot elongation and 3.01–3.91 cm elongation was achieved after 6 weeks. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. The orientation (horizontal or vertical) and source (in vitro or in vivo) of explants also significantly influenced plant regeneration. The elongated shoots could be rooted on half-strength MS medium supplemented with 2% sucrose, different concentrations and combinations of IBA, IAA and NAA, and 0.25 mg L⁻¹ activated charcoal. Half-strength MS medium supplemented with 2% sucrose, 15 µM IBA, 5.7 µM IAA, 5.5 µM NAA and 0.25 mg L⁻¹ activated charcoal was found to be the best for promoting rooting. The rooted plants could be established in soil with more than 90% survival.     

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.     

TDZ-induced high frequency plant regeneration through multiple shoot formation in witloof chicory (<Emphasis Type="Italic">Cichorium intybus</Emphasis> L.)

A very efficient and rapid regeneration system via multiple shoot formation was developed for Cichorium intybus L. when leaf explants excised from sterile seedlings were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators. In a comparison of leaf lamina and petiole explants, lamina explants produced over three times more shoots than petiole explants, with a mean of 7.5 shoots compared to 2.4. Of the combinations of KIN/IAA, KIN/NAA, BAP/IAA, or BAP/NAA, 0.5 mg l⁻¹ KIN combined with 0.3 mg l⁻¹ IAA was the most effective, producing a mean of 19.7 shoots per lamina explant while the control treatment involving no plant growth regulators produced no shoots at all. When either cytokinin was used alone, BAP was found nearly twice more successful than KIN. However, the most effective treatment of all was the combination of 0.01 mg l⁻¹ TDZ and 1.0 mg l⁻¹ IAA, producing as many as 35.8 shoots per lamina explant. This rate of shoot regeneration is remarkably higher than those previously reported for C. intybus, most likely due to the highly inductive effect of TDZ, which was tested for the first time in this species. Rooting of the shoots was readily achieved on medium containing different concentrations of IAA or IBA. IAA was more effective than IBA and resulted in the highest frequency of shoots that rooted (100%) and mean number of roots per shoot (4.2) when used at 0.5 mg l⁻¹. Hardening off process resulted in a production of more than 80% healthy plantlets.     

In vitro regeneration of Echinacea purpurea from leaf explants

Efficient plant regeneration was achieved via organogenesis from callus cultures derived from leaf tissue of Echinacea purpurea. Proliferating shoot cultures were obtained by placing leaf explants on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) combinations. MS medium supplemented with BAP (4.44 M) and NAA (0.054 M) was the most effective, providing high shoot regeneration frequencies (100%) associated with a high number of shoots per explant (7.7 shoots/explant). Plantlets were rooted on MS medium alone or in combination with different concentrations of indole-3-butyric acid (IBA), and high rooting and survival was achieved using MS media without plant growth regulators (PGR). All plantlets survived acclimatization producing healthy plants in the greenhouse. This study demonstrated that adventitious shoot regeneration of E. purpurea from leaf explants can be a useful method for the multiplication of this important medicinal plant.     

Somatic embryogenesis and plant regeneration from leaf derived callus of winged bean [Psophocarpus tetragonolobus (L.) DC.]

Somatic embryos were obtained from callus cultures derived from leaf explants of the winged bean, Psophocarpus tetragonolobus (L.) DC. Initiation and development of the somatic embryos occurred with a two-step culture method. Callus cultures initiated on MS medium with NAA and BAP, upon transfer to a new medium with IAA and BAP, produced somatic embryos. Maximum embryogenesis of 60% was obtained on induction medium with 0.5 mg/l NAA plus 1.0 mg/l BAP followed by transfer to a secondary medium with 0.1 mg/l IAA and 2.0 mg/l BAP. Optimal embryo germination and plantlet development was achieved on MS medium with 0.2 mg/l BAP plus 0.1 mg/l IBA. The regenerated plants were successfully transferred to glasshouse conditions.Abbreviations MS Murashige and Skoog (1962) medium - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - BAP 6-benzylaminopurine - KN Kinetin