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1.
We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog (MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine (BA) and indole-3-butyric acid (IBA). The better medium for shoot multiplication and growth was MS + 5 mg L?1 BA + 0.5 mg L?1 IBA + 20 g L?1 sucrose + 7 g L?1 agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium (macro-elements of MS medium are at half-strength) supplemented with 1 mg L?1 IBA, and the survival percentage was >80 % at 16 weeks after transplanting.  相似文献   

2.
Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.  相似文献   

3.
Season and concentration of sterilizing agents play a significant role for establishment of aseptic in vitro shoot cultures and sprouting of nodal explants from field growing culms of bamboo species. In the present investigation the nodal segment explants of Bambusa tulda Roxb collected in different seasons and treated with various concentrations of HgCl2 showed significant variation in aseptic culture establishment and bud break. The rainy season (July–August) recorded with highest of 78% aseptic culture establishment whereas autumn recorded with lowest 46%. Summer and winter seasons emerged to be the best period, registering > 60% in vitro bud break. On the other hand, the autumn season had the lowest value for bud break, i.e. 42%. Among different doses of sterilizing agent tried, HgCl2 0.1% found to be suitable for maximum aseptic culture establishment (66%) as well as bud break (59%). However, among the interactions, summer season and the dose of 0.1% HgCl2 exhibited maximum of 73% response for both aseptic culture establishment and bud break. MS medium (liquid) enriched with 5.0 µM BA + 5.0 µM Kn [Kinetin (N6-Furfuryladenine)] with additional supplementation of 100 µM glutamine + 0.1 µM IAA supported a maximum in vitro shoot multiplication of 4.75 fold. The proliferated shoots were successfully rooted on MS medium (liquid) supplemented 40 µM coumarin. The plantlets transferred to the polythene bags showed 98% survival.  相似文献   

4.
The objective of the study was to develop an in vitro shoot regeneration protocol by utilising shoot tips explant from Vitex trifolia L. Shoot tip explants obtained from a 3-year old plant was cultured on Murashige and Skoog (MS) medium supplemented with various concentrations (1.0, 2.5, 5.0, 7.5 or 10.0 µM) of thidiazuron (TDZ). The optimal level of TDZ supplementation to the culture medium was 5.0 μM for 15 d induction period. The highest number of shoots (22.2 ± 0.1) and shoot length (5.1 ± 0.1 cm) were achieved when TDZ-exposed explants were sub-cultured on MS medium containing 6-benzyladenine (1.0 µM) and 0.5 µM α-naphthalene acetic acid (NAA) after 8 weeks of culture. In vitro rooting of isolated shoots was achieved best in half-strength MS medium containing 0.5 µM NAA. During the acclimatization period, changes in activities of antioxidant enzymes were observed. Superoxide dismutase activity increased reaching maximum at 28th day after transplantation. Likewise, an upregulation of the catalase, ascorbate peroxidase and glutathione reductase enzyme activities were also observed. These observed changes reflected the ability of plants in developing an antioxidant enzymatic defence system aiding in survival against oxidative stress and in reducing release of free radicals. Plantlets were successfully hardened off and acclimatized in earthen pots containing garden soil with a survival rate of 90 %.  相似文献   

5.
Cerasus humilis is a species of small, perennial, drought-resistant and multipurpose deciduous shrub grown in arid and semi-arid conditions in northern China. In this study, an efficient protocol for the rapid micropropagation of C. humilis has been standardized using stem and/or leaf explants. Direct multiple shoot induction was observed when the stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. The highest shoot induction was obtained when stem explants from adult trees were cultured on MS medium supplemented with 2.0 mg L?1 6-benzyladenine (6-BA) and 0.9 mg L?1 α-naphthaleneacetic acid (NAA). The leaf and stem explants cultured on MS medium with 1.0 mg L?1 6-BA and 0.6 mg L?1 NAA, and 0.5 mg L?1 6-BA and 0.8 mg L?1 NAA, respectively, produced the highest induction frequency of callus. Maximum proliferation of callus was observed on MS medium containing a combination of 0.5 mg L?1 6-BA with 0.6 mg L?1 2,4-dichlorophenoxyacetic acid (2,4-d). Optimal shoots differentiated from callus were obtained on MS medium supplemented with 5.0mg L?1 6-BA and 0.9 mg L?1 NAA. In vitro rooting was achieved on half-strength (1/2) MS medium containing 0.5 mg L?1 NAA. Rooted plantlets were hardened under control conditions and successfully acclimatized under field conditions.  相似文献   

6.
An efficient protocol has been developed for in vitro propagation of Enicostema axillare using shoot tip explants. The shoot tip explants were cultured on MS medium supplemented with various combinations of (BAP, KIN) and (NAA/IAA & IBA) in different concentrations between 0.5 and 2.0 mg/l for multiple shoot bud induction. The highest percent of (98.51 %) was observed at 1.0 mg/l BAP in combination with 0.2 mg/l KIN while maximum number of shoot buds (8.41 shoots/explant) was noticed on MS medium containing 1.0 mg/l BAP and 0.2 mg/l KIN combination. The highest frequency (90.82 %) of multiple shoot bud regeneration was observed at 1.0 mg/l BAP and 0.5 mg/l IBA with 15.12 ± 2.12 shoots/explants. The regenerated multiple shoots were transferred to half-strength MS medium augmented with different concentration of 0.5–2.5 mg/l IBA for rooting. Among the different concentrations of IBA tested, maximum percentage of rooting (100 %) was observed in MS medium augmented with 1.5 mg/l IBA. The rooted plantlets were successfully transferred into plastic cups containing soil and sand in the ratio of 1:1. Subsequently established in the field conditions with 90 % of survival rate. The protocol developed can be utilized for both large scale plant production and conservation of germplasm of this species. The described method can be successfully employed for large-scale multiplication and in vitro conservation as well as production of secondary metabolites of E. axillare.  相似文献   

7.
A reliable in vitro regeneration procedure for Populus tomentosa is a prerequisite for its trait improvement through genetic transformation. We established a systematic protocol for indirect regeneration of P. tomentosa using in vitro petioles of Chinese poplar cultivar ‘fasta-3’. A high frequency of callus induction (>97 %) was obtained from isolated petioles cultured on the modified 1/2MS basal medium supplemented with 0.5 mg/L ZT and 1.0 mg/L NAA, and the tested calli were subsequently plated on 1/2MS basal medium supplemented with 0.25 mg/L BA, 0.25 mg/L ZT, 0.25 mg/L NAA, 0.01 mg/L TDZ, and 0.5 mg/L KT for efficient regeneration of shoots after being cultured for 6 weeks. The regenerated shoots were vigorously rooted on the tested media supplemented with 1.0 mg/L IBA and 0.5 mg/L NAA. These results can facilitate genetic transformation of P. tomentosa for trait improvements in future.  相似文献   

8.
An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and ??-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 ??M, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4?C5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 ??M indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants.  相似文献   

9.
Yin Wang  Ruiling Yao 《林业研究》2017,28(6):1169-1175
A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 μM TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 μM NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites.  相似文献   

10.
Tea tree oil is extracted from the leaves and twigs of Melaleuca alternifolia (Maiden & Betche) Cheel, and it is widely used in medicines, food preservatives, cosmetics and health care products. Traditional propagation of M. alternifolia from seeds does not necessarily transfer the desired characteristics from their mother trees, the seedlings are not uniform, and the multiplication rate from cuttings is relatively low. For these reasons, it is necessary to develop tissue culture techniques for this species. This study showed that an efficient explant initiation medium for M. alternifolia was MS 1/2 + BA 0.6 mg L?1 + NAA 0.1 mg L?1 + sucrose 30 g L?1, which yielded a 75.9 % initiation rate. An efficient multiplication medium was MS + BA 0.3 mg L?1 + NAA 0.15 mg L?1 + sucrose 30 g L?1, which yielded a 4.3 multiplication rate and 3.2 cm shoot length. The rooting medium was MS 1/2 + IBA 0.1–0.25 mg L?1 + sucrose 15 g L?1, which yielded a 100 % rooting rate, 2.94–3.32 roots per individual and 1.36–1.44 cm root length. Local red-core soil was suitable as a transplant medium, and yielded 98 % survival. This study improved the tissue culture technique for mass-propagation of M. alternifolia, enabling the production of high quality plants for market.  相似文献   

11.
A highly reproducible and efficient in vitro shoot regeneration system was developed in a potential medicinal plant, Albizia lebbeck using root explants. Root explants from 15 day-old-aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations (0.5, 2.5, 5.0, 7.5 and 10.0 μM) of 6-Benzyladenine (BA), Kinetin (Kn), 2-Isopentenyl adenine (2-iP) singly as well as in combination with α-Naphthalene acetic acid (NAA) (0.1, 0.5, 1.0, 1.5 and 2.0 μM). The highest rate of shoot multiplication (16.0 ± 1.87 for the average shoot number and 5.16 ± 0.38 cm for shoot length) was achieved on MS medium supplemented with 7.5 μM BA and 0.5 μM NAA. The effects of medium type, medium strength, pH and subculture on shoot induction and proliferation were also tested. An average of 21.6±2.87 shoots per explants could be obtained following this protocol. Rooting was achieved on microshoots using half strength MS medium with 2.0 μM Indole-3-butyric acid (IBA) after four weeks of culture. The in vitro raised healthy plantlets were successfully established in earthen pots containing garden soil and grown in greenhouse with >80% survival rate.  相似文献   

12.
Nitraria sibirica Pall. is a shrub that grows in saline-alkali soil and has traditional medicinal value and potential commercial value. The objectives of this study include induction and multiplication of callus, establishment of a suspension cell line, and isolation of protoplasts from cell suspensions. Murashige and Skoog (MS) medium was used for callus induction from mature seeds of N. sibirica. Seed-derived calluses were further multiplied on MS medium augmented with 0.5 mg L?1 6-benzylaminopurine (6-BA) and 1.0 mg L?1 2,4-dichlorophenoxy (2,4-D) acetic acid. Suspension cultures of N. sibirica were initiated by transferring friable calli to the same liquid multiplication medium. Characterization of the suspension culture was assessed based on fresh mass, dry mass, cell viability and pH value of the culture. A typical growth curve was observed after inoculating 1.5 g of callus in 40 mL liquid medium, including a lag phase, an exponential growth phase, a stationary phase, and a negative acceleration phase. The effect of factors such as pre-plasmolysis, enzyme combination, enzymolysis time and mannitol concentration, on the isolation of cell-derived protoplasts were evaluated to determine the usefulness of suspension cultures. The maximum yield (9.79 × 106 cells/g) and highest viability (79.97%) of protoplast were reached when approximately 1 g of cell suspension (cultured for 6 days) was inoculated for 12 h in cell and protoplast washing solution made of 0.8 mol L?1 mannitol mixture solution, cellulose onozuka R-10 2% (w/v), hemicellulose 0.2%, macerozyme R-10 1%, and pectolyase Y-23 0.5%. Protoplast yield was significantly influenced by pre-plasmolysis and cellulose onozuka R-10 (P < 0.05).  相似文献   

13.
Since the generation of full-sib artificial triploid families, rapid clone establishment and genetic improvements have been needed. Here, we report an in vitro method of direct shoot regeneration of a triploid hybrid poplar [(Populus simonii × P. nigra ‘Italica’) × (P. × ‘popularis’)]. Using different randomized block designs, we selected one triploid to evaluate the explant type, optimal concentrations of plant growth regulators and agar, and culture time under light or dark conditions over 60 days. The highest rate of shoot induction, 80.0%, was obtained using Murashige and Skoog (MS) medium supplemented with 0.2 mg/L benzyladenine, 0.04 mg/L naphthaleneacetic acid (NAA), and 5.5 g/L agar for the first 30 days in the dark, then 3 g/L agar for the next 30 days in light. This last medium yielded the best rate of shoot induction (6.32 shoots/explant). These three media were also used to evaluate the influence of the genotypes of the parents and hybrid triploids on regeneration. Two parents and three of the four full-sib triploids were regenerated successfully; different genotypes and explant types significantly affected the rate of shoot induction and average number of shoots. Leaves but not petioles were a suitable explant. One genotype produced the highest rate of shoot induction of 96.67%. Half-strength MS medium supplemented with 0.2 mg/L indole butyric acid and 0.04 mg/L NAA was the most effective for rooting; rooting rate was 96.67%, survival rate of transplants was 73.33%, and rooting frequency surpassed 85% for each genotype. Overall, this in vitro regeneration system will be useful for the propagation and genetic modification of triploid poplars.  相似文献   

14.
This report describes in vitro shoot induction and plant regeneration from nodal segments of Balanites aegyptiaca on Murashige and Skoog (MS) medium fortified with 6-benzyladenine (BA), thidiazuron (TDZ) and kinetin (Kin) (0.5–20.0 μM). MS medium supplemented with BA (12.5 μM) was the most effective in inducing bud break and growth and also in initiating multiple shoot proliferation. However, the optimal level of TDZ supplementation to the culture medium was 5.0 μM. Shoot cultures were established by repeatedly subculturing the original nodal explants on the same medium. Highest number of shoots (11.5 ± 0.7) and shoot length (5.0 ± 0.2 cm) were achieved when cultures were subcultured on MS medium supplemented with 12.5 μM BA and 1.0 μM α-naphthalene acetic acid (NAA). The shoots regenerated from TDZ supplemented medium when subcultured to hormone free MS basal medium considerably increased the rate of shoot multiplication and shoot length by the end of fifth subculture. Rooting of the shoots was achieved on MS medium augmented with 1.0 μM indole-3-butyric acid (IBA) plus 0.5% activated charcoal followed by their transfer to half strength MS basal medium. The in vitro raised plantlets with well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse with 70% survival rate. The results of this study provide the first successful report on in vitro direct plant regeneration of B. aegyptiaca.  相似文献   

15.
This research reports on an efficient shoot proliferation and callus regeneration system for bamboo.Young, semi-lignified branches with one lateral bud from Drepanostachyum luodianense(Yi et R. S. Wang) Keng f.were used as explants. Disinfection with 0.1% HgCl_2 for 8 min was the optimum treatment and the best medium for bud initiation was Murashige and Skoog(MS) medium containing 3.0 mg L~(-1)6-benzyladenine(BA). Multiple shoots were induced from nodal shoot segments on MS medium containing 5.0 mg L~(-1) BA, 0.5 mg L~(-1) kinetin(Kin), and 1.0 mg L~(-1) naphthaleneacetic acid(NAA). The highest frequency of callus formation(65.6%) was on MS medium containing 4.0 mg L~(-1)2,4-dichlorophenoxyacetic acid(2, 4-D), 0.5 mg L~(-1) NAA, and 0.1 mg L~(-1) thidiazuron(TDZ). The optimum medium for callus proliferation was MS medium with 4 mg L~(-1)2,4-D, 0.5 mg L~(-1) TDZ and 0.5 mg L~(-1) NAA, and the optimum hormone combination was 4 mg L~(-1) BA ? 0.5 mg L~(-1) NAA for callus redifferentiation(up to 85.6%). A 100% rooting was achieved on MS medium supplemented with 2.0 mg L~(-1) NAA and 0.5 mg L~(-1)3-indole butyric acid(IBA). Rooted plantlets were acclimatized in a greenhouse in humus soil ? perlite(1:1) substrate. These micropropagated callus induction and regeneration systems for bamboo will be useful for genetic engineering and multiplication.  相似文献   

16.
巨桉组织培养及工厂化育苗技术的研究   总被引：3，自引：0，他引：3  
巨桉是造纸和人造板的良好材料,其组织培养微繁殖体系已经研究成功。取优树主干基部萌芽或扦插苗1~1.5 cm长的茎段做为外植体,外植体和增殖培养在附加有BAP和NAA的MW培养基上进行。培养条件为26±2℃,日光灯光照10 h.d-1。增殖培养采用BAP 0.1~0.5 mg.L-1和NAA 0.01~0.1 mg.L-1。离体的茎芽适合在B2附加0.5 mg.L-1IBA生根。小苗移栽红心土上,在简易温室成活率高达93%。  相似文献   

17.
Cinchona officinalis (Rubiaceae) is an endemic species of the Loja Valley in southern Ecuador with medicinal uses. Because of over-exploitation in the nineteenth century and more recent disturbances to its ecosystem, C. officinalis populations are threatened. Currently, natural regeneration of the populations is low, despite its high plant regeneration and seed formation capacity. In the present study, an efficient protocol for germination, shoot proliferation and plantlets regeneration was developed for this species. Phenolic content and germination rate of C. officinalis seeds were compared with a control species, C. pubescens. Nodal segments from seedlings of C. officinalis were cultured on Gamborg medium supplemented with different combinations of plant growth regulators. Because the phenol content is high in C. officinalis, the phenolic should be removed with hydrogen peroxide or water washes to stimulate germination. Shoots and callus developed from nodal segments within 45 days using most of the tested combinations of plant growth regulators. The best rates of shoot proliferation, callus formation and adventitious buds were obtained in medium supplemented with 5.0 mg L?1 6-benzyl-aminopurine and 3.0 mg L?1 indole-3-butyric acid.  相似文献   

18.
在鱼木属植物中首次采用根段扦插后萌发的半木质化茎段为外植体，以MS 为基本培养基，对 诱导阶段、增殖阶段、生根阶段等进行研究，得出适宜的诱导培养基为：MS+ BA 0.5 mg · L-1+ NAA 0.1 mg · L-1 + TDZ 0.1 mg · L-1，30 d 的平均诱导率为100%，增殖系数为4.11。增殖阶段适宜的培养基为： MS+ BA 0.5 mg · L-1 + NAA 0.1 mg · L-1。生根阶段：无菌的半木质化苗转接于1/2 MS+ IBA 1.5 mg · L-1 的 培养基培养40 d 后生根率与生根系数分别为92% 与6.3；以泥炭土与细沙（V:V=2:1）为基质，炼苗、移 栽30 d 后，成活率达100%。研究采用的外植体来源丰富、不受季节限制、无菌处理简单，增殖率高且可 进行持续增殖，生根率和生根质量高，为鱼木的规模化栽培所需种苗提供了一种新的有效的繁殖途径。  相似文献   

19.
Nothofagus leoni has a restricted distribution in Chilean forests. This work determines suitable culture conditions forin vitro multiplication and rooting through shoots obtained from seedlings. Broadleaved Tree Medium was suitable for shoot multiplication. A medium with a pulse of 0.55 μM BA in the first subculture and two subcultures on BA-free medium resulted in a multiplication rate at day 63 of × 5.7, without callus growth or shoot neoformation. Rooted shoots of good quality (number and length of roots without callus growth) were obtained with 1.23 μM IBA (91.4% of rooting). The first roots appeared at day 11, reaching a higher speed of rooting at day 15.  相似文献   

20.
This report describes a protocol for the in vitro shoot induction and plant regeneration from epicotyl explants of Cassia angustifolia on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA), Kinetin and 2-iP (0.5–10.0 μM). MS medium supplemented with BA (5.0 μM) was the most effective in inducing adventitious shoots and growth. The highest rate of shoot multiplication was achieved on MS medium supplemented with BA (5.0 μM) and Indole-3-acetic acid (IAA, 1.0 μM). The nodal segments excised from the shoots regenerated from BA (5.0 μM) and IAA (1.0 μM) were used as explants for next three round of micropropagation. The number of shoots significantly increased at successive round of micropropagation. For rooting, MS medium supplemented with 2.0 μM indole-3-butyric acid proved to be better than that supplemented with IAA or α-naphthalene acetic acid. The in vitro raised plantlets with well developed shoot and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse. About 52 plants (85 %) survived out of 60 plants transferred in garden soil.  相似文献   

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