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1.
Protocols are outlined for the regeneration of Curcuma soloensis, an attractive tropical ornamental plant, from young vegetative bud explants. We used both direct and callus-mediated regeneration techniques to produce material suitable for mass propagation and the development of transgenic plants. During direct plantlet propagation, the presence of thidiazuron (TDZ) in the growing medium induced more than three times as many shoots as 6-benzylaminopurine (BA), with a mean of 18.7 shoots per explant on MS medium containing 2.5 μM TDZ compared to 5.0 shoots with 40 μM BA. Subsequently, the shoots rooted readily on MS basal medium that was free of plant growth regulators. During indirect plantlet regeneration, TDZ combined with BA and 2,4-dichlorophenoxyacetic acid (2,4-D) had significant effects on embryogenic callus induction and multiplication. The frequency of callus formation was 91.1% for explants cultured on MS basal medium supplemented with 2.5 μM TDZ, 2.0 μM BA and 1.2 μM 2,4-D. On average 7.1 shoots were produced per callus mass cultured on MS medium supplemented with 2.5 μM TDZ, 9.0 μM BA and 1.2 μM naphthaleneacetic acid (NAA). Regenerated shoots were transferred to MS medium supplemented with 2.5 μM TDZ, to produce multiple shoots. In vitro cultured plantlets readily acclimatized to greenhouse conditions, showing 100% survival rates in a sphagnum, perlite and sand (1:1:1) medium. These plants were transplanted into pots or planted in the field. The ex vitro acclimated plants grew vigorously and produced showy inflorescences 5–6 months after planting. The high-frequency of shoot multiplication and rapid flowering of tissue-cultured plants indicate that C. soloensis has great potential in the floricultural market. 相似文献
2.
狗枣猕猴桃叶片离体培养的器官、体细胞胚形成与植株再生(英文) 总被引：1，自引：0，他引：1
以狗枣猕猴桃试管苗的叶片为外植体,接种于含3%蔗糖和0.2%Gelrite的BW培养基上,外加2,4-D(0,0.1,1和10μmol/L)与玉米素(0,1和10μmol/L)的12种激素组合,置于25℃,光周期为16/8h,光照强度为4000lx的条件下培养。在含1或10μmol/L2,4-D与1或10μmol/L玉米素组合的BW培养基上,产生了体细胞胚,并分化出小植株。随着玉米素浓度的增加,每个外植体上的胚再生频率和体细胞胚的数量也随之增加。同时以叶片为外植体产生的狗枣猕猴桃试管苗的愈伤组织表层产生了不定芽,并抽长成枝。发枝率随着玉米素浓度的增加而增加,并受高浓度的2,4-D所抑制。枝芽转接到含1μmol/LNAA的BW培养基上生根,长成小植株。 相似文献
3.
In vitro shoot regeneration from immature seeds of Epimedium alpinum induced by thidiazuron and CPPU
In vitro propagation of Epimedium alpinum L. was carried out using immature seed explants. The effects of various concentrations of thidiazuron (TDZ) and 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU), on the induction of organogenic callus, were evaluated. Organogenesis occurred most efficiently when explants were transiently exposed (48 h) to 20 μM CPPU or 80 μM TDZ followed by culture on hormone-free woody-plant medium (WPM). Organogenic callus consisting of white, compact clumps of tissue proliferated slowly on hormone-free WPM. To promote adventitious shoot induction, the effects of different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) were investigated. The highest per cent shoot regeneration, 66.7% of explants, and the maximum mean number of shoots, 2.6 per explant, were obtained on WPM containing 1.1 μM 2,4-D and 22 μM BA. Shoots were rooted on hormone-free WPM and well-developed plantlets were successfully transferred to soil. 相似文献
4.
N. Selvaraj A. Vasudevan M. Manickavasagam S. Kasthurirengan A. Ganapathi 《Scientia Horticulturae》2007
Organogenic callus induction and high frequency shoot regeneration were achieved from cotyledon explants of cucumber. About 86.2% of cotyledon explants derived from 5-day-old in vitro raised seedlings produced green, compact nodular organogenic callus in MS medium containing NAA (2.69 μM) and BA (4.44 μM) after two successive transfers at 20 days interval. Adventitious shoots were produced from the organogenic callus when it was transferred to MS medium supplemented with NAA (1.34 μM), BA (8.88 μM), zeatin (0.91 μM) and l-glutamine (136.85 μM) with shoot induction frequency of 75.6%. Shoot proliferation occurred when callus with emerging shoots was transferred in the same medium at an interval of 20 days. Shoots (1.0 cm length) were excised from callus and were elongated in MS medium fortified with GA3 (1.44 μM) and BA (4.44 μM). The elongated shoots were rooted in MS medium supplemented with IBA (3.42 μM) and BA (4.44 μM). Rooted plants were acclimatized in green-house and subsequently established in soil with a survival rate of 80%. This protocol yielded an average of 35 shoots per cotyledon explant in a culture duration of 120–140 days. 相似文献
5.
Shoot cultures were established from nodal explants of 3 pear cultivars by subculture on Murashige and Skoog medium containing mixtures of 3 cytokinins. Zeatin and 6·(γ,γ-dimethylallylamino)purine were both supplied at 10 μM and benzyladenine as follows: ‘William's Bon Chrétien’, 6 μM; ‘Packham's Triumph’, 8 μM; ‘Beurré Bosc’, 10 μM. Production of axillary shoots was greater by basal explants (4–6) comprising the remnants of the previous subculture than by apical explants (2–3) comprising the distal parts of extension shoots. Up to 80% of microcuttings of all cultivars formed roots in vitro with either γ-indolebutyric acid, 10 μM, or α-naphthaleneacetic acid, 10 μM, and approximately 50% of rooted microcuttings were successfully established as container-grown plants. Use of aseptic methods for producing own-rooted pears is discussed in relation to fruit growing and fruit improvement. 相似文献
6.
7.
Micropropagation and accumulation of essential oils in wild sage (Salvia fruticosa Mill.) 总被引：4，自引：0，他引：4
Naser A. Arikat Fawzia M. Jawad Nabila S. Karam Rida A. Shibli 《Scientia Horticulturae》2004,100(1-4):193-202
A protocol for in vitro propagation of the wild three-lobed sage (Salvia fruticosa Mill.) (Synonym, Salvia triloba L.) was developed. Shoot tips were excised from in vitro seedlings and established on MS, Nitch and Nitch (NN), or B5 medium. For shoot proliferation, in vitro nodal and apical explants were cultured on MS medium containing 0.25–2 μM 6-benzylaminopurine (BA), 6-furfurylaminopurine (kinetin), or thidiazuron (TDZ). Proliferated microshoots were rooted on MS medium supplemented with 2.7–11.4 μM indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), or α-naphthaleneacetic acid (NAA). Results indicated that shoots established at 100% regardless of media type, however, shoot height, nodes per shoot, and leaf number were highest for explants established on MS medium compared to NN or B5. Number and height of proliferated shoots, nodes per shoot, and leaf number were highest for nodal explants cultured on a medium containing 0.75 μM BA. Microshoots cultured on a medium supplemented with 2.7 μM IBA exhibited the highest rooting percentage compared to those cultured with IAA or NAA. Essential oil composition in microshoots and shoots of greenhouse-grown plants was determined by gas chromatography/mass spectrometry. The major essential oils detected in both plant materials were α-pinene, 1,8-cineole, camphor, and borneol. No α-thujone or β-thujone was detected. The content of essential oils, camphor, and borneol were higher in the microshoots than in shoots of greenhouse-grown plants. 相似文献
8.
Shoot induction ability of explants of herbaceous peony was investigated in semisolid MS medium containing BA, TDZ and GA3. Callus was readily induced from stem without node and petiole explants within 2 days of culture but failed to generate shoots. Adventitious shoots were successfully produced from meristematic regions only: bud eyes on nodal stem sections, and junctions of petioles and petiolules. No shoots were induced from internode sections, petiole without junctions, or leaf sections. Nodal sections were the most efficient explants. There were up to 20 shoots in one explant generated within 20 days of culture. TDZ was more effective than BA to induce shoots. The 100% shoot induction rate was obtained in medium containing 0.1–3 mg L−1 of TDZ. However, higher concentrations of TDZ inhibited shoot elongation and only large leaf clusters were produced. Combinations of BA and TDZ failed to increase shoot induction rates but caused shoots shorter. The 2–60-min pretreatment of explants with 20 mg L−1 TDZ solution was very effective to induce adventitious shoots directly, but both shoot number and shoot length tended to decrease as treatment time increased. GA3 was beneficial for shoot and stem elongation. 相似文献
9.
秋福红树莓叶片离体再生植株研究 总被引：1，自引：0，他引：1
以红树莓品种秋福(Rubus L.cv.Autumn Bliss)离体叶片为外植体,研究适合其再生植株的叶片部位、放置方式、植物生长调节剂以及暗培养时间等条件。结果表明,叶片外植体接种于MS+TDZ2.00mg/L+IAA0.10mg/L的培养基,暗培养2～3周转移至正常光照下培养效果最好,愈伤组织形成率、不定芽再生率和外植体不定芽数分别为100.00%、95.83%和(5.57±0.27)个。将再生芽接种于1/2MS+IBA0.10mg/L的培养基中,35d后生根率达100.00%。再生植株炼苗后移栽,30d时成活率达97.30%,成功地建立了红树莓叶片的离体再生体系。 相似文献
10.
N. Ahmad 《The Journal of Horticultural Science and Biotechnology》2013,88(4):585-589
SummaryAn efficient in vitro regeneration procedure using thidiazuron (TDZ) has been developed to allow high frequency, multiple shoot induction from cotyledonary node explants of cluster bean (Cyamopsis tetragonoloba). Shoot bud induction occurred on Murashige and Skoog (MS) medium after 4 weeks in the presence of TDZ, followed by transfer onto shoot multiplication and elongation media containing MS salts, B5 vitamins, and different combinations of auxins and cytokinins. Multiple shoots were induced at all levels of TDZ in the medium, but the best proliferation capacity occurred at 5 µM TDZ. Combinations of auxins and cytokinins showed a stimulatory effect on shoot multiplication and also on the length of the newly formed shoots. Maximum shoot induction [i.e., the highest number of shoots (16.0 ± 0.94) per explant] was obtained on agar-solidified medium containing 5 µM benzyladenine (BA) with 0.5 µM indole-3-acetic acid (IAA). Rooting of in vitro-regenerated shoots was achieved in ex vitro conditions by a pulse treatment with 300 µM indole-3-butyric acid (IBA) for 15 min. Rooted plantlets were transferred to soil where 70 – 75% attained sexual maturity and produced viable seeds under greenhouse conditions. The present regeneration system is efficient and can be used in various in vitro manipulation studies. 相似文献
11.
High frequency and direct (without callus) plant regeneration was achieved from whole leaf explants of thornless blackberry (Rubus hybrid) cv. Black Satin (EC No. 381258; PI No. 553272) in vitro. Leaf blade explants from 1-, 3- and 5-month-old mother cultures were cultured on Murashige and Skoog (MS) medium with thidiazuron (TDZ), N6-benzylaminopurine (BAP), indol-3-butyric acid (IBA) and α-naphthalene acetic acid (NAA), alone or in combination. Three-month explants cultured on 0.02 mg l−1 TDZ produced a high regeneration frequency (91.7%) and the most shoots/leaf explant (17.3). The shoot primordia developed within 3 weeks from the point of detachment of the petiole from the leaf blade. The age of the explant source significantly affected the shoot regeneration potential of the leaf explants. Leaves excised from 3-month-old in vitro-cultured shoots performed better than those from 1- and 5-month-old shoots. Shoots rooted best on half-strength MS basal medium with 0.5 mg l−1 IBA and 90% of the plantlets survived acclimatization. The regenerated plantlets were morphologically similar to the mother plants. 相似文献
12.
We investigated in vitro regeneration ability of Prunus microcarpa subsp. tortusa using various explants (root, cotyledon and hypocotyl pieces) and cytokinins [benzyladenine (BA), meta-Topolin (mT) and thidiazuron (TDZ)]. Sectioned cotyledon, root and hypocotyl pieces of in vitro grown seedlings were cultured on Nas and Read Medium (NRM) containing BA (7.5, 10, 12.5, 15 or 17.5 μM), mT (2.5, 5.0, 7.5, 10 or 12.5 μM) or TDZ (2.5, 5.0, 7.5, 10 or 12.5 μM). As a measurement of morphogenetic reaction, the ratios of regenerating explants and the numbers of primary adventitious shoots per regenerating explant were analyzed. Cotyledon explants exhibited higher regeneration ratios than hypocotyl explants, and the root explants were inappropriate for regeneration. Both BA and mT were effective on shoot regeneration but higher regenerating explant ratios were obtained when BA was used. In comparison with BA and mT, the effect of TDZ on enhancing explant regeneration ability was insignificant. Mean number of adventitious shoot per regenerating explant was between 1 and 4, and regenerating explant ratios were between 0% and 77%. The practical appliacations of the results are discussed. 相似文献
13.
H. Yakushiji N. Mase Y. Sato 《The Journal of Horticultural Science and Biotechnology》2013,88(6):874-878
SummaryLeaf fragments of fig (Ficus carica L. ‘Masui Dauphine’) regenerated from in vitro shoot culture were excised and inoculated on MS medium supplemented with different combinations of 2, 4-D, TDZ, and 0.5 mM phloroglucinol. Addition of 2, 4-D induced root formation directly on the explant, and the presence of phloroglucinol significantly increased root formation. When a combination of 2, 4-D and TDZ was added to MS medium containing phloroglucinol, the explants started to produce adventitious buds at the edges. The addition of phloroglucinol was effective in inducing adventitious bud formation. Excised shoots were rooted successfully in MS medium that was either hormone free or supplemented with 1.0 mg l–1 indolebutyric acid. Regenerated plantlets were successfully established in soil after a short period of acclimatization. This is the first protocol of organogenesis and plant regeneration from vegetative organs of Ficus carica L. 相似文献
14.
《Scientia Horticulturae》2002,95(4):319-332
We tested the feasibility of in vitro culture techniques for the propagation of the three endangered cacti species Escobaria minima (Baird) D. Hunt, Mammillaria pectinifera (Ruempler) F.A.C. Weber and Pelecyphora aselliformis Ehrenberg. Twenty-five MS-based proliferation media were tested in preliminary experiments, with different combinations of the auxin NAA and either of the cytokinins BA, kinetin or TDZ. TDZ induced a good proliferation rate, albeit associated with abundant callus formation and hyperhydricity of axillary shoots. A high multiplication rate combined with good quality proliferated shoots and little or no callus induction was observed on media containing BA for E. minima and M. pectinifera and on a medium containing kinetin for P. aselliformis. These results were also confirmed in subsequent experiments in which different explants (shoot tips, bases and longitudinal sections) were used. Micropropagated plantlets were successfully restored to the field, where they reached the flowering stage. Plantlet regeneration of M. pectinifera and P. aselliformis from callus induced on media containing TDZ, but not 2,4-D, was also achieved. 相似文献
15.
Plantlets were obtained from usually dormant axillary buds, excised from the crown of pineapple (Ananas comosus L. Merr.) and grown in culture. Multiple shoots arose from single buds grown on Murashige and Skoog medium supplemented with auxins and kinetin. Shaking culture-flasks during growth increased the number of multiple shoots formed, when compared with stationary liquid cultures. Leaf explants excised from in vitro plantlets developed into a callus capable of plantlet regeneration. Subjecting developing buds to surgical segmentation also resulted in multiple shoot formation. Such shoots, when excised and grown on Murashige and Skoog medium supplemented with auxins, developed roots and grew into complete plantlets capable of being grown in soil. 相似文献
16.
The aim of the present study was to develop a protocol for in vitro propagation of a grape rootstock, deGrasset, characterized by high tolerance to drought and salinity. Four medium compositions, MS, MS with 1/2 nitrates (MS-1), B5 and WPM, were tested for shoot growth from nodal explants and MS-1 medium produced significantly higher rate of shoot proliferation. MS-1 medium supplemented with 2.0 mg/L BAP was found to be optimum for culture establishment. The first subculturing on the same medium resulted in the production of 4–6, mostly short, hyperhydrated shoots per explant. For subsequent subcultures, a reduced concentration of BAP (1.0 mg/L) was used to prevent hyperhydricity, and that resulted in distinct individual shoot elongation. Three plant growth regulators, BAP, ZEA and TDZ, were tested for further shoot proliferation and BAP at 1.0 mg/L added to MS-1 medium displayed the highest proliferation rate (4.75 new explants per explant inoculated, in 6 weeks). For in vitro rooting of shoots, IAA at 0.2 mg/L was found to be optimum to produce highest response (84%) and an average number of 2.03 roots per shoot whereas use of IBA or NAA resulted in rooting with high frequency of callus formation. The acclimatized plantlets were established in soil under net house conditions with 87% success. 相似文献
17.
地被月季‘Royal Bassino’高频再生体系的建立 总被引：7，自引：0，他引：7
以地被月季‘RoyalBassino’为试材, 进行了组培快繁和外植体再生条件的研究。结果表明,以带有腋芽的茎段为外植体, MS中添加6-BA 1.5 mg/L和IBA 0.1 mg/L 为最适诱导培养基, 诱导出芽率100%; MS中添加6-BA 0.5 mg/L和IBA 0.05 mg/L可以获得最高增殖倍数(6.3) ; 而添加IBA 0.01 mg/L的1/2MS为最佳生根培养基, 生根率92%。试管苗经7 d驯化后, 移栽成活率在99%以上。分别以叶柄、叶盘和茎段为外植体进行再生培养, 以MS为基本培养基, 叶柄在添加噻苯隆(TDZ) 7μmol /L的诱导培养基中暗培养10 d后, 转接到添加6-BA 0.5 mg/L的再生培养基中光照培养约20 d, 可以获得57.1%的芽再生率。 相似文献
18.
An adventitious shoot regeneration and rooting protocol was developed for green ash (Fraxinus pennsylvanica) seedling explants. The best regeneration medium for freshly isolated hypocotyls and cotyledons was Murashige and Skoog (MS) supplemented with 13.3 μM 6-benzylaminopurine (BA) plus 4.5 μM thidiazuron (TDZ), and 22.2 μM BA plus 4.5 μM TDZ, respectively. Seventy-six percent of hypocotyl segments and 24% of cotyledon segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 2.7 ± 0.5 and 2.3 ± 1.3, respectively. The effect of in vitro-germinated seedling age on adventitious shoot regeneration from hypocotyl and cotyledon explants was also studied. Results showed that hypocotyl and cotyledon explants from freshly isolated embryos exhibited a higher organogenesis potential than 4–15-day-old explants. Adventitious shoots from hypocotyls and cotyledons were established as proliferating shoot cultures following transfer to MS basal medium with Gamborg B5 vitamins supplemented with 10 μM BA plus 10 μM TDZ. A high rooting percentage (73–90%) was achieved when in vitro shoots were rooted on woody plant medium (WPM) containing 4.9 μM indole-3-butyric acid (IBA) and IAA (0, 2.9, 5.7, or 8.6 μM) with a combination of 10-day dark culture period followed by a 16-h photoperiod. The highest rooting (90%) of adventitious shoots or the number of roots per shoot (3.0 ± 1.0) was obtained on WPM with 4.9 μM IBA plus 5.7 μM IAA. Rooted plants were successfully acclimatized to the greenhouse and 100% survived after overwintering in cold storage. This regeneration system using hypocotyls and cotyledons provides a foundation for Agrobacterium-mediated genetic transformation of F. pennsylvanica for resistance to the emerald ash borer. 相似文献
19.
Adventitious shoots were regenerated from leaf explants of three lowbush blueberry (Vaccinium angustifolium Ait.) clones: ‘QB1’, ‘QB2’ and ‘PB1’ by culturing on a gelled basal medium (BM) with 2.3–4.5 μM thidiazuron (TDZ) for four weeks followed by in a bioreactor system containing the same liquid medium but with 1.2–2.3 μM TDZ for another four weeks. Young expanding basal leaf segments with the adaxial side touching the culture medium and maintained for two weeks in darkness, produced the best results. Callus development and shoot regeneration were genotype dependent. Adventitious shoots were elongated in the liquid BM with 1 μM zeatin and rooted on a three peat: two perlite (v/v) medium. Acclimatized plantlets were grown actively in the greenhouse with an apparent normal leaf and shoot morphology. Ten random ‘QB1’ regenerated plants were screened using 14 expressed sequence tag-polymerase chain reaction (EST-PCR) markers and showed similar monomorphic amplification profiles confirming clonal fidelity of in vitro-derived ‘QB1’ plants. Results obtained suggested the possibility of adventitious shoot regeneration and true-to-type lowbush blueberry micropropagation using a bioreactor system combined with gelled medium. 相似文献
20.
Viable shoot cultures and weaned plants were obtained from cultured apical meristems with 10 Buddleia cultivars giving viabilities of 32–72%. The number of shoots produced, the micropropagation rate and the root number produced in vitro was higher in meristem derived shoots compared to those derived from shoot-tips. The subsequent growth rate of meristem derived plants, in the greenhouse, was also greater. The number of roots produced by conventional cuttings collected from meristem derived plants was significantly higher than in cuttings which were collected from plants derived from shoot-tips or from the original stock plants.Endogenous bacteria were not detected in either shoot cultures derived from meristems or in 10-week-old weaned plants derived from meristems whereas those derived from shoot-tips showed the presence of endogenous bacteria when sterilized explants were cultured on nutrient agar or on tryptic soy broth.Factors affecting adventitious bud and shoot production in leaf and internode explants was determined for ‘Lochinch’, ‘Border Beauty’, ‘Ile de France’ and ‘Pink Delight’ using meristem derived shoot cultures. Adventitious shoots appeared after 4 weeks of culture, in both types of explant when cultured on MS supplemented with 0.5–5.0 μM TDZ. The highest percentage regeneration was achieved from bisected internode explants cultured on 0.5 μM TDZ, with 93–100% regeneration among the cultivars whereas BA was less effective. The best response was obtained using 5.0 μM TDZ which gave over 10–11 shoots per explant in all bisected explants for all cultivars. 相似文献