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1.
The effect of Thidiazuron (TDZ), basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba×P. berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba×P. berolinensis. Stems of Populus alba×P. berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1 NAA (naphthaleneacetic acid), and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium (WPM) and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that different concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ (0.1 mg·L-1) resulted in more adventitious shoot regeneration and higher concentration of TDZ (>0.1 mg·L-1) inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.  相似文献   

2.
爬行卫矛下胚轴高频离体再生体系的建立(英文)   总被引：1，自引：0，他引：1  
In this paper,a protocol for efficient shoot regeneration was successfully developed from hypocotyl explants of Euonymus fortunei var.radicans.Some factors that influenced shoot regeneration such as different combinations of plant growth regulators,types of medium and inoculation ways were studied in order to establish an efficient plant regeneration for transformation.The results showed that hypocotyl explants wero horizontally cultured on a basic medium composed of MS medium supplemented with 0.5 mg·L-1 BAP and 0.01 mg·L-1 NAA for induction and development of adventidous shoots.Ninety-four percent of regeneration frequency and 5.1 shoots per explants were obtmned after 30 days of culture.Regenerated shootsproliferated efficiently on a shoot multiplication medium consisting of MS medium containing 1.0 mg·L-1 BAP and 0.1 mg·L-1 NAA.Microshoots were rooted on a rooting medium made up of MS medium enriched with O.5 mg·L-1 IBA and O.5 mg·L-1IAA.After hardening,90% of plants were successfully established under greenhouse conditions.Histological observation revealed that shoot primordium originated from subepidermal cells of hypocotyl explants and directly developed into adventitious shoots without caHus formation.  相似文献   

3.
Axillary buds from 3-yr.-old seedlings of Camptotheca acuminata in the greenhouse were cultured on the different basal media with different concentrations of growth regulators for shoot regeneration for studying the effects of different basal media, different concentrations of growth regulators (BA or TDZ), sucrose, agar and pH value on shoot regeneration from axillary bud. The results showed that B5 and WPM media were the optimal basal media and the optimal phyotohormone was BA of 1.0 mg/L or TDZ of 0.1 mg/L; The concentrations of sucrose of 30g/L and agar of 6g/L were most suitable for the shoot regeneration; pH value from 5.8 to 6.6 were broadly effective, but the best at pH 5.8.  相似文献   

4.
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 5.0μM BA and 0.5μM 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-Dwere 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 0.25μM TDZ and 0.1μM NAA during 30 days for callus induction and then with 5.0μM BA and 0.5μM 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 7.35μM). The highest rooting percentage(35 %) was obtained on medium containing2.46μM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.  相似文献   

5.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Embelia ribes Burm. f., a vulnerable medicinal woody climber of the Western Ghats of India. The in vitro procedure involved three steps that included induction of shoot initials from leaf tissue, regeneration and elongation of shoots from the shoot initials, and rooting of shoots. The induction of shoot initials was achieved on Murashige and Skoog (MS) solid medium supplemented with different concentrations of thidiazuron (TDZ). The best medium for shoot induction was MS with 0.272 μM TDZ. Numerous shoot primordia developed within 2–3 weeks on the leaf margin as well as on the midrib region, without any callus phase. In the second step, the shoot clumps separated from the leaf explant on transfer to MS basal medium, resulting in the differentiation of 90% of the shoot initials into well-developed shoots. The 2- to 3-cm-long shoots rooted on half-strength MS basal medium supplemented with 4.90 μM indole-3-butyric acid (IBA) and 3% (w/v) sucrose in the third stage. The rooted plants could be established in soil with 70% success. This protocol could be utilized for in vitro propagation and conservation of this important threatened medicinal plant.  相似文献   

6.
The purpose of our study was to establish a regeneration system for micropropagation of Populus euphratica Olivier. On the basis of an analysis of plant leaf mineral nutrients, a special medium was proposed, called MP2. In optimizing media for in vitro plant cultures including MS, B5 and MP2 media we employed hormones, auxin IAA, cytokine benzyladenine (BAP) and gibberellic acid (GA) in our factorial experiments on media. Adventitious shoots were derived from cuttings of adult plants taken from Xingjiang, west China, on selected media with MP2 0.5 mg·L-1 BA 0.1 mg·L-1 NAA. The shoots were elongated on a medium with 0.25 mg·L-1 BAP, 0.1 mg·L-1 NAA and 2 mg·L-1 GA and were then rooted on a medium with 0.2-0.5 mg·L-1 IBA. All the media were incorporated with 30 g·L-1 sucrose and an adjusted pH at 6.3.  相似文献   

7.
This paper reports on the effect of various micropropagation factors of Gmelina arborea Roxb. through multiple shoot induction. Factors like the source and age of explants, plant growth regulators （PGRs）, media composition, and carbon source affected multiple shoot-ing in the present study. Among all the explants used, only shoot tips derived from one, two, and three week old seedlings could form multiple shoots. Besides, the formation of multiple shoots depended on the con-centration and combination of PGRs. Among all the PGRs, BAP （6-benzylaminopurine） alone gave the highest regeneration efficiency. Simi-larly, IBA （Indole-3-butyric acid） was the most efficient PGR in inducing root formation in the microshoots. Media composition and carbon source also affected the regeneration efficiency. MS （Murashige and Skoog medium） proved to be the best media for regeneration followed by B5, SH （Schenk and Hilderbrandt medium） and WPM （Woody plant medium） in that order. Similarly, among sugars, only sucrose and glucose sup-ported induction of microshoots. Based on this study we recommend the use of glucose in place of sucrose in MS medium for maximum regenera-tion efficiency.  相似文献   

8.
Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Estab- lishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S. laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7–10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine (BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supple- mented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5°C increased significantly compared with those incubated at 23/20°C directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg·L–1 BA plus 0.2 mg·L–1 NAA. In the presence of 0.2 mg·L–1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization.  相似文献   

9.
A protocol was discussed for high efficient plant regeneration from seven bluegrass (Poa pratensis L.) cultivars via an indirect callus induction and somatic embryogenesis method. Mature seeds were used as explants for callus initiation. Callus induction and proliferation efficiencies were investigated on NB, modified MS (MMS) and MS media, supplemented with 2.0 mg·L-1 2,4-dichlorophenoxyacetic acid (2,4-D). The MMS medium performed best. Based on the MMS medium, direct and indirect callus induction effects of bluegrass from mature seeds were compared at the range of 1-5 mg·L-1 2,4-D contained in the medium. Under the direct callus induction method, the most suitable 2,4-D concentrations varied among cultivars. Under the indirect callus induction method, a significantly high callus induction frequency (93.33%-98.33%) was obtained and there were barely any statistically significant differences among the tested genetically diverse cultivars. Somatic embryos were promoted on the MMS medium supplemented with 3 mg·L-1 2,4-D, 0.1 mg·L-1 kinetin and 0.8 mg·L-1 CuSO4. Embryogenetic calli developed into plantlets on the MMS medium containing different concentrations of thidiazuron (TDZ), and the differentiation frequencies varied in the range from 20.15% to 77.65%. The 0.25 mg·L-1 TDZ was generally the most suitable concentration for the tested cultivars.  相似文献   

10.
We developed a method for in vitro regeneration of Garcinia xanthochymus(yellow mangosteen) from matured seed segments. Multiple shoots were induced on woody plant(WP) medium supplemented with cytokinins.An average of 11 shoots per explant were regenerated from mature seed segments on WP medium containing 20 μM 6-benzylaminopurine. Histological analysis revealed that hypodermal cells of seed segments were initially involved in active division, which later developed into meristemoids, subsequently leading to the formation of shoot buds.Shoot elongation was achieved by repeated subculturing of seed explants in shoot regeneration medium. Rooting of shoots was achieved on WP medium supplemented with indole-3-butyric acid or a-naphthalene acetic acid. Plantlets were transplanted to pots containing soil: compost(1:1) and survival rate was 90%.  相似文献   

11.
In this presentation,we report on de novo and axillary shoot regeneration and rooting of shoots maintained over a long term,from cultures of Tectona grandis L.Shoot-tips of teak shoots forced from epicormic buds were used as the starting material for axenic shoot-culture establishment.Long term maintenance of such axenic shoot cultures was carried out by regular sub-culturing on MS media supplemented with N 6-benzyleadenine (BA,8.8 μmol L-1) and indole-3-butyric acid (IBA,2 μmol L-1) for 24 months.Vigorously growing shoot tips (2 3 cm long) were inoculated on the MS basal medium supplemented with different concentrations (0,1,2,4,6,8 or 10 μmol L-1) of either IBA or α-naphthaleneacetic acid (NAA) for rooting.Axillary and de novo shoots were developed from axillary and cut basal ends of shoots,respectively.Shoots growing on auxins were further sub-cultured (every 15 days) and maintained for 45 days.The greatest number of de novo (5.06) as well as axillary shoots (2.85) was observed on the MS medium supplemented with 10 μmol L-1 NAA or 8 μmol L-1 IBA,respectively,after 45 days.The combinations of both IBA (μmol L-1) + NAA (μmol L-1) were tested at different concentrations (4 + 4,6 + 6,8 + 8) supplemented to a half strength MS basal medium with 0.1% activated charcoal for rooting of decapitated and non-decapitated de novo and axillary shoots.Rooting from non-decapitated de novo shoots was highest (93.33%) with a mean number of roots of 4.61 on this medium,supplemented with 6 μmol L-1 IBA + 6 μmol L-1 NAA,after 36 days of initial culture.Individual auxin,however,was not effective for root induction.Rooted shoots were acclimatized in a green house and after four weeks plantlets were transferred to the field.  相似文献   

12.
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 on1/2MS basal medium supplemented with 0.25 mg/L BA,0.25 mg/L ZT, 0.25 mg/L NAA, 0.01 mg/L TDZ, and0.5 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   

13.
A rapid and efficient method for the regeneration of plantlets from root explants ofRobinia pseudoacacia L. by suspension culture was established. The roots taken from aseptically grown 15-day-old seedlings were used as explants. It was determined that photoperiodicity was necessary for root proliferation, and that the promotive effect of IAA (3-indoleacetic acid) on root proliferation was better than that of IBA (3-indolebutyric acid). The roots cultured in 1/2 MS liquid medium containing 3 μM IAA and 1% sucrose at 25°C under 16-hour photoperiod with 50 μmol m−2s−1 PPFD (photosynthetic photon flux density) shaking at 100 times/min reciprocally showed high efficiency for root proliferation. BAP (6-benzylaminopurine) was found to be essential to induce adventitious shoots from the roots, and the roots cultured in the medium supplied with 3 μM BAP combined with 1–6 μM IAA for 3 weeks under the same conditions as in the root proliferation period were most suitable for adventitious shoot inducement.  相似文献   

14.
Dormant buds of Larix gmelinii(4-30years old)were cultured in vitro.Axillary budsgrew on the explants,and60％-65％ of the explants’axillary buds,a differentiation rate of 60％ wasobtained on the explants collected from the 30-year-old trees.The maximum number of axillarybuds was 26 in one induction per initial explant.Bud clusters were separated into individual budsand most of them elongated into shoots.A few roots grew on the shoots.The MS(Murashige andskoog)and SH(Schenk and Hildebrandt)were more efficient media than the WPM.(Woody PlantMedium).The best hormone Combinations for the axillary bud inductions were BA1+NAA0.01 andBA2+NAA0.2(mg/L).The procedure was as follows:(1)Apical buds were explanted on theno—hormone basal agar medium and grown for 1 or 2 weeks;(2)Explants were transferred onto thebud—inducing medium and grown for 2 weeks and then(3)Cultured on the basal medium withouthormones for axillary bud elongation;(4)Bud clusters were separated and cultured continuously toa minimum height of1  相似文献   

15.
Aldehydes play an important role in atmospheric chemistry and plant direct and indirect defense against environmental stresses.In this study,the emissions of saturated C6-C10 aldehydes from Populus simonii × P.pyramidalis ’Opera 8277’ cuttings were examined by using a gas chromatography/mass spectrometry(GC/MS) technique at three levels of light intensity(400,800 and 1 200 μmol·m-2·s-1).A positive correlation between the emissions of these aldehydes and light intensity was found.Moreover,nordi-hydroguaiaretic acid(NDGA),a special inhibitor of lipoxygenase(LOX),significantly inhibited the emissions of C6-C9 aldehydes at three levels of light intensity,but did not influence the emission of decanal(C10).The emissions of C6-C10 aldehydes in NDGA treated poplar cuttings,exhibited the same positive correlation with light intensity.The results indicated that LOX pathway contributes to the emissions of C6-C9 aldehydes,whereas some pathways regulated by light intensity might be a universal mechanism for emissions of C6-C10 aldehydes.  相似文献   

16.
Eucalyptus is very recalcitrant to in vitro culture.In this research, an efficient shoot organogenesis system was developed using 60-day-old plants of Eucalyptus globulus grown in vitro and non-aerated liquid medium to improve shoot proliferation. Cultures were initiated with hypocotyls and leaf segments from plantlets cultivated on semisolid 1/2 MS modified medium supplemented with 4.44 μM 6-Benzyladenine(BA) and 16.1 μM 1-Naphthaleneacetic acid(NAA). Calli were transferred to shoot induction medium, with either 0.5 or 2.7 μM NAA. Shoot multiplication was carried out on 4.44 μM BA + 0.5 μM NAA medium, and semisolid and non-aerated liquid systems were compared for improving shoot proliferation.Rooting of adventitious shoots was evaluated on medium containing NAA or Indole-3-butyric acid-IBA(5 and16 μM). Callogenesis was obtained from both types of explants, although shoot formation was only obtained from leaf-derived calli. Shoot proliferation on 4.44 μM BA+0.5 μM NAA resulted in the most shoots/callus.Non-aerated liquid medium was more efficient in promoting shoot multiplication(53.5 shoots/callus) than was semisolid medium(28.5 shoots/callus). Levels of phenolic compounds were significantly reduced in the shoots cultivated in liquid medium. Efficient rooting(76%) was obtained using 16 μM IBA.  相似文献   

17.
The present study reports a protocol for germination of Strychnos potatorum (ver. Tel. Chilla) using zygotic embryo culture as an embryo rescue method. A 100% germination rate was obtained by culturing the embryos on full-strength Murashige and Skoog’s medium (MS) containing 20 g/L sucrose in comparison to McCown and Lloyd’s Woody Plant Medium (WPM). Germination rates decreased when the sucrose concentration was lower or higher than 20 g·L-1 . WPM/MS medium containing glucose at levels 30, 20, 15 g·L-1 showed a smaller percentage of germination and at quarter strength, WPM/MS medium with glucose did not respond. Multiple shoot formation was found at 1.0 2.0 mg/L BAP; 3.0 mg/L Kn; 2.0 mg/L TDZ on MS medium with 20 g·L-1 sucrose. Germination rates improved when the embryos were placed upright (vertically) in the medium. The in vitro germinated seedlings were acclimatized in a walk-in-chamber and maintained in the green house with the survival rate of 65% 75%. These plants were transferred to the field and were found to be phenotypically normal, healthy and similar to donor plants. This protocol will be useful to overcome seed dormancy and for rapid multiplication and conservation of S. potatorum using zygotic embryo culture.  相似文献   

18.
Acacia karroo Hayne is an arbor species widely distributed in South Africa with the characteristics of fast growth and drought resistance. The species was introduced to China recently. In vitro culture is an effective method to rapidly produce plants and a strategy to minimize somaclonal variation among regenerated plants. Browning, however, is a problem in establishing the in vitro culture system. The present study diminished the problem by selecting explants, using different browning inhibitors and chilling treatment. Results showed that the use of embryos as explants reduced the browning ratio to 46.7%, whilst stem segment explants were browned up to 56.7%. The adventitious buds were successfully induced in the modified tissue culture medium supplemented with 5.0 mg·L-1 6-BA and 0.1 mg·L-1 NAA. The proliferation coefficient of adventitious buds is 2.8.  相似文献   

19.
The effects of Timentin and cefotaxime (Cef) on shoot regeneration of the London plane tree (Platanus acerifolia Willd.) and their use for the suppression of Agrobacterium tumefaciens in Agrobacterium-mediated genetic transformation were compared. Shoot regeneration was significantly reduced on the media with Cef at concentrations from 100 to 500 mg·L–1. Timentin showed negative effect on plant regeneration at concentrations of 100 and 500 mg·L–1; however, 300 mg·L–1 Timentin was shown to facilitate shoot r...  相似文献   

20.
Eucalyptus adult material requires more successive subcultures in the in vitro multiplication phase for increased vigor and cellular activity. This study evaluated the endophytic manifestation and shoot multiplication of one 13-year-old Eucalyptus benthamii clone under different culture conditions and used canopy branches(CB) and trunk base material as explant sources. The culture media were wood plant medium(WPM), Murashige and Skoog medium(MS) and JADS(Correia and co-authors medium).Based on the results of the initial multiplication experiment, further tests examined sucrose concentrations and p H. Morphophysiology, dry mass production, endophyticmanifestation and histochemical were determined. Explant sources responded differently to MS and JADS media, but the WPM medium promoted homogeneous development.The responses were similar for both explant sources when sucrose concentrations varied. Shoots died in the absence of sucrose, showed high oxidation at 60 g L-1 and optimal development at 30 g L-1. Endophytes were more evident for shoots from the CB origin. Explant sources responded distinctively to treatment due to physiological and intrinsic genetic factors. Therefore, explant sources, different culture media, sucrose concentration and p H may determine micropropagation success and influence the presence and/or intensity of endophytic manifestation.  相似文献   

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