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1.
A protocol for the production of complete plantlets through multiple shoots from the cotyledon-derived calli of ash gourd (Benincasa hispida L.) is described. The embryos were excised from mature seeds and cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurin (BAP, 1–5 μM). After 10 days the well-developed green cotyledons from the growing embryos were isolated and cultured on MS medium fortified with 2,4-D (1–6 μM). The cultured cotyledons gave rise to luxuriantly growing calli after 6 weeks. These calli were subcultured on MS medium supplemented with various concentrations of BAP (1–6 μM) alone or in combination with naphthalene acetic acid (NAA, 0.2 and 0.5 μM) for regeneration. The regenerated shoots were multiplied and rooted on quarter strength MS medium supplemented with indole-3-butyric acid or NAA (1–5 μM). The rooted shoots were transplanted to soil with 90% success.  相似文献   

2.
A novel micropropagation protocol was established for Capsicum frutescens L. cv. ‘Uchithi’, a pungent chilli cultivar, through induction of axillary shoot proliferation of in vitro raised plantlets by decapitation and using the axillary shoots as explants for multiple shoot bud induction. About 2–6 axillary shoots were induced within 2 weeks when 4-week-old in vitro raised plantlets were decapitated. The axillary shoot-tip explants produced multiple shoot buds when cultured on Murashige and Skoog's (MS) medium containing 8.8–44.4 μM 6-benzylaminopurine (BAP) or 9.3–46.7 μM kinetin alone or 8.8–44.4 μM BAP with 4.6 μM kinetin or 5.7 and 28.5 μM indole-3-acetic acid (IAA). Maximum number of shoots (5.6) were induced on medium containing 22.2 μM BAP in combination with 4.65 μM kinetin. The separated shoots rooted and elongated on medium containing 2.8 μM IAA or 2.4–4.9 μM indole-3-butyric acid (IBA). Rooted plantlets were successfully established in the soil. Efficient mass multiplication of this important food crop was achieved.  相似文献   

3.
The present study was carried out to assess the effect of explant preparation and sizing for in vitro micropropagation of Aloe vera L. The stem nodal explants and shoot tips were cultured on modified Murashige and Skoog's medium (1962) supplemented with different concentrations of 6-benzylaminopurine (BA), kinetin (KIN), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and α-naphthaleneacetic acid (NAA) either singly or in combination. The best media composition was found to be MS medium supplemented with IAA (11.42 μM), IBA (9.8 μM) and BA (8.88 μM). The explants were divided into 2 sets, with and without ensheathing leaf base. Explant sizing, pruning and retention of mother tissue was highly significant in induction of multiple shoots and roots. The stem nodal explants with leaf base performed much better than those without such covering. A very high number of shoots and roots grew from these explants. The rooted plantlets were successfully acclimatized and transferred to the green house conditions and finally to field conditions.  相似文献   

4.
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   

5.
Summary

We have developed a two-step procedure for rooting of tea microshoots in vitro. The effectiveness of different auxin treatments for root formation was found to differ. Among the auxins tested, 25 μM -naphthalene acetic acid (NAA) gave the best results, with 100% rooting, compared to 25 μM indole-3-butyric acid (IBA) or 25 μM indole-3-acetic acid (IAA), which induced 17% and 58% rooting, respectively. Incubation of tea microshoots on 0.33 Murashige and Skoog (MS) medium, supplemented with 25.0 μM NAA or 175.0 μM IBA for 10 d, followed by transfer to auxin-free 0.33 MS medium resulted in 100% rooting, whereas 50.0 μM IAA induced 91.7% rooting. Besides the different auxin treatments, the strength of the MS medium, the duration of incubation of microshoots in auxin-containing medium, the sucrose concentration, the gelling agent, the pH of the medium, the incubation temperature, the light intensity, and the quality of the shoots also played a significant role during in vitro rooting of micropropagated tea shoots. Among the combinations tested, the most effective results were obtained when green microshoots were incubated on 0.33 MS medium supplemented with 25.0 μM NAA, 50.0 mM sucrose, pH 5.5, gelled with 0.2% (w/v) PhytagelTM for 10 d at 25° – 30°C at a light intensity of 40 μmol m–2 s–1, followed by transfer of shoots to auxin-free 0.3 MS medium. This resulted in 100% rooting and, on average, 11 long roots were formed per shoot. Anatomical changes during adventitious rooting of micropropagated tea shoots in vitro were also studied to understand the process of rooting.  相似文献   

6.
In vitro development of isolated embryos and axillary bud proliferation were studied in Pistacia vera L. Different regulator-free nutrient media were compared to determine the effects of the mineral solution, agar and sucrose concentrations on seedling development from mature embryos. Nutrient-rich MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant. 15, 473–479] and DKW [Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19, 507–509] mineral solutions were more efficient for the development of aerial parts than nutrient-poor KN [Knop, W., 1884. Bereitung einer concentrierten nährstofflosung für pflanzen. Landwersuhssat 30, 292–294] and WT [Withe, P.R., 1936. Plant tissue cultures. Bot. Rev. 2, 419–437] solutions. Reducing the agar concentration enhanced fresh matter production and balanced seedling development. When tested at different concentrations, sucrose was found to orient mature embryo development, with the best results obtained at concentrations of 2–4%, whereas high concentrations (6 and 12%) mainly inhibited elongation of the aerial parts. Plantlets obtained previously from in vitro cultured embryos were propagated by axillary budding. High bud proliferation (six shoots per explant) was achieved when using 17.8 μM benzyladenine (BA) combined with 0.5 μM indole-3-butyric acid (IBA). The addition of 2.9 μM gibberellic acid (GA3) to the propagation medium did not improve axillary shoot yields and on average, media with GA3 produced 2.3–2.6 elongated stems per cultured explant. Shoots were rooted in vitro in half-strength MS medium containing 12.3 μM IBA.  相似文献   

7.
《Scientia Horticulturae》2005,105(4):475-482
The present study was conducted to evaluate the regeneration ability of Damask rose. Single-node explants were surface sterilised with 10% chlorox for 15 min and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzyladenine (BA) or kinetin (Kin) separately or in combination with different concentrations of indole-3-butyric acid (IBA). Combination of 2.5–3 mg/l BA and 0.1 mg/l IBA was the most suitable treatment for proliferation. In vitro derived shoots were subcultured four times on the fresh medium within a 4-week period. Other treatments such as explant orientation (horizontal, vertical and oblique) and explant wounding were also examined but did not affect shoot multiplication rate significantly. Several experiments were carried out to stimulate in vitro rooting of Damask rose. Application of different media such as MS, 1/2 MS, 1/3 MS and 1/4 MS with different concentrations of indole-3-acetic acid (IAA), IBA and naphthaleneacetic acid (NAA) did not produce satisfactory results. Quick-dip method using sterilised 0–2000 mg/l IAA, IBA and NAA solutions was also studied. Other treatments such as using various concentrations of abscisic acid (ABA) in combination with various concentrations of IAA, IBA and NAA, and using various concentrations of sucrose and agar did not produce any roots. The best treatment for rooting of shoots was 2.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for 2 weeks in MS medium and then transferring the explants to MS medium without any growth regulator. Plantlets were acclimatised in a soil mixture consisting of peat moss and sand 1:1 (v/v) and successfully transferred to the greenhouse after 3 weeks.  相似文献   

8.
Micropropagation of Phillyrea latifolia L. a wild species present in Mediterranean coastal areas having drought and salt tolerance was performed using explants from adult plants. Shoots were induced from nodal explants on the Rugini’s initial medium (IM). Then these were proliferated on either Rugini olive medium (OM) or Linsmaier and Skoog (LS) medium, each supplemented with 2.22 μM 6-benzylaminopurine (BA) or 4.56 μM zeatin (Z). Rooting (66.1±11%) was induced on shoots grown in perlite soaked with half-strength Rugini olive proliferation medium (OMr) containing 2.69 μM α-naphthaleneacetic acid (NAA) and 160 mg l−1 putrescine. Both shoot multiplication and rooting were performed using Magenta® GA-7 (Sigma) vessels either non-permeable or permeable to gas exchanges. Contamination (about 40%) was observed during the first five passages notwithstanding the addition of cefotaxime to the culture medium, but a high proliferation rate (90%) of explants provided enough healthy plant material. The highest shoot proliferation was observed on LS medium and zeatin whereas the presence of the ventilated filters reduced fresh weight of explants growing on LS media and did not affect shoot growth on OM media. During rooting, the use of ventilated vessels in comparison with the closed ones enhanced development of roots, and doubled the dry weight of plantlets. The vessel ventilation combined with the artificial substrate (perlite) was beneficial for in vitro acclimatization of rooted Phillyrea plantlets.  相似文献   

9.
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   

10.
Nolina recurvata Hemsl. is a very decorative indoor plant but difficult to micropropagate vegetatively. In vitro cuttings failed to induce adventitious rooting. Investigations for a rapid micropropagation using β-cyclodextrin added to the rooting medium has solved the problem. Rooted N. recurvata plantlets were obtained after successive stages of various culture media.Initiation and in vitro multiplication of this plant was possible with lateral buds cultured in Murashige and Skoog medium supplemented with 4.45 μmol of BA and 0.5 μmol of IBA. The number of axillary shoots by explant obtained was 6.In vitro rooting was obtained in MS medium (1/2 strength of minerals salts) supplemented with β-cyclodextrin. This substance, at 1.76 mmol associated with 5 μmol of IBA, improved quality and the rooting rate by 100%.  相似文献   

11.
《Scientia Horticulturae》2005,103(3):381-385
Cornus florida L. (flowering dogwood) has been successfully micropropagated, but low rooting of microshoots makes the system inefficient. This study was conducted to increase rooting efficiency of flowering dogwood microshoots over that previously achieved. Microshoots originating from acclimatized axillary and nodal bud stock cultures were excised and treated with different concentrations and combinations of various auxins including indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA). Effect of microshoot age on rooting efficiency was also examined. Of the auxins tested, maximum rooting was observed with 4.4 μM IBA. The age of microshoot explants had a significant effect on rooting. Five to seven-week-old microshoots treated continuously with 4.9 μM IBA in Woody plant medium (WPM) consistently had the best and most consistent rooting efficiency (about 83%).  相似文献   

12.
RAPD and microsatellite markers were used to determine the genetic fidelity of micropropagated plants from the three varieties of tea plant derived from explants of field grown mother bush as well as in vitro germinated seedlings. Rate of shoot multiplication was better from nodal explants than from shoot tips. A maximum of 32–33 shoots was observed in cotyledonary node in 1/2 MS medium with BAP (6 mg/l), GA3 (1 mg/l) and IBA (0.5 mg/l). 90% of the in vitro derived microshoots were micrografted into rootstocks. The micropropagated plantlets showed both cytological and genetical stability. SSR primers showed complete stability among the regenerants. The results convinced that plants derived from axillary as well as adventitious mode of propagation can be genetically true to type. This cost effective technique would help in fast clonal propagation at a commercial scale.  相似文献   

13.
In vitro obtained shoots of pineapple (Ananas comosus L. Merr.) cv Smooth cayenne was cultured on MS medium enriched with 6-benzylaminopurine (BAP) at 3.25 mg l−1 (14.43 μM) and indole acetic acid (IAA) at 1.75 mg 1−1 (9.40 μM) and subcultured for four times at four different incubation periods (30, 45, 60 and 75 days). By the fourth subculture, irrespective of incubations periods, the explants lost 50% of its shoot formation capacity. Longer incubation resulted on higher rate and total shoots than a shorter incubation, however, the magnitude of that different varied over subcultures. The difference in shoots formation between the explants incubated for 30 and 75 days was 6 shoots at first, increased to 21 at the third and declined to 11 shoots at the fourth subculture. Over a period of 75 days, the majority of shoots formation occurred during the first 30 days (35%) at a rate of 1.5 shoot per week and the last 15 days (40%) at a rate of 3.8 shoot per week. In the period between 30 and 60 days, 15% of shoots at rate of 1.8 and 10% at rate of 1.0 shoot per week formed during the first and the second 15 days interval, respectively. Over four consecutive subcultures, explants incubated for 30 days produced the lowest total (1028 shoots) and that incubated for 75 days produced the highest total (120,917 shoots) from a single explant. No significant differences in total were observed between explants incubated for 45 and 60 days over the first three subcultures. However, by the fourth subculture the total of 60 days (14,288 shoots) was two times higher (7163 shoots) than that of 45 days long incubation.  相似文献   

14.
扎米莲( Zamioculcas zamiifolia) 叶片的植株再生   总被引：4，自引：0，他引：4  
施和平  梁朋 《园艺学报》2003,30(5):621-622
 建立了从扎米莲叶片直接再生植株的有效方法。结果表明, 扎米莲叶片外植体在仅加6-BA 或NAA 或2 ,4-D 的培养基中培养8 周后都不能产生幼芽; 但在含不同浓度6-BA (1.0～2.0 mg/L) 和NAA(0.02～0.2 mg/L) 组合的MS 培养基上培养3 周后开始产生幼芽, 其幼芽分化的最佳培养基为MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L。再生芽在添加IBA 0.5 mg/L 的MS 培养基中生根。  相似文献   

15.
Summary

An 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   

16.
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   

17.
Leaf explants of Caladium ‘Pink Cloud’ were cultured in vitro on MS medium containing various auxins (NAA, IBA, IAA, 2,4,5-T and 2,4-D) in combination with cytokinin (BA). NAA gave the most vigorous in vitro propagation of this plant, and only 15% of the plants were leaf-colour variants on the medium containing 0.5 μmol NAA. Leaf colour variation was observed in all plants regenerated on the medium containing 2,4-D at 0.5–4.5 μmol. In hormone-free medium, only a few leaf-colour variants (6%) occurred, but the rate of plant regeneration was very low. Application of 0.5 μmol NAA together with 4.5 μmol BA seemed to be the most appropriate for in vitro propagation of Caladium ‘Pink Cloud’ with only a few leaf-colour variants.  相似文献   

18.
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   

19.
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  
李媛媛  郭修武  代汉萍  刘海涛 《果树学报》2008,25(6):868-871,F0003
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   

20.
Goldenseal (Hydrastis canadensis L.) is an endangered medicinal plant used to treat sore eyes and mouths, cold and flu and also as a dye. The objective of this study was to develop an efficient in vitro propagation protocol for goldenseal. Significantly more shoots (26 shoots per leaf explants) were induced on a medium containing 2.5 μM thidiazuron (TDZ) and 5.0 μM 1-naphthaleneacetic acid (NAA) than any other treatment. Sub-culturing regenerated shoots on a medium with 5.0 μM 6-benzylaminopurine (BA) induced the maximum rate of shoot multiplication. Growth of the regenerated shoots in a temporary immersion bioreactor resulted in significant increases in biomass, shoot height and shoot multiplication. The regenerated shoots from the temporary immersion bioreactor formed roots when transferred onto a medium with 1.0–2.0 μM indole-3-butyric acid (IBA). Regenerated whole plantlets were acclimatized and maintained in standard greenhouse conditions for further growth. The regeneration protocol developed in this study provides a basis for germplasm conservation and for further investigation of this rare, medicinally important species.  相似文献   

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