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1.
《Scientia Horticulturae》2005,106(4):593-602
A brief culture of mulberry leaves for 8–10 days on MS medium with 18.17 μM TDZ followed by transfer to 8.88 μM BAP supplemented medium triggered high frequency shoot organogenesis (77.6–89.2%) and favoured shoot elongation in Morus spp. Shoot proliferation was highest in the presence of 2.22 μM BAP with induction of 9.4–10.6 shoots per culture. High frequency of root induction (76.0–86.6%) was observed on medium supplemented with 0.49 μM IBA whereas increase in the level of IBA (4.92 μM) resulted in induction of roots along with development of callus from the base of the shoots. The regenerated plants established in soil at higher frequency in rainy season compared to winter and summer. 相似文献
2.
《Scientia Horticulturae》2005,106(2):237-246
In vitro direct multiple shoot formation from seedling explants of Indian high pungent varieties of Capsicum annuum cv. Arka Abhir (AA) and Arka Lohit (AL) was successfully obtained. We were able to induce regeneration potency in these varieties by inverting the explant. Aseptically grown seedling explants with decapitated roots, apical meristem and cotyledonary leaves were inoculated in an inverted position in bud induction medium comprising of Murashige and Skoog's basal medium supplemented with 2-(N-morpholine) ethanesulphonic acid (MES) buffer along with 26.63 μM benzyl adenine (BA), 2.28 μM indole-3-acetic acid (IAA) and 10 μM silver nitrate. Profuse shoot bud induction with 20–25 shoot buds per explant was obtained. Supplementation of phloroglucinol in the bud induction medium resulted in 17 and 18% enhancement in bud induction response in Arka Abhir and Arka Lohit variety, respectively on the inverted hypocotyls. Auxin transport inhibitor tri-iodo benzoic acid (TIBA) in the bud induction medium resulted in induction of buds in a shorter period of 40–45 days when compared to bud induction (BI) medium which takes 55–65 days for bud induction. These buds were transferred to MS medium containing 2.8 μM gibberellic acid and 10 μM silver nitrate resulting in elongation of shoot buds. Transfer of shoots to MS basal medium induced rooting to produce plantlets. This protocol can be efficiently used for mass propagation and presumably also for regeneration of genetically transformed C. annuum tissues. 相似文献
3.
《Scientia Horticulturae》2002,95(3):213-221
Multiple shoots were generated via direct organogenesis on hypocotyl segments excised from in vitro germinated seedlings (45-day-old) of Psidium guajava L. cv. Allahabad Safeda. Modified basal Murashige and Skoog (MMS) medium supplemented with 6-benzylaminopurine (BAP), zeatin or thidiazuron with or without α-naphthalene acetic acid (NAA) were tried. Thidiazuron (0.1 μM) along with 0.54 μM NAA gave the highest response (44.6%) with the regeneration of 3.6 shoots per original explants. These shoots upon subculture gave rise to about 5.0 shootlets per explant in shoot proliferation medium, i.e. MMS supplemented with 2.22 μM BAP + 2.32 μM kinetin. The regenerated microshoots were elongated using a quick dip of gibberellic acid (GA3; 1.44 mM) prior to culture on MMS medium supplemented with 0.88 μM BAP and adenine sulphate (54.29 μM) for 2 weeks. Rooting of microshoots was achieved best on half strength MMS medium supplemented with 4.90 μM indole-3-butyric acid along with 100 mg l−1 activated charcoal. 相似文献
4.
《Scientia Horticulturae》2005,103(3):351-359
Bulb scales of Lilium oriental hybrid ‘Casablanca’ were cultured on MS medium supplemented with cytokinins (kinetin, BA (benzyladenine), TDZ (thidiazuron)). The basal part of bulb scales swelled and formed adventitious shoots with foliage small, leafy bulb scales and abnormally swollen basal plates on the media with cytokinins. Shoots were formed rapidly from the basal parts of bulb scales and became shoot clusters. The medium containing 2.2 μM BA was most favorable in the shoot formation from bulb scales. Cutting shoots into small segments (7–8 mm wide × 12–15 mm length) were cultured on media containing BA and IAA (indole acetic acid) and the segments regenerated many new shoots and formed shoot clusters. The shoot section to shoot cluster cycle method improved adventitious shoot proliferation. The media supplemented with 4.4 μM BA and 5.7 μM IAA. or 8.9 μM BA and 0.6–2.9 μM IAA were effective in allowing the proliferation of shoots from shoot segments under light condition. Sucrose and activated charcoal (AC) improved bulblet growth. Bulblet growth was effectively performed on MS medium containing 60 g/L sucrose. Bulblet growth was also improved by the supplement of AC. The medium with 2.0 g/L AC was most effective for bulblet growth. Normal bulblet growth was stimulated more by the culture of shoots than that of bulb scales. Bulblet weight from shoots reached to an average of over 1100 mg of a bulblet after 3 months in culture on MS medium containing 60 g/L sucrose and 2 g/L AC. Most of the bulblets produced by this method generated stems with several leaves in the green house, after cold treatment at 5 °C for 3 months. 相似文献
5.
《Scientia Horticulturae》2005,104(1):113-120
Single medium based efficient protocol for rapid propagation, and to overcome the delayed rhizome formation in field established in vitro derived plantlets of Kaempferia galanga L. through in vitro rhizome induction was achieved. MS medium with combination of 8.87 μM N6-benzyladenine (BA), and 2.46 μM indole-3-butyric acid (IBA) induced a mean of 6.2 shoots per explant. Addition of 11.7 μM silver nitrate to 8.87 μM BA and 2.46 μM IBA supplemented medium facilitated the highest number of shoots (mean of 8.3 shoots) as well as roots within 60 days. Subculture of isolated shoots on medium with the same concentration of BA, IBA and silver nitrate increased the number to a mean of 12.1 shoots. Silver nitrate enriched medium developed rhizome at the base of shoots. Increase of sucrose concentration (6–8%) in medium with BA, IBA and silver nitrate favoured the best rhizome development. Ninety five per cent of the plantlets survived in field conditions. The plantlets established in field without in vitro developed rhizome (from medium with BA and IBA) did not form rhizome even at 7 months after transplantation. Instead, they developed tuberous roots only. The plantlets with in vitro developed rhizome (on medium having BA, IBA, silver nitrate, and 6–8% sucrose), and that established from conventional way (through splitting of old rhizome) showed no difference in growth of the rhizome. The present study emphasizes the efficacy of silver nitrate and sucrose to develop rhizome in vitro, which enabled to overcome the delayed development of rhizome, and reduced yield of plantlets established in field without in vitro developed rhizome. 相似文献
6.
《Scientia Horticulturae》2001,88(1):41-57
Studies were conducted to improve adventitious bud regeneration in roses (Rosa hybrida L.), specifically to extend the protocol to different genotypes and to initiate production of multiple shoots per explant. The best results were obtained by using a two-stage procedure where excised leaflets were incubated on Murashige and Skoog (MS) (1962) induction medium with 6.8 μM TDZ plus 0.49 μM IBA in the dark for 7 days and subsequently transferred to an MS-based regeneration medium with 2.22 μM BA plus 0.049 μM IBA exposed to a PPFD of 15 μmol m−2 s−1 PAR. Bud formation capacity was also significantly affected by the genotype and the environment, such as the use of bottom cooling creating a lower RH in the vessel. The addition of silver nitrate to the induction medium also significantly improved the percentage of regeneration in three genotypes tested. Regenerated shoots failed to elongate when transferred to MS proliferation medium containing 0.5 mg l−1 BA, however maximum bud development and elongation were achieved when kinetin in the range 1–2 mg l−1 was used. Elongated shoots were excised and rooted best on zero growth regulator half-strength MS modified medium. Rooted plantlets were acclimatized under greenhouse conditions for evaluation of somaclonal variation. 相似文献
7.
《Scientia Horticulturae》2002,95(4):285-295
Multiple shoots were produced from node explants of lime (Citrus aurantifolia (Christm.) Swing) on MS medium supplemented with 6-benzylaminopurine (BAP), 6-furfurylaminopurine (kinetin) and α-naphthaleneacetic acid (NAA). The highest number of shoots, nine shoots per node, were produced on a medium containing 2 mg l−1 BAP (8.8 μM), 1 mg l−1 kinetin (4.6 μM) and 1 mg l−1 NAA (5.4 μM). Depending on the concentration of BAP and kinetin, NAA either inhibited, stimulated or did not affect shoot multiplication, which also depended on the cytokinin level. Maximum shoot length was obtained from treatments containing 0.5 mg l−1 BAP (2.2 μM) combined with 1 mg l−1 kinetin (4.6 μM) and 0.5 mg l−1 NAA (2.7 μM). The largest leaves of resultant shoots were produced on a medium containing 0.5 mg l−1 each of kinetin (2.3 μM) and NAA (2.7 μM). Transferring in vitro shoots to rooting media containing indole-3-butyric acid (IBA) and NAA produced complete plantlets. The highest rooting percentage was obtained on a medium containing either 1 mg l−1 NAA (5.4 μM) alone or 0.5 mg l−1 NAA (2.7 μM) combined with 2 mg l−1 IBA (9.6 μM), whereas the highest number of roots were produced on a treatment containing both 2 mg l−1 NAA (10.8 μM) and 2 mg l−1 IBA (9.6 μM). Roots elongated most on treatments containing 0.5 mg l−1 of either NAA (2.7 μM) or IBA (2.4 μM). Shoot growth associated with the rooting phase was the highest in response to 2 mg l−1 IBA (9.6 μM) or 0.5 mg l−1 NAA (2.7 μM). Plantlets that survived acclimatization, 82%, exhibited normal growth in soil under greenhouse conditions. 相似文献
8.
《Scientia Horticulturae》2002,93(2):143-148
Actively growing shoots of potted greenhouse-grown strawberry tree (Arbutus unedo L.) were initially sterilised and established in basal woody plant medium containing 11.1 μM BA. Optimum shoot proliferation was achieved on a basal WPM containing MS vitamins, sucrose, agar and 22.2 μM BA. Microshoots rooted successfully in basal in vitro medium containing 10 μM IBA or IAA, but their survival rate during acclimatisation was low. Addition of a mixture 1 part peat:4 parts perlite in the basal in vitro rooting medium (1:1 v/v) containing 10 μM IAA resulted in high rooting percentage and plantlets with branched roots. These plantlets were successfully acclimatised. This novel rooting medium can be exploited further due to its potential in commercial applications. 相似文献
9.
A protocol was developed for organogenesis from immature leaflet explants derived from mature seeds of peanut. Immature leaflets pre-incubated on MS medium supplemented with 13.32 μM BAP + 4.95 μM NAA for 7 days, turned green and enlarged. The enlarged green leaflets produced multiple shoot buds after 1–2 cycles of sub-culture on MS medium supplemented with 13.32 μM BAP. Three cycles of shoot buds on the elongation medium (13.32 μM BAP) produced 6.17 ± 0.47 elongated shoots per explant. The shoot bud formation was genotype independent. All elongated shoots rooted on the medium containing 4.95 μM NAA. The complete protocol gave efficient (>81%) direct organogenesis, leading to the development of plantlets within 4 months. 相似文献
10.
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
11.
《Scientia Horticulturae》2005,106(3):415-426
The effect of silver nitrate on shoot differentiation and shoot growth was examined in order to improve the regeneration efficiency of pistachio (Pistacia vera L. cv. Kirmizi) in vitro. Nodal explants of in vitro-grown seedlings were used to test various concentrations and combinations of 6-benzyladenine (BA), kinetin (KIN), gibberellic acid (GA3) and silver nitrate (AgNO3). Addition of AgNO3 up to 48.0 μM to the culture medium improved the regeneration frequency and shoot growth, and reduced basal callus formation in all regenerated explants. The highest regeneration frequency (100%) was recorded on Murashige and Skoog (MS) medium containing 9.0 μM BA, 0.2 μM GA3 and 24.0 or 48.0 μM AgNO3 in combination. The best proliferation response in terms of both shoot formation and low callus production was obtained in the medium containing a combination of 9.0 μM BA, 0.2 μM GA3 and 12.0 μM AgNO3. Regenerated shoots, coming from three cycles of subculturing in proliferation media, were rooted in half-strength MS medium containing 12.0 μM indole-3-butyric acid (IBA). Well rooted plantlets were acclimatized and eventually established in peat and perlite. The development and optimization of an effective micropropagation protocol that is presented in this paper can give an important contribution to improve the quality of pistachio plants and, as a consequence, of orchard production in Middle East countries. 相似文献
12.
《Scientia Horticulturae》2001,87(1-2):131-138
An efficient system has been developed for the in vitro plant regeneration of Lilium longiflorum Thunb. by culturing receptacle sections from flower buds. The sections were cultured on one-half MS medium plus 30 g l−1 sucrose, 8 g l−1 agar, 5.4 μM NAA or 4.9 μM IBA plus 2.2 μM BAP. A section size of 3–4 mm was found to be optimal. After 60 days an average of 41 shoots were formed per explant. More vigorous shoots were obtained by subculturing on hormone-free medium with 20 g l−1 sucrose. Rooting occurred on one-half MS medium with 1.1 μM NAA. Rooted plants were hardened-off in a greenhouse for two months, and normal flowering plants were produced. 相似文献
13.
《Scientia Horticulturae》2005,107(1):64-69
This study describes a protocol for rapid and large-scale in vitro propagation of the valuable medicinal herb Vitis thunbergii Sieb. et Zucc.. Culture conditions influencing shoot proliferation and rooting of the two clones (three- and five-lobed) were examined. Three medium formulations, Murashige-Skoog (MS), Woody Plant Medium (WPM) and Nitsch and Nitsch (NN) medium, were tested for growth of shoot tip culture, and WPM was found to have a superior proliferation rate. The chlorophyll content of leaves was highest in those cultured on NN, followed by those on MS and WPM medium. WPM medium supplemented with 0.5 mg l−1 6-benzyladenine (BA) displayed the highest proliferation rate (15–19 nodes or 3–4 shoots per explant). The rooting was optimized using MS medium supplemented with 0.5 mg l−1 napthaleneacetic (NAA) with eight roots, 3 cm long after 1 month of culture. High frequency callus formation was observed in the basal end of explants cultured on NAA-containing medium. Following acclimatization, rooting plantlets were transferred to the plastic house with a 95% survival rate. 相似文献
14.
《Scientia Horticulturae》2005,106(3):395-401
An efficient and reproducible method for the large-scale propagation of Aerides crispum L. using protocorm and leaf sections has been developed. Protocorm and leaf sections were cultured on Murashige and Skoog (MS) medium supplemented with cytokinins [N6-benzyl adenine (BA), thidiazuron (TDZ), and kinetin (KN), 0.5, 1.0, 2.0 and 5.0 μM], auxins [α-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), 0.5, 1.0, 2.0 and 5.0 μM] and coconut liquid endosperm (CW: 5, 10 and 15%). The explants developed protocorm like bodies (PLBs) within 5–8 weeks on the growth medium. BA supplemented medium was found best for the induction of PLBs and an optimum of 49.1 and 22.0 PLBs developed from protocorm and leaf sections on medium supplemented with 1 and 2 μM BA, respectively. Upon subculture on basal MS medium, the PLBs differentiated plantlets within 6–8 weeks. The resulting plantlets were successfully transferred to potting mixture and 85% of plantlets survived after green house transplantation. This simple protocol will be useful for large-scale propagation of A. crispum L. 相似文献
15.
《Scientia Horticulturae》2001,87(4):319-326
A method is described for producing de novo shoots from leaf derived callus of carnation (Dianthus caryophyllus L.). Plants were regenerated in four steps, viz. callus induction, shoot regeneration, removal of hyperhydricity from regenerated shoots and root development. Callus induction medium contained 2,4-D and BAP. Shoot buds were formed when the callus was further subcultured on 2,4-D- and BAP-containing medium, or MS medium without any growth regulators. The shoots so formed were hyperhydric, bushy in appearance with reduced stem length and watery leaves. The normal conformation of shoots was restored by culturing the hyperhydric shoots onto medium supplemented with GA3 and bactopeptone. The recovered shoots were rooted on MS medium added with NAA (1 mg/l) or IBA (2 mg/l). Regenerated plants with well-developed root and shoot systems were successfully transferred to field conditions after initial acclimation. 相似文献
16.
《Scientia Horticulturae》2005,105(1):117-126
The objectives of the present work were to study the embryogenic competence of floral tissues of Feijoa sellowiana and to investigate the influence of plant growth regulators on somatic embryo induction and development in order to establish a somatic embryogenesis protocol starting from somatic tissues. Petals, stamens and ovaries of floral buds were cultivated onto LPm basal medium supplemented with different levels of 2,4-D, Picloram, 2-iP, Kin and BAP. The highest embryogenic callus induction was obtained with Picloram (10 μM) and Kin (1 μM). Rates of embryogenic calluses induction in stamens and petals were significantly affected by PGRs. Embryogenic calluses were transferred to the same medium, supplemented with gradually reduced levels of PGRs-free medium. After 60 days in suspension cultures with 2,4-D (1 μM) and 2-iP (1 μM) calluses were transferred to PGR-free medium. After 30 days it was observed the development of globular somatic embryos on the surface of 18% of friable calluses previously induced with Picloram (10 μM) and Kin (1 μM). Only embryogenic calluses derived from stamens gave rise to this morphogenetic pattern.Torpedo and cotyledonary somatic embryos transferred to PGR-free culture medium were converted to complete plantlets. This is the first report of somatic embryogenesis in this species starting from somatic tissues. 相似文献
17.
《Scientia Horticulturae》2005,105(3):411-420
Callus induction and somatic embryogenesis of lotus (Nelumbo nucifera Gaertn.) cv. Satabankacha were studied. Callus was initiated by culturing bud, cotyledon, and young leaf explants on Murashige and Skoog (MS) (1962) medium containing a combination of 0, 4, 8 and 10 μM 2,4-dichlorophenoxy acetic acid (2,4-D) and 0, 1, 2 and 3 μM 6-furfuryl amino purine (kinetin) or substituting 0, 0.5 and 1 μM benzyladenine (BA) for kinetin. Bud explants cultured on medium containing 4 μM 2,4-D and 1 μM BA gave the best callus growth. For somatic embryogenesis, the calli initiated on MS medium containing a combination of 4, 6, 8 and 10 μM 2,4-D and 1 μM BA and subsequently transferred to media containing 2–4 μM 2,4-D and 0 or 0.5 μM BA produced the most somatic embryos. When cultures were 12-week-old, callus produced on medium with 6 μM 2,4-D and 1 μM BA showed the best growth for somatic embryo regeneration. When transferred to a medium with 2 μM 2,4-D and 0.5 μM BA somatic embryos were produced from 33% of the calli. Embryos developed to the stage proembryo within 4 weeks and formed globular, heart, torpedo and mature embryos within 16 weeks. 相似文献
18.
《Scientia Horticulturae》2005,107(1):90-96
Somatic embryogenic cell suspension cultures of four ginger cultivars were established. Somatic embryogenic calli were induced from ginger shoot tips on MS agar medium supplemented with 1.0 mg l−1 2,4-D and 0.2 mg l−1 Kn, which contained only half concentration of NH4NO3. Rapid-growing and well-dispersed suspension cultures were established by subculturing this kind of callus in the same liquid MSN medium. The suspension cultures (about 1–2 mm in diameter) were placed on the MSN agar medium for callus proliferation. Thereafter embryogenic callus (1.5 cm2) was transferred to solid media (MS + 0.2 mg l−1 2,4-D + 5.0 mg l−1 BA + 3% sucrose + 0.7% agar). Somatic embryos produced shoots and roots, and shoots developed into complete plantlets on solid MS medium supplemented with 3.0 mg l−1 BA and 0.1 mg l−1 NAA. The relationship between the DW of suspension cultures and pH changes in medium is also discussed. The suspension cultures still kept their vitalities after subculture for 8 months. 相似文献
19.
《Scientia Horticulturae》2005,106(3):427-439
Cell suspension cultures were established from immature cotyledon derived calli from drought tolerant legume horsegram [Macrotyloma uniflorum (Lam.) Verdc.]. Embryogenic callus could be originated from cut slices of the immature cotyledons on MS solid medium [Murashige, T. Skoog, K., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497] augmented with 1.0 μM zeatin and 4.5 μM NAA. Numerous somatic embryoids (26.4%) appeared on MS liquid basal nutrient medium with 5.6 μM NAA and with absence of zeatin during 3 weeks culture. Sustained cell division resulted in the formation of cell aggregates, and then progressed to globular, heart and further if they differentiate properly to torpedo and cotyledonary stages within 5 weeks. Transfer of individual embryos on to a fresh MS basal medium with no plant growth regulators was able to achieve complete maturation. Only a relatively few number of embryos developed into root/shoot when transferred to 0.9 μM GA3, 15 g/l−1 sucrose and 2.4 g/l−1 gelrite containing medium. Substitution of sucrose associated with the use of l-glutamine gave, in the range of concentrations tested, the strongest enhancement of the embryo growth and development. About 5% of somatic embryos were converted into true-to-type fertile plants. 相似文献
20.
《Scientia Horticulturae》2001,89(3):207-215
This study aimed to improve in vitro shoot proliferation efficiency without inducing hyperhydricity in Japanese pear. The shoot number increased at 2.5–10.0 mg l−1 benzyladenine (BA), while shoot fresh mass increased at 1.0 and 2.5 mg l−1 BA. Different macroelement formulation did not affect shoot proliferation, but adding activated charcoal (AC) to the medium inhibited markedly the production of axillary shoots and biomass and many shoots were hyperhydric. Different carbon sources (CS) significantly increased the shoot number and fresh mass, with the best results for shoot proliferation at 20–30 g l−1 sorbitol. With gelling agents, the shoot number increased at 0.4 and 0.6% agar and 0.3% gellan gum, while fresh mass increased at 0.4% agar. The hyperhydric explants were more than 30% at 0.4–0.6% agar and at any concentration of gellan gum. The improved culture (woody plant (WP) supplemented with 20 g l−1 sorbitol, 0.1 mg l−1 3-indolyl-butyric acid (IBA), 2.5 mg l−1 BA and 0.8% agar) and double-phase culture (the same medium using a double-phase liquid-gelling agent solidified culture system) produced a higher number of axillary shoots than the conventional culture (1/2MS supplemented with 0.1 mg l−1 IBA, 1.0 mg l−1 BA, 30 g l−1 sucrose and 0.8% agar), moreover, double-phase culture had a higher fresh mass than the other cultures. 相似文献