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1.
Wang Qiaochun Mawassi Munir Sahar Nachman Li Ping Violeta Colova-Tsolova Gafny Ron Sela Ilan Tanne Edna Perl Avihai 《Plant Cell, Tissue and Organ Culture》2004,77(3):267-275
Embryogenic cell suspensions of two grapevine rootstocks: 110 Ritcher (V. berlandieri × V. rupestris), 41B (V. vinifera × V. berlandieri) and several table grape and wine cultivars (Vitis vinifera) were successfully cryopreserved by the encapsulation–vitrification method. Embryogenic cell suspensions were precultured for 3 days in liquid MGN medium supplemented with daily increasing sucrose concentrations of 0.25, 0.5, 0.75 M. Precultured cells were encapsulated and directly dehydrated with a highly concentrated vitrification solution prior to immersion in liquid nitrogen for 1 h. After rewarming at 40 °C for 3 min, cryopreserved cells were post-cultured on solid MGN medium supplemented with 2.5 g l–1 activated charcoal. Surviving cells were transferred to solid MGN medium for regrowth or solid MG medium for embryo development and then to solid WPM for plant regeneration. Optimal viability was 42–76% of cryopreserved cells when cell suspensions were precultured with a final sucrose concentration of 0.75 M and dehydrated with PVS2 at 0 °C for 270 min. Biochemical analysis showed that sucrose preculture caused changes in levels of total soluble protein and sugars in cell suspensions. Although the increase in fresh weight was significantly lower in cryopreserved cells than in control cells, the growth pattern of the cryopreserved cells and control cells was the same after two subcultures, following re-establishment in cell suspensions. Protocol developed in this study suggests a universal and highly efficient cryopreservation system suitable for several genetically diversed Vitis species. 相似文献
2.
Summary An efficient procedure for the in vitro propagation and cryogenic conservation of Syzygium francissi was developed. The maximum number of shoots per explant was obtained on a Murashige and Skoog (MS) medium supplemented with
4.5 μM benzyladenine and 0.5 μM indole-3-butyric acid (IBA). The in vitro-propagated shoots produced roots when transferred to MS medium containing IBA, indold-3-acetic acid, or naphthaleneacetic
acid at various concentrations. Rooted microshoots were transferred to a coco-peat, perlite, and vermiculite (1∶1∶1) mixture,
and hardened off under greenhouse conditions. Ninety-five percent of rooted shoots successfully acclimatized in the greenhouse.
Shoot tips excised from in vitro-grown plants were successfully cryostoraged at −196°C by the encapsulation-dehydration method. A preculture of formed beads
on MS medium containing 0.75 M sucrose for 1 d, followed by 6 h dehydration (20% moisture content) led to the highest survival rate after cryostorage for
1h. This method is a promising technique for in vitro propagation and cryopreservation of shoot tips from in vitro-grown plantlets of S. francissi germplasm. 相似文献
3.
Negash Almaz Krens Frans Schaart Jan Visser Bert 《Plant Cell, Tissue and Organ Culture》2001,66(2):107-111
Studies on in vitro storage of enset under slow-growth conditions were carried out to develop an efficient protocol for conservation of the genetic
diversity of the crop. The response to different growth retardation treatments was examined using three enset clones collected
from southwestern Ethiopia. In vitro cultures could be effectively maintained for 6 months at 15 °C and 18 °C on MS medium supplemented with 10 μM BAP, in the
presence of mannitol at concentrations of 0, 1 or 2% as a growth retardant. Shoots were subsequently recovered and multiplied
on MS medium supplemented with 10 and 20 μM BAP at 25 °C and rooted shoots were successfully transferred to the greenhouse.
Incubation at the lower temperature (15 °C) and the presence of mannitol in the culture medium had a significantly positive
effect on maintenance, measured by the number of recovered shoots after storage.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
T. Dennis Thomas 《In vitro cellular & developmental biology. Plant》2007,43(5):442-448
An efficient and reproducible method for inducing a large number of bulblets from rhizome explants of Curculigo orchioides Gaertn., an endangered medicinal herb, has been developed. The rhizome pieces, measuring about 1 × 1 cm (length × width),
were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of the cytokinins 6-benzylaminopurine,
kinetin, and thidiazuron (TDZ) alone or in combination with 1-naphthalene acetic acid or indole-3-butyric acid (IBA). Of the
three cytokinins used, TDZ at 7 μM gave the maximum response, with 82% of the cultures responding with an average number of
15.4 bulblets per explant. The addition of auxins with cytokinin considerably increased the response. The optimum induction
occurred on MS medium supplemented with 7 μM TDZ and 0.5 μM IBA. On this medium, 88% of the cultures responded with an average
number of 21.4 bulblets per explant. Experiments were also carried out to investigate the role of the sugars sucrose, mannose,
and glucose along with 7 μM TDZ and 0.5 μM IBA. The results indicate that sucrose and mannose at particular concentrations
have critical roles in promoting in vitro bulblet induction. The maximum result was observed on MS medium supplemented with 7 μM TDZ, 0.5 μM IBA, and 200 mM mannose.
On this medium, 97% of the cultures responded with an average number of 26.8 bulblets per culture. Several secondary bulblets
developing from the leaf blades of primary bulblets were produced when the latter were transferred to MS basal medium for
further development. Out of the 45 bulblets transferred to soil, 40 survived. This protocol can be used for the rapid micropropagation
of this endangered medicinal herb. 相似文献
5.
Alexine Braun 《Plant Cell, Tissue and Organ Culture》1988,14(3):161-168
Meristems aseptically isolated from shoots developed on sugarbeet (Beta vulgaris L.) inflorescences were precultured on modified MS agar medium containing 19.4 M 6-benzylaminopurine, 6 M triiodobenzoic acid, and supplemented with 5% DMSO. After two days the meristems were transferred to liquid modified MS medium and the cryoprotectants sorbitol and DMSO added in varying concentrations. The meristems were frozen to –40°C and stored in liquid nitrogen. Growth resumed when the meristems were quick-thawed at 39°C. 相似文献
6.
Z. M. Wei M. Kyo H. Harada 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1986,72(2):252-255
Summary Pollen calli and plantlets of Hordeum vulgare cv. Sabarlis were obtained through direct pollen culture without pretreatment of spikes or preculture of anthers. Isolated immature pollen grains were cultured first in a 0.3 M mannitol solution or a C1 basal medium (Chen et al. 1979) supplemented with 0.3 M mannitol but without sucrose for 5–7 days, then transferred into a C1 medium containing 6% sucrose, 3 mM glutamine and 5 mM m-inositol. After a 3 week culture period small pollen calli derived from the pollen grains were transferred into a growth medium comprising C1 basal medium supplemented with 250 mg/1 lactalbumin hydrolysate and 0.5 mg/1 kinetin. For shoot regeneration, vigorously growing calli were transferred onto agarsolidified MS medium (Murashige and Skoog 1962) containing 3% sucrose, 2 mg/1 benzyladenine and 0.5 mg/1 indole-3-acetic acid. The ratio of green plants to albino was approximately 12.2. 相似文献
7.
Plants were regenerated from root explants of Arabidopsis halleri (L.) O’Kane and Al-Shehbaz via a three-step procedure callus induction, induction of somatic embryos and shoot development. Callus was induced from root segments, leaflets and petiole segments after incubation for 2 weeks in Murashige and Skoog medium (MS) supplemented with 0.5 mg/l−1 (2.26 μM) 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.05 mg/l−1 (0.23 μM) kinetin. Only calli developed from root segments continued to grow when transferred to a regeneration medium containing 2.0 mg/l−1 (9.8 μM) 6-γ-γ-(dimethylallylamino)-purine (2ip) and 0.05 mg/l−1 (2.68 μM) α-naphthalenacetic acid (NAA) and eventually 40 of them developed embryogenic structures. On the same medium 38 of these calli regenerated shoots. Rooting was achieved for 50 of the shoots subcultured in MS medium without hormones. The regeneration ability of callus derived from root cuttings, observed in this study, makes this technique useful for genetic transformation experiments and in vitro culture studies. 相似文献
8.
Summary We describe an in vitro propagation protocol for Zingiber petiolatum (Holttum), I. Theilade, a rare species from the southern part of Thailand. Fruits were surface-sterilized and seeds germinated
on Murashige and Skoog medium (MS) medium supplemented with 3% sucrose. Three-month-old seedlings were used as initial plant
material for in vitro propagation. Terminal buds of the plants were inoculated on MS medium containing 6-benzylaminopurine (BA; 2.2–35.5 μM) alone or in combination with 1-naphthaleneacetic acid (0.5 μM). Eight weeks after inoculation, the cultures were transferred to MS medium without plant growth regulators for 4wk. The
cultures transferred from MS medium with 17.8 μM BA revealed the highest shoot induction rate of 6.1±0.7 shoots per explant. Rooting was spontaneously achieved in MS medium
without plant growth regulators. Rooted plants were successfully transplanted to soil. 相似文献
9.
K. Škrlep M. Bergant G. M. De Winter B. Bohanec J. Žel R. Verpoorte F. Van Iren M. Camloh 《Biologia Plantarum》2008,52(2):329-333
Different lines of cell suspension cultures of Taxus × media Rehd. and Taxus floridana Nutt. were cryopreserved with a two-step freezing method using a simple and inexpensive freezing container instead of a programmable
freezer. Four to seven days old suspension cell cultures were precultured in growth medium supplemented with 0.5 M mannitol
for 2 d. The medium was then replaced with cryoprotectant solution (1 M sucrose, 0.5 M glycerol and 0.5 M dimethylsulfoxide)
and the cells incubated on ice for 1 h. Before being plunged into liquid nitrogen, cells were frozen with a cooling rate of
approximately −1 °C per min to −80 °C. The highest post-thaw cell viability was 90 %. The recovery was line dependent. The
cryopreservation procedure did not alter the nuclear DNA content of the cell lines. The results indicate that cryopreservation
of Taxus cell suspension cultures using inexpensive freezing container is possible. 相似文献
10.
J. B. Kim C. J. J. M. Raemakers E. Jacobsen R. G. F. Visser 《Plant Cell, Tissue and Organ Culture》2006,86(2):233-238
In Alstroemeria high frequencies of compact embryogenic callus (CEC) induction (40%) and friable embryogenic callus (FEC) induction (15%) were obtained from nodes with axil tissue cultured first on a Murashige and Skoog (MS) medium supplemented with 10 μM thidiazuron and 0.5 μM indole-3-butyric acid and after that on a Schenk and Hildebrandt (SH) medium supplemented with 9.1 μM 2,4-dichlorophenoxy acetic acid and 2.2 μM benzylaminopurine (BA). Both types of callus were maintained on modified MS medium supplemented with 20.8 μM picloram. CEC and FEC formed somatic embryos and subsequently plants when transferred to MS medium supplemented with 2.2 μM BA. Plants were produced after 12 weeks (CEC) or after 16 weeks (FEC) of culture. Regenerated plants were established in the greenhouse and flowered normally. 相似文献
11.
A simple, rapid and efficient protocol for micropropagation of Cardiospermum halicacabum via axillary bud multiplication has been successfully developed. The organogenic competence of nodal segments was investigated
on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyladenine (BA), kinetin (Kn), thidiazuron
(TDZ) and 2-isopentenyladenine (2-iP). Multiple shoots differentiated directly without callus mediation within 4 weeks when
explants were cultured on a medium fortified with cytokinins. The maximum number of shoots (14.83 ± 0.52) was developed on
a medium supplemented with 0.3 μM TDZ. Such proliferating shoots when subcultured onto MS media devoid of TDZ gave the highest
rate of shoot multiplication (35.66 ± 1.00) by the end of fourth subculture passage. Elongated shoots were rooted on 1/3 MS
medium augmented with 0.5 μM IAA. The plantlets thus obtained were successfully hardened and transferred to greenhouse. 相似文献
12.
A micropropagation protocol for shoot culture of sweet viburnum (Viburnum odoratissimum) is described. Nodal explants, initially established on MS medium, were transferred to WPM supplemented with combinations of BA and GA3. Maximum shoot multiplication was observed on explants cultured on medium supplemented with BA concentration higher than 1.1 μM, and 14 μM GA3. Although Stage II medium supplemented with BA concentration higher than 1.1 μM resulted in increased shoot multiplication, it also caused a decrease in shoot length. A negative carry over effect of GA3 on rooting was observed in subsequent Stage III cultures. The presence of GA3 in Stage II medium promoted shoot elongation, but it also caused a decrease in microcutting rooting. For this reason, 0.5 μM BA and 14 μM GA3 were selected for optimum Stage II shoot multiplication. Although 100% microcuttings formed roots when cultured on medium containing 6.0 μM NAA, significant callus formation was observed and ex vitro survival rate was low (49%). Rooting was achieved after 3 weeks with 82% of microcuttings on medium supplemented with 3 μM IBA. The survival rate of plantlets under ex vitro conditions was 100% after 3 weeks. Plants looked healthy with no visually detectable phenotypic variation based on observation of about 30 plants. 相似文献
13.
A high-frequency and simple procedure for Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Salvia miltiorrhiza was developed. Leaf discs were pre-cultured on MS medium supplemented with 6.6 μmol l−1 BAP and 0.5 μmol l−1 NAA for one day, then co-cultured with A. tumefaciens strain EHA105 harboring the plasmid pCAMBIA 2301 for three days on the same medium. Regenerated buds were obtained on selection
medium (co-culture medium supplemented with 60 mg l−1 kanamycin and 200 mg l−1 cefotaxime) after two cycles’ culture of 10 days each and then transferred to fresh MS medium with 60 mg l−1 kanamycin for rooting. Fifteen days later, the rooted plantlets were obtained and then successfully transplanted to soil.
The transgenic nature of the regenerated plants was confirmed by PCR, Southern hybridization analysis and GUS histochemical
assay. Averagely, 1.1 independent verified transgenics per explant plated were obtained through this protocol. Adopting this
procedure, positive transformed plants could be obtained within 2–3 months from mature seeds germination to transplant to
soil, and more than 1,000 transgenic plants with several engineered constructs encoding different genes of interest were produced
in our lab in the past two years. 相似文献
14.
Adventitious bud formation in Alhagi graecorum 总被引:1,自引:0,他引:1
Various parts of seedlings and in vitro propagated shoots of Alhagi graecorum Boiss were cultured on different media with different 6-benzyladenine (BA) and kinetin (KIN) concentrations to compare their
potential to regenerate shoots. Murashige and Skoog (MS) medium with 2.5 μM BA and hypocotyl gave the best results. Callus
was obtained from stem segments on MS medium supplemented with 2.5 μM BA, 5 μM 1-naphthaleneacetic acid (NAA) and 0.5 μM 2,4-dichlorophenoxyacetic
acid (2,4-D). Shoot formation from callus occurred upon its transfer to MS medium supplemented with 2.5 μM BA. Mature explants which
showed a relatively low potential for adventitious buds or callus formation, regenerated shoots abundantly using the tiny-mature-explant
method. The regenerated shoots were rooted on half strength MS medium supplemented with 5 μM 3-indolebutyric acid (IBA).
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc
cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings
as well as mature field-grown plants cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (2.27 μM),
6-benzylaminopurine (BA) (2.22 μM) and indole-3-butyric acid (IBA) (0.49 μM). The presence of TDZ in the induction medium
has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction
rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented
with BA (4.44 μM), kinetin (Kn) (2.33 μM), indole-3-acetic acid (IAA) (1.43 μM), and gibberellic acid (GA3) (0.72 μM). Well-developed shoots were rooted on MS medium supplemented with IBA (0.5 μM) after 30 days. Regenerated plants
after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This
protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation. 相似文献
16.
Zhao Yan-Xiu Philip J. C. Harris Yao Dun-Yi 《Plant Cell, Tissue and Organ Culture》1995,40(2):119-123
Protoplasts were isolated from cotyledons of Sesbania bispinosa (Jacq.) W.F. Wight. In a liquid-over-agar culture system with Murashige and Skoog (MS) medium supplemented with 1 mg l-1 2,4-dichlorophenoxyacetic acid (2,4-d, 2 mg l-1 benzyladenine (BA), 1 mg l-1 glutamine and 0.5 and formed callus. The first division occurred after 3–4 days. Callus formed from the protoplasts differentiated shoots by organogenesis on MS medium with 1 mg l-1 indolebutyric acid (IBA) and 1 mg l-1 BA. These shoots developed into complete plantlets when excised and cultured on MS medium with 0.5 mg l-1 IBA. 相似文献
17.
Sompop Prathanturarug Noppamas Soonthornchareonnon Wongsatit Chuakul Yuwalak Phaidee Promjit Saralamp 《In vitro cellular & developmental biology. Plant》2007,43(3):275-279
Node and internode explants of Mallotus repandus were precultured on basal medium (BM: Murashige and Skoog (MS) medium with 3% sucrose and 0.55% Agargel) for 0–18 d before
culture on shoot induction Medium (SIM: BM added with 4.44 μM of benzylaminopurine) for 4 wk. The cultures were subsequently
transferred to BM for 4 wk for shoot elongation. Node explants precultured on BM for 14 d before incubation on SIM were at
an optimum for shoot regeneration with the response rate of 95%, compared to a 21% response for the control without preculture.
Internode explants precultured on BM for 16 d responded with an optimal shoot formation response rate of 69%, whereas the
control response rate was 6%. The maximum shoot regeneration rates were 3.1 ± 0.3 and 2.7 ± 0.4 shoots/responding explant
in node and internode explants, respectively. This study demonstrates for the first time that shoot organogenesis can be induced
from internode explants of M. repandus. Furthermore, the results suggest that the explants need to acquire competence before shoot organogenesis. Rooting was obtained
by incubation of regenerated shoots on half-strength MS with 10.74 μM of 1-naphthylacetic acid for a week before culture on
half-strength MS for 4 wk. Regenerated plants were successfully transferred to soil. 相似文献
18.
The effect of copper sulphate on differentiation and elongation of shoot buds from cotyledonary explants of Capsicum annuum L. cv X-235 was investigated. Shoot buds were induced on medium supplemented with 22.2 μM BAP and 14.7 μM PAA. Elongation
of shoot buds was obtained on MS medium containing 13.3 μM BAP + 0.58 μM GA3. Both shoot induction and elongation media were supplemented with different levels of CuSO4 (0–5 μM). The levels of CuSO4 in the induction as well as elongation medium highly influenced the shoot bud formation and their subsequent elongation.
Highest number of shoot buds per explant was obtained when the concentration of CuSO4 was increased 30 times to the normal MS level. Shoot buds formation frequency i.e., the number of shoots formed per explant
was increased two fold as compared to those formed on control. Elongation both in terms of percentage and length of shoots
was better than that on control. Healthy elongated shoots were rooted on MS medium supplemented with 5.7 μM IAA. Rooted plantlets
were transferred to field conditions. 相似文献
19.
Fast growing calli induced from hypocotyl segments ofGentiana crassicaulis were used for preparation of protoplasts. High yields of viable protoplasts were produced in an enzyme solution containing 1–2% cellulase, I% pecfinase, and 0.5% Hemicellulase. Protoplasts were cultured in KM8P medium containing 1 mg/l 2,4-D, 0.5 mg/l 6BA, 500 mg/l LH, 0.5 M glucose and 0.1 M mannitol by the solid-liquid dual layer culture method. First division occurred within 4–5 days of culture at a frequency of 17.8%. Sustained divisions led to callus formation. Periodically diluting the cultures with freshly prepared liquid medium containing 1% glucose was critical for colony formation. Protocolonies about 2 mm in size were transferred onto MS medium supplemented with 3 mg/l ZT, 2 mg/l 6BA, 1 mg/l GA3, 1 mg/l NAA and 6% sucrose to obtain embryogenic calli. Plantlets were regenerated via somatic embryogenesis at high frequency on hormone-free MS Medium.Abbreviations 6BA
6-benzylaminopurine
- NAA
naphthaleneacetic acid
- 2,4-D
2,4 - dichlorophenoxyacetic acid
- ZT
zeatin
- GA3
gibberellic acid
- LH
lactalbumin hydrolysate
- MES
2-(N-morpholino)-ethane sulfonic acid
- MS
Murashige & Skoog's medium(1962) 相似文献
20.
A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics
of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog
(MS) medium supplemented with 3 μM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated
clearly that the first sign of meristematic cell division could be seen after 1–2 days of pre-culture, and meristematic tissues
multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained
from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 μM
thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for
organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle
bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene. 相似文献