共查询到20条相似文献,搜索用时 109 毫秒
1.
María Perassolo Carla Verónica Quevedo Ana María Giulietti Julián Rodríguez Talou 《Plant Cell, Tissue and Organ Culture》2011,106(1):153-159
Anthraquinone biosynthesis in Rubia tinctorum L. involves different metabolic routes. Chorismic acid, the end-product of the shikimate pathway, becomes the branch point
between primary and secondary metabolism. It has been proposed that the proline cycle could be coupled with the pentose phosphate
pathway (PPP), since the NADP+ generated by proline reduction from glutamate could act as a cofactor of the first enzymes of the PPP. This pathway generates
erythrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the effect of the
addition of glutamate and two proline analogs, azetidine-2-carboxylic acid and thiazolidine-4-carboxylic acid (T4C), on the
PPP, the proline cycle, and anthraquinone production in R. tinctorum cell suspension cultures. The addition of 5 mM of glutamate enhanced both anthraquinone (up to 30%) and total phenolic content
(12%), which correlated well with proline accumulation. Only the addition of 200 μM of T4C resulted in an increase in anthraquinone
production, which was accompanied by a rise in the proline content. Neither the addition of glutamate nor proline analogs
resulted in the induction of PPP, so this route was not a limiting factor as a carbon donor to the shikimate pathway. 相似文献
2.
Janetsy Borroto Josep Coll Maribel Rivas Maria Blanco Oscar Concepción Yudelsy A. Tandrón Martha Hernández Reinaldo Trujillo 《Plant Cell, Tissue and Organ Culture》2008,94(2):181-187
Morinda royoc L. (Rubiaceae) root cultures were established for the production of anthraquinones (AQs). Three independent experiments were
performed to evaluate the effects of different levels of indole-3-acetic acid (0–22.8 μM), culture duration (15–75 days) and
subculture number (0–4). The following indicators were recorded: root fresh weight per Erlenmeyer and intracellular and extracellular
AQ production. The experiments performed in this study allowed an increase of intracellular AQ content up to a maximum of
4.5 mg g−1 of fresh mass, after 30 days of culture in a medium 5.7 μM of IAA. In addition, isolation and identification of seven AQs
from M. royoc L. roots is described, one of them being reported for the first time for this species. The structures of isolated compounds
were determined from 1H-NMR data. To the best of our knowledge, this is the first report on AQ production from root culture of this plant. 相似文献
3.
The effects of cadmium (Cd) on germination, and antioxidative enzyme activity (AEA) involving superoxide dismutase, catalase,
peroxidase, and ascorbate peroxidase, and on amounts of malondialdehyde and proline present within Achnatherum inebrians, were determined for specimens infected (E+) vs. non-infected (E−) by Neotyphodium gansuense, and cultivated in the presence of various concentrations of CdCl2 (0, 50, 100, 200 and 300 μmol/l). Under high Cd concentrations (100, 200 and 300 μM), E+ (vs. E−) specimens exhibited a higher
germination rate and index, and higher values for shoot length, root length and dry biomass, but there was no significant
difference (P > 0.05) under low Cd concentrations (0 and 50 μM). AEA and the proline content increased, but malondialdehyde content declined
in the E+ (vs. E−) specimens under high Cd concentrations (100, 200 and 300 μM). There was no significant difference (P > 0.05) under low Cd concentrations (0 and 50 μM). Endophyte infection was concluded to be of benefit to the germination
and anti-oxidative mechanisms within A. inebrians under plant exposures to high CdCl2 concentrations. 相似文献
4.
Mary Chebib Navnath Gavande Kit Yee Wong Anna Park Isabella Premoli Kenneth N. Mewett Robin D. Allan Rujee K. Duke Graham A. R. Johnston Jane R. Hanrahan 《Neurochemical research》2009,34(10):1704-1711
GABAC receptors play a role in myopia, memory-related disorders and circadian rhythms signifying a need to develop potent and selective
agents for this class of receptors. Guanidino analogs related to glycine, β-alanine and taurine were evaluated at human ρ1GABAC receptors expressed in Xenopus oocytes using 2-electrode voltage clamp methods. Of the 12 analogs tested, 8 analogs were active as antagonists and the remaining
were inactive. (S)-2-Guanidinopropionic acid (IC50 = 2.2 μM) and guanidinoacetic acid (IC50 = 5.4 μM; K
B = 7.75 μM [pK
B = 5.11 ± 0.06]) were the most potent being competitive antagonists at this receptor. In contrast, the β-alanine and GABA
guanidino analogs showed reduced activity, indicating the distance between the carboxyl carbon and terminal nitrogen of the
guanidino group is critical for activity. Substituting the C2-position of guanidinoacetic acid with various alkyl groups reduced
activity indicating that steric effects may impact on activity. The results of this study contribute to the structure–activity-relationship
profile required in developing novel therapeutic agents. 相似文献
5.
Effect of 24-epibrassinolide on biomass, growth and free proline concentration in Spirulina platensis (Cyanophyta) under NaCl stress 总被引:2,自引:0,他引:2
In this study, biomass, growth and free proline concentration were investigated in Spirulina platensis treated with different concentrations of NaCl (50, 100, 150, and 200 mM) and 24-epibrassinolide (24-epiBL) hormone (0.5,
1.0, and 3.0 μM) over 5 days. As a result of analysing the cultures under salinity stress, it was determined that biomass
and growth rate decreased significantly, while proline concentration increased considerably under salinity stress. The increase
in the concentration of proline suggests a role in response to NaCl stress. Among the cultures treated with different concentrations
of 24-epiBL, maximum growth was determined at the cultures at 1.0 μM 24-epiBL. Algal growth was also greater at the 0.5 and
3.0 μM concentrations of 24-epiBL with respect to control cultures. With respect to control, 24-epiBL affected growth rate
and biomass positively, but proline concentration did not change. Among the cultures supplied with different combinations
of NaCl and 24-epiBL, growth rate increased in 150/0.5 and 150/3.0 mM/μM concentrations, but was maximal for the culture containing
150/1.0 mM/μM combination. The increase in algal growth suggests a role for 24-epiBL in partially alleviated to NaCl stress.
These results suggest that 24-epiBL may have a protective role for S. platensis reducing the inhibitor effects of salinity stress. 相似文献
6.
Tetraploid plants were produced from leaf explants of diploid Populus pseudo-simonii by treating the leaves with colchicine. Leaf explants were cultured on MS basal medium containing 1.78 μM BA and 1.08 μM
NAA for 0, 6 and 12 days, and then transferred to the same MS liquid medium with colchicine at concentrations of 25, 50 and
75 μM for 1, 2 and 3 days. The highest efficiency of tetraploid induction was 14.6% by treating leaf explants that were pre-cultured
for 6 days and then cultured in liquid MS with 50 μM colchicine for 3 days. Flow cytometric analysis was used to screen the
tetraploids out from the regenerated plants and chromosome number counting was employed to confirm the polyploidy level. Size
and frequency of leaf stomata between diploid and tetraploid plants were demonstrated to have significant differences. 相似文献
7.
N. Javadian G. Karimzadeh S. Mahfoozi F. Ghanati 《Russian Journal of Plant Physiology》2010,57(4):540-547
Quantitative changes in total leaf soluble proteins, proline, carbohydrate content, chlorophyll fluorescence, guaiacol peroxidase
(POD) and catalase (CAT) activities were determined in a less cold-hardy (LCH) spring cv. Kohdasht (LT50 = −6°C), a semi cold-hardy (SCH) facultative cv. Azar 2 (LT50 = −15°C), and a cold-hardy (CH) winter cv. Norstar (LT50 = −26°C) of wheat (Triticum aestivum L.) exposed to 4°C for 9 weeks. Seedlings were grown in a controlled growth room for 14 days at 20°C and then transferred
to 4°C (experimental day 0) for 63 days (cold treatment); otherwise they were maintained continuously at 20°C (control treatment).
The samples were harvested 0, 2, 21, 28, 42, and 63 days after exposure to 4°C. The results showed significant low temperature
(LT)-induced accumulation of total soluble proteins, proline, and carbohydrates and elevation in activities of CAT and POD
in leaves of SCH and CH winter cultivars rather than in LCH spring cultivar. In contrast, the chlorophyll fluorescence (F
v/F
m) declined during LT treatment irrespective of cultivar. The results suggest that developmental traits such as vernalization
requirement of wheat affects on cold-tolerance expression system of plants. 相似文献
8.
Effects of the Proline Analog l-Thiazolidine-4-carboxylic Acid on Proline Metabolism 总被引:2,自引:2,他引:0
The effect of various proline analogs on proline oxidation in mitochondria isolated from etiolated barley (Hordeum vulgare) shoots was investigated. Of the analogs tested, only l-thiazolidine-4-carboxylic acid (T4C) was an effective inhibitor. T4C (1 millimolar) inhibited proline (10 millimolar) -dependent 02 uptake an average of 67%. T4C was also oxidized to some degree (12.9 nanoatoms oxygen per minute per milligram protein for 10 millimolar). The effect of T4C on the oxidation of other mitochondrial substrates was also tested. T4C inhibited 1-pyrrolidine-5-carboxylic acid-dependent oxygen uptake slightly (13%), the oxidation of malate plus pyruvate even less (6%), and stimulated the oxidation of succinate (+11%), exogenous NADH (+19%), and citrate (+20%). Thus, inhibition by T4C in mitochondria is relatively specific to proline oxidation. T4C was found to inhibit proline dehydrogenase and not the transport of proline into the matrix. 相似文献
9.
A novel acetyltransferase (Mpr1) found in Saccharomyces cerevisiae (strain 1278b) has been shown to specifically detoxify a proline analog, l-azetidine-2-carboxylic acid (A2C) in yeast cells [M. Shichiri et al. (2001) J Biol Chem 276: 41998–42002]. We investigated whether the yeast MPR1 gene would function similarly in a plant system and if its expression could confer resistance to proline analogs. The MPR1 gene coding sequence driven by two different constitutive promoters, with or without the 5- and 3-noncoding sequence from the MPR1 gene adjacent to the conventional NOS terminator, was transformed into tobacco (Nicotiana tabacum L. cv. Xanthi) plants via Agrobacterium tumefaciens infection. The presence of the yeast 5- and 3-noncoding sequences appeared to increase the likelihood of MPR1 gene expression in the transgenic plants. The kanamycin-selected transgenic plants with a high level of Mpr1 activity grew normally, and their progeny expressed acetyltransferase activity that could utilize A2C, azetidine-3-carboxylic acid and 4-hydroxy-l-proline as substrates. Resistance to A2C, but not to the other two analogs, was exhibited during leaf tissue culture and seed germination. The A2C toxicity to the wild-type plants was reversed by the addition of proline, suggesting that A2C acts as a proline analog. Our studies confirm that MPR1 can function in a similar fashion in tobacco as in yeast to detoxify the toxic proline analog A2C, so it could potentially be used as a new selectable marker for plant transformation. However, our attempts to utilize MPR1 as an efficient selectable marker gene for the A. tumefaciens-mediated transformation of tobacco were unsuccessful.Abbreviations
A2C:
l-Azetidine-2-carboxylic acid
-
A3C:
Azetidine-3-carboxylic acid
-
Hyp:
4-Hydroxy-l-proline
-
hpt:
Hygromycin phosphotransferase II
-
NPTII:
Neomycin phosphotransferase II
Communicated by H. Wang 相似文献
10.
Katarzyna Szafrańska Milena Cvikrová Urszula Kowalska Krystyna Górecka Ryszard Górecki Olga Martincová Krystyna M. Janas 《Acta Physiologiae Plantarum》2011,33(3):851-859
Effect of Cu (0.1, 1, 10, and 100 μM) on the regeneration of carrot (Daucus carota L.) androgenic embryos of var. Feria and 1014 breeding line as well as on polyamines (PAs), proline contents, lipid peroxidation
and Cu accumulation after 16 and 24 weeks was studied. Generally, growth of Feria rosettes was better than that of the 1014
line. Significant increase in Cu content in tissues was observed in both cultures grown at the highest Cu concentration (100 μM).
The dose-dependent increase in proline in the 16-week-old culture of Feria was observed, while in 1014 its level increased
only at the highest applied Cu concentration. On the contrary, in the 24-week-old culture, significant increase in the proline
content were observed at 100 and 10 μM Cu in Feria and in 1014 breeding lines, respectively. The decline in proline content
and decrease in embryogenic ability in the line 1014 grown in the presence of the highest Cu concentration for 24 weeks may
indicate that a certain threshold of intracellular Cu was crossed. Both in Feria and 1014 line, putrescine and spermidine
were the most abundant free PAs. The increased content of proline and higher contents of the constitutive free putrescine
and spermidine in Feria cultivated for 24 weeks at the highest Cu concentration point to better protection of this cultivar.
Thus, it seems that the higher tolerance of Feria to oxidative stress (characterized by lower thiobarbituric acid reactive
substances value) may result from higher constitutive level of PAs. These data confirm the suggestion that variations in PA
levels depend not only on the concentrations of metals tested, but also on plant species and cultivars. The role of PAs and
proline in the carrot cultures treated with Cu is discussed. 相似文献
11.
Rocío Elizabeth Escobar-Guzmán Fernando Hernández-Godínez Octavio Martínez de la Vega Neftalí Ochoa-Alejo 《Plant Cell, Tissue and Organ Culture》2009,96(2):181-189
Eight cultivars and two accessions of Physalis ixocarpa Brot. were tested for their capacity to regenerate embryos and plants from anther cultures. Anthers were pretreated at 4°C
for 2 days and then at 35°C for 8 days in the dark while cultured on MS medium supplemented with 0.045 μM 2,4-D + 0.03 mg l−1 vitamin B12 (MS1) or with 2.26 μM 2,4-D + 0.1 mg l−1 vitamin B12 (MS3). Anther incubation proceeded under a 16 h photoperiod at 25 ± 2°C. Embryo formation occurred after 6 weeks of incubation
in these conditions. Androgenetic responses were cultivar- and culture medium-dependent, with the greatest embryo yields recorded
for cv. Chapingo (36.3%) on MS1 medium, and with wild-type 2 (21.8%) on MS3. Further development of regenerated embryos was
promoted on MS medium supplemented with 0.54 μM NAA, 8.88 μM BA and 50 mg l−1 casein hydrolysate. The regenerated plants were cultured on half-strength mineral salts MS medium with 2.85 μM IAA to enhance
root formation. Rooted plantlets were transferred to pots and acclimatized to the greenhouse. Ploidy analysis of regenerated
plants using flow cytometry revealed 72% diploids, 15% haploids and 7% triploids. AFLP analysis of regenerated plants from
anthers of a single parental plant showed different polymorphic patterns indicating their gametophytic origin. 相似文献
12.
13.
The regeneration of meristematic tissues from sporophytes of Laminaria digitata was studied by protoplast and tissue culture. Sequential treatment of explants in sterile seawater with 1% Betadine for 5 min,
1% commercial bleach for 1–2 min and 2% antibiotic treatment supplemented with 1 μM GeO2 overnight enabled viable explants as high as 55%. Different morphogenetic responses were observed from tissue culture on
media supplemented with plant growth regulators alone or in combination, mainly filamentous calluses up to 50% according to
the media. Dark green compact calluses were observed on two combinations: 4 μM Pi + 2 μM N-(2-chloro-4-pyridyl)-N’-phenylurea
(CPPU) and 0.04 μM Pi + 0.44 μM 6-benzylaminopurine. Thalloid-like structures comparable to adventitious buds were regenerated
on medium supplemented with 4 μM Pi + 0.45 μM zeatin but at low frequency suggesting a strong genotypic effect. Friable calluses
were developed from protoplasts in enriched medium with polyamines and containing 0.40 μM CPPU + 0.45 μM 2,4-dichlorophenoxyacetic
acid. In order to produce protoplasts, a one-step enzymatic protocol was developed and yields reached 22 × 106 protoplasts per gram of fresh weight. 相似文献
14.
Mauji Ram M. A. Khan Prabhakar Jha Salim Khan Usha Kiran M. M. Ahmad Saleem Javed M. Z. Abdin 《Acta Physiologiae Plantarum》2010,32(5):859-866
In vivo modulation of HMG-CoA reductase (HMGR) activity and its impact on artemisinin biosynthesis as well as accumulation
were studied through exogenous supply of labeled HMG-CoA (substrate), labeled MVA (the product), and mevinolin (the competitive
inhibitor) using twigs of Artemisia annua L. plants collected at the pre-flowering stage. By increasing the concentration (2–16 μM) of HMG-CoA (3-14C), incorporation of labeled carbon into artemisinin was enhanced from 7.5 to 17.3 nmol (up to 130%). The incorporation of
label (14C) into MVA and artemisinin was inhibited up to 87.5 and 82.9%, respectively, in the presence of 200 μM mevinolin in incubation
medium containing 12 μM HMG-CoA (3-14C). Interestingly, by increasing the concentration of MVA (2-14C) from 2 to 18 μM, incorporation of label (14C) into artemisinin was enhanced from 10.5 to 35 nmol (up to 233%). When HMG-CoA (3-14C) concentration was increased from 12 to 28 μM in the presence of 150 μM mevinolin, the inhibitions in the incorporation
of label (14C) into MVA and artemisinin were, however, reversed and the labels were found to approach their values in twigs fed with 12 μM
HMG-CoA (3-14C) without mevinolin. In another experiment, 14.2% inhibition in artemisinin accumulation was observed in twigs in the presence
of 175 μM fosmidomycin, the competitive inhibitor of 1-deoxy-d-xylulose 5-phosphate reductase (DXR). HMG-CoA reductase activity and artemisinin accumulation were also increased by 18.6
to 24.5% and 30.7 to 38.4%, respectively, after 12 h of treatment, when growth hormones IAA (100 ppm), GA3 (100 ppm) and IAA + GA3 (50 + 50 ppm) were sprayed on A. annua plants at the pre-flowering stage. The results obtained in this study, hence, demonstrate that the mevalonate pathway is
the major contributor of carbon supply to artemisinin biosynthesis and HMGR limits artemisinin synthesis and its accumulation
in A. annua plants. 相似文献
15.
Chun-Hua Wu Hosakatte Niranjana Murthy Eun-Joo Hahn Kee-Yoeup Paek 《Acta Physiologiae Plantarum》2008,30(6):891-896
We have successfully established the co-culture of ginseng (Panax ginseng C.A. Meyer) and echinacea [Echiancea purpurea (L.) Moench.] adventitious roots for the production secondary metabolites. Adventitious roots of ginseng and echinacea were
cultured in different proportions (5 g L−1; 4:1, 3:2 and 2:1 ginseng and echinacea, respectively) in 5-L capacity airlift bioreactors containing 4 L Murashige and Skoog
medium supplemented with 25 μM indole-3-butyric acid and 50 g sucrose L−1 and maintained at 25°C in the dark for 40 days. Results showed the negative effect of echinacea adventitious roots on the
growth of ginseng roots, however, by limiting the inoculum density of echinacea, it was possible to establish the co-cultures.
To enhance the accumulation of secondary metabolites, co-cultures were treated with 200 μM methyl jasmonate after 30 days
of culture initiation. Methyl jasmonate elicitation promoted the accumulation of ginsenosides in the co-cultures. It was possible
to produce ginsenosides and caffeic acid derivatives in higher amounts by establishing co-cultures with higher inoculum proportion
of ginseng to echinacea (4:1 and 3:2) followed by elicitation treatment. This work demonstrates the effectiveness of interspecies
adventitious root co-cultures for the production of plant secondary metabolites. 相似文献
16.
The effect of tryptophan on the biosynthesis of proline has been investigated. Cells of Daucus carota grown in B5 medium supplemented with 5×10–4M tryptophan acquired the ability to grow in the presence of inhibitory concentrations of azetidine-2-carboxylic acid, an analog of proline. When trp was added to carrot cell cultures at sub-growth inhibiting concentrations, overproduction of intracellular free proline was observed. An increase was also observed for lys, his, ala, leu and phe. Likewise, the addition of asparagine, glutamic acid and phenylalanine to the medium stimulated the intracellular increase of free proline and other amino acids.Abbreviations A2CA
azetidine-2-carboxylic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- 5MT
5-methyltryptophan
- P5C
pyrroline-5-carboxylic acid
- f.wt.
fresh weight
- d.wt.
dry weight 相似文献
17.
18.
We have characterized cytochromes P450, CYP710A13, and CYP710A14, as the sterol C22-desaturase in the moss Physcomitrella patens. GC–MS analyses demonstrated that P. patens accumulated stigmasterol as the major sterol (56–60% of total sterol) and sitosterol to a lesser extent (8–12%); this sterol
profile contrasts with those in higher plants accumulating stigmasterol as a minor component. Recombinant CYP710A13 and CYP710A14
proteins prepared using a baculovirus/insect cell system exhibited the C22-desaturase activity with β-sitosterol to produce
stigmasterol, while campesterol and 24-epi-campesterol were not accepted as the substrates. The K
m values for β-sitosterol of CYP710A13 (1.0 ± 0.043 μM) and CYP710A14 (2.1 ± 0.17 μM) were at comparable levels of those reported
with higher plant CYP710A proteins. In Arabidopsis T87 cells over-expressing CYP710A14, stigmasterol contents reached a level 20- to 72-fold higher than those in the basal
level of T87 cells, confirming the C22-desaturase activity of this P450 enzyme. The occurrence of the end-products together
with the enzymes involved in the last step of the pathway substantiated the presence of an entire sterol biosynthetic pathway
in P. patens, providing evidence for the conservation of the sterol biosynthetic pathway through the evolutionary process of land plants.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
19.
Scott P. Burns Maria Gallo Barry L. Tillman 《In vitro cellular & developmental biology. Plant》2012,48(1):58-66
Agrobacterium-mediated transformation, employing direct shoot organogenesis, allows for mature transgenic plants to be obtained quickly
(3–4 mo). In this study, peanut (Arachis hypogaea L.) cultivars Florida-07, Georgia Green, Georgia Brown, New Mexico Valencia A, and VC-2 were selected to test their shoot
induction response for use in future transformation experiments. Two types of cotyledon explants were examined, those that
previously had an attached embryo axis upon cotyledon separation (explant A) and those that were embryo axis-free upon separation
(explant B). Explants were placed onto a shoot induction medium with N
6-benzyladenine concentrations ranging from 10–80 μM for Florida-07, Georgia Green, and VC-2; 10–20 μM for Georgia Brown; and
10–640 μM for New Mexico Valencia A. Following a 4-wk culture period, explants were visually rated based on a scale of 1–4,
where 1 indicated slight greening, but no growth, and 4 indicated greening, adventitious bud formation, as well as small leaf
expansion. A difference in shoot induction was observed for the cotyledon explants examined (P > t = <0.0001). Explant A had greater shoot induction with a visual rating of 1.8 ± 0.1; explant B had a rating of 1.6 ± 0.1
(P > t = <0.0001). Additionally, cultivars responded to the culture conditions differently (cultivar × N
6-benzyladenine interaction). Georgia Green on 10 μM N
6-benzyladenine produced the most shoot buds (24.6%) and the highest visual rating (2.1), followed by VC-2 on 10 μM N
6-benzyladenine (22.1%, 1.8), New Mexico Valencia A on 640 μM N
6-benzyladenine (21.4%, 1.8), Georgia Brown on 80 μM N
6-benzyladenine (9.0%, 1.7), and Florida-07 on 40 μM N
6-benzyladenine (7.1%, 1.8). Of the tested varieties, Georgia Green, New Mexico Valencia A, and VC-2 were best suited for future
transformation experiments based on their shoot bud production. 相似文献
20.
An efficient micropropagation system via direct shoot organogenesis from hypocotyl segments of Embelia ribes Burm F. was developed. A high frequency (84%) of adventitious shoot induction was obtained on Murashige and Skoog (MS) medium
supplemented with additives (283.85 μM ascorbic acid [AA], 118.96 μM citric acid [CA], 142.33 μM cysteine, and 684.22 μM glutamine)
and 1.13 μM of thidiazuron (TDZ) after 4 weeks following culture. Further development of shoot primordia into well-grown shoots
of 4–5 cm in length was achieved by sub-culturing explants along with shoot primordia on MS medium supplemented with 0.44 μM
benzyl adenine (BA) and 0.49 μM indole butyric acid (IBA) for three sub-culture periods with an interval of 15 days between
them. The highest shoot multiplication was obtained when explants were incubated on MS medium supplemented with 2.2 μM BA
and 0.49 μM IBA in 4 weeks. All in vitro developed shoots, 3–4 cm in length, rooted when grown on half-strength MS basal medium
along with 2.47 μM IBA within 4 weeks. Moreover, 100% of shoots developed roots when these were treated with 4.93 μM IBA for
20 min and then transferred to pots containing soilrite mix and grown in the greenhouse. In vitro and ex vitro rooted plants
showed a survival of 85 and 95% respectively, during hardening in the greenhouse for a 6-week period. 相似文献