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1.
Summary Axillary and terminal buds from suckers of Ananas comosus cv. Phuket were established on Murashige and Tucker-based (MT) medium with 2.0 mgl−1 (9.8 μM) indolebutyric acid, 2.0 mgl−1 (10.74 μM) naphthaleneacetic acid, and 2.0 mgl−1 (9.29 μM) kinetin, followed by multiplication on Murashige and Skoog-based (MS) medium containing 2.0 mgl−1 (8.87 μM) benzyladenine (BA) to provide a continuous supply of axenic shoots. Leaves, excised from such cultured shoots, produced
adventitious shoots from their bases when these explants were cultured on MS medium containing 0.5 mgl−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 mgl−1 (8.87 μM) BA. Embryogenic callus was produced when leaf explants were cultured on MS medium with 3.0 mgl−1 (12.42 μM) 4-amino-3,5,6-trichloropicolinic acid (picloram). Somatic embryos developed into shoots following transfer of embryogenic
tissues to MS medium with 1.0 mgl−1 (4.44 μM) BA. Cell suspensions, initiated by transfer of embryogenic callus to liquid MS medium with 1.0 mgl−1 (4.14 μM) picloram or 1.0 mgl−1 (4.52 μM) 2,4-D, also regenerated shoots by somatic embryogenesis, on transfer of cells to semisolid MS medium with 1.0 mgl−1 (4.44 μM) BA. All regenerated shoots rooted on growth regulator-free MS medium, prior to ex vitro acclimation and transfer to the glasshouse. These studies provide a baseline for propagation, conservation, and genetic manipulation
of elite pineapple germplasms. 相似文献
2.
Wan-Jun Zhang Jiang-Li Dong Ben-Guo Liang Yong-Sheng Jin Tao Wang 《In vitro cellular & developmental biology. Plant》2006,42(2):114-118
Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from
mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl−1 (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl−1 (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl−1 (20.4 μM) 2,4-D and 0.2mgl−1 (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl−1 2,4-D and 0.2mgl−1 Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl−1 Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3%
regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities. 相似文献
3.
Summary High-frequency embryogenesis systems were established for hybrid yellow-poplar (Liriodendron tulipifera×L. chinense) and hybrid sweetgum (Liquidambar styraciflua×L. formosana) by modifying a medium originally developed for embryogenic yellow-poplar cultures. Embryogenic cultures of both hybrids,
consisting of proembryogenic masses (PEMs), were initiated from immature hybrid seeds on an induction-maintenance medium (IMM)
supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), benzyladenine (BA), and casein hydrolyzate (CH). For hybrid yellow-poplar,
as many as 2100 germinable somatic embryos per 4000 cells or cell clumps were produced when PEMs were grown in liquid IMM
lacking CH, at a pH that varied with genotype (3.5 or 5.6), followed by size fractionation and plating on semisolid embryo
development medium (DM; IMM lacking 2,4-D and BA) without CH, but supplemented with 4.0 mgl−1 (15 μM) abscisic acid. For hybrid sweetgum, up to 1650 germinable somatic embryos per 4000 cells or cell clumps were produced when
PEMs were grown in liquid IMM without CH, but with 550 mgl−1
l-glutamine, 510 mg l−1 asparagine, and 170 mg l−1 arginine at pH 5.6. Somatic embryos developed from cell clumps on DM without any plant growth regulators or other supplements.
Hundreds of somatic embryos of both hybrids were germinated on DM without CH, transferred to potting mix, and hardened off
in a humidifying chamber for transfer to the greenhouse. 相似文献
4.
Leaf, stem, hypocotyl, cotyledon, root, shoot tip and embryo explants of Capsicum annuum L. cv. mathania were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) or kinetin (Kin) alone or in
combination with 3-indoleacetic acid (IAA), 3-indolebutyric acid (IBA), α-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic
acid (2,4-D). BAP (5.0 mgl−1) in the medium was found to be the best growth regulator for shoot bud differentiation. Shoot buds cultured on 5.0 mgl−1 BAP increased in number but did not elongate. For obtaining complete plantlets, shoot buds were placed on a medium with IBA
or NAA (0.1 mgl−1). Histological evidence revealed direct differentiation of buds from cotyledons. Regenerated plants were normal diploids.
Unorganized callus could not be induced to differentiate shoot buds. 相似文献
5.
Summary Plant regeneration systems from mesophyll- and cell suspension-derived protoplasts were established in Dianthus acicularis (2n=90), a species with resistance to Burkholderia caryophylli (Psedomonas caryophylli). Protoplasts were isolated from both leaves of in vitro-grown plants and cell suspension cultures established from the calluses originated from leaves of in vitro-grown plants. Protoplasts isolated from both sources showed about the same response to the type and concentration of cytokinins,
and gave the highest frequencies of cell division and colony formation in 0.1% (w/v) Gelrite?-solidified Murashige and Skoog (MS) medium supplemented with 0.5M glucose, 1.0mgl−1 (4.53 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5 mg l−1 (2.28 μM) zeatin. Numerous plantlets were regenerated after transfer of the colonies to 0.8% (w/v) agar-solidified half-strength MS
medium supplemented with 0.5 mgl−1 (2.28 μM) zeatin. Most plantlets exhibited normal phenotypes, but some showed variations, such as abnormal morphology with reduced
chromosome number, precocious flowering, and vigorous growth with a tetraploid chromosome number. Possible mechanisms responsible
for the observed somaclonal variation are discussed. 相似文献
6.
Summary Culture media, environmental and genotypic factors affecting regeneration from multi-shoot cultures derived from corn seedling
apical explants were investigated. The frequency of shoot regeneration was highes for seedlings that were 4–5 cm in length.
Flow cytometry was used to show that the most responsive culturs contained a high proportion of cells in the G1 phase. Proline
in the multi-shoot induction medium (MSI) significantly increased the shoot induction frequency. Continuous low light (30–40
μEm−2s−1) stimulated multi-shoot induction. The highest number of multi-shoots developed in medium containing 4 gl−1 proline, 2 mgl−1 (8.8 μM) 6-benzylaminopurine (BA), and 1 mgl−1 (4.5 μM) 2,4-dichlorophenoxyacetic acid (2,4-D). Multi-shoots were induced in this culture system from 44 of 45 corn genotypes and
approximately 70% of the genotypes exhibited a high to moderate response (greater than 20 shoots per explant in 4 wk of culture).
This culture procedure is an efficient and widely applicable method for corn regeneration that may be a useful target for
transformation. 相似文献
7.
Summary Protoplasts were isolated from Agrobacterium rhizogenes A4-transformed cell line of Medicago sativa L. The highest yield of protoplasts (4.2×106 per g fresh weight) was obtained from 12-d-old calluses after being subeultured on fresh medium. The viability of protoplasts
reached over 80%. Protoplasts were induced to undergo sustained divisions when cultured in Durand et al. (DPD) medium supplemented
with 2 mgl−1 (9.05 μM) 2,4-dichlorophenoxyacetic acid, 0,2mgl−1 (0.93 μM) kinetin, 0.3 M mannitol, 2% (w/v) sucrose, and 500 mgl−1 casein hydrolyzate at a plating density of 1.0×105 per ml. An agarose-beads culture method was appropriate for protoplast division of transformed alfalfa. The division frequency
was about 30%. Numerous hairy roots were induced from protocalluses on Murashige and Skoog medium without growth regulators.
Paper electrophoresis revealed that all of the regenerated hairy roots tested synthesized the corresponding opines. This protoplast
culture system would be valuable for further somatic hybridization in forage legumes. 相似文献
8.
Yuexia Wang Bridget A. Ruemmele Joel M. Chandlee W. Michael Sullivan Jane E. Knapp Albert P. Kausch 《In vitro cellular & developmental biology. Plant》2002,38(5):460-467
Summary Embryogenic callus induction and plant regeneration systems have long been established for creeping bentgrass (Agrostis palustris Huds.), but little research has been reported on optimal medium for embryogenic callus induction and plant regeneration in
velvet bentgrass (Agrostis canina L.), colonial bentgrass (Agrostis capillaries L.), and annual bluegrass (Poa annua L.). The present study compared 14 callus induction media and eight regeneration media for their efficacies on embryogenic
callus induction and plant regeneration in these four species. The embryogenic callus initiation media contained the Murashige
and Skoog inorganic salts and vitamins supplemented with 2,4-dichlorophenoxyacetic acid or 3,6-dichloro-anisic acid and 6-benzyladenine.
l-Proline or casein hydrolyzate was included in some media to stimulate embryogenic callus formation and plant regeneration.
The frequencies of embryogenic callus formation ranged from 0% to 38% and exhibited medium differences within each of the
four species. Callus induction media, plant regeneration media, and genotypes affected plant regeneration rates, which varied
between 0% and 100%. The embryogenic callus induced on Murashige and Skoog medium supplemented with 500 mgl−1 casein hydrolyzate, 6.63 mg l−1 (30 μM) 3,6-dichloro-anisic acid and 0.5–2.0 mg l−1 (2–9 μM) 6-benzyladenine had much higher regeneration rates than those formed on other callus induction media. Embryogenic callus
of annual bluegrass had higher regeneration rates than those of bentgrass species. MSA2D, a media containing 2 mgl−1 (8 μM) 2,4-dichlorophenoxyacetic acid, 100 mgl−1
myo-inositol, and 150 mgl−1 asparagine, was effective in promoting embryogenic callus formation in creeping bentgrass but not in colonial and velvet
bentgrasses and annual bluegrass. 相似文献
9.
Jameel M. Al-Khayri Christine E. Shamblin Edwin J. Anderson 《In vitro cellular & developmental biology. Plant》1996,32(4):227-232
Summary This study was conducted to establish and optimize a regeneration system for adapted U.S. rice genotypes including three commercial
rice cultivars (LaGrue, Katy, and Alan) and two Arkansas breeding lines. Factors evaluated in the study were genotype, sugar
type, and phytohormone concentration. The system consisted of two phases, callus induction and plant regeneration. In the
callus induction phase, mature caryopses were cultured on MS medium containing either 1% sucrose combined with 3% sorbitol
or 4% sucrose alone, and 0.5 to 4 mg·L−1 (2.26 to 18.10 μM) 2,4-D with or without 0.5mg·L−1) (2.32 μM) kinetin. In the plant regeneration phase, callus was transferred to 2,4-D-free MS medium containing 0 or 2 mg·L−1 (9.29 μM) kinetin combined with 0 or 0.1 mg·L−1 (0.54 μM) NAA. Callus induction commenced within a week, independent of the treatments. Callus growth and plant regeneration, however,
were significantly influenced by interactions among experimental factors. Generally, the greatest callus growth and plant
regeneration were obtained with 0.5 mg·L−1 (2.26 μM) 2,4-D and decreased with increasing 2,4-D concentrations. Kinetin enhanced callus growth only when combined with 0.5 mg·L−1 (2.26 μM) 2,4-D, and 4% sucrose. Inducing callus on kinetin-containing medium generally enhanced regeneration capacity in the presence
of sucrose but not with a sucrose/sorbitol combination. Media containing sucrose alone generally supported more callus proliferation,
but the sucrose/sorbitol combination improved regeneration of some cultivars. NAA and kinetin had little effect on regeneration. 相似文献
10.
Calli of P. argentatum were grown on a newly designed liquid nutrient flow-through system which facilitated the subculturing of calli and delayed
browning for 6 weeks. Friable calli were obtained on half-strength Gamborg B5-medium supplemented with 0.05 mgl−1 2,4-dichlorophenoxyacetic acid. Shoots developed on media supplemented with 0.2 mgl−1 benzylaminopurine but lacking 2,4-dichlorophenocyacetic acid. 相似文献
11.
Jianhui Wang Lizhe An Ruoyu Wang Daqun Yang Jing Si Xuanying Fu Jianfeng Chang Shijian Xu 《In vitro cellular & developmental biology. Plant》2006,42(2):148-151
Summary A method was developed for in vitro regeneration of plants via somatic embryogenesis in Chorispora bungeana, an alpine plant with freeze-tolerance, using cell suspensions initiated from leaf-derived callus. Primary calli were induced
from leaves of C. bungeana grown on Murashige and Skoog (MS) media supplemented with 4.0 mg l−1 gibberellic acid (GA3), 0.2 mgl−1 α-naphthaleneacetic acid (NAA) and 0.2 mgl−1 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension culture was initiated by incubating the callus particulates in liquid
MS medium supplemented with 1.0 mgl−1 kinetin (KT) and 0.2 mgl−1 NAA. Individual early cotyledonary-stage somatic embryos isolated from cell suspension developed into whole plants on medium
containing high levels of sucrose (60 and 90 gl−1), whereas lower sucrose concentrations (0 and 30 gl−1) were inhibitory to main root development. On the MS medium with 90 gl−1 sucrose, one regenerated plant exhibited hetero-morphologic leaves, while other plants grown on different media showed a
transformation from stem to root. 相似文献
12.
Anwaar Ahmad Heng Zhong Wengling Wang Mariam B. Sticklen 《In vitro cellular & developmental biology. Plant》2002,38(2):163-167
Summary We report a less genotype-dependent in vitro regeneration system capable of producing multiple shoot clumps and whole plants in four different wheat genotypes. Shool
apical meristems from 7-d-old-seedlings produced axillary and adventitious shoots and somatic embryos on media containing
N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D). All four genotypes responded positively to shoot multiplication
depending upon media composition. Scanning electron microscopies of cultures showed a proliferating budding state that gave
rise to adventitious shoots and somatic embryos on further multiplication. The percentage of relative shoot apical meristem
multiplication was 80–90%, and the average number of shoot meristems per multiplied shoot was 40–50 in all genotypes. Among
different concentrations of phytohormones, 2 and 4 mgl−1 BA (8.8 and 17.7 μM) in combination with 0.5 mg l−1 2,4-D (2.26 μM) gave the best results. Actively multiplying shoot clumps were recovered with high frequency among 3-mo.-old cultures. These
shoot clumps regenerated normally and produced fertile plants containing viable seeds. This in vitro system might prove useful for the production of transgenic plants of wheat in a relatively genotype-independent manner. 相似文献
13.
Summary Few studies have focused on the effect of a broad range of phytohormones on growth and secondary metabolism of a single hairy
root species. We measured growth, development, and production of the antimalarial drug, artemisinin, in Artemisia annua hairy roots in response to the five main hormones: auxins, cytokinins, ethylene, gibberellins (GA), and abscisic acid (ABA).
Single roots grown in six-well plates in medium B5 with 0.01 mgl−1 (0.029 μM) GA3 produced the highest values overall in terms of the number of lateral roots, length of the primary root, lateral root tip
density, total lateral root length, and total root length. When the total root lengths are compared, the best conditions for
stimulating elongation appear to be: GA 0.01 mgl−1 (0.029μM)> ABA 1.0 mgl−1 (3.78μM)=GA 0.02 mgl−1 (0.058μM). Bulk yields of biomass were inversely proportional to the concentration of each hormone tested. All cultures provided with
ABA yielded the highest amount of biomass. Both 6-benzylaminopurine and 2-isopentenyladenine inhibited root growth, however,
only 2-isopentenyladenine stimulated artemisinin production, more than twice that of the B5 controls, and more than any other
hormone studied. These results will prove useful in increasing hairy root growth and artemisinin production. 相似文献
14.
Callus cultures were established from immature embryos of Calotropis gigantea (Linn.) R. Br. on a modified basal medium of Murashige & Skoog supplemented with 1 mgl-1 2,4-D. In addition to 0.1 mgl-1 of NAA the optimal BAP concentration for promoting shoot bud formation and growth was 2 mgl-1. Rooting was induced when shoots were transferred to auxin-supplemented Bonner's solution or half-strength MS basal salt solutions.Abbreviations NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-butyric acid
- BAP
6-benzylaminopurine
- Kin
kinetin 相似文献
15.
Deepak Prem Subhadra Singh Padam Prakash Gupta Jaivir Singh Gaurav Yadav 《In vitro cellular & developmental biology. Plant》2003,39(4):384-387
Summary Guar (Cyamopsis tetragonoloba L. Taub) is a drought-tolerant multipurpose cash crop. A rapid regeneration system has been developed for four Indian guar
genotypes. Investigations were carried out to assess the effect of different growth regulators and their combinations on a
variety of explants such as the embryo, cotyledons, and cotyledonary nodes for shoot morphogenesis. It was established that
Murashige and Skoog's culture medium containing 6-benzylaminopurine (13.3 μM or 3 mgl−1) in combination with indole-3-acetic acid (11.4 μM or 2mgl−1) with cotyledonary node explants gave the highest frequency of multiple shoot induction. In vitro rooting from cultured shoots was maximal on a half-salt concentration of Murashige and Skoog's culture medium fortified with
indole-3-butyric acid (4.9 μM or 1 mgl−1). In vitro-regenerated plants were grown to pod setting and subsequent maturity in greenhouse conditions. 相似文献
16.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
17.
Plant regeneration from protoplast culture of Crocus cancellatus was investigated using regenerable embryogenic calli obtained from shoot meristem culture on LS (Linsmaier and Skoog, 1965)
medium containing 4 mg l−1 kinetin and 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Protoplasts were isolated directly from embryogenic calli. The best protoplast growth
was found on those embedded in Ca-alginate beads and cultured with nurse cells in MS (Murashige and Skoog, 1962) medium supplemented
with 2 mg l−1 kinetin, 1 mg l−1 2,4-D and 100 mg l−1 ascorbic acid at 25 °C in darkness. After 4–5 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads,
but the protoplasts without immobilization in Ca-alginate beads showed very low cell division. Growth of the microcalli in
the medium with nurse cells was much better than in the medium without nurse cells. Transferring beads onto half strength
MS medium supplemented with 0.2 mg l−1 kinetin and 0.1 mg l−1 2,4-D, increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium
growth regulator free or with 1 mg l−1 abscisic acid. Matured embryos germinated on half strength MS medium containing 25 mg l−1 of gibberelic acid. Plantlet formation was obtained on half strength MS medium containing 1 mg l−1 6-benzyladenine and 1 mg l−1 α-naphthaleneacetic acid at 20 °C in a 16/8 h light/dark cycle.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
Summary This is the first communication of direct shoot regeneration from fully developed leaves of potted mature Echinacea purpurea plants. Shoot buds were induced directly on the adaxial surface of mature leaf tissues of E. purpurea 30 d after culture initiation on Woody Plant Medium (WPM) supplemented with various levels of 6-benzyladenine (BA). Maximum
shoot organogenesis, with 12–20 shoots per leaf segment, was obtained with 5% coconut milk and 2.5 mg l−1 (6μM) BA in 30 d. Callus was induced using 0.5 mgl−1 (1μM) α-naphthaleneacetic acid and 2.5 mgl−1 (6μM) BA. The regenerated shoots were rooted on WPM supplemented with 1.5 mgl−1 (3μM) of indole-3-butyric acid, 3% sucrose, and 0.85% agarose. Rooted plants were successfully transferred to soil in pots and
appeared morphologically normal and flowered in a growth chamber. 相似文献
19.
R. V. Sairam C. Wilber J. Franklin B. Smith J. Bazil R. Hassel D. Whaling K. Frutiger C. A. Blakey R. Vierling S. L. Goldman 《In vitro cellular & developmental biology. Plant》2002,38(5):435-440
Summary A protocol for high-frequency callus, somatic embryogenesis, and plant regeneration for Tripsacum is described. Plants were regenerated from complete shoot meristems (3–4 mm) via organogenesis and embryogenesis. In organogenesis,
the shoot meristems were cultured directly on a high cytokinin medium comprising 5–10 mgl−1 (22.2–44.4 μM) 6-benzyladenine (BA). The number of multiple shoots varied from six to eight from each meristem. The time required for production
of plants from organogenesis was rapid (4–6 wk). In contrast, callus was induced on an auxin medium and continuously cultured
on an auxin medium for production of somatic embryos. Prolific callus with numerous somatic embryos developed within 3–4 wk
when cultured on an auxin medium containing 5 mgl−1 (22.6μM), 2,4-dichlorophenoxyacetic acid (2,4-D). The number of shoots induced varied from two to five per callus. Regardless of
the cultivars used, the frequency of callus induction and plant regeneration was between 48% and 94%. The seed germination
procedures also were modified and resulted in a maximum of 60–80% seed germination. Finally, the rate of T-DNA transfer to
complete shoot meristems of Tripsacum was high on the auxin medium and was independent of whether super-virulent strains of Agrobacterium were used or not. 相似文献
20.
Summary Plant regeneration through direct somatic embryogenesis was achieved from root segments derived from in vitro shoots of Rauvolfia micrantha Hook. f. (Apocynaceae) grown for 6 wk in half-strength Murashige and Skoog (MS) medium with 3% sucrose, 100 mgl−1 myo-inositol, and 0.5 mgl−1 α-naphthaleneacetic acid (NAA). The effects of photoperiod and plant growth regulators (PGRs) in half-strength MS medium
were studied for the rapid and maximum induction of somatic embryos. The characteristic globular or heart-shaped stages of
somatic embryogenesis were not found and cotyledonary stage embryos occasionally appeared without the intervention of callus
in total darkness and 16-h photoperiod. Root segments cultured in the medium containing 0.1 mgl−1 NAA and 0.2 mgl−1 6-benzyladenine (BA) under 16-h photoperiod showed the maximum frequency (39%) of embryogenesis. The frequency of embryo
formation was increased to 63% when they were cultured in medium with 0.1 mgl−1 NAA and 0.2 mgl−1 BA in the dark for 4wk, then grown under the 16-h photoperiod. Explants with developing embryos developed into plants after
transfer to half-strength MS medium supplemented with 0.1 mgl−1 BA and 0.05 mgl−1 NAA. The well-developed plants were hardened and most plants (80%) survived and were phenotypically similar to the mother
plants. 相似文献