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1.
Somatic embryos were initiated from 12 to 15 weeks postanthesis (WPA) zygotic embryos of Cornus florida L. (flowering dogwood) cultured on Murashige-Skoog (MS) or Schenk and Hildebrandt (SH) medium amended with either 3 mg/L 2,4-D or 5 mg/L 2,4-D and 1 mg/L kinetin. White, opaque globular and early cotyledonary stage embryos were formed directly on detached cotyledons from 2 of the 5 trees sampled after 7 weeks of culture. Morphologically mature embryos developed after an additional 4 weeks incubation on medium without growth regulators; however, many of the embryos were fused in pairs along the entire length of the hypocotyl-radicle axis. Indirect embryogenesis was observed from callus cultures initiated from 9 to 15 WPA zygotic embryos. These cultures have continued to produce embryos for 16 months. Many of the embryos formed roots on germination medium, but only 12% formed plantlets and none developed past the first true leaf stage.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- NAA
1-naphthaleneacetic acid
- FPA
Formalin-propionic acid-ethanol (50%)
- WPA
weeks post-anthesis 相似文献
2.
J. J. Rybczyński W. Zduńczyk 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1986,73(2):267-271
Summary A tissue culture of five wild species of the Secale genus, i.e., S. africanum (Stapf.), S. ancestrale (Zhuk.), S. kuprianovii (Grossh), S. segetale (Rosher.), and S. vavilovii (Grossh), from immature embryos of sizes (stages) varying between 1.0 mm to 3.0mm, cultured on MS (1962) mineral nutrient medium supplemented with 0.62 mg/1–5.0 mg/1 of 2,4-D, was established. Initially various types of callus were observed and a correlation between genotype, size of explant and 2,4-D concentration was found. The best embryogenic response was observed when explants were smaller than 1.0 mm. Induction of somatic embryogenesis of 2.0 mm–3.0 mm explants required a higher concentration of 2,4-D. Most embryoids were formed in the presence of 5.0 mg/l of 2,4-D. Secale africanum and S. kuprianovii appeared to have the highest embryogenic capacity among the five investigated species. For embryoids germination to plantlets the MS medium supplemented with GA3 and cytokinins was used. Ultimately, out of the 932 regenerants obtained 364 originated from somatic embryogenesis.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
deGibberellic acid
- BAP
Benzylaminopurine 相似文献
3.
Summary The explant used to initiate embryogenic callus and the growth regulators used in subsequent induction (IM) and embryo development media (EDM) both influenced rate of somatic embryo development and conversion to plantlets in asparagus. Embryogenic callus derived from spear-cross sections (SS), in vitro crowns (IVC) and lateral buds (LB) was cultured on IM of MS salts and vitamins with 2, 4-D or NAA at 0, 0.01, 0.1, 1.0 or 10 mg/l and kinetin at 0, 0.1, 1.0 or 10 mg/l. The auxin 2,4-D at 1–10 mg/l, in combination with kinetin at 0–1 mg/l, in IM induced the highest frequency of embryos after four weeks; callus derived from SS, IVC and LB had means of 394, 382, and 344 small globular embryos, and 4, 11 and 9 bipolar embryos per gram of callus, respectively. After 6 weeks on EDM, 128, 116 and 51 bipolar embryos (4–7 mm in length) occurred per gram callus and 4.5, 1.4 and 2.1 embryos converted for IVC, SS and LB, respectively. NAA at 1–10 mg/l, in combinations with kinetin 0–1 mg/l, yielded means of 64, 175 and 225 small globular embryos per gram callus on IM for SS, IVC and LB, respectively. NAA promoted a higher rate of embryo development: means of 27, 54 and 91 bipolar embryos per gram callus for SS, LB and IVC, respectively, on EDM. There were 0.5, 9.4 and 11.9 plantlets from these respective callus sources. There was no difference between kinetin levels of 0–1 mg/l on callus growth and embryogenesis, whereas, 10 mg/l in IM was inhibitory.Abbreviations 2,4-D
2,4 dichlorophenoxyacetic acid
- EDM
embryo development medium
- IAA
indole-3-acetic acid
- IM
induction media
- IVC
in vitro crowns
- LB
lateral bud
- LS
Linsmaier and Skoog (1965)
- MS
Murashige and Skoog (1962)
- NAA
naphthaleneacetic acid
- SS
spear-cross section 相似文献
4.
水母雪莲体细胞胚胎发生及其植株再生 总被引:7,自引:0,他引:7
水母雪莲(Saussurea medusa Maxim.)茎和叶片的切段接种于MS+2mg/L NAA 0.5mmg/L 6-BA的培养基上,20d后产生黄褐色的愈伤组织,经过几个月的继代培养,愈伤组织仍保持旺盛的增殖能力,但部分由黄褐色逐渐变为红色,将红色愈伤组织转到MS+0.1mg/L NAA+0.2mg/L 6-BA 5mg/L GA3的培养基上,30d后可分化出大量的体细胞胚,体细胞胚成熟后转到1/2MS+0.2mg/L IAA 0.5%活性炭的培养基上,30d后可长出2-4cm的根,带根的小苗经锻炼后移栽到土壤中,成活率达76%,细胞组织学观察表明,发育成熟的体细胞胚具有胚根,胚轴和胚芽的完整结构,具有独立的维管系统。 相似文献
5.
A tissue culture procedure for the regeneration of somatic embryos and plantlets from somatic cells of the soybean Glycine max is described. Bean pods of soybean cv. TGM119 were immersed in liquid nitrogen for 20 minutes. Young embryos were excised from the immature seeds and cultured to form calli. Calli grown from the young embryos were incubated in liquid culture for two weeks. The liquid suspension culture was filtered to obtain single cells. The soybean cells were cultured for one month in a liquid medium in hanging drop cultures for development into proembryoids. The proembryoids were maintained on a solid growth medium for 40 days. The resultant callus tissue was transferred into MS media containing selected combinations and concentrations of 2,4-Dichlorophenoxyacetic acid, Naphthaleneacetic acid, Kinetin, Benzyladenine and Indoleacetic acid. In the presence of Benzyladenine (0.2 mg/l) and Indoleacetic acid (0.01 mg/l), globular and heart shaped somatic embryos were formed on the surface of the calli. Calli containing somatic embryos were transferred into liquid medium and incubated under low light conditions. After six months further incubation, more than 1,000 plantlets and a large number of somatic embryoids at various developmental stages were obtained per flask.Abbreviations KT
kinetin
- CM
coconut milk
- BA
benzyladenine
- NAA
napthalene acetic acid
- IAA
indole acetic acid
- 2,4-D
2,4 dichlorophenoxy acetic acid
- MS
Murashige and Skoog medium 相似文献
6.
A reproducible system for somatic embryogenesis and plantlet formation of
sandalwood has been developed. A high frequency (100%) of somatic embryos
were induced directly from various explants in MS (Murashige and Skoog,
1962) medium with thidiazuron (1 or 2 M) or
indirectly in medium containing 2,4-D plus thidiazuron. Within 8 weeks,
white globular somatic embryos or friable embryogenic tissue developed on
cultured explants. In S. album the globular somatic
embryos were transferred to MS medium supplemented with IAA (6 M) and kinetin (1 and M) where they
developed further, multiplied and maintained friable embryogenic tissue.
After 15-30 d, mature somatic embryos (1-2 mm) with well-developed
cotyledons were separated and subcultured on to medium containing GA3 (6
M) for germination. Once germinated, elongated somatic embryos
(10-20 mm long) grew further in MS supplemented with lower GA3 (3
M). In S. spicatum, the addition of casein
hydrolysate and coconut milk was necessary for plantlet development from
somatic embryos. From histological studies, it appeared that primary
somatic embryos arose from single cells or had a multicellular origin from
the epidermis or cortical parenchyma. Secondary somatic embryos and friable
embryogenic tissue differentiated from groups of proembryogenic cells from
a superficial layer of the primary somatic embryos.Keywords:
Santalum album, Santalum spicatum, somatic
embryogenesis, histological studies.
相似文献
7.
A. Sankhla Tim D. Davis D. Sankhla N. Sankhla A. Upadhyaya S. Joshi 《Plant cell reports》1992,11(7):368-371
After placement on Murashige and Skoog's basal medium supplemented with 3–5 mg/l 2,4-D, immature inflorescence expiants of Echinochloa frumentacea gave rise to three distinct types of callus: a) loosely arranged and soft; b) compact and translucent; c) compact, sticky and mucilaginous. Somatic embryo formation occurred in type b callus in about 18–24 d. Callus types a and c did not produce somatic embryos. The highest percentage of cultures exhibiting somatic embryogenesis occurred on the medium containing 5 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos also formed directly on the inflorescence (without intervening callus formation) in about 15% of the expiants placed on this medium. The addition of paclobutrazol or uniconazole (0.25 or 1 mg/l) to the medium had no influence on the percentage of cultures exhibiting direct somatic embryogenesis, but paclobutrazol slightly increased the mean number of somatic embryos per culture. Many of the callus-derived somatic embryos germinated when subcultured on basal MS medium supplemented with kinetin. Addition of paclobutrazol or uniconazole to the culture medium at 0.25 or 1 mg/l decreased somatic embryo germination and shoot elongation but increased root length and leaf width. Both paclobutrazol and uniconazole increased survival of the plantlets following transplanting to soil. Increased post-transplant survival was accompanied by reduced water loss from plantlets produced on culture media containing triazoles. 相似文献
8.
Direct somatic embryogenesis from in vitro-cultured leaf segments of multiple disease-resistant pepper, Piper colubrinum Link is reported. Somatic embryos were initiated on Murashige and Skoog's (1962) basal medium containing 2.2 μM benzyladenine+0.46
μM kinetin and multiplied profusely through secondary embryogenesis on the same medium. Some 91% somatic embryos converted
into plantlets on MS medium supplemented with 4.4 μM benzyladenine+0.23 μM kinetin and plantlets developed on half-strength
MS+2.4 μM indole-3-butyric acid. Plantlets were hardened, transferred to soil, and 100% of plants survived. Various developmental
stages of somatic embryogenesis were studied using histological methods.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
9.
Anjani Kumar 《Plant Cell, Tissue and Organ Culture》1992,31(1):47-50
Plantlet regeneration through somatic embryogenesis has been achieved in the apocynaceous medicinal treeThevetia peruviana L. Calluses obtained by culturing young leaf discs on MS medium containing 9 M 2,4-dichlorophenoxyacetic acid and 4.6 M kinetin, when subjected to reduced levels of the growth regulators followed by higher cytokinin treatment, produced numerous somatic embryos. Somatic embryos developed into complete plantlets on a medium devoid of growth regulators. An average of 40–50 plantlets were obtained from 50 mg of embryogenic callus. Survival of transplants was 60% under glasshouse conditions. 相似文献
10.
Lilian F. Pateña Luzminda R. Carlos-Refuerzo Ramon C. Barba 《In vitro cellular & developmental biology. Plant》2002,38(2):173-177
Summary The ‘Carabao’ or ‘Manila Super’ mango (Mangifera indica L.), a virtually neglected fruit before the advent of KNO3 flower induction in the early 1970s, is now the third leading Philippine export fruit after banana and pineapple. To apply
biotechnology for improvement, a reliable embryogenesis and regeneration protocol is required. We have developed a protocol
for somatic embryogenesis and plantlet regeneration in mango: eight strains of ‘Carabao’ and two unidentified varieties, PHL
12384 and PHL 12378. Over 40 batches of nucellar explants from immature fruis (0.75–5.0 cm long) were cultured in vitro from April 1999 to April 2000. Two media were used, MMSE. Mango Medium for Somatic Embryo Induction, Proliferation and Germination
and MMPR, Mango Medium for Plantlet Regeneration. These are now routinely used. The protocol is reproducible in 14 other varieties
of mango. Shifting the base medium from Gamborg's B5 medium to our own formulation. BP medium (Barba and Pate?a's formulation)
effectively controlled browning. Browning has limited the successful in vitro culture of many woody species including the mango.
Crop Science Society of the Philippines (CSSP) 2001 Best Paper Award, Asian Agriculture Congress, Westin Philippine Plaza,
Manila, Philippines, April 24–27, 2001 and Philippine Fruit Association 2000 Best Poster Award, 8th National Symposium. PCARRD,
Los Ba?os, Laguna, Philippines, November 14–16, 2000. 相似文献
11.
A. Tirajoh T. S. Kyung Z. K. Punja 《In vitro cellular & developmental biology. Plant》1998,34(3):203-211
Summary Somatic embryogenesis in American ginseng (Panax quinquefolium L.) was investigated from three explant sources (root, leaf and epicotyl) with Murashige and Skoog (MS) medium containing
different growth regulators. Mature roots and leaves obtained from 3- to 5-yr-old field-grown plants, and seedling leaves
and epicotyls from plantlets grownin vitro, were evaluated. From root and epicotyl explants, callus development was optimal with 3,6-dichloro-o-anisic acid (dicamba)
(9.0 μM) and kinetin (KN) (5.0 μM) as the growth regulators. When these calluses were transferred after 3 mo. to dicamba alone (9.0 μM), somatic embryo formation was observed at an average frequency of 15.6% in root explants after an additional 3 mo., and
2% in epicotyl explants after an additional 6 mo. No plantlets were recovered because the embryos germinated to form shoots
with no roots. From leaf explants, callus growth was optimal with α-naphthaleneacetic acid (NAA) at 10.0 μM and 2,4-dichlorophenoxyacetic acid (2,4-D) at 9.0 μM. Somatic embryos developed on this medium, with the highest frequency (40%) obtained after 3 mo. from seedling-leaf explants.
Calluses on mature leaves formed somatic embryos after 7 mo. with NAA/2,4-D at an average frequency of 30%. Transfer of these
somatic embryos to 6-benzyladenine/gibberellic acid (4.4/2.9 μM) promoted shoot development but no roots were observed. Up to 100% of germination was observed within 6 wk on half-strength
MS salts containing activated charcoal (1%) and on NAA/2,4-D (5.0/4.5 μM) with charcoal (1%). On the latter medium, somatic embryos enlarged and frequently gave rise to new somatic embryos after
a brief callusing phase. The embryos germinated through a two-stage process, involving the elongation of the root followed
by the formation of a shoot. The highest recovery of ginseng plantlets from germinated embryos was 61.0%. Following transfer
to potting medium and maintenance under conditions of high humidity and low light intensity, the plantlets elongated and developed
new leaves. A high percentage (50%) of these plants have been acclimatized to soil. 相似文献
12.
Plant regeneration through indirect somatic embryogenesis was attempted from the immature cotyledon-derived explant of Cassia angustifolia Vahl. — a valuable leguminous shrub. The highest frequency (90.5 %) of somatic embryos was obtained on a Murashige and Skoog (MS) medium augmented with 10.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 μM benzyladenine (BA) with the production of a maximum of 22.8 embryos per explant, of which 35.3 % germinated on the same medium after 6 weeks of culture. A half strength MS medium without plant growth regulators facilitated better conversion of embryos into complete plantlets compared to a full strength MS medium. Regenerated plantlets were successfully acclimatized in sterile Soilrite and transferred to field conditions with a 70 % survival rate. Histological studies performed at different stages of embryogenesis revealed the mode of differentiation of embryos from the callus. The content of chlorophylls (a + b) and carotenoids, and the net photosynthetic rate (PN) in the regenerated plantlets were tested during different periods of acclimatization. 相似文献
13.
Somatic embryogenesis of Cyclamen persicum in liquid medium 总被引:1,自引:0,他引:1
Marc Kreuger Erik Postma Yvon Brouwer Gerrit-Jan van Holst 《Physiologia plantarum》1995,94(4):605-612
A method is described for the production of somatic embryos of Cyclamen persicum Mill. in liquid medium. Five steps are involved; initiation of embryogenic cell lines, proliferation of pro-embryogenic masses (PEMs) on auxin-containing medium, development of somatic embryos on hormone-free medium with high osmolarity, germination and subsequent plantlet formation. Cell lines were initiated by culturing the explant, the seedling tuber, directly in liquid medium. Three parameters were important for obtaining embryogenic cell lines; explant density, hormone concentrations and subculture regime. The rate of uptake of the hormones 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin influenced the formation of PEMs. Highly embryogenic cell lines were obtained only when PEMs had formed within 5–7 weeks. PEMs were proliferated for at least 24 months and could be isolated from each subculture for the production of somatic embryos. A high sucrose content (175 m M ) in the development medium without hormones ensured efficient embryo development from PEMs. A subsequent subculture in low sucrose concentration (58 m M ) induced the formation of a tuber, thus promoting germination. Arabinogalactan-proteins (AGPs) from carrot seeds and AGPs bound by the monoclonal antibody ZUM 18 increased the number of PEMs in a culture, showing that the activity of AGPs is not species specific. 相似文献
14.
Summary Embryogenic cell suspension cultures and somatic embryos of five genotypes of beech, were obtained from aged cultures derived from immature zygotic embryos cultured on solid medium containing both 2, 4-dichlorophenoxyacetic acid and N6-benzyladenine. The origin of somatic embryos was traced from single cells. Embryos remained arrested at the globular stage on liquid media, further development was achieved after plating embryogenic aggregates on Murashige and Skoog's medium with half strength major salts supplemented with glutamine and low levels of growth regulators. Cultures of different genotypes showed significant differences in maturation frequency which was not affected by the hormone treatments assayed. The frequency of conversion of embryos into plantlets was low. This frequency increased after cold storage of embryos for up to 7 months.Abbreviations BA
N6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EtOH
ethanol
- GA3
giberrellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog (1962)
- NAA
naphthalene acetic acid
- WPM
woody plant medium (Lloyd and McCown, 1980)
- Z
zeatin 相似文献
15.
Somatic embryos were induced from hypocotyl-derived callus of sesame (Sesamum indicum Var. TMV 6). The influence of different
auxins and cytokinins on somatic embryogenesis was investigated. Among the different auxins tested, 2,4-dichlorophenoxy acetic
acid was the most effective and resulted in the highest frequency of responding cultures and highest average number of somatic
embryo per responding cultures napthaleneacetic acid, indoleacetic acid, indolebutyric acid and 2,4,5-trichlorophenoxypropionic
acid were also effective for embryogenesis, but 2,4,5-trichorophenoxyacetic acid and napthoxyacetic acid were not beneficial.
The combined effect of cytokinins with 2,4-d was also studied. Among the four cytokinins tested, 2.2 chμM benzyladenine with
13.6 chμM 2,4-d slightly enhanced embryogenic efficiency; while kinetin, zeatin, 6-γ-γ-dimethylallylaminopurine enhanced the
frequency of responding cultures. There was a decrease in the number of somatic embryos per culture in the presence of all
cytokinins.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
Heung Kyu Moon Ji Ah Kim So Young Park Yong Wook Kim Ho Duck Kang 《Journal of Plant Biology》2006,49(4):320-325
We tested the possibility of plantlet formation via somatic embryogenesis with leaf segments and mature zygotic embryos from
a rare and endangered tree species,Oplopanax elatus. To induce calli, expiants were cultured under darkness in a solid MS medium containing 3% sucrose, 1g L-1 glutamine, and 0.3% gelrite. Treatment supplements included 2,4-D alone or in combination with thidiazuron. Generally, callus
induction and growth were good from leaf expiants, whereas embryogenic calli could be induced only from zygotic embryos. These
embryogenic calli were white or pale yellow and very friable. ABA and activated charcoal appeared to be important factors
when inducing somatic embryos, with optimum levels being 0.1 mg L-1 and 0.02%, respectively. Many somatic embryos showed abnormalities during their development on the germination medium, but
35% could be converted if placed on a medium containing gibberellic acid (GA3). The germinating embryos sometimes formed secondary embryos at the lower portion of the hypocotyls. Normal or converted
plantlets were acclimatized in an artificial soil mixture; their survival was about 60% after two months. This culturing system
provides a feasible approach for regenerating plants, via somatic embryogenesis, from mature zygotic embryos. 相似文献
17.
Embryogenic callus was initiated from immature zygotic embryos of black pine on medium DCR supplemented with 2 mg 1-1 2,4-D and 0.5 mg 1-1 BAP. The diploid number of chromosomes confirmed the origin of callus from zygotic embryos. The callus was white, glossy, mucilaginous and contained somatic embryos consisted of an embryonic region with densely cytoplasmic cells and suspensor region with long vacuolated cells. Although somatic embryos with green cotyledons were recognisable after ABA treatment and subsequent transfer to growth-regulator free media whole plants have not yet been obtained. 相似文献
18.
In this study, several improvements and simplifications of SE protocols in Pinus pinaster (Ait.), a species of economic importance in the regions of Western Europe, are described. These improvements pertained to all stages of SE including high initiation frequencies in eight control pollinated seed families, relatively high somatic embryo maturation yield when cells were coated with particles of activated charcoal and a rapid production of plants directly in a shade house. The SE initiation frequency from isolated zygotic embryos was high (up to 100%) and plants were produced from 11 embryogenic lines representing all crosses. Based on these results, the estimated number of somatic embryos required to produce 1,000 plants varied from slightly more than the required number of plants to more than double this number depending on the line. Such an estimate is critical in developing plant production strategy when a number of embryogenic lines are considered for production of clonal plants. 相似文献
19.
Initiation of somatic embryogenesis in Pinus banksiana,
P. strobus, P. pinaster, and P. sylvestris at three laboratories
in Canada and France 总被引:1,自引:0,他引:1
Y. S. Park M. A. Lelu-Walter L. Harvengt J. F. Trontin I. MacEacheron K. Klimaszewska J. M. Bonga 《Plant Cell, Tissue and Organ Culture》2006,86(1):87-101
During 2002–2004, three laboratories in Canada and France collaborated to improve initiation of somatic embryogenesis (SE) in jack pine (Pinus banksiana Lamb.), eastern white pine (P. strobus L.), maritime pine (P. pinaster Ait.), and Scots pine (P.␣sylvestris L.), giving particular attention to the effects of (1) N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) versus various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzyladenine (BA), (2) differences in basal nutrient media, i.e., macro- and microelements, and (3) gelling agent concentration. The work was carried out separately at␣each laboratory, but the details of media compositions were shared and tested on their respective species. Results indicate that the developmental stage of the zygotic embryo (ZE) and genotype effects had a large influence on SE initiation, and that genetic effects were consistent over time. Different species responded differently to PGR types and concentration, basal nutrient media, trace elements, and their combinations. Currently, our best initiation rates based on a selected group of genotypes, optimal development stage of ZE, and medium are 3.9% for jack pine, 54.6% for eastern white pine, 76.2% for maritime pine, and 19.7% for Scots pine. 相似文献
20.
P. I. P. Perera D. M. D. Yakandawala V. Hocher J-L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,96(2):171-180
The effect of growth regulators on induction of androgenesis in coconut was investigated using seven different growth regulators
at various concentrations and combinations. Three auxins (1-naphthalene acetic acid—NAA, indoleacetic acid—IAA, picloram)
and three cytokinins (2-isopentyl adenine-2-iP, kinetin, zeatin) were tested either alone or in combination with 2,4-dichlorophenoxyacetic
acid (2,4-D), using modified Eeuwens Y3 liquid medium as the basal medium. Among the tested auxins, 100 μM NAA in combination
with 100 μM 2,4-D enhanced the production of calli/embryos (123) whereas IAA and picloram showed negative and detrimental
effects, respectively, for androgenesis induction over 100 μM 2,4-D alone. Kinetin and 2-iP enhanced the production of calli/embryos
when 100 μM 2,4-D was present in the culture medium. Both cytokinins at 10 μM yielded the highest frequencies of embryos (113
and 93, respectively) whereas zeatin (1 or 2.5 μM) had no impact on microspore embryogenesis. When calli/embryos (produced
from different treatments in different experiments) were sub-cultured in somatic embryo induction medium (Y3 medium containing 66 μM 2,4-D), followed by maturation medium (Y3 medium without growth regulators) and germination medium (Y3 medium containing 5 μM-6-benzyladenine—BA and 0.35 μM gibberellic acid—GA3), plantlets were regenerated at low frequencies (in most treatments ranging from 0% to 7%). 相似文献