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1.
Immature and mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 cultured on MS or N6 nutrient medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), formed embryogenic callus. Induction of embryogenic callus and subsequent somatic embryogenesis was possible at a lower concentration of 2,4-D on N6 than MS medium. Immature embryos were highly totipotent, forming somatic embryos at a higher frequency than mature embryos. Addition of amino acids (L-proline or L-tryptophan) to 2,4-D medium resulted in significant enhancement of embryogenesis on culture of mature embryos. Silver nitrate also supported an increased frequency of embryogenesis. Thus it is possible to have high frequency of somatic embryogenesis on culture of mature embryos, which are available in abundance and with ease than immature embryos. The somatic embryos readily germinated and formed plantlets on hormone-free regeneration medium. The regenerated plantlets were successful on transfer to soil and set seed. 相似文献
2.
Immature and mature zygotic embryos of hexaploid, Triticale var. DT-46 formed an embryogenic callus, with subsequent somatic embryo formation upon subculture to MS (Murashige and Skoog, 1962) or N6 (Chu et al., 1975) nutrient medium supplemented with various concentrations (9.0–22.5 M) of 2,4-dichlorophenoxyacetic acid (2,4-D). Of the two types of explants, embryogenic tissue from immature embryos responded at a higher frequency, to form somatic embryos over the callus surface. Leaf-base segment cultured on to 2,4-D-containing medium formed a tissue which did not form somatic embryos and instead differentiated into shoot-buds. N6 medium proved to be more effective than MS in support of somatic embryogenesis or shoot-bud formation. Regeneration of plantlets occurred on 2,4-D-free basal medium. These in vitro-formed plantlets were successfully transferred to soil and set seed. 相似文献
3.
Summary
Kalopanax pictus (Thunb.) Nakai is a tall tree, and its wood has been used in making furniture, while its stem bark is used for medicinal
purposes. Here, we report on the micropropagation of Kalopanax pictus via somatic embryogenesis. Embryogenic callus was induced from immature zygotic embryos. The frequency embryogenic callus
induction is influenced by days of seed harvest. Callus formation was primarily observed along the radicle tips of zygotic
embryos incubated on Murashige and Skoog (MS) medium with 4.4 μM 2,4-dichlorophenoxyacctic acid (2,4-D). Somatic embryogenesis was observed following transfer of embryogenic callus to MS
medium lacking 2,4-D. Somatic embryos at the cotyledonary stage were obtained after 6 wk following culture. Frequency of conversion
of somatic embryos into plantlets was low (35%) on a hormone-free MS basal medium, but it increased to 61% when the medium
was supplemented with 0.05% charcoal. Gibberellic acid (GA3) treatment markedly enhanced the germination frequency of embryos up to 83%. All plantlets obtained showed 98% survival on
moist peat soil (TKS2) artificial soil matrix. About 30 000 Kalopanax pictus plants were propagated via somatic embryogenesis and grown to 3-yr-old plants. These results indicate that production of
woody medicinal Kalopanax pictus plantlets through somatic embryogenesis can be practically applicable for propagation. 相似文献
4.
Explants of germinating zygotic embryos of Eleutherococcus sessiliflorus,an important medicinal plant, produced somatic embryos directly on Murashige and Skoog (MS) medium with 4.5 M 2,4-D. In addition, embryogenic callus formed at a low frequency (less than 7%) from hypocotyl segments after prolonged culture. High frequency somatic embryogenesis was obtained through cell suspension culture after the cells were transferred to medium lacking 2,4-D. Maturation and germination of embryos was influenced by the sucrose concentration of the medium. At a low concentration of sucrose (1%), maturation and germination of embryos occurred readily. At over 6% sucrose, somatic embryos did not germinate although this could be overcome by GA3 treatment. Cold treatment during acclimatization after transfer to soil enhanced survival. Surviving plantlets produced new sprouts after overwintering in the field. 相似文献
5.
Immature and mature zygotic embryos excised from Feijoa fruits were employed as explants and the effects of NH4
+ and NO3
– ionic concentration in basal LPm culture medium supplemented with 2,4-D (10 M) were evaluated. Moreover, the addition of 4 mM of Asn, Gln, and Arg, and levels of Gln (0 to 8 mM) were tested. The original NH4
+ and NO3
– concentration present in the LPm culture medium supplemented with Gln (4 mM) resulted in the highest somatic embryo number from immature zygotic embryos. For mature zygotic embryos, the addition of Asn, Gln or Arg to the basal LPm culture medium resulted in improved somatic embryogenesis induction. Ten weeks in culture allowed the highest somatic embryo number when mature zygotic embryos were used as explant. Half-strength MS culture medium supplemented with BAP (0.5 M) enhanced the conversion of somatic embryos to plantlets. 相似文献
6.
Summary Pepper (cv. New Mexico — 6 and Rajur Hirapur) plants were regenerated from immature zygotic embryos via direct somatic embryogenesis. Somatic embryos were formed directly, without any intervening callus, on the zygotic embryo apex, embryo axis and cotyledons on Murashige and Skoog's (MS) medium containing 2,4-D (418 M), thidiazuron (10 M) and a high concentration of sucrose (6–10%). The best response was observed on MS medium containing 2,4-D (9 M), coconut water (10%) and high sucrose (8%). The entire process of induction and maturation of the embryos was completed on the same medium. Histological examination indicated that secondary embryogenesis also occurred directly from the primary somatic embryos. Differentiation of embryos was nonsynchronous, and some embryos were swollen and distorted with fasciation. More than 70% of the mature normal somatic embryos germinated readily on MS medium containing GA3 or TDZ, alone and in combination, and following transfer to pots developed into normal plants.Abbreviations CM
Coconut milk
- 2,4-D
2,4-dichlorophonoxyacetic acid
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962) medium
- NAA
napthaleneacetic acid
- TDZ
thidiazuron 相似文献
7.
Kim Suk Weon Cheol Oh Seung In Dong Su Liu Jang Ryol 《Plant Cell, Tissue and Organ Culture》2003,72(3):277-280
Japanese honeysuckle plant (Lonicera japonica Thunb.) is rich in iridoid secologanin and is a potentially useful model for the study of secologanin biosynthesis. Culture conditions for high frequency plant regeneration via somatic embryogenesis from zygotic embryo cultures and zygotic embryo-derived embryogenic cell suspension cultures of this species are described. Mature zygotic embryos formed embryogenic calluses at a frequency of 46.7% when cultured on Murashige and Skoog (MS) medium supplemented with 4.52 M 2,4-dichloro-phenoxyacetic acid (2,4-D). Cell suspension cultures were established with embryogenic calluses using liquid MS medium with 4.52 M 2,4-D. Upon plating onto MS basal medium, embryogenic cell suspension cultures produced numerous somatic embryos, which subsequently developed into plantlets at a frequency of 68%. Regenerated plantlets were transplanted to potting soil and grown to maturity in a greenhouse. 相似文献
8.
Summary Somatic embryogenesis and plant regeneration have been achieved in Nothapodytes foetida, which is known for its rich source of anti-cancer and anti-AIDS alkaloids. Callus cultures were initiated from immature
zygotic embryos cultured on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid, 6-benzyladenine
(BA), and kinetin. MS medium devoid of plant growth regulators favored the development of globular somatic embryos that differentiated
further into plantlets. Plantlet regeneration efficiency was effectively increased on MS medium supplemented with BA. Over
90% of the in vitro plantlets survived when transferred to the soil. Alkaloids were detected in different stages of somatic embryos, regenerated
plantlets, and different parts of the 2-yr-old regenerated plants. The somatic embryos contains camptothecin (0.011% dry weight.
DW) and 9-methoxycamptothecin (0.0028% DW). Two-yearold field-grown plants obtained from somatic embryos were analyzed and
contained higher levels of camptothecin (0.20% DW) and 9-methoxycamptothecin. (0.097% DW) accumulated in roots, followed by
stem and leaves. Alkaloids were quantified and identified by TLC and HPLC. 相似文献
9.
Summary The plant regeneration ability of callus obtained from zygotic embryos of the monocot Alstroemeria spp. was studied. The best explants for somatic embryogenesis were immature zygotic embryos in half-ovules when the endosperm was still soft and white. For 2 genotypes embryogenic callus was induced on callus induction medium with a success rate of 54%. The best callus induction period was 10 weeks. The morphology of embryogenic callus was nodular. Somatic embryos were formed after transfer of the callus to regeneration medium. These somatic embryos revealed later on the typical features of zygotic Alstroemeria embryos. The total duration of the plant regeneration protocol, from inoculation till rooted plantlets ready for transfer to the greenhouse, was 28 weeks.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962)
- NAA
-naphthaleneacetic acid 相似文献
10.
Panax japonicus is one of the important medicinal plants. Here, we established the protocol for plant regeneration of P. japonicus via direct somatic embryogenesis. Somatic embryos were directly obtained from the segments of zygotic embryos on MS medium
with 4.4 μM 2,4-D. Thereafter, somatic embryos were produced by repetitive secondary somatic embryogenesis. The secondary
somatic embryo formation was enhanced by plasmolyzing pretreatment (1.0 M mannitol for 10 h). Frequency of secondary somatic
embryo formation from cotyledon segments was lowered by plasmolyzing pretreatment, but the number of somatic embryos per explants
was greatly increased. Plasmolyzing pretreatment resulted in retardation of embryo growth and required subculture to fresh
medium for further growth of embryos into cotyledonary stage. Without plasmolyzing pretreatment, cotyledonary embryos were
obtained after 8 weeks of culture. All the cotyledonary somatic embryos germinated by 5 μM GA3 treatment, but only 15.3% were germinated on hormone-free medium. After 2 months of culture on 1/2 strength WPM medium, plantlets
produced flowers spontaneously. In the anthers of in vitro flowers, microsporogenesis occurred normally with low number of
pollen grains. 相似文献
11.
Myung Jin Oh Hye Ryun Na Hong-Keun Choi Jang Ryol Liu Suk Weon Kim 《Plant biotechnology reports》2008,2(1):87-92
An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension
cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when
cultured on half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly
with increasing concentrations of 2,4-D up to 3 mg l−1, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryo-derived white friable callus
were established using half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos
and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by
up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting
soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high
frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield,
which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could
be useful for the mass propagation and transformation of selected elite lines. 相似文献
12.
Somatic embryogenesis in semul, Bombax ceiba L. (Bombacaceae) was achieved from immature zygotic embryo explants on MS medium. The cytokinin BA induced somatic embryogenesis at higher frequencies than the auxin 2,4-d. The rate of somatic embryogenesis was inversely related to the concentration of BA. A constant supply of 0.44 M BA was necessary for a high multiplication rate. Conversion of somatic embryos into plantlets is reported. 相似文献
13.
Direct regeneration of somatic embryos was obtained from immature zygotic embryos of Dalbergia latifolia. Immature embryos dissected from green pods 90 d after flowering gave the highest frequency of somatic embryo formation. Preculture on high 2,4-D medium for 4 weeks induced direct somatic embryogenesis, which was expressed during the second culture phase in the presence of low 2,4-D along with a high sucrose concentration. Embryos were separated and transferred to the maturation medium containing MS + 0.5–1.0 mg/L BAP, where embryos developed into plantlets. Somatic embryos failed to convert into complete plants without BAP treatment. This method of direct regeneration of somatic embryos without a callus phase has direct application for genetic manipulation studies.Abbreviations MS
Murashige and Skoog (1962) medium
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- ABA
Abscisic acid
- KIN
Kinetin 相似文献
14.
Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (½MS) supplemented with 2.26, 9.05, and 9.05 μM 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ½MS without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber. 相似文献
15.
Summary Somatic embryos could be induced from embryogenic callus originating from mesocotyl as well as leaf-base segments of Paspalum scrobiculatum on Murashige and Skoog (MS) or Chu et al. (N6) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D; 4.5, 9.0, 18.0, and 22.5 μM). N6 medium was better than MS, for both explants, for high-frequency somatic embryogenesis. Also, mesocotyl tissues were relatively
more totipotent than leaf-base segments. The somatic embryos ‘germinated’ and formed plantlets on transfer of embryogenic
calluses to hormone-free MS or N6 regeneration medium. Embryogenic cultures could be maintained on low hormone medium which readily regenerated to form plantlets
on hormone-free medium. A higher frequency of plantlet formation occurred on MS than on N6 medium. In vitro-formed plantlets were gradually acclimatized in the culture room and on transfer to soil flowered and set seed. Somatic embryogenesis
and plantlet regeneration from mesocotyl and leaf-base segments are potentially simpler systems than regeneration from ‘embryonic’
explants such as immature embryos and unemerged inflorescences. 相似文献
16.
In vitro propagation of loblolly pine via direct somatic organogenesis from mature cotyledons and hypocotyls 总被引:3,自引:0,他引:3
A high-efficiency two-step culture procedure for direct somaticorganogenesis in loblolly pine (Pinus taeda L.) resulting inthe formation of multiple shoot structures induced on cotyledons andhypocotyls of mature zygotic embryos is described. Mature zygoticembryos of eight genotypes of loblolly pine were used as explants toinduce direct somatic organogenesis with this two-step culture method,involving the induction and the differentiation of direct adventitiousshoots. After mature zygotic embryos of eight genotypes of loblolly pinewere cultured on induction medium containing 2,4-dichlorophenoxyaceticacid (2,4-D) or -naphthaleneacetic acid (NAA), 6-benzyladenine(BA), and kinetin for 2–3 weeks, embryos were transferred todifferentiation medium. Adventitious shoot regeneration via directsomatic organogenesis with the frequency of 8.7–27.8% wasobtained from mature zygotic embryo cultures of the genotypes tested.The highest mean number of 32.6 adventitious shoots per mature zygoticembryo was produced from genotype La. The tissue culture protocol of invitro shoot regeneration via direct somatic embryogenesis was optimizedafter examining the periods of the induction culture, chillingtreatment, glutamine concentration, and basic medium levels. Rooting wasachieved on TE medium supplemented with 0.5 mg/l indole-3-butyric acid(IBA), 0.5 mg/l gibberellic acid (GA3), and 1 mg/l6-benzyladenine (BA), and regenerated plantlets were established insoil. These results suggested that adventitious shoot regeneration viadirect somatic organogenesis could be useful for clonal micropropagationof some genotypes of loblolly pine and for establishing a transformationsystem of this coniferous species. 相似文献
17.
Control of direct and indirect somatic embryogenesis by exogenous growth regulators in immature zygotic embryo cultures of rose 总被引:3,自引:0,他引:3
Immature zygotic embryos of rose (Rosa hybrida L.; cv. Sumpath) did not form somatic embryos or embryogenic calluses when cultured on half-strength Murashige and Skoog's medium supplemented with various con-centrations of 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole growth regulator. However, the zygotic embryos produced somatic embryos without an intervening callus phase at a frequency of 27.3% on medium with 4.44 M 6-benzyladenine (BA) alone. Immature zygotic embryos formed embryogenic calluses at a frequency of 25% on medium with a combination of 1.36 M 2,4-D and 4.44 M BA. Upon transfer to medium without growth regulators, embryogenic calluses produced numerous somatic embryos that subsequently developed into plantlets. Somatic embryos were induced directly from immature zygotic embryos, or indirectly via an intervening callus phase, by manipulating the exogenous growth regulators. Plantlets were successfully transplanted to potting soil and grown to maturity in a greenhouse. 相似文献
18.
Mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 on MS or N6 nutrient medium supplemented with various concentrations of 2,4-D (4.5 – 22.5 μM) formed embryogenic callus, which differentiated
into somatic embryos within 5 weeks of culture. The somatic embryos after transfer to hormone-free regeneration medium germinated
and formed plantlets. Of the two nutrient formulations, N6 was relatively better than MS for somatic embryogenesis. A culture for 11 d on 100 μM 2,4-D was essential for the establishment
of an embryogenic callus. Shorter duration, 4-d or 7-d culture on 2,4-D medium, supported some proliferation and subsequent
differentiation into shoot-buds or multiple-shoots, in high-frequency cultures. This is first instance in monocots of a controlled
regeneration response; either somatic embryogenesis or shoot formation.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
19.
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
20.
Somatic embryogenesis and plant regeneration from immature zygotic embryos of<Emphasis Type="Italic"> Cryptomeria japonica</Emphasis> D. Don 总被引:2,自引:0,他引:2
Igasaki T Sato T Akashi N Mohri T Maruyama E Kinoshita I Walter C Shinohara K 《Plant cell reports》2003,22(4):239-243
This report describes the successful plant regeneration via somatic embryogenesis from immature zygotic embryos of Cryptomeria japonica D. Don. For the induction of embryogenic tissue, we determined that the optimal medium contained N6-benzyladenine and 2,4-dichlorophenoxyacetic acid. Immature zygotic embryos that were collected at the end of June yielded embryogenic tissue at the highest frequency. Embryogenic tissues that had proliferated in liquid medium included small and loosely packed cells and elongating or elongated cells. We used ten cell lines to determine the optimal medium for the development of somatic embryos. Induced somatic embryos germinated with synchronous sprouting of cotyledons, hypocotyls and roots. Gibberellin A3 in the germination medium had a positive effect on both the elongation of hypocotyls and the survival of seedlings. The frequencies of induction and germination of somatic embryos differed among the cell lines examined. Most of the seedlings grew normally. This system of somatic embryogenesis required 4–5 months for the regeneration of C. japonica plantlets from immature zygotic embryos.Abbreviations ABA Abscisic acid - BA N6-Benzyladenine - 2,4-D 2,4-Dichlorophenoxyacetic acid - GA3 Gibberellin A3Communicated by F. Sato 相似文献