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1.
Experiments were conducted to study plant regeneration through direct somatic embryogenesis using mature zygotic embryo and
cotyledonary explants from seeds of Melia volkensii stored for <3 and >12 months. Explants were cultured on Murashige and Skoog (MS) medium supplemented with BAP, NAA and 2,4-D
(0.5, 1.0 and 2.0 mg l−1) alone, and BAP (0.5, 1.0, 2.0 and 4.0 mg l−1) in combination with 2,4-D or NAA (0.2 and 0.5 mg l−1). After 4 weeks in culture, up to 60% of cotyledonary explants from the seeds stored for <3 months produced direct somatic
embryos on BAP (0.5–4.0 mg l−1) in combination with 2,4-D (0.2 mg l−1). The number of somatic embryos ranged from 5 to 14 per explant in BAP (0.5 mg l−1) and 2,4-D (0.2 mg l−1) combination. Only 20% of cotyledonary explants from seeds stored for >12 months produced somatic embryos. Mature zygotic
embryos failed to produce any somatic embryos. Subcultures of somatic embryos from cotyledonary explants of seeds stored for
<3 months formed clusters of shootlets on semi solid MS and 1/2 MS media. After 6 weeks of subculture on multiplication MS
media augmented with BAP (0.5 mg l−1) and IAA (0.2 mg l−1), 70% of the shoot tips formed 4–7 shoots per explant. Up to 33% of the multiplied shoots were rooted in MS medium supplemented
with 2.0 mg l−1 IBA. Plantlets developed normally into seedlings in the greenhouse. 相似文献
2.
3.
Thidiazuron (TDZ) induced somatic embryogenesis from immature zygotic embryos in Cinnamomum pauciflorum Nees while 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) or picloram only induced callus and/or adventitious
buds. The highest induction frequency for somatic embryogenesis was achieved with MS medium (Murashige and Skoog in Physiol
Plant 15:473–497 1962) supplemented with 2.5 μM TDZ using torpedo-shaped embryos (3–5 mm in length) as explants. In addition, induction medium
was supplemented with 0.8 g l−1 casein, 0.4 g l−1 glutamine, and 10 g l−1 sucrose. Somatic embryos (SEs) initiated from root tips or hypocotyls without callus formation. SEs were maintained and multiplied
via secondary somatic embryogenesis. Embryo maintenance medium was similar to induction medium except that TDZ was reduced
to 0.5 μM. Secondary embryogenesis was enhanced by supplementation of 5 g l−1 activated charcoal in the culture. The best medium for embryo maturation was MS medium containing 30 g l−1 sucrose and 5 g l−1 Phytagel without plant growth regulators. A typical mature SE consisted of two large cotyledons and a short embryo proper.
Approximately 82% of selected mature SEs were able to germinate and 63% could convert into plantlets on germination medium
that was composed of half strength MS medium salts, 10 g l−1 sucrose, 3 g l−1 Phytagel, and 5 g l−1 activated charcoal. 相似文献
4.
Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions. 相似文献
5.
We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue. 相似文献
6.
We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue. 相似文献
7.
We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue. 相似文献
8.
In vitro somatic embryogenesis in callus cultures of Azadirachta indica A. Juss.—a multipurpose tree
Gyana Ranjan Rout 《Journal of Forest Research》2005,10(4):263-267
Plant regeneration via somatic embryogenesis was achieved in embryogenic callus cultures derived from immature zygotic embryos 40 days after anthesis of Azadirachta indica A. Juss. on semisolid basal Murashige and Skoog (MS) salts and vitamins supplemented with 1.11 µM 6-benzylaminopurine (BA) and 4.52–6.78 µM 2,4-dichlorophenoxyacetic acid (2,4-D). The globular-stage embryos were induced when the callus was transferred to medium with 1.11 µM BA and 0.45 µM 2,4-D. The highest average number of somatic embryos per 200 mg of callus was 152.8 after 8 weeks of culture on the medium. Maturation and germination of the somatic embryos were achieved on half-strength MS salts and vitamins supplemented 0.38–0.94 µM abscisic acid (ABA) and 2% (w/v) sucrose. The maximum percentage (64.2%) of germination was obtained with 0.94 µM ABA within 2 weeks of culture. Somatic embryo-derived plantlets were acclimatized in a greenhouse and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones on a mass scale and will also be useful for genetic transformation study. 相似文献
9.
花曲柳体胚发生和植株再生 总被引：1，自引：0，他引：1
以花曲柳合子胚的单片子叶为外植体成功诱导出体胚并获得再生植株。未成熟合子胚的子叶在添加400mg·L-1水解酪蛋白、0.25mg·L-16-BA、1.5mg·L-1NAA、70g.L-1蔗糖和6g·L-1琼脂的MS1/2培养基上可以成功诱导产生体胚,诱导率达到34.7%,每个外植体上体胚数量为2～9个。成熟合子胚的子叶在添加0.25mg·L-16-BA、2mg·L-1NAA的MS1/2培养基(其他成分同上)上可以成功诱导产生体胚,诱导率为10.0%。体胚在MS1/2培养基上经过成熟培养后可以正常萌发,萌发率87.6%。萌发的体胚植株在MS1/2+0.01mg·L-1NAA培养基上生长较好,具备实生幼苗的外观特征。经炼苗后的体胚苗移植到栽培基质(草炭土:蛭石:珍珠岩体积比为5:4:1)中可以正常生长,成活率为75.0%。 相似文献
10.
Somatic embryogenesis in Pinus thunbergii was initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of somatic embryos were obtained on maturation media containing maltose, activated charcoal, abscisic acid, and polyethylene glycol as osmotic agent. The best result among the cell lines tested was achieved with the cell line T-205-3. More than 900 somatic embryos per petri dish, on average, were obtained after about 8 weeks of culture on maturation medium. Sixty percent of somatic embryos tested germinated after transfer to plant growth regulator-free medium and then 85% of them converted into plantlets. 相似文献
11.
This report describes the efficient plant regeneration of Chamaecyparis obtusa Sieb et Zucc. via somatic embryogenesis. Embryogenic cultures were initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated by 2–3-week interval subcultures in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of cotyledonary embryos were obtained on maturation medium containing maltose, polyethylene glycol, activated charcoal, and abscisic acid. Somatic embryos germinated readily after transfer to plant growth regulator-free medium. Growth of regenerated emblings has been monitored in a greenhouse. 相似文献
12.
Somatic embryogenesis was initiated fromCephalotaxus harringtonia (Forbes) K. Koch embryo culture. Explants consisted of embryo and megagametophyte halves both cut longitudinally. They were
removed aseptically from mature seeds and grown together on a solid Murashige and Skoog modified medium supplemented with
5 mg·l
−1 2,4-dichlorophenoxyacetic acid. Embryogenic cultures started from callus after three or more months on the primary medium.
The embryogenic callus originated from the suspensor region of the embryo. All chromosome counts made in the cells of the
embryonic structures demonstrated a diploid stage, which suggest that they originated from zygotic embryo tissue. The early
stages of somatic embryogenic development were achieved,i.e., formation of small clusters consisting of an embryonal region made up of isodiametric meristematic cells. A more advanced
stage was reached in some cultures in which the distal embryonal end of the embryo appeared smooth and opaque. The ultrastructural
characteristics of the embryos, the two types of embryo cells, embryonal and suspensor cells, as well as their contents were
similar to those already reported in the case of somatic embryogenesis of other conifers. 相似文献
13.
Caulogenic calli with a high differentiation potency were induced from mature embryos ofPicea jezoensis seeds stored over a long time, for 29 years, resulting in the active formation of adventitious buds. Embryos began to induce
calli within 3 weeks of cultivating on LP medium containing 3 μM BAP and 1 μM 2,4-D. Then, the calli proliferated and transformed
into caulogenic calli with bud primordia in 8 weeks. The caulogenic calli increased actively with the addition of 500 mg/l ofl-glutamine in the medium. Furthermore, caulogenic calli, induced on LP medium containingl-glutamine, resulted in the formation of adventitious buds, which elongated after transferring the calli into LP medium with
0.1 μM BAP, but withoutl-glutamine. It appears that the number of adventitious buds and the process of shoot elongation are influenced by the kind
of nitrogen contained in the medium for callus induction.
A part of this study was presented at the 107th Annual Meeting of the Japanese Forestry Society (1996). 相似文献
14.
15.
火炬松体细胞胚胎发生和干化体细胞的氧化物酶活性(英文) 总被引：1，自引：0，他引：1
培养于附加2,4-D、BA和KT的愈伤组织诱导培养基上的火炬松成熟合子胚在培养3-9周后形成白色、半透明、有光泽的粘性愈伤组织。这类愈伤组织形成于成熟合子胚的子叶,但当用NAA或者IBA代替愈伤组织诱导培养基中的2,4-D时,它的诱导频率明显降低。这种粘性愈伤组织在分化培养基上形成体细胞胚。体细胞胚经过去50μm ABA和8.5%PEG600处理后成为耐干化胚。扫描电镜观察表明,萌发处理36小时后,耐干化胚恢复到干化处理之前的状态且大小和形态正常,而不耐干化胚不能恢复到干化处理之前的状态且表面撕裂。过氧化物酶活性的分析结果表明,耐干化胚有更高的过氧化物酶活性。耐干化胚的高过氧化物酶活性可能与催化H2O2的分解和保护体细胞胚免受氧化的伤害有关。 相似文献
16.
Rupali Mehta Vikas Sharma Anil Sood Madhu Sharma Ram Kumar Sharma 《European Journal of Forest Research》2011,130(5):729-736
Bambusa nutans Wall., is an evergreen, perennial, and multipurpose bamboo having strong culms, which are largely used for construction,
scaffolding, craft purposes, pulp, and paper industry. Multiple shoots from nodal segments (3–4 cm) of young branches of mature
culms were established in Murashige and Skoog (1962) (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) (1.0–6.0 mg l−1) or in combination with α-naphthaleneacetic acid (NAA) (0.5–1.0 mg l−1) or kinetin (Kn) (1.0–2.0 mg l−1). February–March and December were found to be the best seasons for culture establishments. Maximum shoots were achieved
on MS medium fortified with BAP (2.0 mg l−1). Embryogenic callus (slightly greenish compact, globular, and slow growing) was initiated from the base of severed sprouted
buds in 2–3 subsequent subcultures on MS medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (5.0 mg l−1) under dark incubations. Maturation and germination of well-organized somatic embryos was achieved on MS medium containing
BAP and 2,4-D (1.0 mg l−1 each) with 20.0 mg l−1 ascorbic acid. Full-strength MS medium supplemented with 2% glucose favored further development of proliferated somatic embryos
into plantlets. Genetic variations of field-established B. nutans plants regenerated through tissue cultures were assessed by amplified fragment length polymorphism (AFLP) analysis using
6 primer combinations. Four hundred and seven scorable fragments were amplified, of which 402 (98.8%) have recorded conservation
at various morphogenetic stages leading to plantlets regeneration, therefore, revealed a high level of genetic stability. 相似文献
17.
Embryogenic callus ofQuercus acutissima was successfully induced from embryogenic cultures, and plants were regenerated from the callus. The development of the techniques
involved will allow mass propagation and gene transformation in this species. Embryogenic cultures were formed from embryonic
axis explants (i.e., embryos without cotyledons) excised from immature embryos, after culture on Murashige and Skoog (MS) medium containing indolebutyric
acid and benzyladenine. Attempts to induce embryogenic cultures from cotyledon explants were unsuccessful. Embryogenic calli
were induced at high frequency from embryogenic cultures on MS medium containing 2,4-dichlorophenoxyacetic acid. However,
benzyladenine inhibited embryogenic callus formation. Somatic embryo development from embryogenic calli occurred on MS medium
in all of the seven cell lines tested. Germination of somatic embryos was induced on half strength MS medium without plant
growth regulators. Finally, acclimated plants growing in soil were obtained. 相似文献
18.
唐巍 《中国林学(英文版)》2001,3(2)
三个基固型的火炬松成熟合子胚被培养在附加 8mg·L-12 ,4 D ,4mg·L-1BA ,4mg·L-1KT ,5 0 0mg·L-1水解酪蛋白和 5 0 0mg·L-1谷氨酰胺的愈伤组织诱导培养基上诱导愈伤组织 .来自于子叶、胚轴和胚根的愈伤组织在附加 1 6mg·L-12 ,4 D ,0 8mg·L-1BA和 0 8mg·L-1KT的愈伤组织增殖培养基上培养 9周后 ,可获得 16 9%的胚性愈伤组织 .通过建立胚性细胞悬浮系和研究ABA、PEG和活性炭对体细胞胚成熟的促进作用 ,优化的体细胞胚胎发生体系被建立 .71棵再生小苗被用于移栽试验 ,2 3棵小苗在田间移栽成活 相似文献
19.
Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium
ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the
cell density of 9×104/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of
2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated
from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and
plantlets were regenerated on 1/2 MS medium without a hormone.
A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995). 相似文献
20.
Somatic embryogenesis (SE) in Eucalyptus spp. has been limited to germinated seeds, flowers, lignotubers or zygotic embryos. The low yield of somatic embryos from leaf explants has hampered progress, even though leaves offer a more viable source of clonal explants from superior selected genotypes. It was hypothesised that SE from leaf explants could be enhanced through pairing of synergistic exogenous plant growth regulators, such as natural auxins with natural cytokinins. Callus and embryo induction using 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthalene acetic acid (NAA), indole acetic acid (IAA), and indole butyric acid (IBA), each at either 1.0 or 3.0?mg L?1, indicated that IAA and IBA favoured significantly higher numbers of embryos compared with 2,4-D or NAA. Hence, IAA and IBA were used for subsequent experiments, combining them (at 1.0?mg L?1) with either the synthetic cytokinin, kinetin, or the natural cytokinin, trans-zeatin, both at 0.1?mg L?1. The combination of trans-zeatin and either IAA or IBA resulted in a significant increase in SE (e.g. 86 ± 17.2% and 23 ± 3.2% for IAA with trans-zeatin and kinetin, respectively), compared with kinetin, or with these auxins alone. Embryo maturation and plantlet regeneration was highest in those calli that were induced with IAA and trans-zeatin, indicating that maturation was dependent on auxin depletion, based on the stability of the analogue used for induction. For the E. grandis clone under study, the use of synergistic plant growth regulators significantly enhanced SE from leaf explants, thus presenting the opportunity to benefit from the advantages that SE offers over conventional organogenesis. 相似文献