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1.
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and petiole expiants has been developed in sweetpotato [Ipomoea batatas L. (Lam.)]. The optimal somatic embryogenic response was obtained in the genotype PI 318846-3 with a two-step protocol: (1) stage I-incubation of expiants in the dark for 2 weeks on Murashige Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l) and 6-benzylaminopurine (0.25 mg/l) and, (2) stage II-culture in the light on MS medium with abscisic acid (ABA) (2.5 mg/l). The addition of ABA was critical for enhanced production of somatic embryos. Secondary somatic embryos were produced from the primary embryos cultured on MS medium with 2,4-D at 0.2 mg/l. The somatic embryos were converted into normal plantlets when cultured on basal MS medium. Upon transfer to soil, plants grew well and appeared normal with no mortality. The system of somatic embryogenesis described here will facilitate tissue culture, germplasm conservation and gene transfer research of sweetpotato due to its rapidity (6 to 10 weeks), prolific plant production by direct embryogenesis, ease of secondary somatic embryo production and reproducibility.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine, 2,4-D-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- KIN
kinetin
- MS
medium of Murashige and Skoog (1962)
- NAA
1-naph-thaleneacetic acid
- PIC
picolinic acid
- TDZ
thidiazuron 相似文献
2.
We used four cultivars ofCapsicum annuum L.—Sweet Banana, California Wonder, Yolo Wonder, and Ace—to reexamine the critical factors influencing somatic embryogenesis
from zygotic embryo explants, as reported in the literature. When we followed the protocol of Buyukalaca and Mavituna (1996),
which had induced somatic embryogenesis from mature zygotic embryos of cv. Ace, only callus was formed without embryogenesis
from our mature zygotic embryo expiants. Using the procedures of Harini and Lakshmi Sita (1993) and Binzel et al. (1996),
with some modifications, we were able to induce somatic embryogenesis in all four cultivars. Rates of conversion were significantly
reduced, from 75% and 65% to 40% and 28% in ’Sweet Banana’ and ’California Wonder’, respectively, when the immature zygotic
embryo expiants were held on the induction medium for longer than two weeks. Likewise, somatic embryogenesis of ’Yolo Wonder’
was not observed if the induction medium was supplemented with 10% glucose or fructose, or without 10% sucrose. For somatic
embryo induction and eventual plantlet conversion in Yolo Wonder’, maltose could adequately replace sucrose. In all four cultivars,
somatic embryos were initiated from immature zygotic explants on media with or without coconut water, under both light and
dark conditions. 相似文献
3.
Somatic embryogenesis from pea embryos and shoot apices 总被引:3,自引:0,他引:3
Conditions were defined for plant regeneration via somatic embryogenesis in pea, using explants from immature zygotic embryos or from shoot apices. For the induction of somatic embryos, an auxin (picloram or 2,4-dichlorophenoxyacetic acid) was required. Embryogenic callus originated from embryonic axis tissue of immature embryos and from the axillary-bud region and the plumula of shoot apices. A clear effect of embryo size on somatic embryogenesis was shown. There were differences in frequency of somatic embryogenesis among the five genotypes used in the study. Additions of BA to auxin-containing medium reduced embryo production. Histological examinations confirmed the embryogenic nature of the immature embryo cultures and revealed that somatic embryos originated from the meristematic areas near the callus surface.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
4.
In vitro somatic embryogenesis of Glehnia littoralis Fr. schm. was observed when zygotic embryos were cultured on a medium
containing 1-naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid (0.01-10 μM), with 1 μM being the optimum. Microscopic
observations revealed globular, heart-shaped and torpedo-shaped embryo formations and plantlet regeneration. These somatic
embryos seemed to be produced directly from cells of the zygotic embryos used as explants. Of seven types of media tested,
Nitsch's medium showed the highest rate of somatic embryogenesis. Somatic embryos developed into normal plantlets and were
able to be potted.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
5.
Somatic embryogenesis was obtained from cotyledon and mature zygotic embryo callus cultures of Terminalia chebula Retz. Callus cultures of cotyledon and mature zygotic embryo were initiated on induction medium containing Murashige and Skoog (MS) nutrients with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) either 0.01 or 0.1 mg/l Kinetin and 30 g/l sucrose. Induction of somatic embryogenesis, proliferation and development was obtained through different culture passages. Embryogenic cotyledon callus with globular somatic embryos was obtained on MS basal medium supplemented with 50 g/l sucrose. Globular somatic embryos were observed from mature zygotic embryo callus on induction medium. Different stages of somatic embryo development from cotyledon and mature zygotic embryo calluses were observed on MS basal medium supplemented with 50 g/l sucrose after 4 weeks of culture. Histological studies have revealed the developmental stages of somatic embryos. A maximum of 40.3±1.45 cotyledonary somatic embryos/callus was obtained from mature zygotic embryo compared to 7.70±0.37 cotyledonary somatic embryos/callus initiated from cotyledons. Germination of somatic embryos and conversion to plants were achieved. Highest frequency of germination (46.66±0.88) of somatic embryos was obtained on MS basal medium containing benzyladenine (0.5 mg/l) with 30 g/l sucrose. 相似文献
6.
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
7.
Summary Embryogenic cultures were initiated from immature pecan zygotic embryos. Explants were induced for one week on Woody Plant Medium with either -naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid at 2, 6 or 12 mg/l, then subcultured monthly to fresh basal medium. Observations were made on callus production, embryo formation, and embryo morphology. Somatic embryo morphology and overall callus proliferation were affected by auxin type. Callus proliferation was less extensive and more somatic embryos resembling zygotic embryos were obtained from cultures initiated with -naphthaleneacetic acid than with 2,4-dichlorophenoxyacetic acid. Repetitive somatic embryogenesis was obtained in all auxin treatments. Conversion into plantlets was affected by somatic embryo morphology in that embryos with poorly developed apices exhibited lower percentages of conversion than those with well developed single or multiple apices. Consequently, although more embryos were obtained with 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid was the superior auxin for production of somatic embryos more likely to convert into plants.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- WPM
Woody Plant Medium (Lloyd & McCown 1980) 相似文献
8.
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
9.
Somatic embryos and embryogenic callus were initiated from immature zygotic embryos of ginseng (Panax ginseng C.A. Meyer). These somatic embryos were multiplied by adventitious (secondary and tertiary) embryogenesis and their growth and development were dependent on growth hormones in the medium. Auxins, 2,4-d, NAA, and IAA at 1.0 mg l-1 were effective in inducing secondary and tertiary somatic embryos, which proliferated directly from the apical or cotyledonary portions of the primary somatic embryos. Single somatic embryos or clusters or embryos developed from the explanted primary embryos. Cytokinin (Kn, BA) inhibited adventitious embryogenesis. Secondary somatic embryos developed to maturation and later regenerated into plantlets in two stage process; firstly elongation of the shoot axes on MS +1.0 mg l-1 Kn, secondly formation of root on 1.0 mg l-1 Kn+1.0 mg-1 GA3 medium.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- IAA
in-doleacetic acid
- Kn
kinetin
- BA
benzylaminopurine
- PSE
primary somatic embryo
- SSE
secondary somatic embryo
- TSE
tertiary somatic embryo 相似文献
10.
Callus cultures of Encephalartos cycadifolius were established from zygotic embryo explants on a modified B5 medium containing 1 mg l–1 2,4-D and 1 mg l–1 kinetin. Callus was transferred to media containing various combinations of 2,4-D and kinetin for improvement of somatic embryogenesis. Somatic embryos were produced on media with several growth regulator combinations. The somatic embryos developed from proembryos, which developed long suspensors. A dicotyledonary embryo formed at the distal end of the suspensor. The embryos turned green in light. When transferred to a medium containing 1 mg l–1 ABA the somatic embryos matured. The suspensors desiccated and these embryos rooted when transferred to a medium without phytohormones.Abbreviations ABA
abscisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
11.
Somatic embryogenesis of <Emphasis Type="Italic">Myrciaria aureana</Emphasis> (Brazilian grape tree)
Sergio Yoshimitsu Motoike Edson Santana Saraiva Marilia Contin Ventrella Crislene Viana Silva Luiz Carlos Chamhum Salomão 《Plant Cell, Tissue and Organ Culture》2007,89(1):75-81
The aim of this research was to establish a long-term somatic embryogenic cultures that could be used for cryopreservation.
For the induction of somatic embryogenesis, different levels of 2,4-D as well as the combination of 2,4-D and indole-3-acetyl-l-aspartic acid (IASP) were tested on cotyledons of zygotic embryos. The somatic embryogenic cultures were established and
maintained up to 2 years through frequent subculturing on a medium containing 2,4D + IASP. Light, activated charcoal, and
polyethylene glycol (PEG) were tested for the regeneration and maturation of somatic embryos, and the mature embryos were
germinated in JADS medium. The combination of light and PEG provided the highest number of mature embryos. The somatic embryos
obtained were smaller than zygotic embryos and lacked starch. There was an interaction between 2,4-D and IASP on the induction
and regeneration of somatic embryo in Myrciaria aureana. The combination of light and PEG increased the number of mature embryos; however, charcoal was detrimental to the process. 相似文献
12.
Sandra Correia Ana Estefânia Cunha Lígia Salgueiro Jorge M. Canhoto 《Plant Cell, Tissue and Organ Culture》2012,109(1):143-152
Somatic embryogenesis induction and somatic embryo development of the solanaceous tamarillo tree were previously established
and successfully used for plant regeneration from different explants and varieties. Somatic embryogenesis was induced in Murashige
and Skoog medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) or picloram and high sucrose concentrations (0.25 M). The
embryogenic tissues were transferred to an auxin-free medium, with reduced sucrose levels, to permit embryo development and
conversion into plantlets. This two-step protocol is often impaired by an ineffective transition from the proembryogenic masses
to embryo development. In this work, attempts to optimize the somatic embryogenesis system of tamarillo by improving the quality
of somatic embryo and embryo conversion were carried out. The results showed that the presence of a high number of abnormal
somatic embryos did not significantly inhibit plant conversion, hence indicating that shoot apical meristem development was
not affected in abnormal somatic embryos. It was also shown that the manipulation of sucrose concentration in the development
medium (0.11 M) and dark conditions before conversion increased the number of morphologically normal somatic embryos. The
comparison between mature cotyledonary zygotic and somatic embryos showed an inefficient accumulation of storage compounds,
mainly lipids, in somatic embryos. These reduced levels of lipid storage could be responsible for the abnormal patterns of
embryo development found in tamarillo somatic embryos. 相似文献
13.
Tatjana Ćosić Branka Vinterhalter Dragan Vinterhalter Nevena Mitić Aleksandar Cingel Jelena Savić Borut Bohanec Slavica Ninković 《In vitro cellular & developmental biology. Plant》2013,49(3):294-303
A simple and efficient protocol for direct somatic embryogenesis and plant regeneration of kohlrabi (Brassica oleracea var. gongylodes) was developed. Somatic embryos were induced from immature zygotic embryos at different developmental stages cultured on Murashige and Skoog medium supplemented with 0, 0.5, 1.0, or 1.5 mg/l 2,4-dichlorophenoxyacetic acid. Zygotic embryos at the early cotyledonary stage, which were cultured for 4 wk on plant growth regulator-free (PGR-free) medium, displayed the highest percentage of somatic embryogenesis (80.7%). Embryogenic tissue could be subcultured on the same medium for over 1 yr. Embryogenic lines derived from early cotyledonary stage zygotic embryos displayed the highest intensity of secondary embryogenesis (highest mean number of new somatic embryos per responsive somatic embryo explant). Histological analyses confirmed the direct origin of the secondary somatic embryos. Prolonged culturing of embryogenic tissue on PGR-free medium led to somatic embryo development into plantlets that were successfully acclimated in the greenhouse with a survival rate of 72.5%. Flow cytometry analysis showed no ploidy variation in 96.7% of the acclimated plants. 相似文献
14.
Raghavan V 《American journal of botany》2004,91(11):1743-1756
The relationship between cell expansion and cell cycling during somatic embryogenesis was studied in cultured bent-cotyledon-stage zygotic embryos of a transgenic stock of Arabidopsis thaliana harboring a cyclin 1 At:β-glucuronidase (GUS) reporter gene construct. In embryos cultured in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), following a brief period of growth by cell expansion, divisions were initiated in the procambial cells facing the adaxial side at the base of the cotyledons. Cell division activity later spread to almost the entire length of the cotyledons to form a callus on which globular and heart-shaped embryos appeared in about 10 d after culture. Anatomical and morphogenetic changes observed in cultured embryos were correlated with patterns of cell cycling by histochemical detection of GUS-expressing cells. Although early-stage somatic embryos did not develop further during their continued growth in the auxin-containing medium, maturation of embryos ensued upon their transfer to an auxin-free medium. In a small number of cultured zygotic embryos the shoot apical meristem was found to differentiate a leaf, a green tubular structure, or a somatic embryo. Contrary to the results from previous investigations, which have assigned a major role for the shoot apical meristem and cells in the axils of cotyledons in the development of somatic embryos on cultured zygotic embryos of A. thaliana, the present work shows that somatic embryos originate almost exclusively on the callus formed on the cotyledons. Other observations such as the induction of somatic embryos on cultured cotyledons and the inability of the embryo axis (consisting of the root, hypocotyl, and shoot apical meristem without the cotyledons) to form somatic embryos, reaffirm the important role of the cotyledons in somatic embryogenesis in this plant. 相似文献
15.
We elucidated the relationship between cell proliferation and somatic embryogenesis in the culture of carrot cotyledons. Fresh
weights of the cotyledon expiants were determined every five days while being cultured on a medium containing 2,4-D. Callus
production increased exponentially from Day 20 to Day 25, showing a two-fold rate of proliferation. To examine the embryogenic
potential of the callus, we pre-cultured cotyledon explants on an MS medium with 2,4-D, then transferred them to an MS basal
medium at five-day intervals. Somatic embryos formed most frequently when the cotyledons were pre-cultured for 20 days on
an MS medium that contained 5 μ2,4-D. The frequency of somatic embryo formation was 81%, while that of normal embryos with
two cotyledons was 51% among those formed on a hormone-free medium. We used FACScan analysis to relate the embryogenic potential
of the callus to the S phase in the cell cycle of cultured cells. The S phase was high after 25 days of culture on the medium
with 5 μM 2,4-D. In contrast, the frequency of normal embryogenesis was higher at Day 20 of the pre-culture period. Culturing
embryogenic calli on a medium with 5 μM 2,4-D was most favorable for producing somatic embryos with two cotyledons. We verified
that active somatic embryogenesis was apparently related to cell division activity; somatic embryos induced from actively
dividing cells were apt to accompany cotyledonary abnormality. 相似文献
16.
Maggie Panaia Eric Bunn Jen McComb 《In vitro cellular & developmental biology. Plant》2011,47(3):379-386
Somatic embryogenesis was developed as a method of mass propagation for Lepidosperma drummondii (Cyperaceae), a difficult to propagate but important species for post-mining restoration in a region of high plant biodiversity,
in the southwest of Western Australia. Cultures were initiated from excised zygotic embryos, shoot cultures to rhizomes. Only
zygotic embryos of L. drummondii developed somatic embryos, with half strength Murashige and Skoog basal medium (BM) and 1 μM 2,4-dichlorophenoxyacetic acid
(2,4-D) being the most effective combination. The first culture cycle yielded a mean of 30 somatic embryos per excised zygotic
embryo forming an embryo cluster. After a further 6 wk in culture (on fresh BM with 1 μM 2,4-D), approximately 350 somatic
embryos per starting embryo cluster were recorded. Following regular sub-culturing of primary somatic embryo clusters onto
fresh media (every 4 wk), more than 74,000 secondary somatic embryos were estimated to have been produced after eight subculture
periods. This translates to between 1,000 and 2,000 somatic embryos produced from an estimated 45 mg of starting tissue per
culture plate or potentially 22,0000–44,000 somatic embryos per gram of tissue. This is a significant improvement over all
previous methods used to propagate L. drummondii, in which typical in vitro shoot multiplication rates are as low as 1.43 per 8 wk. This also compared favourably with published data and concurrent
experiments undertaken in this study (as an extra control measure) on somatic embryo production for a related species Baloskion tetraphyllum (using the same BM with 1 μM 2,4-D and coleoptile segments as explants). Various media combinations were investigated for
efficacy in converting somatic embryos into plants with best results ranging from 86% to 100% conversion for B. tetraphyllum on BM without plant growth regulators. Development of L. drummondii somatic embryos into plants was not observed on BM without plant growth regulators. However, a best result of 39% conversion
to plants was observed on BM with 1 μM thidiazuron. This is the first report of successful development of somatic embryogenesis
and conversion of somatic embryos into plants using thidiazuron for the Australian cyperale L. drummondii. 相似文献
17.
The somatic embryogenesis potential of zygotic embryo axes of peanut (Arachis hypogaea L. cv. DRG-12) at different stages of development was evaluated by culturing on MS medium with 18.1 μM 2,4-dichlorophenoxyacetic acid (2,4-D). A 100 % frequency with 18.3 somatic embryos per explant was observed from 4 mm long immature zygotic embryo axes collected 31 – 40 d after pollination. Medium supplemented with 16.6 μM picloram resulted in slow development of somatic embryos whereas in the presence of 21.5 μM α-naphthaleneacetic acid (NAA), the explants underwent maturation with induction of roots after 30 d. The changes in protein profiles in zygotic embryo axes at different stages of development correlated with their potential to form somatic embryos. Immature zygotic embryo axes exhibited high frequency somatic embryogenesis in the stage preceding abundant accumulation of 22 and 65 kDa proteins. The content of 22 and 65 kDa proteins decreased immediately after culture on medium fortified with 18.1 μM 2,4-D and increased again after 12 d of culture coinciding with the development of somatic embryos on the explants. The content of 22 and 65 kDa proteins was low at 15 d of culture on medium supplemented with 16.6 μM picloram possibly due to slow development of the somatic embryos on the explant. On maturation medium containing 21.5 μM NAA, a marked increase in the content of 22 and 65 kDa proteins in 15 d-old cultures was observed. 相似文献
18.
Purnima Tyagi Shruti Khanduja S. L. Kothari 《Journal of plant biochemistry and biotechnology.》2005,14(2):197-200
An efficient protocol for direct somatic embryogenesis in Capparis decidua has been developed. Mature zygotic embryos cultured in Murashige and Skoog (MS) liquid medium with 2,4-0 (0.1 mg l?1) and BA (0.5 mg l?1) produced somatic embryos directly without an intervening callus phase from the subepidermal cells. Treatment with ABA promoted maturation of somatic embryos and BA (1 mg l?1)promoted germination. One zygotic embryo produced approximately 230 somatic embryos within 14 to 15 weeks. Embryos germinated in eight weeks and acclimatized plants were transferred to pots. 相似文献
19.
云南大叶茶体细胞胚发生及体细胞胚苗形成体系的建立 总被引:9,自引:0,他引:9
利用云南大叶茶(Camellia sinensis var.assamica Kitamura)胚性细胞系(CL_1)中悬浮培养物,建立了高频率同步化体细胞胚发生及体胚苗形成体系。以改良的MS为基本培养基,将CL_1中培养物由液体保持培养基(0.1mg/L 2,4-D 0.5mg/L 6-BA)继代转入液体诱导培养基(0.05mg/L 2,4-D 0.50mg/L6-BA),暗培养诱导28d,转入不含任何激素的液体分化培养基中再培养28d,获得了不同发育时期的体细胞胚,其发生频率为81.5%。不同发育时期的体细胞胚用不同目的细胞筛收集,在液体生长培养基(1/2 MS 1.0mg/L GA_3 0.5mg/L 6-BA)中培养发育成熟。ABA有利于高质量体细胞胚的形成。20~70月大小的体细胞胚在固体生长培养基中成苗转换率为75%。在液体悬浮培养条件下观察记录了体细胞胚发育过程,证实其过程与合子胚的形态发生过程相似。 相似文献
20.
The system of high synchronous frequency of somatic embryogenesis and somatic embryo seedling formation was established by means of embryonic cell hne 1 ( CL1 ) of Camellia sinensis var. assamica Kitamura. Modified MS was used as the basic medium. Cultures of CL1 was transferred to the aqueous induced medium (0.05 mg/L 2,4-D + 0.50 mg/L 6-BA) from the maintenance medium (0.1 mg/L 2,4-D + 0.5 mg/L 6-BA) for somatic embryos induction under dark condition. 28 days later, they were cultured in the liquid differentiation medium. Various kinds of somatic embryos were obtained after another 28 days. The frequency of somatic embryos was 81.5 %. Various mesh sizes of sieves were applied to collect the somatic embryos in different developmental stages which could develop to mature stage in the aqueous growth medium ( 1/2 MS + 1.0 mg/L GA3 + 0.5 mg/L 6-BA). ABA was effective to promote the formation of highly qualified somatic embryo. The mature somatic embryos sized 20 to 70 mesh had the conversion frequency 75 %. The development of somatic embryogenesis studied under a cell suspension culture system was similar to the zygotic embryogenesis. 相似文献