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1.
Maira Oropeza Ana Karina Marcano Eva De García 《In vitro cellular & developmental biology. Plant》2001,37(2):211-216
Summary Previous results have shown that some proteins secreted in the culture medium are involved with the formation of embryogenic
cells and can modify somatic embryo differentiation. Undifferentiated cell suspensions grown in the presence of 13 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and obtained from embryogenic and non-embryogenic callus were used to study these
events in sugarcane plants (cv.PR-62258). The cell suspension growth curves were determined and soluble proteins were extracted
from embryogenic and non-embryogenic callus and culture medium from cell suspensions. In embryogenic callus we detected 1.43
times more protein than in non-embryogenic callus and the electrophoretic protein patterns show specific polypeptides for
both callus types. In embryogenic callus we detected a cluster of four polypeptides in the range of 38–44 kDa and another
polypeptide of 23 kDa that were not observed in non-embryogenic callus. In nonembryogenic callus there is a 35-kDa polypeptide
that was not detected in embryogenic callus. In the case of extracellular proteins, the medium from embryogenic cell suspensions
contained four polypeptides of 41, 38, 34 and 28 kDa that were slightly detected in the medium from non-embryogenic cell cultures;
we also detected a band at 15 kDa that could not be observed in the medium from non-embryogenic cell suspensions. These results
suggest that the development of embryogenic callus and cell suspensions is related to the type and amount of intracellular
proteins in the callus cells and to the secreted proteins from these cells into the medium. 相似文献
2.
Zhi-Sheng Xu Zhi-Ying Yu Meng Zhang Zhen Zhang Jian-Min Tao 《In vitro cellular & developmental biology. Plant》2014,50(2):249-256
Vitis vinifera L. cv. ‘Manicure Finger’ is one of the major table grape varieties in China. To provide a strong foundation for genetic transformation with potential for crop improvement, we undertook plant regeneration via somatic embryogenesis. Anthers and gynoecia were harvested from immature flowers and used as explants to induce embryogenic calli. Explants cultured in MS1 medium (based on Murashige and Skoog basal salts), supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.4-μM 6-benzylaminopurine (6-BA) showed the highest rates of embryogenic callus induction (3.7%?±?1.3% for anthers and 4.8%?±?2.5% for gynoecia). After several months, somatic embryos were produced from embryogenic calli cultured in plant growth regulator-free MS2 medium (with reduced sucrose). Somatic embryos (SE) at the cotyledonary stage were isolated and cultured on three different media (MS2, MS3, or B) for conversion into plantlets, the efficiency of which ranged from 63.9%?±?4.8% to 83.9%?±?8.4%. After 1 mo of in vitro culture, 80% of plants with at least six leaves were successfully transplanted into soil. SE was repeatedly induced from previously induced somatic embryos for up to 1.5 yr. Using embryogenic calli as starting material, suspension cultures containing embryogenic cell aggregates were also established in liquid MS medium supplemented with 4.5-μM 2,4-D. The embryogenic cell aggregates continued to proliferate without differentiating for successive subculture cycles. After transfer to 2,4-D-free liquid medium for 4 wk, an average of 63.7%?±?9.0% mature SEs were produced per 20 mL of liquid medium. More than 40% of somatic embryos at cotyledonary stage, derived from the suspension cultures, successfully germinated into plants using solid medium. 相似文献
3.
Jingli Yang Chenguang Zhou Lihui Liu Du Jia Zhiyang Yin Myongjo Kim Changyeon Yu Chenghao Li 《Plant Cell, Tissue and Organ Culture》2012,110(2):289-298
Embryogenic callus was obtained from young petiole, stem, and root explants of 4-year-old Siberian ginseng plants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Embryogenic callus differentiated into somatic embryos (SEs), most of which could germinate but developed abnormally. Friable embryogenic callus was induced mainly from the root regions of germinated primary SEs or regenerated plantlets on plant growth regulator-free medium. Histological studies showed that the embryogenic callus initiated from the subepidermal cells of young roots. The bioreactor system was more efficient than suspension culture regarding the number and growth of SEs, although a similar amount of embryonic tissue was used. An average of 115,370 germinated SEs developed from an initial 400?mg of embryogenic callus, and 64.7?% of germinated SEs converted into plantlets after a 4-weeks culture on agar medium. During the bioreactor culture process, secondary SEs were induced directly from SEs at various stages, a phenomenon that rarely occurred in suspension culture. These secondary SEs developed quickly and germinated during the bioreactor culture process. Proline content and peroxidase and catalase activities of SEs cultured in bioreactors were higher than in SEs cultured in suspension culture. 相似文献
4.
Compact, friable and embryogenic calli were initiated from immature inflorescences and young leaf bases of one week old seedlings of Paspalum scrobiculatum cultured on MS medium supplemented with 2,4-D. A stable, embryogenic suspension culture was initiated from these calli and maintained in a liquid version of the same MS medium. Embryogenic calli and somatic embryos were obtained by plating suspension culture cells onto semi-solid medium containing 2,4-D. Complete, normal plantlets developed on 2,4-D free medium at a high frequency from somatic embryos. NAA and BAP in the medium promoted plant development.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
Abscisic acid
- MS
Murashige and Skoog (1962)
- CM
Coconut milk 相似文献
5.
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
6.
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
7.
Extracellular proteins in embryogenic suspension cultures of Norway spruce (Picea abies) 总被引:1,自引:0,他引:1
Embryogenic cell lines of Norway spruce ( Picea abies ) varying in growth habit and morphology were compared as regards profiles of extracellular proteins. Similar proteins were detected in the culture medium by SDS PAGE and in vivo labeling experiments, indicating that the proteins were secreted. Approximately 20 protein bands could be detected in the medium of each cell line. Three of the bands represented glycosylated proteins, as revealed by Concanavalin A staining. Some of the secreted proteins were similar for all tested embryogenic lines of Norway spruce, others were either specific for a group of cell lines or for individual cell lines. A correlation was observed between the morphology of the somatic embryos in a cell line and the presence of secreted proteins. The embryogenic cell lines of Norway spruce can be divided into two main groups. A and B, where A is characterized by somatic embryos with dense embryoheads and B by somatic embryos with loosely aggregated cells in their embryoheads. When proteins secreted from a cell line belonging to group A were added to cell lines belonging to group B, the somatic embryos of the B type developed further and became more similar in morphology to A-type embryos. These observations indicate that cell lines belonging to group A secrete certain proteins to the culture medium that are essential for the development of somatic embryos of Norway spruce. 相似文献
8.
Somatic embryogenesis and plant regeneration from suspension cultures of Acanthopanax koreanum Nakai
High-frequency somatic embryogenesis was achieved from an embryogenic cell suspension culture of Acanthopanax koreanum Nakai. Stem segments were cultured on Murashige and Skoog (MS) medium containing auxins and cytokinins. Opaque and friable
embryogenic callus formed on MS medium with 4.5 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 μm kinetin or zeatin, but was highest on medium containing 4.5 μm 2,4-D alone. Embryogenic calli were transferred to MS liquid medium containing 4.5 μm 2,4-D and maintained by subculture at 2-week intervals. Initiation of somatic embryogenesis and development up to the globular
stage from embryogenic cell clumps occurred in medium containing 0.45 μm 2,4-D, whereas maturation and germination of somatic embryos occurred in MS medium lacking 2,4-D. Cytokinin treatment suppressed
the normal growth of embryos, but stimulated secondary somatic embryogenesis from the surfaces of primary embryos. Plants
from somatic embryos were acclimatized in a greenhouse.
Received: 14 January 1997 / Revision received: 17 June 1997 / Accepted: 5 July 1997 相似文献
9.
An embryogenic suspension culture of Zea mays, genotype 4C1, was obtained from friable callus that was cultured on solid medium and had been obtained from zygotic embryos. The suspension contained non-dividing elongated cells, clusters of dividing isodiametric cells, and globular, ovoid, and polar stages of somatic embryos. The single somatic embryos were blocked in shoot meristem formation: when transferred to regeneration medium they developed a root and, at the shoot side, a green cap with meristematic cells, but a scutellum and leaf primordia were not formed. In medium containing 2,4-dichlorophenoxy acetic acid, somatic embryos formed embryogenic callus aggregates, consisting of globular stage somatic embryos attached to each other via undifferentiated callus cells. These somatic embryos developed into mature embryos with the zygotic histological characteristics, such as scutellum and leaf primordia, in maturation medium, and then regenerated into plants in regeneration medium. By omitting the maturation phase, regeneration occurred via organogenesis. Polyembryos, i. e. embryos attached to each other without callus tissue in between, behaved as single somatic embryos. It is concluded that the attached callus tissue provides a factor that stimulates scutellum and leaf primordia formation.Abbreviations CMM
callus maintenance medium
- 2,4D
2,4-dichlorophenoxy acetic acid
- PCV
packed cell volume
- MS
Murashige and Skoog medium 相似文献
10.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
11.
M. Cristina Pedroso M. Salom Pais 《In vitro cellular & developmental biology. Plant》1995,31(1):31-35
Summary Two methods (I and II) for somatic embryo production from embryogenic suspension cultures ofCamellia japonica are presented. Method I, embryogenic suspension cultures, was established from suspension cultures initiated from leaf-derived
callus. These cultures were maintained by reducing agitation and increasing subculture interval. Induction of somatic embryogenesis
was achieved in MS28 medium, 6, 12, 24, and 36 mo. after culture establishment. Embryo production decreased after 1 yr of
culture. Method II, suspensions of single embryogenic cells and proembryos, was obtained from leaves cultured in liquid MS13
medium 6 wk after culture initiation. Embryo production was 23 embryos/ml. Germination of cell suspension-derived embryos
on MS56 medium was 16.7 % (±4.2%) for method I, and 35.4% (±5.1%) for method II. The embryos germinated into
plantlets with 0 to 7 axillary shoots. 相似文献
12.
Plant Regeneration from Immature Zygotic Embryo-Derived Embryogenic Calluses and Cell Suspension Cultures of Catharanthus roseus 总被引:2,自引:0,他引:2
Kim Suk Weon In Dong Su Choi Pil Son Liu Jang R. 《Plant Cell, Tissue and Organ Culture》2004,76(2):131-135
Culture conditions for plant regeneration in immature zygotic embryo-derived embryogenic cell suspension cultures of Catharanthus roseus (Madagascar periwinkle) Little Bright Eye are described. Immature zygotic embryos formed off-white, friable calluses at a frequency of 20% on Murashige and Skoog (MS) medium supplemented with 4.52 µM 2,4-dichlorophenoxyacetic acid (2,4-D) after 8 weeks of culture. After a second subculture using MS basal medium at 4-week intervals, off-white friable calluses formed a small quantity of yellowish, compact embryogenic calluses. Upon transfer to MS basal medium, embryogenic calluses gave rise to numerous somatic embryos. Cell suspension cultures were established with embryogenic calluses using liquid MS medium supplemented with 4.52 µM 2,4-D. Embryogenic cell clumps from cell suspension cultures developed into plantlets at a frequency of 56.7% when plated onto MS basal medium. Plantlets were transplanted to potting soil and grown to maturity in a growth chamber. 相似文献
13.
Somatic embryogenesis was achieved from cell suspension cultures of niger (Guizotia abyssinica Cass.). Initially, friable embryogenic calluses were induced from cotyledonary leaves of niger on Murashige and Skoog (MS) agar medium containing 5 μM 2,4-Dichlorophenoxyacetic acid (2,4-D) and 0.5 μM kinetin (KIN). Cell suspension cultures were established by using embryogenic calluses in MS liquid medium containing 5 μM 2,4-D and 0.5 μM KIN. Initiation of somatic embryogenesis and development up to globular stage from embryogenic cell clumps occurred in the liquid medium itself. Thereafter embryogenic cell aggregates were transferred to MS agar medium supplemented with 3 μM KIN for embryo differentiation, whereas maturation of somatic embryos occurred in MS agar medium containing 10 μM abscisic acid. 相似文献
14.
胡萝卜体细胞胚在人工种子制作中的分级分选 总被引:1,自引:0,他引:1
用植物胚性细胞团分选仪进行了从胡萝卜非均匀性的悬浮培养物中分级分选体细胞胚。对该仪器分选体细胞胚的操作程序及参数进行了确定,对分离的三个等级的体细胞胚的数量、大小和形态进行了测定。该仪器分选胡萝卜成熟的体细胞胚的速度为每小时从100ml 悬浮培养物中分选出872个胚。按大小次序分级分离的三个等级的体细胞胚所制作的人工种子,在无菌培养基上的萌发率分别为87.3%、75.3%和55.5%。本研究的结果表明植物胚性细胞团分选仪用来分级分选胡萝卜体细胞胚的效果是良好的,成熟胚(第一、二级)制作的人工种子比未成熟胚(第三级)制作的人工种子萌发率有显著的提高。 相似文献
15.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
16.
17.
Culture conditions for high frequency plant regeneration via somatic embryogenesis in cell suspension cultures of Chelidonium
majus var. asiaticum are described. Immature ovules formed embryogenic calluses at a frequency of 40% when cultured on Murashige
and Skoog (MS) medium supplemented with 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The optimum ovule size for embryogenic
callus formation ranged from 1 to 1.5 mm in length. Cell suspension cultures were established from embryogenic calluses using
MS liquid medium containing 4.52 μM 2,4-D. Upon plating onto MS basal medium, cell aggregates from cell suspension cultures
produced somatic embryos which then developed into plantlets. Regenerated plantlets were transplanted to potting soil and
grown to maturity in a growth chamber.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
We identified and isolated a monoclonal antibody (MAb 3G2) raised against extracellular proteins from microcluster cells of
orchard grass (Dactylis glomerata L.) embryogenic suspension culture. MAb 3G2 recognized with high specificity an antigen ionically bound within the primary
cell wall and in the culture medium of microcluster cells. Two-dimensional polyacrylamide gel analysis and blotting of proteins
on PVDF membrane showed that MAb 3G2 detected a single polypeptide of apparent molecular mass of 48 kDa and an isoelectric
point (pI) of 5.2, designated EP48. A transient expression during somatic embryogenesis was observed for EP48. Indirect immunofluorescence
showed that this protein highly accumulated in the cell walls of some single cells, microclusters and partly in proembryogenic
masses (PEMs), but not in globular embryos of the embryogenic cell line and microclusters from the non-embryogenic cell line.
Signal intensity varied between individual cells of the same population and in successive stages of somatic embryo development.
Screening of several D. glomerata L. embryogenic and non-embryogenic cell lines with MAb 3G2 indicated the presence of ECP48 in only embryogenic suspension
cultures at early stages of embryo development long before morphological changes have taken place and thus it could serve
as an early marker for embryogenic potential in D. glomerata L. suspension cultures. 相似文献
19.
Jonny Everson Scherwinski-Pereira Rodrigo da Silva Guedes Ricardo Alexandre da Silva Paulo César Poeta FerminoJr. Zanderluce Gomes Luis Elínea de Oliveira Freitas 《Plant Cell, Tissue and Organ Culture》2012,109(3):501-508
An efficient and simple system for inducing somatic embryogenesis and regenerating plantlets from immature zygotic embryos of açaí palm (Euterpe oleracea) has been developed. Embryogenic calli (ECs) were induced from immature zygotic embryos of açaí palm on Murashige and Skoog (MS) modified medium with 2,4-dichlorophenoxyacetic acid and picloram. Embryogenic frequency was dependent on auxin type and concentration. The optimal concentration of picloram for the high-frequency induction of embryogenic calli (72%) was 225 μM. ECs were then subcultured on a differentiation and maturation medium composed of MS modified medium with 2-isopentenyladenine and naphthaleneacetic acid with subcultures at 4-week intervals. SEs were converted to plants on MS modified medium with half-strength macro- and micronutrients, 20 g l?1 sucrose, and 2.5 g l?1 activated charcoal and gelled with 2.5 g l?1 Phytagel. Detailed morpho anatomical changes during the different stages of somatic embryogenesis were characterized. The development of SEs was asynchronous, and ontogenic studies confirmed that the initial cell divisions occur in the epidermal and subepidermal regions of the zygotic embryos. Broad base attachment of SEs to the epidermis indicates the presence of a suspensor. 相似文献
20.
Three methods of increasing the productivity of somatic embryogenesis in Medicago sativa L. were investigated. In the basic procedure, somatic embryos were initiated from young petioles and carried through several phases: callus formation, suspension culture, selection of the embryogenic fraction by sieving, development, maturation, desiccation and storage. The suspensions were normally separated into three fractions by sieving. Fraction I (<200 m) containing nonembryogenic cells or cell clusters was discarded. Fraction II (200–500 m) consisting of embryogenic cell clusters was collected for embryo development and maturation. Fraction III (over 500 M) containing the mixture of petiole residues with large pieces of calli and globular somatic embryos was usually discarded. Several methods to scale-up the suspension phase were unsuccessful. Direct subculture of the entire suspension by the addition of fresh liquid medium resulted in the loss of embryogenic capacity by the third subculture. Subculture of fraction II decreased embryogenic cell mass, and hence reduced total productivity. The recycling of fraction III back to fresh B5g liquid medium resulted in high productivity in the first culture but further subculture of this fraction resulted in a rapid decline in the embryogenic capacity.As an alternative, somatic embryos from the first tissue culture cycle were also used as explants for the initiation of secondary embryogenic callus. The embryogenic capacity of these somatic embryo explants declined rapidly as they matured. More than 100 secondary somatic embryos could be induced from embryo explants removed from development medium at 10 days after sieving the suspension, but only 40 somatic embryos were produced from each mature somatic embryo explant, and 13 from desiccated embryos. The secondary somatic embryos were comparable to the primary embryos in quality according to germination tests. The implications of the results to the efficiency of somatic embryo production of Medicago are discussed.Abbreviations ABA
abscisic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- DAS
days after sieving
- PPF
photosynthetic photon flux density
- SE
somatic embryo 相似文献