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1.
Callus cultures were established from the scutellum, scutellar node and radicle region of immature embryos of rye and octoploid
triticale on modified Murashige-Skoog basal medium supplemented with various growth regulators. 2, 4-D, 2, 4, 5-T and 2, 4,
5-Cl, POP were found suitable for initiation and maintenance of callus cultures. Cytokinins had no or inhibitory effect on
callus induction and growth. On basal medium containing 5 mg/l of 2,4,5-Cl3 POP, 16% of triticale and 17% of rye primary cultures exhibited shoot bud regeneration after 3–4 weeks. Transfer of such
cultures to basal medium supplemented with zeatin or zeatin in combination with IAA further promoted shoot elongation and
plantlet formation. Plantlets were rooted on basal medium containing 1 mg/l NAA and were eventually transferred to soil. Chlorophyll
variants were observed in about 6% of triticale cultures. 相似文献
2.
Kim Yong Wook Youn Yang Noh Eu Rae Kim Joon Chul 《Plant Cell, Tissue and Organ Culture》1998,55(2):95-101
Embryogenic tissue was initiated using LM, LP and MS media from open-pollinated immature embryos of Larix leptolepis. The
initiation frequency varied with collection dates. The highest frequencies of embryogenic tissue initiation (60, 67 and 59%
on LM, LP and MS media, respectively) were observed from cones collected on July 30. At this time, all the excised embryos
were at the cotyledonary stage. ABA over a wide concentration and length of exposure range did not promote maturation, but
was beneficial in reducing precocious germination. Of over 400 plants regenerated, 72 were transplanted into soil mixtures
and to date, 69 of these (95%) have survived.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
3.
Summary An efficient protocol was developed to regenerate entire plants from immature embryos of elite genotypes of rye as a prerequisite
to plant transformation. Three winter genotypes and one spring genotype were tested using both immature and mature embryos
as explants. Four types of callus initiation media and five kinds of regeneration media were tested in all possible combinations.
Immature embryos gave much higher levels of plant regeneration than mature embryos, but mature embryos could be induced to
regenerate plants for all genotypes and media tested, although at low levels. A minimum stage of embryo development must be
reached before embryos can be cultured successfully. Genotypic effects were less pronounced than those reported for inbred
cereal species such as wheat and barley, but there was an effect of genotype on percentage of callus formation. There was
a significant interaction between genotype and initiation media. Composition of the initiation media affected both the percentage
of callus formation from embryos and subsequent frequencies of plant regeneration. Composition of the regeneration media had
no effect on level of plant regeneration. Immature embryos of all genotypes tested could be induced to produce 90–100% callus
on appropriate initiation media and all regenerated shoots from approximately one-half to three-quarters of the calluses produced. 相似文献
4.
Przetakiewicz A. Orczyk W. Nadolska-Orczyk A. 《Plant Cell, Tissue and Organ Culture》2003,73(3):245-256
One of the basic components of a medium influencing somatic embryogenesis of cereals from immature embryos is the type of auxin. According to some researchers, phytohormones can also play an important role during Agrobacterium-mediated transformation. In this first part of research, the influence of three types of auxins used alone or in combination of two on somatic embryogenesis and plant regeneration in three cereal species has been tested. Eight cultivars of barley, five cultivars of wheat and three cultivars of triticale have been used. Efficiency of plant development on two regeneration media, with and without growth regulators has been compared. Efficiency of regeneration characterized by frequency of explants that form embryogenic callus ranged from 25% for wheat cultivar Torka to 100% for two barley cultivars. Mean number of plantlets regenerating per explant differed significantly (from 2 to 58) depending on the type of auxin in inducing media, the type of regenerating media as well as cultivar. The biggest differences in regeneration efficiency were observed between barley cultivars, however regeneration of plants occurred in all combinations tested. The best regeneration coefficients for most barley cultivars were obtained after culture on dicamba or dicamba with 2,4-D. However, in the case of highly regenerating cv Scarlett, the most effective culture media contained picloram or 2,4-D alone. The highest values of regeneration coefficients for two triticale cultivars (Wanad and Kargo) were obtained on picloram (26.1 and 21.4, respectively) and for `Gabo' on picloram with dicamba (12.6). The range of mean number of regenerated plantlets was from 12 to 30. Dicamba alone or lower concentrations of picloram with 2,4-D were the best media influencing embryogenic callus formation in five wheat cultivars. However, the highest values of regeneration coefficients ranging from 10.6 to 26.8 were obtained at lower concentrations of picloram with 2,4-D or picloram with dicamba. R2 regeneration medium containing growth regulators was significantly better for plantlet development in several combinations (cultivar and induction medium) than the one without growth regulators. Generally, regeneration coefficients for all tested cultivars of three cereal species on the best media were high, ranging from 5.5 for barley cultivar Rodion to 51.6 for another barley cultivar Scarlett. Plantlets developed normally, flowering and setting seed. 相似文献
5.
Maria Ludovina S. Guimarães Gil S. Cruz João M. Montezuma-De-Carvalho 《Plant Cell, Tissue and Organ Culture》1988,15(2):161-167
Callus cultures with globular proembryogenic structures were induced from zygotic embryos and hypocotyl segments of Cyphomandra betacea on MS medium supplemented with 2,4-D. Proembryogenic structures produced somatic embryos and plantlets on regulator-free basal medium. Pieces of embryogenic callus subcultured on medium with the same original composition gave rise to new globular structures and the potential for plantlet regeneration has been maintained for over a year. The histological examination of these proembryogenic structures suggested that somatic embryos arise from single cells. Regenerated plants are phenotypically normal, having diploid chromosome numbers (2n = 24). 相似文献
6.
Kyung Hwan Boo Dang Viet Cao Reniel S. Pamplona Doseung Lee 《Bioscience, biotechnology, and biochemistry》2013,77(5):725-731
We established an in vitro plant regeneration system via somatic embryogenesis of Aster scaber, an important source of various biologically active phytochemicals. We examined the callus induction and embryogenic capacities of three explants, including leaves, petioles, and roots, on 25 different media containing different combinations of α-naphthalene acetic acid (NAA) and 6-benzyladenine (BA). The optimum concentrations of NAA and BA for the production of embryogenic calli were 5.0 μM and 0.05 μM, respectively. Media containing higher concentrations of auxin and cytokinin (such as 25 μM NAA and 25 μM BA) were suitable for shoot regeneration, especially for leaf-derived calli, which are the most readily available calli and are highly competent. For root induction from regenerated shoots, supplemental auxin and/or cytokinin did not improve rooting, but instead caused unwanted callus induction or retarded growth of regenerated plants. Therefore, plant growth regulator-free medium was preferable for root induction. Normal plants were successfully obtained from calli under the optimized conditions described above. This is the first report of the complete process of in vitro plant regeneration of A. scaber via somatic embryogenesis. 相似文献
7.
A. Nadolska-Orczyk A. Przetakiewicz K. Kopera A. Binka W. Orczyk 《Journal of Plant Growth Regulation》2005,24(1):2-10
Transgenic plants of triticale cv. Wanad were obtained after transformation using three combinations of strain/vectors. Two
of them were hypervirulent Agrobacterium tumefaciens strains (AGL1 and EHA101) with vectors containing bar under maize ubiquitin 1 promoter (pDM805), and both hpt under p35S and nptII under pnos (pGAH). The third one was a regular LBA4404 strain containing super-binary plasmid pTOK233 with selection genes
the same as in pGAH. The efficiency of transformation was from 0 to 16% and it was dependent on the selection factor, auxin
pretreatment, and the strain/vector combination. The highest number of transgenic plants was obtained after transformation
with LBA4404(pTOK233) and kanamycin selection. Pretreatment of explants with picloram led to the highest number of plants
obtained after transformation with both Agrobacterium/vector systems LBA4404(pTOK233) and EHA101(pGAH) and selected with kanamycin. Transgenic character of selected plants was
examined by PCR using specific primers for bar, gus, nptII, and hpt and confirmed by Southern blot hybridization analysis. There was no GUS expression in T0 transgenic plants transformed with gus under p35S. However the GUS expression was detectable in the progeny of some lines. Only 30% of 46 transgenic lines showed
Mendelian segregation of GUS expressing to GUS not expressing plants. In the remaining 70% the segregation was non-Mendelian
and the rate was much lower than 3:1. Factors that might effect expression of transgenes in allohexaploid monocot species
are discussed. 相似文献
8.
Summary A procedure for the regeneration of ‘paradise tree’ (Melia azedarach, Meliaceae) plants from immature zygotic embryos via somatic embryogenesis was developed. Somatic embryos were induced from
explants cultured on Murashige and Skoog medium supplemented with 0.45, 4.54, or 13.62 μM thidiazuron. Histological examination revealed that somatic embryos were induced directly from the explants. Further development
of somatic embryos was accomplished with Murashige and Skoog medium at quarter-strength with 3% sucrose. A large number of
plants were regenerated from somatic embryos and successfully established in soil in a greenhouse. These plants are morphologically
similar to those of seed-derived plants. This system may be beneficial for mass propagation as well as for genetic manipulation
of the ‘paradise tree’. 相似文献
9.
A protoplast to plant system in roses 总被引:7,自引:0,他引:7
Derek Matthews John Mottley Imelda Horan Andrew V. Roberts 《Plant Cell, Tissue and Organ Culture》1991,24(3):173-180
High yields of protoplasts were isolated from embryogenic suspension cultures of Rosa persica x xanthina and Rosa wichuraiana using an enzyme mixture comprising 20 g l-1 cellulase Onozuka R10, 1 g l-1 Pectolyase Y-23 and 10 g l-1 hemicellulase. Agarose-immobilized protoplasts gave the most consistent growth at a plating density of 5×104 protoplasts ml-1 on the basic medium of Kao & Michayluk (KM8p) containing 2 mg l-1 naphthaleneacetic acid and 1 mg l-1 benzylaminopurine. At 25°C in the dark, 0.004% of R. persica x xanthina protoplasts developed into colonies. Using similar culture conditions, but with a plating density of 9×104 protoplasts ml-1, 0.017% of R. wichuraiana protoplasts developed into colonies. On transfer of R. persica x xanthina colonies to Schenk & Hildebrandt's medium containing 3 mg l-1 2,4-dichlorophenoxyacetic acid, globular and later stage embryos were formed. Approximately 30% of these embryos developed into plantlets on transfer to basal Schenk & Hildebrandt's medium. Further development of the plantlets took place on cellulose plugs (Sorbarods) soaked in Murashige & Skoog's medium containing 0.05 mg l-1 naphthaleneacetic acid, 0.05 mg l-1 indole-3-butyric acid and 0.1 mg l-1 benzylaminopurine. Rose breeding is now open to the full range of in vitro genetic manipulation techniques involving protoplast technology. 相似文献
10.
In vitro somatic embryogenesis and subsequent plant regeneration was achieved in callus cultures derived from immature zygotic embryos of Acacia arabica on semi-solid Murashige and Skoog (MS) basal salts and vitamins supplemented with 8.88 MBA, 6.78 M2,4-D and 30 g l–1 (w/v) sucrose. Somatic embryos proliferated rapidly by secondary somatic embryogenesis after transfer to MS medium supplemented with 6.66 M BA, 6.78 M 2,4-D. The maximum number of somatic embryos per callus was 72.6 after 8 weeks of culture on medium containing 6.66 M BA and 6.78 M 2,4-D. The isolated somatic embryos germinated on half-strength basal MS salts and vitamins supplemented with 0.04 M BA, 0.94 M ABA and 2% (w/v) sucrose. The embryo-derived plantlets were acclimatized in the greenhouse and subsequently showed normal growth. 相似文献
11.
Lee Kang-Seop Lee Jong-Chon Soh Woong-Young 《Plant Cell, Tissue and Organ Culture》2002,68(3):241-246
An efficient method of plant regeneration from Aralia cordatasomatic embryos was developed. Somatic embryos at early stages obtained through inflorescences–derived embryogenic cell suspension cultures were matured in liquid Murashige and Skoog (MS) medium containing various concentrations of abscisic acid (ABA). For plant regeneration, mature cotyledonary embryos were transferred to solid MS basal medium for 6 weeks. Plant regeneration frequency of the embryos matured from heart-shaped embryos was proportional to the concentration of ABA from 0.76 to 3.8 M. The highest frequency (60.7%) was obtained from 3.8 M ABA pretreatment. The survival rate of the plantlets after transfer to plastic pots containing vermiculite in the growth room was 90%. All plants transferred to soil in greenhouse survived. The results indicate that micropropagation procedure can be applied for an efficient mass propagation of Aralia cordata. 相似文献
12.
S. Rueb M. Leneman R. A. Schilperoort L. A. M. Hensgens 《Plant Cell, Tissue and Organ Culture》1994,36(2):259-264
To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
3-indole-acetic acid
- BA
6-benzyladeninepurine
- S.E.M.
standard error of mean 相似文献
13.
A sunflower genotype (Helianthus annuus L. cv. Florom-328) able to regenerate plants from in vitro cultures was identified by screening hybrids and inbred lines. Protoplasts of this genotype were isolated from dark grown hypocotyls and were cultured in droplets of agarose-solidified V-KM medium covered by liquid V-KM supplemented with naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). One week later colonies were subjected to 2,4-dichlorophenoxyaceticacid for a one week period. Further culture in V-KM with reduced concentrations of NAA and BAP resulted in the appearence of somatic embryos. Maturation of embryos was achieved by culture on MS medium supplemented with NAA, BAP, gibberellic acid A3 and the ethylene inhibitor AgNO3. Embryos were then transferred onto hormone free MS medium for germination. The frequency of shoot formation in the best case was 9.6 percent of viable colonies (1.3 percent of protoplasts plated). Some of the shoots with roots could be transplanted into soil, others were grafted on hypocotyls of in vivo germinated seedlings. Eighty percent of grafted shoots and over 95 percent of rooted shoots survived. The plants flowered and produced 5 to 10 seeds each. Factors affecting the frequency of embryo formation and plant regeneration are discussed.Abbreviations BAP
6-benzylaminopurine
- GA3
gibberellic acid
- MES
morpholinoethanesulfonic acid
- MS
Murashige and Skoog medium
- NAA
naphthaleneacetic acid
- V-KM
protoplast culture medium of Binding and Nehls
- 2,4D
2,4-dichlorophenoxyacetic acid 相似文献
14.
A procedure for plant regeneration from immature seed-derived calli of rugosa rose (Rosa rugosa Thunb.) via somatic embryogenesis is described. Embryogenic calli were initiated from immature seeds 2–3 weeks after anthesis on Murashige and Skoog (MS) medium without growth regulators. Induced calli had a white, friable and nodular appearance with several proembryos. These calli were subcultured at 20-day intervals on MS medium containing 0.1–0.2 M galactose on which they grew rapidly; but somatic embryogenesis was inhibited. Somatic embryos were again induced from the subcultured calli after transferring to MS medium containing 0.1 M M fructose or sucrose but lacking growth regulators. After transferring these embryos (1–2 mm) to MS medium containing 0.1 M sorbitol, 3% of them germinated and grew into plantlets which showed sustained growth on the MS medium containing only 0.1 M sorbitol as the sole carbon source. 相似文献
15.
Abstract A regeneration protocol from leaf explants of Grindelia robusta Nutt. was developed. The combination of 0.5 mg l?1 IBA plus 0.5 mg l?1 or 1 mg l?1 BA added to Murashige-Skoog (MS) medium resulted in the best callus induction frequency; the combination of 0.4 or 0.9 mg l?1 BA plus 1.2 mg l?1 GA3 resulted in the best shoot regeneration. Rooting was successful on MS medium supplemented with 0.5 mg l?1 IBA. Hardening of G. robusta plants was accomplished in 30 days with 85% survival rate. 相似文献
16.
Mass regeneration of Coffea arabica L. somatic embryos using a temporary immersion bioreactor was improved by optimizing the immersion cycles, i.e. both the duration and the frequency of immersions. It was demonstrated that increasing the frequency of short immersions (1 min immersions every 24, 12 and 4 h) stimulated embryo production (480, 2,094 and 3,081 embryos/1-l bioreactor, respectively) and improved quality (60, 79 and 85 of torpedo shaped embryos, respectively). On the other hand, an increase in the immersion duration (1, 5 and 15 min) inhibited embryo regeneration (from 2,094 to 428 embryos per 1-l bioreactor) and negatively affected their morphological quality (from 79 to 49 torpedo-shaped embryos) and the conversion of embryos into plants (from 70 to 33). A 15 min immersion duration applied every 4 h produced hyperhydric symptoms in 90 of the embryos. Hyperhydric embryos were characterized by higher fresh weight and water content, more negative values for water potential and higher K+ content when compared to normal torpedo-shaped embryos. Micrographs showed structural problems in the globular stage, such as the existence of an irregular epidermis and an absence of reserves. Whatever the immersion cycle used, the somatic embryos exhibited water and mineral characteristics very different from those of their zygotic counterparts. The use of 1 min immersions every 4 h led to the production of the largest quantities of torpedo-shaped embryos without hyperhydricity that succeeded in regenerating plants (75 conversion). 相似文献
17.
A canopy of the traditional morphotype of Triticosecale Wittmarck and the new one were analysed in two consecutive years under different climatic conditions. On the basis of dry
matter and assimilatory surface of the plant organs during the vegetation period, the following growth indices were determined:
RGR, CGR, ULR, ULRc, LAI, LAD, LADc and HI. Radiation use efficiency was also calculated. The two possible strategies of yield
formation by analysed triticale plants were discussed.
An erratum to this article is available at . 相似文献
18.
Sara Isabel Fuentes Ramón Suárez Thelma Villegas Luis Castro Acero Georgina Hernández 《Plant Cell, Tissue and Organ Culture》1993,34(3):299-302
The in vitro embryogenic response of nine varieties of alfalfa (Medicago sativa L.) grown in México (five Mexican varieties: Puebla 76, Inia 76, Bajío 76, Sintético I and Sintético II and four foreign or introduced varieties: Moapa 69, San Joaquín II, Hairy Peruvian and Valenciana) were tested. We screened 25 genotypes from each variety in four tissue culture protocols. All the varieties, except San Joaquín II, gave a positive response in one or more of the protocols tested. The response in each variety was low; this was also observed in a wider screening performed with the varieties Moapa 69, Hairy Peruvian, Sintético I and Sintético II. Two plants from Moapa 69 were regenerated and appeared normal. 相似文献
19.
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
20.
A. Niedziela P. T. Bednarek M. Labudda D. R. Mańkowski A. Anioł 《Journal of applied genetics》2014,55(1):1-14
Triticale (x Triticosecale Wittmack) is a relatively new cereal crop. In Poland, triticale is grown on 12 % of arable land (http://www.stat.gov.pl). There is an increasing interest in its cultivation due to lowered production costs and increased adaptation to adverse environmental conditions. However, it has an insufficient tolerance to the presence of aluminum ions (Al3+) in the soil. The number of genes controlling aluminum tolerance in triticale and their chromosomal location is not known. Two F2 mapping biparental populations (MP1 and MP15) segregating for aluminum (Al) tolerance were tested with AFLP, SSR, DArT, and specific PCR markers. Genetic mapping enabled the construction of linkage groups representing chromosomes 7R, 5R and 2B. Obtained linkage groups were common for both mapping populations and mostly included the same markers. Composite interval mapping (CIM) allowed identification of a single QTL that mapped to the 7R chromosome and explained 25 % (MP1) and 36 % (MP15) of phenotypic variation. The B1, B26 and Xscm150 markers were 0.04 cM and 0.02 cM from the maximum of the LOD function in the MP1 and MP15, respectively and were highly associated with aluminum tolerance as indicated by Kruskal–Wallis nonparametric test. Moreover, the molecular markers B1, B26, Xrems1162 and Xscm92, previously associated with the Alt4 locus that encoded an aluminum-activated malate transporter (ScALMT1) that was involved in Al tolerance in rye (Secale cereale) also mapped within QTL. Biochemical analysis of plants represented MP1 and MP15 mapping populations confirmed that the QTL located on 7R chromosome in both mapping populations is responsible for Al tolerance. 相似文献