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1.
Summary Plant regeneration from cultured immature inflorescence segments of Eleusine coracana was obtained by direct shoot development and somatic embryogenesis. Direct development of shoots from cultured inflorescence segments occurred on MS medium supplemented with 2,4-D in combination with zeatin. Inflorescences with well developed spikelets differentiated at a low frequency (<5%) from callus cultures initiated on media supplemented with 2,4-D in combination with zeatin or coconut water or picloram + kinetin. Somatic embryogenesis was also induced in callus cultures growing on MS + picloram + kinetin at the end of four passages. Supplementation of the media with different concentrations of sucrose showed 3% sucrose as the best concentration for plant differentiation from somatic embryos. The majority of the regenerated plants showed the diploid chromosome constitution in their root tips. The regenerants were in general shorter with an increased number of tillers compared to the control.Abbreviations CW
Coconut water
- 2,4-D
2,4-dichloro phenoxyacetic acid
- Kn
Kinetin
- Z
Zeatin 相似文献
2.
Plant regeneration from callus cultures may provide a source ot somaclonal variation for the improvement ot the apomictic grass Poa pratensis L. It is first necessary to be able to induce callus and regenerate plants in this species at a high frequency. Variation was observed between 50 cutivars of Poa pratensis for callus induction and plant regeneration. Using the cultivars ‘Merion’ and ‘Victa’, three basal media were tested along with various media additives. Murashige and Skoog's basal medium with 0.2 mg 1?1 2,4-dichlorophenoxyacetic acid, 0.1 mg 1?1 6-benzylamanopurine, 100 mg 1?1 casein hydrolysate and 25 g 1?1 sucrose is considered to be a good medium for callus growth and plant regeneration. Embryo-like structures were observed in the callus of some cultivars but plant regeneration appeared to be predominantly from shoot meristems on the callus surface. The majority of regenerated shoots were green, but chlorophyll deficient shoots were obtained from media containing coconut milk. Green plantlets could be transferred to soil without difficulty. 相似文献
3.
Summary Twenty-five mulberry genotypes were studied for callus induction, to evaluate the effectiveness of hormones in promoting callus growth and to identify genotypes capable of regenerating plants. Fifteen genotypes showed callus initiation. Genotypic variation was also noted for longevity and rate of growth of callus cultures. Calli of different genotypes were maintained for more than one year. Frequency of callus initiation was high on Murashige & Skoog's modified medium incorporated with 2.0 mg/l 2,4-D, 100 mg/l casein acid hydrolysate and 150 ml/l coconut water. Regeneration through organogenesis was achieved in six genotypes indicating genotypic specificity. 相似文献
4.
The effects of glucose concentrations, different sugars and combinations of 2,4-D and kinetin on cell division and colony formation were examined in cultures of protoplasts isolated enzymatically from suspension cultures of Iris hollandica N6 medium supplemented with 1 mg/l 2,4-D, 1 mg/l kinetin, 200 mg/l casein hydrolysate, 250 mg/l proline, 0.3– 0.5 M glucose and 20 g/l agarose was suitable for cell division and colony formation. When colonies formed were transferred to hormone-free MS medium, many shoots were induced. In addition, when induced shoots were transferred to MS medium with 1 mg/l NAA, root induction was observed. A plant regeneration system from protoplasts of I. hollandica was thus established. 相似文献
5.
Scutellum and inflorescence explants of four genotypes of durum wheat(Triticum turgidum var. durum Desf.) were used to define culture conditions to obtain high frequencies of embryogenesis and plant regeneration in vitro. Under all conditions tested, scutellum cultures gave higher frequencies of embryogenesis and plant regeneration than inflorescence
cultures. Two different auxins, 2,4-D(2,4-dichlorophenoxyacetic acid) and picloram(4-amino-3,5,6-trichloropicolinic acid),
were compared for their effect on scutellum and inflorescence explant response in vitro. Picloram was found to significantly increase the frequency of plant regeneration from both explants. When cultures were
grown on regeneration medium containing zeatin for two three-week passages, the frequency of plant regeneration increased
by between 20–30% compared with cultures exposed to hormones for a single three-week passage. Finally, the addition of 1 mg/l
6-BAP (6-benzyl aminopurine) to the plantlet growth medium was found to enhance tiller production in regenerants. The optimized
culture conditions were applicable to the four genotypes tested and frequencies of plant regeneration varied between 97% to
100% for scutellum cultures (2 mg/l picloram in induction medium) and between45% and 80% for inflorescence cultures (4 mg/l
picloram in induction medium). The number of plants regenerated per explant was improved over previous procedures, with means
of 34 plants per scutellum, and 16 plants per inflorescence explant.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
6.
Summary With in vitro culture of ovules, interspecific hybrids have been obtained in an otherwise incompatible cross between a diploid (Gossypium arboreum) and a tetraploid (G. hirsutum) cultivated cotton. The early abortion of the embryo was prevented by repeated treatment of the flowers, immediately after pollination with a solution of gibberellic acid and naphthalene acetic acid. The ovules excised three days after pollination and cultured in a liquid medium underwent profuse proliferation, whereas on an agar-solidified medium supplemented with casein hydrolysate, indoleacetic acid and kinetin they germinated to form hybrid plants. 相似文献
7.
B. S. Ahloowalia 《Euphytica》1987,36(2):659-665
Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy. 相似文献
8.
Summary The possibility of producing agronomically-useful somaclones via organogenesis and somatic embryogenesis from callus cultures of pea (Pisum sativum L.) was studied. Organogenic calli were induced from immature leaflets on MSB medium with NAA and BAP. Embryogenic calli were derived either from immature zygotic embryos (using 2,4-D) or from shoot apices (using picloram) of aseptically-germinated seedlings.The seed progenies (T1 to T3-generation) of primary regenerants were grown in field conditions and their phenotypic variation was evaluated and compared with control, non-tissue culture-derived plant material. In addition, electrophoretic analyses of selected isoenzyme systems and total proteins have been done. The results do not show dramatic changes in qualitative and quantitative traits. The evaluation of at least two future generations (T4, T5) is planned.Abbreviations BAP
6-benzylaminopurine
- IBA
indole-3-butyric acid
- MSB
medium (mineral salts after Murashige & Skoog, 1962, vitamins after Gamborg et al., 1968)
- NAA
-naphthalene-acetic acid, picloram-4-amino-3,5,6-trichloro picolinic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- ORG
organogenesis
- SE
somatic embryogenesis 相似文献
9.
Summary Interspecific hybrids have been obtained in an otherwise incompatible cross betweenBrassica juncea × Brassica hirta through the in vitro culture of hybrid ovules and ovaries. The best response was observed from ovules and ovaries cultured 10–15 and 5–7 days after pollination respectively on a basal medium supplemented with indoleacetic acid, kinetin and casein hydrolysate. In some cases the basal cut end of the ovaries proliferated to form callus and shoots. The in vitro-derived hybrid seeds varied in their colour, size and shape, and the F1 plants in the field showed a large diversity in their morphological traits. The hybrids were sterile, and had an intermediate number of chromosomes (2n=30). 相似文献
10.
Protocols of plant regeneration have been developed for Brassica carinata for creating somaclonal variation for plant type and adaptability, so that this species can fit into cropping systems in Indian agriculture. The response of cotyledonary and stem explants was assessed for callus induction and shoot regeneration on MS and B5 basal media containing different combinations of auxin and cytokinin concentrations. MS medium supplemented with BA and NAA favoured callus induction. Supplementing MS with combinations of BA and IAA, as also with BA alone, regenerated shoots from the ex pi ants with a high frequency. The frequency of shoot regeneration and the mean number of shoots per explant were higher in cotyledons than in stem explants on identical growth regulator combinations. On B5 medium, supplemented with BA (2 mg/l) and IBA (0.4 mg/l), compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l). Genotypic differences among carinata accessions for regeneration were also observed. 相似文献
11.
Callus induced from immature inflorescences of the partially self-fertile hybrids (2 n = 35; ABDPP) between Triticum aestivum (2n = 42; AABBDD) and Agropyron desertorum (2n = 28; PPPP) led to the regeneration of 88 plants on MS medium supplemented with 2 mg/l of 2,4-D. These regenerants were used to investigate somaclonal variation and to obtain more selfed derivatives. Immature inflorescences at the stage of developing floral primordia gave the best response in terms of callus induction and plant regeneration. The regenerants exhibited great variability for most morphological traits. Although the regenerants did not exhibit variation in chromosome number, they did show a higher degree of meiotic instability than the initial hybrid. In particular, the regenerants gave much higher selfed seed-set (5.49 %) than the donor hybrid (0.46 %), so that a total of 484 selfed seeds were obtained. 相似文献
12.
A protocol for high frequency callus induction and plant regeneration from sunflower (Helianthus annuus L.) anthers is described.
Different variables using Murashige & Skoog (MS) basal medium supplemented with 2.0 mg/l α-naphthaleneacetic acid (NAA) and
1.0 mg/l N6-benzyladenine (BA) were tested for their ability to enhance the frequency of anther callusing and subsequent embryogenesis.
Of these, agar concentration, sucrose concentration, carbohydrate source had significant effect on callusing, while differences
due to incubation under dark vs light conditions, cold pretreatment of capitula for 1 to 6 days prior to anther inoculation
and genotype on callusing were non-significant. However, all these factors exerted highly significant influence on embryogenesis
when calli from the various media were transferred to medium supplemented with 0.1 mg/l NAA and 0.5 mg/l BA. With the procedure
developed, callusing as high as 100% and embryo formation at a frequency of 44% was achieved. Although complete embryos were
formed the frequency of their conversion to whole plantlets was low (14.3%). Hence, the embryogenic pathway was bypassed to
obtain multiple shoots by transferring embryogenic calli with developing embryos to MS medium supplemented with 0.5 mg/l BA.
Elongated shoots rooted on half-strength MS medium supplemented with 0.5 mg/l NAA. Cytological analysis of embryogenic callus
and somatic embryos revealed haploids at a frequency of 30% while that of rooted plants showed haploid regenerants at a frequency
of 8.3%. Nevertheless, the frequency of putative haploid plants could be enhanced through mass multiplication using nodal
explants of the regenerants.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
Summary Callus induction and green plant regeneration at high frequencies from an interspefic hybrid, Oryza sativa L. x O. rufipogon Griff. has been achieved by simply coordinating the growth regulators in the induction medium. The study was conducted with two different basal media (Potato-2 and N6) and seven different combinations of growth regulators 2,4-D, NAA and kinetin. Synergistic effects of the two auxins in enhanced anther response to callus induction and subsequent green plant regeneration were observed in both media. The highest frequency of callus induction was obtained on Potato-2 medium supplemented with 1 mg/12,4-D, 2 mg/l NAA and 1 mg/l kinetin. The same combination of growth regulators which yielded higher frequencies of callus induction also induced higher mean number of calli per anther. Although the calli formed on N6 medium showed high regenerability, there was a concomitant increase in the number of albinos among the regenerants. The auxins in the induction media had considerable influence on the regeneration capacity of the calli. The regeneration frequencies were higher from calli formed in the presence of both auxins in the induction media. The levels of growth regulator combinations seem to influence the green plant regeneration especially for calli induced on Potato-2 medium. Among the pollen grain derived plants the majority were either haploids or double haploids and very few were chromosomal variants. 相似文献
14.
Summary Leaf discs from sixBeta vulgaris lines and threeB. maritima accessions were tested for their morphogenic responses in Murashige and Skoog medium supplemented with 0.25 mg/l benzyladenine (BA). Direct shoot formation took place in two genotypes:B. vulgaris line TA33-MS andB. maritima acc. France. Rooting of differentiated shoots was achieved by culturing the shoots on a medium containing 0.2 mg/l indolebutyric acid (IBA). Our results indicate that genotypes differ in the morphogenic potential of leaf explants. 相似文献
15.
Immature embryos resulting from the cross V. vexillata × V. unguiculata were cultured on MS medium supplemented with 2,4-D
(2 mg/l) and resulted in embryogenic calli. Thirteen hybrid regenerants were obtained via organogenesis by subculturing the
calli on MS medium supplemented with BAP (2 mg/l) + adenine sulphate (40 mg/l) + CH (500 mg/l) + cowpea tender pod extract
(10%). The interspecific regenerants showed intermediate morphological traits between the parents for leaf shape, pod colour
and seed coat colour. The hybrid plants inherited stem, leaf and pod hairiness of the wild species which could serve as a
mechanical barrier against viral vectors. Electrophoretic studies of two isozyme systems, peroxidase and esterase, also confirms
the hybrid nature of the regenerants as they expressed unique bands of both parents. Cytological study of the meiotic chromosomes
revealed high frequency of univalent formation in the hybrids suggesting that the genomes of the parental species are structurally
differentiated. The hybrid regenerants exhibited high enzyme activity for three enzymes viz., peroxidase, polyphenol oxidase
and phenyl alanine ammonia lyase over the cultivated parent which may be useful in conferring resistance against viral pathogens.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
16.
Summary A high frequency plant regeneration system via organogenesis and somatic embryogenesis was established with callus cultures derived from mature zygotic embryos of different leek genotypes (Allium ampeloprasum L.). Four different callus types with varying morphogenetic potential were obtained. Relatively high concentrations of the auxin 2,4-dichlorophenoxy-acetic acid reduced callus weight and subsequent shoot regeneration and primordia formation of the callus. Shoot regeneration and primordia formation of the callus decreased after prolonged subculture on media containing 2,4-dichlorophenoxy acetic acid. A callus growth period of six weeks on Murashige and Skoog medium with 0.25–0.5 mg l-1 2,4-dichlorophenoxy acetic acid showed the highest rate of shoot regeneration after transfer of callus to regeneration medium with 1 mg l-1 kinetin.Differences between leek genotypes in callus type, callus weight, shoot regeneration and primordia formation were observed. Histological observations showed that plant regeneration took place, both via the pathway of somatic embryogenesis and organogenesis.Abbreviation 2,4-D
2,4-dichlorophenoxy acetic acid
- MS
Murashige and Skoog (1962) medium 相似文献
17.
The effect of in planta TIBA and L-proline onin vitro seedlings and cell culture of sugar beet was investigated. Sterilized seeds were grownin vitro on 1/2 MS medium supplemented with 0 or3 mg/l TIBA. Calli obtained on young leaves cultured on MS medium containing 1 mg/l
BAP, were used for the initiation of cell suspension cultures using MS basal composition supplemented with 0 or 50 mM proline.
Aliquots of 1 ml from cell suspension culture were inoculated onto the first somatic embryo induction MS medium containing
TIBA 0.5 mg/l, BAP 1.0 mg/l, and proline at 0 or 50 mM. After three weeks of culture, embryogenic calli were transferred to
the second embryo induction medium supplemented with NAA and BAP at 0.2 and 0.5 mg/l, respectively. The frequency of somatic
embryos of calli obtained from in plantaTIBA together with proline treatments on average was20 which was higher than that of the other treatments.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Plant regeneration was achieved from immature embryo-derived, calli of Pisum sativum. Embryo axes were separated from cotyledons and cultured on different media containing BAP and NAA until plantlet regeneration. Rooting of the plantlets was obtained on MS medium supplemented with 2 mg/1 IBA. Frequency of regeneration was shown to be under the influence of the genotype. Histological preparations showed de novo origin of the shoots via organogenesis. Out of 2C regenerated plantlets, 11 were diploids (2n = 14) arid 9 aneusomatic (chromosomal mosaics) with chromosome numbers ranging from 12 to 16. 相似文献
19.
Plant regeneration from protoplasts of Iris germanica L. 总被引：1，自引：0，他引：1
Summary Protoplasts were isolated enzymatically from suspension cultures derived from embryogenic calli induced by leaf base culture of Iris germanica. In protoplast culture, the effects of glucose concentration, different sugars and combinations of 2,4-D and KIN on protoplast division and colony formation were examined. N6 medium supplemented with 0.1–1 mg/l 2,4-D, 1 mg/l KIN, 200mg/l casein hydrolysate, 250 mg/l proline, 0.2 M glucose and 20 g/l agarose was suitable for protoplast division and colony formation. When colonies formed were transferred onto hormone-free MS medium, many plantlets were regenerated through somatic embryogenesis. Thus, we could establish a plant regeneration system from protoplasts of I. germanica.Contribution from the Laboratory of Plant Breeding, Faculty of Agriculture, Miyazaki University, Japan, No. 95. 相似文献
20.
Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA3, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava. 相似文献