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1.
In vitro gynogenesis in Beta vulgaris L.: Effects of plant growth regulators,temperature, genotypes and season 总被引:1,自引:0,他引:1
Marylise Doctrinal Rajbir S. Sangwan Brigitte S. Sangwan-Norreel 《Plant Cell, Tissue and Organ Culture》1989,17(1):1-12
In order to increase the yield of haploid sugarbeet plants obtained from in vitro culture of unfertilized ovules, we have studied several factors: (1) the nature and concentration of various growth hormones, (2) the temperature for the cultured ovules, (3) genotypes and (4) seasonal effects. We found N6 medium [7] supplemented with 2.85 M of indoleacetic acid (IAA) and 0.88 M of 6-benzylaminopurine (BAP) or N6 medium containing 2.3 M of kinetin (KIN) to give the best qualitative and quantitative results for the culture of ovules. A two-step process for the induction and the development of gynogenic embryos was used. For the initiation of the embryos from the cultured ovules, the temperature of 27°C was favoured to that of 24°C. A pronounced and very significant seasonal effect was observed; the month of June was favoured to that of the other summer months tested. The effect of genotype was significant only for certain media. Using the above concentrations and combinations, a yield of viable gynogenic plants (plants/100 cultured ovules) between 6 and 10% (depending on the genotype) was obtained. In each case, 81% of these were haploids. 相似文献
2.
Desana Li >sková José Ruiz Ordóñez Alexander Lux Alfredo Piñeyro López 《Plant Cell, Tissue and Organ Culture》1994,36(3):339-343
Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10-2 mol l–1, gibberellic acid (GA3) 3.10-2 mol l–1, and 6-benzylaminopurine (BA) 2 mol l–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l–1, GA3 14 mol l–1, and kinetin 5 mol l–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l–1 and BA 10 mol l–1 was induced.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- NAA
1-naphthaleneacetic acid
- TEM
transmission electron microscopy 相似文献
3.
Somatic embryogenesis from mature elm (Ulmus minor Mill.) in vitro-cloned material is possible. Embryogenic callus was obtained from leaves inoculated on two different MS-based media—one supplemented with 2.3 M 2,4-dichlorophenoxyacetic acid (I2) and the other supplemented with 1.1 M kinetin (I6). However, only leaves cultured on medium I6 produced somatic embryos, at the globular stage, when embryogenic callus was maintained in induction media. When embryogenic callus from medium I6 was transferred to basal medium, somatic embryos with green cotyledons were obtained. An average of 35.9% of these embryos converted easily into normal plants in conversion medium with 1% sucrose. Acclimatisation reached 39.7%, and this was not significantly different from a control group consisting of plants propagated by axillary buds. No morphological differences were observed between plants derived from somatic embryos and control plants. Also, no differences in ploidy were detected between the somatic embryo-derived plants and the mother plants.Abbreviations
BA:
Benzyladenine
-
C1, C2:
Conversion media
-
2,4-D:
2,4-Dichlorophenoxyacetic acid
-
Kn:
Kinetin
-
NAA:
-Naphthaleneacetic acid
-
PI:
Propidium iodide
-
I2, I6:
Induction media
Communicated by D. Bartels 相似文献
4.
Shubhada Thengane S. V. Paranjpe S. S. Khuspe A. F. Mascarenhas 《Plant Cell, Tissue and Organ Culture》1986,6(3):209-219
An interspecific hybrid of the sexually incompatible species G. hirsutum cv. Laxmi and G. arboreum cv. Jyoti was obtained through in ovulo embryo culture. Eightto twelve-day-old ovules were excised and cultured on Beasley and Ting's medium supplemented with Indol-3 acetic acid (5×10-6 to 7×10-6 M), Kinetin (5×10-6 to 5×10-8 M), Gibberellic acid (5×10-7 to 5×10-9M), Ammonium chloride (5 to 15mM) and Casein hydrolysate (50 to 200mg/l) added individually and in various combinations along with sucrose. No single medium was adequate to ensure complete development of the fertilized ovules to plantlets, thus necessitating a sequential five step transfer to different media. Cytological studies confirmed the hybrid nature of the plants.Abbreviation IAA
Indol-3 acetic acid
- Kn
Kinetin
- GA3
Gibberellic acid
- CH
Casein hydrolysate
- NAA
-Naphthalene-acetic acid
- BT
Beasley and Ting's basal medium
- MS
Murashige and Skoog's basal medium
- W
White's basal medium
NCL Communication number 3823. 相似文献
5.
Somatic embryos of Washington Navel sweet orange (Citrus sinensis (L.) Osb.) derived from in vitro cultured ovules excised from immature fruits, were frozen to the temperature of liquid nitrogen. A method of slow cooling at a rate of 0.5°C min-1 down to –42°C followed by storage in liquid nitrogen was used. Thawing was achieved by keeping the specimens at room temperature for 15 min. A small number of frozen embryos survived and developed into proliferating cultures that produced whole plants. The plants obtained from frozen cultures were transferred to soil and are growing successfully. 相似文献
6.
Polyethylene glycol and abscisic acid improve maturation and regeneration of<Emphasis Type="Italic"> Panax ginseng</Emphasis> somatic embryos 总被引:2,自引:0,他引:2
Embryogenic culture was initiated from mature zygotic embryos of Panax ginseng. Multiple somatic embryos formed and proliferated on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2.26 M) and kinetin (0.046 M). Mature as well as immature somatic embryos grew into plantlets lacking roots on the same media. Histomorphological analysis of somatic embryos treated with abscisic acid (ABA) and polyethylene glycol (PEG 4000) showed a slight improvement in the root meristem organization of torpedo-stage embryos (embryos were more compact and their cells exhibited a lower degree of vacuolation). Shoot regeneration of non-treated somatic embryos was 31% while that for somatic embryos treated with PEG 4000 and ABA was 70%. Moreover, 75% of plants regenerated from PEG- and ABA-treated embryos formed roots while plants from non-treated embryos did not form roots.Abbreviations
ABA
(±)-Abscisic acid
-
BAP
N
6-Benzyladenine
-
2,4-D
2,4-Dichlorophenoxyacetic acid
-
GA
3
Gibberellic acid
-
Kin
Kinetin
-
MS
Murashige and Skoog medium
-
PEG 4000
Polyethylene glycol 4000
-
PGR
Plant growth regulators
Communicated by H. van Onckelen 相似文献
7.
Suk W. Kim Nam H. Song Kyung H. Jung Sang S. Kwak Jang R. Liu 《Plant cell reports》1994,13(6):319-322
Summary A system for high frequency plant regeneration from cell suspension cultures in Catharanthus roseus is described. Calli were obtained from anthers cultured on Murashige and Skoog's medium supplemented with 1 mgl-1
-naphthaleneacetic acid and 0.1 mgl-1 kinetin. After the second subculture on solid medium, embryogenic callus was identified and transferred to liquid medium to initiate suspension cultures. Cells dispersed finely in the medium were subcultured at 14-day intervals. Upon plating onto the basal medium, yellowish compact colonies proliferated from the cells and more than 80% of them gave rise to somatic embryos. Subsequently, plantlets developed from the embryos. Both the plantlets and the source plants showed the normal somatic chromosome number of 2n=2x=16.Abbreviations MS
Murashige and Skoog
- MSNK
MS medium + 1 mgl-1 NAA + 0.1 mgl-1 kinetin
- NAA
-naphthaleneacetic acid 相似文献
8.
Somatic embryogenesis was obtained from mature cassava cotyledons explants. A two-step medium sequence was developed for efficient embryogenesis. Application of 2,4-D (4 mg l-1) yielded proembryogenic masses which developed into somatic embryos after transfer to a medium containing NAA (0.01 mg l-1), BA (0.1 mg l-1) and GA3 (0.1 mg l-1). The 2,4-D concentrations used for embryo initiation strongly influenced embryo development. Among the cultivars tested, TMS 30395 was most responsive. Full strength MS basal medium alone or with 4 x MS micro salts was efficient for the formation of somatic embryos. Casein hydrolysate, adenine sulfate, nicotinic acid, glycine, tryptophan, and serine were ineffective for embryo development. High sucrose concentration (6%, w/v) inhibited the induction of somatic embryos, while 6% sucrose was optimal concentration for the development of somatic embryos after an induction treatment using 2% sucrose. Addition of 0.52 mg l-1 ABA to the induction media resulted in an increase in somatic embryos production. The ploidy levels of the regenerated plantlets were determined by flow cytometry analysis. Fifty regenerants tested were all tetraploids as the source plants and were morphologically normal. The implications of these results are discussed in relation to genetic transformation using the cotyledons as the explant source.Abbreviations ABA
abscisic acid
- BA
6-benzylaminopurine
- DAPI
4,6-diamidino-2-phenylindole
- SR
101 sulforhodamine
- GA3
gibberellic acid
- MCPA
methyl- chlorophenoxyacetic acid
- NAA
naphthalen-acetic acid
- PCPA
P-chlorophenoxyacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- 2,4,5 T
2,4,5-trichlorophenoxyacetic acid 相似文献
9.
Shoot tips and leafy bud fragments removed from offshoots of adult date palms (Phoenix dactylifera L.) were cultured on a nutrient medium containing the Murashige and Skoog inorganic salts, 453 M 2,4-dichlorophenoxyacetic acid, 14.8 M N6-(2-isopentenyl)adenine and 3 g l-1 activated charcoal to develop nodular callus after 8 months of culture. Callus was cultured in agar-solidified and stationary or shaken liquid media containing half-strength MS inorganic salts, 3 g l-1 activated charcoal and different sucrose concentrations to study the influence of these factors on somatic embryogenesis. The best conditions for embryo development were culturing in liquid medium shaken at 100 rpm for a period of 2 weeks without sucrose, followed by culture on 3% sucrose.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- 2iP
N6-(2-isopentenyl) adenine
- MS
Murashige & Skoog (1962)
- rpm
revolutions per minute 相似文献
10.
Synchronization of somatic embryogenesis at high frequency is a useful system for the mass production of embryos. Many attempts have been carried out, however, it was difficult to obtain the system in which most of the initial embryogenic cells or cell clusters synchronously differentiate to embryos. In carrot suspension cultures, high frequency, synchronous embryogenesis systems (following three systems) have been established.(1) Small spherical single cells from suspension cultures obtained by sieving and density gradient centrifugation in Percoll solutions differentiated to embryogenic cell clusters at high frequency when they were cultured in a medium containing 2,4-dichlorophenoxyacetic acid (0.05 micromolar), zeatin (1 micromolar) and mannitol (0.2 molar). (2) Embryogenic cell clusters from suspension cultures obtained by sieving, density gradient centrifugation in Ficoll solutions, and subsequent centrifugation at a low speed for a short time synchronously differentiated to embryos, especially globular embryos at high frequency, when they were cultured in a medium containing zeatin (0.1 micromolar) but no auxin. (3) Embryogenic cell clusters obtained by above method are cultured at cell densities of 2×103 cell clusters ml-1. Globular embryos which were sieved from embryos induced synchronously differentiated to torpedo-shaped embryos at high frequency when they were cultured at densities below 150 globular embryos ml-1.Using these systems, the whole process of embryogenesis from single cells to whole plants could be synchronously induced at high frequency.Abbreviations ABA
abscissic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- GA3
gibberellin A3
- IAA
indoleacetic acid
- NAA
naphthylacetic acid 相似文献
11.
Hypocotyl segments of 2- to 3-week-old radish (Raphanus sativus L. cv. F1 Handsome Fall) seedlings produced yellowish compact calli when cultured on Murashige and Skoog's (MS) medium supplemented with 1 mgl-1 2,4-dichlorophenoxyacetic acid (2,4-D). Upon transfer onto medium containing 6-benzyladenine and -naphthaleneacetic acid, up to 5.3% of the calli gave rise to a few somatic embryos. When subcultured for 3 to 6 months, 7% of the yellowish, compact calli produced white, compact calli which formed numerous embryos. These calli maintained their embryogenic capacity for over 18 months. When cultured on medium containing 0.1 to 3 mgl-1 2,4-D, up to 90% of longitudinally sliced somatic embryo halves produced calli with numerous secondary embryos. Embryos were transferred onto medium containing 0.1 mgl-1 2,4-D and 1 mgl-1 abscisic acid where they developed into the cotyledonary stage. Upon transfer onto half-strength MS basal medium, approximately 90% of the embryos developed into plantlets. These plantlets were successfully transplanted in potting soil and after cold treatment they were grown to maturity in a phytotron.Abbreviation 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzyladenine
- GA3
gibberellin A3
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog
- NAA
-naphthaleneacetic acid 相似文献
12.
Somatic embryogenesis and plant regeneration from immature embryos of saw palmetto,an important landscape and medicinal plant 总被引:1,自引:0,他引:1
Somatic embryogenesis and plant regeneration from immature zygotic embryos was achieved for saw palmetto (Serenoa repens (Bartr.) Small). Embryos, isolated from immature fruit of native-grown plants, were cultured on Murashige and Skoog medium plus 0.15% (w/v) activated charcoal and supplemented with 452 M 2,4-dichlorophenoxyacetic acid (2,4-D) and 14.7 M N6-(2-isopentenyl)adenine (2iP). Clusters of somatic embryos developed from all immature zygotic embryos 5 weeks after culture initiation. After 12 weeks, explants were transferred to the same medium with the amount of 2,4-D reduced to 90.4 M which resulted in somatic embryo proliferation. Somatic embryos were then transferred to the basal medium containing 0.9, 9 M thidiazuron (TDZ), or no growth regulator for conversion into plantlets. The 9 M TDZ treatment was ineffective for plant regeneration. However, 12% of the embryos subcultured on 0.9 M TDZ were able to produce complete plantlets. Shoot production was obtained from 35% of the embryos subcultured in the absence of growth regulators. Rooting (100%) was achieved when these shoots were transferred onto medium containing 22.2 M -naphthaleneacetic acid (NAA). 相似文献
13.
Immature zygotic embryos of two wheat (Triticum aestivum L.) genotypes, known for their different ability to generate embryogenic callus, were used as initial explants to establish callus cultures. Embryogenic and non-embryogenic calluses were obtained from the competent genotype (`Combi'), while only non-embryogenic callus was produced by the incompetent one (`Devon'). The morphogenetic competence of each callus type was evaluated by transferring some segments to regeneration conditions. The endogenous hormone concentrations (free indole-3-acetic acid [IAA], abscisic acid [ABA], gibberellins 1, 3 and 20 [GAs], zeatin/zeatin riboside [Z/ZR] and N
6[2-isopentenyl] adenine/ N
6[2-isopentenyl] adenosine; [iP/iPA]) of the initial explants were determined by means of radio-immunoassay and showed that the only difference was the higher concentration of ABA found in the embryos of the most competent genotype; whose embryos showed a reduced rate of precocious germination. When analysing the endogenous hormone concentrations in the various callus types generated in each genotype, it was found that only differences in the free IAA concentrations were associated with variations in the morphogenic properties of the calluses. Higher concentrations of endogenous free IAA were typical of embryogenic callus cultures. It was also observed that a loss in the embryogenic competence of the calluses, due to a prolonged time of culture, occurred concomitantly with a reduction in free IAA concentrations, practically to the concentrations found in the non-embryogenic calluses. 相似文献
14.
Somatic embryogenesis was achieved from mid-rib and internodal calluses of Mussaenda erythrophylla L. cvs. Queen Sirikit and Rosea cultured on Murashige and Skoog basal medium containing 8.9 M BA+0.57 M IAA+10 mg l-1 ascorbic acid. Clumps of somatic embryos were separated and grown into complete plantlets when transferred to 1/2 MS medium+37 M adenine sulphate with 2% (w/v) sucrose. 相似文献
15.
Exogenous plant growth regulators are known to increase the efficiency of interspecific and intergeneric crosses. In vitro floret culture provides a defined system for assessing the importance of various plant growth regulators on the determinants of haploid production efficiency (seed set, embryos per seeds, and plants per embryos) in Hordeum vulgare × Hordeum bulbosum crosses. The individual and combined effects of three plant growth regulators (2,4-D, GA3 and kinetin) on in vitro seed growth, embryo development and haploid production efficiency were tested in floret culture of the cross H. vulgare, cultivar Klages × H. bulbosum. All treatments, except kinetin alone, produced larger seeds and more embryos/100 seeds than the control (no plant growth regulator). 2,4-D alone was superior to GA3 alone in haploid production efficiency (70.6 vs. 51.5) as measured by the number of plants regenerated/100 florets pollinated. Although kinetin +2,4-D+GA3 produced the largest seeds and embryos, no advantage over 2,4-D alone was observed in haploid production efficiency. 2,4-D alone or kinetin +2,4-D are recommended for the purpose of barley haploid production in floret culture using the bulbosum method.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid 相似文献
16.
P. K. Gupta S. V. Kendurkar V. M. Kulkarni M. V. Shirgurkar A. F. Mascarenhas 《Plant cell reports》1984,3(6):222-225
Complete plants were grown from zygotic embryos cultured on Y3 basal liquid medium supplemented with coconut milk, BA and NAA. Explants from stem, leaf and rachilla of mature coconut trees turned green and swelled on Y3 semi-solid basal media supplemented with 2,4-D, K, NAA, BA and activated charcoal. Callus was initiated in explants from the subapical regions of the stem on Y3 basal medium supplemented with 2,4-D (4.52×102M). Globular embryo-like structures were obtained when this callus was subcultured to auxinless medium. Root formation was obtained from leaf explants on Y3 basal medium containing citric acid, ascorbic acid and 2,4-D (4.52×102 M). Globular embryo-like structures were also obtained directly from leaf explants on a Y3 basal medium supplemented with 2,4-D (2.26×102 M). Callus isolated from rachilla explants on Y3 basal medium containing 2,4-D(4.52×102 M), formed nodular structures when transferred to medium with 2,4-D (2.3×101 M). These nodules developed roots from the base of the nodular growth whereas from the upper portion shoots were observed on Y3 basal liquid medium.Abbreviations K
kinetin
- BA
Benzyl adenine
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- NAA
Naphthalene acetic acid
- CM
Coconut milk
- IAA
Indole acetic acid
- 2iP
N6-r-r-dimethyl allyl amino purine
NCL Communication No. 3471 相似文献
17.
José Nobre 《Plant Cell, Tissue and Organ Culture》1996,46(2):151-155
In vitro clonal propagation of native Mediterranean Lavandula stoechas has been achieved from mature field-grown plants. Procedures have been developed for reducing shoot hyperhydricity during in vitro culture establishment and shoot multiplication stages. Shoot multiplication was obtained, in 4–5 weeks, from single node explants cultured on a basal medium containing Margara N30K macrosalts and supplemented with 217.2 M adenine hemisulphate (AdS) and 0.05 M NAA. In vitro rooting (100%) of the shoots was observed on basal medium containing 5.4 M NAA.Abbreviations BA
N6-benzyladenine
- AdS
adenine hemi-sulphate
- GA3
gibberellic acid
- NAA
1-naphthaleneacetic acid
- PVP
polyvinylpyrrolidone 相似文献
18.
A yield of 2–4×106 protoplasts/g F.W. could be obtained when fresh cauliflower inflorescence segments were digested with 2% cellulase Onozuka R-10, 1% cellulase RS and 0.4% Macerozyme R-10 in CPW18S for 7 to 10 h. Purified protoplasts were cultured in K8p liquid and agarose medium. Although protoplasts in liquid medium divided earlier than in agarose, protoplast-derived cells in liquid culture could not avoid browning. With agarose culture, sustained division and callus formation could be achieved. After 20 days, calli were transferred onto B5 agar medium with ZT 1.5 mg l-1, BA 0.5 mg l-1 and IAA 0.1 mg l-1 for shoot formation. The frequency of bud formation varied from 56.7% for calli of 1mm in size to 5.6% for 5mm calli. The shoots formed were rooted in B5 medium containing 0.5 mg l-1 IBA, and the regenerated plants were transplanted to pots and grew normally. It took about two months from protoplasts to the regenerated plants.Abbreviations Ade
adenine
- BA
6-benzyl aminopurine
- CH
casein hydrolysate
- CM
coconut milk
- 2,4-D
2,4,-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- Gln
glutamine
- NAA
-naphthylacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- ZT
zeatin 相似文献
19.
The effects of sucrose on maturation and of plant growth regulators on germination of soybean somatic embryos were investigated for the purpose of developing an efficient culture method for plant recovery. Somatic embryos produced on medium with a low sucrose concentration (5 gl-1), less than 1 mm in length, 0.6 mg in fresh weight, and green in color, were grown for 2 weeks on MS medium containing 5 gl-1 or 30 gl-1 sucrose and then for another 5 weeks on MS medium containing 5–90 gl-1 sucrose. The highest increase in fresh weight of somatic embryos was obtained in the treatment of transferring from 30 gl-1 sucrose (2 weeks) to 60 gl-1 (5 weeks). With the increase in fresh weight, the somatic embryos gradually changed color from green to yellow, and finally to white, when they stopped growth. Soybean seed storage proteins (-conglycinin and glycinin) were accumulated in somatic embryos under tissue specific and stage specific control analogous to that in zygotic embryos. Exogenous gibberellic acid was effective in promoting precocious germination of premature soybean somatic embryos, but was not necessary for the germination of mature somatic embryos. The efficiency of somatic embryo germination was as high as 77% from semi-wild soybean and 60–64% from cultivated soybeans, showing that the plant regeneration system developed in this study was efficient and practical.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BR
brassinolide
- GA3
gibberellic acid
- IBA
indolebutyric acid
- NAA
-naphthaleneacetic acid
- PAGE
polyacrylamide gel electrophoresis
- SDS
Sodium Lauryl Sulfate 相似文献
20.
Haploid plants were regenerated from cultured unfertilized ovaries of Hordeum vulgare L. (barley). Optimal response was obtained by the addition of 0.6 M 4-chloro-2-methylphenoxyacetic acid (MCPA), 2.8 M indole-3-acetic acid (IAA) and 4.4 M 6-benzyladenine (BA) in the N6 medium. Further increase in the rate of callus formation and the number of green plants produced was possible with the addition of 90 g/l sucrose and 100 g/l coconut water. The stage of development of the ovaries at the time of culture was critical; the largest number of plants being produced by ovaries from flowers at the trinucleate stage of pollen.Abbreviations (BA)
6-benzyladenine
- (MCPA)
4-chloro-2-methylphenoxyaceticacid
- (2,4-D)
2,4-dichlorophenoxyaceticacid
- (GA3)
gibberellic acid
- (IAA)
indole-3-acetic acid 相似文献