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1.
Callus could be induced from the cotyledon segments of Cucumis redo L. in Miller’s medium supplement with high concentration of kinetin (10–15 mg/L) or with NAA 2 mg/L plus kinetin 0.5 mg/Ll. The calli induced by NAA plus kinetin were much different from those by kinetin alone. The former was loose and soft. While the latter was tight and gradually developed new buds. Isoenzymes of glucose-6-phosphate dehydrogenases, isocitrate dehydrogenases, malate dehydrogenases and glutamate dehydrogenases were detemineted at 5, 10, 15, 20, 25 and 30 d after inoculation. The results showed that both type of calli contained four dehydrogenases during their growth period. This indicates that the pentose phosphate pathway, TCA cycle and glutamate metabolism are present in calli of both types during their growth. The isoenzymes of glucose-6-phosphate dehydrogenases and malate dehydrogenases in shoots developing calli are different from those in shootless ones. 相似文献
2.
Influence of growth regulators on bud initiation in callus of alfalfa (Medicago sativa L.) was studied by varying levels and combinations in the first medium of a two-medium sequence used to obtain whole plants. Callus of tetraploid clone S-4 (cv. ‘Saranac‘) was initiated from immature ovaries on a modified Blaydes' basal medium containing all combinations of six concentrations (0–36 μM) of kinetin (K), six concentrations (0–44 μM) of naphthaleneacetic acid (NAA), and seven concentrations (0–36 μM) of 2,4-dichlorophenoxyacetic acid (2,4-D). After 28 days the callus was challenged to form buds by transfer to the modified Blaydes' medium containing 2.0 g/liter yeast extract and 0.57 mM inositol. No buds were produced in the absence of 2,4-D in the first medium, and the frequency of bud formation on the second medium was directly proportional to the 2,4-D concentration in the range 2.3–54 μM in the preceding medium. Buds were produced in the absence of kinetin in the first medium, but its presence in the range 2.3–36 μM markedly increased bud formation. NAA was not required for bud formation, and the budding frequency increased only slightly with increasing NAA concentration in the first medium. Budding of callus of two other alfalfa clones was also influenced by the 2,4-D concentration in the initial medium. There were several indications that many of the buds were initiated on the first medium and completed development on the second medium. These included the differential effect on budding of combinations of 2,4-D, NAA and kinetin in the callus initiation medium, the specific media sequence required, and the presence of embryoids on the callus which after transfer to the yeast extract-inositol medium produced buds. 相似文献
3.
P. Profumo P. Gastaldo L. Caffaro R. M. Dameri G. Roti Michelozzi A. Bennici 《Protoplasma》1985,126(3):215-220
Summary Expiants ofCichorium intybus L. storage roots were grownin vitro on a modified Heller's medium lacking auxins and cytokinins, or supplemented with auxins (either 2,4-D or NAA) alone or with a cytokinin (kinetin) or auxin and kinetin combinations in different concentrations. The morphogenetic responses of root explants varied with the different hormonal treatments. The best response for callus growth was obtained in presence of 2,4-D. On the contrary, kinetin alone was very effective for shoot induction, increasing the formation of adventitious buds (up to 100% of the explants) in respect to control (hormone-free medium). NAA induced either shoot differentiation (in a medium frequency) or root formation. Expiants excised from root zones near to apex, which showed on hormone-free medium a very low regenerative capacity (lower than proximal zones of the root), responded to kinetin by increasing significantly the number of shoots from adventitious buds.Cytological analyses in developing primary calli showed, in all media, high incidence of amitotic phenomena confirmed by DNA cytophotometry in calli at different growth stages. The histological analysis demonstrated the formation of meristematic growth centers on the organogenesis inducing media and the subsequent development of these meristemoids as shoot (or root) apices in the callus mass.The results are discussed in comparison with previous observations of the authors inCichorium intybus (Caffaro
et al. 1982) and in relation to the action of hormonal treatments on callus formation and organogenesis. The cytological and histological results are also discussed in relation to the hormonal composition of the medium. 相似文献
4.
Shoot buds from the saplings and the fruit bearing plants of Carica papaya L.. var. Honey Dew (papaya) initially treated with Gentamycin were cultured in modified MS media, each with a different hormonal combination, for the establishment of cultures and multiplication and rooting of plants. About 43% of explants from fruit bearing plants and 69% of those from saplings remained free of contamination and retained regeneration capacity when treated in 500 mg/l Gentamycin. For the establishment of the explants a medium containing 1 mg/l GA3 and 2 mg/l kinetin was necessary. When established buds were transferred to medium containing 1 mg/l NAA and 3 mg/l kinetin, calli were initiated at cut ends of shoot buds; multiplication started on transfer to NAA (0.1 mg/l) and BAP (0.5 mg/l) medium. Cultures have been maintained for the last twenty months without any loss in multiplication rate. Rooting was induced in medium with reduced salt concentration containing 2 mg/l IBA. Shoot elongation was induced after prolonged culture in the same rooting medium.Abbreviations MS
Murashige and Skoog, 1962
- SH
Schenk and Hildebrandt, 1972
- GA3
Gibberellic acid
- Kn
Kinetin
- NAA
Napthaleneacetic acid
- BAP
6 -Benzylaminopurine
- IBA
Indole-3-butyric acid
- IAA
Indole-3-acetic acid 相似文献
5.
濒危中药材太白米组织培养及总黄酮含量测定 总被引:1,自引:0,他引:1
以太白米(Notholirion bulbuliferum)地下小鳞茎为外植体,通过在MS基本培养基添加不同浓度2,4-D、KT、TDZ和NAA,研究不同植物生长调节剂对太白米愈伤组织诱导的影响.结果显示,4种植物生长调节剂在不同水平上对愈伤组织诱导有显著作用(2,4-D、TDZ,P<0.01;NAA、KT,P<0.05),诱导愈伤组织的最佳激素组合是0.5 mg·L~(-1) 2,4-D+0.5 mg·L~(-1) NAA+0.2 mg·L~(-1) KT+0.1 mg·L~(-1) TDZ,愈伤组织在该培养基上继代培养有不定芽分化,如用相同浓度ZT代替KT则分化的不定芽增多.HPLC分析显示野生小鳞茎、愈伤组织和不定芽中总黄酮含量无显著差异,分别为干重的13.3%、11.4%和11.9%. 相似文献
6.
Anzidei M. Bennici A. Schiff S. Tani C. Mori B. 《Plant Cell, Tissue and Organ Culture》2000,61(1):69-79
Different NAA plus kinetin or BA combinations were tested on Francia Pernod fennel seedlings for callus induction and plant
regeneration. Callogenesis from hypocotyls was obtained in all auxin/cytokinin-containing media. The organogenic response
was observed especially in presence of NAA plus kinetin. The highest frequency of shoot regeneration was found when the auxin
and kinetin were used at a 1:1 ratio. Moreover, a prolonged culture period increased shoot formation. Somatic embryogenesis
was tested on several fennel populations. The results gave evidence of the genotypic importance. Two different protocols were
used for somatic embryo induction. Using the first protocol among the different fennel genotypes tested, only Francia Pernod
showed embryogenic capacity. In this case, from a primary non-embryogenic callus cultured for 12 months in presence of 2,4-D,
an embryogenic secondary callus was produced. When transferred to the medium without 2,4-D (agarized or liquid), this gave
embryogenic plants in high frequency. As far as the second embryogenic method is concerned, secondary embryogenic callus developed
only in the presence of 2,4-D plus kinetin in Francia Pernod genotype. Thereafter, the replacement of those growth regulators
by GA3 into the medium greatly increased the somatic embryo development, especially in `Francia Pernod', but also in `Aboca erbe'
callus, a population with a very poor embryogenic capacity. In Francia Pernod, the primary and secondary (embryogenic) calli
showed different morphological and histological responses, either when the secondary callus was induced by 2,4-D alone or
by 2,4-D plus kinetin. Ontogenetic processes leading to somatic embryo formation are described in this context.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
Plant regeneration from Bulgarian rose callus 总被引:5,自引:0,他引:5
Plant regeneration capacity of Bulgarian rose callus tissue was examined. Adventitious bud formation could be successfully attained, depending on the kinds of mineral salts used in the medium, auxin and cytokinin used. When callus tissues were cultured on the medium without ammonium nitrate and contained indoleacetic acid and benzyladenine, buds were formed in the callus. The number of buds were significantly increased by the simultaneous addition of calcium ionophore. When the cultures were transferred to the medium without cytokinin, roots were formed in the basal part of the buds.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- K
kinetin
- NAA
naphthaleneacetic acid 相似文献
8.
S. C. Debnath D. B. McKenzie K. B. McRae 《Journal of plant biochemistry and biotechnology.》2001,10(1):57-60
Combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA) affected the callus formation from stem, rachis and leaf explants of beach pea. Mature leaf pieces were best for inducing a callus; when cultured on Murashige and Skoog’s (MS) medium supplemented with 4.4μM BA at least 87% of the explants callused regardless of the NAA concentration. Optimal induction was achieved with 5.4 to 10.7μM NAA depending on the explants used. BA at 4.4μM was always more effective than at the lower concentration of 1.1μM. Multiple buds were induced from calli and formed shoots when transferred to MS medium supplemented with 2.7μM NAA + 4.4μM BA. 相似文献
9.
魔芋属植物愈伤组织的诱导和再生植株的研究 总被引:10,自引:0,他引:10
用云南魔芋(Amorphophallus yunnanensis)、白魔芋(A.albus)、花魔芋(A.revieri)和勐海魔芋(A.bannaensis)等四个种的叶、鳞叶、花序、匍匐茎、块茎和茎尖为外植体,诱导产生小植物。可通过三种途径获得魔芋再生植株:(1)由外植体诱导愈伤组织,再分化小植株:在添加0.5—1.0mg/1 NAA和BA或KT、ZT的MS培养基上,高频率地诱导瘤状愈伤组织形成,发现NAA对愈伤组织的诱导是必不可少的,而细胞分裂素与之组合,促进瘤状愈伤组织的形成和发育,在无激素或低浓度激素的培养基上,瘤状愈伤组织分化出小植物;(2)茎尖和鳞叶、块茎切块培养,诱导形成多芽苗,产生再生植株;(3)块茎切块诱导产生微型小块茎,然后分化芽和根,长成小植物。本研究为魔芋的快速繁殖提供了新的手段。 相似文献
10.
Leaf explants of Solanum khasianum regenerated on MS medium containing 2, 4-D (3 mg/l) and kinetin (1 mg/l). Shoots could be induced from these calli on medium containing BAP (3 mg/l) alone. Rhizogenesis of these shoots occurred when transferred to medium containing 2 mg/l NAA. The yield of solasodine — a pharmaceutically important compound, from 4-month-old callus tissue was remarkable at 2 per cent of dry weight. 相似文献
11.
Calli were induced from cotyledon segment of mung bean (Phaseolus radiatus L.) in Miller medium supplemented with NAA 4 mg/l, kinetin 10 mg/L. The callus formation was completely prevented by the addition of actinomycin D 15 μg/mL or cyclo- heximidc 0.5 μg/mL at 0 hour. The inhibitory effect of actinomycin D or cycloheximide was increased with the increment of concentration but decreased when the inhibitory agents were added a few hours later. If actinomycin D or cycloheximide was added at 24 hour culture it inhibits neither the induction of callus formation nor the proliferation. The content of RNA, DNA and protein were determined. RNA in each segment increased obviously in the early stage of callus formation, but DNA and protein increased slightly afterward. It is suggested that a large increase of RNA is the characteristic of dedifferentiation of cotyledon in P. radiatus. In addition, it has also been shown that an actinomycin D or cycloheximide-sensitive process in the early stage of dedifferentiation is crucial for the callus formation. Both RNA and protein synthesis are required for the initiation of dedifferentiation. 相似文献
12.
13.
C. Pugliesi M. Rabaglio F. Cecconi S. Baroncelli 《Plant Cell, Tissue and Organ Culture》1992,28(2):125-128
Different plant explants of Persian buttercup (Ranunculus asiaticus L.) were screened for callus induction and adventitious shoot regeneration on different media to establish totipotent cultures. Murashige & Skoog (MS) medium was used, supplemented with different concentrations of the following growth regulators: kinetin, benzyladenine (BA), naphthaleneacetic acid (NAA) and indoleacetic acid (IAA). Callus was induced and adventitious buds regenerated only from cotyledonary explants after 4–5 weeks. Subculture of the regenerated buds on the same basal medium in presence of gibberellic acid (GA3) and BA produced well-organized shoots. Rooting was obtained by transferring shoots to growth regulator-free MS medium. A high rate of shoot multiplication has been achieved on medium with high concentration of kinetin and long-day photoperiod. Finally the plants were successfully transferred to soil and grown in a greenhouse. 相似文献
14.
Immature inflorescences of kodo millet (Paspalum scrobiculatum L. cv. GPUK-3) were cultured on MS medium. Induction of embryogenic callus and subsequent somatic embryogenesis was possible
on both 2,4-D and Picloram alone or with kinetin from spikelets as well as rachis. Immature inflorescence cultured on medium
supplemented with lower levels of Picloram in combination with kinetin developed organogenic callus with shoot buds. Direct
somatic embryo formation on rachis was observed at higher levels of Picloram in combination with kinetin. Plant regeneration
was observed when calluses were transferred to α-napthaleneacetic acid (NAA) plus 6-benzylaminopurine (BA) supplemented MS
medium. Histological observations provided a clear evidence for both somatic embryogenesis and shoot organogenesis. Profuse
rooting was induced on phenylacetic acid (PAA) supplemented medium. Regenerated plants were successfully transferred to pots
under field conditions where most of the plants survived and set normal seeds. 相似文献
15.
Cui Yuan-fang Gong Zheng Chen Xin-hua Shao Xing-ti Zhu Zhi-qing Chu Chih-ching 《植物学报(英文版)》1986,28(6)
This paper deals with the preliminary results on callus induction and plant regeration from embryo in vitro of Elaeis guineenis. When mature embryos were cultured on dedifferen- tiation medium they proliferated calli during 30-90 days of culture. Among auxins applied, 2,4-D was more important for callus induction however 2,4-D and NAA combination gave bet- ter result. On the contrary, kinetin inhibited callus formation and growth. These experimental results explain that a higher callus induction frequency depends not only on the constituents of the medium used but also on the genotype of donor plants. After transfering the calli onto kinetin-containing media for a peirad, embryoids, which showed typical configuration of zygotic embryo, could be obtained. The embryoids can further develop into whole plants on a shoot induction medium. Some embryoids have subjected to srveral generations of subculture and still retained the ability to embryoid multification and plant regeneration. 相似文献
16.
Leaf, stem, hypocotyl, cotyledon, root, shoot tip and embryo explants of Capsicum annuum L. cv. mathania were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) or kinetin (Kin) alone or in
combination with 3-indoleacetic acid (IAA), 3-indolebutyric acid (IBA), α-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic
acid (2,4-D). BAP (5.0 mgl−1) in the medium was found to be the best growth regulator for shoot bud differentiation. Shoot buds cultured on 5.0 mgl−1 BAP increased in number but did not elongate. For obtaining complete plantlets, shoot buds were placed on a medium with IBA
or NAA (0.1 mgl−1). Histological evidence revealed direct differentiation of buds from cotyledons. Regenerated plants were normal diploids.
Unorganized callus could not be induced to differentiate shoot buds. 相似文献
17.
Jinfeng Lü Rong Chen Muhan Zhang Jaime A. Teixeira da Silva Guohua Ma 《Journal of plant physiology》2013
Camellia nitidissima Chi (Theaceae) is a world-famous economic and ornamental plant with golden-yellow flowers. It has been classified as one of the rarest and most endangered plants in China. Our objective was to induce somatic embryogenesis, shoot organogenesis and plant regeneration for C. nitidissima. Three types of callus (whitish, reddish and yellowish) were induced from immature cotyledons on improved woody plant medium (WPM) with different plant growth regulators (PGRs). Among the callus, whitish callus was induced by 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and reddish and yellowish callus were induced by strongly active cytokinins, thidiazuron (TDZ) or 6-benzylaminopurine (BAP), singly or combined with weakly active auxin, α-naphthaleneacetic acid (NAA). The embryogenic callus could differentiate into somatic embryos, nodular embryogenic structures (large embryo-like structures) or adventitious shoots depending on the PGR used in WPM. BAP was best for adventitious buds and zeatin was best for somatic embryogenesis while kinetin (Kt) was best for the formation of nodular embryogenic structures. The three regeneration pathways often occurred in the same embryogenic callus clumps. Most shoots (80.0%) developed roots in WPM supplemented with 24.6 μM IBA and 0.3 μM NAA while 47.5% of somatic embryos could germinate directly and develop into plantlets on induction medium supplemented with 0.9 μM BAP and 0.1 μM NAA. The nodular embryogenic structures could be sub-cultured and cyclically developed in one of two differentiation pathways: shoot organogenesis or somatic embryogenesis. Plantlets derived from shoot buds rooted and somatic embryos germinated when transplanted into soil in a greenhouse; 66.7% of plantlets from shoot culture and 78.6% of plantlets from somatic embryos survived after 8 weeks’ acclimatization. 相似文献
18.
Protoplasts were isolated from spongy calli in a well growing state. Protoplasts were induced to undergo sustained divisions and to form colonies in the liquid C81V medium supplemented with 2,4-D and kinetin. When protoplast derived colonies were transferred onto agarsolidified medium, the spongy, white calli developed. After being subcultured on N6 medium plus 6BA and IBA, the light-yellow, granular embryogenic calli emerged on the protoplast regenerated callus surface. A large number of plantlets were obtained on MS medium with NAA and IBA via somatic embryogenesis Cytological observation on the donor calli used for protoplast isolation and plantlets regenerated from protoplasts were carried out. Remarkable variation of nucleus morphology and chromosome numbers were observed in donor calli. However, the cytological abnormalities in plantlets regenerated from protoplasts were comparatively less seen. The reason are discussed. 相似文献
19.
20.
The endosperm culture of wheat-rye hybrid was studied in order to explore a new pathway of chromosome engineering. The preliminary results were obtained to show that the endosperm callus formation could be induced from the young endosperm within 7–14 days after crossing on the medium supplemented with 2 ppm 2,4-D, 0.5 ppm kinetin and 3%–8% sucrose. The induction frequency of callus amounts to 35.3%. When the calli were transfered onto an auxin step-down medium containing 0.5 ppm IAA and 1 ppm kinetin, both shoots and roots were formed. 4 endosperm plantlets were obtained. The chromosome number in somatic cells of endosperm plantlets was very unstable. The numbers varied from 6—42, but there is no 49 to be found. The chromosome number with 1—4 times of 7 can be found in higher percentage. 相似文献