In-vitro developmental biology- Embryo (original) (raw)
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Plant regeneration from embryogenic suspension cultures of Musa acuminata cv. Mas (AA
Plant Cell Tissue and Organ Culture, 2003
Embryogenic callus was established using immature male flower of Musa acuminata cv. Mas. After 5-6 months of culture, embryogenic callus was obtained at 21.75%Ϯ11.9 from 750 immature male flower clusters with translucent somatic embryos proliferated from the whitish friable callus. It was observed that flower clusters ranging from 4 to 11 responded to form embryogenic callus and out of which 3-10 somatic embryos were formed per flower cluster. Embryogenic callus were obtained at a percentage of 10.00%Ϯ0.3 on M1 medium initially supplemented with 18 M 2,4-dichlorophenoxyacetic acid (2,4-D) for 3 months and subsequently transferred to the same media with reduced 2,4-D (9 M) for the next 2-3 months. Embryos developed into translucent spheres and slightly torpedo shaped embryos in suspension cultures. Plantlets were obtained on medium M4 supplemented with 0.8 M BA, at an average regeneration rate of 13.00%Ϯ0.58.
Euphytica, 1996
Plant regeneration was achieved in Verbascum speciosum Schard. via organogenesis and somatic embryogenesis by culture of mature embryo explants. Two types of calli, embryogenic and non-embryogenic, were induced from mature embryo explants on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyl adenine (BA) and α-naphthalene acetic acid (NAA). In order to further proliferate the somatic embryoids, the yellow and friable embryogenic calli were transferred on MS medium containing 0.5 mg-1 charchol and 0.1 or 1 mg-1 2,4-dichlorophenoxy acetic acid (2,4-D) or into MS medium containing 60 g-1 sucrose, 50 mgl-1 casein hydrolysate (CH), 0.5 mg-1 kinetin (Kin), 5 mg-1 2,4-D and 0.5 mg-1 charchol. Among the 3 tested media, MS medium containing 0.1 mg-1 2,4-D and 0.5 mg-1 charchol was more effective for proliferation of embryonic calli. Somatic embryos were transferred to hormone free MS medium for maturation and shoot regeneration. In addition, shoots and roots regenerated from non-embryogenic calli in hormone free MS medium or containing NAA and BA. Shoot buds were obtained from non-embryogenic calli and they were transferred to MS medium supplemented with 1 mg-1 BA or Kin for further growth and multiplication. Regenerated plants then were potted and maintained in the greenhouse.
Cymbopogon schoenanthus subsp. proximus has been achieved. The species is rare and confined in its distribution to Africa. A range (0.5 to 8 mg/l) of 2,4-dichlorophenoxyacetic acid (2,4-D) for the induction of embryogenic callus from seed cultures were used. Results show that 1.0 and 4.0 mg/l 2,4-D gave a 90 to 100% frequency of embryogenic callus containing mature embryos within three months of culture. Testing the effect of phosphorus and nitrogen concentrations on somatic embryogenesis from callus cultures showed that high phosphorus and low nitrogen concentrations enhanced embryo induction. Low NH 4 NO 3 enhanced growth and maturation of somatic embryos, while low KNO 3 enhanced germination and shoot production. Suspension cultures were initiated from embryogenic callus on 0.5, 1.0 and 2.0 mg/l 2,4-D, then plated on 3 different combinations of 2,4-D and 6-benzyl adenine (BA). Mature embryos were highest on both 0.5 and 1.0 mg/l 2,4-D (X = 3), while shoot germination was enhanced by using BA in the regeneration media. The effect of high phosphorus concentration in the culture media was significant on both embryo induction and early maturation (X = 41). Lower ammonium nitrate concentrations enhanced growth and maturation of embryos. Both embryo maturation and the germination of shoots after 6 and 8 months of plating were best on low ammonium nitrate (X = 36.8 and 27.6 shoots, respectively). The presented embryogenic system will be of value for the clonal propagation, ex situ conservation and production of bioactive compounds from this threatened plant species.
For understanding the effect of cloned genes and related molecular mechanisms an efficient and reproducible regeneration and stable genetic transformation protocol is necessary. The crucial factors for enhancing successful regeneration are genotypes, explants tissue, combination and concentration of growth regulators, and culture conditions.The present study was undertaken to optimize the concentration of growth regulators for callus induction and subsequent organogenesis in OryzarufipogonL. WRA21.Calli were induced from seeds of WRA21 on MS medium supplemented with 2,4-D (3.0 mg/l), BAP (0.25 mg/l) and proline (0.6 g/l) gelled with gelrite (3.0 g/l), after 10 days of seed culturing. The embryogeniccalli were regenerated under subdued light at 25°C ± 2°C on MS regeneration medium supplemented with BAP (3.0 mg/l), NAA (0.2 mg/l) and gelled with agar (8 g/l) in combination with gelrite (2.0 g/l) and regeneration occurred in 20 days. The regeneration percentage achieved was very high (82.66%). The plantlets were hardened and transferred to soil in pots. The regeneration protocol so developed through embryogeniccalli was highly reproducible. The plants showed normal growth and flowering under greenhouse conditions.
The Malnad region located in the Western Ghats of Karnataka is known for the cultivation of indigenous rain fed land race cultivar of rice. The present study was to investigate the callogenic and caulogenic potentialities of the two indigenous rice cultivar namely Karimundaga and Kanadatumba using dehusked mature embryo explants. For callus and shoot bud differentiation, the explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-D (1–3 mg/L), IAA (1–2 mg/L), Kn (1–4 mg/L) and BAP (1–4 mg/L). The morphogenic potentialities of the two rice cultivar differed in texture of callus. In both the cultivar callogenic frequency was optimized at 1 mg/L 2,4-D concentration, it was 94% in Karimundaga and 58% in Kanadatumba. Supplementation of IAA either alone (1–2 mg/L) or in combination with Kn or BAP at 1 to 4 mg/L concentration of each induces shoot bud differentiation from the calli. In the cultivar Karimundaga caulogenic frequency was highest (10.60±2.55) at 1.0 mg/L IAA and 4.0 mg/L BAP concentration. While in the cultivar Kanadatumba highest number of shoot buds (7.90±2.69) was differentiated at 1.0 mg/L IAA and 4.0 mg/L Kn concentration. The calli derived regenerants were successfully acclimatized in the greenhouse and agro-morphological variations were evaluated. The growth characteristics and yield related parameters exhibited by in vitro plants were lower than the in vivo plants.
Plant Regeneration by Somatic Embryogenesis in Azadirachta indica A.Juss. (Neem
2017
From the earliest period people using plant based medicine for their primary health care. Out of them neem tree (Azadirachta indica A. Juss.) belongs to the family Meliaceae is one of the important medicinal plant using from ancient Vedic period in different ailments and now it has a great demand in commercial purpose due to its active medicinal compounds (e.g. Azatin, Azadirachtin).The conventional method of propagation is not sufficient to overcome the requirements of neem tree. Somatic embryogenesis is a potential tissue culture method which can be utilized in regeneration of neem tree for high rate of multiplication. The present observation revealed that the concentration of BA (0.5mg/l and 1.0mg/l) and 2, 4-D 0.1mg/l in individual MS medium were responded to best callus from the inoculated explants i.e. leaf and stem. The MS medium supplemented with 0.5mg/l BA was showed earlier response to callus formation in both leaf (in 8 days) and stem (in 11 days). Whereas combination of IAA and BA was not resulted good effects. The callus grown on MS medium with 2, 4-D showed white color instead of normal green color. Out of the various growth regulators treatment only BA supplemented MS medium stimulated to Somatic embryos (SEs) and 0.5mg/l concentration responded earlier than 1.0mg/l BA. It was resulted that MS medium supplemented with BA as growth regulator is suitable for germination of SEs of Azadirachta indica and 3 rd subculture of callus is optimum for maximal SEs germination.
International Journal of Science and Research (IJSR), 2015
An in vitro plant regeneration system was developed via direct somatic embryogenesis from different seedling explants of an important medicinal plant Murraya koenigii (L.) Spreng Cotyledons (COT), Hypocotyl (HYP) (10 to 15 mm) and Root (RT) segments (10 to 20 mm) were excised from 60 days old seedling as explants. The somatic embryos induction was achieved on Murashige and Skoog (MS) basal medium augmented with different concentrations of 6-benzyleaminopurine (BAP) 1.33 to 8.40 μM and thidiazuron (TDZ) 1.08 to 9.82 μM. The globular embryos originated from cut ends and entire surface of the root, hypocotyl explants and margins of cotyledons within 30-40 days. The percentage of somatic embryos induction per explant was significantly higher in HYP explants (94.21±5.77%) in the MS basal medium supplemented with 6.20 μM BAP and 8.64 μM TDZ. The highest rate of conversion of torpedo, heart and cotyledonary stages from globular stage was obtained in MS medium supplemented with 8.64 μM TDZ. The matured somatic embryos were transferred to the MS basal medium without Plant Growth Regulators (PGRs). Highest 88% of the matured embryos were germinated on transfer to ½ MS basal medium without PGR, where they grew for a further 3-4 weeks. Out of seventy six hardened plants seventy (92%) plantlets were found healthy under field conditions.
In Vitro Cellular & Developmental Biology - Plant, 2004
In vitro regeneration of plants via somatic embryogenesis through cell suspension culture was achieved in horsegram. Embryogenic calluses were induced on leaf segments on solid Murashige and Skoog (MS) medium with 9.0 mM 2,4dichlorophenoxyacetic acid (2,4-D). Differentiation of somatic embryos occurred when the embryogenic calluses were transferred to liquid MS medium containing 2,4-D. Maximum frequency (33.2%) of somatic embryos was observed on MS medium supplemented with 7.9 mM 2,4-D. Cotyledonary-torpedo-shaped embryos were transferred to liquid MS medium without growth regulators for maturation and germination. About 5% of the embryos germinated into plants, which grew further on solid MS medium. The plants were hardened and established in soil. Effects of various auxins, cytokinins, carbohydrates, amino acids, and other additives on induction and germination of somatic embryos were also studied. A medium supplemented with 7.9 mM 2,4-D, 3.0% sucrose, 40 mg l 21 L-glutamine, and 1.0 mM abscisic acid was effective to achieve a high frequency of somatic embryo induction, maturation, and further development.
Plant Cell, Tissue and Organ Culture, 1992
The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum x A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid. The embryogenic calluses of all three Allium species were similar in appearance. For all accessions tested plants could be regenerated at a high frequency from this compact callus through somatic embryogenesis, when using kinetin supplemented MS medium (regeneration medium). Addition of abscisic acid to the regeneration medium stimulated the formation of both somatic embryos and shoots for a number of varieties. Concerning shoot regeneration from callus cultures, significant differences existed between genotypes of all accessions except one.
Industrial Crops and Products, 2017
Ajowan (Carum copticum L.) is an important and endangered industrial medicinal plant that growing in some parts of Iran. Two efficient protocols, without somaclonal variation induction, were developed for indirect somatic embryogenesis and indirect shoot regeneration of three Iranian ecotypes of ajowan. In the first experiment, higher concentration of auxin than cytokinin (1, 1.5 and 2 mg/L of 2,4-dichlorophenoxyacetic acid along with 0.25, 0.5 and 0.75 mg/L kinetin) was used for callus induction in 5, 10 and 15 days old hypocotyl explants. In the second experiment, higher concentration of cytokinin than auxin (1 mg/L of kinetin along with 0.5 mg/L of 2,4-dichlorophenoxyacetic acid) was used for callus induction in 15-d old hypocotyl explants. The higher frequency of somatic embryos was achieved from 15 days old hypocotyl explants of Ghoom ecotype with 28.33 embryos when induced calli from MS medium supplemented with 1.5 mg/L 2,4-dichlorophenoxyacetic acid in combination with 0.5 mg/L kinetin transferred to free plant growth regulator MS medium. Momentary removing of 2,4dichlorophenoxyacetic acid was successful for somatic embryogenesis in Iranian ecotypes of ajowan that it significantly reduce the time of the culture and thus reduce the risk of somaclonal variation. Maximum number of initiated shoots per explant was related to Shiraz ecotype with average 18.33 shoots per callus in MS medium supplemented with 1.5 mg/L of specify type of cytokinin (3-methoxy [-6-benzylamino-9-tetrahydropyran-2-yl] purine) plus 0.25 mg/L naphathalene acetic acid. The survival rate of rooted plantlets was 60.86% and 58.33% for indirect somatic embryogenesis and indirect shoot regeneration derived plants, respectively. The genetic stability of regenerated plants via indirect somatic embryogenesis and indirect shoot regeneration was proved through flow cytometry analysis.