scholarly journals MICROPROPAGATION OF PHALAENOPSIS SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

2018 ◽  
Vol 6 ◽  
pp. 1185-1191
Author(s):  
Minh Van Tran

Phalaenopsis spp. was regularly produced through micropropagation by protocorm like bodies (PLBs); micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation. The method involved using protocorm like bodies as planting materials. PLBs were cut into slices and placed on the medium for callus initiation. The callus was initiated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l) or 2,4D (1 mg/l) and was proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (0.5, 1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the MS medium supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Embryonic cell suspensions were plated and regenerated on the medium: 1/2MS supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Micropropagation of Phalaenopsis sp. via the embryogenesis technique was set up to produce 5,800 plantlets per one liter of somatic embryogenesis suspension.

2018 ◽  
Vol 6 ◽  
pp. 1178-1184
Author(s):  
Minh Van Tran

Cymbidium spp. was regularly produced through micropropagation by protocorm like bodies (PLBs) and multiple shoots; micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation to resolve the above difficulties. The method was to use PLBs as planting materials to produce somatic callus cell and embryos. Results were followed as: PLBs were cut into slices and placed on the medium for callus initiated and used as materials for embryo formation study. A fresh weight of callus were used for the experiment of about 100 mg. The medium for the initiation of embryonic callus was composed of: MS + BA (0.1 mg/l) + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1mg/l) or 2.4D (1 mg/l) and was proliferated on the medium MS + pepton (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) or 2.4D (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium consisting of MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) + BA (0.1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with BA (1 mg/l). Embryonic cell suspensions were plating and regeneration on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (0.1 mg/l) + BA (1 mg/l). Micropropagation of Cymbidium spp. via embryogenesis technique was set up to produce 3,800 plantlets per one liter of somatic embryogenesis suspension.


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Nisar Ahmad Zahid ◽  
Hawa Z.E. Jaafar ◽  
Mansor Hakiman

Ginger (Zingiber officinale Roscoe) var. Bentong is a monocotyledon plant that belongs to the Zingiberaceae family. Bentong ginger is the most popular cultivar of ginger in Malaysia, which is conventionally propagated by its rhizome. As its rhizomes are the economic part of the plant, the allocation of a large amount of rhizomes as planting materials increases agricultural input cost. Simultaneously, the rhizomes’ availability as planting materials is restricted due to the high demand for fresh rhizomes in the market. Moreover, ginger propagation using its rhizome is accompanied by several types of soil-borne diseases. Plant tissue culture techniques have been applied to produce disease-free planting materials of ginger to overcome these problems. Hence, the in vitro-induced microrhizomes are considered as alternative disease-free planting materials for ginger cultivation. On the other hand, Bentong ginger has not been studied for its microrhizome induction. Therefore, this study was conducted to optimize sucrose and plant growth regulators (PGRs) for its microrhizome induction. Microrhizomes were successfully induced in Murashige and Skoog (MS) medium supplemented with a high sucrose concentration (>45 g L−1). In addition, zeatin at 5–10 µM was found more effective for microrhizome induction than 6-benzylaminopurine (BAP) at a similar concentration. The addition of 7.5 µM 1-naphthaleneacetic acid (NAA) further enhanced microrhizome formation and reduced sucrose’s required dose that needs to be supplied for efficient microrhizome formation. MS medium supplemented with 60 g L−1 sucrose, 10 µM zeatin and 7.5 µM NAA was the optimum combination for the microrhizome induction of Bentong ginger. The in vitro-induced microrhizomes sprouted indoors in moist sand and all the sprouted microrhizomes were successfully established in field conditions. In conclusion, in vitro microrhizomes can be used as disease-free planting materials for the commercial cultivation of Bentong ginger.


2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Efah FITRAMALA ◽  
Eva KHAERUNNISA ◽  
Nina Ratna Djuita Ratna DJUITA ◽  
Hadi SUNARSO ◽  
Diah RATNADEWI

AbstractBanana (Musa paradisiaca L) cv. Kepok Merah has a high commercial value as it is used in food industries such as banana chip. Besides, Kepok Merah contains high B-complex vitamins that serve in energy metabolism and are important in the development of infant brain. The establishment of industrial plantations of this plant has been restricted by the lack of planting materials. This research aimed at ameliorating the capacity of plantlets multiplication up to rooting of this banana in a rapid way through in vitro multiplication techniques. Murashige and Skoog (MS) and Woody Plant (WP) were used as the basic media. For the initiation stage, the media was fortified with 0.2 mg/L IAA and two levels of BA at 3 and 5 mg/L.  For shoot multiplication, the concentrations of IAA as well as BA were increased. For rooting, 1 mg/L NAA or IBA was applied. The observations demonstrated that for shoots initiation, both basic media performed good results when enriched with 0.2 mg/L IAA and 5 mg/L BA. The highest rate of shoots multiplication at 6 – 17 shoots per explant, was obtained on MS medium added with 0.5 mg/L IAA and 5 mg/L BA.  NAA at 1 mg/L in MS medium produced more rooted plantlets, 3 – 16 roots per plantlet, than those of other treatments. Keywords: Musa paradisiaca cv. Kepok Merah, in vitro micropropagation, scalps.AbstrakPisang (Musa paradisiaca L.) kultivar Kepok Merah memiliki nilai komersial yang cukup tinggi yaitu sebagai bahan dalam industri pembuatan keripik pisang. Selain itu, pisang Kepok Merah memiliki kandungan vitamin B kompleks cukup tinggi untuk membantu produksi energi dan pembentukan sel-sel otak pada bayi. Pertanaman pisang ini dalam skala industri terkendala oleh kurangnya ketersediaan sumber benih. Teknik kultur jaringan diharapkan dapat menghasilkan benih secara massal dalam waktu yang relatif singkat. Tujuan dari penelitian ini adalah meningkatkan keberhasilan multiplikasi tunas in vitro hingga pengakaran tanaman pisang Kepok Merah secara cepat. Pada tahap inisiasi tunas digunakan media dasar Murashige and Skoog (MS) dan media Woody Plant (WP); ke dalam media dasar tersebut ditambahkan IAA 0,2 mg/Ldan 2 taraf BA yaitu 3 dan 5 mg/L. Multiplikasi tunas dilakukan pada media dasar yang sama namun dengan taraf konsentrasi IAA serta BA yang ditingkatkan. Tahap perakaran menggunakan media dasar MS dan WP dengan auksin NAA 1 mg/L atau IBA 1 mg/L. Hasil penelitian menunjukkan bahwa untuk inisiasi tunas, media MS dan WP yang diperkaya dengan IAA 0,2 mg/L dan BA 5 mg/L   sama baiknya. Untuk multiplikasi tunas, media MS dengan IAA 0.5 mg/L   yang dikombinasikan dengan BA 5 mg/L   memberikan jumlah tunas paling banyak, yaitu 6 – 17 tunas per eksplan, dan pertumbuhannyapun lebih baik. Pemberian NAA 1 mg/L   pada media MS dapat memberikan lebih banyak tunas yang berakar, dengan jumlah akar 3 – 16 per planlet.  Kata kunci: Musa paradisiaca cv. Kepok Merah, mikropropagasi in vitro, nodul meristematik


BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Elizabeth Oliver ◽  
João Pedro Alves-Lopes ◽  
Femke Harteveld ◽  
Rod T. Mitchell ◽  
Elisabet Åkesson ◽  
...  

Abstract Background Advances in three-dimensional culture technologies have led to progression in systems used to model the gonadal microenvironment in vitro. Despite demonstrating basic functionality, tissue organisation is often limited. We have previously detailed a three-dimensional culture model termed the three-layer gradient system to generate rat testicular organoids in vitro. Here we extend the model to human first-trimester embryonic gonadal tissue. Results Testicular cell suspensions reorganised into testis-like organoids with distinct seminiferous-like cords situated within an interstitial environment after 7 days. In contrast, tissue reorganisation failed to occur when mesonephros, which promotes testicular development in vivo, was included in the tissue digest. Organoids generated from dissociated female gonad cell suspensions formed loosely organised cords after 7 days. In addition to displaying testis-specific architecture, testis-like organoids demonstrated evidence of somatic cell differentiation. Within the 3-LGS, we observed the onset of AMH expression in the cytoplasm of SOX9-positive Sertoli cells within reorganised testicular cords. Leydig cell differentiation and onset of steroidogenic capacity was also revealed in the 3-LGS through the expression of key steroidogenic enzymes StAR and CYP17A1 within the interstitial compartment. While the 3-LGS generates a somatic cell environment capable of supporting germ cell survival in ovarian organoids germ cell loss was observed in testicular organoids. Conclusion The 3-LGS can be used to generate organised whole gonadal organoids within 7 days. The 3-LGS brings a new opportunity to explore gonadal organogenesis and contributes to the development of more complex in vitro models in the field of developmental and regenerative medicine.


2010 ◽  
Vol 2 (2) ◽  
pp. 60-63 ◽  
Author(s):  
Muhammad AASIM

Cowpea (Vigna unguiculata L.) is an economically important grain legume crop and is an important source of dietary protein in many of the developing countries. The present study reports the effect of pulse treatment duration, concentration of NAA and presence of NAA in the culture medium on shoot regeneration from plumular leaf explant of Turkish cowpea cv. ‘Akkiz’ and ‘Karagoz’. Pulse treatment of mature embryos with 20 mg l-1 NAA for 1 and 3 weeks followed by culturing of plumular leaf explant on MS medium containing 0.25, 0.50 and 1.0 BAP with 1.0, 2.0 and 4.0 mg l-1 NAA promoted somatic embryogenesis in both cultivars. Longer duration of pulse treatment was deleterious resulting in browning and consequently death of the embryos on explants. Pulse treatment with 20 mg l-1 NAA for one week was less deleterious and developed two plantlets after the explants were transferred to MS0 medium after 6 weeks through somatic embryogenesis in cv. ‘Akkiz’. Pulse treatment with 10 mg l-1 NAA for 1 week showed 33.33-50.00% and 25.00-50.00% shoot regeneration frequency in cv. ‘Akkiz’ and ‘Karagoz’ respectively on MS medium containing 0.25-1.00 mg l-1 BAP. Maximum number of 2.50 shoots each per explant were recorded in cv. ‘Akkiz’ and ‘Karagoz’ on MS medium containing 1.00 and 0.50 mg l-1 BAP respectively. Contrarily, maximum shoot length of 8.98 cm of cv. ‘Akkiz’ and 9.42 cm of cv. ‘Karagoz’ was recorded on MS medium containing 0.50 mg l-1 BAP and 1.00 mg l-1 BAP respectively. Regenerated shoots were rooted on MS medium containing 0.5 mg l-1 IBA and and acclimatized in growth room at room temperature where they produced viable seeds.


Author(s):  
N Yogananth ◽  
R Bhakyaraj ◽  
A Chanthuru ◽  
S Parvathi ◽  
S Palanivel

An efficient protocol was devised for rapid callus induction of Solanum nigrum Linn. from young leaves. MS medium supplemented with different concentrations IAA (1-3 mg/l) with BAP (0.5 mg/l) and NAA (1-3 mg/l) with BAP (0.5 mg/l) for callus initiation. The growth of the calli derived from leaves increased with time of incubation and remained almost constant after 30 days. For solasodine estimation, the field grown plant part of young leaves and in vitro callus (0.5 g each) were weighed and extracted thrice with methanol and subjected to HPLC. The solasodine content of field grown leaves extracts was 0.0798 mg g-1 whereas the solasodine content in the in vitro callus extracts were 0.142 mg g-1 in 2.5 mgL-1 IAA + 0.5 mgL-1 BAP, followed by 0.1162 mg g-1 in 2 mgL-1 NAA + 0.5 mgL-1 BAP. Key words: Callus induction; Solasodine; Solanum nigrum; medicinal plant DOI: 10.3126/kuset.v5i1.2850 Kathmandu University Journal of Science, Engineering and Technology Vol.5, No.1, January 2009, pp 99-103


1970 ◽  
Vol 8 (2) ◽  
pp. 227-232 ◽  
Author(s):  
Sonia B Shahid ◽  
SZ Khan ◽  
L Hassan

The experiment was conducted to optimize a suitable protocol for in vitro regeneration in cole crops. Callus initiation was excellent in the variety Early Tropical. Highest percentage of callus proliferation was observed in Early Tropical (75.0%) followed by Tangail Special Pauslali (55.0%) and the lowest in Tara (40.0% ). Maximum callus proliferation (68.5%) was observed in MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. Callus proliferation was lowest (40.0%) in MS + 2.5 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. MS medium supplemented with 3.0 mgL-1 BAP + 1.0 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 was the best for shoot initiation & plantlet regeneration. The highest number of shoots per vial was 7.20 and the lowest number of shoots per vial was 4.40. Among the concentration MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 showed the highest performance of shoots per vial. The variety Tangail Special Pauslali was the best for root initiation. Keywords: Brassica; Cole crops; Callus; In vitro; Regeneration DOI: 10.3329/jbau.v8i2.7930 J. Bangladesh Agril. Univ. 8(2): 227-232, 2010


2015 ◽  
Vol 7 (2) ◽  
pp. 47-52 ◽  
Author(s):  
S Mojumder ◽  
MD Hossain ◽  
MS Haque ◽  
KM Nasiruddin

The experiment was conducted to develop an efficient protocol for in vitro regeneration of mungbean (Vignaradiata) on the aspect of regeneration potentiality of two mungbean varieties (BINA mung 5 and BINA mung 7) as influenced by different combinations of growth regulators supplemented with MS medium. Cotyledon explant of both varieties was used for the present study. Data were collected for various characters of callus initiation, shoot regeneration and root proliferation. Initiation of callus (%) and required days for its initiation and weight of callus were influenced significantly due to the effect of varieties where BINA mung 5 produced more callus induction (40.36%) at minimum requiring time (18.27 days) including heavier sizes of callus (1.54 g) than BINA mung 7 when BINA mung 5 further recorded the longest root (2.92 cm) compare to BINA mung 7. Effect of treatments of the present study were significantly influenced the whole characters regarding callus culture, shoot regeneration and root proliferation. The highest percentage of callus (88.44%) within minimum time (12.53 days) including larger sizes callus (3.521 g) were produced in 1.0 mg L–1 BAP + 2.5 mg L–1 NAA among the treatments while the highest percentage of regenerated shoot (83.44%) at minimum requiring time (17.59 days) and more shoots (7.69 callus–1) were obtained in 1.0 mg L–1 BAP + 2.0 mg L– 1 NAA. Root induction (82.50%), number of roots plantlet–1 (8.469) with minimum requiring time for initiation (14.13 days) and root length (5.250 cm) were the highest in 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP. Incase of interaction, percentage of callus initiation (89.38 %) was the highest in BINA mung 5 treated by 1.0 mg L–1 BAP + 2.5 mg L–1 NAA at requiring minimum time (12.38 days) while same treatment produced the larger callus (3.581 g) among the interactions. The highest percentage (84.38%) and number (7.813 callus–1) of shoot with minimum requiring time (17.50 days) were found from BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. Similarly, the longest shoot (5.58 cm) was produced from the BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. However, root induction (%), roots plantlet–1, days required for root initiation and root length were statistically similar among the whole interaction treatments due to non significant variation. This result mentioned that the variety BINA mung 5 was better than BINA mung 7 for callus induction, shoot regeneration and root initiation while 1.0 mg L–1 BAP + 2.5 mg L–1 NAA, 1.0 mg L–1 BAP + 2.0 mg L–1 NAA and 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP supplemented with MS medium were the best combinations for better callusing, higher ability of shoot regeneration and root proliferation.DOI: http://dx.doi.org/10.3329/jesnr.v7i2.22203 J. Environ. Sci. & Natural Resources, 7(2): 47-52 2014


2019 ◽  
Vol 1 (4) ◽  
pp. 261-269
Author(s):  
Thuy Linh Thi Nguyen ◽  
Ngoc Thi Pham ◽  
Thao Thi Ninh ◽  
Phuong Thao Thi Nguyen

This study was conducted to establish the procedure for in vitro propagation of Japanese cherry (Prunus sp.) to produce large quantity of plantlets and initial planting materials for climate adaptation research of this plant in Hanoi. Single nodal stems were used as the primary explants and initially produced shoots on MS medium supplemented with 1 mg L-1 BA. The highest shoot multiplication rate (9.57 times) was obtained on MS medium containing 1 mg L-1 BA and 0.25 mg L-1 a-NAA after 8 weeks of culture. 100% of the shoots produced roots with a mean of 10.10 roots per plant within 4 weeks on ½ MSM medium with 4 mg L-1 IBA. The survival rate of in vitro derived plantlets after a 6 to 7-week-period of rooting during acclimatization using a soil: coco peat: smoked rice husks (2:2:1, v/v/v) substrate was 100% and acclimatized plantlets showed good growth and development. This is the first report on a practical and efficient in vitro multiplication protocol for Japanese cherry in Vietnam, starting from shoot initiation to establishment of plants under greenhouse conditions.


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