Indirect shoot regeneration of Iranian purple coneflower (Echinacea purpurea L.) from cotyledon and hypocotyl explants

2011 ◽  
Vol 59 (1) ◽  
pp. 65-72 ◽  
Author(s):  
A. Zebarjadi ◽  
J. Motamedi ◽  
A. Ismaili

In vitro plant regeneration was optimized for Iranian purple coneflower via organogenesis from callus cultures derived from cotyledon and hypocotyl tissues by placing them on MS medium supplemented with different concentrations and combinations of BAP and NAA. The experiment was laid out as a completely randomized design in a factorial arrangement with three replications. The results indicated that the mean callus induction was influenced by explant type, with a significant difference between cotyledon (77.81%) and hypocotyl (65.33%) explants at the 0.01 probability level. In relation with the regeneration rate, no significant differences were observed between the two types of explants. For both cotyledons and hypocotyls the optimum shoot regeneration frequency (31.5% and 32.5%, respectively) and number of shoots per explant (5.2 and 5.3, respectively) were achieved using medium supplemented with 0.4 mg l−1 BAP. Proliferated shoots were elongated in hormone-free MS medium and well-developed shoots were rooted on MS medium, both with and without the addition of 2 mg l−1 IBA. All the plantlets survived acclimatization, producing normal plants under controlled conditions. This study revealed that cotyledon and hypocotyl explants of E. purpurea have relatively good potential for callus induction and shoot formation. Furthermore, a beneficial method has been established for the micropropagation of this valuable medicinal species.

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501
Author(s):  
Sanjog T. Thul ◽  
Arun K. Kukreja

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.


2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.


1970 ◽  
Vol 8 (1) ◽  
pp. 1-6 ◽  
Author(s):  
M Hoque ◽  
KM Nasiruddin ◽  
GKMN Haque ◽  
GC Biswas

The experiment was conducted during May to December 2008 in the Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh to observe the callus induction, regeneration potentiality and to establish a suitable in vitro plantlet regeneration protocol of Corchorus olitorius. MS medium supplemented with different phytohormone concentrations and combinations were used to observe the callus induction, shoot regeneration and root formation ability of the cotyledon with attached petiole derived explant of three genotypes viz. O-9897, O-72 and OM-1. The highest callus induction (92.85%) was observed in O-9897 followed by O-72 (82.14%) in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype O-9897 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (83.33%) followed by O-72 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on halfstrength of MS media supplemented with 0.6 mg/L IBA in genotype O-9897 (45.00%). The in vitro regenerated plantlets from the genotypes O-9897 could be established in the field. Therefore, the genotypes O-9897 of C. olitorius in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA could be used for callus induction and shoot regeneration. Keywords: Regeneration; Phytohormone; Corchorus olitorius DOI: 10.3329/jbau.v8i1.6390J. Bangladesh Agril. Univ. 8(1): 1-6, 2010


2021 ◽  
Vol 12 ◽  
Author(s):  
Denis Okello ◽  
Sungyu Yang ◽  
Richard Komakech ◽  
Yuseong Chung ◽  
Endang Rahmat ◽  
...  

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.


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


2012 ◽  
Vol 40 (1) ◽  
pp. 107 ◽  
Author(s):  
Cuiqin YANG ◽  
Yaoguo QIN ◽  
Xin SUN ◽  
Shu YUAN ◽  
Honghui LIN

An efficient protocol was established for Sedum spectabile Boreau propagation. Various leaf parts were used as explants to regenerate plantlets, the stem segments of which were cultured for shoot proliferation and plantlet multiplication. The results showed that the leaf base was the optimal explant, as compared to both the middle and the top of leaves, for shoot formation. The highest shoot induction of 88.9% was observed on MS medium supplemented with 0.6 mg/l TDZ and 0.1 mg/l NAA. Hyperhydric leaves obtained in primary culture developed first into abnormal somatic embryos 10 days after subculture, and then into hyperhydric plantlets after an additional 10 days. The hyperhydric plantlets reversed to normal plantlets when plant growth regulators were removed from culture medium. Further, stem segments from reversed plantlets were used for shoot regeneration and root induction. Optimal shoot regeneration was obtained in MS medium containing 0.6 mg/l TDZ with 0.1 mg/l NAA. Root induction and root mean number were all higher on auxin-free medium than on medium containing auxins.


2018 ◽  
Vol 17 (1) ◽  
pp. 37-46
Author(s):  
Attachai TRUNJARUEN ◽  
Sayam RASO ◽  
Pitakpong MANEERATTANARUNGROJ ◽  
Worasitikulya TARATIMA

Current study determined the effects of different cultivation media and some factors on in vitro callus induction and shoot regeneration of Pakaumpuel rice (Oryza sativa L.), the Thai rice landrace. Pakaumpuel’s dehusked seeds were surface sterilized by 20% Sodium hypoclorite for 20 mins before washed and cultured on Murashige and Skoog (MS) medium in various concentrations of BAP (0, 0.1, 0.5 and 1 mg/l) and 2,4-D (0, 1, 2 and 3 mg/l) and cultivated  in off -light or dark cycle condition for 15 days. Results showed that MS medium with 2 mg/l 2,4-D incubated in light condition provided the highest callus induction percentage (80 %). For shoot regeneration, calli were desiccated by using silica gel for 0, 30, 90 and 180 minutes prior to culture on MS medium with 1 mg/l NAA and various concentrations of BAP (0, 1, 2, 3 and 4 mg/l) for 30 days. The results revealed that desiccation periods had no effect on regeneration percentage and number of shoot per callus, but had effects on shoot length and number of leaves per shoot. Therefore, MS medium with 1 mg/l NAA and 3 mg/l BAP induced the highest regeneration percentage (53.33 %) and number of shoot per callus (4.40 shoots), while the longest shoot (12.82 cm) and the highest number of leaves per shoot (3.00 leaves) were found on the culture on this medium after desiccation for 90 minutes. This discovery can be applied for micropropagation of other Thai rice landraces.


Author(s):  
Vi Thi Tuong Nguyen ◽  
Trinh Le Diem Ho ◽  
Kim Thi A Phan

Codonopsis javania (Blume) Hook.f. et Thomson a traditional medicine plant and now an endangered species in Vietnam is grown for roots. The research was carried out to establish the plant propagation for the purpose of concerving and exploting this endangered medicinal herbs. In vitro shoot tip explants (1 – 1.5 cm) were induced to form callus on MS medium containing NAA (0.5 – 2 mg /L) with TDZ 0.1 mg/L. After four weeks of culture, in the MS medium combine with NAA 1 mg/L and TDZ 0.1 mg/L the explant induced compact callus (green, solid) wsa achieved 85.33%. The callus induction to form shoots on medium MS containing BA (0.5 – 2.0 mg/L) with NAA 0.2 mg/L. After 4 weeks of culture, shoot formation was higher in the MS medium containing BA 1.0 mg /L and NAA 0.2 mg/L and achieved of 82.67 % with 9.92 shoots/explant. The best shoot proliferation (2 – 3 cm) was excised and transferred to a medium shoot multiplication with the same composition as the shoot induction medium in which NAA 0.2 mg/L was replaced by NAA 0.5 mg/L. When compared the shoot multiplication between the two mediums at the same BA concentration (2 mg/L), all shoots increased and reached 5.87 times after 60 days cultured. On rooting MS medium with IBA 1 mg/L, 88.67 % in vitro rooting was observed with the average root yield of 4.33 roots/shoot and the length of 8.27 cm. Root length and their yield quality were highly improved when using of coconut fiber (30 %) and earthworms compost (70 %) (v/v) in the transfer medium after acclimatisation stages.


2017 ◽  
Vol 15 (10) ◽  
pp. 701-710
Author(s):  
Piyaporn SAENSOUK ◽  
Surapon SAENSOUK ◽  
Phattaraporn PIMMUEN

An efficient and rapid protocol for the micropropagation of Globba schomburgkii Hook. f. via bulbil explants was investigated. The long divided and undivided bubils of G. schomburgkii Hook. f. were cultured on MS medium (Murashige and Skoog) that had either 3 mg/l benzyladenine (BA) or 0.5 mg/l naphthaleneacetic acid (NAA) added for 8 weeks. The results indicated that the long divided bulbils of G. schomburgkii Hook. f. showed a greater amount of plant regeneration than the undivided bulbils. Callus induction, as well as shoot and root formation, were observed when culturing microshoots of 1 cm in length on media (MS) that had Thidiazuron (TDZ) or NAA plus BA added at a range of concentrations for 8 weeks. The highest percentage of callus induction was 40 % when culturing the microshoots on MS medium supplemented with NAA and BA. The best result for shoot formation was achieved when culturing the microshoots on MS medium with TDZ added. The highest number of roots was obtained when culturing the microshoots on MS medium with NAA and BA added. The in vitro-derived plantlets of G. schomburgkii Hook. f. were transplanted to pots containing different types of potting mixture in a greenhouse. The survival rates were 80 % when G. schomburgkii Hook. f. was transplanted to sand.


2017 ◽  
Vol 8 (2) ◽  
pp. 20 ◽  
Author(s):  
Supria Saha ◽  
Zohorul Islam ◽  
Sadequl Islam ◽  
Mirza Fida Hassan ◽  
Md. Shahadat Hossain ◽  
...  

A suitable plant regeneration system has been established using 3-4 weeks old calli derived from immature and mature seeds of four wheat varieties viz. Pavon 76, Akbar, Barkat, and Kanchan. As plant growth regulators, various auxins (2,4-D, BAP and IAA) either single or in combination were used in MS medium. The variety Pavon 76 showed maximum (72.25%) callus induction and Akbar exhibited the lowest (37.78%) from calli derived from immature seeds. Hormonal effects on callus induction were evaluated and significant results were found in case of genotypes at P <0.01. Out of four genotypes, the highest frequency of plant regeneration was recorded in Pavon 76 (67.00%) and lowest in Kanchan (43.10%) when 1.5 mg/l BAP and 0.5 mg/l IAA was added in the medium. It was observed that Pavon 76 produced highest number of green plants than others. For mature seeds all of the mentioned genotypes showed significant difference with maximum frequency of callusing in Pavon 76 (69.57%) in MS + 2.5 mg/l 2,4-D followed by Kanchan (60.84%), Barkat (52.73%), and Akbar (47.19%). For plant regeneration, Pavon 76 also showed best performance (64.36%) in MS + 2.0 BAP + 1.0 mg/l IAA, using calli derived from mature seeds. The other genotypes Barkat, Kanchan and Akbar exhibited 59.44, 52.71 and 52.32% regeneration in the same medium respectively. Here, the lowest regeneration (40.63%) was found in Akbar. In this case, it was aimed to establish a suitable protocol for in vitro callus induction and regeneration for advance biotechnological research on wheat in Bangladesh.


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