In Vitro Shoot Regeneration of NAA-Pulse Treated Plumular Leaf Explants of Cowpea

2010 ◽  
Vol 2 (2) ◽  
pp. 60-63 ◽  
Author(s):  
Muhammad AASIM
Keyword(s):  
Somatic Embryogenesis ◽  
Shoot Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Grain Legume ◽  
Ms Medium ◽  
Pulse Treatment ◽  
Mature Embryos ◽  
Legume Crop

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.

scholarly journals In vitro callus induction of gerbera from leaf explant

2020 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Sadia Afrin Jui ◽  
Md. Mijanur Rahman Rajib ◽  
M. Mofazzal Hossain ◽  
Sharmila Rani Mallik ◽  
Iffat Jahan Nur ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Callus Induction ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Indole 3 Acetic Acid

The experiment was designed to evaluate the effect of growth regulators on leaf explant of Gerbera for callus induction. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA) were used to initiate cultures. These were added to Murashige and Skoog medium in different combinations and concentrations. Leaf explants cultured on MS medium supplemented with BAP+ 2, 4-D+ IAA in T4 treatment & BAP+ 2,4-D in T5 treatment showed the best results for callus induction. On the other hand callus was induced early in the combination of BA+ 2,4-D + IAA hormone in T5, T9 & T8 treatment respectively. The rate of callus induction was very low in BA + NAA combinations but it was much earlier.   

scholarly journals Efficient Regeneration of Tobacco (Nicotiana tabacum L.) Plantlets from Cotyledon, Hypocotyl and Leaf Explants: An Excellent Model Plant for Gene Function Analysis

2020 ◽  
pp. 1-9
Author(s):  
Md. Shoyeb ◽  
Kanis Fatema ◽  
Md. Abdur Rauf Sarkar ◽  
Atikur Rahman ◽  
Shaikh Mizanur Rahman
Keyword(s):  
Gene Function ◽  
Callus Induction ◽  
Function Analysis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Regeneration Ability ◽  
Ms Medium ◽  
Gene Function Analysis

Tobacco has been widely used as a model plant for stable and non-stable gene function analysis. Successful Agrobacterium-mediated transformation mainly depends on in vitro regeneration of tobacco plant. However, a reliable and standard regeneration protocol of tobacco using multiple explants is limited. In this study, we established a reliable and reproducible regeneration protocol of tobacco using three different explants i.e. cotyledon, hypocotyl and leaf. Preliminary, surface sterilized tobacco seeds were germinated on growth regulator free MS medium. Thereafter, in vitro germinated explants were inoculated into Murashige and Skoog [1] media supplemented with different combination and types of growth regulators for callus induction and subsequent regeneration of plantlets. It was revealed that, regeneration ability of explants is greatly influenced by type and nature of the explant. Among the three explants, higher callus induction (95%) was obtained in MS medium supplemented with 2.0 mg l-1 kinetin + 2.0 mg l-1 IAA from leaf explant. Also, leaf explant exhibited much higher regeneration ability (95%) than hypocotyl (60%) and cotyledon (45%) explants. Significantly highest number of shoots (8.0) were regenerated from leaf explants cultured on MS medium supplemented with 3.0 mg l-1 Kinetin+1.0 mg l-1 IAA compared to the other hormone combinations. Regenerated mature shoots were showed normal root after transferred onto ½ MS medium containing 0.3 mg l-1 IBA. This study will provide valuable information related to in vitro regeneration of tobacco plantlets using cotyledon, hypocotyl and leaf explants and will be used as a standard protocol for Agrobacterium-mediated transformation for gene function analysis.

scholarly journals Explants selection for in vitro propagation of Pachyrhizus erosus L.

2021 ◽  
Vol 13 (1) ◽  
pp. 10844
Author(s):  
Idowu A. OBISESAN ◽  
Ayobola M. A. SAKPERE ◽  
Bamidele J. AMUJOYEGBE ◽  
Michael S. AKINROPO
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Propagation ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stem Explants ◽  
Ms Medium ◽  
Friable Callus ◽  
Pachyrhizus Erosus ◽  
Regeneration Response

Pachyrhizus erosus tuber is rich in protein asides its agronomical value as a legume, but the seeds by which it is propagated have very low viability. This study established sterilization protocol and effect of various concentrations of auxins and cytokinins on callus production and shoot regeneration from explants of P. erosus. Explants and seeds were sterilized using sodiumhypochlorite (NaClO) solution (5, 10 and 15% v/v) for 5 and 10 mins. Nodal, stem and leaf explants from in vitro germinated P. erosus and tuber from field grown plant were sterilized and cultured on Murashige and Skoog (MS) medium (control) and MS combined with different concentrations of auxins (NAA and 2, 4-D) and cytokinin (BA and Kinetin) and the cultured explants were monitored in terms of degree of callus formation, morphology and colour of callus and also for shoot induction. The results showed that seeds of P. erosus sterilized with 10% NaClO solution for 10 mins and germinated in vitro is the best way of getting sterile nodal, stem and leaf explants for the in vitro propagation of the plant, while tuber explants could be sterilized with 15% NaClO for 10 minutes. Nodal explants inoculated in MS medium supplemented with 1.0 mg/L BA gave the highest shoot regeneration response, while stem explants inoculated on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L NAA also gave the highest amount of friable callus. The study concluded that in vitro germinated seeds were the best way of getting explant for P. erosus.

scholarly journals High frequency plant regeneration of Chrysopogon zizanioides via organogenesis and somatic embryogenesis-an underutilized pharmaceutically valuable biofuel plant

2021 ◽  
Author(s):  
M. Merlin Monisha ◽  
M. Prakash ◽  
K.R. Saravanan ◽  
Anandan R
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Ms Medium ◽  
Natural Habitats ◽  
Root Explants ◽  
Chrysopogon Zizanioides ◽  
Wide Range

Abstract Vetiver (Chrysopogon zizanioides) is an essential oil-producing plant that has tremendous application in cosmetics, perfumery, and herbal medicine. Natural sterility and indiscriminate harvests lead to the risk of extinction of plant species in natural habitats. Therefore, a protocol for regeneration systems via organogenesis and somatic embryogenesis using node, leaf, and root explants has been standardized. The highest shoot regeneration frequency (72.2%) through organogenesis was attained from node explants on MS (Murashige & Skoog) medium comprising 2.0 mg L-1 BAP (“6-benzylaminopurine”). Concurrently, leaf explants cultivated on MS medium augmented by 1.0 mg L-1, 2, 4-D (“2, 4-dichlorophenoxyacetic acid”) formed the optimal frequency (75.35%) of white friable compact (WFC) callus. However, the root explant was less responsive for WFC callus induction. Organogenic WFC callus cultivated on MS medium fortified by kinetin (1.0 mg L-1) as well as BAP (1.0 mg L-1) revealed the highest shoot regeneration efficiency (75.49%) with 48 shoots per callus. Adventitious shoots obtained from node and WFC callus of both leaf and root explants cultivated on MS medium increased by NAA (2.0 mg L-1 showed the optimal rooting of 76.97%. Concomitantly, an elevated frequency of somatic embryogenesis (52.50%) was recorded from leaf explants on MS medium using BAP (0.5 mg L-1) & 2, 4-D (1.0 mg L-1). Leaf explants were superior to node and root explants for somatic embryo initiation. The cotyledonary embryos were efficiently germinated into complete plantlets on a hormone-free MS medium. The plantlets gathered from organogenesis & somatic embryo genesis was effectively acclimatized into phenomenally similar plants. This technique may be applicable for wide-range propagation, genetic engineering, and the formation of bioactive compounds.

scholarly journals Effect of Plant Growth Regulators on Somatic Embryogenesis and Plantlet Development of Turkey Berry (Solanum torvum SW)

2021 ◽  
pp. 1-8
Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Solanum Torvum ◽  
Plantlet Formation ◽  
Regenerated Plants

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.

scholarly journals Efficient in vitro propagation from preconditioned embryonic axes of Turkish cowpea (Vigna unguiculata L.) cultivar Akkiz

2010 ◽  
Vol 62 (4) ◽  
pp. 1047-1052 ◽  
Author(s):  
Muhammad Aasim ◽  
Mahmood Khawar ◽  
Sebahattin Özcan
Keyword(s):  
In Vitro Propagation ◽  
Room Temperature ◽  
Shoot Length ◽  
Grain Legume ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Embryonic Axes ◽  
Legume Crop ◽  
Soil Mix

Cowpea is an important grain legume crop. The study reports an efficient in vitro multiplication and shoots regeneration protocol from preconditioned embryonic axes of the Turkish cowpea cultivar Akkiz. The embryonic axes were preconditioned with 10 mg/l BA on agar solidified MS medium for 5 days. Thereafter they were cultured on MS medium containing 0.25, 0.50, 0.75 and 1.00 mg/l BA with or without 0.10 mg/l NAA. Mean frequency (%) of shoot regeneration, number of shoots per explant and shoot length decreased with each increase in BA concentration used singly. However, a positive increase was recorded in all parameters in the presence of 0.10 mg/l NAA in the regeneration medium. A maximum mean number of 10.33 shoots per explant was recorded on an MS medium containing 1.00 mg/l BA -0.1 mg/l NAA. Regenerated shoots were rooted on an MS medium containing 0.50 mg/l IBA. Rooted plants were acclimatized at room temperature in soil mix contained in pots where they were subjected to an intermittent mistwater spray for 24 h that maintained 90% relative humidity during the first few days which was gradually reduced to 40% for 10 days. All plants flowered and set seeds in a greenhouse after 3 months.

scholarly journals (291) Plant Regeneration through Direct Somatic Embryogenesis in Homalomena `Emerald Gem'

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Direct Somatic Embryogenesis ◽  
Induction Medium ◽  
Ms Medium

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.

scholarly journals Somatic Embryogenesis and Plant Regeneration in Viola canescens Wall. Ex. Roxb.: An Endangered Himalayan Herb

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 761
Author(s):  
Arun Kumar Khajuria ◽  
Christophe Hano ◽  
Narendra Singh Bisht
Keyword(s):  
Somatic Embryogenesis ◽  
Culture Medium ◽  
Casein Hydrolysate ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Mature Embryos ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Root Axis ◽  
Somatic Embryo Induction

Viola canescens Wall. ex. Roxb. is an important but threatened medicinal herb found at 1500–2400 m above mean sea level in the Himalayas. Overexploitation and habitat preference have put the plant under serious threat. Thus, the present study was undertaken to develop an efficient protocol for in vitro propagation via somatic embryogenesis. The results revealed that plant can be regenerated successfully through somatic embryogenesis using leaf derived calli. Regular subculturing of calli on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D)/indole-3-butyric acid (IBA)/kinetin (Kn) and varying combinations of 2,4-D+Kn induced somatic embryogenesis. The maximum average number of somatic embryos (SE) (19.15 ± 2.66) was induced on the medium with 0.15 + 0.05 mg L−1 of 2,4-D and Kn, respectively, and this medium was used as a control. To enhance somatic embryo induction, the control MS medium was supplemented with l-glutamine (200–400 mg L−1) and casein hydrolysate (1–4%). The maximum average number of SE (27.66 ± 2.67) and average mature SE (13.16 ± 3.48) were recorded on the medium having 2 % l-glutamine and 50 mg L−1 casein hydrolysate. The induced SE were asynchronous, so, to foster their maturation, the culture medium (free from growth regulators) was supplemented with abscisic acid (ABA) and silver nitrate (AgNO3). The maximum average number (35.96 ± 3.68) of mature SE was noticed on MS medium supplemented with 1.5 mg L−1 ABA. Mature embryos had two well-developed cotyledons and an elongated hypocotyl root axis. The development of SE into plantlets was significant for embryos matured on the medium with AgNO3 and ABA, with 86.67% and 83.33% conversion on the medium with 0.20 mg L−1 6-benzylaminopurine (BAP). The plantlets thus produced acclimatized in a growth chamber before being transferred to the field, which showed 89.89% survival. The plants were morphologically similar to the mother plant with successful flowering.

PLANT REGENERATION OF KAMCHATIC PLANTAIN (PLANTAGO CAMTSCHATICA LINK) THROUGH CALLUS INDUCTION

2017 ◽  
Vol 19 (3) ◽  
pp. 41-48
Author(s):  
Ay N.V. ◽  
Duy M.V. ◽  
Baatartsogt O. ◽  
Altantsetseg Kh. ◽  
Enkhchimeg V.
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Leaf Explants ◽  
Stage Iv ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Development Stage ◽  
High Survival ◽  
Ms Medium

In vitro seedling offspring of Plantago camtschatica Link was investigated regarding induction of somatic embryogenesis in petiole/leaf explants from shoot tissue and shoot proliferation. The aim of study was to investigate the medium supplemented with suitable concentration of plant growth regulators in order to induce somatic embryogenesis, plant regeneration and shoot multiplication. The results showed that: (i) Petiole/young leaf of immature stem induced the highest ratio of calli induction and compact calli formation on MS medium supplemented with 1 mgL-1 2,4-D and 0.5 mgL-1 BA; (ii) From created calli, somatic embryogenesis could be induced on MS medium supplemented with 1 mgL-1 TDZ or 1 mgL-1 TDZ and 0.5 mgL-1 NAA; (iii) MS medium supplemented with 5-7 mgL-1 BA shown the most effective on shoot development stage; (iv) Rooting of shoot was the best on 1/2 solid MS medium with activated charcoal (2 gL-1), and 0.5-4 mgL-1 NAA; and (v) acclimatization of micropropagated plants could be planted in plastic pots containing a mixture of decayed straw : rice husk ashes, (1:1, v/v), sand : soil (1:1, v/v) or soil, showed a high survival rate and most seedlings grew normally.

scholarly journals Somatic Embryogenesis and Plant Regeneration from in-vitro-grown Leaf Explants of Rose

HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1378-1380 ◽  
Author(s):  
C.K. Kim ◽  
J.Y. Oh ◽  
J.D. Chung ◽  
A.M. Burrell ◽  
D.H. Byrne
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Formation ◽  
Ms Medium ◽  
Regeneration Frequency

Somatic embryogenesis was initiated from in vitro-grown leaf explants of rose using an induction period of 4 weeks on MS basal medium supplemented with auxin followed by several subcultures on MS basal medium with cytokinin. `4th of July' showed the highest regeneration frequency (24.4%) on 5.3 μm NAA followed by culture on medium containing 18.2 μm zeatin. `Tournament of Roses' produced somatic embryos when cultured for 4 weeks on medium containing dicamba, 2.3 μm followed by three subcultures on medium containing 18.2 μm zeatin. Embryogenic callus matured on MS media containing 0.5 μm NAA, 6.8 μm zeatin, and 2.9 μm GA3. Long-term cultures were established for both cultivars. Somatic embryos germinated on MS medium containing IBA and BA. Silver nitrate (58.8 μm) enhanced shoot formation and germination of somatic embryos. Plants derived from somatic embryos were acclimatized and successfully established in the greenhouse.

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