scholarly journals Micropropagation of Asparagus densiflorus via axillary shoots, indirect organogenesis and somatic embryogenesis

2017 ◽  
Vol 29 (2) ◽  
pp. 143-153
Author(s):  
Anna Pindel

AbstractThe present study has described a simple protocol for efficient plant regeneration of Asparagus densiflorus ‘Sprengeri’ and ‘Myriocladus’ using single-node spear explants, and indirect organogenesis via callogenesis induced on internode explants. The results showed that the genotypes ‘Sprengeri’ and ‘Myriocladus’ regenerated to complete plants via nodal cultures and callus tissue, but the plant regeneration response was higher in secondary explants on MS medium with NAA + kinetin (1+1 mg dm-3) after transfer onto a multiplication medium with IAA+BAP (1+4 mg dm-3), and then onto a rooting medium supplemented with IBA (10 mg dm-3) or NAA + kinetin (1+1 mg dm-3). Primary explants of both cultivars showed high regenerative potential (via the callus stage) on MS medium with IAA+BAP. The cultivar Sprengeri also regenerated via somatic embryogenesis. Both kinds of ‘Meyeri’ explants have a morphogenetic potential for the formation of shoots, which, however, were not capable of rooting. This confirms that the explant, genotype and culture medium are determining factors in the in vitro plant regeneration system.

2012 ◽  
Vol 60 (1) ◽  
pp. 47-55 ◽  
Author(s):  
A. Ali ◽  
T. Yossef ◽  
A. El-Banna

The present study was carried out for developing an efficient in vitro callus induction and plant regeneration system in four different tomato genotypes (Solanum lycopersicum Mill., previous name: Lycopersicon esculentum), Advantage II, Edkawy, Castle Rock and Super Strain B, using hypocotyl and cotyledon explants. The effects of two cytokinins, BA (benzyl adenine) and Kin (kinetin), on callus induction and plant regeneration frequency were investigated when added to MS medium in combination at varying concentrations. All concentrations of the two cytokinins were suitable for callus induction and plant regeneration. The frequency of callus induction and plant regeneration from both cotyledon and hypocotyl explants reached 100% for all tested genotypes. Cotyledons produced a higher average number of shoots per explants than hypocotyls for all the genotypes in the five concentrations of combined cytokinins. The average number of shoots per explant in Super Strain B was found to be the highest (42 and 60 for the hypocotyl and cotyledon explants, respectively). Supplementing MS medium with 1.0 mg L−1 kinetin and 1.0 mg L−1 benzyl adenine was found to be optimum for producing the highest number of shoots per explant from hypocotyls and cotyledons in the tomato genotypes investigated. The proposed medium showed a significant superiority over the reference media.


2019 ◽  
Vol 27 ◽  
pp. 89-99
Author(s):  
M Haque ◽  
SMS Islam

This study was carried out to improve an efficient protocol for in vitro callus induction and plant regeneration using Bangladeshi barley genotypes collected from BARI, Gazipur, Bangladesh. After sterilization embryos were separated carefully from mature seeds of six barley genotypes (BB-1, BB-2, BB-3, BB-4, BB-5 and BB-6) and cultured them in MS medium supplemented with various concentration and combination of PGRs for callus induction and regeneration. Out of six genotypes BB-6 showed highest (38.17%) callus induction in MS + 4.0 mg/l 2,4-D + 200 mg/l L-proline + 300 mg/l casein hydrolysate; whereas, BB-4 and BB-5 showed no callus induction in the same medium. For plant regeneration from embryogenic calli the same genotype (BB-6) also performed the best results (19.25%) in MS medium supplemented with 1.5 mg/l BAP + 30 g/l sucrose. Analysis of variance (ANOVA) showed highly significant differences among the media and the genotypes. J. bio-sci. 27: 89-99, 2019


2022 ◽  
Author(s):  
Leticia da Silva Araújo ◽  
Virginia Silva Carvalho ◽  
Andressa Leal Generoso ◽  
Josefa Grasiela Silva Santana ◽  
Glaziele Campbell ◽  
...  

Abstract Passiflora setacea DC (Passifloraceae) is considered an important species in the genetic breeding of passion fruit. However, its use is limited due to low seed germination. This paper aimed to study the effect of cytokinins 6-benzyladenine (BA) and thidiazuron (TDZ) on the in vitro morphogenesis of P. setacea using three explants: hypocotyl, nodal segment, and root segment. The explants were induced to morphogenesis in MS medium modified and with different concentrations of BA and TDZ. After 55 days, the percentage of calluses and shoots were evaluated, and anatomical characterization was performed. The three explants used in the in vitro morphogenesis of P. setacea showed callus and shoots formation, but in greater numbers in the nodal segments treated with BA. TDZ isolated affected equal to or less than BA on callus and shoots formation for the three explants. Direct and indirect organogenesis was observed in the three types of explants. From the results obtained for plant regeneration via in vitro morphogenesis of P. setacea, it is recommended to use a nodal segment in MSM medium and supplemented with 2.22 μmol L-1 of BA.


2008 ◽  
pp. 22-33 ◽  
Author(s):  
Marilyn Belarmino ◽  
Jocelyn Gonzales

A study was conducted to establish a reliable procedure for somatic embryogenesis and plant regeneration from callus cultures of purple food yam (Dioscorea alata L.). The procedure involved three steps; (1) culture of nodal stem segments from greenhousegrown plants to generate in vitro plantlets; (2) induction of callus from the leaf, petiole and nodal stem tissues; and (3) initiation of somatic embryo from callus. Results showed that the agar-solidified Murashige and Skoog (MS) medium containing 30 gl-1 sugar, 0.1 gl-1 α-cysteine , 10 mgl-1 calcium pantothenic acid, 2.0 mgl-1 asparagine, 2.0 mgl-1 arginine, 80.0 mgl-1 adenine sulfate (AdSO4) and 0.1 mgl-1 naphthalene acetic acid (NAA) effectively broke dormancy of lateral buds of nodal stem cultures from both ‘VU-2’ and ‘Kinampay‘ varieties. Production of multiple adventitious shoots occurred after transfer of in vitro nodal pieces to the same medium added with 1.0 mgl-1 benzylamino purine (BAP) or, MSA medium. Callus was effectively induced from the vegetative tissues in MS medium added with 1.0 mgl-1 2,4-Dichlorophenoxy acetic acid (2,4-D) or, with picloram. Among the three types of explants, the nodal stem was the most suitable which produced purplish nodular embryogenic callus. A higher percentage of nodal stem-derived calli produced globular embryos in MS medium containing 1.0 mgl-1 2,4-D and 0.5 mgl-1 BAP, or in 1.0 mgl-1 picloram and 0.5 mgl-1 BAP than, in the plant growth regulator-free medium (control). The maturation of embryos was facilitated by one-month culture in MS medium containing 0.1 mgl-1 ABA and 100 mgl-1 glutamine. This step improved the germination of somatic embryos in one-half strength PGR-free MS medium containing 100 mgl-1 glutamine (regeneration medium). All somatic embryoderived plantlets were morphologically normal and established well in soil.


2021 ◽  
Vol 49 (2) ◽  
pp. 12152
Author(s):  
Yin-Kai XI ◽  
Heng-Yu HUANG

The artificial rapid propagation system for Swertia conaensis T. N. Ho et S. W. Liu was explored to screen the appropriate plant regeneration method and to provide an efficient propagation mode, useful for artificial breeding technology or for further research and development of the Tibetan endemic medicinal plant. In this study, the most suitable explant and hormone were chosen according to single factor test. Next, the effects of different hormone combinations on basal stem cluster bud induction, callus induction, adventitious bud occurrence and plant regeneration were investigated by using complete combination and orthogonal experiment. The obtained results showed that the explants suitable for in vitro of S. conaensis were stem tips with leaves, which were regenerated through the method of basal stem cluster bud occurrence in the MS medium with 2.0 mg∙L-1 6-BA, 0.5 mg∙L-1 NAA, but the proliferation coefficient was low, only 3.16 after 40 days of culture. Subsequently, the proliferation coefficient failed to improve, irrespective of change of the concentration ratio of 6-BA and NAA. Therefore, in the orthogonal experiment of adding ZT, the MS medium with 1.0 mg∙L-1 ZT, 0.5 mg∙L-1 NAA and 2.5 mg∙L-1 6-BA induced a large number of callus green and compact, with 86.30% callus occurrence rate. After 40 days of culture, the rate of adventitious bud occurrence was 96.55% and the proliferation coefficient was high (10.37). The rooting rate was 100% in the 1/2MS medium with 0.5 mg∙L-1 NAA. The survival rate of regenerated plants was more than 95%. Indirect organogenesis was more efficient than direct organogenesis in in vitro culture of S. conaensis. In this study, the efficient and stable regeneration system of S. conaensis was achieved through the method of explant to callus to adventitious buds, which provided an effective way to an endangered species.


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.


Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna

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.


Author(s):  
D. Bele ◽  
Nishi Mishra ◽  
Sushma Tiwari ◽  
M. K. Tripathi ◽  
G. Tiwari

Nodal segments of sandalwood were cultured on MS medium amended with different plant growth regulators in varying concentrations to search out higher in vitro response leading to plantlet regeneration via somatic embryogenesis and/or organogenesis. Higher proportion of direct somatic embryogenesis, number(s) of somatic embryo per explant and plantlet regeneration via direct organogenesis were recorded on MS medium supplemented with a moderate concentration of TDZ (1.0 mgl-1) in combination with comparatively a lower concentration of NAA (0.5 mgl-1). A relative higher concentration of BAP (1.0-2.0 mgl-1) in combination with a lower concentration of NAA (0.5 mgl-1) promoted frequency of indirect somatic embryogenesis. Ratio of organ formation directly from surface of cultured explants was recovered from culture medium fortified with a higher concentration of BA at the concentration of 4.0 mgl-1 in combination with a lower concentration of NAA (0.5 mgl-1). Maximum plantlets regenerated via somatic embryogenesis (direct and/or indirect) on regeneration medium supplemented with 2.0 mgl-1TDZ  in combination with 1.0 mg l-1GA3, while plantlets in higher frequencies via indirect organogenesis was attained with regeneration medium amended with comparatively lower concentration of TDZ (1.0 mg l-1) in combination with 0.5 mgl-1 GA3 and 0.5     mgl-1 NAA. The plantlets were transferred to pots and hardened in Environmental Growth Cabinet and Net House during preliminary weaning period and transferred to field successfully. Morphologically normal plants were recovered.


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny

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.


2017 ◽  
Vol 27 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Mustafa Abul Kalam Azad ◽  
Muhammad Nurul Amin

A plant regeneration system was established from hypocotyl explants of in vitro grown seedlings of A. officinalis and in vitro proliferated shoots, respectively through somatic embryogenesis and embryogenic calli. Somatic embryogenesis was significantly influenced by the types of plant growth regulators. Embryogenic calli with somatic embryos developed well in MS supplemented with 2.0 ‐ 4.0 μM BAP and 1.0 ‐ 4.0 μM 22,4‐D, NAA or IBA. The highest frequency (95.3%) of embryogenic calli and 55.2 somatic embryos formation were obtained when the MS was amended with 4.0 μM BAP and 2.0 μM 2,4‐D. The best embryo germination occurred in 1.0 μM BAP supplemented MMS. The highest 97.2% of shoot proliferation was observed in embryogenic calli in MS medium containing 2.0 μM BAP and 1.0 μM IBA. In vitro grown shoots were rooted in MMS with 1.0 ‐ 2.0 μM IBA. Regenerants were transferred to vermicompost and successfully established under an ex vitro environment in garden soil with 80% survival rate.Plant Tissue Cult. & Biotech. 27(1): 21-31, 2017 (June)


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