scholarly journals Induction of Callogenesis, Organogenesis, and Embryogenesis in Non-Meristematic Explants of Bleeding Heart and Evaluation of Chemical Diversity of Key Metabolites from Callus

2020 ◽  
Vol 21 (16) ◽  
pp. 5826
Author(s):  
Dariusz Kulus ◽  
Alicja Tymoszuk

Lamprocapnos spectabilis (L.) Fukuhara is a perennial plant species valued in the horticultural, cosmetic, and pharmaceutical markets. To date, however, there were no studies on tissue culture systems in this species when adjusted from non-meristematic explants. The aim of this study is to induce callogenesis, organogenesis, and somatic embryogenesis in non-meristematic explants of Lamprocapnos spectabilis ‘Alba’ cultured in various media and to analyze the chemical diversity of the produced callus. Leaf, petiole, and internode explants were cultured on the modified Murashige and Skoog (MS) medium fortified with various combinations and concentrations of 6-benzyladenine (BA), indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorphenoxyacetic acid (2,4-D), and picloram (PIC). After 10 weeks of culturing, the morphogenetic response of explants was evaluated and the concentration of chlorophylls, carotenoids, anthocyanins, and polyphenols in callus was analyzed. There was no influence of explant type on the callogenesis efficiency (62.1–65.3%). The highest fresh weight of callus was produced on leaf explants in the presence of 2,4-D or PIC. In contrast, the highest share of dry weight was found in internode-derived calli and cultured on IAA-supplemented medium (up to 30.8%). Only 2.5% of all explants regenerated adventitious shoots, while rhizogenesis was reported in 4.5% of explants. Somatic embryos were produced indirectly by 0% to 100% of explants, depending on the culture medium and explant type. The highest mean number of embryos (11.4 per explant) was found on petioles cultured in the MS medium with 0.5 mg·L−1 BA and 1.0 mg·L−1 PIC. Calli cultured in media with NAA usually contained a higher content of primary and secondary metabolites. There was also a significant impact of explant type on the content of anthocyanins, polyphenols, and carotenoids in callus. Further studies should focus on the elicitation of metabolites production in callus culture systems of the bleeding heart.

HortScience ◽  
2017 ◽  
Vol 52 (9) ◽  
pp. 1278-1282 ◽  
Author(s):  
Boling Liu ◽  
Hongzhou Fang ◽  
Chaorong Meng ◽  
Ming Chen ◽  
Qingdong Chai ◽  
...  

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.


2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
K. Varutharaju ◽  
C. Soundar Raju ◽  
C. Thilip ◽  
A. Aslam ◽  
A. Shajahan

An efficient protocol for direct shoot organogenesis has been developed for the medicinal plantAerva lanata(L.) Juss. ex Schult. Regeneration was achieved from leaf segments of 20 days oldin vitroplantlets raised on Murashige and Skoog (MS) medium containing 0.25–2.0 mg L−1thiadiazuron (TDZ), 3% sucrose, and 0.8% agar. After 21 days of culture incubation, maximum number of shoot organogenesis (23.6 ± 0.16) was obtained on medium containing 1.0 mg L−1TDZ. The shoots were able to producein vitroflowers on medium containing 1.0 mg L−1TDZ in combination with 0.25–0.5 mg L−1  α-naphthaleneacetic acid (NAA). Histological observation showed that the epidermal cells of the leaf explants exhibited continuous cell division led to formation of numerous dome shaped meristematic protrusions and subsequently developed into adventitious shoots. Upon transfer of shootlets to half strength MS medium containing 1.0 mg L−1indole-3-butyric acid (IBA), around 86% of the regenerated shoots formed roots and plantlets. Rooted plants were hardened and successfully established in the soil at the survival rate of 92%. The regeneration protocol developed in this study provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for a large scale production of its medicinally active compounds and genetic transformations for further improvement.


HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1373-1377 ◽  
Author(s):  
Margarita Fraga ◽  
Mertxe Alonso ◽  
Marisé Borja

Meristem culture and/or thermotherapy were used for virus elimination from ornamental Phlox paniculata L. (`Blue Boy', `Orange perfection' and `Starfire') mother plants. Shoot tip, leaf, node and flower ovary explants collected from greenhouse-maintained virus free plants were cultured in vitro for shoot initiation. Adventitious shoot initiation was observed on Murashige and Skoog (MS) medium containing the cytokinin BA with or without the auxin NAA. The addition of 0.4 mg·L-1 thiamine, 0.4 mg·L-1 folic acid, and 40 mg·L-1 adenine sulfate to the MS medium did not improve the regeneration rate. Multiplication and rooting were genotype dependent. Blue Boy and Orange Perfection cultivars regenerated the maximum number of shoots from leaf explants. `Blue Boy' leaf explants from in vitro plants had a lower regeneration rate than explants from greenhouse plants. Cultivar `Starfire' had the highest shoot formation with open flower ovary explants and failed to regenerate from leaf explants. In vitro rooting of adventitious shoots in the presence of auxins (IAA, NAA, or IBA) with or without BA was less effective than ex vitro rooting. Chemical names used: 6-benzyladenine (BA); indole-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).


HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1050E-1051
Author(s):  
Wenhao Dai ◽  
Victoria Jacques

Periwinkle, a perennial commonly used as a summer bedding plant, is known as the source of vinca alkaloids used to treat lymphocytic leukemia and Hodgkin's disease. It is also one of the natural hosts of many phytoplasma diseases, such as X-disease of major Prunus species, aster yellows, and ash yellows diseases. Therefore, periwinkle is an ideal plant species for phytoplasma disease research, such as disease transmission, species resistance, and resistant gene screening. Periwinkle tissue culture was established by incubating sterile seeds in hormone-free Murashige and Skoog (MS) medium. Plants were successfully regenerated from in vitro leaf tissues of periwinkle. Adventitious shoots were induced when leaf tissues were cultured on Murashige and Skoog (MS) medium or woody plant medium (WPM) supplemented with benzyladenine (BA) and naphthaleneacetic acid (NAA). Nearly 75% of leaf explants produced shoots in both media with 10–20 μm BA and 1 μm NAA. A mean of 4.3 shoots was produced from each explant cultured on WPM, whereas only 2 shoots were produced on MS medium under 16-h photoperiod. Leaf explants under dark treatment for 2 weeks produced big callus only, indicating that light is necessary for shoot formation. Most adventitious shoots were induced from the joint of leaf blade and petiole. In vitro shoots (>1.5 cm) were easily rooted in half-strength MS medium. Addition of NAA dramatically increased root number, with more than 20 roots being induced in 5 μm NAA medium. Rooted plants were transferred to potting medium and grown in a greenhouse.


HortScience ◽  
1994 ◽  
Vol 29 (11) ◽  
pp. 1353-1354
Author(s):  
Chiwon W. Lee ◽  
Joel T. Nichols ◽  
Lijuan Wang ◽  
Shanqiang Ke

Excised leaf sections of lance coreopsis cultured on Murashige Skoog (MS) medium produced adventitious shoots in response to BA. When the combinations of 0, 0.5, 1, or 2 μm NAA with 0, 5, 10, 20, or 40 μm BA were tested, shoots were induced by any of the four BA concentrations used in the medium, regardless of the presence of NAA. The average number of shoots formed per leaf section ranged from 1.4 to 4.3 seven weeks after culture initiation. Roots were induced at the base of individual shoots on the same regeneration medium when cultures were kept longer than 7 weeks. The rooted plants were transferred successfully into soil. The regenerated plants had the same growth and flowering characteristics as the seed-grown plants. Chemical names used: benzyladenine (BA); naphthaleneacetic acid (NAA).


HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Ana Carolina Espinosa ◽  
Paula M. Pijut ◽  
Charles H. Michler

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μm 6-benzylaminopurine (BA), 0.49 μm indole-3-butyric acid (IBA), and 0.29 μm gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μm thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μm naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μm BA in combination with 0, 0.54, 1.07, or 5.37 μm NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μm TDZ plus 0.54 μm NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μm TDZ plus 1.07 μm NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μm IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μm IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.


2012 ◽  
Vol 81 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Jun Li ◽  
Ye Kang ◽  
Sheng Qiang ◽  
Gary Peng

Goldenrod (<em>Solidago canadensis </em>L.) is an invasive plant species in many countries except North America but a cut-flower species worldwide. There is a need to generate and propagate goldenrod clones efficiently for research and commercial purposes. A callus induction and plantlet regeneration system was developed by studying the influence of explant type and different concentrations of plant growth regulators. The highest callus production from leaf segments was obtained on Murashige and Skoog’s medium (MS medium) supplemented with 1.0 mg/L naphthalene acetic acid (NAA) and 1.0 mg/L 6-benzylaminopurine (BA). Adventitious shoots could be regenerated directly from leaf explants without an intermediate callus phase with the highest shoot induction percentage of 87.2%. The largest number of adventitious shoots per leaf explant (3.2) was obtained on MS medium supplemented with 0.4 mg/L NAA and 2.0 mg/L BA. MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L BA was the best medium for axillary shoot regeneration from nodal segments. The highest root number and longest roots occurred on half-strength MS without the addition of any growth regulator. Rooted plantlets were then transferred to a soil-based growth medium, placed in a greenhouse, and acclimatized with 100% success. All surviving plants grew normally without showing any morphological varia&shy;tion when compared to those grow from seed. This regeneration protocol may be used to produce certain biotypes of goldenrod suitable for genetic transformation rapid propagation of goldenrod for commercial purposes or for screening fungi and toxins as potential biocontrol agents against this weed.


2020 ◽  
Vol 5 (1) ◽  
pp. 75-84
Author(s):  
Seied Mehdi Miri

AbstractThe present study describes a protocol for micro-propagation, callus induction, and shoot regeneration of ginger (Zingiber officinale). The rhizomes were surface-sterilized with ethanol (70%) for 45 s, sodium hypochlorite (2.5%) for 10 min, and mercuric chloride (0.1%) for 10 min. Multiple shoots were induced from sprouting bud explants cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) combined with kinetin (Kin). The maximum shoot number was obtained from MS medium containing 10 mg/l BA with a mean of 20.6 shoots per explant. The leaf explants were cultured on MS medium supplemented with indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), Dicamba, or BA for callus culture. Green-red compact calli were induced using 2,4-D, Dicamba or BA. Also, BA successfully induced plant regeneration. The multiplied shoots that were transferred to the rooting medium (½MS supplemented with 0, 1 and 2 mg/l IAA, indole-3-butyric acid (IBA) or NAA) showed development of roots (100%). The rooted plantlets were transferred to pots containing a 1:1 mixture of cocopeat and perlite, and acclimatization was successful, resulting in 85% survival of the plantlets in the greenhouse.


Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.


HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 460B-460
Author(s):  
David J. Williams ◽  
Karim H. Al-Juboory

The objective of this study was to evaluate the ability of various cultivars of Hosta ovary explants to generate adventitious shoots and obtain variegated plants in vitro. Immature inflorescences along with 8 to 10 cm of scape were harvested from Hosta cultivars. The ovaries were prepared for culture by cutting immature florets before anthesis. The florets were first cut just above the top of the immature ovary to remove the sigma, style, corolla, and anther. Then the calyx and filament bases were also removed. Ovaries were transversely cut into halves and transferred to baby jars containing Hosta initiation medium supplemented with naphthaleneacetic acid (NAA) at 0.5 mg/L and 6-benzylamino purine (BA). The explants produced adventitious shoots from ovary base via organogenesis. The number of shoots regenerated from shoot tips and callus increased linearly with repeatedf subculturing on MS medium. This method would provide an effective alternative to conventional propagation crown division of Hosta, an expensive and slow process. The long-term goal of this project is to improve Hosta.


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