scholarly journals Phloroglucinol Mediated Plant Regeneration of Ornithogalum dubium as the Sole “Hormone-Like Supplement” in Plant Tissue Culture Long-Term Experiments

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 929
Author(s):  
Carloalberto Petti

Tissue culture is an essential requirement in plant science to preserve genetic resources and to expand naturally occurring germplasm. A variety of naturally occurring and synthetic hormones are available to induce the processes of dedifferentiation and redifferentiation. Not all plant material is susceptible to tissue culture, and often complex media and hormone requirements are needed to achieve successful plant propagations. The availability of new hormones or chemicals acting as hormones are critical to the expansion of tissue culture potentials. Phloroglucinol has been shown to have certain hormone-like properties in a variety of studies. Ornithogalum dubium, an important geophyte species, was used to characterise the potential of phloroglucinol as the sole plant-like hormone in a tissue culture experiment. Tissue culture, plant regeneration, total phenolic and genetic variability were established by applying a variety of methods throughout long-term experiments. Phloroglucinol did induce callus formation and plant regeneration when used as the sole supplement in the media at a rate of 37%, thus demonstrating auxin/cytokines-like properties. Callus formation was of 3 types, friable and cellular, hard and compact, and a mixture of the two. The important finding was that direct somatogenesis did occur albeit more frequently on younger tissue, whereby rates of induction were up to 52%. It is concluded that phloroglucinol acts as a “hormone-like” molecule and can trigger direct embryogenesis without callus formation.

2021 ◽  
Vol 9 (2) ◽  
pp. 109-115
Author(s):  
Ahmad Saifun Naser ◽  
Muhammad Wisnu

Availability of quality seeds in production of krisan (Chrysanthemum morifolium Ramat cv Dewi ratih) cultivation is still rare, therefore research on seed multiplication through tissue culture is needed. The media used in tissue culture is relatively expensive for home industry. This study aims to determine the respond of leaf and stem explants using foliar fertilizers (Growmore, Gandasil D and Mutiara) as an alternative media for callus inductions. This study used a Completely Randomized Design (CRD) consisted of 4 treatments: P0: ½ MS + 0,25 mg/l BAP, P1 (Growmore + 0,25 mg/l BAP), P2 (Gandasil D + 0,25 mg/l BAP), P3 (Mutiara + 0,25 mg/l BAP). The variables observed in this study included callus appearance time, callus color and callus texture. The result of this study indicated that the use of BAP (6-Benzyl Amino Purine) affected the time of callus formation and callus morphology. Callus was formed on leaf explants 13 days after planting while on stem explants 7 days after planting and compact texture. Growmore + 0,25 mg/l BAP treatment yields the best callus on leaf explant, while Gandasil D + 0,25 mg/l BAP treatment yields the best callus on stem explant.


2014 ◽  
Vol 644-650 ◽  
pp. 5407-5410
Author(s):  
Hui Fang Chi

s. The cotyledons, Internodes, leaves and stems of sweet broad pea were studied on tissue culture. Research results show that: The ability of different explants for callus formation and adventitious bud differentiation in different culture medium is different. The callus formation rate and sprouting rate of Internodes is significantly higher than other explants, which is a ideal material for tissue culture. The callus formation rate of Internodes was 100% in MS +BA1.0 mg/L+NAA 1.0 mg/L and MS+ 2, 4-D 0.5 mg/L; The bud differentiation is best at the medium of MS+ 6-BA 2 mg/L, which reached 86.7%; the rooting rate was 83.3% at the medium of MS+ NAA 3mg/L. The study provides a experimental basis for further study on the plant regeneration in the sweet broad pea.


2015 ◽  
Vol 24 (2) ◽  
pp. 213-221 ◽  
Author(s):  
Ahmad H Al Gabbiesh ◽  
M Ghabeish ◽  
I H ◽  
M Kleinwächter ◽  
D Selmar

Somatic embryogenesis was induced in embryo culture on half MS medium supplemented with NAA (8 mg/l) as the sole plant growth regulator after incubation of the media in the refrigerator at 4°C for two weeks to promote callus induction and somatic embryogenesis in Laurus nobilis. Both embryogenetic calli and somatic embryos were induced in the above selected medium. Embryo growth and development were stimulated by separation of embryos successfully from embryo clusters and transferred onto fresh half MS. Among the selected explants, only leaf bases were found to respond actively to plant regeneration, especially in inducing callus formation and in sustaining faster callus growth. Root formation of regenerated plantlets tended to decrease with time on regeneration media. Overall, 75% of the plantlets derived from the callus survived in the greenhouse; and they all grew to phenotypically normal plants. This procedure will enable the use of regeneration tissue culture technology for germplasm conservation of L. nobilis, a plant of high medicinal and commercial value.Plant Tissue Cult. & Biotech. 24(2): 213-221, 2014 (December)


2020 ◽  
Vol 71 (21) ◽  
pp. 6789-6798 ◽  
Author(s):  
Henrique Noronha ◽  
Angélica Silva ◽  
Namiki Mitani-Ueno ◽  
Carlos Conde ◽  
Farzana Sabir ◽  
...  

Abstract Silicon (Si) supplementation has been shown to improve plant tolerance to different stresses, and its accumulation in the aerial organs is mediated by NIP2;1 aquaporins (Lsi channels) and Lsi2-type exporters in roots. In the present study, we tested the hypothesis that grapevine expresses a functional NIP2;1 that accounts for root Si uptake and, eventually, Si accumulation in leaves. Own-rooted grapevine cuttings of the cultivar Vinhão accumulated >0.2% Si (DW) in leaves when irrigated with 1.5 mM Si for 1 month, while Si was undetected in control leaves. Real-time PCR showed that VvNIP2;1 was highly expressed in roots and in green berries. The transient transformation of tobacco leaf epidermal cells mediated by Agrobacterium tumefaciens confirmed VvNIP2;1 localization at the plasma membrane. Transport experiments in oocytes showed that VvNIP2;1 mediates Si and arsenite uptake, whereas permeability studies revealed that VvNIP2;1 expressed in yeast is unable to transport water and glycerol. Si supplementation to pigmented grape cultured cells (cv. Gamay Freáux) had no impact on the total phenolic and anthocyanin content, or on the growth rate and VvNIP2;1 expression. Long-term experiments should help determine the extent of Si uptake over time and whether grapevine can benefit from Si fertilization.


Plants ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 79
Author(s):  
Sitong Liu ◽  
Xinfeng Wang ◽  
Ning Ding ◽  
Yutong Liu ◽  
Ning Li ◽  
...  

Plants have the salient biological property of totipotency, i.e., the capacity to regenerate a whole plant from virtually any kind of fully differentiated somatic cells after a process of dedifferentiation. This property has been well-documented by successful plant regeneration from tissue cultures of diverse plant species. However, the accumulation of somaclonal variation, especially karyotype alteration, during the tissue culture process compromises cell totipotency. In this respect, Chinese ginseng (Panax ginseng C. A. Mey.) is an exception in that it shows little decline in cell totipotency accompanied by remarkable chromosomal stability even after prolonged tissue cultures. However, it remains unclear whether chromosomal level stability necessarily couples with molecular genetic stability at the nucleotide sequence level, given that the two types of stabilities are generated by largely distinct mechanisms. Here, we addressed this issue by genome-wide comparisons at the single-base resolution of long-term tissue culture-regenerated P. ginseng plants. We identified abundant single nucleotide polymorphisms (SNPs) that have accumulated in cultured ginseng callus and are retained in the process of plant regeneration. These SNPs did not occur at random but showed differences among chromosomes and biased regional aggregation along a given chromosome. In addition, our results demonstrate that, compared with the overall genes, genes related to processes of cell totipotency and chromosomal stability possess lower mutation rates at both coding and flanking regions. In addition, collectively, the mutated genes exhibited higher expression levels than non-mutated genes and are significantly enriched in fundamental biological processes, including cellular component organization, development, and reproduction. These attributes suggest that the precipitated molecular level genetic variations during the process of regeneration in P. ginseng are likely under selection to fortify normal development. As such, they likely did not undermine chromosomal stability and totipotency of the long-term ginseng cultures.


Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Yeong Yeop Jeong ◽  
Hun-Young Lee ◽  
Suk Weon Kim ◽  
Yoo-Sun Noh ◽  
Pil Joon Seo

Abstract Background Plants have a remarkable reprogramming potential, which facilitates plant regeneration, especially from a single cell. Protoplasts have the ability to form a cell wall and undergo cell division, allowing whole plant regeneration. With the growing need for protoplast regeneration in genetic engineering and genome editing, fundamental studies that enhance our understanding of cell cycle re-entry, pluripotency acquisition, and de novo tissue regeneration are essential. To conduct these studies, a reproducible and efficient protoplast regeneration method using model plants is necessary. Results Here, we optimized cell and tissue culture methods for improving protoplast regeneration efficiency in Arabidopsis thaliana. Protoplasts were isolated from whole seedlings of four different Arabidopsis ecotypes including Columbia (Col-0), Wassilewskija (Ws-2), Nossen (No-0), and HR (HR-10). Among these ecotypes, Ws-2 showed the highest potential for protoplast regeneration. A modified thin alginate layer was applied to the protoplast culture at an optimal density of 1 × 106 protoplasts/mL. Following callus formation and de novo shoot regeneration, the regenerated inflorescence stems were used for de novo root organogenesis. The entire protoplast regeneration process was completed within 15 weeks. The in vitro regenerated plants were fertile and produced morphologically normal progenies. Conclusion The cell and tissue culture system optimized in this study for protoplast regeneration is efficient and reproducible. This method of Arabidopsis protoplast regeneration can be used for fundamental studies on pluripotency establishment and de novo tissue regeneration.


1979 ◽  
Vol 90 (3) ◽  
pp. 421-433 ◽  
Author(s):  
Loren G. Lipson ◽  
Inese Z. Beitins ◽  
Paul L. Kornblith ◽  
Janet W. McArthur ◽  
Henry G. Friesen ◽  
...  

ABSTRACT A study was undertaken to determine the length of time that human pituitary tumours are capable of releasing anterior pituitary polypeptide hormones in vitro under basal conditions and to study the spectrum of hormone release by functioning and "non-functioning" pituitary neoplasms. Fragments from the pituitary tumours of 10 patients in the following categories: 1 Cushing's disease, 2 with amenorrhoea-galactorrhoea, 3 with acromegaly, and 4 with "non-functioning" pituitary tumours and from 2 normal human anterior pituitary glands were placed in primary culture immediately after surgery. The in vitro release of human growth hormone (hGH), prolactin (Prl), thyrotrophin (TSH), adrenocorticotrophin (ACTH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) was measured by specific radioimmunoassays at the end of each week in culture. Hormone release was surveyed from 6 weeks to 6 months depending upon the survival of the culture. Hormone release patterns were compared with clinical and pathological data. In the initial week of the study, all 6 anterior pituitary polypeptides were detected in the media from the 2 control pituitaries and from 4 of the tumours (1 amenorrhoea-galactorrhoea and 3 acromegaly) in concentrations up to 100 ng/ml of medium while 5 of the 6 hormones were readily detectable in the media from 2 additional tumour samples (Cushing's disease and 1 "non-functioning" pituitary tumour). The media of the remaining 4 tumours contained at least 3 of the 6 hormones (1 amenorrhoea-galactorrhoea and 3 "non-functioning" pituitary tumours). After 6 months in culture, the 6 hormones were readily detectable in at least 1 of the 5 surviving cultures and hGH (up to 800 ng/ml) and LH were each detectable in the media from 2 cultures. Although most of the hormone concentrations in the media decreased with length of time in culture, there were 2 exceptions. First, in the media from 5 of the 12 cultures from both controls and tumours, Prl concentrations increased after 50 to 80 days culture. This increase usually lasted for several weeks before Prl levels again began to decline. The second unusual finding occurred in a tumour from a patient with acromegaly in the media of which hGH levels rose from 60 ng/ml to 800 ng/ml between days 125 and 174. These findings of prolonged hormone release in vitro give promise of future usefulness of tissue culture methods for study of polypeptide hormone releasing mechanisms and long-term production of human anterior pituitary hormones for use in research and possible therapy.


HortScience ◽  
1994 ◽  
Vol 29 (8) ◽  
pp. 906-909 ◽  
Author(s):  
H. Lou ◽  
S. Kako

The embryogenic capacity of seven cucumber (Cucumis sativus L.) cultivars was examined by tissue culture of cotyledon, young first-leaf, and internode explants. Somatic embryogenesis frequencies differed significantly among the tested cultivars, and `Fushinarimidori' produced the highest number of embryos from either cotyledons or young first leaves. Cotyledon- and first-leaf-derived calluses produced more embryos than calluses from internodes. Somatic embryos were induced from `Aonaga F1' internodes. With relatively high sucrose levels (6% and 9%) in the initiation medium, the frequency of embryogenic callus formation from `Fushinarimidori' cotyledon explants was >90%. The highest yield of somatic embryos occurred in cultures initiated with high sucrose levels (9% or 12%), although 12% sucrose inhibited callus formation and growth. Somatic embryos germinated in a basal liquid medium supplemented with 0.5% activated charcoal, and they developed into well-shaped, healthy plantlets on semisolid medium with 1% sucrose.


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