Regeneration of whole plants from cultured root tips of Limnophila indica

1981 ◽  
Vol 59 (6) ◽  
pp. 969-973 ◽  
Author(s):  
Sunanda Rao ◽  
H. Y. Mohan Ram

Root tips (1 cm long) were excised from in vitro grown plants of Limnophila indica (L.) Druce and cultured in liquid medium with 2% sucrose. The explants floated horizontally on the surface of the medium; they became green, and their tips started to grow geotropically into the medium. Four to six shoot buds each were differentiated in 84.5% of the cultures in 4 weeks. Subsequently, shoot buds covered the surface of the bent part of the root and portions above and below it. No buds were formed at the cut end or near the growing root tip. Buds failed to differentiate in roots in which the apex was not growing. Histological studies of the roots showed that the buds originated from proliferating cortical cells, which had prominent chloroplasts and starch grains. The shoot buds grew into entire plants which flowered eventually.

1972 ◽  
Vol 50 (5) ◽  
pp. 1091-1096 ◽  
Author(s):  
S. Nemec

Host–parasite interactions between eight Pythium species, previously isolated from necrotic strawberry root tissue, and Fragaria × ananassa Duch. runner plant main roots were studied. When excised roots were placed in water on petri dish cultures, P. rostratum and P. hypogenum infection was limited to the root tip epidermis. Pythium irregulare, P. perniciosum, P. dissotocum, P. sylvaticum, P. ultimum, and unidentified isolate 1E penetrated the epidermis several centimeters along the zone of maturation. Penetration occurred most frequently through root hairs by hyphae; resulting infections usually stopped in the epidermis or adjacent cortex, and here hyphae formed sporangium-like structures, oogonia, and oospores. Extensive cortical infection by hyphae was primarily limited to the zone of elongation. Extent of epidermal and cortical infection in roots grown in soil with P. irregulare, P. dissotocum, and P. sylvaticum was similar to petri dish test results. Stelar infection by P. sylvaticum and P. dissotocum occurred mainly in phloem cells, but did not advance far into the region of maturation. Numerous oogonia formed in infected steles and usually in adjacent cortical cells. Infected roots ranged from a white to grey, water-soaked color. Extensively infected root tips typically were unhealthy, succulent, and thin.


1979 ◽  
Vol 178 (3) ◽  
pp. 661-671 ◽  
Author(s):  
J R Green ◽  
D H Northcote

Two types of experiments were carried out; either maize roots were incubated in L-[1-3H]fucose or membranes were prepared from root tips and these were incubated with GDP-L-[U-14C]fucose or UDP-D-[U-4C]glucose. The radioactively labelled lipids that were synthesized in vivo and in vitro were extracted and separated into polar and neutral components. The polar lipids had the characteristics of polyprenyl phosphate and diphosphate fucose or glucose derivatives, and the neutral lipids of sterol glycosides (fucose or glucose). A partial separation of the glycolipid synthetase reactions was achieved. Membranes were fractionated into material that sedimented at 20,000g and 100,000g. Most of the polar glycolipid synthetase activity (for the incorporation of both fucose and glucose) was located in the 100,000 g pellet, and this activity was probably located in the endoplasmic reticulum. The neutral lipid, which contained fucose, was synthesized mainly by membranes of the 20,000g pellet, and the activity was probably associated with the dictyosomes, whereas the neutral glucolipids were synthesized by all the membrane fractions. It is suggested that the polar (polyprenyl) lipids labelled with fucose could act as possible intermediates during the synthesis of the glycoproteins and slime in the root tip.


2021 ◽  
Vol 13 (3) ◽  
pp. 10995
Author(s):  
Wei-Seng HO ◽  
Wee-Hiang ENG ◽  
Kwong-Hung LING

Chromosome count is the only direct way to determine the number of chromosomes of a species. This study is often considered trivial that seldom described and discussed in detail. Therefore, it is inevitable that the chromosome count protocol should be revised and revisited before it becomes obliterated. In the present study, we encountered challenges in obtaining a clear micrograph for the chromosome count of active mitotic cells of Neolamarckia cadamba (Roxb.) Bosser (Rubiaceae) root tips. Several obstacles were determined through micrograph observation, such as existing unwanted particles in cells, poor chromosome staining and chromosome clumping. To overcome these, root tip types, staining methodologies, squashing methods were among the factors assessed to obtain clear micrographs. The chromosome counts of N. cadamba under optimized procedure showed 2n = 44 chromosomes. We also apply digital technology in chromosome counts, such as online databases and graphic software that are open source and freely accessible to the public. Only basic laboratory equipment and chemicals were used throughout the study, thus making this study economical and applicable in a basic laboratory. The availability of online digital software and databases provide open-source platforms that will ease the efforts in chromosome count.


2019 ◽  
Vol 34 (1) ◽  
pp. 61-70
Author(s):  
Ana Carvalho ◽  
Fernanda Leal ◽  
Manuela Matos ◽  
José Lima-Brito

The degree of tolerance to heat stress (HS) differs among grapevine varieties. HS affects the duration and the regularity of the cell cycle in plants. The cytogenetic studies in grapevine are scarce, and the consequences of HS in the mitosis are barely known. This work intends to evaluate the consequences of induced HS in the mitotic cell cycle and chromosomes of four wine-producing varieties: Touriga Franca (TF), Touriga Nacional (TN), Rabigato and Viosinho using a cytogenetic approach. HS (1h at 42 ºC) was induced in plants of the four grapevine varieties that grew in vitro for 11 months. Plants of the same varieties and with equal age were used as control (maintained in vitro at 25 ºC). Three plants per variety and treatment (control and HS) were analysed. After HS, root-tips were collected in all plants and immediately fixed to be used for the preparation of mitotic chromosome spreads. In total, 6,116 root-tip cells were scored. Among the 5,973 dividing cells, 24.33% showed different types of irregularities in all mitotic phases (prophase, metaphase, anaphase, and telophase). After HS, the mitotic index (MI) decreased in the varieties TF and Viosinho, and increased in TN and Rabigato, relatively to the control. However, the average values of MI did not show statistically significant differences (p ˃ 0.05) among varieties, treatments and for the variety x treatment interaction. The percentage of dividing cells with anomalies (%DCA) increased after HS in all varieties relatively to the control. The average values of %DCA presented statistically significant differences (p < 0.05) only between treatments. As far as we know, this work constitutes the first cytogenetic evaluation of the HS effects in the mitotic cell cycle and chromosomes of grapevine using meristematic cells of root-tips. TN has been considered tolerant to various abiotic stresses (drought and excessive heat and light) based on other methodologies. TF and Viosinho have been referred as tolerant to abiotic stress without deeper studies available, and till the development of our investigation, the sensibility of Rabigato was unknown. Among the varieties analysed in this work, Rabigato revealed to be the less tolerant to HS. This research can be useful for selection of grapevine clones more tolerant to HS for commercialization and for the improvement of the economic sustainability.


2016 ◽  
Vol 6 (2) ◽  
pp. 65
Author(s):  
Ifa Manzila ◽  
Sri H Hidayat ◽  
Ika Mariska ◽  
Sriani Sujiprihati

<p>Callus Induction and Regeneration of Shoot and Root of<br />Chill through In Vitro Culture. Ifa Manzila, Sri H.<br />Hidayat, Ika Mariska, and Sriani Sujiprihati. In vitro<br />culture is one way for a fast and effective plant propagation.<br />This method is also useful for preliminary selection of plant<br />resistance to disease, including the chili. In vitro propagation<br />method for chili has not been widely reported. A study was<br />conducted to obtain effective techniques for callus induction<br />and regeneration into shoots on three red chili cultivars (cv)<br />Gelora, Sudra, and Chili 109. The study consisted of four<br />activities, namely the induction of callus formation,<br />induction of embryogenic callus, callus regeneration into<br />adventitious shoots, and root induction from the adventitious<br />shoots. Murashige Skoog (MS) medium + 0.6% agar + 3%<br />sucrose were used as basal medium, 20 ml/bottle. Young<br />leaves, hypocotyls and root tips of 21-day-old chili seedlings<br />were used as sources of explants. Each experiment was<br />arranged in a completely randomized design with 10<br />replications, one culture bottle for each treatment. The<br />callus induction experiments using the explants of young<br />leaf explants, hypocotyl, and root tips were done separately.<br />Each treatment consisted of explants from the three chili<br />cultivars on MS medium containing three composition of<br />growth regulators (PGR) BAP + NAA, 10 explants/bottle. The<br />embryogenic callus induction was conducted by growing<br />the callus in bottles containing a medium that contains three<br />compositions PGR 2,4-D + thidiazuron 0.5 mg/l. Induction of<br />shoot formation was done by growing the embryogenic<br />callus on medium containing three composition of plant<br />growth regulator BAP + NAA. Induction of root formation<br />was performed by growing adventitious shoots on ½ MS and<br />1 MS medium + NAA 0.5 to 1.0 mg/l. The results showed that<br />young leaves are the best explant source for callus and<br />shoot formations in chili through tissue culture compared<br />with the hypocotyl and the tip. Gelora is the most responsive<br />chili cultivar to callus, shoots, and roots formation of in their<br />respective medium, compared to Sudra and Chile 109. MS<br />medium containing BAP 3-7 mg/ml and NAA 1 mg/ml can be<br />used to induce the growth of callus from young leaf<br />explants, hypocotyl and seedling root tip chili cv Gelora,<br />Sudra, and Chile 109, but its growth was very slow and did<br />not produce embryogenic callus. Embryogenic callus<br />formation can be induced by both non-embryogenic callus<br />Hak Cipta © 2010, BB-Biogen<br />growing the callus on MS medium containing 2,4-D 3 mg/l +<br />thidiazuron 0.5 mg/ l. Formation of callus that can regenerate<br />into shoots should use an MS medium containing 2,4-D 3<br />mg/l + thidiazuron 0.5 mg/ l followed by subculture on MS<br />medium + BAP 3 mg/l + thidiazuron 0.5 mg/l to induce<br />shoot elongation. Medium ½ MS and 1 MS containing NAA<br />0.5-1.0 mg/l can be used to induce root formations on shoot<br />culture of chili cv Gelora but not for cv Chili 109.</p>


HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.


2017 ◽  
Vol 10 (1) ◽  
pp. 35-45
Author(s):  
N.F. Lunkova ◽  
N.A. Burmistrova ◽  
M.S. Krasavina

Background:A growing part of the root is one of the most active sinks for sucrose coming from source leaves through the phloem. In the root, sucrose is unloaded from conducting bundles and is distributed among the surrounding cells. To be involved in the metabolism, sucrose should disintegrate into hexoses by means of degrading enzymes.Aims:The aim of this research was to explore the possibility of the involvement of one such enzymes, invertase, in phloem unloading as well as distribution of its activity in the functionally different tissues of the plant root tips.Method:To estimate the enzyme activities in root tissues, we applied two techniques: the histochemical method using nitro blue tetrazolium. The localization of phloem unloading was studied with carboxyfluorescein, a fluorescent marker for symplastic transport.Results:Invertase activity was not detected in the apical part of the meristem. It appeared only between the basal part of this zone and the beginning of the elongation zone. There is the root phloem unloading in that area. Invertase activity increased with increasing the distance from the root tip and reached the highest values in the region of cell transition to elongation and in the elongation zone. The activities of the enzyme varied in different tissues of the same zone and sometimes in the neighboring cells of the same tissue. Biochemical determination of invertase activity was made in the maize root segments coincident to the zones of meristem, cell elongation and differentiation. The results of both methods of determination of invertase activity were in agreement.Conclusion:It was concluded that phloem unloading correlated with invertase activity, possibly because of the activation of invertase by unloaded sucrose. Invertase is one of the factors involved in the processes preparing the cells for their transition to elongation because the concentration of osmotically active hexoses increases after cleavage of sucrose, that stimulates water entry into the cells, which is necessary for elongation growth.


1990 ◽  
Vol 45 (6) ◽  
pp. 602-606 ◽  
Author(s):  
B. Merkel ◽  
J. Reichling

Abstract Unorganized callus and leaf/root-differentiating callus cultures of Pimpinella major have been established in liquid nutrient medium. Their capacity to accumulate rare phenylpropanoids such as epoxy-pseudoisoeugenol tiglate, epoxy-anol tiglate and anol tiglate was compared with that of seedlings and whole plants. The unorganized callus cultures were not able to accumulate any phenylpropanoids. In comparison, the leaf/root-differentiating callus culture promoted the accumulation of epoxy-pseudoisoeugenol tiglate (up to 90 mg/100 g fr.wt.) but not that of anol-derivatives. The accumulated amount of EPT in PMD-SH was comparable with that in plant seedlings.


Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1102
Author(s):  
Galina N. Raldugina ◽  
Sergey V. Evsukov ◽  
Liliya R. Bogoutdinova ◽  
Alexander A. Gulevich ◽  
Ekaterina N. Baranova

In this study the transgenic lines (TLs) of tobacco (Nicotianatabacum L.), which overexpress the heterologous gene encoding the bacterial enzyme choline oxidase were evaluated. The goal of our work is to study the effect of choline oxidase gene expression on the sensitivity of plant tissues to the action of NaCl. The regenerative capacity, rhizogenesis, the amount of photosynthetic pigments and osmotically active compounds (proline and glycine betaine) were assessed by in vitro cell culture methods using biochemical and morphological parameters. Transgenic lines with confirmed expression were characterized by high regeneration capacity from callus in the presence of 200 mmol NaCl, partial retention of viability at 400 mmol NaCl. These data correlated with the implicit response of regenerants and whole plants to the harmful effects of salinity. They turned out to be less sensitive to the presence of 200 mmol NaCl in the cultivation medium, in contrast to the WT plants.


Biomedicines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 851
Author(s):  
Svetlana I. Dorovskikh ◽  
Evgeniia S. Vikulova ◽  
Elena V. Chepeleva ◽  
Maria B. Vasilieva ◽  
Dmitriy A. Nasimov ◽  
...  

This work is aimed at developing the modification of the surface of medical implants with film materials based on noble metals in order to improve their biological characteristics. Gas-phase transportation methods were proposed to obtain such materials. To determine the effect of the material of the bottom layer of heterometallic structures, Ir, Pt, and PtIr coatings with a thickness of 1.4–1.5 μm were deposited by metal–organic chemical vapor deposition (MOCVD) on Ti6Al4V alloy discs. Two types of antibacterial components, namely, gold nanoparticles (AuNPs) and discontinuous Ag coatings, were deposited on the surface of these coatings. AuNPs (11–14 nm) were deposited by a pulsed MOCVD method, while Ag films (35–40 nm in thickness) were obtained by physical vapor deposition (PVD). The cytotoxic (24 h and 48 h, toward peripheral blood mononuclear cells (PBMCs)) and antibacterial (24 h) properties of monophase (Ag, Ir, Pt, and PtIr) and heterophase (Ag/Pt, Ag/Ir, Ag/PtIr, Au/Pt, Au/Ir, and Au/PtIr) film materials deposited on Ti-alloy samples were studied in vitro and compared with those of uncoated Ti-alloy samples. Studies of the cytokine production by PBMCs in response to incubation of the samples for 24 and 48 h and histological studies at 1 and 3 months after subcutaneous implantation in rats were also performed. Despite the comparable thickness of the fibrous capsule after 3 months, a faster completion of the active phase of encapsulation was observed for the coated implants compared to the Ti alloy analogs. For the Ag-containing samples, growth inhibition of S. epidermidis, S. aureus, Str. pyogenes, P. aeruginosa, and Ent. faecium was observed.


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