In situ seed development and in vitro regeneration of three difficult-to-propagate Lepidosperma species (Cyperaceae)

2010 ◽  
Vol 58 (2) ◽  
pp. 107 ◽  
Author(s):  
Andrea Kodym ◽  
Shane Turner ◽  
John Delpratt
Keyword(s):  
In Vitro Culture ◽  
In Vitro Regeneration ◽  
Field Studies ◽  
Fruit Production ◽  
Germination Response ◽  
Immature Seeds ◽  
Short Period ◽  
Viable Seeds

Field studies of fruit production from Lepidosperma concavum R.Br., L. laterale R.Br. and L. longitudinale Labill. showed that large proportions (21–77%) of fruits were unfilled and that filled and unfilled fruits looked alike. Bagging of inflorescences demonstrated that filled fruits tended to be shed, while empty fruits remained within the inflorescence. Time of collection was critical for obtaining viable seeds, with successful harvesting limited to a short period (weeks) after maturation. The timing of flowering and fruit maturation were fairly consistent between species, populations and years in our study area. In L. concavum fruit production was increased in cultivation compared with wild populations. In all three species, very little or no germination of fruits occurred under nursery conditions. In vitro culture initiation was attempted using intact fruits, nicked fruits and seeds on 1/2MS (Murashige and Skoog) medium with 1 µM zeatin and 0.5 µM gibberellic acid in darkness. Culture of intact fruit resulted in no germination, while nicked fruit showed some germination response. Best results were achieved from seeds with germination occurring as early as 7 to 18 days depending on the species. Germination of L. concavum, L. laterale and L. longitudinale was 86%, 64% and 83% respectively within 5 weeks. Germination response was strongly influenced by seed maturity. Mature seeds germinated significantly faster than immature seeds. On a small proportion of cultured seeds, calli formed and differentiated into numerous plantlets on growth regulator-free medium. Given the promising results observed in this study, in vitro culture appears to be a practical means of mass propagating Lepidosperma species.

scholarly journals Therapeutical aspect of trichomoniasis

2003 ◽  
Vol 131 (3-4) ◽  
pp. 156-162 ◽  
Author(s):  
Jelica Vukicevic ◽  
Jasmina Jankicevic
Keyword(s):  
Trichomonas Vaginalis ◽  
Communicable Disease ◽  
Repeated Administration ◽  
Transmitted Infection ◽  
Microbiological Test ◽  
Clinical Forms ◽  
Short Period ◽  
Significant Difference

Trichomoniasis is frequent, parasitic and sexually transmitted infection of genitourinary tract. It is treated by metronidazole (5-nitroimidazole) according to protocol recommended by Center for Disease Control (CDC formerly called: Communicable Disease Center) [19]. The resistance of Trichomonas vaginalis (TV) strains to metronidazole (MND) was described in USA in 1960, and later on in many European countries [8, 9, 10, 11, 12, 13]. In these cases, due to persistent trichomonas infection, it is necessary to repeat MND treatment with moderate modification of dose and/or length of its application. Nevertheless, oncogenic and toxic effects of MND have to be taken into consideration. OBJECT The aim of this study was to investigate and analyze the incidence of TV in STD and lower susceptibility of certain TV strains to MND were analyzed. MATERIAL AND METHODS In three-year period (1999-2001) 612 patients (244 females and 368 males) suspected of STD were examined clinically and microbiologically at the Institute of Dermatovenereology in Belgrade. The patients detected for TV were treated according to CDC protocol. The affected were considered cured if there was no manifest clinical infection, and no TV verified by microbiological test. Results TV was isolated in 216 patients (35.29 % of all subjects). Trichomonas infection was found in 90 (36.88 %) out of 244 tested females and in 126 (32.34 %) of 368 males. Clinically manifested infection, with extensive urethral and vaginal secretion, was recorded in 161 patients, while the asymptomatic form was found in 55 subjects. This result indicates the predominance of manifested trichomonas infections (75.54 % of cases). The difference of distribution of clinical forms of trichomoniasis, in relation to sex, was not statistically significant (c2=0.854; p>0.05). The patients with verified trichomonas infection were treated by metronidazole according to CDC protocol. The recommended therapeutical scheme consisted of three phases proceeding in succession, in so far TV had not been eliminated by previous one. The number of cured patients, according to therapeutical phases, was shown in Table 4. Three patients (1.39 %, 2 males and 1 female) were not cured in spite of all three completed phases of therapeutical protocol. In all three cases, TV was eliminated by MND application in dose of 3 g/daily, during two days. The failure of minute MND treatment was analyzed in relation to clinical forms of the infection (manifested or asymptomatic), as well as in relation to types of infection (single- or associated infection). The incidence of refractory trichomoniasis treated by a single metronidazole dose of 2 g was significantly higher in the group of patients with polyinfection (c2=18.270 p<0.01). There was no significant difference of resistance to a single MND dose between the groups with manifested and asymptomatic trichomoniasis (c2=0.321; p<0.01). The prevalence of TV in vaginal and urethral smears indicates the significant incidence of trichomoniasis in STD. TV was more frequently isolated in patients with clinically manifested infection. TV susceptibility to MND was tested in vitro in aerobic and anaerobic conditions. The resistance of strains under in vitro conditions did not correlate with refractory feature of trichomoniasis to MND application [7 17, 18]. The success of trichomoniasis treatment depends upon multiple factors, including: a) TV susceptibility to drug, b) intravaginal redox potential, c) drug concentration in situ, d) associated microorganisms that may modify the amount of the drug available in situ [7, 18, 21]. The results of our investigation argue for the latter item, verifying that TV resistance to MND is higher in patients with polyinfection in relation to those with monoinfection (significant difference, c2=18.270; p<0.01). Repeated administration of low metronidazole doses may prolong the therapy of trichomonas infections, while application of high doses (over 3 g/day) may result in undesired complications. Given the well-known fact that repeated sublethal doses induce the resistance, would it be more beneficial to begin with slightly higher metronidazole dose (3 g/day) during short period of time (3-5 days)? This will be the subject of our further investigation.

Regeneration of peanut (Arachis hypogaea) plantlets by in vitro culture of immature leaves

1981 ◽  
Vol 59 (5) ◽  
pp. 826-830 ◽  
Author(s):  
L. A. Mroginski ◽  
K. K. Kartha ◽  
J. P. Shyluk
Keyword(s):  
In Vitro Culture ◽  
Arachis Hypogaea ◽  
Environmental Conditions ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Bud Regeneration ◽  
Arachis Hypogaea L ◽  
Murashige And Skoog Medium ◽  
Germinated Seeds

The in vitro regeneration of buds, shoots, and roots from immature leaves of 3- to 5-day-old peanut (Arachis hypogaea L. cv. Colorado Manfredi) seedlings was studied under defined nutritional, hormonal, and environmental conditions. The first two leaves (2–5 mm in length) removed from aseptically germinated seeds were cultured on Murashige and Skoog medium containing vitamins as in B5 medium and 0.8% agar, supplemented with 12 combinations of naphthaleneacetic acid (NAA) (0.01 to 4 mg/L) and benzyladenine (BA) (1 and 3 mg/L). Bud regeneration occurred in all hormone combinations, but the maximum number of buds was regenerated at a concentration of 1 mg/L each of NAA and BA. Although bud regeneration was maximum with 2- to 5-mm-long leaflets, some success was also obtained with leaflets 8–13 mm long. However, no buds were regenerated when fully expanded leaflets were cultured.Development of buds into shoots was readily achieved by transferring regenerated buds into fresh medium containing 0.01 mg/L NAA and 1 mg/L BA. A few roots were induced to grow when callus with buds was also transferred to medium devoid of hormones. So far, bud regeneration from immature leaves has been induced in vitro in 5 of the 10 cultivars tested.

Passage-Dependent Cancerous Transformation Of Human Mesenchymal Stem Cell Of Adipose Origin During In Vitro Culture

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4862-4862
Author(s):  
Jung Ah Kim ◽  
Qute Choi ◽  
Kyong Ok Im ◽  
Ji Seok Kwon ◽  
Si Nae Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
In Vitro Culture ◽  
Telomere Length ◽  
Conflicts Of Interest ◽  
Bac Clone ◽  
Interphase Cell ◽  
Cytogenetic Changes ◽  
Chromosomal Changes

Abstract Introduction In vitro culture of adult human mesenchymal stem cells (hMSCs) can induce cancerous transformation, depending on environmental factors. To evaluate the passage dependent chromosomal changes of hMSCs toward malignant transformation, we passaged adipose origin hMSC to 9th passage and analyzed cytogenetic change, molecular cytogenetic changes, and telomere length variations on every passage. Methods On each passage, in situ karyotyping was performed on 3 separate batches with subsequent Giemsa staining. Karyotyping was analyzed using Metafer system (MetaSystems, Altlussheim, Germany). For analysis of nonproliferating interphase cell, each chromosome were counted with centromere fluorescent in situ hybridization (FISH) using Same Day OligoFISH™(Cellay Inc., Cambridge, Massachusetts, USA). Telomere length was analyzed using FISH technique with a Cy3-lableled Telomere peptide nucleic acid (PNA) FISH kit (DakoCytomation Denmark A/S, Glostrup, Denmark). To confirm the chromosomal translocation appeared by in situ karyotyping, we made home-brew FISH probe with bacterial artificial chromosome(BAC) clone and quantitated the proportion of abnormal cells by interphase FISH. Results On 5th passage, translocation and polysomy of chromosome 7 and 9 appeared, and on 6th passage, additional translocation t(6; 10) appeared. ISCN Karyotypes of chromosomal changes from 5th passage to 7th passage were 48,XX,+7,t(7;22)(q11.22;q13.3),+9[4]/46,XX[21] → 47,XX,+7[2]/47,XX,t(6;10)(q21;q25.1),+7[2]/46,XX[13] → 48,XX,+7,t(7;22)(q11.22;q13.3),+9[6]/ 47,XX,+7[5]/46,XX[9]. Telomere length was decreased on late passages. Fusion signal of t(7;22) on passage 5(fig 1) and that of t(6;10) on passage 6(fig 2) were confirmed by BAC clone. Conclusions The behavior of late passage (from passage 5) follows a cytogenetic profile similar to that of transformed cancer cells. Cytogenetic abnormalities which were not observed in earlier passage, showed up and disappeared, but eventually persisted during passages. We suggest in vitro environment cause hMSCs to undergo cancer-like cytogenetic changes. It is of great importance to test safeguards for clinical applications of human stem cells manufactured in vitro. Disclosures: No relevant conflicts of interest to declare.

Fertilization biology of the abalone Haliotis laevigata: laboratory and field studies

1999 ◽  
Vol 56 (9) ◽  
pp. 1668-1678 ◽  
Author(s):  
Russ Babcock ◽  
John Keesing
Keyword(s):  
Field Experiments ◽  
Nearest Neighbor ◽  
Field Studies ◽  
Fertilization Success ◽  
Limiting Factor ◽  
Fertilization Rates ◽  
Abalone Species ◽  
Nearest Neighbor Distances

A combination of laboratory and field experiments with the commercial abalone species Haliotis laevigata showed that fertilization may be a limiting factor in some exploited populations when distances separating spawning individuals are too large. The effects of gamete age, gamete concentration, and gamete contact time in the laboratory were used to model fertilization success in situ and compared with experimental fertilization rates in the field. Highest fertilization rates in vitro (80%) were found for sperm concentrations in the range of 1 × 104 to 1 × 106·mL-1. Fertilization rates of 48 ± 1.7% (95% CI) were measured at separation distances of 2 m and dropped to 2.8 ± 0.7% (95% CI) at 16 m downstream, agreeing closely with rates predicted by the model. Recruitment failures reported for South Australian populations of H. laevigata have occurred when densities fell below ca.0.3 animals·m-2, or mean nearest-neighbor distances between 1 and 2 m. This density corresponds well to critical nearest-neighbor distances for fertilization success. Stocks at higher densities are predicted to have higher fertilization rates (ca.90%) such that fertilization success is not a factor limiting recruitment.

Foetal Human Melanocytes: In Situ Detection, In Vitro Culture and Differentiation Characteristics at 6–11 Weeks EGA

1996 ◽  
Vol 9 (3) ◽  
pp. 126-133 ◽  
Author(s):  
I. CAROLINE LE POOLE ◽  
RENE M.J.G.J. VAN DEN WIJNGAARD ◽  
RONALD P. VERKRUISEN ◽  
WOUT H. LAMERS ◽  
DIRK TROOST ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Human Melanocytes ◽  
In Situ Detection

scholarly journals In-vitro Regeneration from Direct and Indirect Organogenesis of Crescentia alata Kunth an Important Multipurpose Medicinal Tree

2021 ◽  
pp. 48-58
Author(s):  
R. Abinaya
Keyword(s):  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Direct Organogenesis ◽  
Ms Medium ◽  
Indirect Organogenesis ◽  
Medicinal Tree ◽  
Short Period ◽  
In Vitro Clonal Propagation

In this present work, an in-vitro regeneration protocol for Crescentia alata (C. alata) was developed using various explants on Murashige and Skoog (MS) medium augmented with different concentrations and combinations of plant growth regulators (PGRs) for direct and indirect regeneration. The direct organogenesis was established from nodes and internodes on MS medium supplemented with cytokinins and auxins. The indirect organogenesis via callus phase was obtained from leaf, nodes and internodes on MS medium supplemented with different concentrations of PGRs. The high frequency shoot organogenesis were achieved directly from nodal explants were cultured on MS medium supplemented with 3.0 mg/L BAP+0.5 mg/L KIN +1.0 mg/L NAA. Indirect organogenesis callogenic frequency was optimized at the concentration of MS medium containing 1.0 mg/L BAP + 5.0 mg/L IAA. The callus was obtained from all the explants were used, among these explants internodal explants gave best result on MS medium supplemented with different concentrations of cytokinins and auxins for indirect organogenesis experiment. Indirect organogenesis the highest number of shoot regeneration was obtained in MS Basal Medium with 4.0 mg/L BAP + 0.5 mg/L KIN + 2.0 mg/L NAA from internodal explants. For root formation the regenerative shoots which were sub cultured on MS medium containing different ratios of auxins. The rooted plantlets were transferred successfully to the pots containing sterilized soil and were successfully hardened at greenhouse condition for 20 days then exposed to the natural environment. This is the first successful micropropagation report of an efficient and rapid in-vitro clonal propagation protocol for C. alata by direct and indirect shoot organogenesis through various explants, which can be employed for conservation of this important medicinal tree species as well as the utilization of an biologically important active biomolecules. This protocol can be very useful to obtain plants from various explants, without the requirement of meristematic regions, enabling the obtainment of a higher number of plants in short period.

scholarly journals State of populations of rare s pecies Silene jailensis N.I. Rubzov and Heracleum ligusticifolium M. Bieb. on Nikitskaya Yayla

2021 ◽  
Vol 1 (157) ◽  
pp. 114-122
Author(s):  
A. R. Nikiforov
Keyword(s):  
Adverse Effect ◽  
Small Area ◽  
Self Regulation ◽  
Structural Features ◽  
External Threats ◽  
Internal Mechanism ◽  
Viable Seeds ◽  
Destructive Processes

Populations of obligate petrophytes of the Mountain Crimea Heracleum ligusticifolium M. Bieb. (Apiaceae) and Silene jailensis N.I. Rubtzov (Caryophyllaceae) are distinguished by their small number due to the internal mechanism of their self-regulation: in small-area local stony habitats, plants of pregenerative age are regularly eliminated. This mechanis m operates independently of external threats, the adverse effect of which was smoothed out by the structural features of populations: the predominance of long -lived generative plants in S. jailensis, and the abundance of viable seeds in H. ligusticifolium. However, in recent years, the population of the Nikitskaya Yayla has been observed to have a weak lack of seed renewal of Silene jailensis and a complete lack of seed renewal of Heracleum ligusticifolium. This circumstance led to intra-population destructive processes. The probability of degradation and extinction of populations actualized the development of methods of reproduction and maintenance of plants of these species in vitro. By now, there is a stock of plants of these species that can be used for in situ reintroduction.

scholarly journals How Carbon Source and Seedcoat Influence the In Vitro Culture of Peach (Prunus persica L. Batsch) Immature Seeds

HortScience ◽  
2021 ◽  
pp. 1-2
Author(s):  
Margarita Pérez-Jiménez ◽  
Alfonso Guevara-Gázquez ◽  
Antonio Carrillo-Navarro ◽  
José Cos-Terrer
Keyword(s):  
Seed Germination ◽  
Carbon Source ◽  
In Vitro Culture ◽  
Seedling Growth ◽  
Woody Plant ◽  
Prunus Persica ◽  
In Vitro Germination ◽  
Immature Seeds ◽  
Woody Plant Medium

The effects of carbon source and concentration and of seedcoat were tested on the in vitro germination of peach seeds derived from crosses performed in the field. Seeds were extracted from the fruit and cultured in Woody Plant Medium (WPM) supplemented with sucrose, glucose, or sorbitol at concentrations of 15, 30, and 45 g·L−1. The percentage of germination as well as the root and hypocotyl lengths were measured after the stratification process and before acclimatization. Seedcoat did not have any influence on seed germination in any tested media and genotype. Glucose at a concentration of 15 g·L−1 and sucrose at 15, 30, and 45 g·L−1 resulted in greater stem seedling growth. The root developed the most when seeds were cultured in media with 15 or 30 g·L−1 of sucrose.

scholarly journals Composition and Radical Scavenging Activity of the Extracts from Deschampsia Antarctica É. Desv. Plants Grown in Situ and in Vitro

2021 ◽  
Author(s):  
Roman Ivannikov ◽  
Iryna Laguta ◽  
Viktor Anishchenko ◽  
Iryna Skorochod ◽  
Pavlo Kuzema ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Deschampsia Antarctica ◽  
Oh Radicals ◽  
Scavenging Activity ◽  
Hydroxybenzoic Acids

The work was aimed at studying the phytochemicals available in D. antarctica plants from various sites on Antarctic Islands, introducing the plants into in vitro culture and comparing the extracts from the plants grown in situ and in vitro. All the extracts were found to contain high amount of phenols, with luteolin and hydroxybenzoic acids derivatives being the main phytochemicals in the extracts from the plants grown in situ and in vitro. Being diluted by 10 times, the extracts scavenged 50÷90% of DPPH• radicals, 20÷40% of NO• radicals and 40÷60% of OH• radicals. Despite the differences in composition, extracts from the plants grown in situ and in vitro showed similar radical scavenging activity.

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