scholarly journals Effects of silver nitrate (AgNO3) on growth and anatomical structure of vegetative organs of liquorice (Glycyrrhiza glabra L.) under in vitro condition

Plant Omics ◽  
2018 ◽  
pp. 153-160
Author(s):  
Farnaz Tahoori ◽  
Ahamd Majd ◽  
Taher Nejadsattari ◽  
Hamideh Ofoghi ◽  
Alireza Iranbakhsh

Liquorice (Glycyrrhiza glabra L.) has been used worldwide as a medicine for a long time. In this research, the effect of silver nitrate (AgNO3) as a growth regulator and anti-ethylene in in vitro culture was investigated on growth and anatomical structure of vegetative organs (root, hypocotyl, shoot, leaf) as well as the number of stomata and trichomes in the leaves of liquorice under in vitro culture condition. The seeds were cultured in MS culture media containing different concentrations of AgNO3 (0, 2, 4, 8, and 10 mg L-1). Investigations on 20-day seedlings after three replications showed a significant increase in length and growth of roots, hypocotyls and shoots, and decreased number of stomata and trichomes in the samples treated with AgNO3 (P≤0.05). The effects of AgNO3 on anatomical structures of the organs included the increased cell division in root and shoot tips, reduced vascular tissues and sclerenchyma-fiber (with lignified cell walls), increased thickness of Casparian strip and cell walls of endodermis, reduced thickness of epidermis and increased intercellular spaces in mesophyll. The leaf area was measured in the 4-month plantlets, showing a significant increase in the samples treated with AgNO3. Furthermore, there was significant difference in increased leaf area applying 10 mg L-1 treatment and other concentrations as well as between the concentrations of 2 and 8 mg L-1. It seems that these results are due to the inhibitory effects of AgNO3 on the production and function of ethylene and the plant strategy to increase the tolerance against silver metal.

2016 ◽  
Vol 5 (2) ◽  
pp. 140-146 ◽  
Author(s):  
Aiyatullah Shah ◽  
Muzafar Ahmad Rather ◽  
Aabid Manzoor Shah ◽  
Saleem Mushtaq ◽  
Aehtesham Hussain ◽  
...  

1992 ◽  
Vol 6 (1) ◽  
pp. 13-15
Author(s):  
Пенка Робева-Давидова ◽  
Веселка Гюлева ◽  
Атанас Атанасов ◽  
П. Робева-Давидова ◽  
В. Гюлева ◽  
...  

2008 ◽  
Vol 20 (1) ◽  
pp. 106
Author(s):  
M. J. Sansinena ◽  
D. Owiny ◽  
R. S. Denniston ◽  
D. Salamone ◽  
D. Barry

The riverine rabbit (Bunolagus monticulares), one of South Africa's most threatened mammals, with an estimated population size under 250, was upgraded from endangered to critically endangered in 2002. The low number of riverine rabbits precludes any attempts of nuclear transfer (NT) using intraspecific oocytes; therefore, the overall aim of this study was to assess the ability of the domestic rabbit (Oryctolagus cuniculus) oocyte to reprogram the somatic cell of the endangered riverine rabbit by interspecies NT. A preliminary study evaluated the effect of timing of enucleation after induction of ovulation (h post-hCG). A second study assessed the effects of two activation protocols. In addition, since the unique characteristics of the rabbit zona pellucida affect the speed of micromanipulation, different exposure periods to UV light at enucleation were evaluated. Adult domestic Californian rabbits were treated with eCG for 72 h, and ovulation was induced by hCG administration. Oocytes were collected by retrograde flushing at 12–14 h or 16–18 h post-hCG administration and stripped of cumulus investments with 0.5% hyaluronidase in Ca-Mg-free PBS. Metaphase-II oocytes were selected by visualizing the first polar body. Oocytes were stained with 2 mg mL–1 Hoechst 33342 for 5 min, and metaphase plates were removed with a 25–30 μm (O.D.) borosilicate beveled, spiked pipette after exposure to <5 or 30–40 s of UV light. Adult adipose-derived riverine rabbit fibroblasts grown to confluency in DMEM with 10% FCS were used as donor cells and fused with 2 consecutive DC pulses (3.2 kV cm–1, 45 μs). After reconstruction, couplets were randomly assigned for activation by either a second set of electrical pulses or incubation with ionomycin, followed by 1 h of incubation in 2 mm 6-DMAP. Embryos were co-cultured with a bovine oviductal cell monolayer in DMEM with 10% FCS and assessed for cleavage after 36 h of in vitro culture. There was a significant difference in the number of cleaved embryos from oocytes collected at 12–14 h post-hCG (n = 50) or 16–18 h post-hCG (n = 51) administration (57% v. 0% cleaved; P < 0.05). No significant difference was detected in embryos developing after electrofusion v. ionomycin activation treatments. However, a significantly greater number (P < 0.05) of embryos cleaved from oocytes exposed to <5 s UV than from oocytes exposed to 30–40 s UV (Table 1). A total of 20 embryos (4-cell to 16-cell stages) were surgically transferred to the oviducts of 4 adult New Zealand white synchronized recipients after 48 h of in vitro culture. Two recipients (<5 s UV exposure treatment group) were diagnosed pregnant by abdominal palpation at 15 days post-transfer; pregnancies were subsequently lost by Day 30, with placental tissues recovered. This preliminary study indicates the domestic rabbit oocyte is capable of reprogramming riverine rabbit donor cells. In addition, the time of oocyte collection after ovulation induction and the UV exposure period during enucleation have an effect on the efficiency of interspecies NT and embryo development in this species. Table 1. Effect of UV exposure during enucleation on the in vitro development of interspecies nuclear transfer riverine rabbit embryos


2013 ◽  
Vol 25 (1) ◽  
pp. 219
Author(s):  
J. H. Moon ◽  
S. J. Kim ◽  
J. T. Kang ◽  
S. J. Park ◽  
J. Y. Choi ◽  
...  

Seminal plasma consisting of carbohydrates, proteins, and lipids not only serves as a nutritive and protective medium for sperm cells but also play a pivotal role in inducing the tolerance to pre-existing immune cells as well as improving the intra-uterine conditions for implantation of fertilized embryos (Guerin et al. 2009 Hum. Reprod. Update 15, 517–535). However, the effects of seminal plasma in in vitro culture of fertilized embryos are unknown. In the present study, the seminal plasma was separated from the second fraction of a normal farm boar (n = 1) by centrifugation and filtered seminal plasma was stored at –30°C until use. In a preliminary experiment, the optimal activity of seminal plasma was evaluated by incubating the embryos for different time intervals. To investigate the developmental rates, electrically (EA) (triplicates, n = 490) or chemically (CA) (quintuplicates, n = 599) activated 2-day-old porcine embryos were incubated for 3 h in PZM-5 medium (Funakoshi Co., Tokyo, Japan, Catalog no. IFP0410P) containing 0% (EA: n = 122 and CA: n = 152), 0.1% (EA: n = 123 and CA: n = 148), 0.5% (EA: n = 122 and CA: n = 150), or 1% (EA: n = 123 and CA: n = 149) seminal plasma. Similarly, the developmental rate of chemically activated 2-day-old somatic cell nuclear transferred porcine embryos (quadruplicates, n = 239) was studied after incubation with 0% (n = 119) or 0.1% (n = 120) seminal plasma for 3 h. A significant difference was noticed only in the rate of blastocyst formation in the chemically activated embryos treated with 0.1% seminal plasma (31.7 v. 24.8% in the 0% group, ANOVA; P < 0.05; Prism5, GraphPad Software Inc., La Jolla, CA, USA). None of the treatments showed a significant effect on the cleavage rate and cell numbers of blastocysts. In conclusion, the seminal plasma did not show any harmful effect on early embryos development. Furthermore, the seminal plasma (0.1%) improved the rate of blastocyst formation among the chemically activated nuclear transferred embryos. The results of this preliminary study suggest that the addition of seminal plasma during embryo transfer could increase the rate of pregnancy in pig. This study was supported by MKE (#10033839-2012-21), IPET (#311011-05-1-SB010), the Research Institute for Veterinary Science, and TS Corporation.


2017 ◽  
Vol 29 (1) ◽  
pp. 188
Author(s):  
N. C. Negota ◽  
L. P. Nethenzheni ◽  
M. L. Mphaphathi ◽  
D. M. Barry ◽  
T. L. Nedambale

The in vitro culture media and assisted hatching techniques remain challenging obstacles to be utilised widely. Mechanical, chemical, enzymatic thinning, and laser-assisted techniques have been used previously but information is still lacking on its application in livestock. The aim of this study was to compare the effect of 2 in vitro culture media (Hamster F10 and TMC-199) and 4 (mechanical, chemical, enzymatic, and laser) assisted hatching techniques on blastocyst formation and hatching rate using murine embryos as a model. The C57/b and Balb/c breeds were raised until they reached maturity and bred naturally to produce F1 generation. The light in the breeding house was controlled at 14 h light and 10 h dark. Feed and water were provided ad libitum for the mice. Superovulation of females were stimulated using equine chorionic gonadotropin and human chorionic gonadotropin. The F1 generation was used for the collection of the 400 blastocysts and randomly allocated into 4 assisted hatching techniques. Blastocysts were paired into a group of 10 and replicated 4 times for each assisted hatching technique. The general linear model of SAS version 9.4 (SAS Institute Inc., Cary, NC, USA) was used to analyse the data. Assisted hatching techniques of laser, mechanical, enzymatic, and chemical yielded 46.9 ± 37.1, 51.1 ± 40.2, 39.1 ± 35.8, and 33.3 ± 4.5%, respectively, under in vitro culture of Hamster F10. The TCM-199, laser, mechanical, enzymatic, and chemical assisted hatching techniques yielded 56.3 ± 43.3, 52.6 ± 35.5, 49.2 ± 37.5, and 33.9 ± 35.5%, respectively, with a significant difference. There was no significant difference observed in assisted hatching techniques and Hamster F10 culture medium. However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM than cultured in Hamster F10. Hatching rate of blastocysts increased from chemical, enzymatic, mechanical, and laser with response to Hamster F10 and TCM; thus, laser is a suitable assisted hatching technique with TCM-199.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 330-331
Author(s):  
Yu Liu ◽  
An Gang Lou ◽  
Shuo Yang ◽  
Zhong Shu Li ◽  
Nan-Zhu Fang

Abstract The risk of developmental block in mammal’s embryos is high during in vitro as compare to in vivo environment because the in vitro embryo-culture systems are suboptimal. During in vitro-culture the balance between ROS production and elimination is disturbed and may lead to 2-cell block in mouse embryos [1]. In the current study, we investigated the effects of Glu as anti-developmental block during IVC on ZGA and MZT on mouse embryos. The mouse embryos were divided into control and different level of Glu treated group. The cleavage rate was determined, the ROS and GSH level was investigated using DCHF-DA and CMF2HC respectively. The mRNA expression level of ZGA marker gene such as Eif-1α, Muerv l, Zscan4d and Hsp70.1 was analyzed among the groups using RT-PCR. The transition rate from 2-cell to 4-cell was significantly higher in 6mmol/L Glu treated group as compare to control and others treated groups. No significant difference was recorded in the level of ROS and GSH during MZT stage among the different groups. The mRNA expression level of ZGA marker gene was significantly increased at middle and late stage in 6mmol/L Glu treated group as compare to control and others treated groups. In conclusion, this study shows that the concentration of 6mmol/L Glu could maintain the dynamic balance of GSH and ROS, increase the expression of ZGA marker gene and maintain its high expression pattern of time series, directly participate in the ZGA activated process; ultimately reduce the risk of developmental block to ensure the successful completion of MZT. Reference [1] Lee MT, Bonneau AR, Giraldez AJ.Zygotic Genome Activation during the Maternal-to-Zygotic Transition. Annual Rev Cell Dev Biol [J], 2014, 30:581–613.


2004 ◽  
Vol 16 (2) ◽  
pp. 195
Author(s):  
Y.H. Choi ◽  
D.D. Varner ◽  
K. Hinrichs

Research on in vitro culture of equine embryos has been scant, due to failure of equine in vitro fertilization to be repeatably successful. We have recently obtained high fertilization rates of equine oocytes via intracytoplasmic sperm injection (ICSI) using a piezo drill (Choi et al., 2002 Reproduction 123, 455–465). Culture of presumptive zygotes in G1.2/2.2 medium resulted in 63% cleavage and an average of 15 cells at 4d, but only 2 to 9% blastocyst development at 7 days (Choi et al., 2003 Theriogenology 59, 1219–1229). In the present study, we evaluated the effect of two different culture media, G1.3/G2.3 v. DMEM/F-12, with or without FBS, on blastocyst development after ICSI. Oocytes were collected from slaughterhouse-derived ovaries by follicular scraping and were matured in vitro for 24h in M199 with 10% FBS and 5μUmL−1 FSH. After culture, oocytes having a polar body (198/305; 65%) were fertilized by ICSI with frozen-thawed equine sperm using a piezo drill. Presumptive zygotes were cultured in 1 of 4 media: G1.3/G2.3 (which includes 0.8% BSA) with or without 10% FBS, or in DMEM/F-12 with 0.5% BSA, with or without 10% FBS. Culture was performed in microdroplets at 5μL/zygote under oil at 38.2°C in an atmosphere of 5% CO2, 5% O2 and 90% N2 for 7.5 days. In G1.3/2.3 treatments, G1.3 media were completely refreshed at 48h, zygotes were transferred to G2.3 (with or without FBS as per the first stage) at 96h, and were completely refreshed with the same media at 144h. In DMEM/F-12 treatments, media were completely refreshed every other day. Three to 5 replicates were performed in each treatment, and data were analyzed by chi-square test. There were no significant differences in cleavage rates (59–64%) among treatments. The rate of development to blastocyst, per oocyte injected, in G1.3/G2.3/BSA (1/49, 2%) was significantly lower (P&lt;0.05) than that for the other three treatments: G1.3/2.3/BSA/FBS (9/49, 18%), DMEM/F-12/BSA (9/50, 18%), or DMEM/F-12/BSA/FBS (10/50, 20%). There was no significant difference in blastocyst development among the latter three treatments. These findings indicate that G1.3/2.3 media with BSA only do not adequately support growth of equine embryos. Development of up to 20% of injected oocytes to the blastocyst stage in G media supplemented with FBS, in DMEM/F-12/BSA or in DMEM/F-12/BSA/FBS represents the highest in vitro equine blastocyst rate in medium alone (i.e. without co-culture) yet reported. The success of DMEM/F-12 as an embryo culture medium may provide a relatively simple basis for equine in vitro culture programs. To determine whether this medium was able to support further developmental competence, we cultured equine embryos resulting from nuclear transfer of in vitro-matured oocytes in DMEM/F-12+10% FBS (without BSA). We transferred 4 resulting blastocysts to recipient mares by transcervical transfer; one pregnancy is ongoing at 230d gestation at the time of this writing. This work was supported by the Link Equine Research Endowment Fund, Texas A&amp;M University.


2021 ◽  
Vol 89-90 ◽  
pp. 89-100
Author(s):  
Iryna Kovalyshyn ◽  
Andrii Pinchuk ◽  
Artur Likhanov

Quantitative morpho-anatomical features of leaves of nine Clematis taxa (C. alpina ‘Pamela Jackman’, C. macropetala ‘Maidwell Hall’, C. integrifolia ‘Aljonushka’, C. ispahanica ‘Zvezdograd’, C. fargesii ‘Paul Farges’, C. texensis ‘Princess Diana’, C. tibetana, C. viticella, and C. heracleifolia) were determined with the aim to analyze their adaptation to the environmental conditions.Among investigated clematises, there were plants with hypostomatic (C. viticella, C. fargesii ‘Paul Farges’, C. heracleifolia, C. texensis ‘Princess Diana’, C. macropetala ‘Maidwell Hall’, and C. alpina ‘Pamela Jackman’), and amphistomatic leaves (C. ispahanica ‘Zvezdograd’ and C. tibetana). In C. integrifolia ‘Aljonushka’ leaves were hypostomatic, but few solitary stomata were also present on the adaxial surface. In the leaves of investigated taxa, the palisade coefficient ranged from 27.3% (C. alpina ‘Pamela Jackman’) to 49.9% (C. tibetana). The leaves also differed significantly in size. In particular, leaves of C. integrifolia ‘Aljonushka’ were almost ten times smaller than such of C. heracleifolia.As a result of UPGMA clustering, the plants that can survive in severe windy weather in open rocky areas, Clematis tibetana and C. ispahanica ‘Zvezdograd’, were joined in a separate cluster. The second cluster combined C. alpina ‘Pamela Jackman’ and C. macropetala ‘Maidwell Hall’ – cultivars blooming in the spring, during a period of significant difference in daily temperatures. A relatively small leaf area in plants from these two clusters may indicate an adaptation by reducing the transpiration area and general windage. The third cluster united the rest of investigated taxa, mostly – the mesophytic plants with a relatively large leaf area. However, due to similar morpho-anatomical structure of the leaf, the third cluster also comprised C. integrifolia ‘Aljonushka’ with the smallest leaves.


Data in Brief ◽  
2021 ◽  
pp. 107535
Author(s):  
Gayathree Karthikkeyan ◽  
Ashwini Prabhu ◽  
Ravishankar Pervaje ◽  
Sameera Krishna Pervaje ◽  
Prashant Kumar Modi ◽  
...  

2014 ◽  
Vol 6 (1) ◽  
pp. 82-84
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Sunitibala Devi HUIDROM

Silver nitrate (AgNO3) was used under in vitro conditions to study the response of ginger cultivars ‘Nadia’ and ‘Baishey’ under ex vitro. Micropropagated plants treated with AgNO3 showed significant difference (p


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