Basal medium improvement for routine micropropagation of Olea maderensis: physiological comparative studies

2009 ◽  
Vol 39 (4) ◽  
pp. 814-822 ◽  
Author(s):  
Gina Brito ◽  
Conceição Santos
Keyword(s):  
Endangered Species ◽  
Membrane Integrity ◽  
Basal Medium ◽  
Germplasm Preservation ◽  
Leaf Chlorosis ◽  
Chlorophyll A And B ◽  
Leaf Survival ◽  
Mn Concentrations

The routine micropropagation of Olea maderensis (Lowe) Rivas Mart. & Del Arco requires an adequate basal medium. To find the optimal basal medium, shoots were grown on four different media: olive medium (OM) and three modified OM media enriched with 2, 4, and 10 times the iron (Fe), magnesium (Mg), and manganese (Mn) concentrations of the OM medium, respectively (OMG, OMG4, and OMG10). For the elongation–proliferation stage, media were supplemented with 9.12 µmol·L–1 zeatin. Doubling the Fe, Mg, and Mn concentrations (OMG) provided green and healthy shoots, while in other media leaf chlorosis or necrosis and abscission occurred because of either deficiency (OM) or toxicity of the elements (OMG4 and OMG10). Shoots grown in the OMG medium also had the highest elongation–proliferation rates. Physiological studies were then performed only between OMG- and OM-grown shoots. Chlorophyll a and b contents and fluorescence (Fv, Fm) were higher in OMG-grown shoots. Doubling the concentration of Fe, Mg, and Mn (which are important in photosynthesis) stimulated leaf survival and photosynthesis. OMG-grown leaves had higher Mg, Fe, and Mn levels. Compared with field leaves, in vitro leaves had higher protein contents. Other physiological parameters (membrane integrity, water content, and osmolality) did not differ between the two media. The best rooting rates were obtained for shoots grown in OMG (>85%), and plants were successfully acclimatized. These studies improved the efficiency and quality of micropropagation of O. maderensis, thereby allowing germplasm preservation of this endangered species.

In vitro Supplementation of Linoleic Acid Improves Quality of Cryopreserved Buffalo Semen

2020 ◽  
Author(s):  
R Ejaz ◽  
S Qadeer ◽  
M S Ansari ◽  
B A Rakha ◽  
S Shamas ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Membrane Integrity ◽  
Sperm Chromatin ◽  
Standard Procedures ◽  
Buffalo Semen ◽  
Chromatin Integrity ◽  
Live Sperm ◽  
And Control

Present study was designed to evaluate the effect of linoleic acid (LA) supplementation in extender on post thaw quality of cryopreserved buffalo semen. Semen was collected from three adult Nili Ravi buffalo bulls of same age with artificial vagina (42°C) for five weeks (replicates; N=30). Qualified semen ejaculates (>1mL volume, >60% motility, >0.5 billion/mL concentration) were diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0ng mL-1 of LA and were cryopreserved using standard procedures. Sperm motility and plasma membrane integrity were improved plessthan0.05) in extender containing 10.0 ng mL-1 of LA compared to other treatments and control while number of acrosome intact live sperm, chromatin integrity and number of morphologically normal sperms remained the same. In conclusion, LA supplementation in extender at 10.0 ng mL-1 was found to be beneficial to improve post thaw quality of cryopreserved buffalo semen.

scholarly journals Evaluation of sperm quality of Erythrolamprus poecilogyrus sublineatus (Cope, 1860) (Serpentes, Dipsadidae)

2017 ◽  
Vol 77 (3) ◽  
pp. 553-557
Author(s):  
A. C. Silva ◽  
A. S. Varela Junior ◽  
T. F. Cardoso ◽  
E. F. Silva ◽  
D. Loebmann ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Coastal Region ◽  
Rio Grande ◽  
Rio Grande Do Sul ◽  
Sperm Analysis ◽  
Pampa Region

Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

scholarly journals Propagation for the Conservation of Pityopsis ruthii, an Endangered Species from the Southeastern United States

HortScience ◽  
2014 ◽  
Vol 49 (2) ◽  
pp. 194-200 ◽  
Author(s):  
Phillip A. Wadl ◽  
Timothy A. Rinehart ◽  
Adam J. Dattilo ◽  
Mark Pistrang ◽  
Lisa M. Vito ◽  
...  
Keyword(s):  
Endangered Species ◽  
Natural Habitat ◽  
Basal Medium ◽  
Single Individual ◽  
In Vitro Multiplication ◽  
Large Numbers ◽  
Lateral Shoots ◽  
Genetic Swamping ◽  
Germination Techniques

Pityopsis ruthii is an endangered species endemic to the Hiwassee and Ocoee Rivers in Tennessee. As part of a recovery effort focused on P. ruthii, vegetative propagation and in vitro multiplication and seed germination techniques were developed. Plants were vegetatively propagated using greenhouse stock plants and wild-collected stems. Rooting occurred with and without auxin treatments but was greatest when 0.1% indole-3-butyric acid (IBA) talc was applied to the vegetative cuttings; rooting was lowest when flowering stems were used. Pro-Mix BX substrate provided the most consistent rooting. In vitro multiplication was accomplished by the removal of lateral shoots from in vitro-grown plants that were rooted on Murashige and Skoog (MS0) basal medium with 270 clones produced from a single individual after 4 months. Nineteen clones were transplanted and secured with bonded fiber matrix into their natural habitat and 14 survived for 1 year. To avoid genetic swamping of native populations with the introduction of large numbers of genetically identical individuals through clonal propagation, seed-based propagation efforts were explored. Open-pollinated seeds were collected, disinfested and germinated, and seedlings established on MS medium. Seeds were submersed in 70% ethanol for 1 minute and briefly flamed. Seeds were surface-sterilized in a range [10% to 50% (v/v)] Clorox® bleach solutions with vigorous shaking for 20 minutes, rinsed three times in sterile water, and germinated on MS0. Removal of pappus from seeds was required for successful disinfestations, but the bleach concentration was not critical. Successful propagation is a step toward the conservation and recovery of P. ruthii and should allow future reintroduction projects.

scholarly journals Effects of apple and orange juices on quality of refrigerated goat semen

2018 ◽  
Vol 63 (1) ◽  
pp. 53-65
Author(s):  
Ezekiel Adekunle ◽  
James Daramola ◽  
Olusiji Sowande ◽  
John Abiona ◽  
Monsuru Abioja
Keyword(s):  
Sperm Motility ◽  
Orange Juice ◽  
Apple Juice ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Fruit Juices ◽  
Sperm Viability ◽  
In Vitro Storage

This study investigated the effects of apple and orange juices on quality of refrigerated spermatozoa of goat bucks. Semen samples from WAD goat bucks were diluted with Tris-egg yolk extenders each supplemented with apple and orange juices at 0, 2.5, 5, 7.5 and 10/100 ml of diluents. The diluted semen samples were assessed for sperm viability and malondialdehyde (MDA) concentration after in vitro storage for 240 hours at 5oC. The ability to maintain sperm motility was higher in the extenders with 7.5% orange juice followed by 10% apple juice compared to other treatments (P<0.05). The extenders supplemented with 2.5%, 5% and 7.5% apple juice, and 5% orange juice had higher intact acrosome compared to other treatments and the control (P<0.05). The 10% orange juice had higher percentage membrane integrity compared to other treatments. Consistent and reduced (P<0.05) MDA levels were observed in the extenders supplemented with fruit juices and lower MDA was observed in the extenders supplemented with 10% apple juice compared to other treatments and the control (P<0.05). The findings reveal that additions of the fruit juices to semen extenders to maintain the viability of refrigerated spermatozoa were best at concentrations of 10 ml/100 ml of apple juice and 7.5 ml/100 ml of orange juice.

Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants

2016 ◽  
Vol 28 (12) ◽  
pp. 1990 ◽  
Author(s):  
D. Acha ◽  
M. Hidalgo ◽  
I. Ortiz ◽  
M. J. Gálvez ◽  
J. J. Carrasco ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Equus Asinus ◽  
Sperm Membrane ◽  
Freeze Test ◽  
Pooled Samples

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

scholarly journals Effects of four extenders on the quality of frozen semen in Arabian stallions

2019 ◽  
Vol 12 (1) ◽  
pp. 34-40 ◽  
Author(s):  
Mohaammed Saad Alamaary ◽  
Haron Wahid ◽  
Mohamed Ali ◽  
Mark Wen Han Hiew ◽  
Lawan Adamu ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
Before And After ◽  
Semen Preservation ◽  
The Impact

Aim: Different types of extenders have a variety of components which show the tolerance effect on sperm protection during freezing procedures. In the present study, we have examined the impact of the extenders HF-20 and Tris, which were locally manufactured, and they are competing with commercial extenders INRA Freeze® (IMV Technologies, France) and EquiPlus Freeze® (Minitube, Germany) on the quality of horses frozen semen. Materials and Methods: A total of 15 ejaculates from three healthy stallions were collected and cryopreserved in the same environment. Each semen sample collected was divided into four equal parts and processed. All samples were analyzed before and after freezing for motility, viability, plasma membrane integrity, and morphology. Furthermore, twenty mares were inseminated using post-thawed semen. Results: There were no differences observed among all extenders in all the parameters before freezing. Sperm cryopreserved using HF-20 showed better motility, viability, and plasma membrane integrity than Tris extender. The Tris extender showed the most inferior quality of post-thawed semen between all the extenders. HF-20, INRA Freeze®, and EquiPlus Freeze® extenders revealed the same capacity of semen preservation in vitro and in vivo. Conclusion: HF-20 extender has the same quality as INRA Freeze® and EquiPlus Freeze® that can be considered as one of the best extenders for the semen cryopreservation in horses. In contrast, Tris extender needs some degree of improvement.

Cytisus aeolicus Guss. ex Lindl. in vitro cultures and genistin production

2010 ◽  
Vol 5 (1) ◽  
pp. 111-120 ◽  
Author(s):  
Mariella Lucchesini ◽  
Alessandra Bertoli ◽  
Anna Mensuali-Sodi ◽  
Elisa Cappelli ◽  
Cecilia Noccioli ◽  
...  
Keyword(s):  
Casein Hydrolysate ◽  
Basal Medium ◽  
Callus Cultures ◽  
In Vitro Cultures ◽  
Wild Plants ◽  
Potential Production ◽  
Germplasm Preservation ◽  
Dichlorophenoxy Acetic Acid ◽  
Unique Compound

AbstractCytisus aeolicus Guss. ex Lindl. (Fabaceae family, subfamily Faboideae) is an endangered endemic species of the Aeolian Islands, Sicily. In vitro multiplication of C. aeolicus shoots was described in this work and cell cultures were established from cotyledons and hypocotyls to investigate their potential production of isoflavones. Aseptically germinated seeds, cultivated on LS modified basal medium, gave the initial explants used both to induce axillary propagation and callus cultures. The LS (Linsmaier and Skoog) basal medium, supplemented with 0.1 mg L−1 of 6-benzylaminopurine were used to induce axillary propagation. The callus induction was performed using the basal medium added with 5 mg L−1 2,4-dichlorophenoxy acetic acid and 5 mg L−1 kinetin (control medium). Basal medium was also added with 2000 mg L−1 casein hydrolysate (CH) or 900 mg L−1myo-inositol (MI). C. aeolicus callus cultures on CH and MI media produced an unique compound, the isoflavone genistein 7-O-ß-D-glucopyranoside (genistin), which has not previously been isolated from wild plants. Callus cultures grown on the medium containing myo-inositol produced the greatest amount of genistin. C. aeolicus tissue culture procedures could provide suitable plant material both for germplasm preservation (by micropropagation) and for biotechnological selective isoflavone production (by callus culture).

scholarly journals Effect of Addition of Melatonin on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
P. Perumal ◽  
Kezhavituo Vupru ◽  
K. Khate
Keyword(s):  
Membrane Integrity ◽  
Control Group ◽  
Protective Effects ◽  
Sperm Abnormality ◽  
Total Antioxidant ◽  
Biochemical Profiles ◽  
Group 5 ◽  
Group 2

The present study was undertaken to assess the effect of melatonin (MT) on sperm motility, viability, total sperm abnormality, acrosomal and plasma membrane integrity, DNA abnormality, antioxidant profiles such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and total antioxidant capacity (TAC), enzymatic profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT), and biochemical profiles such as malonaldehyde (MDA) production and cholesterol efflux. Total numbers of 30 ejaculates were collected twice a week from eight mithun bulls and semen was split into five equal aliquots, diluted with the TEYC extender. Group 1 has semen without additives (control) and group 2 to group 5 have semen that was diluted with 1 mM, 2 mM, 3 mM, and 4 mM of melatonin, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 0, 6, 12, 24, and 30 h of incubation. Inclusion of melatonin into diluent resulted in significant (P<0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, melatonin at 3 mM has significant improvement in quality of mithun semen than melatonin at 1 mM, 2 mM or 4 mM stored inin vitrofor up to 30 h. It was concluded that the possible protective effects of melatonin on sperm parameters are it prevents MDA production and preserve the antioxidants and intracellular enzymes during preservation.

scholarly journals In Vitro and Cryobiotechnology Approaches to Safeguard Lupinus rivularis Douglas ex Lindl., an Endangered Plant in Canada

Agronomy ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 37
Author(s):  
Elena V. Popova ◽  
Mukund R. Shukla ◽  
Terry McIntosh ◽  
Praveen K. Saxena
Keyword(s):  
Seed Germination ◽  
Endangered Species ◽  
In Situ Conservation ◽  
Germplasm Conservation ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Ex Situ ◽  
Natural Habitats

Conservation of threatened flora in genetic collections ex situ using in vitro culture and cryopreservation is receiving an increasing recognition as a complementary strategy to in situ conservation in natural habitats. The present study is focused on an integrated approach which involves conservation and propagation, emphasizing the usefulness of cryopreservation techniques for germplasm conservation of streambank lupine (Lupinus rivularis Douglas ex Lindl.), an endangered species in Canada. This included in vitro seed germination on Murashige and Skoog basal medium supplemented with 1 µM thidiazuron to induce multiple shoot formation, micropropagation on a medium with 5 µM benzylaminopurine, and in vitro rooting on medium with 20.0 µM indole-3-butyric acid. Cryopreservation of seeds and shoot tips of in vitro grown plants was successful with over 60% seed germination and 62% regrowth of cryopreserved shoot tips, respectively. Plants developed from cryopreserved seeds had chlorophyll contents in leaves and the growth characteristics including the development of inflorescence, similar to plants raised from non-cryopreserved seeds. These results provide further evidence that the combination of micropropagation with cryopreservation of seeds and vegetative parts may effectively facilitate long-term preservation of L. rivularis and other endangered species.

scholarly journals Effects of Cryopreservation on Gene Expression and Post Thaw Sperm Quality of Pacific Abalone, Haliotis discus hannai

2021 ◽  
Vol 8 ◽  
Author(s):  
Shaharior Hossen ◽  
Zahid Parvez Sukhan ◽  
Yusin Cho ◽  
Kang Hee Kho
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Dna Integrity ◽  
Haliotis Discus Hannai ◽  
Pacific Abalone ◽  
Haliotis Discus

Pacific abalone, Haliotis discus hannai, is a high commercial seafood in South-East Asia. The aim of the present study was to determine effects of cryopreservation on gene expression and post thaw sperm quality of Pacific abalone. Two ions, Na+ (459.1 ± 3.1 mM) and Cl– (515.9 ± 1.1 mM), were predominant in the seminal plasma (pH: 6.8 ± 0.1; osmolarity: 1,126 ± 3 mOsmL–1). Cryopreservation reduced mRNA expression levels of protein kinase A (PKA-C) and heat shock proteins (HSP70 and HSP90) genes in sperm. Fluorescent technique was used to compare morphological defects, acrosome integrity (AI), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), and DNA integrity of sperm cryopreserved with five different cryopreservation solutions (8% Me2SO, 8% EG, 6% PG, 2% GLY, and 2% MeOH). Droplet in tail and coiled tail defects was not observed for sperm cryopreserved with 8% Me2SO or 2% GLY. Sperm cryopreserved with 8% Me2SO showed improved DNA integrity and lower cryodamage than sperm cryopreserved with other cryoprotectants. Sperm to egg ratio of 10,000:1 was found to be the most suitable ratio for in vitro fertilization among different ratios tested. The fertilization rate of sperm cryopreserved with 8% Me2SO was not significantly (p &gt; 0.05) different from that of sperm cryopreserved with 2% GLY. DNA fragmentation showed strongly negative relationships with sperm quality parameters. Sperm cryopreserved with 8% Me2SO showed higher post thaw quality and mRNA expression of sperm motility associated gene than those cryopreserved with other cryoprotectants. The present research suggests to use 8% Me2SO for cryopreservation of Pacific abalone sperm as well as for hatchery production.

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