• Title/Summary/Keyword: In-vitro techniques

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In Vitro Propagation of Medicinal Herbs in Korea

  • An, Chanhoon;Song, Jeongho
    • Journal of Forest and Environmental Science
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    • v.34 no.1
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    • pp.77-81
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    • 2018
  • Mass production of forest medicinal plants is related to quality control of raw medicinal materials. Plant tissue culture is an important technology to produce high-quality plant materials. Numerous factors are reported to influence the success of in vitro regeneration of medicinal plants. Embryogenesis is known to be the most effective techniques and it has developed in some medicinal plant species. Various in vitro cultural condition for direct and/or indirect somatic embryogenesis systems have developed in Epimedium koreaum, Bupleurum falcatum, Paeonia lactiflora, Chrysanthemum zawadskii, Houttuynia cordata etc. In this study, we provide the present statue and information of in vitro propagation techniques that is able to apply as an efficient system for rootstock propagation system of forest medicinal plants.

In Vitro Fertilization and Development of Bovine Oocytes (우 난포란의 체외수정과 발육)

  • 김정익
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.98-104
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    • 1989
  • Successful techniques of in vitro fertilization(IVF) are valuable for studying the process of fertilization and for developing economical procedures for gene and nuclear transfer in farm animals. To date, bovine IVF system has been developed with oocytes in vitro or vitro, but the resulting zygotes exhibit limited embryonic development after in vitro culture. Even though in vitro matured oocytes achieved high fertilization and cleavage rates, these embryos appear extremly low rate of pregnancies when transferred to synchronized recipients. Development of early bovine embryos in vitro is generally arrested at the 8-to 16-cell stage. However, recent use of somatic cells such as trophoblastic vesicle, granulosa and oviduct epithelial cell for co-culture with early bovine embryos has proven effective for development of embryos, matured and fertilized in vitro, past the in vitro cell blocks. These factors clearly indicate the value of the co-culture system in promoting development of bovine oocytes matured and fertilized in vitro to morula or blastocyst stage in vitro. In addition, co-culture system may beome a tool for evaluation of viability of ova that have been manipulated by procedures such as splitting, microinjection and nuclear transfer.

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Development Ability of Bovine Early Embryo Blastomere In Vitro in Embedding Matrix (체외생산 소 초기배 할구세포의 Embedding Matrix에서의 발생능력)

  • 이홍준;서승운;이상호;송해범
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.171-175
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    • 1995
  • This study was performed to establish the condition and the methods for the techniques of insertion the isolated blastomere cells into cytoplasm, in order to research the develop-mental ability of bovine embryo blastomere cells in vitro produced. After 24h in vitro ovary maturation with the ovaries from a slaughter house, in vitro fertilization was performed to the vital sperms which their mobility were decided by percoll gradient method, with 2~8 cell stage embryos, the blastomeres were isolated in $Ca^2$+. $Mg^2$+-free PBS, and following that embedded into agar and alginate solution, respectively. The rates of in vitro develop-ment are as follows ; in agar embedded 11 among 120(9.2%) 1 /2~1 /3 blastomers cleaved and 6 among 93(6.5%) 1 /4~1 /8 blastomeres cleaved. In sodium alginate-embedded 14 among 84(16.7%) 1 /2~1 /3 blastomeres cleaved and 6 among 85(7.1%) 1 /4~1 /8 blastomeres cleaved. In case of Na-alginate, the rate of the cells were better than those of agar. The results suggest that the techniques for embeeding the isolated blastomeres into gel may help cloning of bovine early embryo without nuclear transplantation.

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In vitro Plant Propagation: A Review

  • Kumar, Nitish;Reddy, M.P.
    • Journal of Forest and Environmental Science
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    • v.27 no.2
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    • pp.61-72
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    • 2011
  • Micropropagation is an alternative mean of propagation that can be employed in mass multiplication of plants in relatively shorter time. Recent modern techniques of propagation have been developed which could facilitate large scale production of true-to-type plants and for the improvement of the species using genetic engineering techniques in the next century. An overview on the in vitro propagation via meristem culture, regeneration via organogenesis and somatic embryogenesis is presented. The usefulness of the plants in commercial industry as well as propagation techniques, screening for various useful characteristics and the influence of different cultural conditions in the multiplication, rooting and acclimatization phases on the growth of tissue cultured plant discussed.

Predicting In Sacco Rumen Degradation Kinetics of Raw and Dry Roasted Faba Beans (Vicia faba) and Lupin Seeds (Lupinus albus) by Laboratory Techniques

  • Yu, P.;Egan, A.R.;Leury, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1377-1387
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    • 2000
  • Two laboratory techniques: (1) an in vitro method with two procedures for measuring protein degradabilities and (2) an in vitro method with three procedures for measuring protein solubility, were investigated to determine which laboratory techniques could most accurately predict the quantity of rumen protein degradation kinetics of legume seeds after dry roasting under various conditions, in terms of (1) rumen protein disappearance ($D_j$, where j=0, 2, 4, 8, 12, 24 and 48 h incubation), (2) rumen protein effective degradability (EDCP), (3) the parameters describing rumen degradation characteristics (the soluble fraction: S, the potentially degradable fraction: D, undegradable fraction: U, lag time: T0 and the degradation rate: Kd) and (4) rumen bypass protein (BCP), which were determined by the method accepted internationally at present, in sacco nylon bag technique using the standardized Dutch method. Feeds evaluated were the raw and dry roasted whole faba (Vicia faba) beans (WFB) and whole lupin (Lupinus albus) seeds (WLS), each was dry roasted under various conditions (at 110, 130 or $150^{\circ}C$ for 15, 30 or 45 min). In vitro protein degradability ($D_1$_Auf and $D_{24}$_Auf) were determined using the modified Aufr re method by enzymatic hydrolysis for 1 h and 24 h using a protease extracted from Streptomyces griseus in a borate-phosphate buffer. In vitro protein solubility ($bf_1$_S, $bf_2$_S, $bf_3$_S) was measured in a borate-phosphate buffer with three different procedures. Results from laboratory techniques (in vitro) were correlated and linearly regressed with in sacco results. Of the three procedures of in vitro protein solubility evaluated, none of them could predict in sacco results with good precision. The highest Pearson correlation coefficient ($R^2$) was less than 0.50. Of two procedures of in vitro protein degradability studied, the $D_1$_Auf values were closely correlated with in sacco parameters: Kd, EDCP and %BCP with high R' values: 0.82, 0.85 and 0.85, respectively, and closely correlated with in sacco $D_j$ at 2, 4, 8 and 12 h rumen incubation with high $R^2$ values: 0.83, 0.91, 0.93 and 0.83, respectively. The $D_{24}$_Auf values could not predict in sacco results. The highest $R^2$ value was less then 0.40. These results indicated that in vitro protein solubility measured in borate-phosphate failed to identify differences in the rate and extent of protein degradation of legume seeds after dry roasting under various conditions and thus should not be used to predict rumen degradation, particularly for heat processed feedstuffs. But in vitro protein degradability using the modified Aufr re method by enzymatic hydrolysis for 1 h or possibly an intermediate time (>1 h and <24 h) is a promising laboratory procedure to detect effectiveness of dry roasting legume seeds on rumen protein degradation characteristics and could be used as a simple laboratory method to predict the rate and extent of protein degradation in the rumen in sacco with high accuracy. The equations to predict EDCP, Kd and BCP of dry roasted legume seeds (WLS and WFB) under various conditions are as follow: For both: EDCP (%)=-1.37+1.06*$D_1$_Auf ($R^2=0.85$, p<0.01). For both: Kd (%/h)=-21.81+0.49*$D_1$_Auf ($R^2=0.82$, p<0.01). For both: %BCP=103.37-1.07*$D_1$_Auf ($R^2=0.85$, p<0.01).

Attempts to Transform Pollen Grains and Pollen Tubes in the Process of Fertilization in Tobacco

  • Chung Chan-Sun
    • Plant Resources
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    • v.8 no.2
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    • pp.87-90
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    • 2005
  • In tobacco, in vitro pollination has been successfully applied to overcome interspecific incompatibility. The use of this technique will make it possible to introduce DNA into pollen tubes just before fertilization. In this study, we showed improvement of the efficiency of in vitro self-pollination and introduction of foreign genes into pollen tubes by the method of polycation. A plasmid harbouring the GUS gene was introduced into pollen grains and pollen tubes, which had incubated on pollen germination medium(PGM), by polyornithine method. Transient expression of the GUS in pollen grains and pollen tubes that were treated with 0, 2, 5 and $10{\mu}g/m\ell$ DNA was observed. In results, combination of the techniques of polyornithine and in vitro pollination was efficient new technique for genetic transformation through fertilization processes.

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Current approaches for assisted oocyte maturation in camels

  • Saadeldin, Islam M.;Cho, Jongki
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.3
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    • pp.162-167
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    • 2021
  • Camel (camelus dromedarius) is a unique large mammalian species that can survive harsh environmental conditions and produce milk, meat, and wool. Camel reproduction is inferior when compared to other farm animal species such as cattle and sheep. Several trials have been reported to increase camel reproduction and production through assisted reproductive techniques (ARTs) such as in vitro fertilization and cloning. For these reasons, obtaining enough mature oocytes is a cornerstone for ARTs. This demand would be improved by the oocyte in vitro maturation (IVM) systems. In this review, the current approaches and views from different laboratories using ARTs and the IVM to produce embryos in vitro in camel species. For the last two decades, conventional IVM system was the common approach, however, recently the bi-phasic IVM system has been introduced and showed promising improvement in IVM of camel oocytes. Detailed studies are needed to understand camel meiosis and IVM to efficiently increase the production of this species.

Foliar Micromorphological Response of In Vitro Regenerated and Field Transferred Plants of Oldenlandia umbellata L.: A Medicinal Forest Plant

  • Jayabal, Revathi;Rasangam, Latha;Mani, Manokari;Shekhawat, Mahipal Singh
    • Journal of Forest and Environmental Science
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    • v.35 no.1
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    • pp.54-60
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    • 2019
  • Plant tissue culture techniques offer quick methods of regeneration of plants of medicinal importance but the survival chances of such plants are always questionable when shifted to the in vivo conditions. The present study enumerates the micromorphological developments in the leaves of in vitro regenerated and field transferred plantlets of Oldenlandia umbellata. The leaves developed in vitro after $4^{th}$ subcultures of multiplication phase and after 6 weeks of field transferred plants were used. Statistically significant differences in the number of stomata, veins, raphides, crystals and trichome density per square mm were observed. The improvements in stomatal apparatus and density (decreased from 41.85 to 32.20), developments in leaf architectural parameters and emergence of defense mechanism through increased numbers of raphides (8 to 15), crystals and trichomes (13.5 to 18.2) proved acclimation of tissue culture raised plantlets from in vitro to the in vivo environments lead to 100 % success in field establishment of the plantlets. The in vitro induced foliar abnormalities (changes in stomata, venation pattern, vein density, trichomes, crystals etc.) were repaired while hardening of plantlets in the greenhouse and finally in the field. The observed micromorphological response of leaves under altered environmental conditions could help in determination of proper stage of field transfer and prediction of survival percentage of in vitro regenerated O. umbellata plantlets.

Evaluation and Method of In Vitro Digestibility in Monogastric Animal Model (단위동물 모델에서 In vitro 소화율 측정과 평가)

  • Kang, Lyeongin;Kim, Jin Seon;Lee, Sung Sill;Chu, Gyo-Moon;Kim, Jin Wook
    • Journal of agriculture & life science
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    • v.53 no.2
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    • pp.15-26
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    • 2019
  • An in vitro digestibility methods have been developed to mimic an in vivo system in the past decades. Because the in vivo techniques cause high cost, intensive labor, longer research periods and ethnical problems. In this review, the digestive systems from stomach to large intestine of pig as a monogastric animal were addressed to understand an in vivo digestion. The innovative in vitro technique using the Daisy II incubator was performed and the in vitro ileal and fecal dry matter (DM) digestibility of corn, rice, wheat and barley was determined.