RESPONSE OF TURMERIC EXPLANT ON CYTOKININ AND AUXIN IN MURASHIGE AND SKOOG

2019 ◽  
Vol 24 (1) ◽  
pp. 1-12
Author(s):  
Mita Indriani ◽  
Erni Suminar ◽  
Noladhi Wicaksana ◽  
Denny Sobardini ◽  
Sulistyaningsih Sulistyaningsih ◽  
...  
Keyword(s):  
Tissue Culture ◽  
T Test ◽  
Test Method ◽  
Seed Technology ◽  
Planting Material ◽  
Randomized Design ◽  
Student’S T ◽  
And Control ◽  
Student’S T Test

This study was aimed at determining the concentration of several types of cytokinins and auxin for the induction of turmeric shoots in vitro. The research was conducted at the Tissue Culture Seed Technology Laboratory, Faculty of Agriculture, Padjadjaran University, Jatinangor. The study was conducted from October 2017 to February 2018. The source of planting material is in the form of shoots from the turmeric rhizome. The source of explants or planting material came from the field collected at the Tissue Culture Seed Technology Laboratory, Faculty of Agriculture, Padjadjaran University. Explants were taken from rhizome buds with a size of 0.6-2.0 cm. The experiment used a Completely Randomized Design which was analyzed using the Student’s T-test method. The number of experimental and control groups in this study were seven groups. Variation in treatment with different BAP, thidiazuron, zeatin, and NAA concentrations in each group. The results show that Thidiazuron 1 mgL-1 + NAA 1 mgL-1 gives better results on the percentage of live explants and number of shoots on turmeric plants (Curcuma domestica Val.) Clones 41 at the age of 14 weeks after planting.RESPONS EKSPLAN KUNYIT PADA SITOKININ DAN AUKSIN DALAM MEDIA MURASHIGE DAN SKOOGPenelitian ini bertujuan untuk mendapatkan salah satu konsentrasi dari beberapa jenis sitokinin dan auksin untuk induksi tunas kunyit secara in vitro. Penelitian dilakukan di Laboratorium Kultur Jaringan Teknologi Benih, Fakultas Pertanian, Universitas Padjadjaran, Jatinangor. Waktu pelaksanaan penelitian ini dimulai pada awal bulan Oktober 2017 sampai bulan Februari 2018. Sumber bahan tanam berupa tunas dari rimpang tanaman kunyit. Sumber eksplan atau bahan tanam berasal dari lapangan yang dikoleksi di Laboratorium Kultur Jaringan Teknologi Benih, Fakultas Pertanian, Universitas Padjadjaran. Eksplan diambil dari mata tunas rimpang dengan ukuran 0,6-2,0 cm. Percobaan menggunakan Rancangan Acak Lengkap yang dianalisis menggunakan metode Student’s T-test. Jumlah kelompok eksperimen dan kontrol dalam penelitian ini adalah tujuh kelompok. Variasi perlakuan dengan penambahan konsentrasi BAP, thidiazuron, zeatin, dan NAA yang berbeda pada setiap kelompok. Hasil penelitian ini menunjukkan bahwa Thidiazuron 1 mgL-1 + NAA 1 mgL-1 memberikan hasil yang lebih baik pada persentase eksplan hidup dan jumlah tunas pada tanaman kunyit (Curcuma domestica Val.) klon 41 pada umur 14 MST (Minggu Setelah Tanam).

18 EMBRYO AGGREGATION IN PIG IMPROVES CLONING EFFICIENCY AND EMBRYO QUALITY

2016 ◽  
Vol 28 (2) ◽  
pp. 139
Author(s):  
C. Buemo ◽  
A. Gambini ◽  
L. Moro ◽  
R. F. Y. Martin ◽  
D. Salamone
Keyword(s):  
In Vitro Culture ◽  
Dna Fragmentation ◽  
Embryo Development ◽  
Embryo Quality ◽  
T Test ◽  
Control Group ◽  
Reconstructed Embryo ◽  
Student’S T ◽  
Student’S T Test

In this study, we analysed the effects of the cloned embryo aggregation on in vitro embryo development and embryo quality by measuring blastocyst size and cell number, DNA fragmentation levels by TUNEL assay, and the relative expression of genes associated with pluripotency, apoptosis, trophoblast markers, and DNA methylation in the porcine. Cumulus-oocyte complexes were recovered from slaughterhouse ovaries by follicular aspiration. Maturation was performed in TCM for 42 to 48 h at 39°C and 5% CO2. After denudation by treatment with hyaluronidase, mature oocytes were stripped of the zona pellucida using a protease and then enucleated by micromanipulation; staining was performed with Hoëchst 33342 to observe metaphase II. Ooplasms were placed in phytohemagglutinin to permit different membranes to adhere between each other; the ooplasm membrane was adhered to a porcine fetal fibroblast from an in vitro culture. Adhered membranes of the donor cell nucleus and enucleated oocyte cytoplasm were electrofused through the use of an electric pulse (80 V for 30 μs). All reconstituted embryos were electrically activated using an electroporator in activation medium (0.3 M mannitol, 1.0 mM CaCl2, 0.1 mM MgCl2, and 0.01% polyvinyl alcohol) by a DC pulse of 1.2 kVcm for 80 μs. Then, embryos were incubated in 2 mM 6-DMAP for 3 h. In vitro culture of zona-free embryos was achieved in a well of wells system in 100 μL of SOF medium. Two experimental groups were used, one control group with a single reconstructed embryo per microwell (1×) and the other group placing 3 reconstructed embryo per microwell (3x aggregation group). Embryos were cultivated at 39°C in 5% O2, 5% CO2 for 7 days in SOF medium with a supplement of 10% fetal bovine serum on the fifth day. At Day 7, resulting blastocysts were classified according to their morphology and diameter to determine their quality. Our results showed that aggregation of 3× embryos increased blastocyst formation rate and blastocyst size of pig cloned embryos (Fisher’s test P < 0.05 and Student’s t-test P < 0.05, respectively). The DNA fragmentation levels in 3× aggregated cloned blastocysts were significantly decreased compared to 1x blastocyst (Student’s t-test P < 0.05). Levels of Oct4, Klf4, Igf2, Bax, and Dnmt1 transcripts were significantly higher in aggregated embryos, whereas Nanog levels were not affected. Transcripts of Cdx2 and Bcl-xl were essentially nondetectable (Student’s t-test P < 0.05). Our study suggests that embryo aggregation in the porcine may be beneficial for cloned embryo development and embryo quality, through a reduction in apoptotic levels and an improvement in cell reprogramming.

26 Drugs that Modify Epigenetics...What do they do to Porcine Clones?

2018 ◽  
Vol 30 (1) ◽  
pp. 152
Author(s):  
C. P. Buemo ◽  
A. Gambini ◽  
L. N. Moro ◽  
N. Canel ◽  
D. F. Salamone
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Electric Pulse ◽  
Donor Cell ◽  
T Test ◽  
Control Group ◽  
Developmental Rates ◽  
Student’S T ◽  
Student’S T Test

Although somatic cell nuclear transfer (SCNT) technology was developed more than 20 years ago, cloning efficiency remains low. Failures in the reprogramming of the donor cell result in embryos with aberrant epigenetic patterns and low developmental rates. In this study, we assessed whether the use the inhibitor of DNA (cytosine 5) methyltransferase 5-azacitidine (5Aza) combined with the MEK inhibitor in the MAPK pathway PD0325901 (PD) could improve SCNT efficiency in pigs. In vitro maturation of cumulus–oocyte complexes was performed in TCM for 44 h at 39°C under 5% CO2. Cumulus cells and zona pellucida was removed from matured oocytes, followed by enucleation of the metaphase plate previously stained with Hoëchst 33342. Each enucleated oocyte was attached to a donor cell by phytohemagglutinin treatment followed by an electric pulse of 80V for 30 μs. After fusion, reconstituted embryos were activated by an electric pulse followed by an incubation in 2 mM 6-DMAP for 3 h. Cloned embryos were cultured in vitro in a modified well-of-well system in SOF medium, where 3 cloned embryos were placed per microwell (3X). The experimental group 3X + drugs was exposed for the first 3 days to 1 μM PD and 1 μM 5Aza in SOF medium. After washing, embryos were cultured until Day 7 in regular SOF medium. The control group (3X) was cultured in regular SOF medium for 7 days. In vitro embryo developmental rates, gene expression, histone acetylation, and DNA methylation status were studied. The use of epigenetic modifying drugs significantly increased blastocyst rates (40.9% v. 29%; Fisher’s test, P < 0.05) and embryo size (41.46% v. 28.56%; Student’s t-test, P < 0.05) compared with the control group. Regarding gene expression, an increase of the relative expression of genes related to cell differentiation (Igf2 and Cdx2), antiapoptotic pathways (Bcl-xl) and DNA methylation modulation (Mapk1) was observed (P < 0.05). Pluripotency genes Oct4 and Nanog did not show differences between groups. The Bax proapoptotic gene significantly decreased its expression after drug treatment, as did the Klf4 gene (P < 0.05). Results were analysed by Student’s t-test. According to Histone H3K27ac, which is associated with enhancers or gene promoters, its marker was located mainly in the nuclear periphery respect to the control group with a uniform dispersion, indicating that the treatment could be activating certain genes by locating them near the periphery. Histone H3K4me1 was more uniformly localised throughout the nucleus in both groups. The intensity of the fluorescence was measured by quantitative confocal microscopy using a histogram produced by the ImageJ program (National Institutes of Health, Bethesda, MD, USA). Regarding DNA methylation by bisulphite sequencing, the 2 genes studied (Oct4 and DNMT1) showed a higher demethylation status for the treated group. Our results indicate that the combination of 5Aza+PD during early pre-implantation development dramatically increase blastocyst rates and embryo quality. This novel combination could be used as a strategy to improve the efficiency of SCNT in pigs and potentially other animals.

scholarly journals In vitro study of marginal, internal and proximal adaptation of implant-supported single-crown CAD/CAM restorations

2020 ◽  
Vol 19 ◽  
pp. e207286
Author(s):  
Kamila Aguiar Figueiredo Alves ◽  
Janaina Emanuela Damasceno ◽  
Viviane Maia Barreto de Oliveira ◽  
Luiz Gustavo Cavalcanti Bastos ◽  
Andrea Nóbrega Cavalcanti
Keyword(s):  
Repeated Measures ◽  
T Test ◽  
Digital Impression ◽  
Single Crown ◽  
Significant Difference ◽  
Cad Cam ◽  
Student’S T ◽  
One Way Anova ◽  
Student’S T Test

Aim: This study evaluated the precision of a CAD/CAM system by measuring marginal, internal and proximal fits in implantsupported single-crown restorations. Methods: Ten models of the upper arch were made in which implants replaced the upper left premolars. For fabrication of the zirconia infrastructures, titanium bases (TiBase) were coded and scanned using a scan body. A second digital impression was made for the fabrication of prostheses. Silicone impression material was used to determine the internal clearance between the TiBase and infrastructure and between the infrastructure and crown, whose thickness was measured at three points [P1 (cervical), P2 (middle) and P3 (occlusal)] with a stereoscopic microscope at 70x and 100x magnification. One-way ANOVA for repeated measures and the Student t-test were used for the analysis of internal and marginal adaptation. Proximal contacts were analyzed qualitatively. Results: There was no significant difference between the teeth evaluated (Student’s t-test; p>0.05) or between the corresponding points evaluated in either tooth (one-way ANOVA; p>0.05). Analysis of the internal clearance between the infrastructure and crown demonstrated that all points were significantly different compared to the reference standardized at 100 μm (Student’s t-test p<0.0001). There was no significant difference between P1 and P2, with the thickness at these two points being lower than that obtained at P3 (one-way ANOVA, p<0.05). The proximal contacts did not coincide with the quality defined by the device. Conclusion: The system tested was unable to produce implantsupported single-crown ceramic restorations with marginal, internal and proximal fits matching the digital workflow, with the inferior fits requiring adjustment prior to cementation.

scholarly journals Penambahan Glutathione pada Medium Fertilisasi Efektif Mendukung Pembentukan Pronukleus dan Perkembangan Awal Embrio Sapi (SUPPLEMENTATION OF GLUTATHIONE IN FERTILIZATION MEDIUM EFFECTIVELY SUPPORT NORMAL PRONUCLEUS FORMATION AND EARLY BOVINE EMBRYONIC DE

2017 ◽  
Vol 18 (3) ◽  
pp. 327
Author(s):  
Aras Prasetiyo Nugroho ◽  
Iman Supriatna ◽  
Mohamad Agus Setiadi
Keyword(s):  
Tissue Culture ◽  
Embryonic Development ◽  
Culture Medium ◽  
Fertilization Rate ◽  
Tissue Culture Medium ◽  
Randomized Design ◽  
Oviduct Fluid ◽  
And Control ◽  
Bovine Oocytes

The objective of this study was to determine fertilization rate effectiveness and early embryonic development competency with glutathione (GSH) supplementation in fertilization medium and culture This study consisted of two experiments comprising each of the four treatment and six repetitions with completely randomized design (CRD) using 651 oocytes. In the first experiment, a total of 317 bovine oocytes were matured in tissue culture medium (TCM) 199 at incubator 5% CO2 with temperature 39 ºC for24 h, then fertilized with sperm separated by swim up technique. Oocyte and sperms were incubated in fertilization medium supplemented with 0.25 mM, 0.50 mM, 1.00 mM GSH. In the second experiment, bovine oocytes were matured in maturation medium and fertilized with same procedure as mentioned before, then cultured in modified synthetic oviduct fluid (mSOF) with the following treatment: supplementation GSH only in fertilization medium (T1), supplementation GSH only in culture medium (T2), and supplementation GSH in both fertilization and culture medium (T3), while control not supplementation GSH (T0). Result of the first experiment showed that supplementation 1.00 mM GSH in fertilization medium can increase higher normal pronucleus (2PN) formation (86,9%) compared to other treatments, 0.50 mM (80.3%), 0.25 mM (73.8%), and control (58.9%) (P<0.05). In the second experiment showed that early bovine embryonic development on 2nd day cultured which reached 5-8 cell on treatment T1 (56.0%) and T3 (53.6%) were higher (P<0.05) compared to treatment T2 (26.2%) and T0 (control) (31.3%). Result of the other were showed that early bovine embryonic development on 4th day cultured which reached 9-16 cell on treatment T1 (26.2%) and T3 (27.4%) were higher (P<0.05) compared to that T2 (11.9%) and T0 (control) (10.8%). In conclusion, 1.00 mM GSH supplementation in medium was more effective in supporting normal pronucleus formation and early fertilization bovine embryonic development compared to in culture medium. ABSTRAK Penelitian ini bertujuan untuk mengetahui tingkat fertilisasi dan kompetensi perkembangan awal embrio sapi dengan penambahan glutathione (GSH) pada medium fertilisasi in vitro (IVF) dan kultur in vitro (IVC). Penelitian ini terdiri atas dua penelitian yang terdiri dari masing-masing empat perlakukan dan enam kali ulangan dengan rancangan acak lengkap (RAL) menggunakan 651 oosit. Penelitian I, sebanyak 317 oosit sapi dalam tissue culture medium (TCM) 199 dimatangkan pada inkubator 5% CO2 dan suhu 39°C selama 24 jam, kemudian difertilisasi dengan spermatozoa yang telah diseleksi menggunakan teknik swim up. Oosit dan spermatozoa diinkubasi pada medium fertilisasi dengan penambahan 0,25 mM, 0,50 mM, dan 1,00 mM GSH. Penelitian II, sebanyak 334 oosit sapi dimatangkan pada medium pematangan dan difertilisasi, kemudian dikultur pada medium modified synthetic oviduct fluid (mSOF), dengan perlakuan: penambahan GSH hanya pada medium fertilisasi (T1), penambahan GSH hanya pada medium kultur (T2), dan kombinasi penambahan GSH pada medium fertilisasi dan kultur (T3). Hasil penelitian I, menunjukkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi dapat meningkatkan pembentukan pronukleus normal (2PN) yang lebih tinggi (86,9%) dibandingkan dengan perlakuan yang lain yaitu 0,50 mM (80,3%), 0,25 mM (73,8%), dan kontrol (58,9%) (P<0,05). Penelitian II menujukkan bahwa perkembangan awal embrio sapi pada hari ke-2 kultur yang mencapai pembelahan 5-8 sel pada perlakukan T1 (56,0%) dan T3 (53,6%) lebih tinggi (P<0,05) dibandingkan dengan perlakuan T2 (26,2%) dan T0 (kontrol) (31,3%). Hasil penelitian lain menunjukkan bahwa perkembangan awal embrio sapi pada hari ke-4 kultur yang mencapai pembelahan 9-16 sel pada perlakuan T1 (26,2%) dan T3 (27,4%) lebih tinggi dibandingkan dengan perlakukan T2 (11,9%) dan T0 (kontrol) (10,8%) (P<0,05). Dapat disimpulkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi lebih efektif dalam mendukung pembentukan pronukleus normal dan perkembangan awal embrio sapi dibandingkan pada medium kultur.

scholarly journals Optimasi Konsentrasi Kinetin dan Benzyl Amino Purine Pada Kultur Tunas Vanili (Vanilla planifolia)

2021 ◽  
Vol 21 (1) ◽  
pp. 54-57
Author(s):  
Dyah Nuning Erawati ◽  
Yusriatul Mawaddah ◽  
Siti Humaida ◽  
Irma Wardati
Keyword(s):  
Tissue Culture ◽  
Shoot Multiplication ◽  
Culture Techniques ◽  
Planting Material ◽  
Randomized Design ◽  
Wet Weight ◽  
The Mean ◽  
Completely Randomized Design ◽  
Tissue Culture Techniques

Vanilla has a potential to be developed through tissue culture techniques to anticipate the limitations of the parent plant as a source of planting material. The in vitro propagation ability of vanilla shoots needs to be controlled with the regulation of Kinetin and Benzyl Amino Purines. The interests of this study are 1) analysis of the response of vanilla explants at several Kinetin concentrations; 2) analysis of the response of vanilla explants at several concentrations of BAP and 3) analysis of the interaction of Kinetin and BAP on the response of vanilla explants to form shoot multiplication. The research was conducted at the Tissue Culture Laboratory Politeknik Negeri Jember from June to December 2020 using a factorial Completely Randomized Design (CRD). Factor 1 was the Kinetin concentration of 0.0, 1.0, 2.0 mg.L-1 and the second factor was the concentration of BAP 0.5, 1.5, 2.5 mg.L-1. The results proved that the fastest shoot multiplication occurred on MS medium + Kinetin 2 mg.L-1 with a mean of 8.7 days after inoculation. The mean number of shoots was 7.6 shoots/explant with the highest average wet weight of 0.9 grams/explant at the addition of BAP 1.5 mg. L-1 at measurement 70 days after inoculation.

scholarly journals Pertumbuhan dan Perkembangan Jaringan Meristem Bawang Merah (Allium ascalonicum L.) Kultivar Katumi Secara In Vitro

Jurnal Agro ◽  
10.15575/1344 ◽  
2017 ◽  
Vol 4 (2) ◽  
pp. 97-109
Author(s):  
Lamro Purba ◽  
Erni Suminar ◽  
Denny Sobardini ◽  
Wieny Rizky ◽  
Syariful Mubarok
Keyword(s):  
Tissue Culture ◽  
Leaf Number ◽  
Shoot Induction ◽  
Seed Technology ◽  
Randomized Design ◽  
Rooting Media ◽  
Completely Randomized Design ◽  
Four Levels ◽  
Induction Stage

This study aimed for knowing and obtaining the best concentration of kinetin and NAA interaction effects in influencing the shoot induction, knowing how the plant growth regulators in induction mediastill affect the shoot additionin the MS0media and also knowing the largest number of roots in rooting media for shallot by in vitro. The experiment was conducted at Laboratory of Tissue Culture Seed Technology, Faculty of Agriculture, Padjadjaran University, during January 2011 until May 2011. This experiment divided in 3 stages, namely shoot induction stage, shoot subculture to MS0 media stage and shoot subculture to rooting media stage. Experimental method used in the shoot induction stage was factorial Completely Randomized Design with three replications. The first factor was the kinetin with four levels,0, 1, 2, and 3 mg L-1. The second factor was the NAA with three levels, as 0, 0.01, and 0.1 mg L-1. Basic media used for each treatment was MS. The experiment result showed there was an interaction between kinetin and NAA on shoot induction stagewith the plantlet height, leaf number, and shoot addition. The best result for leaf number was gained from interaction with 2 mg L-1 kinetin without NAA,while the treatment of 2 mg L-1 kinetin with 0.01 mg L-1 NAA gave a better interaction for theshoot addition variable.

scholarly journals Multiplikasi in vitro stroberi kultivar Tochiotome dengan penambahan jenis dan konsentrasi sitokinin untuk perbanyakan bibit

Kultivasi ◽  
2020 ◽  
Vol 19 (3) ◽  
Author(s):  
Ega Raisya ◽  
Denny Sobardini Sobarna ◽  
Anne Nuraini ◽  
Syariful Mubarok ◽  
Erni Suminar ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Fast Time ◽  
Endogenous Auxin ◽  
Seed Technology ◽  
Randomized Design ◽  
Process Plants ◽  
Number Of Leaves ◽  
Completely Randomized Design

Sari Perbanyakan tanaman stroberi secara konvensional dilakukan dengan menggunakan stolon, tetapi kurang efektif serta kualitas bibit yang dihasilkan kurang baik akibat adanya akumulasi penyakit. Budidaya stroberi memerlukan adanya perbanyakan bibit secara massal, tetapi tidak mengubah kualitasnya. Multiplikasi in vitro menjadi solusi untuk penyediaan bibit berkualitas dalam jumlah besar. Upaya untuk mendapatkan tunas in vitro dalam jumlah banyak yakni perlu adanya penambahan zat pengatur tumbuh golongan sitokinin seperti Benzylaminopurine (BAP) atau Thidiazuron (TDZ). Tujuan penelitian ini adalah mengetahui dan menetapkan jenis serta konsentrasi sitokinin dengan hasil terbaik dalam multiplikasi stroberi kultivar Tochiotome. Percobaan dilaksanakan di Laboratorium Kultur Jaringan Tanaman, Teknologi Benih, Fakultas Pertanian, Universitas Padjadjaran. Penelitian menggunakan Rancangan Acak Lengkap yang terdiri dari tujuh perlakuan yang diulang lima kali, yaitu: Kontrol (tanpa sitokinin); BAP (0,25 ppm; 0,50 ppm; 0,75 ppm), dan TDZ (0,25 ppm; 0,50 ppm; 0,75 ppm). Hasil dari percobaan menunjukkan bahwa penambahan sitokinin tidak berpengaruh nyata terhadap jumlah tunas dan bobot segar planlet. Media perlakuan kontrol dapat menghasilkan jumlah akar lebih banyak dibandingkan dengan media ditambah sitokinin. Penambahan BAP 0,50 ppm  berpengaruh positif terhadap jumlah daun dan dapat menghasilkan runner secara in vitro. Pemberian BAP 0,50 ppm cenderung dapat meningkatkan dan mempercepat produksi bibit tanaman stroberi kultivar Tochiotome.Kata Kunci: Benzylaminopurine (BAP), Thidiazuron (TDZ), Stroberi, Kultur Jaringan AbstractStolon is used for conventional propagation of strawberry, but it is less effective and the quality of the seeds is not good due to the accumulation of disease. In vitro multiplication becomes a solution for the supply of quality seeds in a fast time. The addition of growth regulator cytokinin, such as Benzylaminopurine (BAP) or Thidiazuron (TDZ) can produced the large number of shoot. The objective of this study was to obtain the best type and concentration of cytokinin in the multiplication of strawberry ‘Tochiotome’. The study was conducted at the Plant Tissue Culture Laboratory, Seed Technology, Faculty of Agriculture, Universitas Padjadjaran. This study used a Completely Randomized Design (CRD) with seven treatments and five replications, that were: Control (without cytokinin); BAP (0.25 ppm; 0.50 ppm; 0.75 ppm), and TDZ (0.25 ppm; 0.50 ppm; 0.75 ppm). The results indicated that addition of cytokinin did not affected increasing number of shoots and fresh weightof plantlets. Control media can produce larger number of roots than those containing PGRs, this might be due to the endogenous auxin concentrations found in strawberry plants. Also, cytokinin inhibited root formations process. Plants treated with BAP 0.50 ppm increased for the number of leaves and produced runners in vitro. This study showed application of BAP with 0.50 ppm increased and accelerated the production of strawberry ‘Tochiotome’ seedlings.Keywords: Benzylaminopurine (BAP), Thidiazuron (TDZ), Strawberry, Tissue Culture

scholarly journals In Vitro Screening Ketahanan Galur Padi (Oryza Sativa) B7 Hasil Rakitan Politeknik Negeri Lampung Terhadap Keracunan Besi (Fe)

2019 ◽  
Vol 19 (3) ◽  
pp. 241
Author(s):  
Onny Chrisna Pandu Pradana ◽  
Siti Novridha Andini
Keyword(s):  
Tissue Culture ◽  
Oryza Sativa ◽  
Iron Toxicity ◽  
In Vitro Screening ◽  
Culture Bottle ◽  
Randomized Design ◽  
Significant Difference ◽  
Completely Randomized Design ◽  
And Control

This research aimed to invstigate the response of paddy culture (B7 strain) assembled by Lampung State Polytechnic to the iron toxicity tolerance. The research was done at Plant Tissue Culture Laboratory, Lampung State Polytechnic, from July to September 2019. Treatments were single arranged in a completely randomized design with three replications. The treatment tried was five levels of Fe concentrations (5,6 ppm 28 ppm, 56 ppm, 84 ppm, 112 ppm, and control). Each replication consisted of three culture bottle containing one explant. The homogenity of data was tested using Barlett test. If the assumption were fulfilled then analysis of variance is executed using STATISTIX 10, and then followed by the Honest Significant Difference (HSD) test in 5% alpha for mean separation and RPA analysis. The result of this research showed that the B7 strain has tolerance to iron toxicity until 56 ppm of Fe concentration, it can be concluded from the PAR value of its strain (>0,50). Meanwhile in  84 and 112 ppm of Fe concentration, the RPA value of B7 starin (<0,50), and it is indicate that its strain is sensitive. 

RESPONSE OF TURMERIC SHOOT EXPLANT AFTER CYTOKININ WITH IN VITRO

2019 ◽  
Vol 24 (2) ◽  
pp. 143-152
Author(s):  
Marhan Nurullia ◽  
Erni Suminar Suminar ◽  
Anne Nurani
Keyword(s):  
Tissue Culture ◽  
Root Length ◽  
Air Flow ◽  
T Test ◽  
Shoot Height ◽  
Best Response ◽  
Randomized Design ◽  
Completely Randomized Design ◽  
Laminar Air Flow

This study was aimed at determining the response of turmeric shoot explants after the provision of various types and concentrations of cytokinins in vitro. This experiment was conducted at the Tissue Culture Laboratory, Faculty of Agriculture, Padjadjaran University from January to April 2018. The data were analyzed using T-Test. The experimental method used in this research was Completely Randomized Design (CRD). Explant planting was carried out in Laminar Air Flow. The experiment consisted of 7 treatments consisting of 4 replications and each test consisted of 4 units. Observation of this experiment was carried out for 12 MST. The main observations were made on the data that were tested statistically namely the percentage of explant growing shoots, percentage of explant growing roots, shoot height, number of tuns, number of roots and root length. The treatments consisted of Control, 2.5 mg L-1 BAP, 5 mg L-1 BAP, 0.5 mg L-1 TDZ, 1 mg L-1 TDZ, 0.01 mg L-1 Zeatin and 0.1 mg L Zeatin -1. The results show that the treatment of 1 mg L-1 TDZ shows the best response to the growth of turmeric explants by increasing the number of turmeric shoot explants than the others.RESPONS EKSPLAN TUNAS KUNYIT SETELAH SITOKININ SECARA IN VITROTujuan dari penelitian ini yaitu untuk melihat respons eksplan tunas kunyit terhadap pemberian berbagai jenis dan konsentrasi sitokinin secara in vitro. Percobaan ini dilakukan di Laboratorium Kultur Jaringan, Fakultas Pertanian, Universitas Padjadjaran dari bulan Januari sampai April 2018. Hasil percobaan dianalisis dengan Sample T-Test. Metode percobaan yang digunakan dalam penelitian ini yaitu Rancangan Acak Lengkap (RAL). Penanaman eksplan dilakukan di dalam Laminar Air Flow. Percobaan terdiri dari 7 perlakuan sebanyak 4 ulangan dan setiap ulangan terdiri dari 4 unit. Pengamatan percobaan ini dilakukan selama 12 MST. Pengamatan utama dilakukan terhadap data-data yang diuji secara statistik yakni persentase eksplan tumbuh tunas, persentse eksplan tumbuh akar, tinggi tunas, jumlah tunas, jumlah akar dan panjang akar. Perlakuan terdiri dari Kontrol; 2,5 mg L-1 BAP; 5 mg L-1 BAP; 0,5 mg L-1 TDZ; 1 mg L-1 TDZ; 0,01 mg L-1 Zeatin; dan 0,1 mg L-1 Zeatin. Hasil penelitian menunjukkan bahwa perlakuan 1 mg L-1 TDZ menunjukkan respons yang lebih baik terhadap pertumbuhan eksplan kunyit dengan meningkatkan jumlah tunas eksplan tanaman kunyit daripada yang lainnya.

Over Expression of MN1 Confers Resistance to Chemotherapy In Vitro and In Vivo

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 969-969
Author(s):  
Timothy Pardee ◽  
Teresa Mascenik ◽  
Britt H. Bolemon ◽  
Guerry J Cook
Keyword(s):  
Bone Marrow ◽  
The United States ◽  
Genetic Alterations ◽  
T Test ◽  
Over Expression ◽  
Student’S T ◽  
Worse Prognosis ◽  
Student’S T Test

Abstract Abstract 969 Acute myeloid leukemia (AML) is an accumulation of immature myeloid precursors that leads to progressive marrow failure and death. This disease affects approximately 12,000 people per year in the United States, causing 9,000 deaths. Despite decades of active research the overall 5 year survival remains a dismal 30–40%. The backbone of initial therapy for the last 30 years is combination chemotherapy containing cytarabine (Ara-C) and an anthracycline. Resistance to these therapies is a major problem and most patients diagnosed with AML will ultimately die from resistant disease. AML is characterized by heterogeneous genetic alterations that can be used to delineate prognosis. Using standard karyotyping techniques patients can be divided into good, intermediate and poor prognostic categories. There is a clear link between these chromosomal aberrations and response to chemotherapy as complete remission rates are significantly different between groups. Patients with no detectable cytogenetic abnormality fall into an intermediate prognostic group with a very heterogeneous outcome. Recent work has begun to uncover submicroscopic genetic alterations that effect prognosis for these patients. These alterations can be mutations, over or under expression of a particular gene. The MN1 gene encodes a transcription co-factor first identified by its involvement in a balanced translocation in a patient with a meningioma. Since its initial description it has been found over-expressed in multiple AML patient samples. There are several reports that over-expression of MN1 confers a worse prognosis in AML. High MN1 expressers were less likely to achieve a remission and had a lower 3 year survival rate. Additionally, over expression of MN1 in murine bone marrow leads to AML in transplanted recipients and predicts for resistance to ATRA in elderly AML patients. However, the effect of MN1 over expression on response to standard chemotherapy is currently unknown. To answer this question we used a murine model of AML driven by MLL-ENL. AML blasts were infected with retroviral vectors that contained MN1 and a GFP reporter. Partially infected blast populations were then exposed to various concentrations of either Ara-C or doxorubicin and the ratio of GFP positive and negative cells was compared to untreated controls. When blasts were exposed to 150 nM Ara-C the GFP+ percentage went from 21.10 (+/− 0.5302) in the control samples to 35.68 (+/−1.230) in the treated samples. This result was even more profound when cells were treated with 15 ng/ml doxorubicin where the percentage went from 21.10 (+/− 0.5302) to 80.27 (+/−1.615). Both results were highly statistically significant by two tailed student's t test with p values of 0.004 and <0.0001 respectively. Consistent results were obtained in multiple different infections and with separately derived MLL-ENL lines. These data demonstrate that blasts expressing MN1 had an advantage when exposed to either Ara-C or doxorubicin although the effect was far more pronounced with doxorubicin exposure. MN1 expressing blasts were also resistant to the combination of Ara-C and doxorubicin. In order to determine if MN1 conferred resistance to Ara-C and doxorubicin in vivo we injected sublethally irradiated, Ly5.1+ C57Bl6 recipients with a partially infected population of blasts. Ly5.1+ animals do not express the Ly5.2 allele; thus, staining cells for Ly5.2 allows differentiation of leukemic cells from endogenous marrow. Eight days after injection of blasts animals were treated with 100 mg/kg Ara-C plus 3 mg/kg doxorubicin daily for 5 days or observed. On day 6 animals were sacrificed and bone marrow from bilateral femurs was harvested, stained for Ly5.2 and analyzed by flow cytometery. Animals treated with Ara-C plus doxorubicin had 90.58% (+/−0.6638) Ly5.2+, GFP+ blasts compared to 55.38% (+/−5.245) in control animals. This result was highly statistically significant with a p value of <0.0001 by two tailed student's t test. This observation was reproducible in a separately derived MLL-ENL driven cell line. These data suggest that over expression of MN1 in this murine AML model confers resistance to both Ara-C and doxorubicin in vitro and in vivo and provides a biological explanation for the clinical observation that it confers a worse prognosis. The mechanisms involved in this resistance are currently under study. Disclosures: No relevant conflicts of interest to declare.

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