scholarly journals Transformasi Genetik Tanaman Kentang (Solanum tuberosum L.) dengan Gen acvB Menggunakan Vektor Agrobacterium tumefaciens

2021 ◽  
Vol 11 (1) ◽  
pp. 63
Author(s):  
DARWIN SILALAHI ◽  
I GEDE PUTU WIRAWAN ◽  
MADE SRITAMIN

Agrobacterium tumefaciens Mediated Genetic Transformation of acvB Gene in Potato (Solanum tuberosum L.). Genetic transformations are now routinely applied to plant mediated by Agrobacterium tumefaciens as the most convenient technique. This study aimed to prove the success of A. tumefaciens mediated genetic transformation in potato. A. tumefaciens LBA (pBI 121) and explant of potato shoot were used in this study. Explants were grown in vitro on Murashige and Skoog media. Transformation was implemented using smear technique by smearing A. tumefaciens to injured explant. Experimental groups consisted of two groups: control group which did not receive transformation treatment and treatment group receiving transformation treatment. Explant growth was observed through the presence of shoots, branches and the shoot height. Explants in the treatment group resulted in a higher number of shoots, branches, and shoot heights compared to control. Phenol compounds appear in explant epidermal tissue, indicating the wounds produced by A. tumefaciens infection, thus the gene predicted to be transformed. Identification by PCR is needed to prove the existence of the acvB gene in potato plants genome, using acvB specific PCR primer as the marker, such as (5?-CCCT CTAG AGAC CCGC GCCA AGGCG-3?) and (5?CGCG TCGA CCTT GTCG GAAAG -3?) with 540-bp in base pair size produced.

1970 ◽  
Vol 20 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Rita Sarah Borna ◽  
M. I. Hoque ◽  
R. H. Sarker

Genetic transformation using nodal and internodal segments from three economically important potato (Solanum tuberosum L.) varieties namely, Diamant, Cardinal and Granola was conducted using an Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI12 containing the GUS and nptII genes. Node and internodal segments were used for direct regeneration as well as regeneration with the intervention of callus. best responses were  obtained for direct regeneration of shoots when the explants were cultured on MS supplemented with 4.0 mg/l BAP +1.0 mg/l IAA, 1.5 mg/l BAP  + 0.5 mg/l IAA and 5.0 mg/l BAP +1.0 mg/l IAA in Diamant, Cardinal  and  Granola, respectively. In Diamant spontaneous in vitro microtuberization was obtained from these proliferated shoots. Further culturing of these in vitro grown green microtubers regenerated a large number of shoots on MS containing 4.0 mg/l BAP +1.0 mg/l IAA. By combining the best treatments, this protocol yielded an average transformation rate of 87% of treared explants. Stable expression of GUS gene was visualized in the various parts of transformed shoots through histochemical assay. Genomic DNA was isolated from transformed shoots and stable integration of the GUS and nptII genes was confirmed by PCR analysis.   Key words:  Potato, in vitro regeneration, transformation   D.O.I. 10.3329/ptcb.v20i2.6894   Plant Tissue Cult. & Biotech. 20(2): 145-155, 2010 (December)


1996 ◽  
Vol 12 (2) ◽  
pp. 42-46 ◽  
Author(s):  
P. G. Kovalenko ◽  
I. M. Yefimenko ◽  
N. V. Schuman ◽  
T. V. Medvedeva ◽  
K. G. Gazaryan ◽  
...  

2000 ◽  
Vol 18 (1) ◽  
pp. 41-45 ◽  
Author(s):  
Antonio Carlos Torres ◽  
Adriana Teixeira Ferreira ◽  
Eduardo Romano ◽  
Mônica Kangussú Cattony ◽  
Adriana Souza Nascimento

Explantes de segmentos nodais de batata (Solanum tuberosum L.) cultivar Achat provenientes da cultura in vitro foram transformados, via Agrobacterium tumefaciens, estirpe LBA4404, contendo o vetor binário pBI121 com os genes npt II e o gus-intron. A regeneração de potenciais transformantes foi feita em meio seletivo contendo 50 mg.L-1 de canamicina. Foram obtidas plantas expressando resistência à canamicina e atividade da b-glucuronidase. Pela análise molecular nas plantas gus+, via PCR, constatou-se a amplificação dos genes npt II. A hibridização por Southern blotting demonstrou a inserção de pelo menos uma cópia do gene introduzido.


2017 ◽  
Author(s):  
Yunita Prameswari ◽  
FNU Djenal ◽  
FNU Damanhuri

Kebutuhan kentang yang semakin tinggi menyebabkan permintaan semakin meningkat. Rendahnya produksi kentang mengakibatkan berbagai upaya untuk peningkatan produksi terus dilakukan. Penggunaan metode kultur jaringan yaitu metode untuk mengisolasi bagian tanaman seperti protoplasma, sel, sekelompok sel, jaringan dan organ dalam kondisi aseptik, sehingga dapat diperbanyak dan beregenerasi menjadi tanaman utuh dapat dijadikan alternatif pemenuhan kebutuhan. Penelitian ini bertujuan untuk mengetahui kecepatan pembentukan umbi mikro kentang (Solanum Tuberosum L.) varietas granola kembang secarain vitro dengan menggunakan dua faktor dan 3 kali ulangan. Faktor pertama yaitu aspirin dengan tiga taraf (5,10,15) ppm. Faktor kedua yaitu kinetin dalam tiga taraf (8,10,12) ppm. Penelitian menggunakan seluruh propagul kentang yang berumur 30 hari setelah subkultur dan data yang didapat dianalisis menggunakan ANOVA. Hasil penelitian menunjukan bahwa interaksi aspirin dan kinetin tidak berpengaruh terhadap jumlah akar, kedinian umbi, dan bobot umbi. Interaksi perlakuan terbaik bagi pembentukan tunas yaitu A2K1 aspirin 10 ppm dan kinetin 8 ppm sedangkan Interaksi perlakuan terbaik pada parameter jumlah umbi yaituA3K2 aspirin 15 ppm dan kinetin 10.


2019 ◽  
Vol 9 (2) ◽  
pp. 91
Author(s):  
Ghea Dotulong ◽  
Stella Umboh ◽  
Johanis Pelealu

Uji Toksisitas Beberapa Fungisida Nabati terhadap Penyakit Layu Fusarium (Fusarium oxysporum) pada Tanaman Kentang (Solanum tuberosum L.) secara In Vitro (Toxicity Test of several Biofungicides in controlling Fusarium wilt (Fusarium oxysporum) in Potato Plants (Solanum tuberosum L.) by In Vitro) Ghea Dotulong1*), Stella Umboh1), Johanis Pelealu1), 1) Program Studi Biologi, FMIPA Universitas Sam Ratulangi, Manado 95115*Email korespondensi: [email protected] Diterima 9 Juli 2019, diterima untuk dipublikasi 10 Agustus 2019 Abstrak Tanaman kentang (Solanum tuberosum L.) adalah salah satu tanaman hortikultura yang sering mengalami penurunan dari segi produksi dan produktivitasnya, akibat adanya serangan penyakit layu yang salah satunya disebabkan oleh Fusarium oxysporum. Tujuan penelitian ini adalah mengidentifikasi toksisitas beberapa fungisida nabati dalam mengendalikan penyakit Layu Fusarium (F. oxysporum) pada tanaman kentang (Solanum tuberosum L.) secara In Vitro. Metode Penelitian yang digunakan yaitu metode umpan beracun. Data dianalisis dengan Rancangan Acak Lengkap (RAL) dengan Analisis Varian (ANAVA) yang dilanjutkan dengan menggunakan metode BNT (Beda Nyata Terkecil). Hasil Penelitian, diperoleh nilai toksisitas fungisida nabati tertinggi yaitu pada ekstrak daun sirsak dengan nilai HR (69,44%), kategori berpengaruh, ditandai dengan diameter koloni 2,75 cm (100ppm) dan yang terendah toksisitasnya yaitu pada ekstrak daun jeruk purut dengan nilai HR (49,81%), kategori cukup berpengaruh ditandai dengan diameter koloni 3,75 cm (25ppm). Semakin tinggi konsentrasi yang diujikan maka semakin tinggi toksisitas dari fungisida nabati yang diberikan.Kata Kunci: fungisida nabati, Fusarium oxysporum, tanaman kentang, In Vitro Abstract Potato plants (Solanum tuberosum L.) is one of the horticulture plants which often decreases in terms of production and productivity, due to the attack of wilt, one of which is caused by Fusarium oxysporum. The purpose of this study was to determine the toxicity of several biofungicides in controlling Fusarium wilt (F. oxysporum) in potato plants (Solanum tuberosum L.) in Vitro. The research method used was the In Vitro method with the poison bait method. Data were analyzed by Completely Randomized Design with Variant Analysis (ANAVA), followed by the BNT method. The results showed that the highest biofungicide toxicity value was soursop leaf extract with HR values (69.44%), influential categories, characterized by colony diameter 2.75 cm (100ppm) and the lowest toxicity, namely in kaffir lime leaf extract with a value of HR (49.81%), quite influential category was characterized by colony diameter of 3.75 cm (25ppm). The higher the concentration tested, the higher the toxicity of the biofungicide given.Keywords: biofungicides, Fusarium oxysporum, Potato Plants, In Vitro.


Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Shiori Ashibe ◽  
Kanade Irisawa ◽  
Ken Yokawa ◽  
Yoshikazu Nagao

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.


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