Slash pine genetic transformation through embryo cocultivation with A. tumefaciens and transgenic plant regeneration

2013 ◽  
Vol 50 (2) ◽  
pp. 199-209 ◽  
Author(s):  
Wei Tang ◽  
Bo Xiao ◽  
Yongjun Fei
Author(s):  
Masoumeh Nomani ◽  
Masoud Tohidfar

Abstract Background Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi. Results According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants. Conclusions In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25–35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.


Author(s):  
Richa Bajaj ◽  
Lani M. Irvin ◽  
Brajesh Nanda Vaidya ◽  
Sadanand A. Dhekney ◽  
Nirmal Joshee

2014 ◽  
Vol 50 (6) ◽  
pp. 696-706 ◽  
Author(s):  
Iqrar Ahmad Rana ◽  
Siegfried Salomon ◽  
Wilhelm Schäfer ◽  
Dirk Becker

2019 ◽  
Vol 2 (2) ◽  
pp. 77
Author(s):  
Zaiyin Rizky Ageng Maulidia ◽  
Wahyu Indra Duwi Fanata

ABSTRACT Indonesia is rich in rice genetic diversity in the form of javanica rice. Subspecies javanica have some superiority characters, among others a strong plant tissue, but in generally they have low productivity. Weaknesses of javanica rice can be repaired by biotechnology through genetic transformation techniques from callus of plant. Auxin types and genotypes are important factors for callus induction and plant regeneration. Therefore, this research held for know callus introduction and plant regeneration of three varieties of Javanica rice (Pendok, Genjah Arum, and Menthik Wangi Susu) combined with type of auxin (2,4-D and pychloram). The experimental design of RALs two factorials, there are three varieties of rice (Pendok, Genjah Arum, and Menthik Wangi Susu) and two types of auxin (2,4-D and Pychloram) and analyzed by DMRT α5%. The results of this study is Mentik Wangi Susu have highest regeneration than Pendok and Genjah Arum. The use of 2,4-D resulted in a higher callus induction and plant regeneration than pychloram in the three rice varieties used. While the best combination of treatments for callus induction and regeneration is Menthik Wangi Susu and 2,4-D. Keyword: auxin, javanica, callus induction, plant regeneration. ABSTRAK Indonesia kaya akan keragaman genetik padi berupa padi javanica. Padi sub spesies javanica memiliki beberapa keunggulan diantaranya jaringan tanaman yang kuat, namun pada umumnya mempunyai tingkat produktivitas gabah yang rendah. Kelemahan pada padi javanica dapat diperbaiki dengan bioteknologi melalui teknik transformasi genetik menggunakan jaringan kalus. Genotip dan jenis auksin merupakan faktor penting dalam induksi kalus dan regenerasi tanaman padi secara in vitro. Oleh karena itu, penelitian ini dilaksanakan untuk mengetahui daya pembentukan kalus dan regenerasi pada tiga varietas padi javanica, yaitu Pendok, Genjah Arum, dan Menthik Wangi Susu yang dikombinasikan dengan perlakuan jenis auksin berupa penggunaan 2,4-D dan pikloram. Rancangan percobaan yang digunakan adalah RAL dua faktorial yaitu varietas tanaman padi dan jenis auksin  dengan pengujian  menggunakan DMRT α5%. Hasil penelitian menunjukkan bahwa dari ketiga varietas padi yang digunakan, varietas Mentik Wangi Susu menunjukan daya regenerasi yang tertinggi dibandingkan varietas Pendok dan Genjah Arum.  Penggunaan 2,4-D menghasilkan tingkat pengkalusan dan regenerasi yang tinggi dibandingkan pikloram pada tiga varietas padi yang digunakan. Sedangkan kombinasi perlakuan yang terbaik untuk induksi kalus dan daya regenerasi adalah penggunaan varietas Menthik Wangi Susu dan auksin dalam bentuk 2,4-D.  Kata Kunci: auksin, javanica, induksi kalus, regenerasi tanaman. 


1996 ◽  
Vol 16 (1-2) ◽  
pp. 32-37 ◽  
Author(s):  
Sanchayita Kar ◽  
Tony M. Johnson ◽  
Pritilata Nayak ◽  
S. K. Sen

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