Construction of genetic transformation system of Salix mongolica: in vitro leaf-based callus induction, adventitious buds differentiation, and plant regeneration

2017 ◽  
Vol 132 (1) ◽  
pp. 213-217 ◽  
Author(s):  
Qingjie Guan ◽  
Mingling He ◽  
Haiyan Ma ◽  
Xu Liao ◽  
Zhenjuan Wang ◽  
...  
Keyword(s):  
Plant Regeneration ◽  
Genetic Transformation ◽  
Callus Induction ◽  
Adventitious Buds ◽  
Transformation System ◽  
Genetic Transformation System

scholarly journals PENGARUH JENIS AUKSIN TERHADAP PEMBENTUKAN KALUS DAN DAYA REGENERASI TIGA VARIETAS PADI LOKAL

2019 ◽  
Vol 2 (2) ◽  
pp. 77
Author(s):  
Zaiyin Rizky Ageng Maulidia ◽  
Wahyu Indra Duwi Fanata
Keyword(s):  
Genetic Diversity ◽  
Experimental Design ◽  
Plant Regeneration ◽  
Genetic Transformation ◽  
Callus Induction ◽  
Plant Tissue ◽  
Transformation Techniques ◽  
Rice Varieties ◽  
Rice Genetic

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. 

scholarly journals Standardized Genetic Transformation Protocol for Chrysanthemum cv. ‘Jinba’ with TERMINAL FLOWER 1 Homolog CmTFL1a

Genes ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 860
Author(s):  
Saba Haider ◽  
Yaohui Gao ◽  
Yike Gao
Keyword(s):  
Transgenic Plants ◽  
Genetic Transformation ◽  
Model Organism ◽  
Plant Morphology ◽  
Pcr Analysis ◽  
Transformation System ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Chrysanthemum X Morifolium

Chrysanthemum (Chrysanthemum x morifolium Ramat.) cultivar Jinba is a distinctive short-day chrysanthemum that can be exploited as a model organism for studying the molecular mechanism of flowering. The commercial value of Jinba can be increased in global flower markets by developing its proper regeneration and genetic transformation system. By addressing typical problems associated with Agrobacterium-mediated transformation in chrysanthemum, that is, low transformation efficiency and high cultivar specificity, we designed an efficient, stable transformation system. Here, we identify the features that significantly affect the genetic transformation of Jinba and standardize its transformation protocol by using CmTFL1a as a transgene. The appropriate concentrations of various antibiotics (kanamycin, meropenem and carbenicillin) and growth regulators (6-BA, 2,4-D and NAA) for the genetic transformation were determined to check their effects on in vitro plant regeneration from leaf segments of Jinba; thus, the transformation protocol was standardized through Agrobacterium tumefaciens (EHA105). In addition, the presence of the transgene and its stable expression in CmTFL1a transgenic plants were confirmed by polymerase chain reaction (PCR) analysis. The CmTFL1a transgene constitutively expressed in the transgenic plants was highly expressed in shoot apices as compared to stem and leaves. Overexpression of CmTFL1a led to a delay in transition to the reproductive phase and significantly affected plant morphology. This study will help to understand the biological phenomenon of TFL1 homolog in chrysanthemum. Moreover, our findings can explore innovative possibilities for genetic engineering and breeding of other chrysanthemum cultivars.

scholarly journals Establishment of a Plant Regeneration and Genetic Transformation System of Panax ginseng C.A. Meyer

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 572d-572
Author(s):  
Hak-Tae Lim ◽  
Haeng-Soon Lee ◽  
Tage Eriksson
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Genetic Transformation ◽  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Regeneration Systems

Plant regeneration of ginseng has been known to be difficult, and there are a few reports on plant regeneration of ginseng via somatic embryogenesis. In vitro flowering has, however, been one of the major drawbacks in these regeneration systems in which BA and GA3 were included in germination and shoot multiplication media. Multiplication of adventitious shoots from a single somatic embryo, abnormal morphology, and vitrified shoots were also observed. All these facts have made successful acclimatization of ginseng plantlets difficult. The purposes of this study were 1) to establish the plant regeneration system via organogenesis, 2) to improve normal plant regeneration via somatic embryogenesis, 3) to improve the efficiency of plant regeneration from protoplast culture, 4) to understand the acclimatization process, 5) to develop effective genetic transformation protocol. Data in relation with all these studies are presented in detail.

scholarly journals Establishment of a Genetic Transformation System in Guanophilic Fungus Amphichorda guana

2021 ◽  
Vol 7 (2) ◽  
pp. 138
Author(s):  
Min Liang ◽  
Wei Li ◽  
Landa Qi ◽  
Guocan Chen ◽  
Lei Cai ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Genetic Manipulation ◽  
Major Facilitator Superfamily ◽  
Protoplast Regeneration ◽  
Transformation System ◽  
Regeneration Rate ◽  
Bioactive Natural Products ◽  
Genetics Research ◽  
Genetic Transformation System ◽  
Major Facilitator

Fungi from unique environments exhibit special physiological characters and plenty of bioactive natural products. However, the recalcitrant genetics or poor transformation efficiencies prevent scientists from systematically studying molecular biological mechanisms and exploiting their metabolites. In this study, we targeted a guanophilic fungus Amphichorda guana LC5815 and developed a genetic transformation system. We firstly established an efficient protoplast preparing method by conditional optimization of sporulation and protoplast regeneration. The regeneration rate of the protoplast is up to about 34.6% with 0.8 M sucrose as the osmotic pressure stabilizer. To develop the genetic transformation, we used the polyethylene glycol-mediated protoplast transformation, and the testing gene AG04914 encoding a major facilitator superfamily transporter was deleted in strain LC5815, which proves the feasibility of this genetic manipulation system. Furthermore, a uridine/uracil auxotrophic strain was created by using a positive screening protocol with 5-fluoroorotic acid as a selective reagent. Finally, the genetic transformation system was successfully established in the guanophilic fungus strain LC5815, which lays the foundation for the molecular genetics research and will facilitate the exploitation of bioactive secondary metabolites in fungi.

scholarly journals Seed Germination and in vitro Plant Regeneration through Callus Culture of Two Lychnis Species

2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kee Hwa Bae ◽  
Eui Soo Yoon
Keyword(s):  
Plant Regeneration ◽  
Seed Germination ◽  
Callus Induction ◽  
In Vitro Propagation ◽  
Ornamental Plants ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Temperature Treatment ◽  
In Vitro Plant Regeneration

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)

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