Development of an efficient Agrobacterium rhizogenes-mediated gene transformation system for millets

2021 ◽  
Author(s):  
Fathima Nashath
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Gene Transformation ◽  
Transformation System

scholarly journals Agrobacterium rhizogenes-mediated hairy root transformation as an efficient system for gene function analysis in Litchi chinensis

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Yaqi Qin ◽  
Dan Wang ◽  
Jiaxin Fu ◽  
Zhike Zhang ◽  
Yonghua Qin ◽  
...  
Keyword(s):  
Functional Genomics ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Gene Function ◽  
Function Analysis ◽  
Transformation System ◽  
Litchi Chinensis ◽  
Gene Function Analysis ◽  
Stem Segments

Abstract Background Litchi chinensis Sonn. is an economically important fruit tree in tropical and subtropical regions. However, litchi functional genomics is severely hindered due to its recalcitrance to regeneration and stable transformation. Agrobacterium rhizogenes-mediated hairy root transgenic system provide an alternative to study functional genomics in woody plants. However, the hairy root transgenic system has not been established in litchi. Results In this study, we report a rapid and highly efficient A. rhizogenes-mediated co-transformation system in L. chinensis using Green Fluorescent Protein (GFP) gene as a marker. Both leaf discs and stem segments of L. chinensis cv. ‘Fenhongguiwei’ seedlings were able to induce transgenic hairy roots. The optimal procedure involved the use of stem segments as explants, infection by A. rhizogenes strain MSU440 at optical density (OD600) of 0.7 for 10 min and co-cultivation for 3 days, with a co-transformation efficiency of 9.33%. Furthermore, the hairy root transgenic system was successfully used to validate the function of the key anthocyanin regulatory gene LcMYB1 in litchi. Over-expression of LcMYB1 produced red hairy roots, which accumulated higher contents of anthocyanins, proanthocyanins, and flavonols. Additionally, the genes involving in the flavonoid pathway were strongly activated in the red hairy roots. Conclusion We first established a rapid and efficient transformation system for the study of gene function in hairy roots of litchi using A. rhizogenes strain MSU440 by optimizing parameters. This hairy root transgenic system was effective for gene function analysis in litchi using the key anthocyanin regulator gene LcMYB1 as an example.

Attempt to Establish Direct Gene Transformation System to Seeds of Sweet Potato (Ipomoea batatas) Using Electroporation Method

2017 ◽  
Vol 19 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Lanzhuang Chen ◽  
Shigeru Masuoka ◽  
Yoshiko Nishimura ◽  
Tetsufumi Sakai ◽  
Yasuhiro Takahata ◽  
...  
Keyword(s):  
Sweet Potato ◽  
Ipomoea Batatas ◽  
Gene Transformation ◽  
Transformation System ◽  
Direct Gene

scholarly journals Study on obtaining of hairy root of Panax vietnamensis by using rol gene transformation via Agrobacterium rhizogenes

2014 ◽  
Vol 36 (1se) ◽  
Author(s):  
Ha Thi Loan ◽  
Duong Hoa Xo ◽  
Nguyen Quoc Binh ◽  
Nguyen Hoang Quan ◽  
Vu Thi Dao ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Gene Transformation ◽  
Rol Gene

Development of an Agrobacterium-mediated Transformation System for `Beurre Bosc' Pear

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan
Keyword(s):  
Shoot Proliferation ◽  
Induction Medium ◽  
Shoot Induction ◽  
Transformation System ◽  
Gus Histochemical Assay ◽  
Young Leaves ◽  
Leaf Tissues ◽  
Shoot Induction Medium ◽  
Basal Salts

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.

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