A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses

被引：36

作者：

Wu, Faqiang ^{[1
]}

Hanzawa, Yoshie ^{[1
]}

机构：

[1] Calif State Univ Northridge, Dept Biol, Northridge, CA 91330 USA

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 131期

基金：

美国国家科学基金会;

关键词：

Cellular Biology; Issue; 131; Protoplast; soybean; Glycine max; unifoliate; transient gene expression; protein localization; SUBCELLULAR-LOCALIZATION; MESOPHYLL PROTOPLASTS; TRANSCRIPTION FACTORS; PLANT-PROTOPLASTS; TRANSFORMATION; BIOSYNTHESIS; PROMOTER; SYSTEM; MICROINJECTION; REGENERATION;

D O I：

10.3791/57258

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Soybean (Glycine max (L.) Merr.) is an important crop species and has become a legume model for the studies of genetic and biochemical pathways. Therefore, it is important to establish an efficient transient gene expression system in soybean. Here, we report a simple protocol for the preparation of soybean protoplasts and its application for transient functional analyses. We found that young unifoliate leaves from soybean seedlings resulted in large quantities of high quality protoplasts. By optimizing a PEG-calcium-mediated transformation method, we achieved high transformation efficiency using soybean unifoliate protoplasts. This system provides an efficient and versatile model for examination of complex regulatory and signaling mechanisms in live soybean cells and may help to better understand diverse cellular, developmental and physiological processes of legumes.

引用

页数：7

共 40 条

[1] A highly efficient transient protoplast system for analyzing defence gene expression and protein-protein interactions in rice [J].

Chen, Songbiao ;

Tao, Lizen ;

Zeng, Lirong ;

Vega-Sanchez, Miguel E. ;

Umemura, Kenji ;

Wang, Guo-Liang .

MOLECULAR PLANT PATHOLOGY, 2006, 7 (05) :417-427

[2] MAIZE POLYUBIQUITIN GENES - STRUCTURE, THERMAL PERTURBATION OF EXPRESSION AND TRANSCRIPT SPLICING, AND PROMOTER ACTIVITY FOLLOWING TRANSFER TO PROTOPLASTS BY ELECTROPORATION [J].

CHRISTENSEN, AH ;

SHARROCK, RA ;

QUAIL, PH .

PLANT MOLECULAR BIOLOGY, 1992, 18 (04) :675-689

[3] METHOD FOR THE ISOLATION OF PLANT PROTOPLASTS AND VACUOLES [J].

COCKING, EC .

NATURE, 1960, 187 (4741) :962-963

[4] INTEGRATION OF FOREIGN DNA FOLLOWING MICROINJECTION OF TOBACCO MESOPHYLL PROTOPLASTS [J].

CROSSWAY, A ;

OAKES, JV ;

IRVINE, JM ;

WARD, B ;

KNAUF, VC ;

SHEWMAKER, CK .

MOLECULAR & GENERAL GENETICS, 1986, 202 (02) :179-185

[5] Plant protoplasts: status and biotechnological perspectives [J].

Davey, MR ;

Anthony, P ;

Power, JB ;

Lowe, KC .

BIOTECHNOLOGY ADVANCES, 2005, 23 (02) :131-171

[6] GLUTATHIONE AND FUNGAL ELICITOR REGULATION OF A PLANT DEFENSE GENE PROMOTER IN ELECTROPORATED PROTOPLASTS [J].

DRON, M ;

CLOUSE, SD ;

DIXON, RA ;

LAWTON, MA ;

LAMB, CJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (18) :6738-6742

[7] The NAC domain-containing protein, GmNAC6, is a downstream component of the ER stress- and osmotic stress-induced NRP-mediated cell-death signaling pathway [J].

Faria, Jerusa A. Q. A. ;

Reis, Pedro A. B. ;

Reis, Marco T. B. ;

Rosado, Gustavo L. ;

Pinheiro, Guilherme L. ;

Mendes, Giselle C. ;

Fontes, Elizabeth P. B. .

BMC PLANT BIOLOGY, 2011, 11

[8] EXPRESSION OF GENES TRANSFERRED INTO MONOCOT AND DICOT PLANT-CELLS BY ELECTROPORATION [J].

FROMM, M ;

TAYLOR, LP ;

WALBOT, V .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (17) :5824-5828

[9] Evaluation of constitutive viral promoters in transgenic soybean roots and nodules [J].

Govindarajulu, Manjula ;

Elmore, James M. ;

Fester, Thomas ;

Taylor, Christopher G. .

MOLECULAR PLANT-MICROBE INTERACTIONS, 2008, 21 (08) :1027-1035

[10] INVITRO CULTURE OF WHITE CLOVER - CALLUS, SUSPENSION, PROTOPLAST CULTURE, AND PLANT-REGENERATION [J].

GRESSHOFF, PM .

BOTANICAL GAZETTE, 1980, 141 (02) :157-164

← 1 2 3 4 →