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Legume Genomics and Genetics (online), 2011, Vol. 2, No.3, 14-21
http://lgg.sophiapublisher.com
14
Research Article Open Access
Heat Shock Factor 8 Introducing into Soybean (
Glycine max
) by
Agrobacterium
-M-
ediated Transformation
Xiaofei Tang
1
, Lijun Liu
1
, Mingjie Gao
1
, Zhe Yang
1
, Guofeng Pu
1
, Lei Zhang
1
, Lai Wei
2
1. Soybean Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin, 150086, P.R. China
2. Northeast Agricultural University, Harbin, 150030, P.R. China
Corresponding author email: nkyssbd@126.com;
Authors
Legume Genomics and Genetics 2011, Vol.2 No.3 DOI:10.5376/lgg.2011.02.0003
Received: 13 Oct., 201
Accepted: 12 Dec., 2011
Published: 16 Dec., 2011
This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
Preferred citation for this article as:
Tang et al., 2009,
Heat Shock Factor 8 Introducing into Soybean (
Glycine max
) by
Agrobacterium
-Mediated Transformation, Molecular Plant Breeding, 7(3):
444-450 (doi: 10.3969/mpb.007.000444)
Abstract
Environmental stress seriously affects growth and development of crops. Heat shock transcription factor (heat shock
factor 8, HSF8) is a kind of proteins that play important roles in the reactions of heat shock, the main functions of heat shock gene
are binding the corresponding heat shock elements in the process of expression, initiating the gene transcription process and,
ultimately, facilitating the expression of heat shock protein (HSP) gene. In this paper, we inserted
hsf8
gene into dicotyledonous
expression vector pCAMBIA3300 that contains the selection marker,
bar
gene. The construct, named pCAMBIA3300-HSF8, has
been transferred into new soybean lines, Hajiao5337 and Hajiao5489, mediated by
Agrobacterium tumefaciens
. In the practice of
soybean genetic transformation, we explored the impact factors on the transformation of soybean cotyledonary-node mediated by
Agrobacterium
, and optimized the transformation conditions. It is the way to increase the selection efficiency by delaying the
screening after co-culture with optimal concentration of glufosinate-ammonium 3.5 mg/L in selection culture medium. We obtained
the T
1
generation transgenic plants with pCAMBIA3300-HSF8 by
Agrobacterium
-mediated approach derived from Hajiao5337 and
Hajiao5489, of which 17 plants were proved by PCR to be positive transgenic plants. Furthermore, transcription level of
hsf8
gene in
T
1
transgenic plants with glufosinate-ammonium resistance was detected by Real-time PCR method, the results showed that 9 of 17
plants had much higher expression level than that of the reference of Hajiao5337, whereas one transgenic plant had lower level than
that of another reference Hajiao5489.
Keywords
Soybean (
Glycine max
); Heat shock factor 8 gene (
hsf8
);
Agrobacterium
-mediated transformation; Heat tolerance
Background
Soybean is the world's most important grain and oil
crops. Total annual production of soybean in China
could not meet the increasing domestic consumer’s
demand, which would contribute to the low yield of
soybean. Increasing the level of soybean yield
depends on the soybean varieties with the resistant
abilities to adverse growing conditions and with the
effective use of nutrients. Environmental stresses such
as drought, soil salinity and extreme temperatures and
so on are serious impact on crop growth and
development, resulting in yield decrease (Wang et al.,
2003), of which the adverse temperature is dominating
factor. In fact, Soybean is a kind of crop lack of high
temperature tolerance, the temperature is too high or
too dry at the flowering and pod developing stage that
will lead to soybean yield reducing. It was found that
synthesis of heat shock proteins (heat stress / shock
protein, HSP) was positively correlated to acquiring
biological heat tolerance. HSP synthesis can improve
the response ability of organisms, particularly in the
capacity of heat tolerance (Zhu et al., 2006).It is said
that many organisms receiving heat shock treatment
under the below of the lethal temperature could
improve the survival opportunity in the face of
adversity.
Expression and regulation of Heat shock protein gene
may be achieved by heat shock transcription factor
(HSF), which mainly occurs at the transcriptional
level. In higher organisms, potential HSF might be a
potential activator in the heat shock environment,
which would be a process along with the intra- and
inter-molecular interactions of HSF as well as the
interaction between HSF and heat shock / HSE (heat