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Molecular Plant Breeding 2013, Vol.4, No.30, 247
-
253
http://mpb.sophiapublisher.com
247
Research Report Open Access
Genetic Transformation of
Brassica napus
with
MSI-99m
Gene Increases
Resistance in Transgenic Plants to
Sclerotinia sclerotiorum
Xuelian Sang , Dengwei Jue , Liu Yang , Xiao Bai , Min Chen , Qing Yang
College of life sciences, Nanjing Agricultural University, Nanjing, 210095, P. R. China
Corresponding authors email: qyang19@njau.edu.cn
Authors
Molecular Plant Breeding, 2013, Vol.4, No.30 doi: 10.5376/mpb.2013.04.0030
Received: 29 Jul., 2013
Accepted: 26 Sep., 2013
Published: 27 Sep., 2013
Copyright
©2013 Sang et al. This is an open access article published under the terms of the 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:
Sang et al., Genetic Transformation of
Brassica napus
with
MSI-99m
Gene Increases Resistance in Transgenic Plants to
Sclerotinia sclerotiorum
, Molecular
Plant Breeding, Vol.4, No.30 247
-
253 (doi: 10.5376/mpb.2013.04.0030)
Abstract
Magainins are a class of antimicrobial peptides isolated from skin secretions of the African clawed frog
Xenopus laevis
.
MSI-99
is a synthesized magainin II analogue with inhibitory effects to a wide spectrum of microbial organisms, including bacteria,
fungus and viruses.
Sclerotinia sclerotiorum
is one of the most destructive pathogens in rape (
Brassica napus
), causing devastating
yield losses. To evaluate its resistance to rape
Sclerotina
rot, we transferred the
MSI-99m
gene (modified
MSI-99
) into Chinese rape
variety Zhongyou 821 using
Agrobacterium
-mediated method. Nine transformed lines carrying a
MSI-99m
expression vector were
detected by polymerase chain reaction (PCR), among which seven lines expressed
MSI-99m
gene according to qRT–PCR analysis.
Disease resistance analysis consistently showed that the high level expression of
MSI-99m
increased resistance to
S. sclerotiorum
in
transgenic rape lines. This result demonstrated that
MSI-99m
gene may be applied as a resistant gene resource in rape for the
improvement of rape varieties
.
Keywords
Brassica napus
;
MSI-99m
; Transformation;
Sclerotinia sclerotiorum
; Disease resistance
Introduction
Rape (
Brassica napus
) is one of the world’s major
oilseed crops and the most important source of edible
oil in China (Momoh et al., 2002). But rape is also
susceptible to a series of fungal diseases, such as
Sclerotinia
rot, downy mildew, and white rust. Of the
fungal diseases,
Sclerotinia
rot is the most destructive
disease which could cause the rotting of leaves, stems
and pods, resulting in a tremendous loss in seed yield
and quality in different regions all over the world
(Dong et al., 2008; Guo and Stotz, 2007). In China
this disease can cause a 15% yield loss nationally
every year. Biotechnology breeding is a valuable
alternative way for solving the problem above and has
been successfully applied in plant resistance
improvement.
In recent years, some successes in genetic transformation
for introducing resistant genes into rape have been
reported. The expression of antimicrobial peptide gene
PmAMP1
in rape conferred greater protection against
Alternaria brassicae
,
Leptosphaeria maculans
and
Sclerotinia sclerotiorum
(Verma et al., 2012).
Meanwhile, constitutively expressing
OXO
in
transgenic rape displayed considerably increased
OXO activity and enhanced resistance to
S.
sclerotiorum
(Dong et al., 2008). However, in
production, the appearance of new races of pathogens
caused by variation often results in loss of host
resistance, which could be made up through using
genes with a wide spectrum of resistance.
Antimicrobial peptides (AMP) are small cationic,
pore-forming peptides with inhibitory activities
against a spectrum of pathogens, which usually are
obtained from animals (Rao, 1995) or even plants
(Broekaert et al., 1997). Magainin II is one of the
earliest reported antimicrobial peptides isolated from
the skin secretions of the African clawed frog
Xenopus
laevis
(Zasloff, 1988), which has been studied
extensively because of its strong inhibitory effects on
a broad spectrum of plant pathogens (Aboudy et al.,
1994) and its potential application in disease
resistance breeding (O’Callaghan et al., 2008).
MSI-99
, a magainin II analogue, is synthesized by
making several modifications in magainin II (Everett,
1994; Maloy and Kari, 2004). The
gene has been