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Genomics and Applied Biology
,
2012, Vol.3 No.3, 1
-
7
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Research Report Open Access
The Acquisition and Expression Analysis of Polygalacturonase Gene
MF6
Related
to Fertility in
Brassica napus
L.
Rongxian Huang , Zhikang Tang , Shixing Guo , Yingze Niu
Rapeseed Research Center, Sichuan Agricultural University, Chengdu, 611130, P.R.China
Corresponding author,
niuyz01@126.com;
Authors
Genomics and Applied Biology 2012, Vol.3 No.1 doi: 10.5376/gab.2012.03.0001
Received: 25 May, 2012
Accepted: 28 Jun., 2012
Published: 30 Jun., 2012
Copyright
© 2012 Huang et al., This article was first published in Molecular Plant Breeding (2012, 10(3): 298-304) 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
Huang et al., 2012, The Acquisition and Expression Analysis of Polygalacturonase Gene
MF6
Related to Fertility in
Brassica napus
L., Genomics and Applied
Biology, 2012, Vol.3No.1 1-7 (doi: 10.3969/gab.2012.03.0001)
Abstract
The purpose of this study is to analyze the differentially expressed genes in the buds of male sterile plants which are
induced with a male sterilizing chemical “Hua-Sha-Ling WP1” in
Brassica napus
L., R121. Firstly, we isolated the differentially
expressed gene fragments from the male sterile buds through the suppression subtractive hybridization (SSH) and the reverse
northern dot blotting. Then, homology comparisons and functional analysis were conducted through online database NCBI. Finally,
the differentially expressed gene was further analyzed through reverse transcription quantitative PCR. In the results, we obtained a
gene fragment which was 100% homologous to the polygalacturonase gene (
MF6
) in Arabidopsis from the 6 positive clones. It is
reported that the polygalacturonase gene (
MF6
) is closely related to the development and maturation of pollen. The amount of
MF6
gene expression in flower buds with different lengths and different fertilities was analyzed quantitatively. The results showed that the
expression of
MF6
gene in the male fertile flower buds was higher than that in the male sterile flower buds. And the differences are
about 18.53 times and 43.15 times when the bud lengths are 1.00~1.50 mm and 3.5~4.0 mm, respectively. Therefore, “Hua-Sha-Ling
WP1” may inhibit the expression of the
MF6
gene during the meiosis stages of pollen mother cell and cause the abortion of pollens.
At the same time, the
MF6
gene expression may also be associated with the elongation of filaments.
Keywords
Brassica napus
L.; Chemical male sterilization; Differentially expressed genes;
MF6
; Pollen abortion
Background
Chemical hybridization has become one of the most
important ways of utilization of heterosis in crops
such as rapeseed, with advantages of thorough male
sterilization, freedom in parental selection and
combination, freedom from artificial emasculation and
so on (Zhang et al., 2011). It can also shorten the breeding
period of hybrid cultivars and raise the breeding efficiency
(Zhang et al., 2011). Since Naylor (1950) reported that
Maleic Hydrazide (MH) could inhibit the development
of anther, resulting in male sterility in plants in the
1950s, there have been a large number of scientists who
studied the selection, the effects and the mechanism of
chemical hybridizing agent (CHA). For example, in
the process of male sterility induction by CHA in
rapeseed, the release rate of ethylene was increased,
the soluble protein content was decreased, the total
free amino acid contents were decreased, the free proline
content in anthers was decreased, and the enzyme
activity was altered (Fan et al., 2008). However, reports
on molecular mechanism of the male sterilization by
CHA were rare. Studies on the effects of CHA on the
expression of relevant genes and the molecular mechanism
of plant male sterility will help us with the selection,
the synthesis of chemical hybridizing agent in a cheaper
cost, a higher efficiency and lower pollution.
At the same time, we may also transfer the genes
related to male sterilization into superior parents to
fasten the breeding progress. In this study, we used an
effective CHA, called “Hua-Sha-Ling WP1”, to induce
male sterility in a rapeseed line, R121. We used the
suppression subtractive hybridization method (SSH) and
the reverse northern hybridization technology to
display the differentially expressed gene in the flower
buds. Homology comparisons and functional analyses
were conducted with the identified fragments of genes
based on the internet data. The mechanism of male
sterility induced by “Hua-sha-ling WP1” was discussed
according to the quantitative expressions of the
relevant gene.
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