Identification of cadmium-induced genes in maize seedlings by suppression subtractive hybridization

doi:10.1007/s11783-010-0250-x

Front Envir Sci Eng Chin

2010, Vol. 4

Issue (4) : 449-458 https://doi.org/10.1007/s11783-010-0250-x

RESEARCH ARTICLE

Identification of cadmium-induced genes in maize seedlings by suppression subtractive hybridization

Quanlin DAI¹, Baifei HUANG^1,3, Zhongyi YANG¹(

), Jiangang YUAN¹(

), Junzhi YANG²

1. State Key Laboratory of Biocontrol, Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; 2. College of Natural Resources, University of California, Berkeley, CA 94720, USA; 3. Department of Safety and Environmental Engineering, Hunan Institute of Technology, Hengyang 421002, China

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Abstract

A maize variety, Huatian-1, had an unusually low translocation rate of cadmium (Cd) (59.6 mg·kg^-1 in the roots and 0.093 mg·kg^-1 in the grain) compared to 24 other varieties while being grown in soils with 16.50 mg·kg^-1 Cd. This indicates that this particular species may have special mechanisms that affect the absorption and translocation pattern of Cd. In this paper, the technique of suppression subtractive hybridization (SSH) was used to isolate and identify Cd-induced genes from Huatian-1 hydroponically ?exposed? to? 0.1 mM ?CdCl₂ ?for? 1 h,? 12 h, 24 h, and 48 h. We found a total of 15 differentially expressed genes in the four groups; 2, 3, 4, and 6 genes were from the groups of 1 h, 12 h, 24 h, and 48 h treatment, respectively. Phospholipase PLDb1 mRNA, adenosine triphosphate (ATP) phosphoribosyl transferase 2, and Sp17 were turned on in the maize in response to Cd stress, and it might provide new clues to explain the mechanism of maize tolerance to Cd.

Keywords Zea mays L. cadmium (Cd) suppression subtractive hybridization

Corresponding Author(s): YANG Zhongyi,Email:adsyzy@mail.sysu.edu.cn; YUAN Jiangang,Email:yuanjg@mail.sysu.edu.cn

Issue Date: 05 December 2010

Cite this article:

Quanlin DAI,Baifei HUANG,Zhongyi YANG, et al. Identification of cadmium-induced genes in maize seedlings by suppression subtractive hybridization[J]. Front Envir Sci Eng Chin, 2010, 4(4): 449-458.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-010-0250-x
https://academic.hep.com.cn/fese/EN/Y2010/V4/I4/449

Fig.1

Results of the differential screening of 48 h treatment cDNA library

Note: (a) Dot blots hybridized with cDNA probes made from forward unsubtracted cDNA (Cd²⁺-treated cDNA un-subtracted probes).

(b) Dot blots hybridized with cDNA probes made from forward subtracted cDNA (Cd²⁺-treated cDNA with no-treatment cDNA subtracted probes).

(d) Dot blots hybridized with cDNA probes made from reverse subtracted cDNA (no-treatment cDNA with Cd²⁺-treated cDNA subtracted probes).

? Clones hybridized to the forward subtracted/unsubtracted tester probes but not to the reverse subtracted/unsubtracted driver probes.

? Clones hybridized only to the forward subtracted probes.

? Clones hybridized to four probes, but the signals of forward subtracted and unsubtracted probes are more intensive than that of the reverse subtracted or unsubtracted probes.

◇ Negative controls.

Fig.2

Virtual Northern blot analysis of partial cDNA fragments of Library IV (T: Tester, cDNA of Cd-treated tissue; D: Driver, cDNA of no-treatment tissue)

Tab.1

Results of sequences for the putative cDNAs clones from four libraries induced by Cd isolated by suppression subtractive hybridization

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