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Molecular Breeding

, Volume 32, Issue 4, pp 969–976 | Cite as

A comparative study of the atp9 gene between a cytoplasmic male sterile line and its maintainer line and further development of a molecular marker specific for male sterile cytoplasm in kenaf (Hibiscus cannabinus L.)

  • Yanhong Zhao
  • Peng Chen
  • Xiaofang Liao
  • Bujin Zhou
  • Jian Liao
  • Zhipeng Huang
  • Xiangjun Kong
  • Ruiyang ZhouEmail author
Article

Abstract

mtDNA was isolated from cytoplasmic male sterility (CMS) line P3A and its maintainer P3B of kenaf (Hibiscus cannabinus L.). The atp9 gene and its two flanking sequences were obtained using homology cloning and high-efficiency thermal asymmetric interlaced PCR methods. The coding sequences showed only two base pairs difference between the CMS and its maintainer, and shared a homology of over 87 % with atp9 genes from other species in GenBank. However, when comparing the flanking sequences, a 47-bp deletion was characterized at the 3′ flanking sequence of atp9 in the CMS line. Quantitative PCR analysis indicated that the expression level of atp9 in the CMS line was 0.937-fold that of its maintainer. Furthermore, the respiratory rate of anthers in the CMS line was markedly lower than that of its maintainer. The results indicated that the 47-bp deletion at the 3′ flanking sequence of atp9 and/or down-regulated expression of the atp9 gene in the CMS line might be closely related to CMS in kenaf. To confirm whether the 47-bp deletion was specific to cytoplasm of male sterile lines, another 21 varieties were used for further analysis. The results showed that the 47-bp deletion was specific to male sterile cytoplasm (MSC) of kenaf. Based on these, a specific molecular marker was developed to distinguish the MSC from male fertile cytoplasm of kenaf.

Keywords

Kenaf (Hibiscus cannabinus L.) Cytoplasmic male sterility (CMS) atp9 Molecular marker Quantitative PCR Respiratory rate 

Abbreviations

atp9

ATPase subunit 9 gene

CMS

Cytoplasmic male sterility

CTAB

Cetyltrimethyl ammonium bromide

GAPDH

Glyceraldehyde phosphate dehydrogenase gene

hiTAIL-PCR

High-efficiency thermal asymmetric interlaced PCR

MFC

Male fertile cytoplasm

MSC

Male sterility cytoplasm

mtDNA

Mitochondria DNA

ORF

Open reading frame

qPCR

Quantitative PCR

WT

Wild type

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 30900912, 31060199, 31171600).

Supplementary material

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Supplementary material 1 (TIFF 511 kb)
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Supplementary material 2 (TIFF 3497 kb)
11032_2013_9926_MOESM3_ESM.doc (38 kb)
Supplementary material 3 (DOC 38 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Yanhong Zhao
    • 1
    • 2
  • Peng Chen
    • 1
  • Xiaofang Liao
    • 1
  • Bujin Zhou
    • 1
  • Jian Liao
    • 1
  • Zhipeng Huang
    • 1
  • Xiangjun Kong
    • 1
  • Ruiyang Zhou
    • 1
    Email author
  1. 1.College of AgricultureGuangxi UniversityNanningChina
  2. 2.Cash Crop Institute of Guangxi Academy of Agricultural SciencesNanningChina

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