The polyembryo gene (OsPE) in rice

Anju Puri, P. Osman Basha, Mankesh Kumar, Deepak Rajpurohit, Gursharn S. Randhawa, Shahryar F. Kianian, Anantharama Rishi, Harcharan S. Dhaliwal

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

T-DNA insertional mutagenesis is one of the most important approaches for gene discovery and cloning. A fertile polyembryo mutant generated by T-DNA/Ds insertion in Oryza sativa, cv. Basmati 370 showed twin or triple seedlings at a frequency of 15-20%. T-DNA insertion was confirmed by 950 bp hpt gene amplification in the promoter region of the candidate gene. The annotated protein corresponding to the OsPE candidate gene has been reported as a hypothetical protein in O. sativa. OsPE gene lacked functional homologs in other species. No OsPE paralog was found in rice. No conserved domains were found in the protein coded by OsPE. RT-PCR showed the expression of OsPE gene in Basmati 370 shoots. Full-length OsPE gene was cloned in Basmati 370. The combined use of Southern blot, genome walking, TAIL-PCR, RT-PCR techniques, and bioinformatics led to the identification of a candidate gene controlling the multiple embryos in rice. There is gain of function, i.e., multiple embryos in the seeds in the knockout mutant OsPE whereas its wild-type allele strictly controls single embryo per seed. The seeds with multiple embryos are distributed at random in the rice mutant panicle. The origin of multiple embryos, whether apomictic, zygotic or both is under investigation.

Original languageEnglish (US)
Pages (from-to)359-366
Number of pages8
JournalFunctional and Integrative Genomics
Volume10
Issue number3
DOIs
StatePublished - Aug 2010
Externally publishedYes

Bibliographical note

Funding Information:
Acknowledgements We thank Dr. Kuldeep Singh, PAU Ludhiana for providing rice SSRs for BSA. We also thank Dr. K. S. Gill, WSU for his thoughtful review and suggestions in improving this article. The financial assistance by CSIR (India) to A.B and NSF (USA) to S.K.F. and H. S. D. is highly acknowledged.

Keywords

  • Cloning
  • Insertional mutagenesis
  • Oryza sativa
  • Polyembryo
  • T-DNA
  • T-DNA/Ds

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