HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text Free Full Text (PDF) Free Alert me when this article is cited Alert me if a correction is posted Services Related articles in Crop Science Similar articles in this journal Similar articles in ISI Web of Science Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (4) Citing Articles via Google Scholar Google Scholar Articles by Yashitola, J. Articles by Sonti, R. V. Search for Related Content PubMed Articles by Yashitola, J. Articles by Sonti, R. V. Agricola Articles by Yashitola, J. Articles by Sonti, R. V. Related Collections Rice Cell Biology & Molecular Genetics Seed Production

A Sequence Specific PCR Marker for Distinguishing Rice Lines on the Basis of Wild Abortive Cytoplasm from Their Cognate Maintainer Lines

^a Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500007, India
^b Directorate of Rice Research, Rajendranagar, Hyderabad-500030, India

View larger version (38K): [in a new window]   Fig. 1. PCR amplification of a DNA sequence specific to CMS lines of rice. PCR was performed with RM9 microsatellite primers. Lane 1 contains a DNA molecular weight marker ( DNA digested by HindIII); Lane 2 contains PCR amplified product of a maintainer line (IR58025B); Lane 3, the hybrid (DRR H1), and Lane 4, CMS line (IR58025A). An extra DNA band, which is only present in the hybrid and CMS line but absent in the maintainer line, is indicated by the arrow.

View larger version (19K): [in a new window]   Fig. 2. Nucleotide sequence of a DNA fragment amplified from WA-CMS lines of rice. A 323-bp long CMS specific DNA fragment that was amplified with RM9 primers was purified and sequenced as described in Materials and Methods. A segment of this sequence, extending from nucleotides 37-319, exhibits 97% identity to a region of rice mitochondrial DNA (DBJ accession #D21251). Nucleotides 1 to 36 and 319 to 323 of this sequence are not homologous to the sequence in D21251. The unique sequences are indicated in bold letters. Arrows indicate the binding sites of the RM9 primers.

View larger version (49K): [in a new window]   Fig. 3. A PCR assay using sequence specific primers to distinguish WA-CMS and maintainer lines of rice. PCR was performed with cms primers. Lane 1: kilobase DNA marker; Lane 2: IR58025A (CMS); Lane 3: DRRH1 (hybrid); Lane 4: IR58025B (maintainer). PCR product was obtained only when template DNA is from the CMS line and hybrid.

View larger version (33K): [in a new window]   Fig. 4. A multiplex PCR assay for distinguishing CMS and maintainer lines of rice. PCR was performed by multiplexing cms primers with RG136 primers. M indicates a kilobase DNA ladder as a molecular weight marker. Lanes 1 and 14: IR58025A and B; Lanes 2 and 15: IR62829A and B; Lanes 3 and 16: IR68888A and B; Lanes 4 and 17: IR68897A and B; Lanes 5 and 18: CRMS31A and B; Lanes 6 and 19: DRR2A and B; Lanes 7 and 20: Pusa5A and B; Lanes 8 and 21: PMS8A and B; Lanes 9 and 22: PMS10A and B; Lanes 10 and 23: PMS11A and B; Lanes 11 and 24: PMS12A and B; Lanes 12 and 25: DRR3A and B; Lanes 13 and 26: IR68886A and B.

View larger version (42K): [in a new window]   Fig. 5. Purity analysis of seed stock of CMS line. Genomic DNA was isolated individually from 108 plants and PCR was performed by multiplexing cms primers with RG136 primers. Data from a representative set of 11 plants are presented. The arrow indicates CMS-specific fragment. The asterisk indicates plants determined to be maintainer lines.