Development of a DNA marker for distinguishing CMS lines from fertile lines in rice (Oryza sativa L.) |
| |
Authors: | Nirmala Rajendran Ramkumar Gandhimani Sukhpal Singh Kadirvel Palchamy |
| |
Institution: | (1) Research and Training Centre, Barwale Foundation, No. 8-2-703, A.G. Heights, Road No. 12, Banjara Hills, Hyderabad, Andhra Pradesh, 500034, India |
| |
Abstract: | The main objective of the present study was to identify mitochondrial DNA based marker, which can distinguish male sterile
and fertile counterparts of the cytoplasmic male sterile (CMS) lines used in production of rice hybrids. Amplified fragment
length polymorphism (AFLP) analysis in CMS lines: IR58025A & IR62829A and their respective maintainers: IR58025B & IR62829B
identified a polymorphic DNA fragment of about 510 bp size that was present in both CMS (A) and absent in their maintainer
(B) lines. Sequencing followed by database analysis of the polymorphic fragment indicated about 97% similarity with mitochondrial
NADH gene subunits of rice, maize and wheat. Based on the variable sequence regions, a site specific primer pair (BF-STS-401)
was designed. PCR analysis showed that BF-STS-401 could amplify a strong band of 464 bp size in CMS and a faint band of the
same size in maintainer line. To act as a positive control and avoid possible errors in PCR, BF-STS-401 was multiplexed with
a new primer pair (BF-STS-402), derived from mitochondrial atp9 subunit of rice, producing monomorphic amplification indiscriminately in both CMS and maintainer lines. Both the primer pairs
in combination clearly differentiated CMS lines from their corresponding maintainer lines. This primer combination was validated
in a set of diverse genotypes consisting of different sources of CMS lines, restorer lines, hybrids, varieties and mixed samples
from private seed companies. Our results suggested that the multiplex primer pairs developed in this study can be effectively
utilized to assess the genetic purity in commercial seed lots of CMS lines and hybrids of rice. |
| |
Keywords: | AFLP CMS Genetic purity STS marker Rice |
本文献已被 SpringerLink 等数据库收录! |
|