Resistance gene analogues from rice: cloning, sequencing and mapping |
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Authors: | R Mago S Nair M Mohan |
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Institution: | (1) International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110 067, India Fax:+91 11 618 7680 E-mail: mohanm@del2.vsnl.net.in, IN |
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Abstract: | Degenerate oligonucleotide primers were designed on the basis of nucleotide-binding-site (NBS) motifs conserved between resistance
genes of Arabidopsis, flax and tobacco and subsequently used as PCR primers to amplify resistance gene analogues (RGA) in rice. Primers amplified
a major band of approximately 500 bp. Restriction analysis of the amplified product revealed that the band was made up of
several different fragments. Many of these fragments were cloned. Sixty different cloned fragments were analysed and assigned
to 14 categories based on Southern blot analysis. Fourteen clones, each representing one of the 14 categories of RGAs were
mapped onto the rice genetic map using a Nipponbare ( japonica)בKasalath’ (indica) mapping population consisting of 186 F2 lines. Of the 14 clones representing each class 12 could be mapped onto five different chromosomes of rice with a major cluster
of 8 RGAs on chromosome 11. Our results indicate that it is possible to use sequence homology from conserved motifs of known
resistance genes to amplify candidate resistance genes from diverse plant taxa.
Received: 23 September 1998 / Accepted: 28 November 1998 |
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Keywords: | Degenerate oligonucleotides Disease resistance Leucine-rich repeats (LRR) Nucleotide-binding site (NBS) Resistance gene analogues (RGA) |
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