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Plant and Cell Physiology, 2002, Vol. 43, No. 11 1351-1358
© 2002 Oxford University Press

Expression of Symbiotic and Nonsymbiotic Globin Genes Responding to Microsymbionts on Lotus japonicus

Toshiki Uchiumi1,5, Yoshikazu Shimoda2, Tomoko Tsuruta2, Yumi Mukoyoshi1, Akihiro Suzuki1, Keishi Senoo3, Shusei Sato4, Tomohiko Kato4, Satoshi Tabata4, Shiro Higashi1 and Mikiko Abe1

1 Department of Chemistry and BioScience, Kagoshima University, Kagoshima, 890-0065 Japan
2 Graduate School of Science and Technology, Kagoshima University, Kagoshima, 890-0065 Japan
3 Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, 113-8657 Japan
4 Kazusa DNA Research Institute, Chiba, 292-0812 Japan

Leguminous plants have both symbiotic and nonsymbiotic hemoglobin (sym- and nonsym-Hb) genes. Three symbiotic (LjLb1, 2, 3) and one nonsymbiotic (LjNSG1) Hb genes were isolated from a genomic library of Lotus japonicus MG20 Miyakojima. Two motif sequences (AAAGAT and CTCTT) critical for nodule specific expression were conserved on the 5'-upstream sequence of LjLb1, 2 and 3. The 5'-upstream region of LjNSG1 contained the sequence consensus for nonsym-Hb. RT-PCR with specific primer sets for each LjLb gene showed that all the sym-Hb genes (LjLb1, 2, 3) were expressed specifically and strongly in root nodules. The expression of LjLb1, 2 and 3 could not be detected in root, leaf or stem of a mature plant, whereas low level expression was detected in seedlings by RT-PCR. This suggests that sym-Hbs may have another unknown function besides being oxygen transporter for the microsymbiont in symbiotic nitrogen fixation. The expression of LjNSG1, examined with RT-PCR, was detected at low level in root, leaf and stem. The expression of LjNSG1 was enhanced in root nodules, whereas it was repressed in roots colonized by mycorrhizal fungi Glomus sp. R10. The repression of the nonsym-Hb gene was also observed in the roots of Medicago sativa colonized by Glomus sp. R10.

5 Corresponding author: E-mail, uttan@sci.kagoshima-u.ac.jp; Fax, +81-99-285-8163.


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