Abstract
Nodule samples from 90A. nepalensis individuals were collected at five sites in the Hengduan Mountains. PCR-RFLP analysis of IGS betweennifD andnifK genes was directly applied to unculturedFrankia strains in the nodules. Sizes of thenifD-nifK IGS amplicons and genetic distance between the RFLP patterns from these samples were noticeably different, indicating significant genetic variation in theFrankia population. There were some nodule samples, which produced more than one PCR fragment, and compound RFLP patterns, indicating thatFrankia strains with different PCR-RFLP patterns coexisted in the same host plant under natural conditions. Among the 29 restriction patterns obtained, 5 patterns were found in more than one population and occurred in the majority of samples, while each of the other 24 patterns were represented by only one or two samples and were endemic to a particular population. From the calculatedGst and UPGMA cluster analysis, genetic diversity ofFrankia strains was inferred to be related to climate and glaciation history in the Hengduan Mountains.
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References
Ager A A, Heilman P E and Stettler R F 1993 Genetic variation in red alder (Alnus rubra) is relation to native climate and geography. Can. J. For. Res. 23, 1930–1939.
Buntjer J B 2001 PhylTools (Phylogenetic Computer Tools) version 1.32. Laboratory of Plant Breeding, Wageningen Agriculture University, Netherlands.
Chen Z D 1999 Origin and dispersal of Betulaceae.In The Geography of Spermatophytic Families and Genera. Ed. A M Lu. pp. 236–258. Science Press, Beijing.
Chen Z D, Manchester S R and Sun H Y 1999 Phylogeny and evolution of the Betulaceae as inferred from DNA sequences, morphology, and paleobotany. Am. J. Bot. 86, 1168–1181.
Excoffier L and Smouset P E 1994 Using allele frequencies and geographic subdivision to reconstruct gene trees within a species: Molecular variance parsimony. Genetics 136, 343–359.
Hahn D, Nickel A and Dawson J 1999 AssessingFrankia populations in plants and soil using molecular methods. FEMS Microbiol. Ecol. 29, 215–227.
King A and Ferris C 1998 Chloroplast DNA phylogeography ofAlnus glutinosa (L.) Gaertn. Mol. Ecol. 7, 1151–1161.
Leitz G, Lundberg C, Fällman E, Axner O and Sellstedt A 2003 Laser-based micromanipulation for separation and identification of individualFrankia vesicles. FEMS Microbiol. Lett. 224, 97–100.
Li H, Guo H J and Dao Z L 2000 Flora of Gaoligong Mountains. Science Press, Beijing.
Lumini E and Bosco M 1999 Polymerase chain reaction - restriction fragment length polymorphisms for assessing and increasing biodiversity ofFrankia culture collections. Can. J. Bot. 77, 1261–1269.
Mitchell L G, Bodenteich A and Merril C R 2002 Silver staining of DNA.In Basic DNA and RNA Protocols. Ed. A J Harwood. pp. 67–71. Science Press, Beijing.
Murry M, Konopka A, Pratt D and Vandergon T 1997 The use of PCR-based typing methods to assess the biodiversity ofFrankia nodule endophytes of the actinorhizal shrub Ceanothus. Physiol. Plantarum 99, 714–721.
Nalin R, Normand P and Domenach A M 1997 Distribution and N2-fixing activity ofFrankia strains in relation to soil depth. Physiol. Plantarum 99, 732–738.
Ritchie N J and Myrold D D 1999 Phylogenetic placement of unculturedCeanothus microsymbionts using 16S rRNA gene sequences. Can. J. Bot. 77, 1208–1213.
Rouvier C, Prin Y, Reddell P, Normand P and Simonet P 1996 Genetic diversity amongFrankia strains nodulating members of the family Casuarinaceae in Australia revealed by PCR and restriction fragment length polymorphism analysis with crushed root nodules. Appl. Environ. Microb. 62, 979–985.
Sarma G and Misra A K 2002 Mixed genotypes in some axenic cultures ofFrankia. Indian J. Microb. 42, 11–14.
Simonet P, Navarro E, Rouvier C, Reddell P, Zimpfer J, Dommergues Y, Bardin R, Combarro P, Hamelin J, Domenach A, Gourbière F, Prin Y, Dawson J and Normand P 1999 Co-evolution betweenFrankia populations and host plants in the family Casuarinaceae and consequent patterns of global dispersal. Environ. Microb. 1, 525–533.
Soltis D E, Gitzendanner M A, Strenge D D and Soltis P S 1997 Chloroplast DNA intraspecific phylogeography of plants from the Pacific Northwest of North America. Pl. Syst. Evol. 206, 353–373.
Varghese R, Chauhan V S and Misra A K 2003 A hypervariable spacer regions are good sites for developing specific PCR-RFLP markers and PCR primers for screening actinorhizal symbionts. J. Biosci. 28, 437–442.
Verghese S K and Misra A K 2000 PCR-RFLP based screening ofFrankia genotypes in alder nodules having different levels of nitrogenase activity. Symbiosis 28, 337–350.
Wu S H, Zhang H W and Xiong Z 2001 Biodiversity ofFrankia strains in nodules fromAlnus andHippophae by ARDRA. Chin. J. Appl. Ecol. 12, 883–886.
Xiong Z, Li W J, Zhang Z Z and Jiang C L 2001 The influence of altitude on the genetic of microsymbionts ofAlnus nepalensis-Frankia. Chin. J. Southwest Forest. Col. 21, 205–209.
Xu J R and Jiang H J 1984 Forests in Yunnan Province. Yunnan Science and Technology Press, Kunming.
Yeh F C, Yang R C, Royle T B J, Ye Z H and Mao J X 1997 POPGENE V1.31. http://www.ualberta.ca/-fyeh
Ying J S 2001 Species diversity and distribution pattern of seed plants in China. Chin. Biodivers. Sci. 9, 393–398.
Zhang R Z, Zheng D, Yang Q Y and Liu Y H 1997 Physical geography of Hengduan Mountains. Science Press, Beijing.
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Dai, Y.M., He, X.Y., Zhang, C.G. et al. Characterization of genetic diversity ofFrankia strains in nodules ofAlnus nepalensis (D. Don) from the Hengduan Mountains on the basis of PCR-RFLP analysis of thenifD-nifK IGS. Plant Soil 267, 207–212 (2004). https://doi.org/10.1007/s11104-005-5028-8
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DOI: https://doi.org/10.1007/s11104-005-5028-8