The Lotus japonicus Genome

betweenL. japonicusandL. tenuisby sequenc-
ing 17 full lengthL. tenuiscDNA clones isolated
by INIA/UACH in Chile. These cDNA clones
represent genes that are differentially expressed
during drought stress ofL. tenuisroot nodules
(Tapia et al. 2010 ; Tapia et al. unpublished). The
sequence identity between the coding regions of
these genes inL. tenuisandL. japonicusis above
95 %, showing how closely theseLotusspecies
are related. Again, this emphasises that the
genetic information obtained in the model spe-
ciesL. japonicusis of use in otherLotusspecies
such asL. tenuis.
Many of the microsatellite markers developed
fromL. japonicusGifu xL. japonicusMG-20
have been tested on DNA ofLotus corniculatus,
L. tenuis, Lotus uliginosusandLotus subbiflorus
using the following PCR conditions: 94° for

3 min followed by 40 rounds of 94°for 15 s, 54°

for 20 s and 72°for 30 s, andfinally 72°for 7 min

before cooling (Fig.3.3). In many cases, products

are amplified from these species. Such molecular

markers are potentially useful for breeding with

theLotusspecies used in agriculture. The map

positions inL. japonicusare already known for

these microsatellite markers (Sato et al. 2008 ).

In total, several hundred microsatellite mark-

ers have been developed by Kazusa DNA

Research Institute and mapped to a specific

position on theLotusgenome.

The cytogenetic map has been developed for

L. uliginosus(Ferreira et al. 2012 ). It shows a

high degree of macrosynteny toL. japonicus,but

it was interrupted by a translocation involving

chromosomes III and V, a new rearrangement for

the genus.

Table 3.2 Gene isolation/map-based cloning projects

Genes References Genes References

ArpC1 Hossain et al. ( 2012 ) Nfr5 Madsen et al. ( 2003 )

Astray Nishimura et al. (2002b) Nin Schauser et al. ( 1999 )

Brush Maekawa-Yoshikawa et al. ( 2009 ) Npl Xie et al. ( 2012 )

Castor Imaizumi-Anraku et al. ( 2005 ) Nsp1and

Nsp2

Heckmann et al. ( 2006 ), Murakami

et al. ( 2006 )

CCaMK/

snf1

Tirichine et al. (2006a,b) Nup133 Kanamori et al. ( 2006 )

Cerberus Yano et al. ( 2009 ) Nup85 Saito et al. ( 2007 )

CLE-RS2 Okamoto et al. ( 2009 ) PhyB Suzuki et al. ( 2011 )

Clv2 Krusell et al. ( 2011 ) Plenty Yoshida et al. ( 2010 )

Cyclops Yano et al. ( 2008 ) Pollux Imaizumi-Anraku et al. ( 2005 )

Fen1 Hakoyama et al. ( 2009 ) Rhl1 Karas et al. ( 2009 )

Gln2 Orea et al. ( 2002 ), García-Calderón

et al. ( 2012 )

Sen1 Hakoyama et al. (2012a)

Har1 Krusell et al. ( 2002 ), Nishimura et al.

(2002a)

snf2/hit1 Tirichine et al. ( 2007 ), Murray et al.

( 2007 )

Ign1 Kumagai et al. ( 2007 ) snf4 Tirichine et al. (2006a,b)

ITD4 Lombardo et al. ( 2006 ) Srh1and

sym105

Karas et al. ( 2005 ), Hossain et al.

( 2006 )

Klavier Miyazawa et al. ( 2010 ) SymRK Stracke et al. ( 2002 )

Lot1 Ooki et al. ( 2005 ) Sst1 Krusell et al. ( 2005 )

Napand

Pir

Yokota et al. ( 2009 ) Syp71 Hakoyama et al. (2012b)

NENA Groth et al. ( 2010 ) Tml Takahara et al. ( 2013 )

Nfr1 Radutoiu et al. ( 2003 ) Vrh1 Karas et al. ( 2005 )

26 N. Sandal and S. Sato