projects are exceedingly valuable tools for
genetic and physiological studies and/or synteny
analysis of leguminous plants. These resources
have been deposited with our resource center and
are available from“LegumeBase”for researchers
(Table22.1).Fig. 22.5 Root cloning, somatic embryogenesis, plant
regeneration, protoplast culture, transformation, and FOX
lines in superroots (super-growing root cultures) ofLotus
corniculatus(bird’s-foot trefoil) (Akashi et al. 1998 , 2003 ;
Tanaka et al. 2008 ; Himuro et al. 2011 ).aa 125-mL
Erlenmeyerflask containing a superroot culture displaying
typical root density at the end of a subculture period;broot
cultures derived from 10 lateral roots 28 days after
subculture (transferred to a 9-cm Petri plate for photog-
raphy);c,dsomatic embryogenesis from cultured roots;
eshoots, formed at wound sites of superroots, grow
vigorously when placed on a lighted shelf;fenzymatically
separated root tips after two hours of enzyme treatment;
gmicrocallus formation from isolated root protoplasts;
hmicrocolonies formed after 4 weeks of culture in agarose
disks in a 6-cm Petri plate;ishoot formation and shoot
elongation on protoplast-derived calli in a 9-cm Petri plate;
jresistant callus on MS medium containing 100 mg/L
kanamycin;k,lGUS activity in root (k) and leaf (l) tissues
of regenerated plants;mPetri plate with shoot-producing
callus derived from a leaf explant after 8 weeks;na
regenerated plant from superroots,oArabidopsisFOX-
superroots (numbers), wild type (WT) and superroots (SR)
ofLotus corniculatus. Shoots of FOX lines were rooted in
flat-bottomed test tubes. The roots had to penetrate 3 cm of
agar before coiling up at the bottom. Images of the test-
tube bottom were taken with a photocopier after 4 weeks
of culture252 M. Hashiguchi and R. Akashi