New insights in conservation of Sophora toromiro (Phil.) Skottsb., emblematic species of the South Pacific Jaime Espejo and Francisco Rodriguez This฀paper฀presents฀advances฀in฀a฀conservation฀and฀reintroduction฀program฀of฀Sophora฀toromiro฀(Phil.)฀Skottsb.฀A฀ status฀quo฀of฀specimens฀growing฀in฀Chile฀is฀compiled฀and฀propagation฀of฀germplasm฀is฀performed฀by฀inter-specific฀ grafts฀ (scion฀ and฀ rootstock฀ are฀ different฀ species).฀To฀ date,฀ over฀ 300฀ grafts฀ are฀ ready฀ for฀ the฀ establishment฀ of฀ a฀ multipurpose฀Clonal฀Seed฀Orchard฀(CSO)฀with฀a฀germplasm฀collection฀based฀on฀three฀lines฀known฀as฀the฀National฀ Botanic฀Garden฀of฀Viña฀del฀Mar฀(JBV),฀Goteborg฀(Got)฀and฀Titze฀(Tit).฀Additionally,฀the฀first฀controlled฀pollination฀ seeds฀were฀obtained฀in฀order฀to฀understand฀basic฀aspects฀of฀reproductive฀biology.฀Activities฀and฀a฀work฀program฀ to฀ensure฀gradual฀recovery฀of฀S.฀toromiro฀in฀Rapa฀Nui฀are฀identified.฀This฀is฀the฀second฀documented฀experience฀of฀ rescuing฀a฀species฀listed฀as฀extinct฀in฀the฀wild฀by฀a฀traditional฀but฀effective฀technique฀in฀the฀field฀of฀horticulture,฀the฀ first฀being฀made฀with฀Molokai฀koki‘o฀(Kokia฀cookei฀O.฀Deg). En฀este฀documento฀se฀entregan฀avances฀de฀un฀programa฀de฀conservación฀฀y฀reintroducción฀de฀Sophora฀toromiro฀ (Phil.)฀Skottsb.฀Se฀registra฀el฀estado฀de฀ejemplares฀creciendo฀en฀Chile฀y฀se฀realiza฀una฀masificación฀de฀germoplasma฀ mediante฀injertos฀inter฀específicos฀(injerto฀y฀patrón฀son฀especies฀diferentes).฀A฀la฀fecha฀se฀cuentan฀con฀más฀de฀300฀ ejemplares฀para฀el฀establecimiento฀de฀un฀huerto฀semillero฀clonal฀(multipropósito),฀con฀una฀colección฀de฀germoplasma฀ basado฀en฀฀tres฀líneas฀conocidas฀que฀son:฀el฀Jardín฀Botánico฀Nacional฀de฀Viña฀del฀Mar฀(JBV);฀Goteborg฀(Got)฀y฀ Titze฀(Tit).฀Adicionalmente,฀se฀obtuvieron฀las฀primeras฀semillas฀de฀polinizaciones฀controladas,฀con฀el฀fin฀de฀entender฀ los฀aspectos฀básicos฀de฀su฀biología฀reproductiva.฀Se฀identifican฀actividades฀y฀programas฀de฀trabajos฀para฀asegurar฀ la฀recuperación฀gradual฀de฀S.฀toromiro฀en฀Rapa฀Nui.฀Esta฀es฀la฀segunda฀experiencia฀documentada฀de฀rescate฀de฀una฀ especie฀catalogada฀como฀extinta฀en฀su฀hábitat฀natural,฀mediante฀una฀técnica฀no฀tradicional฀pero฀eficaz฀en฀el฀ámbito฀ de฀la฀horticultura,฀siendo฀la฀primera฀la฀que฀fue฀llevada฀a฀cabo฀por฀Molokai฀koki‘o฀(Kokia฀cookei฀O.฀Deg). Introduction tree฀ that฀ is฀ distinctive฀ for฀ its฀ high฀ wood฀ density฀ and฀ was฀used฀by฀people฀as฀an฀energy฀source฀as฀well฀as฀raw฀ material฀for฀the฀manufacture฀of฀ritual฀artifacts฀(Oyarzun,฀ 1921;฀Palmer฀1870;฀Pinart฀1877)฀and฀the฀tablets฀known฀ as฀kohau฀motu฀mo฀rongorongo฀(Englert฀1988). The฀ last฀ native฀ specimen฀ of฀ this฀ species฀ grew฀ on฀ the฀slopes฀of฀Rano฀Kao฀and฀was฀cut฀down฀after฀a฀seed฀ collection฀made฀by฀Heyerdahl฀in฀1956.฀Today,฀a฀small฀ population฀of฀re-introduced฀material฀is฀cultivated฀on฀the฀ island฀by฀Sonia฀Haoa฀(Mata฀Ki฀te฀Rangi฀Foundation).฀ This฀ paper฀ presents฀ a฀ new฀ procedure฀ to฀ increase฀ the฀ success฀of฀reintroduction฀฀of฀this฀species. Flora฀growing฀on฀Oceanic฀islands฀is฀of฀particular฀interest฀ because฀of฀their฀endemic฀status฀and฀evolutionary฀aspects฀ of฀the฀genera฀present฀on฀these฀islands.฀These฀features฀ have฀ led฀ to฀ the฀ designation฀ of฀ a฀ hotspot฀ in฀ the฀ South฀ Pacific฀by฀Myers฀et฀al.฀(2000).฀Easter฀Island,฀or฀Rapa฀ Nui,฀is฀an฀island฀belonging฀to฀the฀Chilean฀State฀since฀ 1888,฀and฀is฀located฀about฀3,765km฀west฀of฀continental฀ Chile.฀The฀ fragility฀ (insularity)฀ of฀ its฀ ecosystems฀ and฀ the฀ human฀ intervention฀ experienced฀ throughout฀ the฀ past฀ 500฀ years฀ gives฀ us฀ a฀ meadow-like฀ landscape,฀ dominated฀by฀grasses฀and฀sedges.฀The฀flora฀of฀Rapa฀Nui฀ is฀made฀up฀of฀49%฀of฀introduced฀species,฀whose฀origin฀ dates฀ from฀ the฀ initial฀ colonization฀ by฀ ethnic฀ Rapanui฀ and฀ also฀ increased฀ in฀ the฀ mid-nineteenth฀ century฀ to฀ the฀present฀(Ibáñez฀et฀al.฀2001;฀Moreira-Muñoz฀2007;฀ Zizka฀1991).฀The฀same฀authors฀note฀that฀only฀7.7%฀of฀ the฀species฀are฀endemic.฀ The฀flora฀of฀Rapa฀Nui฀has฀lost฀25฀species.฀Among฀ these฀is฀the฀Sophora฀toromiro฀(Phil.)฀Skottsb.,฀a฀small฀ Background I)฀฀Ex฀situ฀germplasm฀in฀Chile.฀Maunder฀et฀al.฀(1999)฀ reports฀five฀different฀lines฀of฀฀S.฀toromiro฀growing฀in฀ Chile฀ by฀ the฀ time฀ of฀ the฀ study.฀ Currently,฀ only฀ three฀ lines฀can฀be฀found:฀The฀National฀Botanical฀Garden฀of฀ Viña฀del฀Mar฀(JBV),฀Titze฀(Tit),฀and฀Goteborg฀(Got).฀ Only฀ the฀ first฀ line฀ is฀ documented฀ with฀ regard฀ to฀ the฀ Jaime Espejo | Programa de Postgrado. Facultad de Ciencias Forestales, Universidad de Concepción, Victoria 631, Casilla 160-C. Correo 3. Concepción, Chile Francisco Rodriguez | Gerencia Tecnología Silvícola. Forestal Mininco S.A. Avenida Alemania # 751. Casilla 399. Los Angeles, Chile Rapa฀Nui฀Journal 5 Vol.฀27฀(2)฀October฀2013 New฀insights฀in฀conservation฀of฀Sophora฀toromiro฀(Phil.)฀Skottsb. year฀ of฀ harvesting฀ and฀ the฀ identity฀ of฀ the฀ collector.฀ The฀ same฀ authors฀ determined฀ by฀ molecular฀ analysis฀ (RAPD,฀ ISSR)฀ a฀ greater฀ genetic฀ variability฀ for฀ these฀ copies฀when฀compared฀with฀plants฀growing฀in฀Europe฀ and฀Australia฀(Maunder฀et฀al.฀2000). The฀ soils฀ of฀ the฀ island฀ are฀ a฀ volcanic฀ series฀ and฀ at฀ least฀ thirteen฀ soils฀ of฀ Rapa฀ Nui฀ are฀ cataloged฀ according฀to฀studies฀by฀the฀Development฀Corporation฀ (CORFO).฀Most฀have฀a฀process฀of฀“base฀wash”฀given฀ the฀percolation฀which฀is฀favored฀by฀the฀recorded฀annual฀ rainfall฀on฀Rapa฀Nui฀(Rivera฀2003). II)฀Experience฀using฀grafts฀for฀conservation฀purposes.฀ There฀ are฀ few฀ experiments฀ reported฀ in฀ the฀ literature฀ on฀the฀use฀of฀grafting฀as฀a฀means฀of฀conservation฀for฀ extinct฀plants฀in฀their฀natural฀habitat,฀with฀the฀exception฀ of฀ Kokia฀ cookei฀ O.฀ Deg.฀This฀ tree฀ became฀ extinct฀ by฀ human฀ action฀ in฀ Moloka‘i฀ (Hawai‘i),฀ and฀ has฀ been฀ recovered฀ using฀ Kokia฀ drynarioides฀ (Seem)฀ Lewt.฀ as฀ rootstock.฀The฀ limitation฀ to฀ using฀ grafts฀ is฀ that฀ these฀ do฀not฀show฀blooming฀events฀(Mehroff฀1996).฀Specific฀ inter฀grafts฀appear฀as฀an฀effective฀horticultural฀tool฀and฀ their฀use฀is฀only฀justified฀for฀species฀which฀have฀gone฀ extinct฀in฀the฀wild฀(Guerrant฀1996). Establishment฀and฀Maintenance฀Proposals The฀preservation฀of฀specimens฀in฀Chile฀is฀guaranteed฀ by฀the฀Botanical฀Garden฀of฀Viña฀del฀Mar฀and฀private฀ collections.฀ However,฀ the฀ big฀ challenge฀ is฀ still฀ the฀ reintroduction฀of฀S.฀toromiro฀to฀Rapa฀Nui.฀In฀1994,฀the฀ Toromiro฀Management฀Group฀(TMG)฀initiated฀actions฀ and฀ main฀ lines฀ of฀ research฀ for฀ the฀ reintroduction฀ of฀ this฀species,฀however,฀they฀did฀not฀consider฀a฀grafting฀ mode฀for฀both฀the฀formation฀of฀a฀breeding฀arboretum฀ as฀germplasm฀or฀CSO฀with฀seed฀production฀objectives฀ at฀the฀operational฀level. Based฀on฀these฀proposals฀and฀progress฀in฀various฀ fields฀ of฀ forestry,฀ Forestal฀ Mininco฀ S.A.฀ (CMPC฀ holding)฀revised฀the฀proposals฀of฀TMG฀and฀is฀currently฀ developing฀ activities฀ and฀ actions฀ (Table฀ 1)฀ to฀ ensure฀ the฀ establishment฀ of฀ S.toromiro฀ on฀ Rapa฀ Nui฀ in฀ the฀ medium฀term. III)฀฀Clonal฀Seed฀Orchard฀(CSO).฀The฀direct฀benefit฀ of฀forest฀orchards฀is฀the฀production฀of฀seeds,฀a฀product฀ of฀high฀economic฀value.฀The฀management฀and฀success฀ of฀the฀CSO฀lies฀in฀the฀knowledge฀of฀the฀reproductive฀ biology฀ of฀ the฀ species,฀ as฀ appropriate฀ horticultural฀ practices฀ play฀ a฀ key฀ role฀ in฀ productivity฀ (Sedgley฀ &฀ Griffin฀ 1989).฀ The฀ CSO฀ design฀ requirement฀ reflects฀ the฀ type฀ of฀ seed.฀ Usually,฀ when฀ producing฀ openpollinated฀ seed฀ available฀ as฀ grafts฀ or฀ ramets,฀ the฀ design฀is฀random฀and฀arranged฀in฀blocks.฀In฀the฀case฀ of฀ controlled฀ pollination,฀ the฀ designs฀ are฀ prepared฀ to฀ maximize฀ the฀ productivity฀ of฀ the฀ crosses.฀ Thus,฀ in฀ the฀last฀generation฀of฀forest฀seed฀orchards,฀grafts฀are฀ found฀ in฀ linear฀ arrangements฀ or฀ in฀ blocks฀ (Espejo฀ et฀ al.฀ 2001).฀ Regarding฀ the฀ choice฀ of฀ site,฀ the฀ orchard฀ should฀ preferably฀ be฀ located฀ close฀ to฀ a฀ water฀ source฀ in฀the฀case฀of฀intensive฀management฀(fertilization฀and฀ irrigation).฀Also,฀a฀good฀quality฀soil฀must฀be฀selected.฀ Soils฀ that฀ are฀ rich฀ in฀ organic฀ content฀ are฀ necessary฀ for฀the฀smooth฀development฀of฀the฀grafts฀and฀also฀to฀ ensure฀good฀seed฀production. Grafts For฀ rootstock฀ selection,฀ we฀ used฀ a฀ species฀ related฀ to฀ S. toromiro,฀ in฀ this฀ case฀ Sophora฀ cassioides฀ (Phil.)฀ Sparre.฀ The฀ species฀ has฀ a฀ wide฀ distribution฀ (easy฀ access฀ to฀ germplasm)฀ in฀ south-central฀ Chile฀ and฀ so฀ it฀ is฀ adapted฀ to฀ different฀ soil฀ conditions.฀ It฀ grows฀ between฀ 5-900m฀ above฀ sea฀ level,฀ but฀ its฀ habitats฀ are฀ wet฀ravines฀and฀deep฀soils฀near฀streams,฀rivers฀or฀lakes฀ that฀can฀be฀found฀in฀areas฀with฀rocky฀outcrops.฀There฀ are฀other฀species฀growing฀on฀Oceanic฀islands฀such฀as฀ Sophora฀ fernandeziana฀ (Phil.)฀ Skottsb.฀ and฀ Sophora฀ masafuerana฀ (Phil.)฀ Skottsb.,฀ however,฀ they฀ were฀ discarded฀ because฀ of฀ low฀ numbers฀ of฀ specimens฀ and฀ Table฀1.฀ Activities฀and฀actions฀of฀฀S.฀toromiro฀conservation. Activity Time (horizon) Advances Observation Propagation by grafting Short term Over 300 grafts. Mean set 70% survive Lines used Jardin Botánico Nacional Viña del Mar (JBV); Goteborg (Got) and Titze (Tit) Reproductive biology: (pollen management, stigma receptivity etc…) Medium term (3 years) 150 seeds by self pollination (68% of germination) Pollination in grafts of JBV and Got Establishment Clonal Seed Orchard/arboretum Medium term Ongoing (2012) Two sites: Easter Island and a continental back up Orthopedics* and induction (Paclobutrazol) Medium term (3 – 5 years) *crown management Rapa฀Nui฀Journal 6 Vol.฀27฀(2)฀October฀2013 Jaime฀Espejo฀and฀Francisco฀Rodriguez Table฀2.฀ Summary฀of฀grafts฀by฀year. Figure฀1.฀ Number฀of฀seeds฀and฀germination฀produced฀by฀control฀crosses฀in฀the฀2010/2011฀season. Figure฀2.฀ (a)฀Close-up฀of฀the฀graft฀area฀in฀the฀stem฀of฀S.฀toromiro.฀(b)฀and฀(c)฀Development฀stages฀of฀mini฀grafts.฀ Rapa฀Nui฀Journal 7 Vol.฀27฀(2)฀October฀2013 New฀insights฀in฀conservation฀of฀Sophora฀toromiro฀(Phil.)฀Skottsb. their฀ conservation฀ status฀ is฀ listed฀ as฀ endangered฀ by฀ Chilean฀environmental฀authorities. A฀ great฀ advantage฀ of฀ choosing฀ the฀ species฀ S. cassioides฀is฀associated฀with฀micro-organisms฀such฀ as฀bacteria฀of฀the฀genus฀Rhizobium฀and฀Mycorrhizae฀ (Zúñiga฀1996).฀This฀is฀of฀paramount฀importance฀for฀the฀ establishment฀and฀survival฀of฀the฀grafts฀in฀the฀current฀ conditions฀of฀the฀soil฀of฀Easter฀Island. The฀results฀to฀date฀deliver฀a฀71%฀survival฀or฀set฀for฀ the฀ cleft฀ graft฀ technique฀ (Table฀ 2).฀This฀ type฀ of฀ graft฀ calls฀for฀the฀insertion฀of฀a฀scion฀(semi-lignified)฀with฀a฀ similar฀diameter฀to฀the฀rootstock,฀previously฀cut฀at฀its฀ end฀(in฀the฀form฀of฀a฀bevel),฀and฀finally,฀the฀sealing฀of฀ the฀junction฀scion-holder฀with฀graft฀Parafilm™.฀Actions฀ for฀ex-situ฀germplasm฀collections฀or฀other฀lines,฀such฀ as฀Melbourne,฀are฀considered฀to฀increase฀the฀variability฀ of฀the฀future฀composition฀of฀the฀CSO. (photoperiod:฀16/8H;฀2.000฀Lux)฀in฀the฀laboratory.฀The฀ average฀germination฀obtained฀was฀of฀68%. Establishment In฀ addition฀ to฀ establishing฀ grafts฀ on฀ Rapa฀ Nui,฀ a฀ second฀ back฀ up฀ orchard฀ should฀ be฀ made฀ ฀in฀ benign฀ environmental฀ conditions฀ in฀ continental฀ Chile.฀ All฀ grafts฀must฀be฀established฀in฀boxes฀with฀the฀support฀of฀ gel,฀fertilizers฀and฀irrigation.฀The฀selected฀sector฀must฀ be฀ free฀ of฀ weed฀ competition฀ and฀ ideally฀ in฀ a฀ windprotected฀area.฀This฀can฀be฀provided฀by฀planting฀next฀ to฀established฀plantations฀of฀other฀species฀or฀by฀using฀ topographic฀depressions฀such฀as฀an฀area฀between฀two฀ hills.฀Alternatively,฀we฀may฀consider฀the฀establishment฀ of฀ manavai฀ (traditional฀ stone฀ vegetation฀ shelters)฀ within฀the฀canopy฀to฀leverage฀common฀plants฀in฀these฀ structures฀ as฀ in฀ the฀ case฀ of฀ Musa฀ spp.฀ in฀ the฀ town฀ of฀ Hanga฀Roa฀on฀Rapa฀Nui.฀Previous฀techniques฀known฀ as฀Inter-Situ฀(Maunder฀et฀al.฀2004)฀can฀deliver฀higher฀ spot฀ information฀ for฀ S.฀ toromiro฀ development฀ and฀ indications฀of฀future฀pilot฀plantations. Flowering฀Pattern฀and฀Seed฀Germination The฀ flowering฀ of฀ this฀ species,฀ as฀ recorded฀ inside฀ greenhouses,฀ starts฀ in฀ mid-August฀ and฀ finishes฀ in฀ November.฀ The฀ post฀ grafting฀ earliest฀ flowering฀ has฀ already฀occurred฀in฀the฀Titze฀line฀and฀was฀observed฀for฀ three฀years฀for฀the฀JBV฀and฀the฀Got฀lines.฀S.฀toromiro฀ blooming฀has฀a฀similar฀pattern฀found฀in฀the฀phenology฀ of฀the฀other฀Sophora฀spp.฀growing฀in฀Chile,฀in฀which฀ the฀peak฀occurs฀in฀the฀Southern฀Hemisphere฀spring. In฀ relation฀ to฀ reproduction฀ strategy฀ mechanisms,฀ apparently฀ S.฀ toromiro฀ presents฀ protandry;฀ however,฀ studies฀ are฀ required฀ to฀ clarify฀ the฀ foregoing,฀ because฀ it฀is฀an฀imperceptible฀change฀in฀the฀morphology฀of฀the฀ stigma฀ that฀ is฀ also฀ quite฀ small฀ and฀ undifferentiated.฀ To฀ this,฀ we฀ can฀ add฀ that฀ exudates฀ are฀ not฀ detected.฀ Studies฀ concerning฀ the฀ timing฀ of฀ receptivity฀ verify฀ that฀with฀support฀agents,฀stigmatic฀receptivity฀markers฀ will฀be฀made฀for฀present฀and฀future฀seasons฀according฀ to฀ protocols฀ proposed฀ by฀ Dafni฀ (1992)฀ and฀ Kearns฀ &฀ Inouye฀ (1993).฀ Parallel฀ management฀ studies฀ are฀ considered:฀ pollen฀ collection,฀ germination฀ tests,฀ viability,฀and฀storage. In฀ the฀ 2010฀ season,฀ self-pollinations฀ were฀ performed฀using฀anthers฀(with฀pollen฀dehiscence฀presence)฀ which฀were฀removed฀with฀the฀aid฀of฀fine฀forceps฀and฀ placed฀on฀the฀stigma.฀This฀work฀was฀performed฀every฀ three฀ days฀ during฀ the฀ flowering฀ period฀ of฀ the฀ grafts.฀ The฀use฀of฀this฀methodology฀allowed฀the฀development฀ of฀pods฀and฀seeds,฀as฀indicated฀in฀Figure฀1. The฀seeds฀obtained฀were฀soaked฀with฀sulfuric฀acid฀ (95%)฀for฀20฀minutes฀according฀to฀the฀protocol฀reported฀ for฀this฀species฀(Gonzalez฀et฀al.฀2008).฀Then฀the฀seeds฀ were฀washed฀in฀water฀and฀placed฀in฀Petri฀dishes฀lined฀ with฀ moistened฀ paper฀ for฀ a฀ period฀ of฀ two฀ weeks฀ in฀ conditions฀of฀stable฀temperatures฀(25ºC฀±฀2)฀and฀light฀ Rapa฀Nui฀Journal Orthopedics฀and฀Induction Once฀ the฀ material฀ is฀ established฀ and฀ showing฀ good฀ adaptability฀ to฀ environmental฀ conditions฀ (in฀ the฀ 2nd฀ to฀ 3rd฀ year),฀ ramets฀ should฀ undergo฀ orthopedic฀ work฀ in฀ the฀ crown฀ in฀ order฀ to฀ obtain฀ a฀ greater฀ number฀ of฀ points฀ to฀ generate฀ multiple฀ buds฀ to฀ be฀ induced฀ with฀ Paclobutrazol.฀This฀ chemical฀ known฀ as฀ PP333฀ (2RS,฀ 3RS)฀-฀(4-chlorophenyl)฀-4,4-dimethyl-2-(1,2,4-triazol1-yl)฀pentan-3OL)฀has฀shown฀efficacy฀in฀a฀large฀number฀ of฀ fruit฀ and฀ forest฀ genera,฀ especially฀ Eucalyptus,฀ and฀ should฀ be฀ tested฀ (Sedgley฀ &฀ Griffin฀ 1989).฀ There฀ are฀ different฀ techniques฀ of฀ application฀ and฀ the฀ most฀ effective฀is฀soil-applied฀and฀is฀introduced฀to฀the฀plant฀ via฀the฀root฀system. Conclusions S.฀toromiro฀conservation฀does฀not฀rescue฀the฀ecosystem,฀ biodiversity,฀or฀lessen฀complex฀interactions฀with฀other฀ species,฀ per฀ se.฀This฀ effort฀ is฀ only฀ an฀ approximation฀ in฀ the฀ rescue฀ of฀ a฀ portion฀ of฀ collective฀ memory฀ for฀ Rapa฀Nui. Seed฀ production฀ by฀ maintaining฀ a฀ CSO฀ and฀ subsequent฀establishment฀must฀use฀integrated฀strategies฀ and฀ techniques฀ used฀ in฀ both฀ horticulture฀ and฀ forestry฀ along฀ with฀ some฀ ancestral฀ practices฀ proven฀ by฀ the฀ ancient฀ settlers฀ of฀ the฀ island.฀This฀ proposal฀ will฀ only฀ succeed฀ with฀ the฀ proactive฀ interaction฀ of฀ different฀ parties฀ such฀ as฀ private฀ companies,฀ government฀ agencies,฀NGOs฀and฀the฀community฀of฀Rapa฀Nui. 8 Vol.฀27฀(2)฀October฀2013 Jaime฀Espejo฀and฀Francisco฀Rodriguez The฀foregoing฀appears฀as฀valid฀and฀necessary฀for฀ restoration฀of฀this฀species.฀This฀is฀still฀in฀the฀development฀ stages฀and฀should฀be฀monitored฀for฀early฀feedback฀and฀ to฀validate฀the฀initial฀proposals.฀In฀our฀view,฀Rapa฀Nui฀is฀ presented฀as฀an฀opportunity฀that฀can฀be฀extrapolated฀for฀ particular฀cases฀of฀species฀that฀are฀in฀survival-limiting฀ conditions฀both฀in฀the฀wild฀and฀ex-situ. Maunder,฀M.,฀A.฀Culham,฀B.฀Alden,฀G.฀Zizka,฀C.฀Orliac,฀W.฀ Lobin,฀ A.฀ Bordeau,฀ J.M.฀ Ramirez฀ ฀ &฀ ฀ S.฀ GlissmannGough.฀2000.฀Conservation฀of฀the฀Toromiro฀Tree:฀Case฀ Study฀in฀the฀Management฀of฀a฀Plant฀Extinct฀in฀the฀Wild.฀ Conservation฀14(5):1341-1350. Maunder,฀M.,฀E.O.฀Guerrant,฀K.฀Havens฀&฀K.W.฀Dixon.฀2004.฀ Realizing฀ the฀ Full฀ Potential฀ of฀ Ex฀ Situ฀ Contributions฀ to฀ Global฀ Plant฀ Conservation.฀ In฀ Ex฀ Situ฀ Plant฀ Conservation:฀Supporting฀Species฀Survival฀in฀the฀Wild.฀ E.O.฀Guerrant,฀K.฀Haens฀&฀M.฀Maunder฀(eds.):389-418.฀ Washington,฀D.C.:฀Island฀Press.฀ Mehrhoff,฀ L.A.฀ 1996.฀ Reintroducing฀ Endangered฀ Hawaiian฀ Plants.฀ In฀ Restoring฀ Diversity:฀ Strategies฀ for฀ Reintroduction฀of฀Endangered฀Plants.฀D.฀Falk.,฀C.฀Millar฀&฀M.฀ Olwell฀(eds.):101-120.฀Washington,฀D.C.:฀Island฀Press.฀ Myers,฀ N.,฀ A.R.฀ Mittermayer.฀ C.G.฀ Mittermayer,฀ G.A.B.฀ da฀ Fonseca฀ &฀ J.฀ Kent.฀ 2000.฀ Biodiversity฀ hotspots฀ for฀ conservation฀priorities.฀Nature฀403:฀853-858. Moreira-Muñoz,฀ A.฀ 2007.฀ Plant฀ Geography฀ of฀ Chile.฀ An฀ Essay฀ of฀ Postmodern฀ Biogeography.฀ Unpublished฀ PhD฀ Thesis.฀University฀of฀฀Erlangen-Nuremberg.฀ Oyarzun,฀A.฀ 1921.฀Toromiros฀ de฀ la฀ Isla฀ de฀ Pascua.฀ Revista฀ Chilena฀de฀Historia฀Natural฀XXV:133-140. Palmer,฀J.฀L.฀1870.฀A฀Visit฀to฀Easter฀Island,฀or฀Rapa฀Nui,฀in฀ 1868.฀Proceedings฀of฀the฀Royal฀Geographical฀Society฀of฀ London฀14(2):108-120.฀ Pinart,฀ A.฀ 1877.฀ Voyage฀ to฀ Easter฀ Island฀ –฀ 1877.฀ In฀ Early฀ Visitors฀ to฀ Easter฀ Island฀ 1864-1877.฀ The฀ Reports฀ of฀ Eugene฀ Eyraud,฀ ฀ Hippolyte฀ Roussel,฀ Pierre฀ Loti฀ and฀ Alphonse฀ Pinart.฀ G.฀ Lee,฀ A.M.฀ Altman฀ &฀ F.฀ Morin฀ (eds.):106-136.฀Los฀Osos:฀Easter฀Island฀Foundation.฀ Rivera,฀Fernandez,฀E.฀2003.฀El฀Componente฀Forestal฀en฀los฀ Sistemas฀ de฀ Producción฀ de฀ los฀ Pequeños฀ Propietarios฀ de฀ la฀ Etnia฀ Rapa฀ Nui฀ de฀ Isla฀ de฀ Pascua,฀ V฀ Región.฀ Unpublished฀ thesis.฀ Facultad฀ de฀ Ciencias฀ Forestales,฀ Departamento฀ de฀ Manejo฀ de฀ Recursos฀ Forestales.฀ Universidad฀de฀Chile.฀ Sedgley,฀M.฀&฀R.Griffin.฀1989.฀Sexual฀Reproduction฀of฀Tree฀ Crops.฀Waltham:฀Academic฀Press.฀ Toromiro฀ Management฀ Group.฀ 1998.฀ Minutes฀ of฀ 5th฀ International฀ Toromiro฀ Management฀ Group฀ Meeting.฀ Available฀ at฀ http://www.kew.org/conservation/cpdu/ Toromiro/tor_min1.htm฀ Zizka,฀ G.฀ 1991.฀ Flowering฀ Plants฀ of฀ Easter฀ Island.฀ In฀ Palmarum฀ Hortus฀ Francofurtensis,฀ Scientific฀ Reports฀ 3:1-108.฀Frankfurt฀am฀Main:฀Palmengarten. Zúñiga,฀ E.฀ 1996.฀ Asociaciones฀ simbióticas฀ tripartitas฀ en฀ Sophora.฀Unpublished฀thesis.฀Conducente฀a฀Licenciatura฀ en฀Biología,฀Universidad฀Austral฀de฀Chile.฀ Acknowledgements The฀ authors฀ acknowledge฀ the฀ suggestions฀ and฀ corrections฀of฀Ana฀Betty฀Haoa฀Rapahango฀and฀Sonia฀ Haoa฀ Cardinali฀ and฀ two฀ anonymous฀ reviewers.฀ This฀ document฀corresponds฀to฀the฀oral฀presentation฀made฀in฀ “Evolution฀of฀Life฀on฀Pacific฀Islands฀and฀Reefs:฀Past฀ Present฀ and฀ Future”฀ presented฀ in฀ Honolulu,฀ Hawai‘i,฀ 2011.฀Additionally,฀J.฀Espejo฀wishes฀to฀thank฀Forestal฀฀ Mininco฀for฀its฀support฀฀and฀฀the฀execution฀of฀all฀actions฀ of฀the฀฀S.฀toromiro฀฀rescue฀program. References Dafni,฀A.฀1992.฀Pollination฀Ecology.฀A฀Practical฀Approach.฀ Oxford:฀Oxford฀University฀Press.฀ Englert,฀ S.฀ 1988.฀ La฀ Tierra฀ de฀ Hotu฀ Matu‘a.฀ 4th฀ Edition.฀ Santiago:฀Editorial฀Universitaria. Espejo,฀ J.,฀ J.฀ Harbard,฀ &฀ R.฀ Griffin.฀ 2001.฀ Consideraciones฀ operacionales฀ de฀ cruzamientos฀ controlados฀ masales฀ (OSP)฀ de฀ Eucalyptus฀ globulus฀ (Labill)฀ en฀ cuatro฀ temporadas฀ en฀ Forestal฀ y฀Agrícola฀ Monte฀Aguila฀ S.A.฀ Valdivia:฀Actas฀ Desarrollando฀ el฀ Eucalipto฀ del฀ Futuro.฀ Simposio฀Internacional,฀IUFRO. Guerrant,฀E.O.฀1996.฀Designing฀Populations:฀Demographic,฀ Genetics,฀ and฀ Horticultural฀ Dimensions.฀ In฀ Restoring฀ Diversity:฀Strategies฀for฀Reintroduction฀of฀Endangered฀ Plants.฀ D.A.฀ Falk.,฀ C.฀ Millar฀ &฀ M.฀ Olwell฀ (eds.):171207.฀Washington,฀D.C.:฀Island฀Press.฀ Gonzalez,฀ M.,฀ I.฀ Quiroz,฀ E.฀ Garcia฀ &฀ B.฀ Guitierrez.฀ 2008.฀ Escarificacion฀ química฀ con฀ acido฀ sulfúrico฀ como฀ tratamiento฀ pregerminativo฀ para฀ semillas฀ de฀ Toromiro฀ (Sophora฀ toromiro฀ Skottsb.)฀ Ciencia฀ e฀ Investigacion฀ Forestal฀14(1):111-118. Ibañez,฀P.,฀A.฀Bordeau฀&฀M.฀Rauch.฀2001.฀Árboles,฀฀Recursos฀ Naturales฀y฀Comunidades฀Indígenas฀en฀Chile.฀Lecturas.฀ Corporación฀Forestal฀Nacional.฀36-44. Kearns,฀C.A.฀&฀D.W.฀Inouye.฀1993.฀Techniques฀for฀Pollination฀ Biologists.฀Boulder:฀University฀Press฀of฀Colorado.฀ Maunder,฀ M.,฀ A.฀ Culham,฀ A.฀ Bordeau,฀ A.฀ Guillames฀ &฀ M.Wilkinson.฀1999.฀Genetic฀Diversity฀and฀Pedigree฀for฀ Sophora฀Toromiro฀(Leguminosae):฀A฀Tree฀Extinct฀in฀the฀ Wild.฀Molecular฀Biology฀8:725-738. Rapa฀Nui฀Journal This฀article฀has฀been฀peer-reviewed.฀ Received฀1฀July฀2013;฀accepted฀25฀August฀2013. 9 Vol.฀27฀(2)฀October฀2013