5 Developments in Crop Science 12 Applied Mutation Breeding for Vegetatively Propagated Crops C.BroertjesChaper « SHOOT APICES: ORGANIZATION AND POST-IRRADIATION BEHAVIOUR For many years the postion ofa mutate celina plat hasbeen considered as of late importance forthe ure ate ofthat al. Often a itneon wis ade ‘eee the postion of such els in meristems and in ferent isso, Mute sions in ells outside messes ware si to be without pret aly sce, Bg surrounded by many unchanged cls, they canna be deesed and maitaned The fame vew predominate concerning muraion in dividing merce outide the cot apex "eat about the nontance ofthe psilon ofa mutated ctl have changed rather rasta throughout the ast decade, Fst ofl are has bon te inoue tion ofa practical approach to mutation brescing: the so-called sdrettous ba ‘echnige” By this technique elle, ofenofepdetial ori, canbe sutlael 10 eterna, and o Geveop ino a mersems and, subeequeny, ino an apical bad, This avenidous bud chau, invivo a inv, willbe futher cused in Chapters. ‘Gradually twas discovered that other deenitd cls could aio be stilted to develop msi act and, with he ncened interest te ave of i is ton methods, i was found han a ever neresing our of pant spies (or tases can be regeoeated,alvgh reesneraioneapacyiney vary Wide. Despite these developments, there stl ase net i he beaviou of aot apices. Thuy offer the (mutation) trader an sary ad sale system for proseediog ‘fom a single (spontaneous or induced) mata cll in aon mated ase towards * structure witha much ger mated are. Aplcesarying both mulated an som Iutated elo» srer may Belpo laces the sight of developmental Botan Ita shoot apex tehavour ‘In the following scions the thor! backgrounds of shoot apex behaviour, shimerism and reated subject are dssased. No al scons are equally televanh deseibed bere wil help to preven mistakes in planing Inuaion breeding pro This secon is meant to provide basic information about the organization of| BF soot apices, as far as thsi of eevace to nation breeding. To heft eon [+ of oar Book we referred tare ate by Gilford sed Coton (1971) a5 food [2° Sarng point for further reaing on this topes Tis adc bs vad, dee ‘merous aditonal publications that have een ited since hen. This does notx» infer that no new views have been formated aboat shoot apices. The baie cm. jes of some decades ago, homer have no bien abandoned ye, end discussions Shoot confcing views oF ttrprtatons sll go on, From te pint of view o Predict aod explain experimental finns Sa ace thesractre of ep! bods, airy bos fe ined unless squad otteribe Advetitous Bute can als be uded, but in that ease it must Secures hari tates some time before one or fe’ originating cals cn ie se To nable apical stn with diffrent layers end pater of zonation 8 oust Different words a often sie fo inde the apical area of «shoot. Dees (030, for ecampley sntions eons pla, shoot shoot pes epic meristem ntact dome, os ten the ferme thoot apex and pial bud ae Wed in 8 air penevel sess idcate the ae ofthe ste that is stl to Ue youngest nda aes Getelop, but thay once orga these meristems ae very stable *Ciang and animated disso ssl going om eoacering "ial so a0 pox, Sty speaing a ial el i cll which divides into wo dager cel he of which remain in the fixe apical positon, the other Being added 10 he ‘Srncmatie sue Esa, 1965), The question of whether such ital es ally ‘ew iso det importance fo mstion war, Because induction of a mutation i Sh Sai eel means tat & permanent lbeage of mutated dase el il te produced by that eal In he cage where occupation ofthe apical positon not euannt is matted ial cll shift the Danks witout being relaced Ey one a he mutated daughter cel, another aon-atated inal cll wl ARE XS Poa tad the prodston ofa ineage of rated cls wl be ternated, unos re ahould be permanent inal elm the ans ee below). is 1853, Hormecr postulated fhe existence of Sige nil clin phanero ams Atcrthatr many investiguors ted fo provide microscopical pro0! of hs bt thy failed oo soa ost cel the ape repo lok very ier. However Fiofmelaers concept hes received considerable support over the seers. 10 1868 Hanucin presented his Hatogen Theory According to his theory, some Kind of Sratfinfon ess in apes of angiosperms plats. A cena or (corpus of ‘oul avrenged colo was said to be surounded by a number of eglar, mae [ilayte Each yer and te compa are sudo be ded From a eis which US found veteally superinpoued on each oter, atthe uinate ip ofthe ail ‘Seep the aconly predestined oe ofthe ilfren growth ayers oF sone sexed to them by Hlastcin. Aceovdag 19 Newrnan (965), Hanstin’sollal wa pa forward by Buder (1928) and by Seid (1928) this “Twnia-Corpes Theory several tna layers ar ingushed, the layers being charcterzed by layer. Be didine Scheuat Vea Gut rate 28 sulle shown Planeta eotyled "The F ssihey ean,pot cals ‘nina si ivsons. The plane of dvvon cases cera independence ofeach layer. Below the tunica layers a cre, the earpy found whi cons cle ving in an ireeions. The Tunea.Corpae Faery docs 9 neces app, the tonics layers remain completely dscrte throughout te ves, Feil! dvs sions in tunica layers are meine observed ‘At present most authors stl comsier the basic eas of the Torca-Corpur ‘Tory tobe vali. Support for the geeral corres the opinions of Boge sod Schmiat Is mosty derived from histlosie! werk and from sedi on chinese otbl (sable) prising ploidy chimeras erochimrss) or paid cera Ove he years & numberof modifications have ben suggrte and same authors Cs; ‘Yon Guttenberg (1960, have completely rejected the theory bese Soest Satna «al. (1940 and Sana and Blake (1941) proposed the now seaclly io expressions Ll, Ei aad Lt inde the effet erm lays oleae cock ‘he nue a gem layers snd the numberof cll layers within a germ ayer ste subject of controversy. Dern (1951) ees to pris "Histogee age 2 “histones” which he bellews to Be present in perhaps all tnpaspermy. Ia i abs the lier appearance ofa sbo0t apex of potato (Solanum ubrosn 6 ‘Another concept of zonation was proposed by a French siool. According to Planefol (97), obar eles terminate in meristematic tsue sated at see sistance fom the apical tip. The author lied hat thee ee, whch sve called the “Yanacau ina”, aethe rea inal cel, No tuna were eto be peat ‘the summit of he axis during the vegetative phate of devlopnent teoiledonous plants. Buvat (1952) calle this sree “missin eatin” oe ‘rating mvistem Pig, 6 becuse, according fo French pinion, no divin <2Fin that reson during the vegeiatve phase, In 1955, Bower, Bure ao aden ‘ed that vegetative cell sotcsines show dvion. The French concept was not enely new Foser (1942) and Majumdar (1942), for example, had already demonstrated tat shot spies ae nots homoqeneoeg they were previously believed tobe Both athors refered to 4 etohisoloneal onion tat ean often be obeered ig. #). Even ear, Grégite (1938) reed "hat diferent stages of development may lea to iferences the sone pence ot“The French dea have ben cad om several occision, 4 by GiTord (1954, ‘ut sine Boat (1955 has edited that te orsnal concept was to ii, seems thatthe diferent views in fact ate reasonaly compa ‘ussow seerallyacepted that cls at the smi of vegetative apices 20 aso sive, but at lowe rate tan hoses the ani. Moh consoversy ales ‘out the roe, postion and namber of la Mos authors now are of te opinion that en” nal cls do no ex (Dui, 1990), The aie] mess fsld to be ade up of dynamic popation of els and to have selvorganizing properties Soma, 197, Ball, 19%). I cannot be Selo, however, that some kind of an ulate sores of piary growth mast exist “The most elabe method of tavextentng the pein of ths source els ‘at are ake in one way or another Flloine ths method Stewart and Besse (0970) for expe, stated the dimensions of (merical) chlorophyll spe ine Seretplet species and thus ectermised the number of nal els nthe shoot ‘pes of those spaces, For Ligusrum tse conclusion Wes tha there vo os the intl cls per ner, nial al wn found to vide only once pron ‘mately 12 days ths division giving rs 10 ewo new cls the os istl daughter cell should then be considered the ne inital cel Ths low dain sats supe {0 mak iil calle te ulate sure of growth Bal (1974 describes labeling experiment by using puncturing, appetion of "Spang. ‘rc ‘ent,leaf are since 6 clonal propagation of sleet mutans especialy cing the eal years, i slow, ictake aproximaly $ yeas before enough plane ar nalable ead rough ek ‘pines of the mutans has bon gained just commercalexploation af ns ‘Adrentions bulbs are obtained In some ater bulb cops trom scooping the ‘bulbs ronal way of vegetative propeaton of or ane, hyo Ts rethod may have a wider aplication, e Musca sce he derimental eects ‘offre can be prevented very ffs bythe ue of fungicides, such tenons (Gale) (Broetfes and Alters, 170) or, even Der, by radiied sl (Bouts ‘nd Lock, 1980). Artieally made Pub tale obey cating bute mt Dies, aio offer posites in propagation for eat, aplication of the one, sce or woseae method (Luyten, 1935, Taub, 198, Alera, 1970. Syscns 1979 to species ke Armarylis(Hippasrumy, act, Fs, Nerina tlio soe dat fod, Whetier adventitious bulletin such ses oiiat ro one ell to produce noa-chimeral mutants fer Inada snot Ged) Hesen, Brit Deas guy ovo Table. Preliminary resle wits Amol show tha aparenly seid mista are produced onsale piers of izaited babs (C. roses, unpubl dat), Bit flso certain percentage of chimeras mas observes Senta maton beng bud tenia for mutation breeding in esetavely propagated orn plants ‘nas been found in Stepocurps, sing the calvar Conant Ny, Traced or ‘olehcne rated hufeves produces sriknly high percentage of tens oF Dolyplid, respectively. Appreximatly 95% ofthe adreniou tats produced after radiation withthe optimum Xray dss of 30 Gy were volte etme ofhaleaves for 6h wih 0.19 colbicee, over 208 othe panels were non =tochimerl tetrapods, In toa, 857 mate were ovained, ive of which were ‘old commercial within 3 years ofthe sat ofthe experiment (Bron: 16), ‘Similar resus wee obtained with Achienes. Thoush fewer tutta Were po ose then in Streprocarpus, raion of aves ofthe ear Pal Arnold wich 530 Gy Xerys produced @ umber of sé mutts, Within sly more than 3 Years of irradiation, several mutans were being commercial expatel, saely ‘Srngtne” and “Early Arnokt, both of which flower approinatey 3 wecks cate than “Pasl Amol’, and other marie ee Chepte secon on (sneriaene) (Brees, 19720), ‘Brown (19718 reported 30 seid and two chimera nvtants among 204 plans us- tng simular cultivars andthe same Xray doe a that mentioned hee ‘Broce and Leting (197) obaied ealisiey sold mutants fe the radia tion of Kalanchot leaves wich X-rays. Acvertous panes prection ws el ‘ower than in Gespesacee. Moreover, Kelenchoe pans tele shorty rest "en to ower which makes prog ith the programme relaely slow. Conse. ucnly, Cakes some longer (ES yas) before the fe mutants canbe ut Hundeeds of slid mutans of Ipomoes boat (awet potato) bave been po oced by iradiation of detached eaves. Several Of them mere promiong ehrespect to various characters, such as dry mater and proen content ski and Fes ‘Sibir eaoten) of the toberowt root typeof plat, and yield (Love, 1988 Wilson, 1513) (seston om set potato, Chapter 6. 1 Seta teaver fe tills hybrid. Several ofthe muaots Were ‘reninacl within 4 yeers (Doornbos and Kerper, 973: Mike eta 1978) “Papromia can aso be propagted easly ffom detached faves (Bigot and a dt Peer ebeaal vargas hus pops, ee cm ests tenor nos) compl chlropst le elon) plane te forme. ia om oneal The oucoe sana epernen ith refore "Ino tera! poo! (© oouer wnpusied d). Howeve, Beran ZAP span GOED) neem avetows sbots from Peperomia os bay Se anne frm epost cl nd tha the fc tat omshont cee oy ona tom ove cal (We we he word get, on Peo ‘Sis of one (notated) (epidermal el). Whatever explanation igh, the fact chat fhe doa evs of ea menier of be ices for example, Endo Sturt Ovntogsu na Sil, realy prosse avenius Seite saver sven flmiag ev optmam Xray dae he murain eqn mice spss vy fw a a consoqunnes of th ntl crops) The Thott Geserceae (, Bowes and ALY. Afkema, 1970, unpolished da) ‘aioe hanes sd mca have been one fer radiation of bab sels medatly hr sang, o ter esion of tl ust Defrag SS ronlag motets or tapes (tan afer cone eaten! of Sr Stu Rens proper for fri ein Ove ease, ae 2A, Rousran, on oange Howerag an ofr. Tabsco(C Brand A Bich Tule spied ta reasentous bol techie sapped i a experimen ik Noone an eer ac deed, nn ater adaion of deaced eve By SHR USES y he sun gent acgrund te ck plant a ere comcqenty stom inbeedng esc (De Netansou tal, 1971) Adventtous buds originating from more than one cell tn many eases its mot obvious, or even not to be expected, that the pies of adveaious bos rn ont ob sl It ow that buds on ems ad 00 fen develop endonenows ftom more tan one ee, probably from a rected b= pevane S988 P1935) ‘mately bably ctehat or 3). ‘fou By i Ay nurnber In Chrsonthemum,advenios bids developing on sems of de-budded young pvcoal plant, were eter st perinal or penataly uform (Stewart 8nd Dermen, 19704). The authors concladed thas the shoot bad gine rom {to or more cls which may of may not have derived em fren apc! layers 1s Poinsettia Euphorbia pulchrima) much larger sectors 290 eveh cme (Gercina) entans were obtained by aéentious bud formation fer reoval of "he teil bud as wala al lateral bods onthe radiated plants (Love 1972), In Gerbera a mishod in which mature planet praned of al leaves eh are Part ofthe wers and oo fas Ween usd. After aati, these plats” wee Placed in swarm aud bua greenhouse, Wa 34 months, «rea numberof ‘dormant buds had devetoped into shot the number ranged fom 10 to 2, dope ‘ent amongst ocr ting, on the avr. The small shoots wee co of rome tnd planed (Brenan, 1990, ‘Lange etoed and spparety slid mutans of potato cn be obtained in varios way, Ferwerda (1968) inated single pecs end among the mitts obtane, 2 slaively hh percecage (DW) seemed foe soli. Doria eer tber fadiated dey after bares, deed ter ples ind very sol, prematirely hatvestedtuters (25 mm) respecely showed gradually deining chimes. ‘Chimarsm ean aso te lited by growing saveniiows sprouts onthe fos of ‘radiated tuber cs whieh, ns hasbeen mentioned by Howard (96), develop ftom aesrited numba of cls (edema, 1967, 1972-6). However, the best Wa) ‘nownto obtain sls (on--adaton (60-100 Gy; 6 Gy) that mutton fe. ‘Guency increased wit incesing dae and was higher nthe ain sot ban he Steals (Bade and Esan, 1977; But and Ragazain, 165; Bute a. 1965 Drama ay 1969), ‘An iresng cera was wed by Preav-Leoy (1859, 1970, 1744 1973) 10 Cahir Jacqueline which has 2 genetalyyelow Land genetic red L-ll and TCIM By colchicine easnet, vo types of eyo-himeras wee obtaned, nares 4, 28 and 24,40, 4a (Ll, Ll and Ll, respective). From the colour and nea of the wer a well these of the sel mace conetsons about the ‘crign of shoots on arated nats cold be drawn, The orange lower slow, fo Sten, eesuts from the Influence on the synthesis of pirment inthe episetal fells ofthe mesophyll wich Carries the ed ower gonype Bute oot pigmented set (Une fs not alway the ene: in ite sports for fsance, Lt ' WhieaPd iy to find the o-chimeras 23, ae, 4-4) and 42-4 ong peel eh ‘igus. In other word, Lell and Ll are apparently slvayssenealy Metal, both beng ener diploid (422) of tevapiall 4. ‘Dommereves and illo sls we rciton (say cers. They found vas dons inthe efoto! Co yeaye (4, 60 or 80 Gy) onthe degree of wneverng bseve, supposedly asthe resul of ferences in dose rte (Dommerzues and ‘Gilt, 1965), The same sikors ner developed vivo and evra edveatous bud technics wheraby they eoulé produce genetically homogeneous pans, staring from chimes In then vivo metod, plants were topped and he wounded surf rated with 19 Indah batyae aed to Torn cals fom which shoots cole ainately grow. ne vitro method, explants of the apex were wed 0 rode fata from which adventitious shoots developed. When awit sport was ued, ap proximately 709 of te sheoe in iva and 100% of the shoot za weve now Ehimeal mite lowering pants (23 of 9 plants td neo nine lat, especie ¥) (Dommergucs sn Gil, 1973). “The sanfeace in carnation, of adentious shoots hs since been asc by several authors suck as Broarjes (196) and Casters (1978) whist SIvy (979) seb methods to produce abd propogats cer sable perielnal chimeras or Inport work for commersal) metaion beeing was cried out by Sparaai and conorker (Sparnai 1978; Sparnai and Dersiak, 197, 1971; Sanaa {aly 1940, They uly radated anooteé eatin, te lower 2~3 em of which was protected hy ea to entre maximum Foo formation. The doe wd ‘Rien he variably of quanatvecharaces such a yeld was 25~ 50 OY X-rays, wheres a bigher dose wat sasessary for Mower colour mations nee tablem for mutton bestest toad compromise Between the ecu ad of an experiment satay speaking ands practical possbiss Pes nen peonement of chraces tat are not dec visible s rene Sa oe he amount of material to be observed rapidly erases by om aerate propagation. Therefore, tf the utnost importance thst re aes ails rece the amount stan early age were developed by Faanee vere grouy. If seketing for increases ve, for instance, probable sear ie pesslected oa the bass of clonal yield data Sparaas, 1978). sa ey coredue econ ofthe exerts i to sles ery as possible witout “toanag foo many snes, Care (193) demonstate that he frst ance eee tars of the Second Fo of etngs fom ranted plans Pr EEYGINs or ine mana, Concson: ie tore cost efecive to irate more ‘isto and take emer ctng rom each lant, He also postulates that ration Syren Sane coud be mov eu than ication of lager murber of tain sar conlsion was aen by Ss and Menu (18). ss ns tha etoinion ean only be a oea co ierestasahliy Sem ota ong casting cane ae sed Tn young, new car the ERS 2 tural varity nthe terial large enough, and ot sicily Saag ey gradation, bu tes clr the esa election pred in On vara anual are stint to mpeve several quantitative characters (Cast ee aiough tha maybe rein ger doesnot ake info account he Tayktanec or the owner of ven cava of producing the necessary im spe raets hmsel before bs collngue and competitors fad (or produce) thet realest). We are teafore ofthe pinion tata breeder shold eee poate to induce he peceenry variably and improvements 8 Vy pro Tce noncoer, should be catiog oo a large sale, Deraus ofthe rani pees aoa bu technique foe the prosein, aswel as forthe rapid tna Tics af avenious bud scnigues, ee in vivo by dcaptsion of pant son OP roto oat sd mats om 8 uege comercial le, as 06 yet ben reported, however Se eh lined nur of cosineria! mutants has been produced or epued, Suapie te fet that eration was one ofthe it objets of radiation eee tad, Some mee oblaned by wradlating unrooted cutis witha ih TWitigo Go) of crap to lndue Mower colour changes. To preven inition of Tee SGusgeapacys te Towes pe of the cutis as shielded by a Ie pte. ‘hee mutants weve obitined by EMS teament of sllry pols end sezcton ces pans srown from the axilry shoots (Henrich and Gwe, 1982), smog iy we ange of lower colours was obtained frm te evar Korineona mee etnaChrysanthemum, ages oF the Composiae(Aseracee) family oni senual flower production. It ane of the most importat cot fewer crops in The ‘Neel, for lstance, over 700 10" flowers were sol 1985). ‘Chssonthemurn morifoium & a specie wid an extremely Tong History a8 2 «alias plant it wa rowan China drag the ne of Confucius, approximately £00 B.C. Berween 72 and 180 A.D. a few culvre were fmported int span wher in lr year, the lower became sacred. A he end ofthe 16th ceaury and {asng the 1 entry numberof forms were introduced 10 France and Great ‘Beal, Nowadays many reser in Western Europe, the U.S.A. pan and other form, de, production, quay, darsernstane snd Yection fo environment ow (npaatre tolerance, ail by ros breeding but ineeasin by indulog muta ‘Sone a ‘Sports te results of spontaneous mutations, have been reported by various suthor (Anderson, 1935 Dovrck, 1951, 1956: ak, 1955). Some cultivars have {hen ie toa great umber of pore sucha ‘Sweetheart, "The Pavourte’ (ARO et; Bowen eal, 182) and indanaple’ (Yoder Bros, Ine. 1967). Washer (0958 reported that approximately 3088 of te evar rated 3 spots te Ses orp by sport, however, soften ever large han the cage occupied BY ‘he vial seeing cuts. ‘Most mation testing has ben sad out with the hexeplo Chryanchemum ‘morfollum Ram. (60 = 4), abo refered to as C. x hovtorum or C. indicum ‘Some atenton has be paid othe economically uch ss importa dip see. propagated anbual spel: (2n = 1, soeh a8 C.carinatm Se futher on in his Peso Raditon induced spots have been reported by many authors: Bowen eal 1969; Brorts 1966, b Cavs, 196,196; Cha, 1956, Crandell eal, 196¢; Das eal, 1974 Ful and Matsui, 1967; Gupta, i966; Gupta and Shukla, 197 Masimora etal 961; Nakajins snd Kaveara, 1957: Nybom, 96; Ruprecht 196t; Satory, 1973; Sheherbakon, 1565; Shocsan atid Seen, 195% Simon 1967 Van Hose, 192 Yamagucht and Takato, 1970; the more recent Iter i