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Inhibitory Effect of Naphthylphthalamic Acid(NPA) on Stem Growth Induced by Auxin inPrecooled Tulip Bulbs

Saniewski, MarianDepartment of Plant Physiology, Research Institute of Pomology and Floriculture

Okubo, HiroshiLaboratory of Horticultural Science, Facility of Agriculture, Kyushu University

大久保, 敬九州大学農学部農学科園芸学講座

http://hdl.handle.net/2324/24250

出版情報：九州大学大学院農学研究院紀要. 43 (1/2), pp.59-66, 1998-11. Kyushu Universityバージョン：権利関係：

J. r ac. Agr. , KYlIshu \lniv., 43 (1 , 2). ;)9-(j6 (l!!!JH)

Inhibitory Effect of Naphthylphthalamic Acid (NPA) on Stem Growth Induced by Auxin in Precooled Thlip Bulbs

Marian Saniewski*l and Hiroshi Okubo*2

· 'Department of Plant Physiology , i-{eSf'nrch Inslitu Le of Pomology and Floriculture, Skicm ie'.'.ice, Poland

"'"Laboratory of HO lt kultnral Science, Faculty of Agriculture , Kyu shu University 46- 0 I , Fukuoka 812-8581, Japan

(Recei'ved Jury 2, 1998 and accepted August 7, 1.998j

The e ffect or phytotropin naphlllylpllt h<\ lanuc acid (NPA) , a well " UOWIl auxin transporL inhibito r, on tulip stem growth induced by indole-..'l--acelic add (IAA) in low (0.1%) and higll (2.0%) concentration in lanoliTl paste was sr.udied. IAA 0.1 and 2.0% applied on the p lace of removed Hower bud of sprout.ed precooled tulip bulbs and after f'!xds inn of aU leaves induced growt.h of all internodes; 1A.A. 0.1 % indllc~(l stem grO'.vth in higher degree I han 1AA 2.0% . NPA 0.2% applied on the 4th, :ln l alld 2nd internodes st.rongly inhibited the gwwt h of int.ern()d~~ illliuced by IAA 0.1 and 2,(/'16; Ihe inhibition was stronger in ca~p of IAA 0.1 than lAA 2.0 Ijf"). The inhibitory· effect of NPA on stem grow~h induced hy lAA was restored iJy add it. ional application of IA.-\ below NPA treatment. Tulip stem gl'o\<,th induced by !lower bud after excision of all leaves \ \-'a '; also inhibited by NPA be low its applicat ion . The inhihitory efrect of :"-J PA on stem growth promoted by 1M and t hat aoo it.ional nppHca tioll of IAA below NPA trea lment restored I he gTmvth illtlk ate on c rucial role of al lxin ill tulip stem gro\\t h.

INTRODU CTION

Tulip bulbs \\ith t.en ninal buds containing a complete flower require a period of 12- 14 weeks of low tempera LU re treatment for shoot e longation (De Herlogh, I ~J74). It is well kHOW II tha t elongation of the s tem and leaves of t ulips is due a lmost ent iTely t o the elongation of ceUs produced during ear lier developmental stages of flower bud formation (Gilford and Recs, 1973). The leaves and gynoecium provide auxins which control the elongation growth of the st ern (Op den Kelder et aI., 1971; Hanks and Rees, 1977 ; Saniewski and De MWlk, 198 1; Banasik and Sanicv./ski, 1985) . Saniewski and De Munk (1981) and Banasik an d Salliewski (HJS5) showed that elongation of all internodes in precooled tulip bulbs is promoted by application of auxins in the p lace of removed flower buds in the ahsence of leaves. Recently , it has been suggested that the elongation of all the inte rnodes in t ulips is con t rolled by the inte ract.ion of endogenous a uxins and gibberellins (Okubo a nd lJemoto , 1985, 1986; Okubo et at., 1986; Saniewski , 1989) . Okubo and lJemoto ( 1985) showed that 2,3,5-triiodobenzoic acid ermA) treatmen t at the first internode of sprouting tulip shoot inhibited the dark- induced elongation of thc first internode but did not affect the gibberellin amount. Recently, Saniewski and Okubo (1997) found that IAA applied in the place of removed flower bud and after excision of leaves promoted stalk e longation in the nonpl'eeo?led and precooled , rooted and derooled

*J. ~Co rrcspondeIH: e t o e it he r au t ho r . *lTe l: +48- 46- 33- 202 1, Fax: - 48- 46- :33- 3228, E- ma il : msanicw@i nsad .isk. s kiernicwice. pl "' .!Tp.l: +S I -02~642-2828 , Fax : +81· 92- 6-1 2- 2829, E- mail : ho ku bo@agr. k~ l IS h u- u. ac,j p

59

60 AI. Sanielt'ski and H. Okubo

tulip bulbs, and TlBA applied in the middle of the 4th internode (below L".A application) great.ly inhibited the growth of lower internodes. TIBA also inhibited tulip stem grovvth induced by IAA and NA.A.. in precooled rooted tulip bulbs after removal of flower bud and all leaves (Sanicwski and Okubo, 1998).

The phytotropin naphthylphthalamic acid (NPA) is known to inhibit polar auxin transport in dicotyledons and monocotyledons by inhihiting active auxin secretion (Depta et al., 1983), NPA inhibits auxin transport by specifically binding to the auxin efflux carrier (Katekar and Giessler, 1980). The endogenous auxin lAA does not compete vvith NPA for binding site (Lomax et al., 1995) and NPA-binding site is important for auxin transport (Ruegger fit al., 1997).

In this report the effect of NPA on tulip stem grmvt1l induced by auxins, IAA and NAA .. , in precooled rooted tulip bulbs is presented.

MATERIALS AND METHODS

Plant materials, temperature treatment and culture conditions Bulbs of tulip (Tulipa gesn.eriana 1.) cv. Gudoshnik, with circumference of

10-11 em, were used throughout the experiments. The bulbs, after lifting, \verc stored at 18-20 'C until 15 October, and then kept at 4°C for dry cooling. After cooling period, a minimum 12 weeks, the bulbs were planted individually in plastic pots and groV·,rIl in a greenhouse at 17 -20'C in natural light conditions.

Effects of NPA applied on the middle of the 4th internode on stem growth induced by IAA 0.1%

All leaves and flmver bud were removed when the lengt.h of the stem \-vas about 8 em and plain lanolin (control) or lanolin containing IAA at a concentration of 0.1 % was applied on the cut surface of the flower bud on 11 February. Then, 0.2% NPA in lanolin past.e \vas applied on the middle of the 4th internode and in other treatments additionally IA .... L\ 0.1 % was applied on the midcUe of the ;Jrd internode directly after NPA treatment or one day later after NPA application. Seven plants were used for each treatment. The grmvth of all internodes \vas measured every two days but that of 22 February is only presented.

Effects of NPA application on different internodes on stem growth induced by IAA at a concentration of 0.1 or 2.0%

IA.A at a coneentrat.ioll of 0.1 or 2.0% \vas applied on the top of the last internode after decapitation of flower bud and leaf excision as described above and then 0.2% NPA was applied in the middle of different internodes on 31 Januar~y. The stem length at. treatment was about 7 em. Sevcn plants were uscd per treatment. Measurement.s of different. internode grmvth were made every t\VO days but those of on 15 February arc presented.

Effects of NPA on the growth of stem of intact plants, growth induced by flower bud after excision of all leaves and growth induced by IAA

When the lengt.h of the stem was about 5 em 0.2% NPA was applied on the middle of

NPA on auxin-induced tulip stem gTOvAh 61

the 18t internode of intact plants or on t.he middle of the 3rd internode after removal of all leaves in the presence of flcnver bud. In the case \-vhen lAA 0.1 or 2.0% was applied on the top of the last internode after decapitation of llovver bud and excision of all leaves O.2°1r1 NPl\ \V(lS (lpplied on t.hp middle nf t.hp :ird int.prnnop. Seven plant.::> wr:rr: llSr:O pr:r treatment. Treatments were made on 6 March and measurements were done every two days during experiment period, but only the results on 17 March are presented.

RESULTS AND DiSCUSSION

Effects of NPA applied on the middle of the 4th internode on stem growth induced by IAA 0.1 %

IAA 0.1% applied in the place of removed flower bud of sprouted precooled tulip bulbs and after excision of all leaves induced grm'vth of all internodes (Table 1, Fig. 1). The results fully confinued previous studies (Saruewski and Dc Munk, 1981: Banasik and Saniewski, 1985; Saniewski, 1989), NPA 0.2% applied on the middle of the 4th internode inhibited t.he grmvth of all internodes induced by lAA 0.1% applied on the t.op of the last internode after decapitation of flower bud and removal of all leaves. The inhibitory efrect of NPA on tulip stem growth induced by IAA. was restored by additional application of IAA .. on t.he middle of the 3rd internode (helmv NPA) when applied directly or one day later after NPA treatment. It was found recently that iIl.hibitory effect of TIBA on tulip st.em growth induced by lAA was greatly restored by applicat.ion of IA6.. below TIBA treatment pla~e (Saniewski and Okubo, 1998). The grovv1:h of maize colcoptiles and pea internodes was greatl~yr suppressed when the organ was ringed "vith NPA and the grolNih suppressed by NPA was restored by IAA applied directly to the zone bclo\li,T NPA treatment (Morit.oshi Hno, personal communication).

Table 1. Effects of NPA (0.2%) applied on the middle oIthe 4th internode on stem grmvth induced by fAA (0.1%).

Length of internode (em) Treatment

lsL 2nd .1rd 4Lh

1. IntacL control 7.5 b~ G.O b 6.4 d 14.8 e dU lCQTJf~ iJJHl !1sYWfT bWl "ITPIQ1!t!d

2. Control (lanolin) 3.G a 1.6 a 1.1a 1.4 a 3. l;\.i\. 8.7 b 6.2 b G.G d 1:U1de 4. IAA + NPA (IV)' :3.8 a 2.0 a 19 b 9.9b 5. lAA + NPA (IV) + IAA (Ill) 8.2 b G.8 b 5.1 c 12.2 cd

6. TAA + NPA (IV) first, then lAA 011) 8.0 b S.H b 4.4 e 11.3 be one day lat.er

'"/I,·fean separation 'within columns by Duncan's multiple 'range test at 5% level. 'IV and ITT; NPA and IAA were applied on the middle of the 4th and :3rd internode, respecLively. InilialleJlgth at t.reatment: lst=2.8, 2nd= 13, :3rd:::O.H, 4th=1.2 em.

loud

34.7 d

7.7 a :35.2 d 17.G b :11.:3 cd 29.6 c

62 M. S'anielDski and H Okubo

Fig. 1. Bffecls of NPA ({).2%) applied on the middle of the 4th internode on stem grov.th induced by IAA.O.l%. 1-6: sec Tilble 1.

Effects of NPA application on different internodes on stem growth induced by IAA at a concentration of 0.1 or 2.0%

lAA 0.19'6 induced stem grmvLh of Lulip ill higher uegree LhaIlI1V" 2.0% (Tallie 2, Fig. 2). NPA applied on the middle of the 4th, 3rd and 2nd internodes greatly inhibited the grmvth of all internodes induced by 0,1 and 2.0% IAA, and the inhibitory effect of NPA was stronger in case of 0.1% t.han 2'()% IAA. NPA applied on the 1st. int.ernode did not inhihit. or only slightly inhibited the growth of the stern induced by either concentration of 1M.

NPA on au/t"if(-ind1.u:ed tuLip stern growth 63

Table 2. F:ffect.s of NPA (0.2%) application on different. in\.ern()(ies on stem grmvth induced by L\'A at a concentration of 0.1 or 2.0%

Length of internodc (cm)

Treatment 1st 2nd 3rd 4th

1. Intact control S.I e 0.7 I (i.4 g 18.8 g Allle'H·)f~ (lind tlQI~.'er· tnulnnnu!,.'ed 2. Control (lanolin) 3.2 a l.G a 1.0 a l.2 a

3. fAA 0.1% 8.9 f 6.7 f 7.1 h n:) f 4. [AA 0.1% + KPA (1\1' 3.9 ab 1.9 all ~.:l b 8.9 be .5. IAA 0.1% + NPA (III) 4.1 b 2.4 b 4,;3 cdc I 1.3 e G IAA 0.19'() + NPA eH) 4,:3 b 4.S (' fi.:i g 12.0 p

7. LA..;\ a. I 1M) + NPA (I) 7.1 d G.2 f 6.:3 g 12.4 pI

6. LA..;\ 2.0IJ& 7.3 d s.~~ e 4.9 def 10.1 d

9 lAA 2.01){1 ---'-- \.lPA (IV) 4.21) 2,\) c 4.1 e 8.6 b

III IAA 2.0% +NPA (TIl) 3.6 ab 3.0 c 4.2 cd n.o bed

11 IAJ\ 2JJ% + NPA (II) 3.8 ab :3.5 d 6.:3 f 8.7 be

12 lAA 2.0% + NPA (1) 6.3 c G.O e 5.0 ef 9.9 cd

'1-1ean separat.ioJ\ within colunms by Duncan's flIultiplp range test at 5')i') level. IV, III, II and!; l'<PA was applied on the miclcUe of the 4th, 3rd, 211(i aI\d 1st internode,

respectively. fnitiallength at treatment; Ist= 1.D, 2nd=1.2, ;jrd=O.7, 4th=LO em.

Fig. 2. Effects of NPA (0.2%) application on different intemodes on stem growth induced by TAA at a concentration of 0.1 or 2.0%.1-12; see Table 2,

Total

400 g

7.0 a 36.0 f 17.0 b 22.1 c 27.4 d :32.0 e 27.6 d ID.8 (" 19.8 c 2l.:1c 26.2 cl

64 M. Sanieu>.<;k'i and H. Okubo

Effects of NP A on the growth of stem of intact plants, growth induced by flower bud after excision of all leaves and growth induced by IAA

Tulip stem grmvth induced by flmver bud after excision of all leaves was also inhibited by NPA belovv its application (Table 3, Fig. 3). NPA applied helow the 1st node of intact plants only inhihited the growth of the 1 st interILode.

It is ,.,,'ell known that high concentration of IAA or NAA stimulates ethylene production in tulip stern and promoted internode elongation less and thickening more than 1m\' concentrations of these auxins do (Sanle\vski et aL, 1990). Silver thiosulphatc (8TS), an inhibitor of ethylene action, applied simultaneously \,>,ltlt high concentration of IAA or NAA greatly stimulated the stern gnHvth of tulip in comparison \vith r,he application of these auxins alone in the same concentrations (Saniewski et aL, 1990). Ethcphon greatly inhibited tulip stem grO\vth induced by flower bud or by IAA after removal of flower bud and all leaves and STS completely reversed the inhibitory~ effect of ethylene on stem elongation (Saniewski and Kavva, 1988). It. is possible that ethylene induced by high concentration of auxin itself inhibits polar transport of auxins via feedoack cOlltrol. 111 pea ;:;eedlillg.s v .. '1tefe pular auxin Lrflll~p()rt .i~ lll!ti1Jited by e1.llj-."jcne,

NPA binding sites were found to be reduced in number by the gas (Kang, 1987; Suttle, 1988).

The inhibitory action of NPA on tulip stem growth induced by auxin and the fact that lAA applied below t.he place of NPA treatment restored t.he grmvth indicate on the crucial role of auxin in tulip stem growth.

Table 3. Effects of NPA (0.2%) on the growt.h of stem of intact plants, growth induced by flower bud after excision of alilcaves and gro.,.;1h induced by IA.A...

-----_.-

Treatment. Length of internode (em)

1st 2nd ;·hd 4th

Intact plants

1. Control 8.0 e' 0.1 be RoO c 14.4 a 2. NPA (I)' 5.Sb G.G c R.O c 14.4 a AU lea"!)t'~ remQ!J@.d, tlQl"QQ.r bud. inlact 3. Control (lanolin) 6.1 b 5.6 b 8.3 c 16.7 h 4 NPA (III] 4.6 a 2.8 a 6.7b 18.5b jU lUQ1)e~ an.d. trQ'U){' r b.J.1(i r(J.mQ1HjJ).

fl Control (lanolin) LR 1.I 10 12 (; TAAO.l% 1O.5e 8.0d 10.4 d 17.4b 7. IIAA 0.1% +NPA (III) G.Sb 2.80. 5.1 a 17.0b R fAA 2.0% R.9d 0.4 (" RIc 18.0 a 9. JAA 2.0% +NPA (III) 4.4 a 2.5 a 5.8 a 12.4 a

':Mean separation within columns by Dlll1can's multiple range test at 5% level. ·'T and Ill; NPA \VdS applied on the middle of the 1st and 3rd internode, respectively. Init.iallength at treat.ment; lst=I.8, 2ncl=1.1, 3rd=1.0, 4Lh=1.2 em

Tot.al

36.6 e 34.7 be

~36.7 c :326 be

5.1 46.3 d 30.7 b ;i7.0 {' 24.6 a

NPA on an-d11-indnced tulip stern grow til

Fig. 3. Effects of NPA (0.29'(1) on the grmvt:h of stem of intact plants, grmvth induced by Dmver bud after excision of all leaves and grmvth induced by IAA. I-B; see Table 3.

ACKNOWLEDGME'IT

65

The stay of M. Sanie\vski at KYllShu University, Fllkuoka. Japan was supported by the invitation fellO\vship program for research in Japan, Japan Society for the Promotion of Science.

66 .J-J. Saniewski an4 H. Okubo

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Dc Hcrtogh, A 1974 Principles for forcing tulips, hyacinths, daffodils, Easter lilies an(i Dutch irises. Sci(mtia flmtic., 2: :31:3-;355

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Gilford, J. III and A. R Rees 1973 Growth of the tulip shooL Scicnlia HUTtic., 1: 14:3-156 Hanks, G. H. and A. It Hees 1977 Stem elongation in tulip and narcissus: the influellce of floral organs

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