Moss, J P

Nalini, H S

Sastri, D C

ICRISAT Open Access Repository

?f.AW TISSUE CULTVRE GENETIC MANIPULATION  AN^ 40bWTIC HYBRIDISATION OF PLANT CELLS e d i t e d  hy P . S .  Rao, M . R .  Hehle and M.S. C h a d t , ~  Proceedir~gs  of  National Symposium held at B h a b ~ . ~  A t o r ~ ~ i c  Research Centre Bombay, India 1980. IN  VlTRO CULTURE OF AMCHIS OVARIES AND OVULES. D.C.SASTRI. M.S.IULIN1 and J.?.MOSS Groundnut Improv-nt Progrvn ICRISAT. ICRISAT P a t a n c h r u  P.O. 502 $4. A.P.. INDIA. INTRODUCTION The c u l t l v a t c d  groundnut h y w a e a  L . )  I s  suscsptlbl.  t o  a number o f  d lseases and pests .  There cause a  cans lderab lc  loss I n  v l a l d  (1)  and t h i s  J u s t l f l a s  t h e  urgent  a t t m t ~ o n  of  groundnut b ~ o r o p ~ s t .  bu t  n o t  a11 tha rmsls tance genes h w a  been found I n  A.hypgaee. Fort,,. n a t e l y ,  I n  the w l l d  c o l l e c t t o n r  o f  t he  genus a number o f  species are  k n a m  t o  be r a s l s t a n t  t o   ma dlseases (2 ) .  I n  t he  genus Ar.chil there a re  s w a n  sections; hypogaea has bean crossad o n l y  w l t h  spec1.s from I t s  an s e c t l a ,  Arachls. though r m e  o t h e r  I n t c r s c c t l s n a l  hybr ldr  have baen produced ( 3 ) .  Ssme Inte.3.. 7 lona l  crosses for- pads, but N t U r e  seeds a r e  mt poss lb l a  due t c  s , r l y  degmera t l on  o f  cnd0sP.m. as I n  A.hypogaca x A . d l w o l  (41, o r  o f  embryo o r  both. n a b e r r  of t h  Sac t l an  Rhlzamatosae a ra  particularly v a l w b l e  f o r  d l s c r s e  and rmslst*nce. bu t  cannot be crossed w l t h  A.hypogaea. A proJ-c t  was s t o r t a d  I n  t h e  G:. , . .  2 . i t  Improv-t P r q - n m  at  ICRISAT t o  I n v e s t i g a t e  I n te r sec t  Ion,' i n c m p a t l b l l l t l a r  and t o  d*vel0: tachn lques t o  w e r c m r  them. E n c o ~ - .  :e b i  t he  repo r t s  of r ~ c c c . s f * ~  cu l t u rm  o f  w u l a r  a t  va r i ous  s t a g r  n' ot,elopmcnt I n  a  nwlber  of tu' (Tabla  1; to0 a l s o  6); we s t a r t e d  cult , :re o f  Arach ls  n m t l c o l *  O V J ~ ~ '  I n  v l t r o .  V. have a l s o  a t t m p t e d  t o  nlrml.ta p.9 m l m g a t l a n  w d  pd ~ n t r l b u t l o n  o f  t he  I n t e r n a t l u n a l  Crops Research I n s t l t u t .  f o r  tk *'- 1 Trop ics .  ICRISAT J,ur...' i r t  . c l e  A r p r w a l  k. 5 C - 3 .  ~ ~ ! t l . t l s n  I n  v l t r o .  There tw ln  o b J e c t t r t s  arc aimed at  oh ta l n lm)  f mll.blc mr thod Tor c m s s l n g  desirable bit ~ncanrat l b l e  stmcler of i. -. I n  t h l s  r epo r t  we presant  out a t t a m t ,  t o  c u l l u r e  011r1es and avu les  a: d l f f e r e n t  l n t e n a l s  m f t c r  p o l l l n a t l m ,  u l d  a l so  t c  pollinate m u l e s  , In r l t r o .  :c.  I IZ-d ovu les Gossyp lm  a r b o r e m  z v c - '  r s t a g e  x G.hlrsutVn t . h l r s u t l m  C h l r s u t m  Zn r h Ze~hy ran thes  sP b e t w n  15W avd 1/00 hrs  end p o l ' i  -an 0600 anu 'lESULTS -tha f o l l a l n g  day. Tha f l w r s  o r  5 ~ ~ -  - 3 ovaries +. i I l s c  , . ,hm~d ~ l l g h t  e ? - ~ a t ~ o r  2  - - - ' a n t  r f ' - -  w l t h  ch lo r lne  water, and ovules and L . ,  >r the ter 3 '  m o f  t ,CQTCI-  711 at ions - . I  . ~ a .  n &as . ~ L I  d 5 1 e d ,  2eqs dwelop lng  peg w r e  d l rsec ted  under a w c -  cond l t lons  , I r m s f e r r  * +  ,- . 7 ,ere no: 3 5  th-v e a r e d  ' .; And :, o A  to c u l t u r e  tub.¶ a t  d a l l y  I n t e r v m l l  f r a  !'.la d r y  o f  p o l l l n a t l a r  u n t l  ran t h e  t l p  ba tore  panet ra t  "3  'he 301 1 10 days a f t e r  POl l tnmtlon (Table 2 ) .  Ovules were e l m  p o l l l n e t e d  I n  v l t r o .  A mrl- of  l O f  of the m l l a  geminated  a n  - v i l e r  a r e  p o l l l -  2.  kt* o r  w d w a l o w m n t  a f t e r  PI I ~ n ~ t ~ m  - nated i -  " i f - o .  "any '6 -k . ~ e =  ,rw long but i e r  - -  "ere obsewed Dayr a f t e r  pol !  I n a t t o n  P q  Impth No. of C ~ ~ I S  No. or  O V L ~  . - t o  t~ 3v.' -=, t  growth r-' the war h e n  In m b r y o ( A q p m d  c u l t u r  'rey r s r e  w l t u r m d  and wll - > t a d  I n  v l t r o  on ht rasn lgc  ard Ska>p'r madlum I n  v l t r o  pol  l l n e t e d  134 uppl-ted v l t h  c a l e l n  h y d r 9 l ~ r r t e  6 0 0  mg/ l ) .  w i t h i n  2 - 3  w ~ + k ¶ ,  the Day of p o l l l n e t l a n  22 w l a r  grc* t o  5 t l m r  t h e i r  o r i g i n a l  size, I n  I 2  hrs I l g h t  a l t e m a t l n p  I - 3 days less  th.n 1-2 13 I t h  12 h r )  j a rk .  Later they kc- b r a n  end necrorlc, a t t a r  p o l l  l n q t l m  1.0 a 4 - 6  days 1-2 a 3-7 10 The m l e r  d l d  not r e r p n d  *rl l t o  othor media ( ~ a b l m  3 ) .  There a f t e r  pot I I M ~ I ~  7 - 10 d b p  3-4.5 Q 8-16 IZ was m l y  l rnarglnal I n c r w r e  i n  th. s i z e  of t b  m u l e s  which bwr b m  e f t a r  p o l l  l ~ t l o n  brr t h n  10 d a p  r l t h l n  threa m k s  and curd daveloplng l r r e ¶ p . c t l v e  of th. photocondl- 7-10 ca af te r  pol I m a t  ran I 2 b ( I n  ~ 1 1 )  I t l c n r .  h a  than ZW -lay o f  A.montlsole nn cu l tu rbd  at v a r l a r l  7-10 a I 12 (mt I n  w l f k  1 mom t h  l t w e r ,  but no t r d l l n g s  a w l d  k r r l d .  E l g h t v f w r  of there  w l a *  5-7 a L 16 Cln w l l )  1 33 I nr. t m r f e r r e d  t o  c u l t u r e  more thm 7 days a f t e r  ~ o l l l n . t l o n ,  l st.9. 5-7 a 11 (not I n  -11) L n t  a t &  ~ r t l n  ( I t )  reparted he s o r l d  recover " r a n  ~ w d l l n O 1  frm 74 owlu. A rml[.bla dlu, t e c h f g u *  *d fr.4tmrrt stlll For tk t n l t l b l  mxmrlnmmta t h  & l a  o f  h r e s h l g a  md SLrOOp (26) evelop.d kfora mutlnelT culmr. wuln f r m  1 n o r p . t I b l e  c*s*@. W l r a  (27). ~t.o md nfchay luk  08).  b r t l n  (12) md f l v  mnd Zmsky (29 * l t h  or * I t k t  m d l C f u t l m a  nra  u s d .  Table 3. Responses of  A.montiwla o v u l t s  t o  some rnedls Ovules HS MS w l t h  KF Hart  I n  Wrl:c w l t h  - IAAO n l g l l )  Kn I AA CH Fran NC S l  l gh t  H o c h a n g e , b u t  U n d e r t r l a l  HT N C NC I n  ovules p o l l  l n r t e d  swcl l l ng ;  ovules when t r a  pegs up f i a w a r l  runatn green t r r n s f e r r e d  frm t o  1L a long f o r  6 wcks KM show rmI I Ing then become fm NC a f t e r  Brown In NC N C NT N C  8 unpol l lnrtrd 9 weeks b ~ e k s  f l m r r  I I Pol I lnated Sl lgh t  I n l t l a )  S l i g h t  r m l l l n y  S w l l l n g  6 Very slow Sane o v u l e s  I n  vltro -- S w t l l l n g  s w l l l n g t h e n  thennochange  o c o r l o n a l l y  ce1' .15 form c a l l u s  I n  8 wecl. s no chanse, c a l l u s  I n  f o r -  ~ t l o n  brow I n  4 weeks j weeks AbbravI r t  Ions: CH Casein hydro lysate  HS R u r r s h l g e  anc Skoog'~  wdlum IAA Indo le  acet l c  r c l d  NC No change Kn K.o C M l c h y l u k ' s  d l u n  NT Not t r l e d  KN Klnet in ZZ  Z I v a l m s k y ' s ~ l u n  var los  Z lv  and Zm3ky could c u l t u r e  prps m d  Induce C.ud f o n w t l m  I n  rachia hyposaaa cu l tu red  m t  v a r l w s  stager o f  devmlopnont (19) .. h samm ~ d l t i o n s  of culturm, A.nontlcola pegs d l d  not respond. i i ~  urashlg- and Skoog's m d l u n  w i t h  k l n r t l n  ( 4 m g / l 1  1 1 2 ,  u i A r  (s. c l d l . ~ , ,  I i l  hQ.d p r o l l f l c  c a l l u s  I n l t i a t l o n  w t t h l n  a ru(; aftmr cu l tu re .  T h .  ~var l - s  whlch r-sponded h a t  r r -  tram fl-ra 2 and 1 d.ys a f t - r  p o l l l -  at lon, particularly vhon they bar- co ld  t l r a t e d  m t  bOc for 4 o r  l o r e  w r s .  I n  three r a k s  th- callurn rrr profusr  and a frr  org-tlc Iraas a u l d  b r  obsarvad i n  tho form o f  b o l b l l s .  Hand m d  m l c r o t o l .  mact lons ravamlad tha t  r y l r a  hd b6.n f o m d  and t b t  thr-  w n  r w l o n s  f i l c h  wore d l f f a r e n t l a t l n g  I n t o  r h m t s .  Xy lopmasls  war a r m  f r r q v r n t  rhrn such u l l u s  . n m  t r m s f r r r d  t o  U h l t a ' s  adlu r l t h  IM (4 q / l ) .  ,a of rhos* protukrmnces alongetmd t o  form pal*  g m u l  s t a l k  I I k e  irructurms which r m l n o d  organlrod but d l d  not d l f f - r m t l a t r  fu r rhc r .  uhan tha wgs wsrm cul tured on U h l t e ' s  nodlun r l t h  IM (*q/l). me culturmd p.9 showd m l n l t l a l  c a l l u s  a t  a rmqlon Just b l o w  t h  ~ v u l a s  and lacar  tha t l p  rhorod gootropism, tha l a t t e r  b o l w  t h  normal .aspens- of tho p.9 I n  v lvo.  A few o f  th. f o r m u l a t l ~ s  t r l o d  w f a r  hm supportad s m t h  o r  ca l lus f o m t l b n  whn u s d  t o  c u l t u r n  o w l e s ,  so our e f f o r t s  are ro I i r o c t - d  a t  t a s t i n g  th* m f f l u s y  o t  vmrlous w t s  I n t*  d l a  m d  a t h r  factors .  8. J . * .S tmar t  and C. I . d s u ,  J .tieredl r r  L ( 7 7 9 ) .  Arachls ovules do r e r w n d  b e t t e r  on a A l u m  u p p 1 r v l t . d  4 t h  ca ra ln  hydro1 -3te.  Careln f d m l y s a t e  r a m s  e s m t l a l  for  m I n l t l a l  g r a i t h  a r  has ' e o n  s h c n  f o r  ovules I n  Abelmorchus (11). Goacyplw (Zn a 417) \ : J t  N l c o t l n a  r u s t l c a  and U.tabacum (19). Haaver.  on a mdfum composed o f  HS n u c m l a m n t s ,  H a l l e r ' r  m l c m l a n t s ,  CvlIu'r v l t v n l n r  w i t h  k l n e t l n  m d  g l b b e r e l l l n ,  R a r t l n  (12) has abla t o  rear  r a m  r e d s  and seedl lngr  fmm A.hypogaea w u l e s .  Harcvar, the  b r t  g r a t h  o f  set fed A .mnt l co l8  o w l a r  occurred when the t l p  o f  t h  young gynophors was cul tured,  w l t b u t  e x c l s t l n g  th o v u l * ~ ,  on RS w l t h  caraln hydrolyrata ( 5 0 0  d l ) .  K l n a t l n  r s g u l a r l y  In&-d profus* c a l l u s  f o n r t l m  w I t h I n  t h r m  d a  when ycung gynophoras 2 and 3 d ~ y s  a t t a r  p o l l l n a t l m  w r a  cu l tu red  on llS w i t h  k l n e t l n  (%/I). A feu d l f f a r e n t l a t l n g  zmas c a r l d  br d e t c  ct .d krt t k r s  d i d  not  form shoots o r  roots. Only 3mt l i k e  s t ruc tu ra r  m r a  raan. h a .  cu l tu res  MY need t o  br t r m r f a r n d  t o  o t h r  m d l a  to contlnua d l f f e r e n t l a t l m .  To our Lmo*ldga, u l l u s  l n l t l a t l w r  from w a r l a r  m d  rubssquant rogmmrat lon o f  p lan ts  I n  I e g w s  t u s  kerr wuerar-  f u l  I n  on ly  au I n s t n u .  Wlth T r l f o l  lu p r a t m n  (31). REFEREMCES 1. II.U.Glbbon8. Dovalopmnt u ~ d  T r m s h r  o f  Techmloqy f o r  th. ra ln ted  a g r l a i l t u n  md th SAT f a r r r :  ICRlSAT (1979). 2. J ? s 8  hoe. tn t .  lagw Con?. bu (1978) ( I n  p m r r ) .  3. ~ . ? . G m r ) .  L W . C . 6 ~ ~ ~ k r a d l t y  185 (1979). 5. E.L.Johmsm L B.V.Snlth, k* r .J .b t .  bJ, 250 (19%). I. ~.kg$.n, ( n b r y o p m s l r  I n  v a s w l a r  p l m t r .  kd. . lc  Prmr. (1976). 6.M.S.hndIr. 8 0 t , C u . & 7 ( 1 9 7 2 ) .  7. C.A.Bmalq, B l o s c l u l a  906 (1971). 11. V.P.S.Bajaj. Proc. Vat1 I n r r .  Scl .  8 I J E B ,  175  (1364) 12. J.P.Hartln. OICa~insux u,  155 ( I ? : ' )  13. P.C.Jo¶hl. "Plant 5bryo logy :  A r y  rum Councll of 5' i c and I n d ~ ~ s t r l a i  Research N -151, pp 199 (19621 4 .  A.A.H.Eld. E.De.Lsnghs and L.Vaterkeyn, La Cc l lu le  6 9 ,  359 ( I : i ~ l .  5 .  C.A.8easley and I.P.Tlng. h r . J . B o t .  60. 130 (1973) 6 .  R.N.Chopra and P.S.Sabharwa1, ' P l a n t  t l t s u e  wrd organ w l t u r e  - A' svwoslum" (P.rahsshrar l  d M f .  9angawaq.ed) pp.257. l o?s rna t lona l  Soclety of  Plant * o r p b l o g l ~ t $ . D e l h l ( I % j ) .  7. H.Kuwada and T.Mbuchl .  Japan J . B r e d .  2 6 ,  298 (1376). 8. S.m.Read and G.B.Co,lliir, J .Yeredl ty  2, 311 (1978). 9. K.l(.nta and P.Mahe$hwarl. Phyranorphologr z, 210 (1%3), 10. N.#aheshuu.rl. Science x, 3hZ (1958). ! I .  H.Hahashrar1, and M.La1. Phy tmrpho logy  x. 107 (1961) 2 .  T.Uakizuka and T.Nakajlm., Japan J.lr.cd. 2. 182 (1974). : 3  T.'dakizuka and T.Nakajims. Japan J.Brtcd.  3, 161 (19751, .4.  F .L .Harner ,  h e r .  Potato, Jour. 1, 282 (1554). 5. T.Nalojlma, Y.Doymu and H.Eatsumoto, Japan J.Breed, 373 (1%)). 26. T.Hurashlga and F.Skccq, Phvslo logla P1 .2 ,  473 !1%1). 27. P.R.Vhlta, A nond book o f  P lan t  Tlssue Culture. J . C . t t a l l  ?raSS. M c a s t s r  (1943). 28. K.N.Km and M.R.Rlchayluk. Planta s, I05 (1975). 29. m.Zlv and E.Zamsky. Ann.lkt. 579 (1975). 30. P.C.Joah1 and B.tI.Johr1. PhyfomrphOlogy z, 1% (1972). 31. K.Horvath B u c h  and R.B.Snlth. Pl.Sclencr Le t te rs  & 231 Cl979). 

In Vitro Culture Of Arachis Ovaries And Ovules

Abstract

Similar works

Full text

Available Versions

ICRISAT Open Access Repository