Zero gravity effect on regeneration and normal growth of tobacco stem segment

Ball, E. A.

English

NASA Technical Reports Server

Semi-annual Status Report covering the period 1 October 1963 through 31 u c h c l ) -  
1964 on the research program entitled 1 
'3 GRCXITH OF A PLBNT TISSUE CULT= I N  T€E GRAVITY-FBE& STATE c-- Following the eqerimwts I reported by Skoog and Miller (19.59) it was Eroposed 
tp_gmr'as regenerative outpwths  from tobacco stem segments the'followiag oat+ 
gories of organs ortissues:  - 65- 32 0 9 0 
(ACCSSSION N U M B S R l  1, Normal shoots (stems with their attached 7 20 b o t s .  c 
(NASA C R  OR TMX O R  AD NUMBER) 
3. callus. I: 
since these organs or  tissues constitute the normal repertoire of the ' 
vegetative growth process i n  a nomal plant under the attraction of gravity, 
the experiments are considered important as a means of ascertaining the effect  
of the gravity-f'ree s ta te  upon the processes of regeneration and normal growth, 
The results of Skoog and lr8iller (1957) have been checked by continuing 
-- - 
- "-i- - _- __I_-- -- - I ~ .- ~ 
experiments; as these authors indicated, the various 
(production of the organs or tissues) generally occurs near the end of a perSod 
of time approximately 4.0 days after the Start of the expertimePts 
st ations of growth 
' 
. : -J  
The main problem of the present researches has been the accderation of the -~ 
growth responses; instead of obtaining these responses a t  the end of 4l dqys, $t 
is  essential to obtain them a t  the end of 20 days. 
2 
By utilizing an improved mineral supply in the culture medium for tobacw -?  - - I _  -- 
7 (htur-ge and Skoog, 1962) it has been possible to obtain the various required -.7 ,F - 
"2 
growth responses from stem segments, The accompanying photographs " show - --*. the * 3 
z 
r-3 
- L. 
tobacco stem segments d t h  stem plus lesves, apical callus, stem plus leaves 
and basal roots, roots alone, numerous buds without basal roots. 
w 
P= 
A3l of these pictured results have been achieved in l igh t  a t  oa. 10 foot oandles, 
It has not been possible to achieve these results in to ta l  darkness. However, since 
apparens l i gh t  requirement is of such small magnitude, it should not be too 
F 
d i f f i c u l t  tn n n d l f %  the exmrimntal set-tm in the sa te l l i t e  tn aacnmmndatrr it- 
https://ntrs.nasa.gov/search.jsp?R=19650022489 2020-03-17T00:59:51+00:00Z
. 2 
(I 
, Many supplementary experiments have been carried out  on the callus tissue, 
' a  
' since it w a s  the m a i n  point of interest by the writer i n  entering th i s  research 
program for the biosatellite. One of the most remarkable results achieved ia 
I_ - 
this program has been f r o m  the growth of the stem pieces in callus-producing 
culture medium rotated constantly on a klinostat. 
show a representative result Of such gro#t. 
The l a s t  three photographs 
Generally, the growth of the callus 
t issue op the klinostate is from 2 ta 3 tiroes as great (fresh a8ight) as that 
on the same medium not rotated. Other important differences are that the klinnstat 
callus is compect, while that f r o m  stationary mediun is  loose, friable; the kl3nnstat 
taus has such a high concentration of ohlorophyll in-the plestids that the whole 
qppears light green, Prhile that f r o m  stationary callus has 50 l i t t l e  chlorophyll 
that  it is almost w h i t e  (cf. figs. 2 anci 5). 
These diffemnces between the klinostat-grown and stationazpgrown callus 
are being investigated by l i g h t -  and electron microsco~y, and by deteminations 
of dry weight, and contents of protein, carbohydrate and fat. 
Brain (1939) suggested that the rotation of a shoot system during growth 
on a M i n o s k  b&s about a wnstatn redistribution of the auxin and caases a 
profound modification of the external and inter& growth patterns. No studies 
have yet been made to determine what growth factors ars behg redistributed 
by the constant reorientation of the callus by the klinostat, but the responses 
so far noted indicate others of more f u n d d t a l  aspect than a. 
Ref erenoes 
Brain, E. D. 19%. Studies in the effects of prolonged rotation of plants on 
a h o r i a n t a l  klinostat. 11. Anatomical structure. New Phytol. 38: 240-256. 
Idura&ge, To and F, Skoog. 1962; A revised mediun fo r  rapid growth &d bioassays 
i 
with tobacco tissue cultuzes. Physiol. Plant. 15: 4.73497, 
Skoog, F. and Go 0. EBiUer. 1957. Chemical regulation of growth and organ forma- 
. 
-- 
Figure 1. Tobacco s t e m  segment with a single large leafy shoot twenty days 
after inoculation on Murasbige, 20 g./l. sucrose and 0.2 mg./l. 
kinetin. The callus is of limited amount and quickly forms one 
or  more fast-growing buds. IX. 
. r--- ~ -I 
I - -  
I 
.I 
1-- .' 
k - - A  Figure 2. C a l l u s  of  tobacco stem with large t e rmina l  callus twenty days after 
inoculation on Murashige, 20 g./L sucrose, o.Z,mg./l. kinetin 
and 2 mg./l. indoleacetic acid. Top i s  the lateral view, and 
bottom is v i e w  from above. Note the friable nature, and the almost 
uhite color of this callus. UI. I 
c 
5 
. 
-- t 
. 
Figure 3. Tobacco stem segment with a conspicuous callus and a large upright 
leafy shoot twenty days after inoculation on Murashige, 20 g./l 
sucrose, 0.2 mg./l. kinetin, 2 mg./l. IU, and W O  
A s m a l l  adventitious root is seen a t  the base of the shoot. 
and/or the auxin greatly stimulates growth in length of the shoot as 
compared with figure 1. 
a t  4.00 mg./l, 4 
The phosphate 
6 
\ L  
. \  . 
Figure 4. Adventitious roots from t i p  of stem segment of tobacco inoculated 20 
days previously on Murashige, 20 g./l. sucrose, 0.02 mg./l. kinetin, 
and 2 mg./l. IU. The relatively high concentration of kinetin with 
respect t o  the one of IAA has brought forth a prolific rooting response, 
with no formation of buds. M. 
e 
' . -..... 
/-I 
.-- - - 
I 
t i  
I 
I .  
7 
I- 
- 
Figure 5. Calli grown from tobacco stem pieces inoculated 20 days p32eviously on 
Uurashige, 20 g./L sucrose, 150 nil& coconut m i l k ,  0.5 mg./1. 2,4-D. 
Upper l e f t  i s  top view, showing original diameter of stem Fnoculum, 
upper r i g h t  is bottom view that was i n  contact with agar medium, and 
lower photo is  la teral  view of the callus. T h i s  c&ius is of firm 
consistency, and l i g h t  green i n  color; c;ompare with fig. 2. U. 


Growth of a plant tissue culture in the gravity-free state semiannual status report, 1 oct. 1963 - 31 mar. 1964

Abstract

Similar works

Full text

Available Versions

NASA Technical Reports Server