A modified MS media for regeneration of direct explants and long term callus cultures of tomato.

Tatchell, S. and A. Binns

Eighteen tomato cultivars have been cultured and regenerated from leaf or other tissue types on modified Murishige and Skoog media (1 mg/l thiamine HC1, 0.5 mg/l nicotinic acid, 0.5 mg/l pyridoxine, 3 M IAA-aspartic acid, and 5 M zeatin riboside). This media was selected for optimal tomato regeneration after tests on comparable levels of NAA, IAA,IAA-alanine, IAA-phenylalanine, isopentenyl adenosine, zeatin, and kinetin. Direct explants from stem, leaf, pith, epicotyl and cortex, and hypocotyl tissues that have been cultured for over 18 months were regenerated. This medium produces bigh numbers of shoots on direct explants, and limited numbers of shoots on long term cultures. On long term cultues, all varieties tested have been successfully regenerated; however, passage number and time required on regeneration media differed among varieties.

Observations of long term cultures of UC82 pith and stem callus indicated small explants (2 - 3 mm) were more responsive to regeneration than larger explants. Some differences were observed between tissue types on specific cultivars but this was not true of all cultivars. In general, leaf explants gave the greatest number of plants per explant and root the least.

Table 1 shows the cultivars tested and the percent of direct explants that produced shoots after eight weeks on this modified MS medium. The number of plants that could be transferred to rooting medium at 8 weeks is also indicated. This number of ten increases significantly after an additional transfer. Those varieties that could regenerate shoots after long term culture are also indicated.

Table 1. Regeneration of direct explants and long term callus cultures of tomato.

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                             % direct explants
           Tissue    Callus  regenerated/number  Regenerability
Line/cross source(s) growth  plants per explant  long-term callus
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UC82       leaf     very good  98-100/2.6        good
           hypo     very good  95-100/NA         good
           root     very good
           stem     very good                    good
           epi & 
           cortex   good                         good
SP-l*      leaf**   good       >95/1.35          good
           hypo     good                         good
           stem     good                         fair
           epi & 
           cortex   good                         fair
SP-2*      leaf**   good       75-80/1.7         good
SP-4*      leaf**   good       >95/NA
LA13       leaf     very good  88-100/1.45-3.17  good
           hypo     very good
LA14       leaf     good       88-100/2.5-3      good
           hypo     good                         good
LA154      leaf**   good.      62.5/<l
LA159      leaf     good       >95/NA            good
LA274      leaf     very good  >95/NA            good
LA715      leaf     excellent
           hypo     excellent  75-90/2.8         good
LA811      stem     poor
           leaf     poor
           hypo     poor-fair  56-67/1.2         fair
LA1002     leaf     fair       75-80/NA
LA1223     leaf     good
           hypo**   good       95/NA
LA2363     leaf     excellent  98-100/4.38       good
Lukullus   leaf     good
           hypo     good       >95/NA
VFNT 
cherry     leaf**   poor       25/1.9            poor (1/76)
L. 
peruvianum stem     excellent  >95/>6            excellent
L.
chiloensis leaf     excellent  >95               excellent
           hypo     excellent  >98               excellent
           stem     excellent  >95/>6            excellent
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*SP lines provided courtesy of Dr. R.S. Poethig 
(SP-1 chlorophyll mutator;SP-2 e/e,u/u, wf/wf,mc/mc, 
SP-4 Xa2/+,a/a,h/h,t/t, and u/u).  **Sample size
for regeneration of these direct explants was less than 100.