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PNL Volume 19 1987 RESEARCH REPORTS
THE EFFECT OF VARIOUS SUBSTANCES ON IN VITRO ROOTING OF TWO PEA
GENOTYPESl
Cardi, ., E. Filippone C.N.R., C. S. Miglioramento Gentico
Ortaggi, Portici, Italy
and F. Adamo Cattedra Genetica Agraria
Universita di Napoli, Italy
In a previous note an effect on the genotype on in vitro
rooting of pea shoots was shown (1). In this report the effect of
various substances on in vitro rooting of two pea genotypes is
reported.
Shoots of 'Proteo' and L14 (afila) genotypes were obtained
either from excised buds cultured on P2 medium with BAP and IBA
added (2) or from seeds cultured on POb medium lacking hormones,
according to the technique described elsewhere (3). Shoots, 10-15
mm long, were excised and cultured on a rooting medium consisting
of MS salts half strength, sucrose 10 g/l, activated charcoal 2
g/l, CaCl2 220 mg/l, agar 8 g/l, pH 5.8 (2). The formation of
roots was scored after about one month of culture in a growth
chamber at 24C and 16h light.
Table 1 shows that when rooting was induced on the above-men-
tioned media, genotypic differences between Proteo and L14 were
significant only when shoots were obtained on the medium with hor-
mones. Although the presence of hormones in the proliferating
medium reduced rooting in both genotypes, the decrease was dif-
ferent, suggesting an interaction between hormone and genotype
effects. On the basis of results reported, it is not possible to
relate this effect to the af locus or to other loci. The compo-
sition of the rooting medium was modified as indicated in Table 2.
In these experiments all shoots were obtained from buds cultured
on P2 medium.
As concerns auxins, NAA 0.2 mg/l and IBA 0.4 mg/l gave the
best results. In peas, NAAl mkM was also found by Kartha et al. (4)
to be the best concentration. The absence of sucrose was always
detrimental, as reported elsewhere (7,8). In L14 a linear rela-
tionship between sucrose content and rooting percentage was evi-
dent. According to Maene and Debergh (5), the role of sucrose
seems to be more nutritional than osmotic. L-ornithine had a
strong negative- effect in both genotypes. On the other hand, 3-
aminopyridine significantly improved the response of both Proteo
and L14, confirming the results obtained in red clover (6).
1- Contribution no. 29 from Centro di Studio per il
Miglioramento Genetico degli Ortaggi-CNR, Portici, Italy
PNL Volume 19 1987 RESEARCH REPORTS
5
1. Cardi, T. and E. Filippone. 1986. PNL 18:7-9.
2. Filippone, E. 1985. PNL 17:12.
3. Filippone, E. and T. Cardi. 1986. Proc. Eucarpia Meeting on
Pea Breeding, Sorrento (Italy), June 10-13, 1985. pp. 71-85.
4. Kartha, . K., 0. L. Gamborg, and F. Constabel. 1974. Z.
Pflanzenphysiol. 72: 172-176.
5. Maene, L. and P. Debergh. 1985. Plant Cell Tissue Organ
Culture 5:23-33.
6. Phillips. G. C., and G. B. Collins. 1979. Crop Sci.
19:59-64.
7. Samartin, A., A. A. Vieitez, and E. Vieitez. 1986. J.
Amer. Soc. Hort. Sci. 61:113-120.
8. Travers, J. N., C. J. Starbuck, and N. J. Natarella. 1985.
HortScience 20:1051-1052.
Table 1. Rooting percentage in two pea genotypes on shoots obtained by
means of two different procedures.
Genotype
-------------- Shoots from-------:----------
buds on P2 medium
seeds on POb medium
Proteo
(330)1 54.8
(124) 93.5
L14 (afila)
(351) 34.8
(118) 90.7
X2
28.6 ***
0.3
*** Differences between genotypes significant at 0.001 level.
* In parentheses no. of shoots analyzed.
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PNL Volume 19 1987 RESEARCH REPORTS
Table 2. Effect of various substances added to the basal rooting medium on rooting percentage in
Proteo (a) and L14 (b) genotypes.
(a)
Level *
NAA
IBA
Sucrose
Ornithine
Glutami ne
3-aminopyridine
%
%
%
%
%
1
2
3
4
(58)2 55.2
(53) 71.7
(54) 63.0
(56) 64.3
(28) 46.4
(27) 55.6
(25) 64.0
(25) 56.0
(56) 39.3
(49) 61.2
(44) 68.2
(46) 65.2
(56) 55.4
(51) 17.6
(55) 7.3
(53) 1.9
(55) 52.7
(53) 64.2
(46) 67.4
(54) 70.4
(51) 52.9
(53) 83.0
(49) 77.6
(54) 77.8
X2
3.3
1.7
11.0*
58.2***
4.2
14.2**
(b)
Level
NAA
IBA
Sucrose
Ornithine
Glutamine
3-aminopyridine
%
%
%
%
%
%
1
2
3
4
(48) 33.3
(52) 61.5
(50) 58.0
(50) 50.0
(17) 35.3
(15) 60.0
(17) 70.6
(15) 60.0
(40) 22.5
(38) 31.6
(32) 46.9
(37) 51.4
(39) 30.8
(39) 20.5
(36) 8.3
(37) 2.7
(41) 39.0
(48) 50.0
(48) 60.4
(45) 57.8
(48) 33.3
(52) 71.2
(52) 69.2
(53) 71.7
X2
9.3
4.7
8.6*
13.4**
4.8
21.8***
*, **, *** Differences between levels significant at 0.05, 0.01, 0.001, respectively
1 NAA, IBA : 1= 0 mg/1; 2= 0.2 mg/l; 3= 0.4 mg/l; 4= 0.6 mg/l.
Sucrose : 1= 0 g/l; 2= 10 g/l; 3= 20 g/l; 4= 40 g/l.
Ornithine, Glutamine : 1= 0 g/1; 2= 0.5 g/1; 3= 1.0 g/1; 4= 1.5 g/1.
3-aminopyridine : 1= 0 mg/1; 2= 0.1 mg/1; 3= 0.5 mg/1; 4= 1.0 mg/1.
In parentheses no. of shoots analyzed
*****