indian
J.
Piant Physiol•• Vol. XXXIII, No.2. pp 113-118 dune. 199(})CYTOKININ ACTIVITY IN THE BLEEDING SAP OF DIFFERENT ROOTSTOCK COMBINATIONS OF ANAB-E-SHAHI GRAPE AND
ITS RELATION TO SOME GROWTH AND YIELD ATTRIBUTES
B.M.C. REDDY, G.S.R. MURTI AND K.L. CHADHN
Divisions of Fruit Crops. and Plant Physiology and Biochemistry, Indian Institute of Horticultural Research, Bangalore-560 089
Received on 24 July. 1989
SUMMARY
The cytokinin activity in the bleeding sap of'Anab-e-Shabi' grape grown on six different rootstocks was monitored for two years. The cultivar 'Gulabi' as rootstock and 'Anab-e-Shabi' on its own roots exhibited high rate of sap flow and cytokinin activity per unit time of sap flow. The rootstock 'Dogridge' which had a good sap flow rate, recorded low cytokinin activity per unit quantity of sap. The rootstocks, 'Teleki-5A' and '1616',
showed poor sap flow and bence found to have low cytokinin activity. The parameters such as days to bud burst, number of fruitful canes. and yield (ler ville showed a signifi cant positive correlation with the cytokinin activity of sap flowing per unit time. The predominant translocational forms of cytokinins tentatively identified from bleeding sap of 'Anab-e-Shahi' on its own roots are zeatin ribosidejdibydrozeatin riboside, and zeatin/ dihydrozeatin.
INTRODUCTION
The mecbanism by which rootstocks modify the development of scion varieties of fruit trees has been considerably discussed (~ogers :~nd Beakbane, 1957). The discovery that bleeding sap or xylem exudate contains both gibberellins (Phi1lips and Jones" 1964) and cytokinins (Loeffler and Van Overbeek, 1964) indicates that hormones produced by roots may play an important role in regulating metabolism in shoot of the plant (Kende and Sitton, 1967). It'is;~therefore, considered that hor mones produced in root may be involved in some of the effects of rootstocks on scion development (Luckwill and Whyte, 1968). Hence. it is probable that the bleeding sap/xylem exudate serves as a source of cytokinins in several plants. Such studies are of considerable importance in grape. which releases large amount of sap from its pruned ends. A number of rootstocks are available in grapes which are
li4
8.M.C.
iBnov
et ai.
capable of altering the growth and development of scion varieties (Winkler et ai.,
1974; Reddy, 1988). The cytokinins are reported to influence fruit bud diff~entiation in grapes (Srinivasan and Mullins, 198]).
In a rootstock trial marked differences in both vegetative growth and fruit yield between 'Anab-e-Shahi' on its own roots and 'Anab-e-Shahi' vines on five root stocks have been noted. It is possible that these effects may be related to the differential capabilities of the roots either to produce cytokinins or transport them to the top of the plant. A study was, therefore, undertaken to ascertain the effects of rootstocks on the flow of sap, its cytokinin activity and relationship with some developmental events and factors related to productivity.
MATERIALS AND METHODS
Investigations were carried out on grape cultivar 'Anab-e-Shahi' as scion with the rootstocks 'Gulabi', '1616', 'Dogridge', '1613', 'Teleki-5A' besides the one grow ing on its own roots. The bleeding sap was collected from tbe pruned cut ends into a test tube during October of the years 1983 and 1984. The duration of collection of the sap was recorded. Data were collected on number of days for bud burst and fruitful canes after October pruning during 1983 and 1984, while number of buds sprouting on the terminal 50 em long secondary was collected after April pruning during 1983 and 1984. Observations on bunches and yield per vine were recorded in February-March during 1984 and 1985.
Estimation of cytokinin activity from bleeding sap
10 ml of the crude sap obtained from each of the six root-stocks was directly incorporated into the bioassay medium and cytokinin activity assayed by Soybean cotyledon callus test (Miller, 1965) using cv. Acme. The medium described by Miller (1967) was used in the bioassay. The bioassays were conducted in 100 ml Erlenmeyer flasks using 50 ml of the medium. lhe number of explants per flask was three. The data are expressed as zeatin equivalents per hour of sap collection.
Purification of sap and analysis of eytokinins
115
equivalent to 40 ml sap was estimated using soybean cotyledon callus bioassay as described earlier. The data are expressed as mean fresh weight of individual pieces of callus.
RESULTS AND DISCUSSION
The means of the two years data. i.e. 1983 and 1984 pertaining to the rate of sap flow, cytokinin activity, days to bud burst, number of buds sprouting and fruit ful canes, are presented in Table-I. The means of the bunches and yield per vine for six rootstocks are for the years 1984 and 1985.
Sap How
The flow of sap was the highest with 'Gulabi' as the rootstock (6.64 ml/hr) followed by 'Anab-e-Shahi' on its own roots (4.55 ml/hr). The rootstocks 'Teleki 5A' (0.48 ml/hr) and '1616' (0.45 ml/hr) yielded extremely poor How of sap. The two rootstocks, 'Dogridge' (2.66 ml/hr) and '1613' (2.05 mJ/hr) were noted to be intermediate.
Cytokinin activity
The cytokinin activity expressed as zeatin equivalents/hr of sap flow was the highest in 'Gulabi' followed by 'Anab-e-Shahi' on its own roots. The activity was extremely low in the rootstocks 'Teleki-5A' and 'Dogridge' and was intermediate in '1613'. The cytokinin activity in "1616' could not be analysed because of negligible quantity of sap collected in one of the years. Inspite of a reasonably good rate of sap flow, the cytokinin activity in 'Dogridge' was poor because of low activity per unit quantity of sap.
Bud burst
'Gulabi' as rootstock hastened bud bursting closely followed by '1613' and 'Anab-e-Shahi'. Dud burst in the rest of the three rootstocks was considerably delayed. The numbet of buds sprouting on the terminal 50 cm long secondary after April pruning was highest in 'Anab-e-Shahi' on its own roots followed by 'Gulabi' and '1613'. It was low in 'Dogridge' and '1616'. The correlation between cytokinin activity per hour of sap flow of the rootstocks (excluding '1616') and days to bud burst were significant (r=0.8816). Some workers reported the presence of cytokinins in the bleeding sap of grape vines before bud burst (Nitsch and Nitsch, 1965; Srinivasan and Mullins, 1981).
Fruitful canes, bunches aDd yield per vine
..
-
0., Table I. Sap flow, cytokinin activity, bud burst and some yield attributes of 'Anab-e-Shabi' grown on different rootstocks Rootstocks 1. Anab-e-Shahi 2. Gulabi Rate of sap flow (mI/hr)4.55 6.64 Cytokinin activity
/hr
of
sap
zeatin
equivalents, (.L
moles) 0.2940 0.3650
Days
for
bud burst 22.23 17.88
Mean of 1983 and 1984 No. of buds Fruitful sprouting
canes! vine
20.82 59.03 18.70 83.64 Mean of 1984 and 1985 No. of Yield! bl.'!llchesl vine (kg)
vine 82.07
27.53
118.42
38.96
tit
k
p'
i
>< (i,-3. 1613 l2.05 0.1660 21.00 18.59 67.59 91.21 31.43
Q :---,~
~ 200 100
c.
" r 'CYTOItlNlN ACTlVtTY IN GRAPE SAP 117
SA' was intermediate while '1616' and 'Oogridge' were inferior with respect to the above characters. The correlations between cytokinin activity per hour of sap flow of the rootstocks (excluding '1616') and fruitfu1 canes (r=O.8674), bunches per vine (r=O.9217) and yield per vine (r=O.8855) were significant. These results point out a remarkable positive influence of cytokinins on the yield attributes. Cytokinins are implicated in the control of many aspects of grape reproduction including inflore scence formation (Srinivasan and Mullins, 1978; 1979), differentiation of flowers (Mullins, 1968), fruit set (Weaver et al., 1965), fruit development (Weaver and Van Overbeek, 1963). Based on the a\tailable information, they suggested that the endoge nous cytokinin is a primary regulator of reproductive growth in grapevine.
Cytokinins in the bleeding sap
Two compounds could be detected prominently from bleeding sap of 'Anab-e Shahi' on its own roots. These compounds eluted in fractions corresponding to zeatin riboside (ZR/dihydrozeatin riboside (OHZR) and zeatin (Z)/dihydrozeatin (DHZ) (Fig. 1). The cytokinin activity was more prominent in the fractions corres ponding to ZRjOHZR elution. It thus appears that ZRjDHZR and ZjDHZ are the predominant translocational forms of cytokinins in grapes. Several other workers reported that zeatin, zeatin riboside and zeatin ribotide are found in the xylem sap of grape vines before bud burst and during the period of active shoot growth (Loeffler
'"'
~
ZR Z lPA .i.P
a.
"
E '-"
'"
;..
i:i gOO
iii 800
~ ~ 700 ... ...
« 600
u
~ 500
Q
~ Aoo
8
300~
)
o
II')
ELUTION VOLUME (ml )
118
4Ii
B.w.e. RI!DDY
et al.and VanOverbeek, 1964; Nitseh and Nitsch, 1965; Skene, 1972; Skene and Antcliff, 1972).
REFERENCES
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Loeffler, J.E. and Van Overbeek, J. (1964). Kinin activity in coconut milk. In: Regu!ateurs naturels de la croissance vegetable. pp. 77-82 (J.P. Nitsch. Ed.) C.N.R.S., Paris. Luckwill, L.C. and Whyte, P. (1968). Hormones in the xylem sap of apple trees. In : Pial t growth
regulators pp. 87-101. Soc. Chem.lndust. Mong. No. 31. l.ondon.
Miller, C.O. (1965). Evidence for the natural occurrence of zeatin and derhatives: compounds from maize which promote cell division. proc. Natl. Acad. Sci., 54 : 1052-1058.
Miller, C.O. (1967). Zeatin and zeatin riboside from mycorrhizal fUngus. Science., 157 : 1055
1057.
Mullins, M.G. (1968). Regulation of inftorescence growth in cuttings of the grapevine (Vitis vint/era L.) I. Expt. Bot., 19 : 532-543.
Nitsch, J.P. and Nitsch, C. (1965). Presence de phytokinines et autre'> substances de croissance dans la seve d' Acer Saccharum et de Vitis vinifera. Bull. Soc. Bot. Fr., 112 : ll-P).
Philips, I.D.J. and Jones, R.L. (1964). Gibberellin-like activity in the bleeding sap of root systems of Helianthus annuus detected by a dwarf pea epicotyl assay. Planta, 63 : 269-278.
Raja Rao, T., Murti, G.S.R. and Challa, P. (1983). Cytokinins in gladiolus (Gladiolus grandi /iorus) corms. Ann. Bot., 52 : 703-710.
Reddy, B.M C. (1988,. Investigations on the effect of rootstocks in Anab-e-Shahi grape (Vitis vini/era L.). Ph. D. Thesis, University of Agricultural Sciences, Bangalore.
RogelS, W.S. and Beakbane, A.B. (1957). Stock and scion relations. Ann. Rev. PI. Physiol., 8 :
217-236.
Skene, K.G.M. (1972). Cytokinins in the xylem sap of grapevine canes: Changes in activity during cold-storage. Planta. 104 ; 89-92.
Skene, K.G.M. and Antcliff, A.J. (1972). A comparative study of cytokinin levels in bleeding sap of Vilis vini/era L. and the rootstocks, Salt Creek and 1613. J. Exp. Bot., 23 : 283-293.
Srinivasan, C. and Mullins, M.G. (1978). Control of flowering in the grapevine (Vilis vini/era L.):
Formation of inflorescence in vilro by isolated tendrils. Plant Physiol., 61 : 127-130.
Srinivasan, C. and Mullins, M.G. (1979). Flowering in Vilis: Conversion of tendrils into in florecences and bunches of grapes, Planta., 145 : 187-192.
Srinivasan, C. and Mullins, M.G. (1981). Fhy~iology offIowelil1g in the graJ(tvine-A review. Am. J. Enol. Vilic., 31 : 47-63.
Weaver, R.J. and Van Overbeck, J. (1963). Kinins stimulate grape growth. Calif. Agric., 17 :
12-13.
Weaver, R.J., Van Overbeck, J. and Pool, R.M. (1965). Induction of fruit set in Vilis vinifera L. by a kinin. Nature, 206 : 952-953.
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