Pak. J. Bot., 38(3): 603-612, 2006.

EFFECT OF STIONIC COMBINATION ON THE GROWTH

AND YIELD OF KINNOW MANDARIN
(CITRUS RETICULATA BLANCO)
WAQAR AHMED, M.A. PERVEZ, M. AMJAD, M. KHALID*, C. M. AYYUB
AND M. AZHER NAWAZ

Institute of Horticultural Sciences, University of Agriculture, Faisalabad.

*
Institute of Soil and Environmental Sciences, University of Agriculture, Faisalabad.
waqar199@hotmail.com

Abstract

Citrus is a leading fruit crop of Pakistan. Among citrus cultivars, Kinnow is the most
prominent and shares about 95% of total citrus production in Pakistan. The monopolized cultivation
of Kinnow scion grafted over rough lemon needs a substituted rootstock for higher yield/return and
for longer productive tree life. Trials for the selection of suitable rootstock for Kinnow mandarin
under subtropical environmental conditions and highly alkaline rhizosphere were carried out in the
experimental fruit orchard of the Institute of Horticultural Sciences, University of Agriculture,
Faisalabad during 2003-2005. The studies revealed that Volkamer lemon, Brazilian sour orange and
Citrumello 4475 were the reliable rootstocks for citriculture industry of the Punjab province.

Introduction

The importance of citrus to agriculture and worlds economy is demonstrated by its

wide distribution and large-scale production. Citrus is an important genus of the family
Rutaceae in the plant kingdom. Various species of Citrus are believed to be native to
tropical and sub-tropical regions of Asia and Malaya archipelago (Hooker, 1872). It is an
economically potential fruit and is grown in more than 52 countries of the world.
Pakistan is bestowed with suitable climate and soil conditions to grow citrus; where it is
grown on an area of 185,400 ha with annual production of 1.67 tones (Anon., 2005).
Today Pakistan stands among the top 14 citrus growing countries of the world.
There is no controversy over the importance of citrus rootstock for better citrus
production. A successful rootstock should have compatibility between scion and
rootstock besides having tolerance against prevalent edaphic and environmental
conditions. Rootstocks provide growers with useful tool to manipulate the vigor and
production of orchard trees. Effects on tree size, fruit quality, precocity, fruit production
and maturity are achieved through complex interrelationship between the roots and
canopy of the plant. Castle et al., (1988) reported that trees on Trifoliate orange and
Carrizo citrange gave the better performance with respect to total soil soluble and the
lowest TSS was gained from the trees grafted on Rough lemon. Constantin et al., (1979)
found that rootstocks significantly influenced the yield, fruit weight, rind thickness, peel
colour, juice contents, total soluble solids and titratable acidity of Owari Satsuma
mandarin when grafted on 16 different rootstocks. Similarly, many workers reported the
higher yields of sweet orange on Rough lemon root stock than on other rootstocks
(Brawn, 1920; Gardner & Horanic, 1961; Cohen & Reitz, 1963).
Rootstocks directly affect the ability of plants to take up water and nutrients and
significantly alter the pattern of canopy development and photosynthesis (Richardson et
al., 2003). Kinnow mandarin is inclined to progressive decline these days in Pakistan
604 WAQAR AHMED ET AL.,

due to abiotic stresses viz-a-viz shortage of irrigation water, dry weather and ever
increasing salinity in the root zone along with adverse impact of intercultural practices in
the orchards. An unusual important feature of our citrus plantation is the short life span of
trees which seldom exceed 25 years. In over 40% cases the decline of trees starts at the
age of 10 years which is the prime age of production, whereas it is not uncommon for
citrus orchards to remain in production for over 50 years in many other countries. Low
productivity and short span of productive life of citrus trees coupled with the foot rot and
excessive leaf drop along with twig drying are researchable problems under the adverse
environmental conditions. One of the hypotheses directs towards the incompatibility of
Rough lemon to Kinnow mandarin under highly alkaline soils of the Punjab province.
Among various factors responsible for citrus decline, rootstocks have been considered to
be a major contributor with respect to climatic conditions. Today rootstocks related
problem in Citriculture has assumed a great significance. Every citrus growing country is
faced with this burning problem and is trying to solve it. The rootstock found suitable at
one time, may entirely fail in future and that seems to be an original problem of the
Punjab province. One of the basic purposes of the present study was to find rootstocks
resistant to citrus blight. There is no single rootstock available that can be regarded as an
ideal for Kinnow mandarin cultivar under all types of agro-climatic conditions. Therefore
selection of proper rootstock for scion variety may lead to the failure and/or success of
the orchard. The present project was hence designed as a preliminary effort to investigate
a suitable rootstock for Kinnow considering vigour, productivity and fruit quality. The
citriculture industry of the Pakistan will have to address all such issues to make the citrus
industry sound and internationally competitive, particularly in fresh fruit export market.

Material and Methods

The studies were carried out at the Experimental Fruit Garden Sq. # 9 of the Institute
of Horticultural Sciences, University of Agriculture, Faisalabad. Nine rootstocks
Citrumello 4475, Citrumello 1452, Volkamariana, Yuma citrange, Rough lemon, Mithi,
Troyer citrange, Carrizo citrange and Brazilian sour orange were used.
For the physiochemical analysis, 100 fruits of each cultivar were picked during the
month of January for all the consecutive years of the study. The analyses of fruit were
conducted in Post-Graduate Pomology Laboratory, Institute of Horticultural Sciences,
University of Agriculture, Faisalabad. The following growth and physiochemical
parameters were studied on the plants: Scion girth, rootstock girth, canopy spread, plant
height, number of fruits per plant, Juice %, total soluble solids (TSS), acidity % and
vitamin-C contents in the juice. Girth measurement was taken at a fixed height of 15 cm
above the graft union for scion girth and 15 cm below the graft union for girth of
rootstock. The positions were marked with black paint for recurrent observations. Trunk
diameters were taken annually in the March, so as to quantify any differential growth
rates during the study period. Plant height was measured by telescope pole while canopy
spread was calculated using the following formula after measuring the outer peripheral
branches of the canopy in East-West and North-South directions.

Canopy spread = 4/3 r3.

Total soluble solids were measured with digital refractometer (ATAGO, RX 5000).
Acidity in juice was determined by taking 10 ml of juice from each sample and diluting
with distilled water in a 100 ml beaker. 2-3 drops of phenolphthalein were added for end
EFFECT OF STIONIC COMBINATION ON THE KINNOW MANDARIN 605

point. The samples were titrated against N/10 NaOH (Hortwitz, 1960). The results were
expressed as percent citric acid

N/10 NaOH used 0.0064

= ----------------------------------------- 100
Volume or Weight of sample used

For the determination of vitamin-C (ascorbic acid) in juice, the method described by
Ruck (1961) was used. Ten ml juice was taken into 250 ml conical flask and volume was
made up to the mark using 0.4 % Oxalic acid solution. Five ml of filtered aliquot was
taken in a flask and titrated against 2, 6, dichlorophenoindophenol dye to a light pink
colour which persisted for 10-15 seconds.

Vitamin-C was calculated as:

1 R1 V
= ---------------- 100 mg ascorbic acid per 100 ml juice
R W V1
where
R1 = ml dye used in titration of aliquot
R = ml dye used in titration of 1 ml of standard ascorbic acid solution prepared by
adding 1 ml of 0.l % ascorbic acid + 1.5 ml of 0.4 % oxalic acid.
V1 = ml of juice used.
V = Volume of aliquot made by addition of 0.4 % Oxalic Acid.
W = ml. of aliquot used for titration.
TSS/acid ratio was calculated by dividing the TSS over acidity

Statistical analysis of data

The experiment was laid out in a Randomized Complete Block Design (RCBD). All
data were analyzed statistically using analysis of variance techniques (Steel et al. 1996)
and the means were separated by Duncans multiple range test.

Results

Vegetative performance: Rootstock and scion girth is considered very important to

determine the degree compatibility of stionic relationship between stock and scion.
Sometime rootstock shows incompatibility with scion leading to unbalancing in
physiological functions, plant vigor, productivity as well as fruit quality. The
observations for this parameter showed significant differences among rootstocks for
increment in stock girth. Average maximum rootstock increment over the years was
observed in Rough lemon (2.00 cm) followed by Volkamer lemon (1.25 cm) whereas,
minimum in Brazilian Sour Orange (BSO) and Mithi showed almost no increment in
girth (Table1). The annual increment in girth of all other rootstocks was moderate and
remained statistically at par with each other. It showed slower rate of plant growth and
this trend depicted that Rough lemon is the most adapted rootstock under highly alkaline
soil of the Punjab province, while Volkamer lemon which is a sister rootstock to Rough
lemon is also well adapted to alkaline soil conditions and would be a future substitute of
Rough lemon rootstock.
606 WAQAR AHMED ET AL.,
EFFECT OF STIONIC COMBINATION ON THE KINNOW MANDARIN 607

As far as scion girth is concerned, the results indicated that Rough lemon had
maximum increment (3 cm) followed by Yuma citrange (2.50 cm). Volkamer lemon had
an average annual scion increment of 1:00 cm and rubbed its shoulder with Troyer
citrange and Carrizo citrange. Minimum annual increment in scion girth was observed
(0.75 cm) for Citrumello 1452, Mithi and Brazilian sour (Table 1). Degree of variability
in the compatibility of graft combinations has been observed on the basis of differential
growth rates of scion and rootstock i.e., even growth between Rough lemon rootstock and
Kinnow scion (Fig. 1). In another case Troyer citrange rootstock had over growth on the
Kinnow scion (Fig. 2) while in the stionic combination of Brazillian sour orange (BSO)
and Kinnow mandarin, scion had over growth on the rootstock (Fig. 3).
Similar diameter of scion and stock is an ideal indication of congenial relationship
which produces a reasonable crop of quantity and quality. It was noticed that Rough
lemon was at the top for the induction of increment for scion and stock. Trees on
Volkamer lemon were vigorous and had strong union followed by Rough lemon.
Vegetative growth has a profound impact on the reproductive growth of citrus
because citrus bears on current season growth emerging from one year old branches. So
keeping in view the bearing habit, canopy spread was included as a parameter of study.
Data was recorded and canopy increment proved to be non-significant as affected by
various rootstocks. However, maximum increment in canopy was observed in Citrumello
4475 (1.60 m3) followed by Volkamariana rootstocks (0.43 m3) while a minimum canopy
spread was observed in Mithi (0.01 m3).
Plant height is another important vegetative characteristic of citrus tree. Rough
lemon gained a height of (0.53 m) and proved to be superior closely followed by
Volkamariana (0.38 m) and Citrumello 4475 (0.31 m), while the minimum increment in
plant height was attained by Brazilian sour orange (0.11 m). Volkamariana proved as a
very suitable rootstock as it had incorporated vigour to the Kinnow scion.
The results for total number of fruits per plant (yield) indicated a significant difference
among rootstock treatments. It was found that Brazilian sour orange had maximum number of
fruits (1050.25) closely followed by Volkamer lemon (920.50) and Rough lemon (723.50).
Citranges {Troyer (411.00), Yuma (230.50) and Carrizo (155.50)} revealed poor results and
Carrizo Citrange (155.50) proved to be the poorest of all. Volkamariana gave bigger sized
marketable fruits as compared to Brazilian sour orange; which although produced more fruits
but their size was small so Volkamer lemon proved again as a potential candidate to replace
Rough lemon as far as yield component is concerned.
Physiochemicl analysis of fruits: Juice percentage in the citrus fruit is considered to be very
important. The ultimate demand of customer is higher juice percentage in the fruit. Results
regarding the juice percentage revealed a significant difference among different rootstocks.
Volkamariana was found to be superior and produced maximum juice percentage (50.84),
which was statistically at par with Rough lemon (49.92%) and Mithi (49.12%) followed by
Carrizo Citrange (47.33%), Brazilian sour orange (46.73%) and Yuma Citrange (46.51%).
Minimum juice %age was recorded in the fruits harvested from the trees grafted on
Citrumello 1452 (39.31%). Juice recovery is a qualitative parameter and Volkamer lemon
proved itself as a meritorious rootstock.
Data pertaining to TSS revealed significant differences among rootstocks. Maximum
total soluble solids were recorded in Mithi (13.57), which was statistically similar to all other
rootstocks except Yuma Citrange (11.44), which produced the minimum total soluble solids.
Other rootstocks viz., Brazilian Sour orange (13.33), Citrumello 4475 (13.40), Troyer citrange
(13.24), Citrumello 1452 (12.48), Carrizo citrange (12.44), Volkamariana (12.19) and Rough
lemon (11.84) were statistically at par with respect to total soluble solids (Table 2).
608 WAQAR AHMED ET AL.,

Fig. 1. Rough lemon-even Bud union configuration Fig. 2. Overgrowth of Troyer citrange rootstock
with Kinnow scion on Kinnow scion

Fig. 3. Overgrowth of Kinnow scion on Fig. 4. Vigorous Volkamer lemon tree

Brazilian sour orange rootstock
The data pertaining to acidity showed a significant effect of treatments on acidity
(Table 2). Citrumello 4475 had the maximum acidity (1.13%), which was statistically at
par with the Citrumello 1452 (1.02%), Yuma Citrange (1.09%), Carrizo Citrange
(1.02%), Troyer Citrange (1.02%) and Brazilian sour orange (1.14%). Minimum acidity
(0.88%) was recorded in fruits of Rough lemon. The chemical analysis was carried out
during January at the peak fruit maturity of Kinnow mandarin during the consecutive
years.
EFFECT OF STIONIC COMBINATION ON THE KINNOW MANDARIN 609
610 WAQAR AHMED ET AL.,

Citrus is believed to be the efficient source of Vit. C, which is a powerful

antioxidant. Statistically significant differences were observed among the rootstocks for
vitamin C contents. The data revealed that maximum vitamin C (33.90 mg) was recorded
for trees grafted on Rough lemon and Yuma Citrange followed by Carrizo Citrange
(33.74 mg), Mithi (33.45 mg), Volkamariana (33.43 mg) and Citrumello 1452 (32.98
mg). Minimum vitamin-C was recorded for trees on Citrumello 4475 (29.94 mg).
Mean values of the TSS / acid ratio showed a maximum ratio in the Mithi rootstock
(14.13) which is statistically significant and close to the Rough lemon rootstock (13.45)
while different to all other rootstocks. Minimum value of TSS / acid ratio was found
(10.49) in Yuma citrange, which is at par with the Brazilian sour orange (11.69) and
Citrumello 4475 rootstock (11.85).

Discussion

Rootstocks and scion girth equilibrium is very important for the compatibility of
rootstocks with the scion. Sometimes rootstock shows incompatibility with scion. In the
present study rootstock cause unbalancing in physiological functioning, plant vigor,
productivity as well as fruit quality. Stock and scion stem girth was significantly affected
by the various rootstocks under study. Although it has been demonstrated by earlier work
that rootstock has a profound impact on the vegetative behaviour of the scion depending
upon the compatibility; even then variation in the impact of rootstock on scion is beyond
the compatibility and it is also influenced by the environmental stresses; weaker
rootstocks become more weaker as in this case citrange rootstocks incorporated very little
vigour to the Kinnow scion and produced less yield with poor quality of fruits. Vigours
rootstocks like Rough lemon and Volkamer lemon performed equally well along with
Brazilian sour orange. Maximum rootstock girth was observed in Rough Lemon (2.0 cm)
followed by Volkamer lemon (1.25). This vigorous increment of the girth shows the
compatibility of Rough lemon and Volkamer lemon with Kinnow scion. This trend
showed that these rootstocks are physiologically compatible and actively growing under
the alkaline soil conditions as already reported by the Castle et al., (1988) who described
that Volkamer lemon is resistant to Phytophthora and alkaline soil conditions.
The vegetative performance of Volkamer lemon was comparatively better in terms of
plant spread and plant height (Fig .4). So it could be inferred that Volkamer lemon is a
promising rootstock for Kinnow mandarin under the dry climate of the Punjab province.
Environmental influences on the performance of rootstocks has been described by Moeen
(2000) and Noda et al., (2001), but the variability is mainly due to weathering agents and
intercultural practices on the plants. The climate of the Punjab province is very harsh
during summer season and vigorous rootstock is required to sustain the spring flushes. So
according to these studies Volkamer lemon proved quite satisfactory and could be used as
a substitute rootstock for existing Rough lemon as rootstock in Citriculture industry of
Pakistan.
Plant canopy development pattern is very much in line with the increment in stem
girth and plant height of various rootstocks under study. Kinnow Mandarin has a natural
tendency of compact canopy and this character is more or less maintained on all types of
rootstocks in the study. Volkamariana proved to be consistent in the increment of canopy
spread as against all other rootstocks whereby their response was quite inconsistent
during the course of studies. Our results are in agreement with those of Kumar et al.,
(1994) who reported that under arid environmental conditions, vigorous rootstocks are
required to give a boost to citrus plants.
EFFECT OF STIONIC COMBINATION ON THE KINNOW MANDARIN 611

Fruit yield is of prime concern to orchardists. They grow the plants for better yield
and good quality fruit production. Although the Brazilian sour orange produced much
higher number of fruits with well blended quality traits, the smaller sized fruits were not
of marketable grade. Volkamer lemon produced good quality marketable sized moderate
number of fruits and proved itself as a reliable rootstock for Kinnow mandarin. Our
results were closely related to those of Georgiou (2000) who also observed maximum
yield from the trees on Brazilian sour orange (BSO) and Volkamariana rootstock.
Volkamariana proved better as far qualitative characteristics are concerned.
Maximum juice percentage (50.84) was observed in the fruits on Volkamer lemon while
the Rough lemon contained 49.92% juice and the other rootstocks proved inferior for this
parameter of study. Granulation/riciness was also observed in the juice vesicles of fruits
harvested from Rough lemon and all the three citrange rootstock, whereas no granulation
/ riciness was observed in the fruits harvested from Volkamer lemon stock. It could be the
reason of more juice recovery in fruits on Volkamariana rootstock. Volkamariana proved
better in terms of qualitative characteristics. Recovery of high juice contents in
Volkamariana again showed that it could be used as a substitute rootstock.
Kinnow is considered to be full of vitamin-C and other elements like Mg, Fe and Ca.
Total soluble solids (TSS), acidity and ascorbic acid have their importance in
standardizing the citrus fruit quality parameters. The excellent performance of Bazillion
sour orange is due to inherent characteristics of sour orange. Across the world various
strains of sour orange are extensively used because of qualitative traits, although its
cultivation is banned in various countries due to its susceptibility to citrus tristeza virus
(CTV). Even then it could be inferred from the results that Volkamariana rubs its
shoulder with Bazillion sour orange for this trait and is equally good for its commercial
cultivation.
According to the results, there was a negligible impact of rootstock on acidity
percentage; but still it is valid information to make a baseline for the selection of
perspective rootstock. Although acidity is graft transmissible factor and affected by
rootstocks as determined by Castle et al. (1988), it is highly regulated by environmental
conditions. Homogeneous mildly cool environment during fruit ripening could be
attributed to negligible influence of rootstocks for this parameter of study.
According to the results presented here the impact of Vit. C assimilation is graft
transmissible and thus the rootstocks like citrange and Brazilian sour orange have
moderate amount of Vit. C as against Rough lemon, Volkamer lemon and Mithi (Table
2). Our observations regarding TSS, acidity and Vitamin-C were in consonance with the
findings of Jianguo et al., (2001) and Wutscher & Hill (1995) that rootstocks affect TSS,
acidity and Vitamin-C significantly.
Total soluble solids / acid ratio is affected by the various rootstocks. As shown in the
results, maximum TSS / acid ratio was recorded in Mithi and minimum ratio was
observed in Yuma citrange (Table 2). As TSS / acid ratio is a flavoring factor, so these
results depicted that with increase in the ratio there was a decrease in the acidity so with
low TSS / acid ratio, quality of fruit is poor and taste of fruit becomes watery and insipid.
Again the ratio is used to determine the fruit maturity standards, so where the ratio is
high, fruit will mature earlier. Zekri (2000) reported that higher the Brix: acid ratio the
earlier is the fruit maturity.
The results showed that although Rough lemon has meritorious characteristics for its
commercial cultivation, the susceptibility of Rough lemon to even weaker strains of
Phytophthora is a demerit and a cause of low productivity, shortened age of plant, early
tree decline and poor quality of fruits. The situation of Phytophthora attack aggravated
even more on Rough lemon under high alkaline soil conditions with less availability of
612 WAQAR AHMED ET AL.,

irrigational water. In contrast, Volkamer lemon was comparatively resistant to

Phytophthora (Castle et al., 1988) and as a vigorous rootstock suitable to our agro-
climatic conditions. Further trials are underway in this regard to certify the performance
of various rootstocks and particularly the perspective rootstock Volkamer lemon,
Brazilian sour orange and Citrumello 4475 for the widening of choice of rootstock uses in
Citriculture industry of Pakistan.

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(Received for publication 7 June 2006)