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All content following this page was uploaded by Houcheng Liu on 28 August 2014.
Received February 11, 2013 / Revised April 26, 2013 / Accepted May 3, 2013
GKorean Society for Horticultural Science and Springer 2013
Abstract. The effect of phosphorus (P) nutrition on plant growth and pigment formation in the flower stalk was studied
under hydroponic conditions for 2 Chinese kale (Brassica alboglabra Bailey) cultivars: ‘Jianyexia’ (green flower stalk)
and ‘Hongjiao’ (mauve flower stalk). Three different P treatments were used: 30 (normal-P), 7.5 (low-P), and 0 mg·L-1
(P-deficient). The results showed that the biomass, yield, plant height, stem diameter, and leaf number of Chinese kale
were significantly reduced in the low-P and P-deficient treatments compared to the normal-P treatment. The chlorophyll
content in the flower stalk epidermis was not affected by different P levels in ‘Jianyexia’, but was significantly
reduced by the P-deficient treatment in ‘Hongjiao’. Decreased P levels caused the flavonoid, soluble phenol, and
anthocyanin content of the flower stalks to gradually increase in both Chinese kale cultivars. The pH value of the
flower stalk epidermis gradually decreased with the declining P levels, and was significantly different among the 3
treatments. As the P levels declined, phenylalanine ammonia-lyase (PAL) and chalcone isomerase (CHI) activities in
the flower stalk epidermis gradually increased, and were significantly different among the 3 treatments. P nutrition may
control the synthesis of anthocyanins in the flower stalk by regulating the epidermal pH value, and the activities of
PAL and CHI.
Additional key words: anthocyanins, chalcone isomerase, phenylalanine ammonia-lyase
was also affected by the presence of other pigments, such as (1976). 0.5 g sample of flower stalk epidermis was extracted
flavonoids and carotenoids (Liu et al., 2004). The flower with methanol-HCl (0.1% HCl v/v) for 3 h. The extracts
stalk color of Chinese kale may change from green to mauve were measured at 530 and 600 nm using a spectrophotometer
when P is deficient. This color change seriously affects the (Thermo Spectronic Helios Ȗ, Madison, USA), and anthocyanin
visual quality of the flower stalks in the green-type Chinese concentration was calculated as A530-A600. After 50 ȝL
kale. extract was added to 4.5 mL methanol-HCl (0.1% HCl v/v),
In this study, the aforementioned 2 types of Chinese kale soluble phenols were measured at 280 nm. Flavonoid levels
were grown under hydroponic conditions, to study the effect were measured at 325 nm after 1 mL extract was added to 4
of P nutrition on plant growth and pigment formation in mL methanol-HCl (0.1% HCl v/v). The pH value was measured
flower stalk epidermis. The aim is to clarify how P nutrition after 2.0 g of flower stalk epidermis was homogenized with
regulates the synthesis of pigment, especially anthocyanins, 10 mL double-distilled water.
in flower stalk. Enzyme activity related to anthocyanin biosynthesis was
measured using the epidermis sample of plant flower stalk.
Materials and methods A total of 0.5 g sample was added to 5 mL of extract solution
consisted of 0.05 M Na2HPO4 (pH 7.0), 0.05 M ascorbic
3ODQW 0DWHULDO DQG 7UHDWPHQW acid, and 0.018 M mercaptoethanol, and then homogenized
The study was conducted in a plastic greenhouse located at 4°C, and centrifuged at 15000 rpm for 20 minutes. The
at the experimental vegetable farm of the South China supernatant was a crude enzyme extraction solution, which
Agricultural University. Two Chinese kale cultivars ‘Jianyexia’ was used to determine the activity of phenylalanine ammonia-
with green flower stalks and ‘Hongjiao’ with mauve flower lyase (PAL) and chalcone isomerase (CHI). PAL activity
stalks, were seeded in plastic plug trays with 72 cells. In was measured according to the method of McCallum and
total, 15 seedlings with 3 true leaves were transplanted in a Walker (1990), and CHI activity was determined according
plastic box (61 cm length × 42 cm width × 8 cm height) to the method of Boland and Wong (1975).
filled with 15 L nutrient solution. The nutrient solution
consisted of : N, 200; P, 30; K, 240; Ca, 130; Mg, 48; Fe, 2; 6WDWLVWLFDO $QDO\VLV
-1
B, 0.5; Mn, 0.5; Zn, 0.05; Cu, 0.02; and Mo, 0.02 mgL . All data were statistically analyzed by ANOVA, using the
The nutrient solution was aerated for 15 minutes every hour SPSS software package (version 16.0 for Windows). Two-way
via an automatically controlled pump, and was completely ANOVA was used to evaluate the effect of P levels and
refreshed every 9 days. cultivars, and their interactions were evaluated using
Three different P treatments were used: 30 (normal-P), Duncan’s multiple range tests.
-1
7.5 (low-P), and 0 mg·L (P-deficient). There were 4 replicates
of each treatment (one plastic box represented one replicate) Results
and for each cultivar, with a randomized block arrangement.
3ODQW *URZWK
6DPSOH &ROOHFWLRQ DQG $QDO\VLV Growth characteristics (e.g., plant height, stem diameter,
Chinese kale plants were sampled once the mature stage and leaf number) of the 2 Chinese kale cultivars were
was reached to measure their growth parameters. Three significantly affected by different P levels (Table 1). All
representative plants from each replicate were selected at indicators showed the same tendency in both cultivars: plant
random. ‘Jianyexia’, an early maturing cultivar, was sampled height, stem diameter and leaf number were not significantly
earlier than ‘Hongjiao’, a medium maturing cultivar. Plant different between normal P and low-P treatments, but
th th
height and stem diameter (in the middle of 5 and 6 nodes significantly lower and less in the P-deficient treatment than
of the flower stalk) were measured, respectively. Each plant other 2 treatments.
was divided into root, rootstock, and flower stalk respectively, P levels had a significant effect on growth of both Chinese
and fresh weight and dry weight (after drying at 70°C to kale cultivars. Shoot fresh weight gradually decreased with
constant weight) were obtained. the decreasing P levels, with a significant difference being
The epidermis of the flower stalk was used to measure the recorded among all 3 treatments. Compared with the normal-P
pigment content. Approximately 0.5 g fresh ground epidermis treatment, the shoot fresh weight of low-P and P-deficient
was extracted in 80% acetone, and the chlorophyll and treatments was reduced by 23.2% and 58.9% in ‘Jianyexia’,
carotenoid content were assayed according to Lichtenthaler respectively, and by 29.0% and 84.7% in ‘Hongjiao’, re-
(1987). The contents of soluble phenols, flavonoids, and spectively. A similar trend was observed for the effect of P
anthocyanins were analyzed according to Pirie and Mullins levels on root fresh weight.
Hort. Environ. Biotechnol. 54(3):243-248. 2013. 245
Table 2. The effect of P level on pigment content of the flower stalk epidermis in Chinese kale.
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'HILFLHQW 3 s E s E s D s D s D s D
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16
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The root/shoot ratio of both Chinese kale cultivars was had significant interaction effects by P levels and cultivars.
significantly higher in the P-deficient treatment than the However, yield, plant height, stem diameter, and leaf number
normal-P and low-P treatments; however, the difference had no interaction effect.
between the latter two was not significant. This result
indicated that P deficiency promotes limited biomass production 3LJPHQW RI )ORZHU 6WDON (SLGHUPLV
to be preferentially allocated to the roots. The root/shoot The chlorophyll and carotenoid contents in the flower
ratio of ‘Hongjiao’ was higher than ‘Jianyexia’ at the same stalk epidermis were not significantly affected by P levels
P level. (Table 2), with the exception that the P-deficient treatment
The yield (weight of flower stalks) gradually decreased significantly reduced the chlorophyll content in ‘Hongjiao’.
with decreasing P levels in both cultivars. Compared to the The ratio of chlorophyll a/b gradually decreased with decreasing
normal-P treatment, the yield with low-P and P-deficient P levels, and was significantly lower in the P-deficient
treatments was reduced by 22.0% and 65.4% in ‘Jianyexia’, treatment than the normal-P treatment. This result indicated
respectively, and by 18.8% and 80.4% in ‘Hongjiao’, that although there was no decrease in the total chlorophyll
respectively. There was a significant difference among the 3 content of the flower stalk epidermis, chlorophyll a content
treatments. was lower in the P-deficient treatment.
Two-way analyses (P levels and cultivars) were conducted The contents of soluble phenols, flavonoids, and antho-
for the growth parameters of Chinese kale, with the results cyanins in the flower stalk epidermis generally increased
showing that both factors significantly affect plant growth. with decreasing P levels. The pigment content was significantly
Shoot and root fresh weight, along with the root/shoot ratio, higher in the P-deficient treatment than the normal-P treatment.
246 Riyuan Chen, Shiwei Song, Xiuchun Li, Houcheng Liu, Danfeng Huang
Fig. 2. The effect of P level on the activity of 2 enzymes related to anthocyanin biosynthesis in Chinese kale. The vertical bars represent
the standard error of measurements (n = 4) and the values followed by different letter are significantly different at P < 0.05 (Duncan’s
method).
Hort. Environ. Biotechnol. 54(3):243-248. 2013. 247
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