BioFormosa（2012）47: 9-13
Newly Understanding of Kalanchoe pinnata, a CAM Plant
Fen-Wan Chen1, Meng-Yuan Huang2, Hsueh-Wen Yeh1, Chi-Ming Yang2*,
Yung-Ta Chang3*
1
Reseachrer of Institute of Earth Sciences, Academia Sinica
Taipei, Taiwan
2
Research Center for Biodiversity, Academia Sinica
Taipei, Taiwan
3
Department of Life Science, National Taiwan Normal University
Taipei, Taiwan
(Received: 8 January 2013, accepted: 27 March 2013)
ABSTRACT
Kalanchoe pinnata, a CAM plant was evaluated through the stable carbon isotopic analysis to
distinguish whether an obligate CAM plant or not. The δ13CPDB values of Kalanchoe pinnata are range
from –22.0 to –29.1 ‰ thus to be suggested to a facultative CAM plant. Also how environmental factors:
wind affect the δ13CPDB values of plants, was evaluated through experiments. The δ13CPDB values of
Kalanchoe pinnata are range from –23.9 to –29.1 ‰ in a wind-blowing treatment and –22.0 to –27.5 ‰
in the control.
Key words: Kalanchoe pinnata, obligate CAM plant, facultative CAM plant, carbon isotope, δ13CPDB
environmental changes
Introduction
For years photosynthesis has been discussed
and research wildly. Photosynthesis pathway can be
divided into 3 kinds of pathways: C3 pathway
(Calvin Cycle), C4 pathway, Crassulacean Acid
Metabolism (CAM) pathways. The metabolic
mechanism of CAM pathway in succulent plants
was wildly discussed from late 1950’s to early
1960’s (Bruinma, 1958; Ranson and Thomas, 1960;
Wolf, 1960; Walker, 1962). Furthermore, things are
more interesting since the improvements of
biochemical techniques in late 1980’s. On the other
hand, the techniques of stable isotopic analysis
were fully developments for over 20 years. The
differences in the carbon isotopic composition of
plants can be expressed as δ13CPDB values: δ13CPDB
(‰)= [(13C/12C(sample)-13C/12C(standard))/13C/12C(standard))]
×103. PDB stands for Pee Dee Belemnite. The
standard used to report δ13C-values as given above
is the 13C/12C ratio of limestone, composed of
Belemnite (Belemnite Americana) remains, from
the Pee Dee Formation, South Carolina (Craig,
1957). The techniques of carbon isotopic analysis
became a very useful tool to distinguish C3, C4, and
CAM plants. The δ13CPDB values of C3 plants range
from –22.0 to –38.0 ‰ (Yah and Wang, 2001), C4
plants from –10 to –20 ‰ (Bender, 1971), CAM
plants from –13 to –30 ‰. Due to the curiosity of
how plants represent the climate in ancient world,
and interested in how the carbon assimilation acts
and would environmental factors make changes in
CAM plant, experiments were done in a series of
process.
Materials and Methods
The Kalanchoe pinnata (Lam) Pers.
(Crassulaceae), is a succulent-leaved plant (Winter
et al., 1997), suggested to be whether an obligate
Crassulacean Acid Metabolism plant (Zotz
&Winter 1993) or a facultative CAM plant (Winter
et al., 1982). Some references (Kondo et al., 1998)
classify it as a PCK (Phosphoenolpyruvate
carboxykinase) CAM plant. Life plants were grown
from leaf cutting under our indoor environment.
They have become 3~4 leave plants after one
month, then transplanted to 3-inch pots. After 7
*Corresponding author: Yung-Ta Chang; FAX: 886-2-29312904; E-mail: [email protected]
Chi-Ming Yang; FAX: 886-2-23142234; E-mail: [email protected] Fen-Wan Chen, Meng-Yuan Huang, Hsueh-Wen Yeh, Chi-Ming Yang, Yung-Ta Chang days, experiment started with 18 days of time. Our
environment maintained in 22-23℃ photoperiod
10hr/14hr, light intensity about 3000 lux, and with
irrigation once a week. No fertilization was made
through the whole experiment. Tissue harvested at
different day in the control environmental was
dried for 48 h at 65°C, weighed, and ground to a
fine powder using a mortar and pestle.
Results
The experiment was lasted for 18 days, since
Feb. 25, 2000 to April 25, 2000. The sampling was
made out with the 11th, 13th, 18th days, and only the
mature leaves were harvested. During the
experiment, the average temperature was 22.5℃
(control 20.6 ℃ ; wind blowing 22.5 ℃ ), light
intensity were 3238 & 3164 lux, wind velocity
were 2.19 and 0.1 m/s (Table 1.). Irrigation was
made once a week. The estimate that Kalanchoe
pinnata is an obligate CAM plant or a facultative
CAM plant can be clearly understood through the
isotopic analysis. After one week putting in an 60℃
oven, the samples were powdered, and being
analysis by the stable carbon isotope analysis
system (Fig 1). The theorem of isotopic analysis is
samples are mixed with CuO in 1100℃, 10-2~ 10-3
conditions, processing redox reaction. After CO2
generate, collect and analyze them with MS: SIRA
system (Fig2). The δ13CPDB value of Kalanchoe
pinnata is varied from –23.9 to –29.1 in a contrast
and –22.0 to –27.5 in the control (Table 2).
Figure 1. Vacuum carbon isotopic analysis system in the
Institute of Earth Sciences, Academia Sinica.
Figure 2. MS (system SIRA) for carbon isotopic
analysis.
et al. (1982). (3) Environmental changes affect the
δ13CPDB values of Kalanchoe pinnata: In our
blowing experiment, the differences between
control and experimental are 2.0 to 2.3, suggesting
blowing in our nature has an effect on δ13CPDB
values of Kalanchoe pinnata. Since δ13CPDB values
reflect the contributions of the CO2 fixed in the
dark by PEP carboxylase and in the light by RuBP
carboxylase (O’Leary & Osmond, 1980), the
reason could due to wind effects the flowing of
CO2, and the velocity of CO2 speed up the formula
below toward right indirectly, but still needs to be
improved (Adams III & Osmond, 1988). Therefore,
more research will continue to go on in order to
clarify the environmental changes play what kinds
of role in the δ13CPDB values of Kalanchoe pinnata.
Thus we can understand the represent meaning of
plant’s fossils.
Discussion
There are some interesting findings: (1)
δ13CPDB values of Kalanchoe pinnata range from –
22.0 to –29.1; δ13CPDB values of Kalanchoe pinnata
in Bender et al. (1973) are –15.2 to –29.9. It seems
that δ13CPDB values of Kalanchoe pinnata have a
big difference among them. (2) According to the
definition of the obligate CAM plant, it would
process C4 pathway and have similar δ13CPDB values
with C4 plants – ranging from –10 to -20, but it
didn’t. The δ13CPDB values of Kalanchoe pinnata
fall in the range of C3 plant’s, it suggest that
Kalanchoe pinnata process Calvin cycle in our
experimental environment. According to the
definition of the facultative CAM plant, Kalanchoe
pinnata belongs to that. Similar suggest in Winter
Acknowledgements
10
Newly understanding of Kalanchoe pinnata Table 1. The control environmental conditions of absolute CAM plant (Kalanchoe pinnata)
Treatments
Indoor 7 days
Indoor 14 days
Indoor 21 days
Indoor 28 days
Woody box 7 days
Woody box 14 days
Woody box 21days
Woody box 28 days
Woody box-2 11 days
Woody box-2 13 days
Woody box-2 18 days
Wind velocity
Exp.(m/s)
Wind velocity
Control(m/s)
2.3
1.1
2.2
1.3
2.4
2.9
2.4
2.4
2.6
2.6
2.1
0.2
0.1
0.1
0.1
0.1
0.1
0.2
0.1
0.1
0.1
0.2
Average temperature
Exp.(℃)
21.6
21.7
22.7
23.8
22.0
22.6
23.7
23.3
23.0
23.4
22.4
Table 2. The δ13CPDB (‰) of absolute CAM plant
(Kalanchoe pinnata)
Treatments
Indoor 7 days
Indoor 14 days
Indoor 21 days
Indoor 28 days
Woody box 14 days
Woody box 21days
Woody box 28 days
Woody box-2 11 days
Woody box-2 13 days
Woody box-2 18 days
δ13CPDB Control (‰)
δ13CPDB Exp.(‰)
-27.5±0.1
-25.9±0.2(3)
-25.5
-23.4
-22.0±0.0(2)
-22.5±0.1(2)
-22.5±0.0(2)
-29.1
-28.3±0.0(2)
-27.3±0.1(2)
-25.7±0.2(3)
-26.4
-23.9
-24.0±0.1(2)
-24.8±0.0(2)
-24.7±0.1(2)
Average temperature
Control(℃)
20.7
21.7
22.8
23.6
22.0
21.2
23.7
23.3
23.0
23.4
22.4
Craig H. 1957. Isotopic standards for carbon and
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structure and immunogold localization of
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crassulacean acid metabolism of Kalanchoe
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Winter K, Foster JG, Schemitt MR and Edwards
We thank the technical support from Institute
of Earth Sciences, Academia Sinica. Also thanks to
the financial assistance from the National Council
of Science.
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11
Fen-Wan Chen, Meng-Yuan Huang, Hsueh-Wen Yeh, Chi-Ming Yang, Yung-Ta Chang light period: metabolic and stomatal control of
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12
生物學報（2012）47：9-13
有關 CAM 植物, 燈籠草( Kalachoe pinnata ) 的一些新了解
陳芬莞 1 黃盟元 2 葉學文 1 楊棋明 2* 張永達 3*
1
2
中央研究院地球科學研究所
中央研究院生物多樣性研究中心
3
國立臺灣師範大學生命科學系
（收稿日期：2012.7.25，接受日期：2013.3.15）
摘
要
CAM 植物燈籠草( Kalanchoe pinnata )經由穩定性碳同位素分析方法來分別其是否為絕對
CAM 植物或是非絕對 CAM 植物。實驗結果其 δ13CPDB 值介於-22.0 ‰與-29.1 ‰之間，因此判定為
非絕對性 CAM 植物。另外為了了解環境因子-風，對植物碳同位素值之影響，針對其實驗之結果
顯示，有風處理 δ13CPDB 值為-23.9 至-29.1 ‰，無風處理 δ13CPDB 值為-22.0 至-27.5 ‰。
關鍵詞：燈籠草、絕對 CAM 植物、非絕對 CAM 植物、穩定性碳同位素、環境改變、δ13CPDB
＊
通信作者：張永達（Yung-Ta Chang）；FAX：886-2-29312904；E-mail：[email protected]
楊棋明（Chi-Ming Yang）；FAX：886-2-29312904；E-mail：[email protected]