全 文 ：Dynamic Change of Four Physiological Indices of the
Penthorum chinense Pursh Yellow Seeding in the
Procedure of Green-turning
CHEN Guo-dong1，YANG Jun1，2* ，LIU Hui1，SHEN Shun1，QIN Feng1
1． College of Life Sciences，China West Normal University，Nanchong，Sichuan 637009，China;2． Institute of Rare Animal and Plant，West
Normal University，Nanchong，Sichuan 637009，China
Abstract ［Objective］The effect of the light on Penthorum chinense Pursh growth and development was researched． ［Method］The plantlet and
yellow seedling of Penthorum chinense Pursh were obtained by means of the technique of tissue culture． Once sampling for each 6-day was done，
respectively，with total of six-sampling． The dynamic change in the content of proline，chlorophyll，soluble protein and the activity of POD in the
stem of its yellow seedling during green-turning was tested． The content of soluble protein was measured with the method of Coomassie Brilliant
Blue G-250 Staining;the activity of POD，with Guaiacol Colorimetry;the content of free proline，with 3% Sulfosalicylic Acid;and the content of
chlorophyll，Ethanol Extraction． ［Results］The content of chlorophyll and soluble protein，and the activity of POD in the yellow seedling were lower
than those in normal seedling，while the content of proline was higher than that in normal seedling． In 30 days after the yellow seedling was placed
under lighting condition，the content of chlorophyll and the activity of POD was rising continuously，ultimately，similar to those in normal seedling;
the content of soluble protein and praline was firstly rising and then decreasing，ultimately near that in normal seedling，which indicated that the sys-
tem of photomorphogenesis in the yellow seedling was basically completed．
Key words Penthorum chinense Pursh;Yellow seedling;Physiological indicator;China
Received:December 14，2009 Accepted:January 3，2010
Foundation Project:the Project from the Sichuan Province for the
Construction of Subject: Research on the Protection and Utilization
of Wild Animal and Plant(SZD0420)．
* Corresponding author． E-mail:yangjun@tom． com
Penthorum chinense Pursh is the species belonging to
Saxifragaceae Penthorum，and also， is the traditional Miao
medicine，which whole plant is popularly used as medicine in
China． In the theory of traditional Chinese medicine，its wholly-
grass is with the character of warm，sweet，non-toxic，hot-re-
leasing，diuresis，detoxification，blood-activating，liver-balan-
cing， spleen-invigorating， jaundice-eliminating and other
effects［1 －2］．
Penthorum chinense Pursh always grows in a wild condi-
tion，which is mainly distributed in Northern，Eastern and
Southern Shaanxi，Sichuan and Guizhou and other provinces．
In recent years，its domesticated cultivation has been started in
Sichuan Province，meanwhile，the research on its multiplication
by means of the technique of tissue culture and its physiologi-
cal /biochemical analysis were carried out． The plant has the
strong demand for light during its growth and development． The
growth and development of the plant are regulated by the
change of light quality，light intensity and irradiance［3］． There is
a relevant long-term process from its yellow seedling being
turned into the normal green plant with the ability-building in
photosynthesis，including the synthesis of chlorophyll，the for-
mation of photosynthetic apparatus，the development of true
leaf and so on． Therefore，the differentiation，development，
material transportation and many other physiological phenome-
na are involved in the green-turning process［4］． The Penthorum
chinense Pursh being taken as the experimental material，the
dynamic change in the content of soluble protein，chlorophyll，
proline and the activity of POD in the stem of its yellow seedling
during the process of green-turning was studied in order to ex-
plore the impact of light on its growth and development．
1 Materials and methods
1． 1 The experimental material The axillary buds，which
were cut from the sterile Penthorum chinense Pursh seedlings
that were normally growing in the room with temperature of
25 ±1 ℃ and light of 4 000 lx，were cultured in the hormone-
free MS medium in culturing bottle． 50 culturing bottles were
placed under the lighting condition of 4 000 lx and another 50
culturing bottles，under the condition of darkness． 50 culturing
bottles under the dark condition，inside which，the height of
yellow seedlings was about 7 cm after about 30 days culturing，
were moved under the lighting condition of 4 000 lx and continu-
ally cultured．
The section of all plants in 100 culturing bottles that both
were placed under dark firstly and then lighting condition，and
had been put under lighting condition were respectively sampled
on the 0，6 th，12th，18th，24th and 30th day after the yellow
seedling was started to be placed in the lighting cultivation and
there were 3 repetitions for each sample．
1． 2 The Testing method for proline，chlorophyll，soluble
protein and POD The content of soluble protein was deter-
mined with the method of Coomassie Brilliant Blue G-250 Stai-
ning［5］;the activity of POD，with Guaiacol Colorimetry［6］;the
Responsible editor:Xia Jing Responsible translator:Huang ZhongxiangMedicinal Plant /药用植物研究 2010，1(1):6 －8，23
content of free proline，with 3% Sulfosalicylic Acid;the content
of chlorophyll，with Ethanol Extraction［7］．
1． 3 The data treatment The content of physiological indica-
tors and the activity of POD in different time periods could be
calculated after all of the tested values were put into the formu-
la，respectively．
2 Results and analysis
The Figures of the dynamic variation of four physiological
indices were made according to the corresponding values in the
Table 1．
Table 1 The Effect of the Different Light Durations on the Physio-
logical Index of the Penthorum chinense Pursh Yellow
Seeding in the Procedure of Green-turning
Sam-
plinge
Light
duration
d
POD
activity
U/g
Proline
content
μg/g
Soluble
protein content
mg/g
Chloroplast
content
mg/g
Normal 0 475． 08 32． 21 3． 07 1． 69
Growth 6 425． 48 46． 06 3． 49 1． 71
12 387． 27 84． 18 3． 52 1． 73
18 523． 40 62． 35 4． 02 1． 88
24 543． 28 52． 40 3． 65 1． 93
30 691． 35 65． 36 4． 39 2． 09
Yellow 0 96． 20 128． 45 1． 18 0． 48
Seeding 6 123． 76 134． 78 2． 09 0． 52
Growth 12 284． 14 111． 47 2． 33 0． 75
18 324． 34 254． 50 2． 89 1． 03
24 734． 88 122． 85 7． 35 1． 55
30 810． 76 55． 69 4． 88 2． 13
As can be seen from the Fig． 1，the activity of POD in the
yellow seedlings was lower;the activity of POD was gently in-
creasing within the first day to 18th day after the yellow seed-
lings were moved under lighting condition;the activity of POD
was rising rapidly within 18th －24th day，within 24th －30th day，
the activity-increasing of POD was slowing down and eventual-
ly its value was same as that of the CK and even slightly high-
er than that of the CK．
Fig． 1 The dynamic change of the activity of POD during
the yellow seedlings green-turning
From the Fig． 2，it can be seen that the content of proline
in the yellow seedling was higher than that in CK． The content
of praline in the yellow seedling had always been higher than
the normal value within 0 － 24th day after lighting cultivation，
which was close to that in CK until the 30th day． Within 0 －12th
day，the content of proline was little rising at first and then
slightly decreasing． There was a sharp increment in its content
from the 12th day，then，a significant decline appeared from
the 18th day，and finally，the content of proline was close to
that in CK until the 30th day．
Fig． 2 The dynamic change of the content of proline dur-
ing the yellow seedlings green-turning
Fig． 3 The dynamic change of the content of protein dur-
ing the yellow seedlings green-turning
Fig． 4 The dynamic change of the content of chloroplast
during the yellow seedlings green-turning
The content of chlorophyll in the yellow seedling was lower
than that in CK． The content of chlorophyll had been in a clear
upward trend but lower than that of normal value before the
18th day from the lighting cultivation． There was a sharp incre-
ment in its content within 18th －24th day;decline from the 24th
day and the content of chlorophyll was close to that in CK until
the 30th day．
3 Conclusions and discussions
It was considered by Dr． Ridge I． Q． etc． that the imbal-
7CHEN Guo-dong et al． Dynamic Change of Four Physiological Indices of the Penthorum chinense Pursh Yellow Seeding in the Procedure of Green-turning
ance between the formation and removal of reactive oxygen in-
side plants resulted from the adverse environmental condition，
which would damage the plant due to the accumulation of free
radical［8］． The POD，an important antioxidant enzyme，can
effectively remove H2O2 and O
2 － etc in plant so that the mem-
brane lipid peroxidation would be reduced for the protection of
photosynthetic apparatus from being harmed． The capacity in
the elimination of the reactive oxygen has greatly been weak-
ened because the system of the active oxygen metabolism in-
side the body of yellow seedling is out of balance．
It was found in the experiment that the light-prohibiting
could be formed to the yellow seedling that was moved into the
normal lighting condition after its long-term treatment under the
condition of darkness and the extra light would produce the re-
active oxygen in species through the approach of Mehler Re-
action［9］ ． It was possible for the plant to be prevented from the
photo-oxidation damage through the approach that the activity
of POD in the yellow seedling that was exposed under light
was significantly increased，which was an adaptation strategy
of plant protection［10］． However，the activity of POD in the
yellow seedling would also gradually be stepped into normal level
with the gradual adaptation of the plant to light environment．
The free proline，an important osmotic and adjusting sub-
stance in plant cell，is closely related to the elimination of the
free radical of active oxygen and the reduction of membrane lip-
id peroxidation in plant body［11］． Generally，the content of pro-
line in plant is very low，but it would dramatically be increased
under the adversity． The high contain of proline in the yellow
seedling in the experiment would be beneficial to its tolerance or
adaptability-strengthening to adversity． The further accumula-
tion of the proline in yellow seedling may be owing to what it
was difficulty for the yellow seedling to adapt this sudden
change resulted from yellow seedling being moved from the
dark to the light;and eventually，the content of the proline was
decreasing to the normal level in plant after the yellow seedling
was adaptable to the normal light in its external environment．
Most of the soluble proteins in plant are the enzymes in-
volved in various metabolisms． And its content can reflect the
metabolic situation of plant［12］． Under dark environment，the
chloroplast grows abnormally and the activity of many kinds of
enzymes is very low． The various metabolisms in the yellow
seedling were strengthened with the gradual formation of photo-
morphogenesis of the plants moved from the dark condition to
the lighting condition． However，with the complete adaptation of
the plant to external environmental condition，the various meta-
bolic activities would ultimately be normalized［13］． It is a specific
example of metabolic process above-mentioned that there was
a trend that the content of soluble protein in the yellow seedling
of Penthorum chinense Pursh，after which was placed under
the lighting condition from the dark，was increasing at first，
then decreasing and finally toward same variation as the CK．
The biological synthesis of organic matter in plant is affect-
ed by the chlorophyll and also the content of chlorophyll is an
important issue in the process of the green-turning of yellow
seedling． The light is the main condition affecting the synthesis
of chlorophyll and the light can make the origin of dephytolchlo-
rophyll reverted into the dephytolchlorophyll［14］． The function of
chlorophyll in the growing plant under dark condition is inhibi-
ted，resulting in what the synthesis of chlorophyll can almost
not be conducted． The ability in the production of chlorophyll
was rapidly be recovered because the chloroplast was activated
and proliferated as soon as the yellow seedling was moved un-
der lighting condition，therefore，the content of chlorophyll was
rising and reached to the normal level in about 30th day after it
was moved in light．
The results showed that in the process of the green-turning
of Penthorum chinense Pursh yellow seedling，the above-men-
tioned four physiological indexes were beneficial to the direction
of photosynthesis，antioxidant and detoxification，and ultimate-
ly，were close to the corresponding indicators of green seedling
under normal lighting condition． The conclusion was same as
what Yang and others had done in the study on the differentially
expressing proteins in the process of the yellow-removing of
rice seedling that the yellowing process was not only closely re-
lated with the mechanism of photosynthesis［15］，but also the re-
acting status gradually being relieved of the oxidative stress of
the yellow seedling in the period．
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(To page 23)
8 Medicinal Plant 2010
Fig． 2 Heavy metal cluster analysis of Periploca forrestii
Schltr．
for heavy metal detecting and controlling of the TCM． There-
fore，for producing a high-quality TCM，we should respect the
TCM development history and detect the soil and water condi-
tions at the same time in selecting standardized planting base
which should have fresh air，pure water，unpolluted soil and
good agro-ecological environment． By implementing GAP for
Chinese Crude Drugs and its accreditation to control the heavy
metal contents strictly，we should，simultaneously，found a
green TCM production base with rational use of pesticides and
fertilizers and exert to avoid to be polluted in harvesting，trans-
porting and processing．
References
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herbal medicine［J］． Trends in Pharmacological Sciences，2002，23
(3):136．
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control of contamination of Chinese medicinal plants by heavy met-
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nese Herbal Drugs and Ethnic Medicinal Materials［S］． Guiyang:
Guizhou Technology Publisher，2003:151． (in Chinese )．
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(in Chinese )
檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪
．
黑骨藤药材中铅·镉·砷·汞·铜的含量及其质量标准评价
陈华国1，2，曹桂红1，2，赵 超1，2，周 欣1，2*
(1．贵州师范大学天然药物质量控制研究中心，贵州贵阳 550001;2．贵州省山地环境信息系统与生态环境保护重点实验室，贵州贵阳 550001)
摘要 ［目的］对不同产地的黑骨藤药材中铅、镉、砷、汞、铜的含量进行测定，为高品质黑骨藤药材的筛选、药材质量标准建立及中药材生产质
量管理规范(GAP)种植提供科学依据。［方法］样品经湿法消解后，用原子荧光光度法和原子吸收分光光度法进行检测;采用聚类分析等数量
统计软件对测定结果进行分析处理。［结果］黑骨藤药材中主要是铅和镉重金属元素超标，超标率为铅 47． 62%、镉 71． 43%、汞14． 28%、铜
14． 28%。［结论］不同产地的黑骨藤药材中砷、汞和铜含量较低，铅和镉含量严重超标。
关键词 黑骨藤;原子荧光光度法;原子吸收分光光度法;重金属;
欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁
含量测定
(From page 8)
扯根菜黄化苗转绿过程中 4 种生理指标的动态变化
陈国栋1，杨 军1，2* ，刘 慧1，沈 顺1，秦 凤1
(1．西华师范大学生命科学学院，四川南充 637009;2．西华师范大学珍稀动植物研究所，四川南充 637009)
摘要 ［目的］探讨光照对扯根菜生长发育的影响。［方法］采用组织培养法获取扯根菜组培苗和黄化苗。每隔 6天分别取样，共 6 次。研究扯
根菜黄化苗转绿过程中茎段内脯氨酸、叶绿素、可溶性蛋白含量以及 POD活性的动态变化。可溶性蛋白含量测定采用考马斯亮蓝 G-250 染色
法;POD活性测定采用愈创木酚比色法;游离脯氨酸含量测定采用 3%磺基水杨酸法;叶绿素测定采用乙醇提取法。［结果］黄化苗的叶绿素和
可溶性蛋白含量以及 POD活性均低于正常苗，而脯氨酸含量高于正常苗;黄化苗见光后的 30 d内，叶绿素含量和 POD活性一直处于上升趋势，
并最终与正常苗相近;可溶性蛋白与脯氨酸的含量总体呈现先升后降的趋势，最终也接近正常苗;表明黄化苗的光形态建成基本完成。
关键词 扯根菜;黄化苗;生理指标
．
32CHEN Hua-guo et al． Determination and Quality Standard Evaluation of Pb，Cd，As，Hg and Cu in Periploca forrestii Schltr．