全 文 ：Study on the Flower Bud Differentiation Process
of Gentiana triflora Pall.
Yan SUN1*, Н.А. Царенко2
1. Heilongjiang University, Harbin 150080, China;
2. Far Eastern National University, Vladivostok 690950, Russia
Supported by the Youth Fund of Heilongjiang University, China.
*Corresponding author. E-mail: sy81518@sohu.com
Received: December 12, 2011 Accepted: December 25, 2011A
Abstract [Objective] This study was to obseve the flower bud differentiation process
of Gentiana triflora Pall., and to clarify the flower bud development law and the cor-
relation between flower buds differentiation and external morphology. [Method] The
paraffin section method was used to observe the flower bud of Gentiana triflora un-
der microscope. [Result] The flower bud differentiation process of Gentiana triflora
could be divided into six stages: the undifferentiated stage, initial differentiation stage,
sepal primordia differentiation stage, petal primordia differentiation stage, stamen pri-
mordia differentiation stage and the pistil primordia differentiation stage. The flower
bud differentiation started from early May, when the plant had 10-11 stem nodes,
and completed in early June, lasted about 40 d. The effect of plant height on flower
bud differentiation was not significant. [Conclusion] The study provided theoretical
basis for the high-yield cultivation, variety selection and flowering regulation of Gen-
tiana triflora.
Key words Gentiana triflora; Flower bud; Development
Agricultural Science & Technology, 2012, 13(3): 558-560, 564
Copyright訫 2012, Information Institute of HAAS. All rights reserved Agronomy and Forestry
G entiana triflora Pall. is aherbaceous perennial plant inthe genus Genitiana, mainly
distributed in Inner Mongolia, Hei-
longjiang, Liaoning, Jilin, Hebei of Chi-
na. It is also found in Japan, Korea,
Russia [1]. The dried root and rhizome
can be used as medicine, which are
the main composition of Guanlong-
dan, the native northeastern medici-
nal materials. With long medicinal his-
tory, Guanlongdan is effective to
purge liver-fire, abate deficiency-heat.
The main constituent of Gentiana triflo-
ra is gentiopicrin, which is hepatopro-
tective, cholagogic, diuretic, and anti-
hypertensive, as well as effective on
anti-inflammatory and antibacterial[2-3].
Currently, studies on Gentiana
triflora mainly focus on its pharmaco-
logical efficacy, chemical composition
and cultivation, while relative few have
devoted on flower development and
reproductive biology[4-8], and there is no
report on flower bud differentiation.
This study conducted preliminary re-
search on the flower bud differentiation
process of Gentiana triflora, with the
aim to clarify the flower bud develop-
ment law and the correlation between
flower buds differentiation and external
morphology, in order to provide theo-
retical basis for the high-yield cultiva-
tion, variety selection and flowering
regulation of Gentiana triflora.
Materials and Methods
Material
The experimental material was
the perennial wild Gentiana triflora
plant grown at the Vladivostok suburb
of Primorsky Krai, Russia.
Methods
The experimental materials were
taken every 3-5 d from the late April
2010 to the end of June, and ten main
stem buds were randomly taken each
time to fix by the FAA fixative; the
number of stem nodes and height of
the plants were measured while taking
the experimental materials. Stained by
the hematoxylin, the buds were cut in-
to 10 -12 μm thick slices using con-
ventional paraffin sectioning, and then
mounted the slices with neutral gum.
The middle parts of slices in continu-
ous wax band were taken to observe
and photograph under the LEICA-DM
LB2 digital microscope.
Results and Analysis
Undifferentiated stage
In early April, with the germination
of overwintering bud, the stem height
gradually increased, and the cell activ-
ity of meristem in terminal bud became
active. On the longitudinal section, the
growth cone of stem terminal bud was
presented oblateness shape, with
longer width than height, and the cells
in the meristem were small, similar in
shape, arranged in neat rows, with
larger nucleus and dense cytoplasm
(Fig.1-1). There were many leaves
concentrated at the top of the stem
during this period. And with the growth
of growth cone, the leaf primordia con-
tinued to differentiate.
Initial differentiation stage
The growing point protruded at
the top of the stem became broad and
large, upheaved upward, presenting
nearly hemispheric shape (Fig.1-2),
later, it gradually elongated, widened,
and became flat (Fig.1-3, 4), marking
the beginning of the transformation
from vegetative growth to reproductive
growth of the plant. This stage first ap-
peared on May 2, and climaxed at
around May 10. The leaves (including
scale leaves) at this period were 10-
11 pairs, and the plants were 12 -28
cm in height. This stage had much
longer duration which could extend to
May 25.
Sepal primordia differentiation
stage
The top of the elongated growing
DOI:10.16175/j.cnki.1009-4229.2012.03.031
Agricultural Science & Technology
Vol.13, No.3, 2012 Agricultural Science & Technology
2012
1, Undifferentiated stage; 2-4, Initial differentiation stage; 5, Sepal primordia differentiation stage; 6-7, Membrana intracalycina and petal
primordia differentiation stage; 8, Stamen primordia differentiation stage; 9, Pistil primordia differentiation stage; 10-11, The differentiated
two-spilt stigma; 12, Ovary wall and ovule.
Fig.1 Flower bud differentiation of Gentiana triflora Pall.
point became wide and flat, thereby
generating protuberance (Fig.1-4, 5
were the protruding sections on both
sides), which was the sepal primordia,
marking the beginning of sepal differ-
entiation stage. With the growth of
sepal primordia, protruding structure in
horizontal V shape, the membrana
intracalycina of Gentiana triflora, was
formed inside the sepal (Fig.1-6, 7).
This stage first appeared on May 15.
The leaves were 12-14 pairs, and the
plants were 18-39 cm high. This stage
extended to June 1.
Petal primordia differentiation stage
With the differentiation and con-
tinued growth of sepal primordia, the
protuberance generated inside the
stem base grew into the petal primor-
dia (Fig.1-6, 7). The continuous elon-
gation of petal primordia eventually de-
veloped into tubular bell-shaped corolla.
This stage mainly concentrated from
May 25 to June 4.
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Agricultural Science & Technology Vol.13, No.3, 2012
2012
Stamen primordia differentiation
stage
During the continuous growth and
development of petal primordia, new
protuberance generated inside the
continuously elongated petal primor-
dia, which was the stamen primordia
(Fig.1-8). Stamen primordia continued
elongating (Fig.1-9, 10), and finally
developed into five stamens distinct.
This stage was from June 2 to June 20.
Pistil primordia differentiation stage
Pistil differentiation stage mainly
concentrated from June 10 to June 29.
Inside the stamen primordia, upward
protuberance appeared in the central
part of the top receptacle, which was
the pistil primordia (Fig.1-8, 9). The
development on both sides of the pistil
primordia was faster, resulting in the
top depression of the growing point
(Fig.1-10), which further developed in-
to two split stigma and locule (Fig.1-
11). The size of the flower bud continu-
ously increased from early July. Sta-
men and pistil were still developing,
and pollen mother cell appeared in the
anther cell. The ovule in the ovary be-
gan to develop (Fig.1-12).
Conclusions and Discus-
sion
Flower bud differentiation and
flowering is a complex morphogenesis
process. It is the coordinated result of
various factors inside the flower which
are subject to the influence of external
environment, and the factors form a
complex network system to regulate
flowering[9]. Research on the differenti-
ation of Gentiana triflora is of important
significance in production, which is not
only the initial value to understand the
biological characteristics of the species,
but also can be used as the basis for
the development of Gentian cultivation
practices. Gentiana triflora, the root of
which can be used as medicine, is of-
ten cultivated with topping and inflo-
rescence removal in production to in-
hibit the reproductive growth and re-
duce nutrient consumption of the plant,
promoting root growth and increasing
root yieldandquality[10-12]. Regulating the
vegetative and reproductive growth of
Gentiana triflora based on the flower
bud differentiation characteristics to
promote the root growth is able to in-
crease the root production.
It is usually believed that the sign
of the beginning of plant flower bud dif-
ferentiation is the increase of bud size
and changes in top shape [13], which is
consistent with the result of this study.
As seen from the observation results,
the flower bud differentiation process
of Gentiana triflora could be divided in-
to six stages: the undifferentiated stage,
initial differentiation stage, sepal pri-
mordia differentiation stage, petal pri-
mordia differentiation stage, stamen
primordia differentiation stage and the
pistil primordia differentiation stage.
Floral bud begin to differentiate at the
beginning of May, and begin the pistil
primordia differentiation stage in early
June. It takes at least 40 d from the
initial differentiation stage to the
completion of the flower structure
differentiation.
Although the plant morphogene-
sis law depends on their hereditary,
the flower bud differentiation is also
subject to many inside and outside
factors, such as temperature, light,
nutrients and water[13-15]. The flower bud
differentiation of Gentiana triflora starts
from the beginning of May, lasting
about 20 d, and the sepal primordia
differentiation stage also lasts for near-
ly two weeks, which may be related to
local climatic conditions. The study re-
gion belongs to the temperate mar-
itime climate with earlier spring turning
green but relatively low temperature,
and low day-night temperature differ-
ence, resulting in slowed flower bud
growth and longer differentiation time.
Thereafter, the developing rate of petal
primordia, stamen primordial and pistil
primordia gradually speeds up with the
increase of temperature.
Generally, plant with less than
10-11 main stem nodes is at the veg-
etative stage, and then is the repro-
ductive stage, i.e., flower bud differen-
tiation stage. The effect of plant height
on flower bud differentiation is not sig-
nificant, therefore, using the number of
nodes as an indicator can determine
whether the plant enters the flower bud
differentiation stage.
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三花龙胆花芽分化过程的研究
孙阎 1*，Н.А. Царенко2 （1.黑龙江大学，黑龙江哈尔滨 150080；2.远东国立大学，俄罗斯符拉迪沃斯托克 690950）
摘 要 [目的]研究三花龙胆花芽分化过程，阐明其花芽发育规律及花芽分化与外部形态的相关关系。[方法]利用石蜡切片法对三花龙胆花芽进
行显微观察。[结果]三花龙胆花芽分化过程可分为 6个时期，即未分化期、分化初期、萼片原基分化期、花瓣原基分化期、雄蕊原基分化期、雌蕊原
基分化期。花芽分化从 5月初开始至 6月上旬基本完成，历时约 40 d。其植株发育到 10～11节时，花芽开始分化，花芽分化与植高关系不大。[结
论]该研究为制定三花龙胆丰产栽培措施、品种培育及花期调控等提供了理论依据。
关键词 三花龙胆；花芽分化；发育
基金项目 黑龙江大学青年基金项目。
作者简介 孙阎（1981-），男，黑龙江庆安人，副教授，从事植物发育生物学研究，E-mail: sy81518@sohu.com。*通讯作者。
收稿日期 2011-12-12 修回日期 2011-12-25
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Responsible editor: Na LI Responsible proofreader: Xiaoyan WU
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miltiorrhiza bunge root by raising and
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高斯序贯模拟与克里格插值 2种土壤重金属污染分析方法的比较
王倩，丁宁，孙英君 * （山东建筑大学土木学院，山东济南 250014）
摘 要 [目的]探讨克里格插值和高斯序贯模拟 2种方法在土壤重金属污染分析中的差异，以期为土壤重金属污染分析方法的选择提供参考依
据。[方法]以山东聊城某县重金属铜采样数据为研究对象，利用克里格插值和高斯序贯模拟 2种方法进行土壤重金属铜的空间分布建模模拟。并
选择 30个采样点作为交叉验证数据集，对 2种插值方法进行对比分析。[结果]克里格方法和高斯序贯模拟在均值段和极值段的模拟各有优势，研
究者应根据试验数据的特点及应用目的对二者进行选择使用。[结论]土壤重金属污染分析是土壤治理、生态修复的前提，该研究结果对于根据不
同目的需求来选择不同的插值、模拟方法具有重要意义。
关键词 土壤；重金属；高斯序贯模拟；克里格插值
基金项目 山东省科学技术发展计划（软科学部分）项目（2009RKB220）资助。
作者简介 王倩（1969-），女，辽宁大连人，讲师，硕士，从事城市信息系统分析及应用方面研究。*通讯作者，E-mail: sdjzusyj@126.com。
收稿日期 2011-12-12 修回日期 2011-12-27
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Responsible editor: Na LI Responsible proofreader: Xiaoyan WU
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