全 文 ： 第 26卷第 5期
2002 年 9 月
南 京 林 业 大 学 学 报(自 然 科 学 版)
Journal of Nanjing Fo restry University(Natural Sciences Edition)
Vol.26 , No.5
Sept., 2002
A Study on Variations of Physiological Indices of Cold
Resistance in Fukiencypress
HE Kai-yue1 , LI Xiao-chu2 ,HUANG Li-bin2 ,YANG Zong-wu3 ,YAN Hong-zhen1
(1.Nanjing Fo restry University , Nanjing 210037 , China;2.Jiangsu Academy of Fo restry ,
Nanjing 211153 , China;3.Fujian Academy of Forestry , Fuzhou 350012 , China)
Abstract:The results of the low-temperature(-8 ℃, -15 ℃)t reatment of the 1 ～ 2 a seedlings
of Fukiencypress demonstrated that , when treated for 24 hours , the contents of chlo rophyll and
proline in plants increased in comparison with the control of natural low temperature at -4 ℃
w ith the root vitality decreasing.Moreover , the dif ferences among provenance w ere quite marked.
The increase ratios in chlorophyll and proline were that the provenance of Xianyou and Nanjing
w ere higher than that of Dehua while the decrease ratios in root vitali ty were Xianyou
proline all increased during 24 hours;w ith markedly dropping in chlorophyll during 48 and 72
hours and markedly increasing in proline during 72 hours.The content of soluble sugar increased
af ter being t reated 48 hours and 72 hours , the content of MDA all increased during the f reezing
period.The synchronous observations made in the fields showed that , under the conditions of natu-
ral low temperature of -4 ℃ fo r 24 hours , only slightly f reezing w as observed in phyllades at the
top of some branches in 2 a Fukiencypress seedlings under -8 ℃ for 48 hours , and the freeze in-
jury occurred(freezing of the tips w as not observed fo r majori ty of the seedlings);while the g reat
majori ty of seedlings w ere f rozen to death under -8 ℃ for 72 hours and -15 ℃ fo r 48 and 72
hours.Variat ions in chlorophyll , proline and root vitality under low-temperature t reatment w ere
the regulating reactions of physiological mechanism of plants to the adverse atmospheric tempera-
tures.By use of the above-ment ioned variations of phy siolog ical indices under the low-temperature
stressed t reatment , i t is possible to seek the means for appraising the f reeze-resisting provenance
and to provide the scientific basis fo r int roducing the species northwo rds.
Key words:Fukiencypress;Freeze injury;Physiological indices of cold resistance
CLC number:S718 Document Code:A Article ID:1000-2006(2002)05-0010-05
福建柏抗寒生理指标变化研究
何开跃1 ,李晓储2 ,黄利斌2 ,杨宗武3 ,严洪真1
(1.南京林业大学 , 江苏 南京 210037;2.江苏省林业科学研究院 ,江苏 南京 211153;
3.福建省林业科学研究院 ,福建 福州 350012)
摘 要:对福建柏1 ～ 2年生苗木进行-8 ℃, -15 ℃低温胁迫处理试验测定表明 ,在处理24 h情况下 ,植株
体内叶绿素和脯氨酸的含量比对照(自然低温-4 ℃)都有所升高 ,而根活力指标均有所降低;且种源间差
异明显:叶绿素和脯氨酸含量的增加率为仙游和南靖种源均大于德化种源;根活力指标的降低率为仙游<
南靖<德化 。对仙游种源2年生苗木低温处理试验测定表明 ,叶绿素和脯氨酸含量在处理 24 h时均升高 ,
但在处理 48 h 、72 h时叶绿素含量明显下降;脯氨酸含量在处理 72 h时明显升高。可溶性糖含量在处理
Receive date:2001-09-21 Revised date:2002-04-16
Foundation Item:国家“九五”科技攻关项目“福建柏珍贵建筑材料树种良种选育及培育技术”内容之一
Biography:何开跃(1959-),女 ,四川人 ,南京林业大学森林资源与环境学院副教授,博士 ,主要从事生物化学和植物生理学的研究。
48 h和 72 h后有所升高。MDA含量在冷冻处理时均升高 ,田间同步观测表明 ,在-7 ℃24 h自然低温条
件下 ,2年生福建柏苗仅出现少量侧枝顶端鳞叶轻微受冻;-8 ℃持续 48 h自然低温 ,则出现中度冻害(多
数苗木未见顶梢受冻);而经-8 ℃处理72 h , -15 ℃处理48 h和 72 h的苗木绝大多数冻死 。低温处理福
建柏叶绿素 、脯氨酸和根活力指标的变化是植株适应不利气温的生理机制调节反应。利用低温胁迫处理
后上述生理指标的变化 ,可以寻求评估耐寒种源的途径 ,并为北移引种提供科学实验依据。
关键词:福建柏;冻害;抗寒生理指标
Fukiencypress(Fokienia hodginsii Henry Dunn et Thomas)is unique needle-leave t ree provenance of
the cypress family(Cupressaceae)in China , and is the rare t ree provenance protected in the first botch of the
second grade by state.Fukiencypress is naturally dist ributed in the hilly areas located in the northern part of
south subtropics and southern part of middle subtropics , respectively.They belong to the neutral , shallow-ro-
oted and slightly sun provenance;and the side roots are well developed w ith st rong penet rating power and
have no st rict demand on the soil and si te in forest land.It s adaptability and resistance to w ind and disease
are relat ively st rong
[ 1 ～ 3] .Therefore , under the present circumstances of advocating fo r af forestation w ith
multiple tree provenance , st reng thening the const ruct ion of the biolog ical environment and promoting the
sustainable development of forest ry , Fukiencypress has a broad prospect of application in hilly areas in the
southern part of our country owing to i ts high quali ty as timber and excellent provenance as landscape engi-
neering.For the purpose of introducing and enlarging cultivation of this rare , excellent tree provenance
northward ,we carried out the st ressed low-temperature t reatment of the 1 ～ 2 a seedlings so as to determine
variations of physiological indices and observed freeze injuries in the fields.The aim of the study on relation-
ship betw een variations of physiological indices above-mentioned and freeze injury is to provide the scientif ic
basis fo r screening freeze-resisting varieties and int roducing cultivars northw ard.
1 Material and Method
1.1 Natural conditions of test site for cultivation
The test site fo r seedling s is situated in the nursery of Jiangsu Academy of Fo restry located in 118°20′
E , 32°08′N.It is w ithin the climate of Northern subtropics w ith annual average temperature of 15.4 ℃, ex-
tremely low est and highest temperature of -14 ℃(approx imately -8 ℃ fo r normal years)and 40.7 ℃
(approximately 38 ℃ for normal years)respectively , annual precipitation of 1 026 mm and relative humidity
of 79%[ 4].
1.2 Test material and method
The seeds w ere supplied by Fujian Academy of Forest ry.They w ere sow ed in the nursery of Jiang su
Academy of Fo restry.The provenances are 1 a Xianyou , Nanjing , Dehua and 2 a Xianyou.The low-tempera-
ture t reatments were conducted in tw o batches:a)The low-temperature treatment of -8 ℃ and -15 ℃
was conducted wi th 1 a seedlings belonging to Xianyou ,Dehua and Nanjing and 2 a seedling to Xianyou for
24 hours(in winter , 1998);b)The same treatment of -8 ℃and -15 ℃was conducted with 2 a seedlings from
Xianyou for 24 ,48 and 72 hours(in winter ,1999).The control experiments were carried out under the natural low
temperature of -4 ℃.
Determination in chlorophyll , proline , sloluble sugar and MDA was made w ith reference to the method
established by Zou Qi
[ 5] .
Determination in root vi tality w as made w ith the method by Zhang Zhiliang
[ 6] .
2 Experimental Results
2.1 Variations in chlorophyll ,proline and root vitality under low-temperature treatment
The resul t show ed that , af ter being treated at -8 ℃ and -15 ℃ for 24 hours , contents of chlorophyll
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2002年 总第 101期 HE Kai-yue et al:A study on variations of physiological indices of cold resistance in Fukiencypress
in Fukiencypress all exhibi t a tendency of increase in comparison w ith the controlled g roup.The increase ra-
tio in chlorophyll is:-8 ℃>-15 ℃>-4 ℃(the natural low temperature as controlled);and the differ-
ence among provenances at -8 ℃ is Nanjing >Xianyou>Dehua;2 a seedlings>1 a seedlings , and at
-15 ℃,Xianyou>Nanjing>Dehua;and 1 a seedling s>2 a seedlings.
Contents of proline in Fukiencypress increased bo th in comparison wi th the natural low temperature.
The degree of increase w as -15 ℃>-8 ℃.In t reatment of both -8 ℃ and -15 ℃, the increase ratio in
content of proline was 2 a Xianyou>Nanjing>1 a Xianyou.
Roo t vitality of the seedlings of Fukiencypress decreased in comparison with the natural low tempera-
ture.The degree of decreas was -15 ℃>-8 ℃.According to both t reatment , the decrease ratio w as De-
hua>Nanjing>1 a Xianyou>2 a Xianyou.(refer to Table 1).
Table 1 Variations in chlorophyll , proline and root vitality under the low-temperature treatment of Fukiencypress(FW)
Provenance
Contents of Chlorophyll μg·g-1
CK -8 ℃ -15 ℃
Contents of proline μg·g -1
CK -8 ℃ -15 ℃
Root vitality μg·g-1
CK -8 ℃ -15 ℃
Dehua , 1a 0.67±0.01(4) 1.03±0.08(5)
53.7＊＊
0.88±0.12(5)
31.3 8.9±0.1(4)
10.1±0.1(5)
13.5＊＊
11.8±0.5(5)
32.6＊＊ 170±2(4)
130±3(5)
23.5＊＊
107±2(5)
37.1＊＊
Nanjing , 1a 0.58±0.04(4) 1.46±0.08(5)
151.7＊＊
0.95±0.09(5)
63.8＊＊ 7.3±0.3(4)
9.2±0.2(5)
26.0＊＊
10.6±0.1(5)
45.2＊＊ 152±3(4)
121±3(5)
20.4＊＊
103±2(5)
32.3＊＊
Xianyou ,1a 0.58±0.04(4) 1.34±0.06(5)
131.0＊＊
1.22±0.21(5)
110.3＊＊ 8.0±0.3(4)
9.4±0.1(5)
17.5＊＊
10.9±0.2(5)
36.3＊＊ 164±2(4)
135±2(5)
17.7＊＊
112±2(5)
31.7＊＊
Xianyou ,2a 0.66±0.05(4) 1.63±0.11(5)
147.0＊＊
1.33±0.03(5)
101.5＊＊ 3.8±0.1(4)
5.8±0.1(5)
52.6＊＊
8.6±0.2(5)
126.3＊＊ 198±2(4)
166±2(5)
16.2＊＊
140±2(5)
29.3＊＊
Note:The figure inside the parentheses denotes the number of example wi th the t reatmen t du ration of 24 hours;the data under
the line are increase ratio(%).t 0.01(5)=4.032, t 0.01(4)=4.604;t0.05(5)=2.571, t 0.05(4)=2.776;＊:P<0.05;＊＊:P<0.01.
2.2 Effects of the low-temperature treatment duration on contents of chlorophyll and proline
The results show the determination of contents of chlorophyll and proline in the different treatments
under low temperatures of 2 a seedling s of Fukiencypress in winter of 1999(Fig.1).
F ig.1 Varations in chlorophyll and proline in leaves of Fukiencypress under the low-temperature
It can be seen from Fig.1 that , af ter being t reated under -8 ℃ and -15 ℃fo r 24 hours , contents of
chlorophyll in seedlings of Fukiencypress increase remarkably , begin to low er after being t reated fo r 48
hours and approach the level at the natural low temperature(-4 ℃)when being treated for 72 hours w ith
an abrupt drop at -15 ℃.
Contents of proline increase w ith the increase of treatment duration and become the highest at durat ion
of 72 hours.It can be seen that , af ter the t reatment at -8 ℃ for 72 hours , the increase of proline is espe-
cially marked.
2.3 Effects of the low-temperature treatment duration on contents of soluble sugar and MDA
Fig 2 show s the results of determinat ion of contents of soluble sugar and MDA in the dif ferent t reat-
ments under low temperatures of 2 a seedlings of Fukiencypress in winter of 1999.
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南 京 林 业 大 学 学 报(自 然 科 学 版) 第 26 卷 第 5 期
Fig.2 Varations in contents of soluble sugar and MDA under the low-temperature treatment
◆——— -8 ℃;□——— -15 ℃
2.4 Relationship between the variations of physiological indices resulting from the low temperature
stressed treatment and the freeze injury
The synchronous observations made in the f ield show that the young seedlings of Fukiencypress have a
certain deg ree of f reeze resistance.The 1a seedlings suffered the slightly freeze injury under the natural low
temperature at -4 ℃fo r 24 hours w ith the f rozen ratio of 65.2%.Those w hich suffered the slight ly f reez-
ing were all phyllades of the primary and secondary branches on the side top , and wi thering due to f reezing
on the top w as not observed.The 2 a seedlings only encountered the slightly freezing of the phyllades on
some side branch t ips under the low temperatures of -4 ℃ for 48 hours in the field[ 8] .In w inter of 1999
when the natural low temperatures of -8 ℃lasted for 48 hours , the 2 a seedling s of Fukiencypress suffered
freezing , and the f rozen ratio w as 70%w ith tips of major seedlings no t being damaged due to f reezing , while
the 2 a seedlings all wi thered to death af ter undergoing the low-temperature treatment of -15 ℃fo r 48 and
72 hours;and 80% of the seedling s f roze to death af ter suf fering the low-temperature treatment at -8 ℃
for 72 hours.Though the above-mentioned result has no direct linear relationship w ith the detemined physi-
ological indices , it is possible to show the marked regularity .That is to say , under the low-temperature
stressed t reatment , contents of chlorophy ll , proline , soluble sugar and MDA in seedling s of Fukiencypress
w ould increase in different degrees w ith low ering of temperature w ithin a certain range.Besides , the roo t vi-
tality w ould also drop in varying deg rees;the low er the temperature , the quicker the drop.The low tempera-
ture of -8 ℃ and -15 ℃ for 24 hours didn t cause serious f reeze injuries on the seedling s.However , the
sustained treatment for 48 and 72 hours led to the marked drop in chlorophyll and increase in proline , solu-
ble sugar and MDA;moreover , the seedlings already suf fered serious freeze injuries.
3 Discussion
Chlorophy ll is an important substance fo r pho tosynthesis of the plants.In case of the low-temperature
stress ,most plants w ill display the marked drop in the ratio of photosynthesis;but there are also some excep-
tions.For instance , the ratio of photosynthesis of the betula on high mountains is greater than that of the
same provenance g row ing in the low-lying land[ 7] .The increase in chlo rophy ll w hich happened in this exper-
iment in Fukiencypress under the st ress of freeze also demonst rates that Fukiencypress still has the capabili-
ty of pho tosynthesis in a certain range under the low temperatures , thereby exhibi ting a kind of resistance
against the freeze injuries.
Infilt ration regulation is a kind of regulating mode of plants occurring under the stress of adverse cir-
cumstances.The regulating process is completed through cell biological synthesis and absorptive accumula-
tion of certain substances.Under adverse circumstances , plants mainly accumulate neutral amino acid , be-
taine , soluble sugar and some other organic substances.Among the neutral amino acids , the inf iltrat ion regu-
lation of proline is the most ef fective , and it will be accumulated in plants under w hatever adverse circum-
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2002年 总第 101期 HE Kai-yue et al:A study on variations of physiological indices of cold resistance in Fukiencypress
stancesare
[ 8] .Soluble sugar may both decrease icing point through infilt ration regulation and avoid cytoplasm
over dehydration so as to prevent cy toplasm colloid from clot ting w hen the plant encounters cold.So , the in-
crease of contents of soluble sugar may become a index of plant cold resistance.The present experiment
showed that the proline and soluble sugar in Fukiencypress increased in vary ing degrees under the low-tem-
perature t reatment ,which showed that this mode of regulation w as functioning .
The method adopted in the present experiment for determination of root vitali ty is theα-naphthylamine
method.Two Japanese named Aima and Shoka hold the opinion that the essence of oxidation ofα-naphthy l-
amine is cataly sis of pero xidase;the stronger the vitality of this enzyme is , the stronger its capability of o xi-
dation to α-naphthy lamineis.The peroxidase belongs to a kind of protective enzymes within plants , and i ts
action is mainly to get rid of the pero xide in bodies.It plays a very important role in resistance of plants.Af-
ter the low-temperature t reatment of Fukiencypress , the root vitality was found to have dropped to a certain
ex tent , which demonst rated that the activity of the peroxidase in bodies low ered.
MDA is a kind of poisonous substance in cell.In a normal temperature there is a lit tle MDA inside the
plant body.But w hen plant encounters the adverse environment , the content of MDA will increase.It can
dest roy the system of cell membrane.So , it is a symptom when plant cell is injured.The experimental result
showed that in a period of low-temperature the contents of MDA in Fukiencypress all increased.That is to
say , the cell membrane of plant w as perhaps dest royed.Moreover , the plant could not resto re to a normal
state.
The experiments found that , for the provenance of Xianyou , t reated at -8 ℃ and -15 ℃ for
24 hours , the increase in chlorophyll and proline w as relatively high;and the drop in roo t vi tality w as rela-
tively less(particularly for the 2 a seedlings).The observations made in the field also showed that only
slightly freeze injuries occurred when the low temperature of -8 ℃ lasted for 48 hours ,which showed that
i ts f reeze resistance w as relatively high , thereby possessing po tentiality for selection , improvement and ex-
pansion cultivation in other places.
[ References]
[ 1] Gao Zhaow ei.Rare t ree provenance for the use of timber———Fukiencypress[ J] .Fujian Science and Technology of Forestry , 1994 , 21(2):62
-66.
[ 2] Yang Zongw u ,Zheng Renhua ,Hou Boxing , et al.Rare tree provenance Fukiencypress[ J] .News Report of Forestry S cience and Technolo-
gy , 1988 ,(7):21-22.
[ 3] Xiao Xiangxi , Yang Zongwu , Ye Zhonghua , et al.Comparative analyses of the textures of the t imbers of Fukiencypress , China f ir and the ar-
t ificial forest of Masson s pine[ J] .New s Report of Forestry S cience and Technology , 2000 ,(2):3-5.
[ 4] Li Xiaochu , Huang Libin , Yang Zongwu , et al.Preliminary report of the seedling studies for the migrated cultivation of Fukiencypress[ J] .
Jiangsu Forest ry Science and Technology , 1999 , 27(3):1-4.
[ 5] Zou Qi.Experimental guide for Plant physiology and biochemistry[ M] .Beijing:China Agriculture Press., 1995.
[ 6] Zhang Zhiliang.Experimental guide for plant physiology[ M 〗.Beijing:Higher Educat ion Press.1995.
[ 7] He Jie , Liou Hongxian ,Wang Yirou , et al.Low temperature and photosynthesis of plants[ J] .Plant Physiology Communications.1986 ,(2):
1-6.
[ 8] Wang Hongchun.St ructural function and penetration regulation of biological membrane[ M] .Shanghai:Shanghai S cience and Technology
Press , 1987.
(责任编辑 是栋荣)
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南 京 林 业 大 学 学 报(自 然 科 学 版) 第 26 卷 第 5 期