全 文 ：武汉植物学研究 2001, 19( 5) : 421～427
Journal of Wuhan Botanical Research
Cissus quadrangularis L.及其它
小型肉质植物标本制作中的压制前处理

GITURU Wahiti Robert　王青锋** 　王　勇　郭友好
(武汉大学生命科学学院,武汉　430072)
摘　要: 以Cissus quadrangular is L . 为材料用不同的技术处理并比较其收缩程度、色彩保真
程度以及茎附属物的保留情况。试验了 3 种常规预处理方法及 2 种针对肉质组织的处理方
法, 并进行 ANOVA( Ana ly sis of va riance)分析,结果显示 3种常规植物标本预处理方法明显
不同(P < 0. 005) , 用 LSD( least significant difference)分析方法分析得出深冷冻法对植物标
本造成的损害比汽油法( LSD= 1. 708)和福尔马林法( LSD= 2. 065) 要小;汽油法和福尔马林
法处理后得到的标本质量无明显差别。用 ANOVA 法分析醋酸法、纵切法得到的标本及未经
任何处理的标本, 它们的效果也有明显不同( P< 0. 001) ,醋酸法得到的标本比纵切法 ( LSD
= 2. 371) 和未经任何处理得到的标本 ( LSD= 2. 138) 效果更好 ,而纵切法和未经任何处理
得到的标本并无明显不同( LSD= 0. 233NS)。醋酸处理法制作小型肉质植物标本其效果得到
了植物标本馆工作者的肯定。
关键词: Cissus quadrangular is; 标本馆技术; 肉质植物
　　中图分类号: Q94-34+ 2　文献标识码: A　文章编号: 1000-470X ( 2001) 05-0421-07
Pre-Pressing Treatment for Cissus quadrangularis L. and
Other Small Succulents
GIT URU Wahit i Robert , WANG Qing-Feng

, WANG Yong , GUO You-Hao
( Col leg e of L if e S ci ences, W uhan Unive rsity, Wuhan　430072, Ch ina)
Abstract: Herbarium specimens o f Cissus quadrangularis L. w ere prepared by
dif ferent techniques and compared for shr inkage, retent ion of color, and persi-
stence of stem appendages. The ef ficacy of thr ee pret reatment techniques and
tw o methods o f handling succulent t issue were invest igated. Using analysis o f
variance ( ANOVA) there was show n to be a signif icant dif ference ( P< 0. 005)
betw een the three pret reatment techniques. U sing the least significant dif fe-
rence( LSD) test, the deep- freezing technique w as shown to produce less dis-
to rt ion in the r esultant specimens than the gaso line technique ( LSD = 1. 708)




作者简介: GITU RU Wah iti Rob ert ( 1967- ) ,男,肯尼亚人,博士,从事系统与进化植物学研究。通讯作者。
收稿日期: 2000-12-04,修回日期: 2001-04-19。
基金项目:国家自然科学基金资助项目(项目号: 30070055, 39770055)。
and the formalin technique ( LSD= 2. 065) . No signif icant difference w as ob-
served in the quality o f specimens pr oduced by the formalin and the gasoline
techniques ( LSD= 0. 357N S) . A NOVA test indicated a signif icant difference
in eff icacy betw een the acet ic acid t reatment , the longitudinal sect ioning t reat-
ment and untreated specimens ( P< 0. 001) . T he acet ic acid pretr eatment w as
show n to be superior to the longitudinal sect ioning pret reatment ( LSD =
2. 371) and untreated specimens ( LSD= 2. 138) . No signif icant dif ference w as
found betw een the untreated specimens and those subjected to long itudinal sec-
tioning ( LSD = 0. 233NS ) . The acet ic acid pret reatment technique is r ecom-
mended for t reat ing specimens of small succulents by herbarium wor kers.
Key words : Cissus quadrangularis; Her barium technique; Succulent plants
Good her barium specimens of succulent plants cannot be prepared by apply ing ordi-
nary drying methods w ithout some special t reatment [ 1] . Special techniques are r equired
in o rder to preserve the shapes of specimens as far as possible and to make them dry rea-
sonably rapidly. T he autho rs have exper ienced considerable diff icult ies in the prepara-
tion o f herbarium specimens of succulent plants. T hey r ecognized a need for bet ter
methods bo th for the speedy killing o f the plant material to stop gr ow th while in the
pr ess and for pr e-pressing techniques that deal w ith the excessive succulent t issue. If
these pro cedures are not followed in drying succulents, the stems stay aliv e in the pr ess
fo r many months, and in the process, abscission layers fo rm and cause the leaves, f low-
er s and fruits as w ell as o ther stem appendages to fall of f . Eventually fung i at tack the
specimens and they blacken w ith mo ld and f inally rot .
Improved methods designed to produce bet ter specimens have been suggested
[ 2 5] .
Dif ferent methods have been used for killing specimens of succulent plants pr io r to
pr essing w ith different degrees of success. Invest ig at ions are requir ed to identify the
most appropriate method of kill ing succulent plant materials. The desirable method
should result in herbarium specimens w ith lit t le disto rt ion and adequate retent ion of the
natural colo rs of the plant parts. Excessive detachment o f stem appendages such as ten-
drils and flowers must also be avoided.
Logan
[ 6]
developed a pre-pressing t reatment technique that w as found to be suitable
in hastening dr ying of specimens of Begonia species and some succulents. T he technique
involves so aking the specimens in a dilute solut ion of acet ic acid laced w ith a few drops
of deter gent . This technique has not yet been tested on C. quadr angularis. It is also
necessary to compare the herbarium specimens obtained by this technique w ith those ob-
tained from material prepared by the convent ional methods used for specimens of C.
quadrangularis.
1　Materials and methods
1. 1　Fieldwork
Localit ies in Kenya for specimens o f C. quadrangularis reco rded in the East African
422 武汉 植 物学 研究　　　　　　　　　　　　　　　　第 19卷　　
herbarium, Nair obi ( EA) w ere visited. The populations of the species w ere observed in
the wild, and living material was collected. Relevant information that g oes with a
herbarium specimen including the alt itude, grow th habit, co lor and scent of flow ers,
co lor of leaves and stems, and the color of y oung and ripe f ruit s w as recor ded.
1. 2　Voucher specimens
Materials for voucher specimens w as preser ved in tw o w ays, namely as spirit col-
lect ions and as pressed specimens. Spirit collect ions w ere prepared by storing specimens
in a m ix tur e of formaldehyde-alcoho l-acet ic acid ( FAA) in the rat io of 3
6
3. Pressed
specimens w ere prepared follow ing the methods of Forman and Br idson
[ 7] . Voucher
specimens in this study w ere deposited in the Kenyat ta U niversity herbarium and the
Wuhan U niversity herbarium ( WH) .
1. 3　Specimen pretreatment techniques
1. 3. 1　Killing plant specimens
To compare the differ ent pr et reatment methods used for killing plant specimens be-
fo re pr essing, the follow ing pr ocedure w as adopted.
Dimensions of v arious plant o rgans w er e measured and recorded fr om thirty speci-
mens of C. quad rangularis. M easurements w er e made with a sensitive vernier calliper
w ith a sensitivity of 0. 002 mm . T his w as done immediately af ter gathering to ensure
that the material was st ill f resh and minimize er rors arising f rom shrinkage. T he dimen-
sions measur ed included: stem ( w idth of one face) , node ( w idth o f one face) , internode
leng th, tendr il diameter, length o f leaves and width of leaves.
( a ) T en specimens were placed in the fr eezing compartment o f a ref rigerator for 2
hour s and then removed and allow ed to thaw .
( b) T en specimens w ere dipped in pet rol for about 12 hours and then removed and
the pet rol was allowed to evaporate.
( c) Ten specimens w ere immersed in 40% formalin for about 2 hours and then r e-
moved and the formalin w as allow ed to evaporate.
( d) The specimens w ere placed individually in plant presses. The blo tters and ab-
sor bent paper s w ere changed every day until no more damp spots w ere observ ed on the
blot ters. For the purposes of the experiment , the specimens were kept in the pr esses for
one month. T he specimens w er e observ ed and re-measured af ter the one-month per iod
to determine dimensional changes. They w ere also obser ved for retent ion of co lor and
pr esence o f detached pieces e. g. leaves and tendrils. The exper iment involved six t reat-
ments w ith each treatment having 10 replicates.
1. 3. 2　Dealing with succulent tissue
Measurements of org an dimensions w ere obtained from 30 specimens as in step 1. 3.
1 above. T he specimens w er e placed in the f reezing compartment of a refr ig er ato r for 2
hour s and then removed and allow ed to thaw .
423　第 5期　GIT URU W R 等: Cissus quadr angularis L.及其它小型肉质植物标本制作中的压制前处理(英)
( a) Ten of the specimens w ere soaked for 48 hour s in 2 liter s of 5% acet ic acid into
w hich four drops o f deterg ent had been added.
( b) Ten of the specimens had their stems cut long itudinally to cut through the wa-
terproo f cuticle.
( c) Al l the thirty specimens w ere placed indiv idually in plant presses. T he blot ters
and absorbent papers were changed every day and, for each specimen, the number o f
day s taken before damp spots disappeared from the blot ters w as reco rded. For the pur-
poses o f the exper iment , the specimens w ere kept in the pr esses for one month. T he
specimens w ere observ ed and re-measured af ter the one -month per iod to determine the
dimensional changes. They w ere also observed for retent ion of color and presence of de-
tached pieces e. g . leaves and tendrils . The experiment involved six t reatments w ith
each treatment having 10 r eplicates.
2　Results
2. 1　Pretreatment techniques
The resul ts o f dimensional changes occur ring in the herbarium specimens prepared
fr om specimens of C. quadrangulari s killed by three dif ferent methods are summarized
in tables 1 and 2.
表 1　3种不同的固定方法下 10份 C. quadrangu-
laris 标本的器官尺寸平均变化百分数和干燥所需
的平均天数(这些标本在处理之后压制了 1 个月)
T able 1　Mean percent age or gan dimension
changes and mean time required fo r dry ing ( days )
of 10 specimens o f C . quadrang ularis
Organ dimens ion
Pr et reatment
Deep freez e Gasoline Formalin
S tem diameter 2. 89 20. 68 17. 31
Node diameter 3. 56 11. 53 13. 63
Internode length 0. 60 3. 05 3. 40
T endril diameter 3. 08 31. 82 32. 29
L eaf length 21. 65 29. 81 21. 61
L eaf w idth 19. 36 23. 53 37. 94
Drying t ime( d) 9. 8 10. 1 10. 6
表 2　3 种不同预处理方法下的 ANOVA 表
T able 2　Analysis o f var iance( ANOVA) t able fo r
compar ison betw een thr ee pr etreat ment techniques
Sour ces of
variat ion
Sums of
squares
Degrees of
fr eed om
Mean sum s
of squares
F
T reatment 216. 435 2 108. 218 83. 502
Organs 14. 623 5 2. 925 2. 257
Error 14. 257 11 1. 296
T otal 18
T reatments: T abular value at degrees of fr eedom 　
11, 2= 13. 81; P< 0. 001* ** .
Organs: T abu lar value at degrees of f reedom 　
11, 5= 3. 20; P > 0. 05.
LSD: A. Freezing meth od= 2. 142, A- B= 1. 708** ;
B. Gasol ine meth od= 3. 85, B- C= 0. 357 NS;
C. Formalin meth od= 4. 207, A- C= 2. 065** .
2. 2　Specimen shrinkage and appendage retention
The results of dimensional changes occurring in herbarium specimens prepared fr om
specimens of C. quadrangularis subjected to tw o differ ent specimen pr et reatment tech-
niques as w ell as f rom untreated specimens are summarized in tables 3 and 4. Figure 1
( A ) show s a r epresentat ive specimen treated by deep freezing at - 4℃ for 2 hours be-
fo re pressing . F igure 1( B) show s a representat ive specimen subjected to the same pr e-
pr essing tr eatment as specimen ( Fig . 1: A ) follow ed by soaking in 5% acet ic acid for 48
hour s before pressing. The fo rmer specimen is char acterized by extensive detachment o f
424 武汉 植 物学 研究　　　　　　　　　　　　　　　　第 19卷　　
appendages and discolor at ion of the inf lorescences. T he lat ter specimen display s marked-
ly superior retent ion of appendages.
图 1　A.用- 4℃低温处理 2 h 之后压制得到的一个具代表性的标本( Gituru 22) (×0. 4) (请注意
叶片及茎卷须的脱落和花序的褪色情况) ; B.用- 4℃低温处理 2 h之后再用 5%醋酸处理 48 h得
到的结果( Gituru 24) (×0. 6) (请注意叶片及茎卷须并未脱落)
Fig. 1　A. A representat ive sp ecim en of C . quad rangular is( Gitur u 22) t reated by deep f reezing at
- 4℃ for 2 hours before pres sing (× 0. 4) ( N B th e detachmen t of leaves and tendril s and dis co-
lor at ion of the inf lorescences ) ; B. Represen tat ive specimen of C . quad rangular is( Gituru 24) t reat-
ed by deep f reezing at - 4℃ for 2 hours follow ed by soak ing in 5% acet ic acid for 48 hours before
press ing(×0. 6) ( NB the pres ence of leaves an d tendrils w hich have not become detached )
　表 3　3种不同的预处理方法下 10 份 C. quadran-
gularis 的器官尺寸平均变化百分数和干燥所需的
平均天数(这些标本在处理之后压制了 1 个月)
T able 3　Mean percent age or gan dimension
changes and mean time required fo r dry ing ( days )
of 10 specimens o f C . quadrang ularis
Organ dimens ion
T echnique
Longitudinal
sect ionin g
Acet ic acid
t reatment
U nt reated
specimens
S tem diameter 32. 04 11. 28 16. 04
Node diameter 13. 53 7. 94 9. 07
Internode length 2. 97 0. 03 4. 05
T endril diam eter 28. 33 26. 59 33. 56
Leaf length 8. 37 1. 39 8. 82
Leaf w idth 21. 54 12. 54 22. 91
Drying t ime 7. 5 6. 8 9. 8
表 4　3 种用来处理肉质植物组织的方法比较后得
出的 ANOVA表
T able 4　ANOVA table fo r compar ison of 3 dif-
fer ent methods of dealing w ith succulent t issues
Sour ces of
variat ion
Sums of
squares
Degrees of
fr eed om
Mean sum s
of squares
F
T reatment 20. 486 2 10. 243 7. 786
Organs 117. 237 5 23. 447 17. 824
Error 13. 155 10 1. 315 5
T otal 17
T reatments: Tabular value at degrees of f reedom 　
10, 2 = 7. 786; P < 0. 05* .
Organs: T abu lar value at degrees of f reedom 　
10, 5 = 17. 824; P < 0. 001** * .
LSD: A. Longitudin al sect ioning A- B= 2. 371** ; B.
Acet ic acid pre-press ing t reatment A- C= 0. 233 NS ;
C. Unt reated specimens B- C= 2. 138** .
425　第 5期　GIT URU W R 等: Cissus quadr angularis L.及其它小型肉质植物标本制作中的压制前处理(英)
3　Discussion
In preparat ion o f her barium specimens of C. quadr angular is the method of killing
plant material by freezing has been found to pr oduce her barium specimens that are supe-
rior to those obtained from material killed by immersing in formalin or gasoline. Free-
zing resul ted in herbarium specimens w ith less distort ion. In addit ion, the specimens
obtained show ed greater r etent ion of natural co lors and less tendency o f detachment o f
pieces such as tendrils , leaves and f low ers.
It is common pract ice for some botanists to use formalin for killing specimens o f
succulent mater ial in the f ield before placing them in plant pr esses. This study has
show n that the use of gaso line eventually produces specimens that have less distor tion
than those obtained from specimens tr eated w ith formalin. T his f inding is especial ly im-
por tant since it could have ser ious implicat ions fo r field botanists w ho are f requently en-
cumber ed by having to car ry containers of FAA to the field. Gasoline would be more
readily available under these condit ions since it is carried as part o f the emergency sup-
plies of motor vehicles. In addit ion, g asoline is mo re commonly available than “odorless
carr ier”and unlike o/ c g asoline evapo rates r apidly f rom the plant mater ial . T his elimi-
nates the need to hang up material befo re pr essing which is necessary for material tr ea-
ted with o/ c. The vo latile highly inf lammable nature o f gasoline makes it necessary to
avoid w orking in closed places and near any sources of f ire. These condit ions, w hile not
being part icularly diff icult to achiev e, especially under f ield condit ions, present serious
limitations to the w idespread use of this technique. In spite of producing good results,
the use of microw aves for killing and dry ing specimens o f succulent plants is unlikely to
become w idespread in developing countries. T his is larg ely due to the high cost of the
equipment required for this technique.
The pre-pressing t reatment technique developed for B egonia species by Logan
[ 6]
has
been found to be ef fective for preparing specimens of C. quadrangularis and o ther small
succulents. A soaking period of 48 hours in tw o liters of 5% acet ic acid laced w ith 2
dr ops of detergent w as found to be satisfacto ry. Compar ed to untreated specimens as
w el l as those t reated by longitudinal sect ioning befo re pressing , the specimens subjected
to the acet ic acid t reatment dried faster , show ed less disto rtion and eventually retained
mor e natural co lor after pressing . The use of the acetic acid technique, in combination
w ith the technique o f killing specimens by f reezing , r esul ts in specimens of superior
quality and could reduce considerably the cost of preserving the specimens in her baria.
This is part icularly impo rtant in many developing countr ies w here herbaria have been
facing considerable budgetary const raints.
Acknowledgments: The author s appreciat ion goes to P ro fesso r L . E. New ton of the Depar tment of
Bo tany , Kenyatta U niver sity fo r his invaluable contr ibution to the study .
426 武汉 植 物学 研究　　　　　　　　　　　　　　　　第 19卷　　
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[ 2 ]　De Wolf J r G P. Notes on making a h erbarium . A rnold ia, 1968, 28: 69 111.
[ 3 ]　Smith C, Earle E. Prepar ing herbarium specimens of vas cular plants . Ag r Inf B ull U S Dept A gr i, 1971,
348: 710 716.
[ 4 ]　Jain S K, Rao R R. A Handbook of Field and Herbarium Methods . New Delhi: Today and T omorrow Printers
and Publ ishers, 1977.
[ 5 ]　Sanchez M erjorada H . S ucculentas . In : L ot , Ch iang eds. Manual Del H erbario. Kew : Royal Botanical Gar-
den s, 1986: 103 111.
[ 6 ]　Logan J . A pre-pr ess ing t reatment for B egonia species and su ccu len ts . T ax on, 1986, 35: 671.
[ 7 ]　For man L, Bridson D. T he Herbarium Handbook . Kew : Royal Botanical Garden s, 1989.
427　第 5期　GIT URU W R 等: Cissus quadr angularis L.及其它小型肉质植物标本制作中的压制前处理(英)