全 文 ：EfectofColdStressonSomePhysiological
IndexesofPlantsinMalvaceae
WANGYu-qin1 , LIJian1, 2 , TIANHao-ren3 , JIANGChang-hua1, 2 , HUYong-hong1, 2＊
1.ShanghaiBotanicalGarden, Shanghai200231;2.ShanghaiEngineeringResearchCenterofSustainablePlantInnovation, Shanghai
200231;3.DepartmentofPharmaceuticalScience, NanchangUniversity, Nanchang330006
Abstract　ThepaperwastostudythechangetrendofthephysiologicalindexesinplantsofMalvaceaeundernormalandlowtemperaturecondi-
tion.TheresultshowedthattheincreaseextentofelectricalconductivityofplantswithcoldtoleranceinMalvaceaewassignificantlylowerthan
theplantwithoutcoldtoleranceunderthecoldstresscondition, whiletheirprolinecontentsandsolubleproteincontentswerealhigherthanthe
plantswithoutcoldtolerance.TheabovephysiologicalindicatorsalcanbeusedforthescreeningofvarietieswithcoldtoleranceinMalvaceae,
whichwilalsoprovidetheoreticalguidanceforthecoldtolerancescreeningofothergardenplants.
Keywords　Malvaceae;Coldstress;Physiologicalindexes
Received:December8, 2010　　Accepted:January10, 2011
SupportedbyResearchProjectinShanghaiGreen(Forest)Authori-
ty(G060317).
＊Correspondingauthor.E-mail:051023046@fudan.edu.cn
　　Malvaceaeplantshavenumerousspeciesandwidedistri-
butioninnature, includingbothpersistentrootherbsandtal
shrubs, andtheynotonlyareornamental, butalsocanbe
usedasindustrialrawmaterials.Theplantscanimprovethe
urbanecologicalconditions, andsimultaneouslymeetthebio-
logicaldiversity, sotheyhavegreatvalue.However, manyof
theplantsinthefamilyareheat-lovingspecies, andtheyhave
beenseriouslyinjuredinwinterinShanghairegion.Thus, in
ordertoscreenthecold-resistantvarietiesinthefamily, itis
veryimportanttostudythecoldtoleranceindicatorsofthe
plants.Currently, therehavebeennoreportsaboutthecold
toleranceindicatorsoftheplantsinMalvaceae.Tothisend,
accordingtothephenologicalrecordsofcoldtolerancetraits
oftheplantsinthefamily, theauthorpreliminarilyscreened
sixvarietieswithobviousdiferenceincoldtolerancetocarry
outcoldstress, anddetectedtheirrelevantphysiologicalindi-
cators, soastoclearthecorrelationbetweentherelevant
physiologicalindicatorsandtheircoldtolerancetraits, andfur-
therprovidetheoreticalguidanceforthecoldtolerancescreen-
ingofplantsinthefamilyandothergardenplants.
MaterialsandMethods
Materials
SixrepresentativevarietiesinMalvaceaewereselected
forthetest, andthebrancheswiththesimilarmaturitydegree
werecutduringthevigorousgrowthperiodinspring(room
temperature), andusedasthematerialsforthetest.Thede-
tailedinformationofthematerialsforthetestwasshownin
Table1.
Methods
Theelectricalconductivity, prolinecontentandtotalsolu-
bleproteincontentoftheplantsweredetectedundernormal
temperatureand-2℃/3 hcoldstresscondition, respectively.
Detectionofelectricalconductivity　Thesampleswere
takenusingholepuncherwiththediameterof1 cm(didnot
takemiddlevein).After0.2 gleaveswereweighedand
placedintotesttubes, 20 mldeionizedwaterwasaddedinto
thetubes, andvacuummachinewasusedtopumpaltheair
outoftubesfor15min.Thenthetubeswerestilyplacedun-
derroomtemperaturefor1h(slightlyshakingonceevery15
min), finalyDDS-11Atypeelectricalconductivitymeterwas
usedtodetecttheconductancevalue.Thetubeswereplaced
inboilingwaterbathfor10min, thentheyweretakenoutand
cooledtoroomtemperature.Theirconductancevalueswere
measuredafterslightlyshaking, andtherelativeelectrical
conductivitywerecalculated.Therelativeelectricalconductivi-
tywasexpressedbytheconductancevaluepergram fresh
weight(%/gFW).
Table1　Testvarieties
No.of
varieties Name
Coldtolerance
infield
1 Hibiscussyriacus Strong
2 Hibiscushamabo Strong
3 Hibiscusmutabilis` Jinbiyu Relativelystrong
4 Hibiscusmutabilis` PinkPeony Relativelystrong
5 Hibiscusmutabilis` WhitePeony Weak
6 Hibiscusmutabilis` Jinrui Weak
Detectionofprolinecontent　Theprolinecontentwas
measuredaccordingtothemethodbyZHANGDian-zhonget
al.[1] (1990).0.5 gsampleswereweighedandfulymiled
with5ml3%(W/V)sulfosalicylicacidsolutionforextraction.
Aftertheextractsolutionsweretransferredintotubes, they
wereplacedinboilingwaterbathfor10 min, andcentrifuged
at3 000 r/minfor10 min.0.2 mlsupernatantwasabsorbed
andboiledwith2.5% acidNinhydrinfor60 min.Aftertheso-
lutionwascooled, toluenewasusedtoextracttheredsub-
stances.Theextractionwasdetectedatthewavelengthof
520 nm, andprolinecontentwascalculatedaccordingtothe
standardcurve.
Detectionoftotalsolubleproteincontent　0.3 gleaves
wereweighed, addedwithextractionbufer(50 mmol/LTris-
HCl, 5 mmol/LMgCl2 , 1 mmol/LDTT, 0.2 mmol/LEGTA
pH7.5)accordingtotheratioof1∶3, andfulymiledinice
bath, thencentrifugedat27 000 ×gunder4 ℃ for15 min
twice.Thesupernatantwasabsorbedforlateruse.Theprotein
contentwasmeasuredbyCoomassiebriliantbluestaining
method, andthesampleconcentrationwasconfirmedaccording
PlantPhysiologyandBiochemistryAgriculturalScience＆Technology, 2010, 11(11-12):68-69, 123Copyright 2010, InformationInstituteofHAAS.Alrightsreserved.
toproteinstandardcurvedevelopedbyBradfordmethod.
ResultsandAnalysis
Detectionofelectricalconductivity
Coldstressdamagedthemembrane, thusthepermeabil-
ityofplasmamembranealsoincreased, andtheintracelular
protoplasmseepedout, resultingintheincreasedelectrical
conductivity.Theelectricalconductivityhadnegativecorrela-
tionwiththecoldtoleranceofvarieties[ 2] .ThedatainTable2
showedthattheincreasevalueofelectricalconductivityofHi-
biscusmutabilis`WhitePeony andHibiscusmutabilis`Jin-
rui withweakcoldtoleranceaftercoldstresswassignificantly
higherthanothervarietieswithcoldtolerance.
Table2　Changesofrelativeelectricalconductivity %/g· FW
No.ofvarieties Roomtemperature∥℃ Coldstress Increaseextent
1 78.3±5.1 126.4±4.6 48.1
2 88.5±2.1 137.8±3.2 49.3
3 105.7±6.4 157.3±3.0 51.6
4 104.4±3.3 165.7±10.0 61.3
5 108.7±10.1 209.2±2.3 100.5
6 102.5±4.2 220.3±3.5 117.8
Table3　Changesofprolinecontent μg/g· FW
No.ofvarieties Roomtemperature∥℃ Coldstress Increaseextent
1 26.0±1.0 132.2±8.1 106.2
2 16.6±3.1 121.4±3.3 102.8
3 24.5±3.7 122.6±6.7 98.1
4 27.4±2.2 131.4±7.2 99.2
5 21.2±1.1 85.5±4.5 64.3
6 21.2±1.4 88.6±5.9 67.4
Prolinecontent
Freepralineisoneofthemainsubstancesforosmotic
adjustmentwithincels.Theprolinecontentofplantsinnormal
circumstancesisverylow, whichwilbeincreaseddramatical-
lyinadversity.Theprolinecontentwilbeincreasedrapidly
undercoldstress, sotheanti-dehydrationofcelsisalsoen-
hanced, andtheenhancementofprolineunderstressisaver-
ysensitiveindicator[2] .AsshowninTable4, theincreaseex-
tentofprolinecontentincoldtolerancevarietieswassignifi-
cantlyhigherthanthatinthevarietieswithoutcoldintolerance.
Therefore, theprolinecontentisoneoftheidealindicatorsfor
thecold-resistancedetectionofvarietiesinMalvaceae.
Table4　Changesoftotalsolubleproteincontent mg/g· FW
No.ofvarieties Roomtemperature∥℃ Coldstress Increaseextent
1 4.86±0.33 6.18±0.31 1.32
2 4.48±0.11 5.79±0.33 1.31
3 4.76±0.24 5.52±0.06 0.86
4 4.45±0.08 4.76±0.12 0.31
5 4.70±0.13 3.87±0.06 -0.83
6 4.76±4.2 4.24±0.11 -0.52
Totalsolubleproteincontent
Thesolubleproteincontentofplantsunderlowtempera-
turehascorrelationwiththeircoldtolerance[ 3] .Asshownin
Table4, thesolubleproteincontentincold-resistantvarieties
wasenhanced, whilethecontentinvarietieswithoutcoldtol-
erancewasdecreased, thisindicatedthatsolubleproteincon-
tentwasconsistentwithcold-resistantcapacityofvarieties.
Therefore, solubleproteincontentcanbeusedasoneofthe
indicatorsfortheidentificationofcold-resistantvarietiesin
Malvaceae.
Discussion
Freezinginjurymainlydestructsthecelmembraneoftis-
sue, thuscausingthedysfunctionintheaspectsofphysiolo-
gy, metabolismandbiochemistry.Whenplantsareunderlow
temperaturestress, theunsaturatedfatyacidofcelmem-
branewilhaveoxidativedegradation, thusthegapsbetween
membranesarelarger, resultingintheleakageoflargenum-
berofintracelularion, sotheelectricalconductivityoftissue
exudatesisalsoincreased.Therefore, throughthechanges
ofelectricalconductivityoftissueexudatestodeterminethe
injuryconditionofcelsisthemostcommonlyusedmethodfor
determinationofcoldtolerance[2] .
Prolinecontentisasensitivephysiologicalindicatorfor
characterizationofcoldtolerance.Thefreepralinehasclose
relationshipwithwaterpotentialofplantcels.Ifprolinecon-
tentishigher, indicatingthatwaterholdingcapacityofthe
celsisalsoenhanced.Studieshaveshownthatafterlow
temperaturestress, theinternalfreeprolinecontentsinplants
wilberapidlyincreased.Freepralinehasthecharacteristics
ofwatersoluble, highwaterpotentialandnon-toxicityafterin-
tracelularaccumulation, thusitcanbeusedasanti-dehydra-
tingagenttoprotectplantsduringtheprocessoflowtempera-
turestress, itcanalsoinducethecoldtoleranceofplants[ 2] .
Thevarietieswithoutcoldtolerancewilbegintoproduce
heatshockproteinsintheinitialphaseofcoldstress;while
thevarietieswithcoldtolerancehaverelativelyhinderre-
sponsetolowtemperaturestress.Whenthestresstimeis
prolonged, theproteincontentofthevarietieswithcoldtoler-
anceisincreased, whilethesynthesisofheatshockproteinin
thevarietieswithoutcoldtoleranceisdecreased, showingthe
decreaseoftotalsolubleproteincontent[ 3] .
Thechangetrendoftheabovephysiologicalandbio-
chemicalindicatorsisconsistentwiththecoldtoleranceof
plantsinMalvaceaeunderroomtemperatureandcoldstress
processes, sotheindicatorscanbeusedforthescreeningof
varietieswithcoldtoleranceinMalvaceae.Theresultcanalso
beusedasthescreeningindicatorofcoldtoleranceforother
gardenplants, whichwilprovidefoundationfortheestablish-
mentofphysiologicalindicatorsystem forthescreeningof
coldtoleranceofgardenplants.
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云南铁壳麦变种分类及基于农艺性状的遗传多样性分析
王志伟 ,杨金华 ,程加省 ,胡银星 ,程耿 ,于亚雄＊　(云南省农业科学院粮食作物研究所 ,云南昆明 650205)
[目的 ]为给云南省小麦新品种选育以及物种保护研究提供依据。
[方法 ]对 29份云南铁壳麦进行了变种分类和基于 14个农艺性状的遗传多样性分析。
[结果 ]云南铁壳麦 A14为一未定名的白粒变种 ,其余 28份材料分为 10个已定名变种类型;云南铁壳麦农艺性状变异较丰富 ,以不孕小穗数
变异系数最大(22.59%),抽穗期的变异最小;云南铁壳麦 7个农艺性状的多样性指数变化较大 ,为 1.55～ 2.04。 29份云南铁壳麦分为 3个
类群 ,但同一变种类型的云南铁壳麦并未整齐的聚为 1类;云南铁壳麦A13、A14和A21间的遗传关系相对较近 ,而与其他云南铁壳麦间的遗
传关系相对较远。
[结论 ]云南铁壳麦在农艺性状上存在较为广泛的遗传多样性。
关键词　云南铁壳麦;农艺性状;遗传多样性
基金项目　云南省重点基金(2008CD010);国家自然科学基金(30760119);云南省麦类科技攻关 UQEC资助项目(2010BB005)。
作者简介　王志伟(1985-),男 ,湖北松滋人 ,研究实习员 ,硕士 ,从事小麦遗传育种研究 E-mail:wzw0923@ 126.com。 ＊通讯作者 ,研究员 ,硕士
生导师 ,从事小麦遗传育种研究 , E-mail:yyx582@vip.sohu.com。
收稿日期　 2010-12-02　　修回日期　 2010-12-27
(上接第 69页)
冷胁迫对锦葵科植物部分生理指标的影响研究
王玉勤 1 , 李 健 1, 2 ,田浩人 3 ,蒋昌华 1, 2 ,胡永红1, 2＊　(1.上海植物园 ,上海 200231;2.上海城市植物资源开发应用工程技
术研究中心 ,上海 200231;3.南昌大学药学系 ,江西南昌 330006)
摘要　锦葵科植物研究正常温度和低温条件下锦葵科植物品种部分耐冷生理生化指标的变化趋势。结果表明锦葵科植物耐冷品种在冷胁
迫条件下电导率上升明显小于不耐冷品种 ,而脯氨酸含量、可溶性蛋白含量均比不耐热品种高。上述生理指标均可作为锦葵科植物品种耐
冷性筛选指标 ,也为其他园林植物的耐冷筛选提供理论指导。
关键词　锦葵科;冷胁迫;生理指标
基金项目　上海市绿化(林业)管理局科研项目(G060317)。
作者简介　王玉勤(1963-),男 ,上海人,高级工程师 ,从事引种驯化研究。 ＊通讯作者, E-mail:051023046@fudan.edu.cn
收稿日期　 2010-12-08　　修回日期　 2011-01-10
123
WANGZhi-weietal.MutationClassificationandGeneticDiversityAnalysisofTriticumaestivumssp.yunnanenseKingBasedon
AgronomicTraits