全 文 ：ClassificationStatusofXinjiangPearIdentifiedby
AFLP
LUFeng-juan＊
EnvironmentalManagementColegeofChina, Qinhuangdao066004
Abstract　[ Objective] TheaimwastoprovideDNAlevelbasisforXinjiangPearclassificationposition.[Method] Throughclusteranalysisand
geneticsimilaritycoeficientanalysis, theclassificationstudyonXinjiangPearwascariedoutbyusingAFLPmolecularmarkertechnique.[ Re-sult] Whenthethresholdvalueis15, XinjiangPearcultivarLanzhouchangbaholdtogetherwithHuachangbafirst, thenwithPyruscommunisL.
cultivarsBali, Hongbali, Hongqie, Qieli, Baoliasika, ZhulibienandXinjiangPearcultivarQiliamuti.Among7XijiangPearcultivars, theeuclide-
andistanceamongspecieswithingroupsrangedfrom2.646to10.050.AndthesmalesteuclideandistancebetweenXinjiangPearandP.pyrifo-
liaNakai, P.communisL, P.BretschneideriRehdaswelasP.ussuriensisMaximwere7.746, 7.746, 7.810and8.165, respectively.[Conclu-
sion] XinjiangPearhastheclosestrelationshipwithP.communisL.
Keywords　Molecularmarker;XinjiangPear;Classificationstatus;AFLP
Received:December4, 2009　　Accepted:January12, 2010
SupportedbyNaturalScience Foundation ofHebeiProvince
(302240).
＊Corespondingauthor.E-mail:lujuan0335@163.com
　　XinjiangPearareproducedinXinjiangProvince, Qinghai
Province, GansuProvince, etc..Theirfruitshapesaresimi-
lartothatofP.communisLexceptforlongfruitstalk, and
theirleavesarezigzagandhaveverylargemorphologicalvar-
iation.Therehavebeenmanyresearchesonthemorphology,
enzymologyandpalynologyofXinjiangPear, buttherehave
beendisputesontheclassificationstatusofXinjiangPear.In
ordertoprovideabasisfortheclassificationstatusofXinjiang
PearatDNAlevel, theauthormadeananalysison44 Pyrus
materialsbyAFLPmolecularmarkertechniquesoastoex-
ploretheclassificationstatusofXinjiangPearatDNAlevel
emphaticaly.
MaterialsandMethods
Testedmaterialsandreagents
Testedmaterials:44testedcultivars, amongwhichthere
were7XinjiangPearcultivars, wereprovidedbynationalpear
germplasmnursery(Xingcheng, Liaoning)ofResearchInsti-
tuteofPomology, theChineseAcademyofAgriculturalSci-
encesandthespecimengardenofHebeiAgriculturalUniversi-
ty(Table1).Reagents:MseIwasboughtfromNewEngland
BiolabsCompany;T4DNAligasewasboughtfromPromegaCompany;EcoRI, TaqDNApolymeraseanddNTPwere
boughtfromTaKaRaBiotechnology(Dalian)Co., Ltd.;Link-
ersandprimerswerealsynthesizedbyShanghaiSangonBi-
ologicalEngineeringTechnology＆ ServicesCo., Ltd.;And
theotherreagentswerealboughtfromBaodingChemicalRe-
agentsMarketingDepartment.
Methods
DNAextraction　DNAwasextractedfrommatureleavesof
newshootsofhealthyplantsbyCTABmethod[ 1-2].
AFLPanalysis　AccordingtothemethodofLuFengjuanet
al[ 3] , amplificationwascarriedoutbyusing10 screening
primercombinations(Table2).
Table1　44MaterialsofPyrusappliedinthisstudybyAFLP
No. Cultivars System No. Cultivars System
1 Wangziershishiji P.pyrifoliaNakai 23 Taihuangli P.BretschneideriRehd
2 Aiganshui P.pyrifoliaNakai 24 Jinhuali P.BretschneideriRehd
3 Xinxing P.pyrifoliaNakai 25 JinhuaNo.4 P.BretschneideriRehd
4 Jinxing P.pyrifoliaNakai 26 Pinguoli Unknown
5 Jincunqiu P.pyrifoliaNakai 27 Pingxiang Apple-pear×Yanbianxiehuatian
6 Bali P.communisL 28 Jinxiang Nanguoli×Bali
7 Hongbali P.communisL 29 ZhongliNo.1 Xinshiji×Zaosu
8 Hongqie P.communisL 30 Guifeili P.communisL
9 Baoliasika P.communisL 31 Tianshengfu P.BretschneideriRehd
10 Zhulibien P.communisL 32 Zaoshujuju XinjiangPear
11 Qiliamuti XinjiangPear 33 Aidang P.pyrifoliaNakai
12 Wuweibingzhuli XinjiangPear 34 Xuehua P.BretschneideriRehd
13 Huachangba XinjiangPear 35 Jinsu P.BretschneideriRehd
14 Lanzhouchangba XinjiangPear 36 Qinfeng P.BretschneideriRehd
15 Kuerlexiangli XinjiangPear 37 Huangxianchangba P.BretschneideriRehd
16 Nanguo P.ussuriensisMaxim 38 Ningmenghuang Jinbaili×P.communisL
17 Nanguoli P.ussuriensisMaxim 39 Qixiadaxiangshui P.BretschneideriRehd
18 Xiaoxiangshui P.ussuriensisMaxim 40 Huangsuanli XinjiangPear
19 Yali P.BretschneideriRehd 41 Ergongbai Yali×Zhenyu
20 Jinxiandayali P.BretschneideriRehd 42 Tietou Unknown
21 Zhaoxiandayali P.BretschneideriRehd 43 Jingbai P.ussuriensisMaxim
22 Chuizhiyali P.BretschneideriRehd 44 Qieli P.communisL
Resultrecording　Thestableandclearbondwasselected
fromamplifiedproductsofeachprimertobeasstatisticalda-
ta.Bondwasdenotedas1, andnobondwasdeno-
tedas0.Finaly, thepolymorphicstatisticaldatawasinput
AgriculturalScience＆Technology, 2010, 11(1):3-4, 30
Copyright 2010, InformationInstituteofHAAS.Alrightsreserved. AgriculturalBiotechnology
intocomputer.
Clusteranalysis　Euclideandistancebetweentestedmateri-
alswascalculatedbySPSSforWindows.Clusteranalysis
wascarriedoutbysquaresumofdeviationssoastooutput
euclideandistancelistandgeneratedendrogram.
Table2　Thescreeningprimersandtheirselectivebases
Primer
combination
Selective
bases
Primer
combination
Selective
bases
M2 /E5 M-CAG/E-ACG M5 /E6 M-CTC/E-AGCM2 /E6 M-CAG/E-AGC M6 /E5 M-CTG/E-ACGM2 /E7 M-CAG/E-AGG M6 /E7 M-CTG/E-AGGM3 /E8 M-CAT/E-ACT M7 /E5 M-CTT/E-ACGM5 /E4 M-CTC/E-ACC M7 /E6 M-CTT/E-AGC
ResultsandAnalysis
Clusterresults
Itwasfoundfromtheconstructedclustergraph(Fig.1)
of44 pearcultivarsthatwhenthresholdvaluewas15, Xin-
jiangPearcultivarsLanzhouchangbaandHuachangbaclus-
teredtogetherfirstly, andthenclusteredtogetherwithP.com-
munisLcultivarsBali, Hongbali, Hongqie, Qieli, Baoliasika
andZhulibienaswelasXinjiangPearcultivarQiliamuti.
Fig.1　DendrogramofsometestedPyrusmaterialsbasedon
AFLP
Euclideandistance
AccordingtotheresultsofAFLPamplification, itcouldbe
calculatedthatamongthe7 XinjiangPearcultivars, theeu-
clideandistancebetweencultivarswithinpopulationsliedbe-
tween2.646and10.050;andthesmalesteuclideandistance
betweenXinjiangPearandP.pyrifoliaNakai, P.communisL,
P.BretschneideriRehdaswelasP.ussuriensisMaximwere
7.746, 7.746, 7.810 and8.165 respectively, whichindicates
thatXinjiangPearhaveaclosergeneticrelationshipwith
P.pyrifoliaNakaiandP.communisL.
ConclusionsandDiscussion
Inthemiddlestageof1950s, professorYuDejunisolated
XinjiangPearfromotherpearcultivars, establishedtheinde-
pendentpositionofXinjiangPearinChinesePomological
Taxonomy, andheldthatXinjiangPearwasthehybridculti-
varofP.communisLandP.BretschneideriRehd[ 4] .In2001,
TengYuanetal[5] analyzed13 Pyruscultivarswhichwerepro-
ducedinChinaoriginaly, andexploredtheclassificationstatus
ofXinjiangPear.LiXiugenetal[ 6] madeafurtheranalysison
theorigin, evolutionandclassificationofPyrusinChinaby
polenmorphologyquantificationin2002, andthenfoundthat
therewasaclosergeneticrelationshipbetweenP.ussuriensis
MaximandXinjiangPear, whichindicatesthatthe3 systems
havethesameorsimilarparentingenetics.Kimuraetal[ 7]
madeaSSRanalysisonAsianpear(28 Japanesepearculti-
varsand19 Chinesepearcultivars)andCommonPearby
SSRlabelingtechnique, andthenfoundthattherewasnoob-
viousseparationinChinesepearcultivarsP.Bretschneideri
Rehd, P.pyrifoliaNakaiandXinjiangPear, meanwhilethe3
P.caleryanacultivarsdidntclustertogether, probablybe-
causeXinjiangPearismoreleantoBretschneideriRehdand
P.pyrifoliaNakaiingenecomposition.TheresearchbyZhao
Guofang[ 8] suggestedthatXinjiangPearhadaclosergenetic
relationshipwithP.BretschneideriRehdandP.communisL,
andmaybewasthehybridcultivarofP.BretschneideriRehd
andP.communisL, whichisconsistentwiththeresearchre-
sultsofZouLeminbypalynology[ 9].Theexperimentalre-
sultssuggestedthatthesmalesteuclideandistancebetween
Xinjiang Pear and P.pyrifolia Nakai, P.communisL,
P.BretschneideriRehdaswelasP.ussuriensisMaximwere
7.746, 7.746, 7.810 and8.165 , respectively.Thecluster
resultssuggestedthatwhenthresholdvaluewas15, Xinjiang
PearcultivarsLanzhouchangbaandHuachangbaclusteredto-
getherfirstly, andthenclusteredtogetherwithP.communisL
cultivars.Comprehensiveresearchessuggestthatthereisa
closergeneticrelationshipbetweenXinjiangPearandP.com-
munisL.
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幼苗期小麦热激蛋白的诱导研究
宋松＊　(南京农业大学生命科学学院 ,江苏南京 210095)
摘要　高温是小麦产量提高的一个重要限制因素 ,小麦受到高温胁迫时会产生一系列热激蛋白来适应这种逆境 ,不同发育阶段的热激蛋白
存在差异 ,不同抗热作物品种中热激蛋白也存在差异 ,因此 ,研究抗热性不同的小麦品种中热激蛋白差异对小麦耐热性的研究具有重要的理
论意义与实践意义。该实验利用SDS-PAGE电泳技术对 2种不同的小麦品系在 34.37、40℃下处理 1 ～ 7 h产生的热激蛋白进行研究 , 发现
其可溶性蛋白电泳图谱与常温下的有区别 ,其中新蛋白的出现说明高温能够诱导新蛋白。 71321-9中发现的 3种新蛋白的分子量分别为 97.
4kD、49kD、37kD,另外还发现 3种蛋白质含量明显升高 ,这 3种蛋白质的分子量范围在 17～ 66kD。 71321-4小麦中发现了 2种新增热激蛋
白(37kD和 29kD)及 1种蛋白质的含量明显增加。这些蛋白可能与小麦幼苗抗热性有关 ,其含量的增加能够提高幼苗的抗热性。
关键词　热激蛋白;小麦;品种;电泳
作者简介　宋松(1989-),男,江苏淮安人 ,本科生 ,专业:生命科学。 ＊通讯作者。
收稿日期　2009-09-07　　修回日期　2009-09-29
(上接第 4页)
利用 AFLP分子标记鉴定新疆梨的分类地位(摘要)
鲁凤娟＊　(中国环境管理干部学院 ,河北秦皇岛 066004)
[目的]对新疆梨的分类地位提供 DNA水平上的依据。
[方法]以健康植株新梢上的成熟叶片为材料 ,采用CTAB法提取DNA。利用 AFLP分子标记技术 ,通过统计软件SPSSforWindows计算供试
材料之间的欧氏距离 ,利用离差平方和法进行聚类分析 ,输出欧氏距离表并产生聚类树状图 ,对新疆梨进行分类研究。
[结果]当阈值为 15时 ,新疆梨品种兰州长把和花长把首先聚在一起 ,然后再与西洋梨品种巴梨、红巴梨、红茄 、茄梨 、保利阿斯卡、朱丽比恩
和新疆梨品种其力阿木提聚在一起。 7个新疆梨品种中 ,群体内品种之间欧式距离为 2.646 ～ 10.050,与砂梨系统 、西洋梨系统、白梨系统、秋
子梨系统各品种间最小欧式距离分别为 7.746、7.746、7.810、8.165。
[结论]新疆梨与西洋梨系统有较近亲缘关系。
关键词　分子标记;新疆梨;分类地位;AFLP
基金项目　河北省自然科学基金(302240)。
作者简介　鲁凤娟(1978-),女 ,河北昌黎人 ,讲师 ,从事园艺与绿色食品研究。 ＊通讯作者。
收稿日期　2009-12-04　　修回日期　2010-01-12
30 AgriculturalScience＆TechnologyVol.11, No.1, 2010