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Infection and Migration of Marked Rhizobia in Alfalfa (Medicago sativa) Bud Seedlings under the Action of Exogenous Substances

几种外源物质对内生根瘤菌侵染苜蓿芽苗并在植株体内运移的影响



全 文 :!23" !6#
 Vol.23  No.6
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犱狅犻:10.11733/j.issn.10070435.2015.06.019
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‘’“ Authorforcorrespondence,Email:shishl@gsau.edu.cn,A!ò’“¤¡y
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ן犚.犿犲犾犻犾狅狋犻GNfe犛.犿犲犾犻犾狅狋犻12531f(^ C:Y{)l{ӝË656.83%e303.19%,ýü犛.12531f”wŸl
{Óð¾h-l3.64%。IAA¾@üÖÕן犚.GNfeS.12531{ӝË18.34e12.11Ÿ,üw=犛.12531
Ë163.09%,¢犚.GNfÎs¾h-l75.45%。IAA¾@eÎsÖÕܳŸ犚.GNf׺•h-l4.13%,Èɕ
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:S154.381    [\]^_:A     [`aZ:10070435(2015)06125906
犐狀犳犲犮狋犻狅狀犪狀犱犕犻犵狉犪狋犻狅狀狅犳犕犪狉犽犲犱犚犺犻狕狅犫犻犪犻狀犃犾犳犪犾犳犪(犕犲犱犻犮犪犵狅狊犪狋犻狏犪)
犅狌犱犛犲犲犱犾犻狀犵狊狌狀犱犲狉狋犺犲犃犮狋犻狅狀狅犳犈狓狅犵犲狀狅狌狊犛狌犫狊狋犪狀犮犲狊
LIJianfeng1,2,ZHANGShuqing1,2,SHIShangli2,MIAOYangyang2,HUOPinghui2
(1.KeyLaboratoryofGuizhouBioresourceDevelopmentandUtilization,InstituteofsoieandEnvironmentBioremediation
inkarsthabitats,GuizhouNormalColege,Guiyang,GuizhouProvince550025,China;2.KeyLaboratoryofGrasslandEcosystemof
MinistryofEducation,SinoU.S.CentersforGrazingLandEcosystemSustainability,GansuAgriculturalUniversity,Lanzhou,Gansu
Province730070,China)
犃犫狊狋狉犪犮狋:Endogenous犚犺犻狕狅犫犻犪existedwidelyinthewholebodyandseedsofleguminousplants,butthe
specificmigrationalpathway,migrationalpatternandthefactorsthatinfluencingendogenous犚犺犻狕狅犫犻犪mi
grationwerenotclear.3indoleaceticacid(IAA),lanthanumchloride(LaCl3)andextracelularpolysac
charidesfromother犚犺犻狕狅犫犻犪wereusedinthisstudytoconductrootimmersiononthebudseedlingsof
犕犲犱犻犮犪犵狅狊犪狋犻狏犪cv.‘Gannong5’,inordertocomparethemigrationaldynamicandquantitativedistribu
tionofthemarked犚犺犻狕狅犫犻犪containingCFP(Cyanfluorescentprotein)undertheactionofthe3exogenous
substances,andtofurtherexploretheinfluenceofendogenousrhizobiastrain(isolatedfromhostplant)
andexogenousstrain(purchasedfromstrainconservationcenter,Chineseacademyofsciences)犚犺犻狕狅犫犻犪.
Theresultsshowedthat犚犺犻狕狅犫犻狌犿犿犲犾犻犾狅狋犻.GNf(endogenousmarkedstrain)celswere11.43and1.71
timesofCKinrootandstemundertheactionofextracelularpolysaccharides,respectively.TheLaCl3of
50mg·L-1increasedthenumberof犚.犿犲犾犻犾狅狋犻.GNfand犛.犿犲犾犻犾狅狋犻.12531f(exogenousmarkedstrain)in
therootby656.83%and303.19%,anddecreasedthenumberof犛.12531finstemtothe3.64%ofCK.
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IAAincreased犚.GNfandS.12531celsintheseedlingrootby18.34and12.11times,respectively,
whileforthenumberinstem,犛.12531hadincreasedto163.09%,while犚.GNfdecreasedto75.45%.
IAAcouldsignificantlydecrease犚.GNfcelsincotyledontothe4.13%ofCK,indicatingthatalthe3
exogenoussubstancesincreasedthecelsofmarked犚犺犻狕狅犫犻犪inroot.Thequantitydifferenceofmarked
rhizobiacelnumbersindifferentpartsofbudseedlingsunderexogenoussubstancefreeconditionsindica
tedthatthemigrationalpathwayof犚犺犻狕狅犫犻狌犿amongrootstemcotyledoninthebudseedlingshadthese
lectivebarriersofonlyalowingendogenous犚犺犻狕狅犫犻犪topass.
犓犲狔狑狅狉犱狊:Endogenousrhizobia;犕犲犱犻犮犪犵狅狊犪狋犻狏犪.L;Migrationaldynamic;3IAA;Exopolysaccharides
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GNfג{{B[8],qA{B犛犻狀狅狉犺犻狕狅犫犻狌犿犿犲犾犻犾狅
狋犻12531s=<-ÇGGoåp=nÂÃ,犚犺犻狕狅犫犻
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eß­©¢HoaglandHëeß­[11]。eß­˜
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˜—gßZeeß­¸”121℃45â{26min。
1.2.2 …$Y[b†zz‡„eˆz‰Š‹Œ‡
(EPS)]  Ž   { k ­ ˜ ¬:c 4℃ å ` l
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ŸC{~°Mê{Bb^&°
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\¢ewŒƒñѾŒx

1.2.4 zD‰Š‹Œ‡×؅$Y[z‘
’“”xx]•–?@ c犛.12531f{k­A犚.
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ü25℃ñtgß。24h>uSÖÕ,¢y{Œ¿ 
5q>,cžxo¾Ô×、Ôw、ܳ、z³4 x。
cFxo6Œx2õQ>úHy{•˜Ï=

¢ý
þȉâ{3min,y{Œ¿ 8q>,úHy{D
ÿ=

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{onÀ
,4000rpmon5min,u—²­0.2mL
âùHTY¼ÓgßZ,24h>M¬{¸G&KL。
˜ðË犛.12531fl¾@¾h-,3¾@34QY。
犚.GNfl¾@ˆò¤—,b^&°±¢犛.
12531f{Bb^&°ly{Âu­。
1.2.5 IAA×؅$Y[z‘’“”xx]•
–?@ c0.08mg·L-1IAAˀµ¯:Y{l
{k­=

žÒ¾@ˆò¤|„

1.2.6 LaCl3 ×؅$Y[z‘’“”xx]
•–?@ c50mg·L-1LaCl3 ˀµ¯:Y{
l{k­=

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يlíâ¦Ð
[13]。
£¤{Bxˆlb^&°
=¢H‘XŽ2l|„Æb£M¤

§°T=^
C{犛.12531f~°ŸC{犚.GNflb^&°>
yJUVÖÕ
,24h>eB:Y{¸*”z³=[
S

Ì ÖÕ w = 犛.12531fl × º d h - ^ S
589.02%,aê!" (犘<0.05)(û 1)。ן
犛.12531f{Ó×dh-^S26.64%,¢eJyJ
犛.12531fl¾@Í,ܳŸ¸ØÙ:Y{Sa。I
òÆÇseAb^&°=l#ê~|„xÜhÙ
ŠŽlíâ~XnlLúUÐÙ=
[9]。
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³ŸC{gß­ª×¾@=

ŸC{”ÙŠŸl×
º!"^H^C{

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Xn

Ì^C{ш‹‰hIòö~óô

W1 Ržö(ižö)ŽRA-*ˆižö(Ržö)ö./"lm24犺,]ñ"$ö˜%&ÈÃßi@‚?X»
Fig.1 Quantitativedistributionoffluorescentmarkedrhizobiainthetissuesofbudseedlingsafterbeingimmersedinthe
mixtureoftheextracelularpolysaccharidesofexogenousbacteria(endogenousbacteria)andendogenousbacterial
(exogenousbacterial)solutionfor24h
Ô

F¾@z³Ÿ¸*[S:Y{

ͤ
Note:Nofluorescentproteinmarkedstrainswerefoundedintheeuphylasofalseedlings,thesameasbelow
  ŸCג{犚.GNf”~°Mê{Bb^&°
>
,犚.GNf”ÖÕ×ewŸl:Y{×x2¾h
-l11.43e1.71Ÿ,aê!"(犘<0.05)。ÖÕ
ܳŸ犚.GNflC{×ð¾h-l61%。F¾@
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z³Ÿ¸*[S:Y{

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C{犛.12531f/Ÿ
C{犚.GNf)€ÙŠŽÖÕ×ewle9ÙF
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ŸC{b^&°£eq^C{÷±»
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2.2 =+(3犐犃犃)/]ñ"$ö˜%&6i(
)@AB
ŽÃwÙËӟò“ìl’¢
[14]。
sdÐlIAAö~»ýbŽ„üýbŽst~
Ë

Ë(bŽldÞÐ
[3],
üŽíWIJ=ë
h€){lbŽÎy†C~Îs

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W2 0.08犿犵·犔-13犐犃犃*ˆö./"lm24犺,]ñ"$ö˜%&i@X»
Fig.2 Thedistributionofmarkedrhizobiainbudseedlingsafterbeingimmersedin
themixtureof0.08mg·L-13IAAandbacterialsolutionfor24h
  sû2su,”˜:Y{{­hÖժ׾@
öU=

¤ÞvË^C3IAA£eém:Yג
{”ÖÕӟlx¹ Ì

¢eém:Y{”F 
Ìl׺

:Y{~°IAAª×¾@>,ÖÕן
l犚.GNfe犛.12531fl{Ó׺k]ò*,x2
oh-Ë+18.34e12.11Ÿ,aê!"(犘<
0.05);¢”wŸŸC{{Ó׺Îs¾h-l
75.45%,^ C{ѝË+163.09%。eJ:Y{
¸*”ÖÕlz³Ÿ[S

ŸC{犚.GNf”ܳl
{Ó׺`ð¾h-l4.13%。IseðsH
IAAŠ‹y!’¢H×I,ü×ÈlýbŽùÇ
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× Ÿ&=ŽlíâÓI™æt¬

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b\ýbùÇAŽ°5Gl|×¼¦{ŠÙx
ˆIAAle9。+y™¡`ÄSf|ג{{B
”³gßvwÍçxˆG&w

”ÙŠÓŸ`ç
öX#xˆ\Rs{ÓAيùl‘XŽ2LÅÙ
ŠŽxˆqÂ^يӟlÃwŒã
[16]。
^C
G&w‡çŽÃŽýbŽfg(ºÆ„es&
„

I1òs’¾ßxÂ̟Gý{

¾ý{r
GHŽÂþ™leß&
[4]。
2.3 犔犪3+÷¨/]ñ"$ö˜%&6i()@A
B
ÇG)€ŽÓŸÞ

{Óȉl"&„1
òè£eWŽýbŽ2¥çLúيŽlí
âXnÓI
。Ca2+’¾ŽÓŠ‹l‘üoÜ,”
‘XVÅ=Q=¨’¢
[18]。La3+’¾Ca2+”ýb
=løù#¨X
[19],
sà²Ca2+/Bql‘
X‚Å

émŽh)€ÇGlíâXn

¤Þ
La3+`çAb7bŽlP&„¥7˜°,Þ7ý
bŽÞõ
[1]。
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W3 50犿犵·犔-1犔犪犆犾3 *ˆö./"24犺,]ñ"$ö˜%&i@X»
Fig.3 Thedistributionofmarkedrhizobiainbudseedlingsafterbeingimmersed
inthemixtureof50mg·L-1LaCl3andbacterialsolutionfor24h
  sû3su,50mg·L-1lLaCl3 süן
犚.GNfe犛.12531fl{Ó׺x2oh-Ë
656.83%e303.19%,aê!"(犘<0.05)。¢
La3+t¬+ŸC{犚.GNfÊܳewl“ì。Ì
h^C:Y{犛.12531f,La3+l`”!"Îs+{
ӔwŸl׺

üñð¾h-l3.64%。ÈÉ
La3+~»+:Y{ÊUVÖÕ× l)”e“ì,
¢£ŽH:Y{ÊweܳŸl“ì

òˆ‰s
eðsHÖÕl× ”LaCl3 l’¢Í,×Iȉ
lbŽùǝ(

ýCa2+W#¨>Ca2+ABq
ùl‘XVÅúGàš
[18],
يŽh€){Ó
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