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Isolation, Identification and Diversity Analysis of Fungal Endophyte in Astragalus strictus

茎直黄芪内生真菌分离鉴定及其多样性分析



全 文 :!23" !6#
 Vol.23  No.6
! " # $
ACTA AGRESTIA SINICA
   2015$  11%
  Nov.  2015
犱狅犻:10.11733/j.issn.10070435.2015.06.018
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:Q933331    [\]^_:A     [`aZ:10070435(2015)06125207
犐狊狅犾犪狋犻狅狀,犐犱犲狀狋犻犳犻犮犪狋犻狅狀犪狀犱犇犻狏犲狉狊犻狋狔犃狀犪犾狔狊犻狊狅犳
犉狌狀犵犪犾犈狀犱狅狆犺狔狋犲犻狀犃狊狋狉犪犵犪犾狌狊狊狋狉犻犮狋狌狊
CAODandan,LUHao,LIXiaomeng,GUOYazhou,XUERuixu,
WANGHuan,QUANHaiyun,ZHAOBaoyu
(ColegeofVeterinaryMedicine,NorthwestA&FUniversity,Yangling,ShaanxiProvince712100,China)
犃犫狊狋狉犪犮狋:Fungalendophytewereisolatedfrom犃狊狋狉犪犵犪犾狌狊狊狋狉犻犮狋狌狊samplescolectedfromGongbujiangda
cityofTibettoinvestigatethespeciesdiversityanddistributionofendophyte.Thespeciesandgenusof
endophytewereidentifiedbythetraditionalmorphologyandrDNAITSsequenceanalysis,anddiversity
indexwascalculatedbyBiodap.Theresultsshowedthat336fungalcolonieswereisolatedfrom424sam
ples,whichwerecombinedto21kindsofdifferentendophyticfungus.Underaltheconditionsofsteriliza
tion,犃犾狋犲狉狀犪狉犻犪狋犲狀狌犻狊狊犻犿犪 wasthedominantspeciesinthesespecieswiththerelativefrequencyof
32.14%,folowedwith犝狀犱犻犳犻犾狌犿狅狓狔狋狉狅狆犻狊(19.05%).Diversityindexesshowedthatthediversityin
dexoffungalendophytedifferedfromdifferenttissues.TheShannonWeinerindex,simpsonindexand
evennessindexoffungalendophyteinflowerwerealhighest,whichwas2.160.154,and0.80,respec
tively,folowedbyleafandstem.Inconclusion,therewasanobviousdifferenceinquantity,speciesand
diversityoftheendophyteamongdifferenttissuesof犃狊狋狉犪犵犪犾狌狊狊狋狉犻犮狋狌狊.Flowerwasthemostvulnerable
tocolonizationbytheendophyte,whosediversitywasthemaximum.
犓犲狔狑狅狉犱狊:犃狊狋狉犪犵犪犾狌狊狊狋狉犻犮狋狌狊;Fungalendophyte;Isolationandidentification;Speciesdiversity
CDEF
:20140918;GHEF:20141210
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Table1 Isolationresultsofendophyticfungifromdifferenttissuesof犃.狊狋狉犻犮狋狌狊
6Œ Ì
Tissues
â{Þù
Sterilizationtime
75%0
75%ethanol/s
2%NaClO/min
6Œ©×
Tissuesnumber
{¸¹×
Colonynumber
xo²
Isolationrate/%
¹xo²
Totalisolation
rate/%
Ó
Flower
30 2.5 47 45 95.74
30 3.0 50 47 94.00
40 3.5 46 31 67.39
86.01
³
Leaf
30 2.5 48 46 95.83
30 3.0 50 49 98.00
40 3.5 43 19 44.19
80.85
w
Stem
30 2.5 48 42 87.50
30 3.0 46 36 78.26
40 3.5 46 21 45.65
70.71
2.2 i=õöø
5xoS„ŸGz{336B,R¥H)Goz
‰°¾67B¥H。sû1su,JH1Y37A
犆犾犪犱狅狊狆狅狉犻狌犿犮犾犪犱狅狊狆狅狉犻狅犻犱犲狊,JH2J21 A 犕犪犾
犫狉犪狀犮犺犲犪犮犻狀狀犪犿狅犿犲犪 l M Ý ~ ¸ ¾ 100%,
JH1Y34A 犃犾狋犲狉狀犪狉犻犪犱犪狋狌狉犻犮狅犾犪,JH1Y410 A
犝狀犱犻犳犻犾狌犿 狅狓狔狋狉狅狆犻狊,JH2H22 A 犝犾狅犮犾犪犱犻狌犿
犿狌犾狋犻犳狅狉犿犲,JH1Y310 A JH2H31,JH2H32,
犃犾狋犲狉狀犪狉犻犪sp.,犃犾狋犲狉狀犪狉犻犪狋犲狀狌犻狊狊犻犿犪,犘犺狅犿狅狆狊犻狊
sp.,JH1Y45 A JH3H16,犉狌狊犪狉犻狌犿 犳犾狅犮犮犻犳犲
狉狌犿,犉狌狊犪狉犻狌犿狋狅狉狌犾狅狊狌犿 lMÝ~¾99%,Èɞ
ŠÞ=IdO«

_sGožJ!

žÒŸGz{×
YžFXIJú±ýÆ°{¸¥H

È2),sc
{B»¼J!Go

ÆÇÈÉ
,336BŸGz{x
!H7ÿ、9†、10<、13!、21J,ž=18[Q•
oJ
,3[*•oJ。wxÁJŸGz{”F6Œ
 Ìlx¹KL¶xo²0û2F6。
2.3 òóœôi=õö¦ùÃ]9X»­
”FÙxoælŸGz{=
,犇狅狋犺犻犱犲狅犿狔犮犲狋犲狊
sp.,犛狋犪犵狅狀狅狊狆狅狉狅狆狊犻狊犪狊狋狉犪犵犪犾犻,犛.犮狌犮狌狉犫犻狋犪
犮犲犪狉狌犿,犃犾狋犲狉狀犪狉犻犪狋犲狀狌犻狊狊犻犿犪,犝狀犱犻犳犻犾狌犿狅狓狔狋狉狅
狆犻狊e犘犲狔狉狅狀犲犾犾犪犲犪狆犻狀狅犱犲犾犾犪”wxÁJlw、³
eÓ=¸Wxoæ

sóITŸGz{y6Œ#ê
~
。犘犺狅犿狅狆狊犻狊sp.e犔犲狆狋狅狊狆犺犪犲狉狌犾犻狀犪狊犪犮犮犺犪狉犻
犮狅犾犪ð”³=xoæ,犉狌狊犪狉犻狌犿狋狅狉狌犾狅狊狌犿 e犝犾狅
犮犾犪犱犻狌犿 犿狌犾狋犻犳狅狉犿犲 ð”Ó=xoæ,Ì 犇犻犱狔
犿犲犾犾犪犳犪犫犪犲ð”w=xoæ,ÈÉIñJŸGz{
loŽÿÙòol6Œ#ê~

û2)。”FÙxo
ælŸGz{=犃.狋犲狀狌犻狊狊犻犿犪xo²(32.14%)ß
^

ý”w

³eÓ=xo²¸d^

¾wxÁJŸ
Gz{=lzÃ{!

ž4¾犝.狅狓狔狋狉狅狆犻狊。
4521
!6# ț›¡:wxÁJŸGz{xoGo¶ž|}~x£
v2 òóœôi=õö@ˆR,-Ò­®
Table2 Mainmorphologicalcharacteristicsofenophyteisolatedfrom犃.狊狋狉犻犮狋狌狊
{B[õ
Species
xo Ì
Tissues
¥H)#$ Morphologicalcharacteristics
{¸v‰¥H
Obversemorphology
{¸ô‰¥H
Backmorphology
G&±Ð
Growthspeed/
mm·d-1
JH3J45 wStem
{‰M

×ØG&

{¸dÞ

ã

=nÚCM7ÞCM
,^
ȐM

ë!
@M

ë! 0.8
JH1J13
³Leaf
ÓFlower
{‰M

×ØG&

{¸dÞ

ã

z#¾M

=#=ù`´M".
M

d#=ùä´M".ªWM

u­!
z#¾M

=#=ù`
´M".M

d#=ù
ä´M".ªWM


!
4.17
JH1Y310 ³Leaf {‰M、×ØG&,{¸òO、ÅS、M~,M,ë! M,ë! 0.1
JH1H11
wStem
³Leaf
ÓFlower
{‰M

×ØG&

{¸òO

ã

ÅS

{¸=ùÁM".CM CM

ë! 1.17
JH3J17
wStem
³Leaf
ÓFlower
{‰M

×ØG&

{¸dÞ

".ã=ùåQ

=ù@M".ª
M

u­!
=ù@M".ÁM

7
ÞÁM

u­!
2.75
JH3J12
wStem
³Leaf
ÓFlower
{‰M

×ØG&

{¸dÞ

ã

@M

ë!
=ùCM

7ސM

ë
!
6.3
JH2H31
wStem
ÓFlower
{‰@M

iGG&

{¸dÞ

ÅS

=ù@M

7ÞAÁM

u­! CM

u­! 7.56
JH2H32
wStem
ÓFlower
{‰CM

iGG&

{¸dÞ

ÅS

7ށã

=ù@M

".CM CM

u­! 3.89
JH1Y410
wStem
³Leaf
ÓFlower
{‰M

iGG&

{¸dÞ

ÅS

Ú@M

ë!
=ùÚ@M

7ސM

ë!
0.25
JH2H22 ÓFlower {‰M、×ØG&,{¸dÞ、=ùåQ".ã,Ÿ CM7ސM,ë! CM,ë! 3.33
JH1Y37
³Leaf
ÓFlower
{‰AÁM

×ØG&

{¸dÞ

=ùåQ".ã

ë!
=ùÁ".ûÉ

7ސ
M

ë!

JH1Y45
³Leaf
ÓFlower
{‰M

iGG&

{¸dÞ

M

ÅS

ë! M

ë! 3.16
JH3H16 ÓFlower {‰AÁM、iGG&,{¸dÞ、ÅS,¦MÁMMù,ë!
¦MÁMMù

7Þ¦
M

ë!
8.33
JH1Y210 ³Leaf {‰AÁM、×ØG&,{¸*O、=ùåQ".ã,ë!
CM

7ÞAÁMM

ë
!
1.44
JH1Y112
³Leaf
ÓFlower
{‰AÁM

×ØG&

{¸*O

ÅS

ȉæå£ã

ë! CM

ë! 1.33
JH1Y211 ³Leaf {‰M、iGG&,{¸dÞ、ȉæå£ã,M,7ÞÆçM,ë! M,ë! 1.33
JH3H12
ÓFlower
wStem
{‰M

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W1 Iú犐犜犛ûüýþÿfÔÕ@Ç«!Ê"
Fig.1 PhylogenetictreeconstructedwiththeprogramNeighbor-joining(NJ)basedonITSsequences
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Fig.2 Thepopulationcompositionofendophytefrom犃.狊狋狉犻犮狋狌狊
2.4 òóœôi=õö>¦ABÉX„
0È3F6,”M¤g߶xovwÍwxÁ
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Table3 Thediversityindexesoftheendophyticfungiindifferenttissuesof犃.狊狋狉犻犮狋狌狊
Ä×Index
Ä׸3
Indexabbreviation
ÓFlower ³Leaf wStem ¹ÓTotal
Ž4vbÄ×
ShannonWeinerIndex
犎 2.16 1.92 1.78 2.12
c:;Ä×
Simpsondiversityindex
犇 0.154 0.248 0.223 0.195
1/犇 6.494 4.039 4.477 5.128
¸ËÐÄ×
Evennessindex
犈 0.80 0.69 0.77 0.68
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