Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (1): 129-140.doi: 10.16420/j.issn.0513-353x.2020-1033
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YU Lin1,2, SHE Xiaoman1, TANG Yafei1, LAN Guobing1, LI Zhenggang1, HE Zifu1,2,*()
Received:
2021-07-18
Revised:
2021-08-19
Online:
2022-01-25
Published:
2022-01-24
Contact:
HE Zifu
E-mail:hezf@gdppri.com
CLC Number:
YU Lin, SHE Xiaoman, TANG Yafei, LAN Guobing, LI Zhenggang, HE Zifu. Identification of Pathogen Causing Allium ascalonicum Anthracnose in Guangdong[J]. Acta Horticulturae Sinica, 2022, 49(1): 129-140.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2020-1033
目的序列 Target sequence | 引物名称 Primer | 引物序列(5′-3′) Primer sequence | 参考文献 Reference |
---|---|---|---|
ITS | ITS1F | CTTGGTCATTTAGAGGAAGTAA | Gardes & Bruns, |
ITS4 | TCCTCCGCTTATTGATATGC | White et al., | |
ACT | ACT-512F | ATGTGCAAGGCCGGTTTCGC | Carbone & Kohn, |
ACT-783R | TACGAGTCCTTCTGGCCCAT | Carbone & Kohn, | |
CHS1 | CHS-79F | TGGGGCAAGGATGCTTGGAAGAAG | Carbone & Kohn, |
CHS-354R | TGGAAGAACCATCTGTGAGAGTTG | Carbone & Kohn, | |
GAPDH | GDF1 | GCCGTCAACGACCCCTTCATT | Guerber et al., |
GDR1 | GGGTGGAGTCGTACTTGAGCATGT | Guerber et al., | |
TUB2 | T1 | AACATGCGTGAGATTGTAAGT | O’ Donnell & Cigelnik, |
Btub4R | CCRGAYTGRCCRAARACRAAGTTGTC | Aveskamp et al., |
Table 1 PCR Primers for ITS,ACT,CHS1,GAPDH and TUB2 genes amplification
目的序列 Target sequence | 引物名称 Primer | 引物序列(5′-3′) Primer sequence | 参考文献 Reference |
---|---|---|---|
ITS | ITS1F | CTTGGTCATTTAGAGGAAGTAA | Gardes & Bruns, |
ITS4 | TCCTCCGCTTATTGATATGC | White et al., | |
ACT | ACT-512F | ATGTGCAAGGCCGGTTTCGC | Carbone & Kohn, |
ACT-783R | TACGAGTCCTTCTGGCCCAT | Carbone & Kohn, | |
CHS1 | CHS-79F | TGGGGCAAGGATGCTTGGAAGAAG | Carbone & Kohn, |
CHS-354R | TGGAAGAACCATCTGTGAGAGTTG | Carbone & Kohn, | |
GAPDH | GDF1 | GCCGTCAACGACCCCTTCATT | Guerber et al., |
GDR1 | GGGTGGAGTCGTACTTGAGCATGT | Guerber et al., | |
TUB2 | T1 | AACATGCGTGAGATTGTAAGT | O’ Donnell & Cigelnik, |
Btub4R | CCRGAYTGRCCRAARACRAAGTTGTC | Aveskamp et al., |
种类 | 菌株 | GenBank登录号GenBank accession number | ||||
---|---|---|---|---|---|---|
Species | Isolate | ITS | ACT | CHS1 | GAPDH | TUB2 |
Colletotrichum lilii | CBS 186.30 | GU227811 | GU227909 | GU228301 | GU228203 | GU228105 |
CBS 109214 | GU227810 | GU227908 | GU228300 | GU228202 | GU228104 | |
C. liriopes | CBS 119444 | GU227804 | GU227902 | GU228294 | GU228196 | GU228098 |
C. spaethianum | BLC1 | MW295524 | MW298835 | MW298836 | MW298837 | MW298838 |
CBS 167.49 | GU227807 | GU227905 | GU228297 | GU228199 | GU228101 | |
CBS 100063 | GU227808 | GU227906 | GU228298 | GU228200 | GU228102 | |
CBS 101631 | GU227809 | GU227907 | GU228299 | GU228201 | GU228103 | |
WYC1 | MW295525 | MW298839 | MW298840 | MW298841 | MW298842 | |
C. tofieldiae | CBS 495.85 | GU227801 | GU227899 | GU228291 | GU228193 | GU228095 |
C. trichellum | CBS 217.64 | GU227812 | GU227910 | GU228302 | GU228204 | GU228106 |
C. verruculosum | IMI 45525 | GU227806 | GU227904 | GU228296 | GU228198 | GU228100 |
C. gloeosporioides | CBS 112999 | JX010152 | JX009531 | JX009818 | JX010056 | JX010445 |
C. theobromicola | ICMP 18649 | JX010294 | JX009444 | JX009869 | JX010006 | JX010447 |
C. truncatum | CBS 151.35 | GU227862 | GU227960 | GU228352 | GU228254 | GU228156 |
Monilochaetes infuscans | CBS 869.96 | JQ005780 | JQ005843 | JQ005801 | JX546612 | JQ005864 |
Table 2 Reference isolates used in this study and their GenBank accession numbers
种类 | 菌株 | GenBank登录号GenBank accession number | ||||
---|---|---|---|---|---|---|
Species | Isolate | ITS | ACT | CHS1 | GAPDH | TUB2 |
Colletotrichum lilii | CBS 186.30 | GU227811 | GU227909 | GU228301 | GU228203 | GU228105 |
CBS 109214 | GU227810 | GU227908 | GU228300 | GU228202 | GU228104 | |
C. liriopes | CBS 119444 | GU227804 | GU227902 | GU228294 | GU228196 | GU228098 |
C. spaethianum | BLC1 | MW295524 | MW298835 | MW298836 | MW298837 | MW298838 |
CBS 167.49 | GU227807 | GU227905 | GU228297 | GU228199 | GU228101 | |
CBS 100063 | GU227808 | GU227906 | GU228298 | GU228200 | GU228102 | |
CBS 101631 | GU227809 | GU227907 | GU228299 | GU228201 | GU228103 | |
WYC1 | MW295525 | MW298839 | MW298840 | MW298841 | MW298842 | |
C. tofieldiae | CBS 495.85 | GU227801 | GU227899 | GU228291 | GU228193 | GU228095 |
C. trichellum | CBS 217.64 | GU227812 | GU227910 | GU228302 | GU228204 | GU228106 |
C. verruculosum | IMI 45525 | GU227806 | GU227904 | GU228296 | GU228198 | GU228100 |
C. gloeosporioides | CBS 112999 | JX010152 | JX009531 | JX009818 | JX010056 | JX010445 |
C. theobromicola | ICMP 18649 | JX010294 | JX009444 | JX009869 | JX010006 | JX010447 |
C. truncatum | CBS 151.35 | GU227862 | GU227960 | GU228352 | GU228254 | GU228156 |
Monilochaetes infuscans | CBS 869.96 | JQ005780 | JQ005843 | JQ005801 | JX546612 | JQ005864 |
Fig. 5 Microstructure characteristics of the pathogen of Allium ascalonicum anthracnose A,B:Conidiophores with conidia on the top;C:Conidia;D:Appressoria;E,F:Seta;G:Acervuli on the leaf of A. ascalonicum.
Fig. 6 Phylogenetic tree based on sequences of ITS,ACT,CHS1,GAPDH and TUB2 genes Values at the node of each clade are parsimony bootstrap above 50%,● indicates test strains.
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