Specific PCR Detection of Causal Agent of Persimmon Anthracnose Colletotrichum horii

doi:10.11924/j.issn.1000-6850.casb2026-0050

Abstract

Abstract:

Persimmon anthracnose is one of the most serious fungal diseases in persimmon production. Colletotrichum horii is the dominant pathogen. Early rapid detection is very important for disease prevention and control. To develop a rapid and specific PCR method for detecting Colletotrichum horii and apply it to field monitoring and rapid diagnosis of persimmon anthracnose, this study used multiple Colletotrichum species as test materials, compared the gene sequences of TUB2 (β-tubulin) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase), designed four primer pairs, and screened out two specific primer pairs (THF1/THR2, GHF1/GHR2) for PCR amplification, sensitivity testing, and artificial inoculation experiments. The results showed that the primer pair THF1/THR2 based on the TUB2 gene amplified a specific 211 bp band, and the primer pair GHF1/GHR2 based on the GAPDH gene amplified a specific 171 bp band; both primer pairs achieved a detection sensitivity of 100 pg/μL, and in artificially inoculated persimmon leaves, both primer pairs consistently detected the corresponding target bands. In conclusion, the PCR detection method established in this study is highly specific and sensitive, providing reliable technical support for field monitoring and rapid diagnosis of persimmon anthracnose.

Key words: persimmon, anthracnose, Colletotrichum horii, PCR detection, TUB2, GAPDH, primer design

CLC Number:

S436.63

CHEN Hailing, LI Qili, ZHOU Hao, SONG Jinkang, HUANG Suiping, CHEN Xiaolin, ZHANG Yu, TANG Lihua. Specific PCR Detection of Causal Agent of Persimmon Anthracnose Colletotrichum horii[J]. Chinese Agricultural Science Bulletin, 2026, 42(11): 140-150.

Figures/Tables 10

References 26

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No.	Strain no.	Species	Host	Accession no.
1	FJ11-1	Colletotrichum asianum	Mangifera indica	MH636523^a
2	GD10-1	Colletotrichum siamense	Mangifera indica	MH636415^a
3	HN47-2	Colletotrichum fructicola	Mangifera indica	MH636458^a
4	HN23-5	Colletotrichum cordylinicola	Mangifera indica	MH636464^a
5	HN32-1	Colletotrichum tropicale	Mangifera indica	MH636466^a
6	HN42-2	Colletotrichum gigasporum	Mangifera indica	MH636467^a
7	JH7-2	Colletotrichum plurivorum	Mangifera indica	MW507147^a
8	YN31-4	Colletotrichum cliviicola	Mangifera indica	MH636508^a
9	YN32-2	Colletotrichum endophytica	Mangifera indica	MH636509^a
10	YN33-1	Colletotrichum liaoningense	Mangifera indica	MH636504^a
11	YN51-1	Colletotrichum scovillei	Mangifera indica	MH636512^a
12	GZ15-1	Colletotrichum gloeosporioides	Mangifera indica	MH636440^a
13	GZ23-3	Colletotrichum musae	Mangifera indica	MH636444^a
14	YN34-1	Colletotrichum karstii	Mangifera indica	MH636506^a
15	PLG3-1	Colletotrichum horii	Persimmon	MT982860^a
16	YB53-2	Epicoccum sorghinum	Mangifera indica	/
17	EGY16A-1	Pseudofusicoccum violaceum	Violet	/
18	YB43-2	Alternaria brassicae	Mangifera indica	/
19	HF	Neoscytalidium dimidiatum	Hylocereus undulatus	N
20	TY6-1	Phyllosticta capitalensis	Mangifera indica	PV259393^a
21	TY9-2	Botryosphaeria	Vitis vinifera	/
22	YB33-1	Exserohilum	Mangifera indica	/
23	YMWK	Rhizoctonia solani	Zea mays	N
24	FY27-1	Phanerochaete sordida	/	/
25	HZ40-3	Neopestalotiopsis sonneratae	Prunus salicina	N
26	TEY15-1	Fusarium sulawesiense	Prunus salicina	OM432482^a
27	STJ	Penicillium digitatum	Citrusreticulata	N
28	G2-1	Pantoea anthophila	Mangifera indica	/
29	Y5-1	Pantoea vagans	Mangifera indica	OL413424.1^b
30	G10-1	Pantoea cypripedii	Mangifera indica	/
31	Y11-2	Pantoea dispersa	Mangifera indica	/
32	Y12-1	Xanthomonas	Mangifera indica	/
33	PLY8-1	Colletotrichum fructicola	Persimmon	MT982874^a
34	PLY4-2	Colletotrichum karstii	Persimmon	MT982871^a
35	YSY2-2	Colletotrichum cliviae	Persimmon	MT982887^a
36	PLY16-2	Colletotrichum siamense	Persimmon	MT982879^a
37	GCG7-2	Colletotrichum horii	Persimmon	MT982820^a
38	GCG-6-1	Colletotrichum horii	Persimmon	MT982818^a
39	PLG-5-1A	Colletotrichum horii	Persimmon	MT982862^a

No.	Strain no.	Species	Host	Accession no.
1	FJ11-1	Colletotrichum asianum	Mangifera indica	MH636523^a
2	GD10-1	Colletotrichum siamense	Mangifera indica	MH636415^a
3	HN47-2	Colletotrichum fructicola	Mangifera indica	MH636458^a
4	HN23-5	Colletotrichum cordylinicola	Mangifera indica	MH636464^a
5	HN32-1	Colletotrichum tropicale	Mangifera indica	MH636466^a
6	HN42-2	Colletotrichum gigasporum	Mangifera indica	MH636467^a
7	JH7-2	Colletotrichum plurivorum	Mangifera indica	MW507147^a
8	YN31-4	Colletotrichum cliviicola	Mangifera indica	MH636508^a
9	YN32-2	Colletotrichum endophytica	Mangifera indica	MH636509^a
10	YN33-1	Colletotrichum liaoningense	Mangifera indica	MH636504^a
11	YN51-1	Colletotrichum scovillei	Mangifera indica	MH636512^a
12	GZ15-1	Colletotrichum gloeosporioides	Mangifera indica	MH636440^a
13	GZ23-3	Colletotrichum musae	Mangifera indica	MH636444^a
14	YN34-1	Colletotrichum karstii	Mangifera indica	MH636506^a
15	PLG3-1	Colletotrichum horii	Persimmon	MT982860^a
16	YB53-2	Epicoccum sorghinum	Mangifera indica	/
17	EGY16A-1	Pseudofusicoccum violaceum	Violet	/
18	YB43-2	Alternaria brassicae	Mangifera indica	/
19	HF	Neoscytalidium dimidiatum	Hylocereus undulatus	N
20	TY6-1	Phyllosticta capitalensis	Mangifera indica	PV259393^a
21	TY9-2	Botryosphaeria	Vitis vinifera	/
22	YB33-1	Exserohilum	Mangifera indica	/
23	YMWK	Rhizoctonia solani	Zea mays	N
24	FY27-1	Phanerochaete sordida	/	/
25	HZ40-3	Neopestalotiopsis sonneratae	Prunus salicina	N
26	TEY15-1	Fusarium sulawesiense	Prunus salicina	OM432482^a
27	STJ	Penicillium digitatum	Citrusreticulata	N
28	G2-1	Pantoea anthophila	Mangifera indica	/
29	Y5-1	Pantoea vagans	Mangifera indica	OL413424.1^b
30	G10-1	Pantoea cypripedii	Mangifera indica	/
31	Y11-2	Pantoea dispersa	Mangifera indica	/
32	Y12-1	Xanthomonas	Mangifera indica	/
33	PLY8-1	Colletotrichum fructicola	Persimmon	MT982874^a
34	PLY4-2	Colletotrichum karstii	Persimmon	MT982871^a
35	YSY2-2	Colletotrichum cliviae	Persimmon	MT982887^a
36	PLY16-2	Colletotrichum siamense	Persimmon	MT982879^a
37	GCG7-2	Colletotrichum horii	Persimmon	MT982820^a
38	GCG-6-1	Colletotrichum horii	Persimmon	MT982818^a
39	PLG-5-1A	Colletotrichum horii	Persimmon	MT982862^a

物种名称	菌株名称	寄主	地理来源	GenBank登录号
物种名称	菌株名称	寄主	地理来源	ITS	TUB2	GAPDH
C. cliviae	GCY1-2A	Persimmon	China	MT982835	MW046811	MW032377
C. cliviae	GCY4-2	Persimmon	China	MT982837	MW046813	MW032379
C. cliviae	PLY14-2	Persimmon	China	MT982878	MW046854	MW032420.1
C. cliviae	YSY2-2	Persimmon	China	MT982887	MW046863	MW032429
C. fructicola	PLY8-1	Persimmon	China	MT982874	MW046850	MW032416
C. fructicola	YSY1-2	Persimmon	China	MT982886	MW046862	MW032428
C. fructicola	YSY3-3	Persimmon	China	MT982888	MW046864	MW032430
C. fructicola	YSY5-2	Persimmon	China	MT982889	MW046865	MW032431
C. fructicola	YSY8-3	Persimmon	China	MT982890	MW046866	MW032432
C. horii	PLG5-1A	Persimmon	Pingle, Guilin	MT982862	MW046838	MW032404
C. horii	PLG3-1	Persimmon	Pingle, Guilin	MT982860	MW046836	MW032402
C. horii	PLG3-2	Persimmon	China	MT982861	MW046837	MW032403
C. horii	PLG6-1	Persimmon	China	MT982864	MW046840	MW032406
C. karstii	LPY4-2B	Persimmon	China	MT982858	MW046834	MW032400
C. karstii	PLY7-2	Persimmon	China	MT982873	MW046849	MW032415
C. karstii	GCY21-3	Persimmon	China	MT982850	MW046826	MW032392
C. karstii	PLY2-2	Persimmon	China	MT982869	MW046845	MW032411
C. karstii	PLY4-2	Persimmon	China	MT982871	MW046847	MW032413
C. siamense	GCG17-1	Persimmon	China	MT982829	MW046805	MW032371
C. siamense	PLY16-2	Persimmon	China	MT982879	MW046855	MW032421
C.asianum	FJ11-1	Mangifera indica	China	MH636523	MH622733	MH681402
C.asianum	FJ18-3	Mangifera indica	China	MH636528	MH622738	MH681407
C.asianum	FJ36-6	Mangifera indica	China	MH636539	MH622749	MH681418
C.asianum	QZ-3	Mangifera indica	China	MF039834	MF039805	MF040765
C.asianum	YN55-1	Mangifera indica	China	MH636503	MH622713	MH681382
C.cliviicola	YN8-2	Mangifera indica	China	MH636487	MH622697	MH681366
C.cliviicola	YN31-4	Mangifera indica	China	MH636508	MH622718	MH681387
C.cordylinicola	HN23-5	Mangifera indica	China	MH636464	MH622674	MH681343
C.endophyticum	YN32-2	Mangifera indica	China	MH636509	MH622719	MH681388
C.endophyticum	HN37-2	Mangifera indica	China	MH636453	MH622663	MH681331
C.endophyticum	HN37-6	Mangifera indica	China	MH636453	MH622663	MH681332
C.endophyticum	YN32-6	Mangifera indica	China	MH681390	MH636511	MH622721
C.fructicola	GZ15-1	Mangifera indica	China	MH636440	MH622650	MH681319
C.fructicola	HN47-2	Mangifera indica	China	MH636458	MH622668	MH681337
C.fructicola	FJ32-6	Mangifera indica	China	MH636534	MH622744	MH681413
C.fructicola	FJ35-5	Mangifera indica	China	MH636538	MH622748	MH681417
C.fructicola	GZ1-2	Mangifera indica	China	MH636427	MH622637	MH681306
C.gigasporum	HN42-2	Mangifera indica	China	MH636467	MH622677	MH681346
C.gloeosporioides	GZ14-G-1	Mangifera indica	China	MH636439	MH622649	MH681318
C.liaoningense	YN33-1	Mangifera indica	China	MH636504	MH622714	MH681383
C.musae	GZ23-3	Mangifera indica	China	MH636444	MH622654	MH681323
C.plurivorum	JH7-2	Mangifera indica	China	JF898133	MW507372	MW507371
C.scovillei	HN44-3	Mangifera indica	China	MH636457	MH622667	MH681336
C.scovillei	YN51-1	Mangifera indica	China	MH636512	MH622722	MH681391
C.scovillei	HN44-2	Mangifera indica	China	MH636456	MH622666	MH681335
C.siamense	YN40-1-2	Mangifera indica	China	MH636499	MH622709	MH681378
C.siamense	GD8-4	Mangifera indica	China	MH636416	MH622626	MH681293
C.siamense	GD10-1	Mangifera indica	China	MH636415	MH622625	MH681294
C.siamense	YN56-1-1	Mangifera indica	China	MH636497	MH622707	MH681376
C.siamense	YN56-2-1	Mangifera indica	China	MH636498	MH622708	MH681377
C.tropicale	HN32-1	Mangifera indica	China	MH636466	MH622676	MH681345

物种名称	菌株名称	寄主	地理来源	GenBank登录号
物种名称	菌株名称	寄主	地理来源	ITS	TUB2	GAPDH
C. cliviae	GCY1-2A	Persimmon	China	MT982835	MW046811	MW032377
C. cliviae	GCY4-2	Persimmon	China	MT982837	MW046813	MW032379
C. cliviae	PLY14-2	Persimmon	China	MT982878	MW046854	MW032420.1
C. cliviae	YSY2-2	Persimmon	China	MT982887	MW046863	MW032429
C. fructicola	PLY8-1	Persimmon	China	MT982874	MW046850	MW032416
C. fructicola	YSY1-2	Persimmon	China	MT982886	MW046862	MW032428
C. fructicola	YSY3-3	Persimmon	China	MT982888	MW046864	MW032430
C. fructicola	YSY5-2	Persimmon	China	MT982889	MW046865	MW032431
C. fructicola	YSY8-3	Persimmon	China	MT982890	MW046866	MW032432
C. horii	PLG5-1A	Persimmon	Pingle, Guilin	MT982862	MW046838	MW032404
C. horii	PLG3-1	Persimmon	Pingle, Guilin	MT982860	MW046836	MW032402
C. horii	PLG3-2	Persimmon	China	MT982861	MW046837	MW032403
C. horii	PLG6-1	Persimmon	China	MT982864	MW046840	MW032406
C. karstii	LPY4-2B	Persimmon	China	MT982858	MW046834	MW032400
C. karstii	PLY7-2	Persimmon	China	MT982873	MW046849	MW032415
C. karstii	GCY21-3	Persimmon	China	MT982850	MW046826	MW032392
C. karstii	PLY2-2	Persimmon	China	MT982869	MW046845	MW032411
C. karstii	PLY4-2	Persimmon	China	MT982871	MW046847	MW032413
C. siamense	GCG17-1	Persimmon	China	MT982829	MW046805	MW032371
C. siamense	PLY16-2	Persimmon	China	MT982879	MW046855	MW032421
C.asianum	FJ11-1	Mangifera indica	China	MH636523	MH622733	MH681402
C.asianum	FJ18-3	Mangifera indica	China	MH636528	MH622738	MH681407
C.asianum	FJ36-6	Mangifera indica	China	MH636539	MH622749	MH681418
C.asianum	QZ-3	Mangifera indica	China	MF039834	MF039805	MF040765
C.asianum	YN55-1	Mangifera indica	China	MH636503	MH622713	MH681382
C.cliviicola	YN8-2	Mangifera indica	China	MH636487	MH622697	MH681366
C.cliviicola	YN31-4	Mangifera indica	China	MH636508	MH622718	MH681387
C.cordylinicola	HN23-5	Mangifera indica	China	MH636464	MH622674	MH681343
C.endophyticum	YN32-2	Mangifera indica	China	MH636509	MH622719	MH681388
C.endophyticum	HN37-2	Mangifera indica	China	MH636453	MH622663	MH681331
C.endophyticum	HN37-6	Mangifera indica	China	MH636453	MH622663	MH681332
C.endophyticum	YN32-6	Mangifera indica	China	MH681390	MH636511	MH622721
C.fructicola	GZ15-1	Mangifera indica	China	MH636440	MH622650	MH681319
C.fructicola	HN47-2	Mangifera indica	China	MH636458	MH622668	MH681337
C.fructicola	FJ32-6	Mangifera indica	China	MH636534	MH622744	MH681413
C.fructicola	FJ35-5	Mangifera indica	China	MH636538	MH622748	MH681417
C.fructicola	GZ1-2	Mangifera indica	China	MH636427	MH622637	MH681306
C.gigasporum	HN42-2	Mangifera indica	China	MH636467	MH622677	MH681346
C.gloeosporioides	GZ14-G-1	Mangifera indica	China	MH636439	MH622649	MH681318
C.liaoningense	YN33-1	Mangifera indica	China	MH636504	MH622714	MH681383
C.musae	GZ23-3	Mangifera indica	China	MH636444	MH622654	MH681323
C.plurivorum	JH7-2	Mangifera indica	China	JF898133	MW507372	MW507371
C.scovillei	HN44-3	Mangifera indica	China	MH636457	MH622667	MH681336
C.scovillei	YN51-1	Mangifera indica	China	MH636512	MH622722	MH681391
C.scovillei	HN44-2	Mangifera indica	China	MH636456	MH622666	MH681335
C.siamense	YN40-1-2	Mangifera indica	China	MH636499	MH622709	MH681378
C.siamense	GD8-4	Mangifera indica	China	MH636416	MH622626	MH681293
C.siamense	GD10-1	Mangifera indica	China	MH636415	MH622625	MH681294
C.siamense	YN56-1-1	Mangifera indica	China	MH636497	MH622707	MH681376
C.siamense	YN56-2-1	Mangifera indica	China	MH636498	MH622708	MH681377
C.tropicale	HN32-1	Mangifera indica	China	MH636466	MH622676	MH681345

设计区	引物		序列(5'-3')
TUB2	上游引物	THF1	GGGGCTAACCAGAGTTCTTTCA
	下游引物	THR2	TTAGCGGTCAACCACCAATGT
	上游引物	THF3	GTTACATTTGCAAGAACAACGG
GAPDH	上游引物	GHF1	CCTCCAAACTTGTCATGACTTCACTTC
	下游引物	GHR2	TGAATGTGACAGACGGGGCTTATT
	上游引物	GHF3	TTATCCGCACCCCGCCG