Pathogen Identification and Screening of Fungicides Against Microsorum pteropus Anthracnose Disease

doi:10.11924/j.issn.1000-6850.casb2022-0613

Abstract

Abstract:

In order to clarify the pathogen causing Microsorum pteropus anthracnose disease in Gaoyao district, Zhaoqing City, Guangdong Province, and to screen better effect fungicides against the disease indoor, the pathogen was obtained by using the tissue separation method. The representative isolates were verified according to Koch’s rule. Combined with morphological characteristics and phylogenetic tree based on ITS, ACT, CAL and CHS-1 genes, the pathogen was identified, and the indoor toxicity of six fungicides (42.4% pyraclostrobin·metiram WG, 75% trifloxystrobin·tebuconazole WG, 35% 400 g/L fluopyram·tebuconazole SC, 325 g/L difenoconazole· azoxystrobin SC, 60% pyraclostrobin·fluxapyroxad WG, 10% difenoconazole WG) was determined by mycelia growth rate method. Colletotrichum fructicola was the pathogen causing anthracnose disease on Microsorum pteropus. The tested six fungicides had certain inhibitory effect on the mycelia growth of C. fructicola. Among them, 10% difenoconazole WG had the strongest inhibitory activity with moderate inhibitory concentration (EC₅₀) of 0.1083 μg/mL. Followed by 325 g/L difenoconazole· azoxystrobin SC and 75% trifloxystrobin· tebuconazole WG, the above three fungicides can be recommended as field control agents.

Key words: Guangdong Province, Microsorum pteropus, Colletotrichum fructicola, Koch’s rule, the indoor toxicity

CUI Yiping, SONG Xiaobing, YAN Jiechun, HUANG Feng, LING Jinfeng, PENG Aitian. Pathogen Identification and Screening of Fungicides Against Microsorum pteropus Anthracnose Disease[J]. Chinese Agricultural Science Bulletin, 2023, 39(24): 120-126.

Figures/Tables 6

References 24

[1]	刘丽萍, 高洁, 李玉. 植物炭疽菌属Colletotrichum真菌研究进展[J]. 菌物研究, 2020, 18(4):266-281.
[2]	LIU L P, ZHANG L, WANG Y, et al. Distribution, identification and characterization of Colletotrichum lineola and C. panacicola causing anthracnose on ginseng in northeast China[J]. Crop protection, 2020a, 205265.
[3]	LIU L P, ZHANG L, QIU P L, et al. Leaf spot of polygonatum odoratum caused by Colletotrichum spaethianum[J]. Journal of general plant pathology, 2020b, 86:157-161. doi: 10.1007/s10327-019-00903-4
[4]	DEAN R, KAN J A L V, PRETORIUS Z A, et al. The top 10 fungal pathogens in molecular plant pathology[J]. Molecular plant pathology, 2012, 13(4):414-430. doi: 10.1111/j.1364-3703.2011.00783.x pmid: 22471698
[5]	韩淑清. 观赏水草的种类调查及栽培技术的研究[D]. 杭州: 浙江大学, 2005.
[6]	薛春丽, 王卫江, 高永红. 铁皇冠孢子人工培养研究[J]. 云南农业科技, 2008(5):16-17.
[7]	薛春丽, 郭俊明, 王卫疆, 等. 铁皇冠微室培养及性器官发育研究[J]. 安徽农业科学, 2009, 37(12):5737-5738.
[8]	WANG J, WANG Y, XU F. Restoration of environment, preferably water and wetland ecosystem polluted by cadmium involves growing polypodiaceae plant in environment polluted by cadmium[J]. Peking university, 2009.
[9]	LAN X Y, YAN Y Y, YANG B, et al. Differential expression of proteins in the leaves and roots of cadmium-stressed Microsorum pteropus, a novel potential aquatic cadmium hyperaccumulator[J]. Science of the total environment, 2018, 642:1369e1377.
[10]	LAN X Y, YANG B, YAN Y Y, et al. Resistance mechanisms and their difference between the root and leaf of microsorum pteropus-a novel potential aquatic cadmium hyperaccumulator[J]. Science of the total environment, 2018, 616e617,480e490.
[11]	LAN X Y, YAN Y Y, YANG B, et al. Subcellular distribution of cadmium in a novel potential aquatic hyperaccumulator-microsorum pteropus[J]. Environmental pollution, 2019, 248:1020e1027.
[12]	何艳林, 余建波, 许宝红. 观赏水草栽培管理及病害防治[J]. 休闲渔业, 2006, 5:21-22.
[13]	GARDES M, BRUNS T D. ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts[J]. Molecular ecology, 1993, 2(2):113-118. doi: 10.1111/j.1365-294X.1993.tb00005.x URL
[14]	WHITE T J, BRUNS T, LEE S, et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[A]. INNIS M A, GELFAND D H, SNINSKY J J. PCR protocols: a guide to methods and applications[M]. San Diego: Academic Press, 1990:315-322.
[15]	CARBONE I, KOHN L M. A method for designing primer sets for speciation studies in filamentous ascomycetes[J]. Mycologia, 1999, 91(3):553-556. doi: 10.1080/00275514.1999.12061051 URL
[16]	O'DONNELL K, NIRENBERG H I, AOKI T, et al. A multigene phylogeny of the Gibberella fujikuroi species complex:detection of additional phylogenetically distinct species[J]. Mycoscience, 2000, 41(1):61-78. doi: 10.1007/BF02464387 URL
[17]	JAYAWARDENA R S, HYDE K D, DAMM U, et al. Notes on currently accepted species of Colletotrichum[J]. Mycosphere, 2016, 7(8):1192-1260. doi: 10.5943/mycosphere URL
[18]	李佳星, 袁静, 周沙, 等. 325 g/L苯甲·嘧菌酯悬浮剂在油茶中的残留及消解动态[J]. 农药, 2022, 61(2):123-126.
[19]	DAMM U, CANNON P F, WOUDENBERG J H C, et al. The Colletotrichum boninense species complex[J]. Studies in mycology, 2012, 73:1-36. doi: 10.3114/sim0002 URL
[20]	覃丽萍, 苏琴, 谢玲, 等. 杧果炭疽病菌Colletotrichum asianum和C. fructicola的危害特征及对杀菌剂的敏感性初探[J]. 植物保护, 2020, 46(1):197-202.
[21]	LIN S R, YU S Y, CHANG T D, et al. First report of anthracnose caused by Colletotrichum fructicola on tea in Taiwan[J]. Plant disease, 2021, 105:3.
[22]	LEE D M, HASSAN O, KIM C H. First report of anthracnose of peach (Prunus persica) caused by Colletotrichum fructicola in Korea[J]. Plant disease, 2020, 104:5. doi: 10.1094/PDIS-104-01-0005 URL
[23]	CHEN X G, LIU C L, LIU J A, et al. First report of colletotrichum fructicola causing anthracnose on Camellia yuhsienensis in China[J]. Plant disease, 2022, 106:1.
[24]	HOFER K M, BRAITHWAITE M, BRAITHWAITE L J, et al. First report of Colletotrichum fructicola, C. perseae, and C. siamense causing anthracnose disease of avocado (Persea americana) in New Zealand[J]. Plant disease, 2021, 105:5. doi: 10.1094/PDIS-105-01-0005 URL

供试杀菌剂	别名	生产产家
42.4%唑醚·氟酰胺 SC 21.2%吡唑醚菌酯 21.2%氟唑菌酰胺	施乐健	巴斯夫（中国）有限公司
75%肟菌戊唑醇 WG 25%肟菌酯 50%戊唑醇	拿敌稳	拜耳（中国）有限公司
35% 400g/L氟菌·戊唑醇 SC 17.5%氟吡菌酰胺 17.5%戊唑醇	露娜润	拜耳（中国）有限公司
325g/L 苯甲.嘧菌酯 SC 125g/L苯醚甲环唑 200g/L嘧菌酯	阿米妙收/凯典	先正达集团公司
60%唑醚.代森联 WG 5%吡唑醚菌酯 55%代森联	百泰	巴斯夫（中国）有限公司
10%苯醚甲环唑 WDG	恶醚唑	先正达集团公司

供试杀菌剂	别名	生产产家
42.4%唑醚·氟酰胺 SC 21.2%吡唑醚菌酯 21.2%氟唑菌酰胺	施乐健	巴斯夫（中国）有限公司
75%肟菌戊唑醇 WG 25%肟菌酯 50%戊唑醇	拿敌稳	拜耳（中国）有限公司
35% 400g/L氟菌·戊唑醇 SC 17.5%氟吡菌酰胺 17.5%戊唑醇	露娜润	拜耳（中国）有限公司
325g/L 苯甲.嘧菌酯 SC 125g/L苯醚甲环唑 200g/L嘧菌酯	阿米妙收/凯典	先正达集团公司
60%唑醚.代森联 WG 5%吡唑醚菌酯 55%代森联	百泰	巴斯夫（中国）有限公司
10%苯醚甲环唑 WDG	恶醚唑	先正达集团公司

种		菌株编号	Genbank登录号
种		菌株编号	ACT	CAL		CHS1
果生炭疽菌C. fructicola		THG1-1	ON016174	ON016173		ON016175
果生炭疽菌C. fructicola		ZH6	MT500908	MT500952		MT500996
果生炭疽菌C. fructicola		Y2	MZ724635	MZ724640		MZ724650
果生炭疽菌C. fructicola		YM7	MT500907	MT500951		MT500995
果生炭疽菌C. fructicola		PLY-6-1	MT980992	MW046766		MW032332
果生炭疽菌C. fructicola		YSG-4-2	MT981003	MW046777		MW032343
隐秘炭疽菌C. aenigma		ZH2	MT500880	MT500924		MT500968
隐秘炭疽菌C. aenigma		YM8	MT500879	MT500923		MT500967
暹罗炭疽菌C. siamense	YH-YX-6		MW513863		MW513911	MW513887
暹罗炭疽菌C. siamense	ZS-HX-85		MW513855		MW513903	MW513879
暹罗炭疽菌C. siamense	LFY-8		OL830177		OL830174	OL830171
茶树炭疽病菌C. camelliae	HXQT-3		MN525809		MN525825	MN525846
茶树炭疽病菌C. camelliae	YN23		MN525815		MN525834	MN525855
哈锐炭疽菌C. horii	SD011		MN070079		MN070084	MN070093
哈锐炭疽菌C. horii	GX014		MN070078		MN070088	MN070097
亚洲炭疽菌C. asianum	MC38		MN626847		MN605542	MN626900
亚洲炭疽菌C. asianum	MC6		MN626846		MN605539	MN626891

种		菌株编号	Genbank登录号
种		菌株编号	ACT	CAL		CHS1
果生炭疽菌C. fructicola		THG1-1	ON016174	ON016173		ON016175
果生炭疽菌C. fructicola		ZH6	MT500908	MT500952		MT500996
果生炭疽菌C. fructicola		Y2	MZ724635	MZ724640		MZ724650
果生炭疽菌C. fructicola		YM7	MT500907	MT500951		MT500995
果生炭疽菌C. fructicola		PLY-6-1	MT980992	MW046766		MW032332
果生炭疽菌C. fructicola		YSG-4-2	MT981003	MW046777		MW032343
隐秘炭疽菌C. aenigma		ZH2	MT500880	MT500924		MT500968
隐秘炭疽菌C. aenigma		YM8	MT500879	MT500923		MT500967
暹罗炭疽菌C. siamense	YH-YX-6		MW513863		MW513911	MW513887
暹罗炭疽菌C. siamense	ZS-HX-85		MW513855		MW513903	MW513879
暹罗炭疽菌C. siamense	LFY-8		OL830177		OL830174	OL830171
茶树炭疽病菌C. camelliae	HXQT-3		MN525809		MN525825	MN525846
茶树炭疽病菌C. camelliae	YN23		MN525815		MN525834	MN525855
哈锐炭疽菌C. horii	SD011		MN070079		MN070084	MN070093
哈锐炭疽菌C. horii	GX014		MN070078		MN070088	MN070097
亚洲炭疽菌C. asianum	MC38		MN626847		MN605542	MN626900
亚洲炭疽菌C. asianum	MC6		MN626846		MN605539	MN626891

供试杀菌剂	杀菌剂浓度/(μg/mL)		抑制率/%	供试杀菌剂	杀菌剂浓度/(μg/mL)		抑制率/%
75%肟菌·戊唑醇 WG	0.1		39.51	60%唑醚.代森联SC	0.1		19.69
	0.5		56.63		0.5		31.24
	2		61.34		2		43.08
	10		64.76		10		68.47
	50		71.75		50		77.75
35% 400g/L氟菌·戊唑醇SC		0.1	2.43	42.4%唑醚·氟酰胺 WG		0.1	24.69
		0.5	36.8			0.5	40.99
		2	53.07			2	58.07
		10	56.06			10	62.27
		50	74.75			50	62.27
325g/L苯甲·嘧菌酯 SC 325 g/L		0.1	40.23	10%苯醚甲环唑 WG		0.2	58.54
		0.5	54.78			0.4	72.83
		2	81.88			0.6	75.78
		10	83.31			0.8	76.71
		50	85.02			1	82.61
CK		-	-