一种来源于葡萄叶片的真菌(Quambalaria cyanescens)的鉴定及培养特性

doi:10.6048/j.issn.1001-4330.2022.10.024

摘要/Abstract

摘要：

【目的】研究一株来源于新疆吐鲁番市葡萄病叶上真菌的种类及分类地位。【方法】观察菌株的培养性状,测量初生分生孢子和次生分生孢子的大小,基于ITS、LSU基因片段采用最大简约法、最大似然法和贝叶斯法构建系统发育树;将该纯化菌株活化培养至不同培养基种类、光照条件、pH值和培养温度等4个因素的不同水平下培养,混合正交试验,分析最适生长条件。【结果】培养基种类、光照条件、pH值和培养温度等4个因素的交互作用对菌株Pt4-1菌落生长具有显著影响,菌株Pt4-1在PDA、SDA培养基上较CMA培养基生长更快,菌株在 SDA培养基、半光照、pH5、温度25 ℃条件下的生长速率最大,平均生长速率为2.29 cm/d;菌株Pt4-1在CMA培养基、半光照、pH 5、温度25 ℃条件下的生长速率最小,平均生长速率为0.55 cm/d。【结论】菌株Pt4-1为蓝色刺孢霉Quambalaria cyanescens;菌株Pt4-1在不同培养条件下菌落形态不同,在PDA、SDA培养基上生长良好,可分泌紫色色素,在CMA培养基上生长但不分泌色素;随着培养温度的升高,色素分泌减少。

关键词: 蓝色刺孢霉Quambalaria cyanescens; 葡萄叶片; 种类鉴定; 培养特征

Abstract:

【Objective】 To clarify the species and taxonomic status of a rare fungus from grapevine diseased leaves in Turpan region of Xinjiang.【Methods】 Culture traits were observed in the strains and the size of primary and secondary conidia were measured.Based on ITS and LSU sequences, phylogenetic tree was constructed by Maximum likelihood, the maximum parsimony method and Bayesian method.Meanwhile, the purified strain was cultured to different levels of four factors, such as different medium types, light conditions, pH and culture temperature, and combined orthogonal experiment were conducted to explore the optimum growth conditions.【Results】 The results showed that the interaction of four factors, including medium species, light conditions, pH values and culture temperature, was found to have a significant effect on strain Pt4-1and the strain grew faster on PDA, SDA medium than on CMA medium.The growth rate of strain Pt4-1 grew the fastest under SDA medium, semi-light, pH 5 and temperature 25 ℃, with the average growth rate of 2.29 cm/d.The growth rate of strain Pt4-1 was the weakest under CMA medium, half-light, pH 5 and temperature 25 ℃, the average growth rate was 0.55 cm/d.【Conclusion】 After comprehensive analysis, Pt4-1 was identified as Quambalaria cyanescens.Mixed orthogonal tests indicated that the growth rate of strain Pt4-1 was faster on PDA and SDA media than that on CMA medium and could secret purple pigment, but no purple pigment on CMA medium.With the increase of culture temperature, the secretion of pigment decreased.

Key words: Quambalaria cyanescens; grape leaf; species identification; cultivation characteristics

中图分类号:

S188

翟亚伟, 赵颖, 李胜男, 朱妙菲, 潘雯, 巴音达拉, 马荣. 一种来源于葡萄叶片的真菌(Quambalaria cyanescens)的鉴定及培养特性[J]. 新疆农业科学, 2022, 59(10): 2546-2553.

ZHAI Yawei, ZHAO Ying, LI Shengnan, ZHU Miaofei, PAN Wen, Bayandala , MA Rong. Identification of a Rare Fungus (Quambalaria cyanescens) Derived from Grape Leaves[J]. Xinjiang Agricultural Sciences, 2022, 59(10): 2546-2553.

图/表 4

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种名 Taxon	菌株编号 Strain	寄主 Host	分布地区 Location	采集人 Collector	基因片段GenBank号
种名 Taxon	菌株编号 Strain	寄主 Host	分布地区 Location	采集人 Collector	ITS	LSU
Q.fabacearum	URM 7756	Mimosa tenuiflora	Brazil	J.D.P.Bezerra	MG253664	MG253665
Q.pusilla	MUCC 828		Australia	Burgess,T.I.	MN959759
Q.rugosae	CBS 145601	Eucalyptus rugosa	Australia: SA	Crous,P.W.	MN162014	MN162212
Q.tasmaniae	CBS 145602	Eucalyptus sp.	Australia: SA	Crous,P.W.	MN162015	MN162213
	CPC:25462	Eucalyptus sp.	Australia: TAS	Crous,P.W.	MN162016
Q.eucalypti	CBS 118844	Eucalyptus grandis	South Africa	M.J.Wingfield	NR111214	NG042417
	CMW11678	Eucalyptus grandis	South Africa	L Lombard	DQ317626	DQ317619
Q.coyrecup	WAC12948	Corymbia calophylla	Western Australia	T.Paap	DQ823433	DQ823446
Q.pitereka	WAC12956	Corymbia ficifolia	Western Australia	T.Paap	DQ823428	DQ823435
	DAR 19773	Corymbia eximia	Australia:New South Wales	Zhou,X.	NR111242	NG060071
Q.simpsonii	CBS 124772	Eucalyptus tintinnans	Edith Falls Australia	BA Summerell	GQ303290	GQ303321
	CBS 124773	Eucalyptus sp.	Lamphoon, Thailand	R Cheewangkoon	GQ303291	GQ303322
Q.cyanescens	CBS 357.73	skin of man	Netherlands	TF Visser	DQ317622	DQ317615
	CBS 876.73	Eucalyptus pauciflflora	New South Wales,Australia	VF Brown	DQ317623	DQ317616
	WAC 12954	Corymbia calophylla	Western Australia	T Paap	DQ823420	DQ823442
	BRIP 48396	Corymbia citriodora	Beaudesert, Queens land, Australia	GS Pegg	EF444874
	11PU348	Implants	China	Fan,X.	KF953496	KF953497
	MK 742	Pistacia vera	Turkey	Kolarik,M.	AM261920
	MK 808	Olea europaea	Syria:Krak des Chevaliers	Kolarik,M.	AM261921
	MK 1710	Quercus sp.	Bulgaria	Kolarik,M.	AM261922
	MK 617	Carpinus betulus	Hungary	Kolarik,M.	AM261923
	MK 1617	Olea europea	Spain:Andalusia, Huelva province, Galaroza	Kolarik,M.	AM261924
	WAC 12953	Corymbia ficifolia	Australia	Paap,T.	DQ823422	DQ823443
	CCF 4580	Prunus sp.	USA	Kolarik,M.	HG421953
	QY 229	red kojic rice	China	Zhang,Z.G.	HM013823
	AUMC 6293	air of citrus plantation	Egypt	Zeinab Soliman	JQ425376
	AUMC 6294	citrus juice	Egypt	Zeinab Soliman	JQ425382
	IRAN 2465C	Punica granatum	Iran	M.E.Vahedi Darmiyan	KX377510
	CBS 127352	Betula pendula	Russia	AB Antropova	HG799002	MH875972
	CCTU 1684	Vitis vinifera	Iran	A Narmani	MN006031
	UM 1095	skin of man	Malaysia:Kuala Lumpur	Ng,K.P.	KT186106	KT186107
	Pt 4-1	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019644	MW019675
	Pt 4-2	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019645	MW019676
	Pt 6-1	Vitis vinifera	Turpan, a city in Xinjiang Province, China	Ma,R	MW019646	MW019677
	Pt 6-2	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019647	MW019678
Microstroma juglandis	RB 2042	Juglans regia	Germany	Bereng.Sacc.	DQ317634	DQ317617

种名 Taxon	菌株编号 Strain	寄主 Host	分布地区 Location	采集人 Collector	基因片段GenBank号
种名 Taxon	菌株编号 Strain	寄主 Host	分布地区 Location	采集人 Collector	ITS	LSU
Q.fabacearum	URM 7756	Mimosa tenuiflora	Brazil	J.D.P.Bezerra	MG253664	MG253665
Q.pusilla	MUCC 828		Australia	Burgess,T.I.	MN959759
Q.rugosae	CBS 145601	Eucalyptus rugosa	Australia: SA	Crous,P.W.	MN162014	MN162212
Q.tasmaniae	CBS 145602	Eucalyptus sp.	Australia: SA	Crous,P.W.	MN162015	MN162213
	CPC:25462	Eucalyptus sp.	Australia: TAS	Crous,P.W.	MN162016
Q.eucalypti	CBS 118844	Eucalyptus grandis	South Africa	M.J.Wingfield	NR111214	NG042417
	CMW11678	Eucalyptus grandis	South Africa	L Lombard	DQ317626	DQ317619
Q.coyrecup	WAC12948	Corymbia calophylla	Western Australia	T.Paap	DQ823433	DQ823446
Q.pitereka	WAC12956	Corymbia ficifolia	Western Australia	T.Paap	DQ823428	DQ823435
	DAR 19773	Corymbia eximia	Australia:New South Wales	Zhou,X.	NR111242	NG060071
Q.simpsonii	CBS 124772	Eucalyptus tintinnans	Edith Falls Australia	BA Summerell	GQ303290	GQ303321
	CBS 124773	Eucalyptus sp.	Lamphoon, Thailand	R Cheewangkoon	GQ303291	GQ303322
Q.cyanescens	CBS 357.73	skin of man	Netherlands	TF Visser	DQ317622	DQ317615
	CBS 876.73	Eucalyptus pauciflflora	New South Wales,Australia	VF Brown	DQ317623	DQ317616
	WAC 12954	Corymbia calophylla	Western Australia	T Paap	DQ823420	DQ823442
	BRIP 48396	Corymbia citriodora	Beaudesert, Queens land, Australia	GS Pegg	EF444874
	11PU348	Implants	China	Fan,X.	KF953496	KF953497
	MK 742	Pistacia vera	Turkey	Kolarik,M.	AM261920
	MK 808	Olea europaea	Syria:Krak des Chevaliers	Kolarik,M.	AM261921
	MK 1710	Quercus sp.	Bulgaria	Kolarik,M.	AM261922
	MK 617	Carpinus betulus	Hungary	Kolarik,M.	AM261923
	MK 1617	Olea europea	Spain:Andalusia, Huelva province, Galaroza	Kolarik,M.	AM261924
	WAC 12953	Corymbia ficifolia	Australia	Paap,T.	DQ823422	DQ823443
	CCF 4580	Prunus sp.	USA	Kolarik,M.	HG421953
	QY 229	red kojic rice	China	Zhang,Z.G.	HM013823
	AUMC 6293	air of citrus plantation	Egypt	Zeinab Soliman	JQ425376
	AUMC 6294	citrus juice	Egypt	Zeinab Soliman	JQ425382
	IRAN 2465C	Punica granatum	Iran	M.E.Vahedi Darmiyan	KX377510
	CBS 127352	Betula pendula	Russia	AB Antropova	HG799002	MH875972
	CCTU 1684	Vitis vinifera	Iran	A Narmani	MN006031
	UM 1095	skin of man	Malaysia:Kuala Lumpur	Ng,K.P.	KT186106	KT186107
	Pt 4-1	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019644	MW019675
	Pt 4-2	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019645	MW019676
	Pt 6-1	Vitis vinifera	Turpan, a city in Xinjiang Province, China	Ma,R	MW019646	MW019677
	Pt 6-2	Vitis vinifera	Turpan City, Xinjiang Uygur Autonomous Region	Ma,R	MW019647	MW019678
Microstroma juglandis	RB 2042	Juglans regia	Germany	Bereng.Sacc.	DQ317634	DQ317617