草莓属植物种质资源对炭疽病抗性的离体评价

doi:10.3864/j.issn.0578-1752.2019.20.009

摘要/Abstract

摘要：

【目的】 对草莓属植物12个草莓野生种24份材料、1个栽培种41个品种、种间杂交种6个品种共71份种质资源的炭疽病抗性进行评价,为草莓属植物种质资源的利用提供依据。【方法】 以从草莓上分离得到的果生炭疽菌（Colletotrichum fructicola）为接种病原,采用离体接种法将炭疽菌孢子悬浮液（1×10 ⁶个分生孢子/mL）均匀接种在草莓叶片的叶面、叶柄上。置于28℃条件下保湿培养4 d,然后分别对每个叶片的发病情况进行调查,统计叶面病斑数量、叶面最大病斑直径、叶柄最大病斑长度。运用IBM SPSS 15.0软件对不同供试材料的叶面病斑数量、叶面病斑直径、叶柄病斑长度进行相关性分析。根据叶柄病斑长度对每个叶片的病害严重程度进行分级,计算每份材料的病情指数,以接种炭疽菌后叶柄的病情指数为依据对供试材料的炭疽病抗性进行评价。以草莓种类为固定因素,供试材料为随机因素,采用SAS中的一般线性模型程序计算凤梨草莓（41份）、东北草莓（3份）、绿色草莓（4份）、黄毛草莓（3份）、种间杂交种（6份）的病情指数差异（P＜0.05）。【结果】 供试材料中没有对炭疽病完全免疫的材料,所有材料在接种果生炭疽菌4 d后均有不同程度的发病。对不同材料间的对比发现,感染炭疽病后草莓叶柄病斑长度与叶面病斑数量、叶片病斑直径均呈极显著正相关。以接种炭疽菌后的叶柄病斑长度为依据可以对供试材料的炭疽病抗性进行区分,供试材料中高抗17份、抗病20份、中抗21份、中感3份、感病6份、高感4份,炭疽病抗性水平为中抗及以上的材料占全部供试材料的81.7%。凤梨草莓中的‘3公主’‘森加森加拉’‘达赛莱克特’‘全明星’‘香野’‘威斯塔尔’和‘京藏香’,种间杂交种中的‘桃薰’对炭疽病的抗性水平为高抗。在草莓野生种中全部东北草莓、绿色草莓、东方草莓供试材料对炭疽病的抗性水平为高抗,东北草莓和绿色草莓的病情指数显著低于凤梨草莓。【结论】 对草莓属种质资源71份材料的炭疽病抗性进行了评价,从栽培草莓中筛选出‘3公主’‘森加森加拉’‘达赛莱克特’‘全明星’‘香野’‘威斯塔尔’‘京藏香’7个高抗品种。东北草莓和绿色草莓中存在高抗炭疽病的资源,其对炭疽病的整体抗性水平显著高于凤梨草莓,在种间杂交育种过程中可以作为炭疽病抗原来源。

关键词: 草莓, 种质资源, 果生炭疽菌, 炭疽病, 抗性

Abstract:

【Objective】 The objective of this study is to evaluate the anthracnose resistance level of 71 strawberry accessions including 24 accessions from 12 Fragaria species, 41 cultivars from Fragaria × ananassa and 6 interspecific hybrids, and to provide a basis for the utilization of strawberry germplasm resources in resistance breeding.【Method】 Colletotrichum fructicola isolate Gwha-1, which was isolated from strawberry petiole, was used as inoculation pathogen. The suspension of C. fructicola (1×10 ⁶ conidia/mL) was evenly inoculated on the leaf surface and petiole of strawberry leaves by in vitro inoculation method. Inoculated leaves were moisturized at 28℃ for 4 days, and then the incidence of each leaf was investigated. The lesion number on leaf surface, the largest lesion diameter on leaf surface, and the maximum lesion length on petiole were counted. Correlation analysis was performed on the lesion length on petiole, the lesion number and the lesion diameter on leaf surface of different materials using IBM SPSS 15.0 software. The disease severity of each leaf was ranked according to the lesion length on petiole. The disease index of each material was calculated, and the anthracnose resistance of the tested materials was evaluated based on the disease index of petiole after inoculation with C. fructicola isolate Gwha-1. The general linear model procedure (PROC GLM) in SAS was used to analyze the differential significance (P＜0.05) among five strawberry species, including F. × ananassa (41 accessions), F. mandschurica (3 accessions), F. viridis (4 accessions), F. nilgerrensis (3 accessions), and interspecific hybrids (6 accessions). Species were used as fixed factors and tested materials as random factors.【Result】 There was no completely immune material to anthracnose caused by C. fructicola in the tested materials, and all materials had different degrees of disease incidence after 4 days of inoculation with C. fructicola isolate Gwha-1. The comparison of different materials showed that the lesion length on leaf petiole was positively correlated with the lesion number on leaf and the lesion diameter on leaf surface after inoculation. According to the lesion length on petiole after inoculation with C. fructicola, the anthracnose resistance of the tested materials could be distinguished. The numbers of materials with anthracnose resistance level of high resistance, resistance, medium resistance, medium susceptibility, susceptibility and high susceptibility were 17, 20, 21, 3, 6 and 4, respectively, 81.7% of the tested materials had medium resistance, resistance or higher resistance level to anthracnose caused by C. fructicola. ‘3 Gongzhu’ ‘Senga Sengana’ ‘Darselect’ ‘Allstars’ ‘Kaorino’ ‘Veestar’ and ‘Jingzangxiang’ in the F. × ananassa, and ‘Tokun’ in interspecific hybrids had high resistance to anthracnose. In the wild species of strawberry, the resistance level of all the F. mandschurica, F. viridis and F. orientalis tested materials was high resistance, and the disease index of F. mandschurica and F. viridis was significantly lower than that of F. × ananassa. 【Conclusion】 The anthracnose resistance of 71 strawberry resources was evaluated. Seven cultivars of high resistance to anthracnose were selected from the cultivated strawberries, including ‘3 Gongzhu’ ‘Senga Sengana’ ‘Darselect’ ‘Allstar’ ‘Kaorino’ ‘Veestar’ and ‘Jingzangxiang’. There are high anthracnose resistant resources in F. mandschurica and F. viridis. Their resistance level to anthracnose is significantly higher than that of F. × ananassa, which can be used as an antigen source of anthracnose in interspecific hybridization.

Key words: strawberry, germplasm resources, Colletotrichum fructicola, anthracnose, resistance

韩永超,曾祥国,向发云,过聪,张庆华,陈丰滢,关伟. 草莓属植物种质资源对炭疽病抗性的离体评价[J]. 中国农业科学, 2019, 52(20): 3585-3594.

YongChao HAN,XiangGuo ZENG,FaYun XIANG,Cong GUO,QingHua ZHANG,FengYing CHEN,Wei GUAN. In vitro Evaluation of Strawberry Germplasm Resources for Resistance to Anthracnose[J]. Scientia Agricultura Sinica, 2019, 52(20): 3585-3594.

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图/表 4

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表1

草莓种质资源对炭疽病的抗性水平鉴定"

种类 Species	品种或编号 Cultivar or material	病情指数 Disease index	抗性水平 Resistance level	来源 Source
东北草莓F. mandschurica	HBYCM023	1.85	高抗HR	中国黑龙江Heilongjiang, China
	HBYCM009	2.47	高抗HR	中国吉林Jilin, China
	HBYCM016	4.94	高抗HR	中国黑龙江Heilongjiang, China
黄毛草莓F. nilgerrensis	HBYCM019	19.75	抗病R	中国四川Sichuan, China
	HBYCM015	28.40	中抗MR	中国贵州Guizhou, China
	HBYCM018	14.81	抗病R	中国云南Yunnan, China
绿色草莓F. viridis	HBYCM005	2.47	高抗HR	中国新疆Xinjiang, China
	HBYCM004	4.94	高抗HR	中国新疆Xinjiang, China
	HBYCM022	9.88	高抗HR	中国新疆Xinjiang, China
	HBYCM010	8.64	高抗HR	中国新疆Xinjiang, China
森林草莓F. vesca	HBYCM024	11.11	抗病R	日本Japan
森林草莓F. vesca	HBYCM013	13.58	抗病R	中国吉林Jilin, China
西藏草莓F. nubicola	HBYCM026	30.86	中抗MR	德国German
五叶草莓F. pentaphylla	HBYCM006	28.40	中抗MR	中国甘肃Gansu, China
五叶草莓F. pentaphylla	HBYCM003	34.57	中抗MR	中国四川Sichuan, China
东方草莓F. orientalis	HBYCM005	9.88	高抗HR	中国黑龙江Heilongjiang, China
	HBYCM007	2.47	高抗HR	中国黑龙江Heilongjiang, China
伞房草莓F. corymbosa	HBYCM017	100.00	高感HS	中国山西Shanxi, China
西南草莓 F. moupinensis	HBYCM021	11.11	抗病R	中国西藏Tibet, China
西南草莓 F. moupinensis	HBYCM002	11.11	抗病R	中国西藏Tibet, China
麝香草莓F. moschata	HBYCM014	11.11	抗病R	加拿大Canada
弗州草莓F. virginiana	HBYCM008	12.35	抗病R	美国America
弗州草莓F. virginiana	HBYCM020	11.11	抗病R	德国German
智利草莓F. chilioensis	HBYCM011	11.11	抗病R	美国America
种间杂交种 Interspecific hybrids	桃薰Tokun	6.17	高抗HR	日本Japan
	粉红佳人Fenhongjiaren	12.35	抗病R	中国China
	白草莓1号Baicaomei No.1	16.05	抗病R	中国China
	粉佳人Pink Beauty	23.46	中抗MR	中国China
	粉公主Pink Princess	28.40	中抗MR	中国China
	俏佳人Pretty Beauty	28.40	中抗MR	中国China
凤梨草莓F. × ananassa	3公主3 Gongzhu	4.94	高抗HR	中国China
	森加森加拉Senga Sengana	6.17	高抗HR	德国German
	达赛莱克特Darselect	8.64	高抗HR	法国France
	全明星Allstar	8.64	高抗HR	美国America
	威斯塔尔Veestar	8.64	高抗HR	加拿大Canada
	香野Kaorino	8.64	高抗HR	日本Japan
种类 Species	品种或编号 Cultivar or material	病情指数 Disease index	抗性水平 Resistance level	来源 Source
	京藏香Jingzangxiang	9.88	高抗HR	中国China
	甘露Ganlu	11.11	抗病R	中国China
	鬼努甘Kinuama	11.11	抗病R	日本Japan
	白雪公主Snow White	12.35	抗病R	中国China
	九天红韵Jiutianhongyun	12.35	抗病R	中国China
	红玉Hongyu	13.58	抗病R	中国China
	章姬Akihime	13.58	抗病R	日本Japan
	京桃香Jingtaoxiang	14.81	抗病R	中国China
	甜查理Sweet Charlie	18.52	抗病R	美国America
	07-D-5	20.99	中抗MR	中国China
	秀丽Xiuli	22.22	中抗MR	中国China
	丰香Toyonoka	23.46	中抗MR	日本Japan
	京留香Jingliuxiang	23.46	中抗MR	中国China
	京怡香Jingyixiang	23.46	中抗MR	中国China
	京郊小白Jingjiaoxiaobai	24.69	中抗MR	中国China
	晶悦Jingyue	25.93	中抗MR	中国China
	里瓦Riva	25.93	中抗MR	荷兰Netherlands
	艳丽Yanli	25.93	中抗MR	日本Japan
	京泉香Jinquanxiang	27.16	中抗MR	中国China
	特里拉Tenira	28.40	中抗MR	美国America
	晶玉Jingyu	30.86	中抗MR	中国China
	天香Tianxiang	30.86	中抗MR	中国China
	晶硕Jingshuo	38.93	中抗MR	中国China
	京凝香Jingningxiang	53.09	中感MS	中国China
	黔莓1号Qianmei No.1	55.56	中感MS	中国China
	幸香Sachinoka	55.56	中感MS	日本Japan
	雪里香Xuelixiang	60.49	感病S	韩国Korea
	越心Yuexin	65.43	感病S	中国China
	红颜Benihoppe	67.90	感病S	日本Japan
	宁丰Ningfeng	67.90	感病S	中国China
	硕丽Shuoli	67.90	感病S	中国China
	宁玉Ningyu	75.31	感病S	中国China
	晶瑶Jingyao	85.19	高感HS	中国China
	甜宝Tianbao	85.19	高感HS	中国China
	小白Xiaobai	85.19	高感HS	中国China

表1

表2

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种类 Species	倍性 Chromosome ploidy	份数 Number of accessions	叶面病斑数量 Lesion number on leaf surface	叶面病斑直径 Lesion diameter on leaf surface (cm)	叶柄病斑长度 Lesion length on petiole (cm)	叶柄病情指数 Disease index of petiole
东北草莓F. mandschurica	2x	3	0	0	0.03 (0.02-0.04)	3.09a (1.85-4.94)
黄毛草莓F. nilgerrensis	2x	3	0.93 (0-2.00)	0.24 (0-0.69)	0.27 (0.02-0.40)	20.99ab (14.81-28.40)
绿色草莓F. viridis	2x	4	0.06 (0-0.25)	0.02 (0-0.07)	0.05 (0.02-0.09)	6.48a (2.47-9.88)
森林草莓F. vesca	2x	2	0.51 (0.44-0.58)	0.29 (0.10-0.49)	0.11 (0.11-0.12)	12.35 (11.11-13.58)
西藏草莓F. nubicola	2x	1	2.00	0.28	0.24	30.86
五叶草莓F. pentaphylla	2x	2	0.94 (0.56-1.33)	0.09 (0.09-0.09)	0.38 (0.33-0.43)	31.48 (28.40-34.57)
东方草莓F. orientalis	4x	2	0.08 (0-0.17)	0.01 (0-0.01)	0.04 (0.02-0.07)	6.17 (2.47-9.88)
伞房草莓F. corymbosa	4x	1	3.50	2.22	5.62	100.00
西南草莓 F. moupinensis	4x	2	1.38 (0.44-2.31)	0.06 (0-0.11)	0.10 (0.09-0.10)	11.11 (11.11-11.11)
麝香草莓F. moschata	6x	1	0.11	0.01	0.11	11.11
弗州草莓F. virginiana	8x	2	0.11 (0-0.22)	0.01 (0-0.02)	0.13 (0.11-0.16)	11.73 (11.11-12.35)
智利草莓F. chilioensis	8x	1	1.08	0.47	0.14	11.11
凤梨草莓F. × ananassa	8x	41	1.34 (0-7.44)	0.10 (0-0.59)	0.46 (0.04-2.22)	33.75b (4.94-85.19)
种间杂交种Interspecific hybrids	8x, 10x	6	0.26 (0-0.89)	0.02 (0-0.05)	0.14 (0.07-0.20)	19.14ab (6.17-28.40)