一株对番茄枯萎病有拮抗作用的枯草芽孢杆菌的ARTP诱变与筛选

doi:10.11924/j.issn.1000-6850.casb20190800545

摘要/Abstract

摘要：

旨在利用常压室温等离子体(ARTP)诱变技术获得番茄枯萎病高效拮抗突变株。以一株对番茄枯萎病有拮抗作用的枯草芽孢杆菌N为初始菌株进行诱变处理,通过检测抑菌圈进行突变株筛选,并通过盆栽实验初步研究防效。通过拮抗带筛选以及抑菌圈复筛选出5株正突变菌株,其中正突变幅度最高的菌株为62,较初始菌株的抑菌圈直径提高了31.43%。经过6次传代,证实该突变株具有良好的遗传稳定性。盆栽实验表明,突变株62对于番茄枯萎病的防效达到75.76%。通过ARTP诱变获得的突变株稳定性好、拮抗效果提高明显,在防治番茄枯萎病方面有很好的应用前景。

关键词: 诱变育种, 枯草芽孢杆菌, 番茄枯萎病, 常压室温等离子体(ARTP), 盆栽防效

Abstract:

Atmospheric and room temperature plasma (ARTP) technique was used to obtain mutants with enhanced antagonistic activity against tomato fusarium wilt. Bacillus subtilis N which had antagonistic effect on tomato fusarium wilt was chosen as the initial strain. Detection of inhibition zone was carried out to screen mutants and the control effect of the positive strains against tomato fusarium wilt was investigated via pot experiment. Five positive mutants were obtained through antagonistic zone screening and second screening of inhibition zone, in which the strain 62 showed the highest positive-mutation range, the diameter of inhibition zone was increased by 31.43% compared with the original strain. After 6 times of consecutive subculturing, the strain 62 showed good genetic stability. In the pot experiment, the prevention effect of the strain 62 on tomato fusarium wilt reached 75.76%. The selected strain 62 treated by ARTP is genetically stable, and shows significantly improved antagonistic activity, thus it is believed that this strain has a great application potential in the control of tomato fusarium wilt.

Key words: mutagenesis breeding, Bacillus subtilis, tomato fusarium wilt, atmospheric and room temperature plasma (ARTP), control effect of pot experiment

中图分类号:

S435.79

张心青, 张萧萧, 杨传伦, 郭南南, 马娜娜, 蔡倩倩, 王秀芝. 一株对番茄枯萎病有拮抗作用的枯草芽孢杆菌的ARTP诱变与筛选[J]. 中国农学通报, 2020, 36(26): 44-49.

Zhang Xinqing, Zhang Xiaoxiao, Yang Chuanlun, Guo Nannan, Ma Nana, Cai Qianqian, Wang Xiuzhi. Bacillus subtilis Against Tomato Fusarium Wilt: ARTP Mutagenesis and Screening[J]. Chinese Agricultural Science Bulletin, 2020, 36(26): 44-49.

图/表 7

参考文献 20

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批次	分离菌株数	正突变菌株数	正突变率/%
1	100	34	34.00
2	100	37	37.00
3	150	59	39.33
4	150	60	40.00
5	144	54	37.50

批次	分离菌株数	正突变菌株数	正突变率/%
1	100	34	34.00
2	100	37	37.00
3	150	59	39.33
4	150	60	40.00
5	144	54	37.50

序号	正突变幅度/%	序号	正突变幅度/%	序号	正突变幅度/%	序号	正突变幅度/%
5	1.33±0.11	59	8.67±0.15	117	0.79±0.08	179	0.02±0.01
6	4.00±0.21	60	6.43±0.42	119	1.33±0.14	180	7.83±0.21
7	2.00±0.06	61	12.08±0.70	122	0.67±0.01	181	4.70±0.13
8	4.00±0.10	^?62	31.43±0.27	124	3.30±0.02	184	6.27±0.21
13	1.33±0.15	64	5.63±0.04	126	10.54±0.08	^?190	16.42±0.25
16	0.67±0.15	65	5.63±0.17	128	11.20±0.14	^?191	16.42±0.07
18	0.67±0.06	66	7.24±0.10	130	0.67±0.07	192	7.83±0.11
25	3.33±0.15	67	12.08±0.70	131	0.01±0.14	193	4.70±0.07
26	4.00±0.07	68	1.60±0.00	135	13.83±0.11	197	8.61±0.05
27	10.00±0.10	69	3.21±0.03	136	13.83±0.14	198	7.05±0.02
28	11.33±0.15	70	2.40±0.04	^?137	21.07±0.16	199	10.17±0.12
33	2.67±0.06	71	5.63±0.04	138	9.88±0.02	201	2.36±0.01
34	4.00±0.09	72	1.60±0.00	139	2.65±0.05	202	7.05±0.00
35	0.67±0.15	74	3.21±0.09	140	0.01±0.00	204	9.39±0.06
36	1.33±0.03	76	6.43±2.42	143	2.65±0.00	205	4.70±0.02
37	10.00±0.20	77	8.85±0.00	144	1.99±0.07	206	9.39±0.03
38	0.67±0.05	78	7.24±0.40	145	1.33±0.04	207	6.27±0.04
39	3.33±0.15	79	8.05±0.29	152	0.01±0.00	208	5.49±0.03
40	12.67±0.15	80	3.21±0.12	156	1.99±0.02	209	8.61±0.21
41	1.33±0.15	82	3.21±0.09	159	7.91±0.06	210	1.58±0.00
42	0.67±0.05	83	1.60±0.02	160	6.59±0.04	213	9.39±0.02
43	0.67±1.15	85	2.40±0.14	161	7.25±0.02	214	0.02±0.00
45	12.00±0.00	86	2.40±0.01	162	1.33±0.06	217	0.80±0.02
46	5.33±0.11	88	0.79±0.00	163	3.30±0.11	220	1.58±0.00
47	11.73±0.46	91	0.79±0.00	164	3.96±0.08	222	10.17±0.04
48	11.33±0.15	92	1.60±0.00	165	1.33±0.06	223	4.70±0.11
49	1.33±0.31	93	9.66±0.24	174	2.65±0.09	224	4.70±0.04
^?51	18.00±0.20	94	8.85±0.28	175	7.91±0.06	225	8.61±0.03
55	2.67±0.06	96	1.60±0.01	176	1.33±0.00	227	0.02±0.00
57	1.33±0.31	97	0.79±0.00	178	4.70±0.06	230	3.14±0.01

序号	正突变幅度/%	序号	正突变幅度/%	序号	正突变幅度/%	序号	正突变幅度/%
5	1.33±0.11	59	8.67±0.15	117	0.79±0.08	179	0.02±0.01
6	4.00±0.21	60	6.43±0.42	119	1.33±0.14	180	7.83±0.21
7	2.00±0.06	61	12.08±0.70	122	0.67±0.01	181	4.70±0.13
8	4.00±0.10	^?62	31.43±0.27	124	3.30±0.02	184	6.27±0.21
13	1.33±0.15	64	5.63±0.04	126	10.54±0.08	^?190	16.42±0.25
16	0.67±0.15	65	5.63±0.17	128	11.20±0.14	^?191	16.42±0.07
18	0.67±0.06	66	7.24±0.10	130	0.67±0.07	192	7.83±0.11
25	3.33±0.15	67	12.08±0.70	131	0.01±0.14	193	4.70±0.07
26	4.00±0.07	68	1.60±0.00	135	13.83±0.11	197	8.61±0.05
27	10.00±0.10	69	3.21±0.03	136	13.83±0.14	198	7.05±0.02
28	11.33±0.15	70	2.40±0.04	^?137	21.07±0.16	199	10.17±0.12
33	2.67±0.06	71	5.63±0.04	138	9.88±0.02	201	2.36±0.01
34	4.00±0.09	72	1.60±0.00	139	2.65±0.05	202	7.05±0.00
35	0.67±0.15	74	3.21±0.09	140	0.01±0.00	204	9.39±0.06
36	1.33±0.03	76	6.43±2.42	143	2.65±0.00	205	4.70±0.02
37	10.00±0.20	77	8.85±0.00	144	1.99±0.07	206	9.39±0.03
38	0.67±0.05	78	7.24±0.40	145	1.33±0.04	207	6.27±0.04
39	3.33±0.15	79	8.05±0.29	152	0.01±0.00	208	5.49±0.03
40	12.67±0.15	80	3.21±0.12	156	1.99±0.02	209	8.61±0.21
41	1.33±0.15	82	3.21±0.09	159	7.91±0.06	210	1.58±0.00
42	0.67±0.05	83	1.60±0.02	160	6.59±0.04	213	9.39±0.02
43	0.67±1.15	85	2.40±0.14	161	7.25±0.02	214	0.02±0.00
45	12.00±0.00	86	2.40±0.01	162	1.33±0.06	217	0.80±0.02
46	5.33±0.11	88	0.79±0.00	163	3.30±0.11	220	1.58±0.00
47	11.73±0.46	91	0.79±0.00	164	3.96±0.08	222	10.17±0.04
48	11.33±0.15	92	1.60±0.00	165	1.33±0.06	223	4.70±0.11
49	1.33±0.31	93	9.66±0.24	174	2.65±0.09	224	4.70±0.04
^?51	18.00±0.20	94	8.85±0.28	175	7.91±0.06	225	8.61±0.03
55	2.67±0.06	96	1.60±0.01	176	1.33±0.00	227	0.02±0.00
57	1.33±0.31	97	0.79±0.00	178	4.70±0.06	230	3.14±0.01

处理组	发病率/%	病情指数/%	防效/%
空白对照	100.00^a	62.50^a	/
药剂对照	80.00^b	18.97^b	69.65^a
菌剂组	60.00^c	15.15^c	75.76^b