水稻纹枯病生防木霉菌分离鉴定及适应性研究

doi:10.13304/j.nykjdb.2021.1079

摘要/Abstract

摘要：

为筛选对水稻纹枯病具有生防潜力的木霉菌，采用稀释涂布平板法从不同植物根际土样中分离纯化出20株木霉菌菌株。通过室内平板对峙培养试验，发现D1菌株及其发酵浓缩液对水稻纹枯病菌的抑制作用较强，抑菌率达分别达到了80.82%和69.55%。D1菌株对水稻安全，比对照增产27.41%；D1菌株对水稻纹枯病的防效范围为51.09%~70.64%。先用D1菌株处理24 h后接种纹枯病菌的防效最高，其次是纹枯病菌和D1菌株同时处理，先接病菌24 h后再用D1菌株处理的防效最低。说明D1菌株对纹枯病有预防和治疗作用，预防效果优于治疗。经形态学和分子生物学初步鉴定D1菌株为短密木霉（Trichoderma brevicompactum）。D1菌株与辣椒炭疽病菌等8种作物病原菌真菌对峙培养，对它们的生长抑菌率在63.03%~95.18%。在20~30 ℃和pH 4~5时D1菌株对纹枯病菌的抑菌效果最好。5种杀菌剂对D1菌株的菌丝生长毒力不同，EC₅₀值为0.042~384.24 mg·L^-1，在田间常用水平下均有较强的抑菌作用，抑菌率为75.8%~100%。研究结果为水稻纹枯病的无公害防治提供了理论依据。

关键词: 水稻纹枯病, 木霉菌, 抑菌作用, 生物防治, 适应性

Abstract:

In order to screen out the Trichoderma spp.with strong biocontrol effect on rice sheath blight， 20 strains were isolated and purified from different plant rhizosphere soil samples by the dilution coating plate method. It was found that D1 strain and the fermentation concentrated had strong inhibitory effects on rice sheath blight， through the indoor plate confrontation culture test， and the inhibitory rates reached 80.82% and 69.55%， respectively. From the pot disease prevention test of the D1 strain against rice sheath blight， it was concluded that the D1 strain was against rice plants. D1 strain was safe and increased the yield of rice by 27.41% compared with sterile water treatment； the control effect of D1 strain on rice sheath blight was between 51.09% and 70.64%. The control effect of germs was the highest when strain D1 was first treated for 24 h and then followed by simultaneous inoculation treatment， and the treatment had the lowest control effect with D1 strain after inoculation with the germs for 24 h. It showed that the D1 strain had preventive and therapeutic effects on rice sheath blight， and the preventive effect was better than the curative effect. The D1 strain was initially identified as Trichoderma brevicompactum through morphology and molecular studies. A confrontation experiment between D1 and 8 plant pathogens showed that the inhibitory rate of D1 strain against them was between 63.03% and 95.18%； at 20~30 ℃ and pH 4~5， strain D1 was effective against rice sheath blight， with the best antibacterial effect. the tested 5 fungicides had different virulence to the mycelial growth of the D1 strain， and their EC₅₀ value was 0.042~384.24 mg·L^-1， but they had strong antibacterial effect at the common concentration in the field with the inhibitory rate from75.8% to 100%. This study provided theoretical basis for the pollution-free control of rice sheath blight.

Key words: rice sheath blight, Trichoderma spp., bacteriostasis, biological control, adaptability

中图分类号:

S476+.1

金辉, 王伟, 颜尘栋, 王薇, 李熙英. 水稻纹枯病生防木霉菌分离鉴定及适应性研究[J]. 中国农业科技导报, 2022, 24(9): 139-148.

Hui JIN, Wei WANG, Chendong YAN, Wei WANG, Xiying LI. Isolation， Identification and Adaptability of Trichoderma spp. for Biocontrol of Rice Sheath Blight[J]. Journal of Agricultural Science and Technology, 2022, 24(9): 139-148.

图/表 11

参考文献 26

1	陈利峰,徐敬友.农业植物病理学(南方版)[M].北京:中国农业出版社, 2001: 105-107.
2	SING A, ROHILLA R, SINGH U, et al.. An improved inoeulation technique for sheath blight of rice caused by Rhizotonia solani [J]. Can. J. Plant Pathol., 2002, 24(1): 65-68.
3	檀根甲,王子迎.水稻纹枯病时间与空间生态位的研究[J].中国水稻科学,2002(02):85-87.
	TAN G J, WANG Z Y. Study on temporal and spatial niches of rice blight [J]. Chin. J. Rice Sci., 2002(02): 65-68.
4	夏汉炎.水稻纹枯病防治进展[J].南方农业,2021,15(27):12-14.
	XIA H Y. Progress in the prevention and control of rice blight [J]. South China Agric.,2021,15(27):12-14.
5	束震.哈茨木霉菌与复合芽孢杆菌对西瓜枯萎病的防治效果及其促进西瓜生长的作用[J].安徽农学报,2021,27(23):116-118.
	SHU Z. The effect of Trichoderma Harz and Bacillus complex on the prevention and control of watermelon wilt and its role in promoting the growth of watermelon [J]. Anhui Agric. Sci. Bull., 2021,27(23):116-118.
6	杨兴堂. 3株木霉菌的鉴定、生物学特性和抑菌能力及其对凤仙花的促生作用[D].哈尔滨:东北林业大学, 2016.
	YANG X T. Identification,biological characteristics and bacteriostatic ability of three Trichoderma strains and their pro-growth effect on hydrangea [D]. Harbin: Northeast Forestry University, 2016.
7	宋晓妍,孙彩云,陈秀兰,等.木霉生防作用机制的研究进展[J].中国农业科技导报,2006, 8 (6): 20-25.
	SONG X Y, SUN C Y, CHEN X L, et al.. Research advanceson mechanism of Trichoderma in biological control [J]. J. Agric. Sci. Technol., 2006, 8(6): 20-25.
8	田娜,姚瑞丽,姚艳平,等.生防木霉菌的分离和筛选[J].山西农业大学学报(自然科学版),2013,22(4):313-318.
	TIAN N, YAO R L, YAO Y P, et al..Isolation and screening of biocontrol Trichoderma [J]. J. Shanxi Agric. Univ. (Nat. Sci.),2013,22(4):313-318.
9	王艳丽,沈瑛,徐同.哈茨木霉防治水稻纹枯病研究[J].植物保护学报,2000(2):97-101.
	WANG Y L, SHEN Y, XU T. Study on Trichoderma harzianum strains to control of rice sheat blight [J]. J. Plant Protec.,2000(2):97-101.
10	董晓军,贾瑞芬.木霉菌防治番茄灰霉病的初步研究[J].基层农技推广,2018,6(01):33-35.
11	杨合同.木霉分类与鉴定[M].北京:中国大地出版社,2009:158-165.
12	董红刚,袁林泽,康晓霞,等.7种杀菌剂防治水稻纹枯病田间药效比较试验[J]. 上海农业科技,2015(6):124-125, 105.
	DONG H G, YUAN L Z, KANG X X, et al.. Comparative experiment on the efficacy of 7 fungicides in the field for the prevention and control of rice blight [J]. J. Shanghai Agric. Sci. Technol.,2015(6):124-125, 105.
13	孙广宁,宗兆峰.植物病理学实验技术[M].北京:中国农业出版社,2002:256-265.
14	BISSETT J. A revision of the genus Trichoderma Ⅰ‍. section longibrachiatum sect.nov. [J]. Can. J. Bot.,1984,62:924-931.
15	BISSETT J. Ar revision of the genus Trichoderma Ⅱ‍. infrageneric classification [J]. Can. J. Bot., 1991,69:2357-2372.
16	BISSETT J. A revision of the genus Trichoderma Ⅲ‍. section pachybasium [J]. Can. J. Bot.,1991,69:2373-2417.
17	BISSETT J. A revision of the genus Trichoderma Ⅳ.additional notes on section longibrachiatum [J]. Can. J. Bot.,1991,69:2418-2420.
18	郭润芳,刘晓光,高克祥,等.拮抗木霉菌在生物防治中的应用与研究进展[J].中国生物防治,2002,18(4):180-184.
	GUO R F, LIU X G, GAO K X, et al.. Application and research progress of antagonistic Trichoderma in biological control [J]. Chin. J. Biol. Control,2002,18(4):180-184.
19	陈泉,赵晓燕,李纪顺,等.木霉与土壤生态环境关系研究综述[J].台湾农业探索,2013(6):66-68.
	CHEN Q, ZHAO X Y, LI J S,et al..A review on the relationship between Trichoderma and soil ecological environment [J]. Taiwan Agric. Res., 2013(6):66-68.
20	申君,杨绍丽,谷清义,等.一株辣椒根腐病拮抗木霉菌Tb1的筛选与鉴定[J/OL].东北农业科学,2022:1-9 [2022-08-09]. .
	SHEN J, YANG S L, GU Q Y.et al.. Screening and identification of a pepper root rot antagonizing Trichoderma Tb1 [J/OL]. J. Northeast Agric. Sci., 2002:1-9[2022-08-09]. .
21	郭成,张小杰,张有富,等.短密木霉菌株GAS1-1的分离鉴定、拮抗作用及其生物学特性[J].植物保护学报,2019,46(2):305-312.
	GUO C, ZHANG X J, ZHANG Y Fet al.. Isolation, identification, antagonism and biological characteristics of Trichoderma brevis strain GAS1-1 [J]. J. Plant Protec.,2019,46(2):305-312.
22	吴紫燕,沈少力,糜芳,等.作物根围土壤木霉菌物种多样性及其体外拮抗病原菌效应[J].菌物研究,2017,15(3):177-182, 187.
	WU Z Y, SHEN S L, MI Fet al.. Species diversity of Trichoderma in the soil around crops and their antagonistic pathogenic effects in vitro [J]. J. Fungal Res.,2017,15(3):177-182, 187.
23	李松鹏.两株水稻根际木霉菌株生物学特性及生防潜能研究[D].武汉:华中农业大学, 2017.
	LI S P.Study on the biological characteristics and biocontrol potential of two rice rhizosphere Trichoderma strains [D]. Wuhan: Huazhong Agricultural Universit, 2017.
24	潘玮.绿色木霉厚垣孢子与分生孢子生物学特性及生防效果的比较研究[D].北京:中国农业科学院, 2006.
	PAN W. A comparative study on the biological characteristics and biocontrol effects of Trichoderma green trichoderma and conidia [D]. Beijing:Chinese Academy of Agricultural Sciences, 2006.
25	李雪婷,郑树仁,聂倩文,等.菌株S17-377的鉴定及其对水稻纹枯病防治作用机制[C]//中国植物病理学会2019年学术年会论文集.北京:中国农业科学技术出版社,2019:534.
	LI X T,ZHENG S R,NIE Q W, et al.. Identification of strain S17-377 and its mechanism of controlling rice sheath blight [C]// Proceedings of 2019 Academic Annual Meeting of Chinese Society of Plant Pathology. Beijing: China Agricultural Science and Technology Press, 2019:534.
26	张林.防治土传病害的几种主要化学农药对木霉菌厚垣孢子的影响[D].北京:中国农业科学院, 2014.
	ZHANG L.Effects of several major chemical pesticides for the control of soil-borne diseases on the pachycete spores of Trichoderma [D]. Beijing:Chinese Academy of Agricultural Sciences,2014.

杀菌剂 Pesticides	用量 Dosage/ （mg·L^-1）
1.5%多抗霉素 1.5% polyantimycin	1	5	10	50	100	500
75%百菌清 75% chlorothalonil	1	5	10	50	100	500
50%多菌灵 50% carbendazim	—	0.005	0.010	0.050	0.100	0.500
50 %腐霉利 50% pythium	—	0.005	0.010	0.050	0.100	0.500
80%代森锰锌 80% mancozeb	—	50	100	500	1 000	2 000

杀菌剂 Pesticides	用量 Dosage/ （mg·L^-1）
1.5%多抗霉素 1.5% polyantimycin	1	5	10	50	100	500
75%百菌清 75% chlorothalonil	1	5	10	50	100	500
50%多菌灵 50% carbendazim	—	0.005	0.010	0.050	0.100	0.500
50 %腐霉利 50% pythium	—	0.005	0.010	0.050	0.100	0.500
80%代森锰锌 80% mancozeb	—	50	100	500	1 000	2 000

根际土样 Soil sample of rhizosphere	真菌菌株数 Number of fungus	木霉菌菌株数 Number of Trichoderma strains	编号Code
玉米Maize	45	8	Y1~Y8
西红柿Tomato	9	1	X1
芹菜Celery	40	6	Q1~Q6
辣椒Pepper	40	4	L1~L4
丁香Clove	9	1	D1
杨树Poplar	13	0	—
榆树Elm	10	0	—
油松Pinus	15	0	—
苹果、梨Apple and peer	8	0	—

根际土样 Soil sample of rhizosphere	真菌菌株数 Number of fungus	木霉菌菌株数 Number of Trichoderma strains	编号Code
玉米Maize	45	8	Y1~Y8
西红柿Tomato	9	1	X1
芹菜Celery	40	6	Q1~Q6
辣椒Pepper	40	4	L1~L4
丁香Clove	9	1	D1
杨树Poplar	13	0	—
榆树Elm	10	0	—
油松Pinus	15	0	—
苹果、梨Apple and peer	8	0	—

菌株编号 Strain codes	菌落直径Colony diameter/cm		抑菌率Inhibitory rate/%
菌株编号 Strain codes	菌株 Strain	发酵浓缩液 Fermentation concentrated	菌株 Strain	发酵浓缩液 Fermentation concentrated
D1	2.10	3.17	80.82±2.30 a	69.55±2.57 a
Y7	2.40	8.33	76.71±0.00 b	5.76±4.99 f
Y5	2.50	8.27	75.34±1.37 bc	6.58±3.11 f
L2	2.60	8.43	73.97±1.37 c	4.53±1.89 f
L1	2.70	8.13	72.60±2.37 cd	8.23±4.34 f
L3	2.70	4.87	72.60±0.00 cd	48.56±5.15 c
Q4	2.70	7.33	72.60±1.37 cd	18.11±2.57 de
Q3	2.87	7.77	70.32±1.58 de	12.76±3.11 f
Q2	2.90	7.40	69.86±0.00 def	17.28±2.47 de
Q6	2.93	7.83	69.41±0.79 efg	11.93±5.70 f
Y8	3.03	8.43	68.04±0.79 efgh	4.53±1.89 f
Q5	3.03	4.27	68.04±2.09 efgh	55.97±1.89 bc
Y4	3.07	4.03	67.58±2.09 efghi	58.85±3.56 b
Y2	3.10	3.77	67.12±1.37 fghi	62.14±6.09 b
Y3	3.10	7.47	67.12±0.00 fghi	16.46±3.97 de
Y1	3.13	4.87	66.67±2.09 ghi	48.56±8.03 c
Q1	3.17	7.83	66.21±0.79 hi	11.93±2.57 ef
L4	3.17	4.23	66.21±1.58 hi	56.38±4.67 b
Y6	3.20	7.10	65.75±1.37 hi	20.99±7.98 d
X1	3.27	7.87	64.84±2.09 i	11.52±1.89 ef
CK	8.00	8.80	—	—