水稻穗腐病菌强致病力且高产伏马菌素菌株筛选

doi:10.16819/j.1001-7216.2018.8045

摘要/Abstract

摘要：

目的旨在确立稳定高效的水稻穗腐病人工接种技术,鉴定不同菌株致病力。利用已有的伏马菌素检测方法分析测定两种培养基中不同菌株的伏马菌素合成能力,基于上述方法筛选强致病力高产伏马菌素菌株。方法分别在水稻孕穗期和抽穗期采用注射法和喷雾法接种,观察不同时期、不同接种方法下层出镰刀菌的致病性和稳定性;利用适合的接种方法和液相色谱-串联质谱检测法比较不同菌株的致病力和产毒能力。结果在花粉母细胞减数分裂期-花粉母细胞成熟期采用注射法接种水稻穗腐病,发病率较高且稳定;在花粉母细胞形成期-始穗期接种对水稻产量影响较大。基于该方法成功筛选出了强致病力菌株FP4、FP6、FP8、FP9、FP10;经HPLC-MS/MS分析测定了层出镰刀菌产生伏马菌素能力,获得了强致病力高产毒菌株FP4和FP9。稻谷培养基比玉米培养基更适合层出镰刀菌合成伏马菌素。结论水稻穗腐病初侵染为花粉母细胞形成期-花粉母细胞减数分裂期,最佳侵染时期为花粉母细胞形成期-齐穗期;穗腐病对水稻产量的影响与感病时期密切相关。

关键词: 水稻穗腐病, 层出镰刀菌, 人工接种, 伏马菌素, 致病力, 液相色谱-串联质谱法

Abstract:

【Objective】 The objective is to develop a stable and efficient artificial inoculation technique to identify the pathogenicity of different strains for rice spikelet rot disease (RSRD), and to test the ability of F. proliferatum to synthesize fumonisin in two culture media by detecting and analyzing the contents of fumonisin. Based on the methods, strains with high fumonisin production and strong pathogenicity were selected. 【Method】 Injection and spraying inoculation methods were adopted at booting and heading stages. The pathogenicity and stability of different inoculation methods at different growing stage of rice were investigated. The pathogenicity and the ability of fumonisin production of different strains were investigated by using suitable inoculation methods and HPLC-MS/MS analysis. 【Result】 The incidence of RSRD is high and stable with injection inoculation at pollen mother cell meiosis to maturing stages, and rice grain yield was significantly affected by inoculation during pollen mother cell formation to initial heading stage. The high pathogenic strains FP4, FP6, FP8, FP9 and FP10 were selected based on injection inoculation. The fumonisin synthesis assaying was conducted by using HPLC-MS/MS, and the strains FP4 and FP9, with high fumonisin production and strong pathogenicity were identified finally. The culture media made of rice grains was more suitable than that making of corn for fumonisin synthesis.【Conclusion】 The invasion period of F. proliferatum of RSRD was pollen mother cell formation to full heading stage, the initial infection stage was pollen mother cell formation to meiosis stage, and the effect of RSRD on rice yield was closely related to infection stage.

Key words: rice spikelet rot disease, Fusarium proliferatum, artificial inoculation, fumonisin, pathogenicity, HPLC-MS/MS

中图分类号:

S435.111.4⁺6

孙磊, 王玲, 刘连盟, 侯雨萱, 黎起秦, 黄世文. 水稻穗腐病菌强致病力且高产伏马菌素菌株筛选[J]. 中国水稻科学, 2018, 32(6): 610-616.

Lei SUN, Ling WANG, Lianmeng LIU, Yuxuan HOU, Qiqin LI, Shiwen HUANG. Screening for Strains of Rice Spikelet Rot Disease Pathogenic Fungus with High Fumonisin Production and Strong Pathogenicity[J]. Chinese Journal OF Rice Science, 2018, 32(6): 610-616.

图/表 8

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生育期 Rice growth stage	结实率 Seed setting rate/%	千粒重 1000-grain weight/g	穗重 Panicle weight/g
清水 Clean water	92.42±3.10 a	21.06±0.22 a	3.45±0.22 a
颖花分化期-雌雄蕊分化形成期 Spikelet differentiation to pistil and stamen formation stage(Ⅰ)	91.46±2.54 a	21.32±0.21 a	3.42±0.26 ab
花粉母细胞形成期-花粉母细胞减数分裂期 Pollen mother cell formation to meiosis(Ⅲ)	21.27±1.90 d	14.71±0.31 f	2.18±0.14 c
花粉母细胞减数分裂期-花粉母细胞成熟期 Pollen mother cell meiosis to maturing stage(Ⅳ)	24.14±2.03 d	15.14±0.18 e	2.23±0.09 c
破口期-始穗期 Rupturing to initial heading(Ⅴ)	24.15±3.06 d	16.41±0.26 d	2.41±0.35 c
始穗期-抽穗盛期 Initial heading to heading(Ⅵ)	53.79±2.04 c	17.46±0.16 c	2.96±0.31 b
抽穗盛期-齐穗期 Heading to full heading(Ⅶ)	83.56±3.16 b	18.32±0.21 b	3.27±0.42 ab

生育期 Rice growth stage	结实率 Seed setting rate/%	千粒重 1000-grain weight/g	穗重 Panicle weight/g
清水 Clean water	92.42±3.10 a	21.06±0.22 a	3.45±0.22 a
颖花分化期-雌雄蕊分化形成期 Spikelet differentiation to pistil and stamen formation stage(Ⅰ)	91.46±2.54 a	21.32±0.21 a	3.42±0.26 ab
花粉母细胞形成期-花粉母细胞减数分裂期 Pollen mother cell formation to meiosis(Ⅲ)	21.27±1.90 d	14.71±0.31 f	2.18±0.14 c
花粉母细胞减数分裂期-花粉母细胞成熟期 Pollen mother cell meiosis to maturing stage(Ⅳ)	24.14±2.03 d	15.14±0.18 e	2.23±0.09 c
破口期-始穗期 Rupturing to initial heading(Ⅴ)	24.15±3.06 d	16.41±0.26 d	2.41±0.35 c
始穗期-抽穗盛期 Initial heading to heading(Ⅵ)	53.79±2.04 c	17.46±0.16 c	2.96±0.31 b
抽穗盛期-齐穗期 Heading to full heading(Ⅶ)	83.56±3.16 b	18.32±0.21 b	3.27±0.42 ab

菌种 Strain	接种后14 d 14 d after inoculation			接种后21 d 21 d after inoculation
	病情指数 Disease index/%	病害严重度 Disease severity		病情指数 Disease index/%	病害严重度 Disease severity
	病情指数 Disease index/%	病害严重度 Disease severity	FP1	病情指数 Disease index/%	病害严重度 Disease severity	1.80±0.47 g	低 Low	2.98±0.27 ef	低 Low
FP2	3.72±0.87 de	中 Medium		5.16±0.73 cd	中 Medium
FP3	2.81±0.44 ef	低 Low		3.81±0.73 de	中 Medium
FP4	5.71±1.40 c	高 High		8.21±1.11 a	高 High
FP5	4.84±0.61 cd	中 Medium		4.58±0.31 cd	中 Medium
FP6	8.34±0.34 a	高 High		8.62±0.45 a	高 High
FP7	2.03±0.61 fg	低 Low		2.12±0.25 f	低 Low
FP8	4.86±0.44 cd	中 Medium		6.87±0.92 ab	高 High
FP9	8.47±0.48 a	高 High		9.29±0.53 ab	高 High
FP10	6.90±0.38 b	高 High		6.96±0.66 bc	中 Medium
FP11	4.51±0.34 d	中 Medium		4.63±1.23 cd	中 Medium
FP12	3.09±0.35 ef	低 Low		3.68±0.22 de	中 Medium

菌种 Strain	接种后14 d 14 d after inoculation			接种后21 d 21 d after inoculation
	病情指数 Disease index/%	病害严重度 Disease severity		病情指数 Disease index/%	病害严重度 Disease severity
	病情指数 Disease index/%	病害严重度 Disease severity	FP1	病情指数 Disease index/%	病害严重度 Disease severity	1.80±0.47 g	低 Low	2.98±0.27 ef	低 Low
FP2	3.72±0.87 de	中 Medium		5.16±0.73 cd	中 Medium
FP3	2.81±0.44 ef	低 Low		3.81±0.73 de	中 Medium
FP4	5.71±1.40 c	高 High		8.21±1.11 a	高 High
FP5	4.84±0.61 cd	中 Medium		4.58±0.31 cd	中 Medium
FP6	8.34±0.34 a	高 High		8.62±0.45 a	高 High
FP7	2.03±0.61 fg	低 Low		2.12±0.25 f	低 Low
FP8	4.86±0.44 cd	中 Medium		6.87±0.92 ab	高 High
FP9	8.47±0.48 a	高 High		9.29±0.53 ab	高 High
FP10	6.90±0.38 b	高 High		6.96±0.66 bc	中 Medium
FP11	4.51±0.34 d	中 Medium		4.63±1.23 cd	中 Medium
FP12	3.09±0.35 ef	低 Low		3.68±0.22 de	中 Medium

菌株 Strain	FB1含量 FB1 content/(mg·kg^-1)		FB2含量 FB2 content/(mg·kg^-1)
菌株 Strain	玉米 Corn kernel	稻谷 Paddy rice	玉米 Corn kernels	稻谷 Paddy rice
FP4	54.49±1.46 a	119.98±1.60 a	22.75±0.03 a	42.17±0.56 b
FP6	18.46±0.41 d	57.04±0.82 c	5.36±0.14 c	31.85±0.87 c
FP8	0.75±0.03 e	15.83±0.07 d	0.23±0.00 d	10.39±0.13 e
FP9	33.52±0.28 b	118.16±0.55 a	10.43±0.12 b	45.99±0.48 a
FP10	24.84±0.68 c	63.88±1.84 b	10.53±0.43 b	27.21±1.12 d