黄淮海夏玉米区玉米籽粒带菌检测分析

doi:10.3864/j.issn.0578-1752.2018.18.007

Abstract

Abstract:

【Objective】 The objective of this study is to determine the amount of fungi carried in maize grains in Huang-Huai- Hai summer maize region, and to provide reference for safe production, storage, processing and quarantine of maize. 【Method】A total of 720 maize ears were collected at the stages of milk-ripening and full-ripening in 90 cities or counties in 4 provinces (Hebei, Henan, Shandong, Anhui) of the Huang-Huai-Hai summer maize region, 4 full maize ears with no symptom on the surface were sampled in each city or county. All samples were tested for the amount and species of fungi carried, including external and internal tests of maize grains. Washing detection method was used to calculate the spores load on the grain surface, the spore load on the grain surface and isolation rate of each genus were calculated by counting the total number of colonies and dilution multiple. The PDA plate method was used for internal detection. Ten grains tested externally were sterilized and cultured on PDA plate. The fungi carrying rate of grains and the isolation frequency of each genus of fungi were calculated by counting the fungi carrying rate of each grain. In addition, the morphological and molecular identification of the higher frequency fungi were carried out. 【Result】The samples carried a large number of fungi. The spores load of the tested samples ranged from 0 to 1 886 per grain, with an average of 439 spores per grain at milk-ripening stage, the fungi-carrying rate of grains ranged from 0 to 65.0%, with an average of 23.6%. The spores load ranged from 18 to 2 658 per grain, with an average of 942 spores per grain at full-ripening stage, the fungi-carrying rate of grains ranged from 10.0% to 100.0%, with an average of 59.6%. The amount of fungi carried in full-ripening stage was higher than that in milk-ripening stage, but in some areas, the amount of fungi carried was still large in milk-ripening stage. There were differences in the amount of fungi carried in maize grains in different areas. The amount of fungi carried in maize grains in Henan Province was the largest, and Anhui Province had the least amount of fungi, Hebei Province and Shandong Province were in the middle and the difference was not significant. Fungal communities carried both inside and outside of maize grains included Fusarium spp., Penicillium spp., Aspergillus spp., Alternaria spp., Trichoderma spp., Rhizopus spp., Hel-minthosporium spp., Mucor spp. The isolation rate of Fusarium spp. in the grain external and internal detection was 59.1% and 36.1% respectively at milk-ripening stage, indicating that a large number of Fusarium spp. occurred at milk-ripening stage of maize. The isolation rate of Penicillium spp. and Aspergillus spp. in external grains was 8.9% and 0.7% respectively, and the isolation frequency in internal grains was 6.0% and 1.9% respectively, indicating that Penicillium spp. and Aspergillus spp. had begun to infect maize ear at milk-ripening stage. The isolation rate of Fusarium spp. in the grain external and internal detection was 71.9% and 58.5% respectively at full-ripening stage, the isolation rate of Penicillium spp. and Aspergillus spp. in external grains was 17.0% and 0.9% respectively, and the isolation frequency in internal grains was 9.3% and 2.6% respectively, indicating that Fusarium spp., Penicillium spp. and Aspergillus spp. were the main fungi carried by maize grains in Huang-Huai-Hai summer maize region. Morphological and molecular identification results showed that F. verticillioides was the dominant strain in Fusarium spp., the isolation frequency of F. verticillioides, F. proliferatum, F. graminearum was 29.7%, 25.9%, 1.3%, respectively. P. funiculosum and P. oxalicum were the dominant strains in Penicillium spp., and the isolation frequency was 5.0% and 3.6%, respectively. A. flavus and A. niger were the dominant strains in Aspergillus spp., and the isolation frequency was 1.4% and 1.2%, respectively. 【Conclusion】 The maize grains with no surface symptom carried a large number of fungi in both milk-ripening and full-ripening stages, and the amount of fungi carried in the full-ripening stage is higher than that in the milk-ripening stage. Fusarium spp. is the most frequently isolated fungi in Huang-Huai-Hai summer maize region and F. verticilliflora is the dominant strain. It is the dominant pathogen carried by local maize grains.

Key words: maize grain, fungi detection, isolation frequency, Huang-Huai-Hai summer maize region

LiuYan JIN, Ning GUO, Jie SHI, HaiJian ZHANG, ShuSen LIU, JiaQi ZHANG. Detection and Analysis of Fungi Carried by Maize Grain in Huang-Huai-Hai Summer Maize Region[J].Scientia Agricultura Sinica, 2018, 51(18): 3508-3519.

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Figures/Tables 6

Table 1

Fig. 1

Table 2

Results of grain carrying fungi in different areas"

编号 Number	省 Province	市/县 City/County	籽粒带菌率Rate of grains with fungi (%)		孢子负荷量（个/粒）Spore load
编号 Number	省 Province	市/县 City/County	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage
1	河北Hebei	武强Wuqiang	10.0	52.5	75	33
2	河北Hebei	涿州Zhuozhou	2.5	55.0	1448	2160
3	河北Hebei	万全Wanquan	47.5	47.5	188	228
4	河北Hebei	昌黎Changli	2.5	40.0	34	128
5	河北Hebei	怀来Huailai	5.0	47.5	0	83
6	河北Hebei	滦平Luanping	47.5	87.5	1064	1123
7	河北Hebei	廊坊Langfang	15.0	85.0	90	1780
8	河北Hebei	平泉Pingquan	35.0	77.5	240	1895
9	河北Hebei	玉田Yutian	32.5	75.0	604	863
10	河北Hebei	故城Gucheng	32.5	87.5	202	538
11	河北Hebei	鹿泉Luquan	5.0	72.5	1020	1278
12	河北Hebei	沧州Cangzhou	12.5	60.0	324	425
13	河北Hebei	定兴Dingxing	17.5	65.0	54	70
14	河北Hebei	衡水Hengshui	7.5	75.0	636	938
15	河北Hebei	赵县Zhaoxian	12.5	80.0	330	823
16	河北Hebei	邢台Xingtai	2.5	40.0	174	265
17	河北Hebei	藁城Gaocheng	16.0	42.5	77	313
18	河北Hebei	蠡县Lixian	37.5	50.0	984	1303
19	河北Hebei	高阳Gaoyang	17.5	42.5	66	288
20	河北Hebei	大明Daming	22.5	52.5	708	1263
21	河北Hebei	肃宁Suning	0.0	47.5	260	1453
22	河北Hebei	成安Cheng’an	7.5	65.0	86	1845
23	河北Hebei	无极Wuji	52.5	57.5	14	1008
24	河北Hebei	鸡泽Jize	15.0	50.0	68	1520
25	河北Hebei	新乐Xinle	5.0	37.5	642	648
26	河北Hebei	河间Hejian	20.0	50.0	64	1260
27	河北Hebei	满城Mancheng	42.5	55.0	36	78
28	河北Hebei	望都Wangdu	30.0	70.0	12	793
29	河北Hebei	深州Shenzhou	5.0	67.5	64	1183
30	河南Henan	睢阳Suiyang	2.5	52.5	28	265
31	河南Henan	新郑Xinzheng	27.5	100.0	2	275
32	河南Henan	汝阳Ruyang	20.0	57.5	112	360
33	河南Henan	温县Wenxian	25.0	97.5	88	573
34	河南Henan	淇县Qixian	10.0	72.5	42	168
35	河南Henan	南阳Nanyang	5.0	95.0	4	643
36	河南Henan	鹤壁Hebi	47.5	87.5	370	453
37	河南Henan	驻马店Zhumadian	47.5	52.5	78	545
38	河南Henan	淮阳Huaiyang	37.5	100.0	368	18
续表2 Continued Table 2
编号 Number	省 Province	市/县 City/County	籽粒带菌率Rate of grains with fungi (%)		孢子负荷量（个/粒）Spore load
编号 Number	省 Province	市/县 City/County	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage
39	河南Henan	遂平Suiping	10.0	97.5	492	867
40	河南Henan	夏邑Xiayi	60.0	82.5	452	1208
41	河南Henan	焦作Jiaozuo	37.5	92.5	106	1503
42	河南Henan	漯河Luohe	35.0	70.0	970	1910
43	河南Henan	通许Tongxu	20.0	48.0	160	2252
44	河南Henan	平顶山Pingdingshan	32.5	52.5	1410	2290
45	河南Henan	内黄Neihuang	22.5	45.0	160	783
46	河南Henan	襄县Xiangxian	15.0	77.5	1056	2658
47	河南Henan	洛阳Luoyang	32.5	70.0	1320	2638
48	河南Henan	太康Taikang	60.0	62.5	266	600
49	河南Henan	邓州Dengzhou	65.0	85.0	1632	1850
50	河南Henan	原阳Yuanyang	55.0	60.0	1074	1498
51	河南Henan	滑县Huaxian	30.0	47.5	846	953
52	河南Henan	南乐Nanle	12.5	47.5	920	1233
53	河南Henan	商丘Shangqiu	2.5	42.5	776	995
54	河南Henan	辉县Huixian	53.3	45.0	812	1185
55	河南Henan	新乡Xinxiang	22.5	45.0	156	400
56	山东Shandong	德州Dezhou	16.0	70.0	264	1038
57	山东Shandong	嘉祥Jiaxiang	30.0	52.5	770	808
58	山东Shandong	莘县Shenxian	47.5	55.0	621	805
59	山东Shandong	莱州Laizhou	27.5	47.5	302	660
60	山东Shandong	章丘Zhangqiu	20.0	52.5	640	1363
61	山东Shandong	菏泽Heze	32.5	45.0	339	1240
62	山东Shandong	泰安Taian	12.5	85.0	430	418
63	山东Shandong	冠县Guanxian	5.0	62.5	230	812
64	山东Shandong	莱阳Laiyang	0.0	77.5	1550	1836
65	山东Shandong	青岛Qingdao	22.5	45.0	613	932
66	山东Shandong	临沂Linyi	15.0	65.0	333	350
67	山东Shandong	济阳Jiyang	55.0	85.0	418	444
68	山东Shandong	齐河Qihe	50.0	72.5	410	936
69	山东Shandong	曲阜Qufu	25.0	57.5	270	366
70	山东Shandong	诸城Zhucheng	15.0	92.5	443	408
71	山东Shandong	平度Pingdu	20.0	32.5	86	635
72	山东Shandong	宁津Ningjin	30.0	47.5	456	1043
73	山东Shandong	莒县Juxian	12.5	47.5	34	2228
74	山东Shandong	郓城Yuncheng	27.5	20.0	306	850
75	山东Shandong	滨州Binzhou	42.5	62.5	12	605
76	山东Shandong	平原Pingyuan	27.5	42.5	882	978
77	山东Shandong	聊城Liaocheng	20.0	45.0	204	520
续表2 Continued Table 2
编号 Number	省 Province	市/县 City/County	籽粒带菌率Rate of grains with fungi (%)		孢子负荷量（个/粒）Spore load
编号 Number	省 Province	市/县 City/County	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage	乳熟期 Milk-ripening stage	完熟期 Full-ripening stage
78	山东Shandong	济宁Jining	20.0	42.5	1886	1948
79	山东Shandong	茌平Chiping	7.5	35.0	340	405
80	山东Shandong	平邑Pingyi	37.5	65.0	528	1308
81	山东Shandong	临朐Linqu	8.0	52.0	404	788
82	山东Shandong	潍坊Weifang	12.0	26.0	34	453
83	安徽Anhui	阜阳Fuyang	12.0	34.0	146	553
84	安徽Anhui	泗县Sixian	48.0	40.0	696	1233
85	安徽Anhui	立辛Lixin	18.0	64.0	224	775
86	安徽Anhui	蒙城Mengcheng	2.0	70.0	1250	1320
87	安徽Anhui	濉溪Suixi	6.0	70.0	18	308
88	安徽Anhui	蚌埠Bengbu	4.0	10.0	628	1458
89	安徽Anhui	界首Jieshou	40.0	54.0	284	290
90	安徽Anhui	宿州Suzhou	6.0	22.0	206	960
均值Average			23.6	59.6	439	942

Table 2

Table 3

Table 4

Table 5

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省份 Province	乳熟期采样时间 Sampling time of milk-ripening stage	完熟期采样时间 Sampling time of full-ripening stage
河北Hebei	2017-09-07—2017-09-09	2017-09-23—2017-09-25
河南Henan	2017-09-01—2017-09-03	2017-09-18—2017-09-20
山东Shandong	2017-09-04—2017-09-07	2017-09-21—2017-09-23
安徽Anhui	2017-09-04	2017-09-20

省份 Province	乳熟期分离比例 Isolation rate at milk-ripening stage (%)				完熟期分离比例 Isolation rate at full-ripening stage (%)
省份 Province	孢子负荷量（个/粒） Spore load	镰孢菌 Fusarium spp.	青霉菌 Penicillium spp.	曲霉菌 Aspergillus spp.	孢子负荷量（个/粒） Spore load	镰孢菌 Fusarium spp.	青霉菌 Penicillium spp.	曲霉菌 Aspergillus spp.
河北Hebei	330	58.5	6.5	0.6	882	58.7	22.9	0.9
河南Henan	527	62.6	8.6	0.2	1082	75.8	13.6	0.3
山东Shandong	474	61.4	6.5	1.9	895	83.3	11.6	2.2
安徽Anhui	432	53.7	13.8	0	862	69.7	19.9	0.2
均值Average	441	59.1	8.9	0.7	930	71.9	17.0	0.9

省份 Province	轮枝镰孢 F. verticillioides	层出镰孢 F. proliferatum	禾谷镰孢 F. graminearum	绳状青霉 P. funiculosum	草酸青霉 P. oxalicum	黄曲霉 A. flavus	黑曲霉 A. niger	其他 Others
河北Hebei	23.2	23.7	0.3	3.3	4.1	0.6	2.5	42.3
河南Henan	35.9	22.1	3.0	3.2	5.8	1.5	1.1	27.4
山东Shandong	31.4	30.1	1.7	2.1	4.5	1.3	1.1	27.8
安徽Anhui	28.1	27.5	0.0	11.5	0.0	2.0	0.0	30.9
均值Average	29.7	25.9	1.3	5.0	3.6	1.4	1.2	32.1

省份 Province	乳熟期分离频率 Isolation frequency at milk-ripening stage				完熟期分离频率 Isolation frequency at full-ripening stage
省份 Province	籽粒带菌率 Rate of grain with fungi	镰孢菌 Fusarium spp.	青霉菌 Penicillium spp.	曲霉菌 Aspergillus spp.	籽粒带菌率 Rate of grain with fungi	镰孢菌 Fusarium spp.	青霉菌 Penicillium spp.	曲霉菌 Aspergillus spp.
河北Hebei	19.3	34.6	8.9	1.3	59.8	49.2	7.9	3.1
河南Henan	30.3	40.7	4.0	3.1	68.7	63.0	10.0	2.7
山东Shandong	23.6	43.7	6.8	1.9	54.9	65.2	7.6	2.4
安徽Anhui	17.0	25.2	4.0	1.3	45.5	56.6	11.8	2.0
均值Average	22.6	36.1	6.0	1.9	57.2	58.5	9.3	2.6

Detection and Analysis of Fungi Carried by Maize Grain in Huang-Huai-Hai Summer Maize Region

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