四川玉米小斑病菌交配型组成与育性分析

doi:10.3864/j.issn.0578-1752.2021.12.006

Abstract

Abstract:

【Objective】The objective of this study is to clarify the distribution of mating types and fertility of Cochliobolus heterostrophus (anamorph: Bipolaris maydis), and to elucidate the possibility of the sexual reproduction of C. heterostrophus isolates in natural fields.【Method】A total of 544 C. heterostrophus isolates were collected from different geographical regions in Sichuan and Yunnan provinces during 2013-2018. The mating type of these isolates was detected by a multiple PCR method, and two standard tested isolates with high combining ability were screened by cross of MAT1-1 and MAT1-2 isolates. Then the 544 C. heterostrophus isolates were tested for their mating type and fertility with two standard test isolates. The fertility was determined by the development of pseudothecia and asci.【Result】Among the 544 isolates, 286 ones were identified as MAT1-1 and 258 as MAT1-2, accounting for 52.57% and 47.43%, respectively. Chi-square test gave a good fit of 1﹕1 ratio (χ²=1.441, P=0.230), indicating that these two mating types distributed in balance in natural fields. In addition, the investigation also showed that two mating types were present in all sampling locations and they had a basically same population structure in different years. After crossing tested isolates with standard isolates MSRS-2-3 (MAT1-1) and DY-12-1-2 (MAT1-2), the percentage of fertile and sterile isolates was 88.79% and 11.21%, respectively, and an obvious fertility differentiation was observed, which were characterized by 12.32% of high fertile, 27.39% of medium fertile, and 49.08% of low fertile isolates, respectively. Fertility structures of isolates varied in different geographical locations and in different collection years. The frequencies of fertile isolates ranged from 77.88% to 94.87% in 2013-2018. The percentage of fertile isolates in Southern Yunnan was highest up to 100%, followed by those in Northern Sichuan, Central Sichuan, Southern Sichuan and Eastern Sichuan regions, accounting for 93.25%, 89.87%, 83.33% and 79.31%, respectively. The percentage of fertile isolates in Western Sichuan region was lowest (69.23%).【Conclusion】MAT1-1 and MAT1-2 achieved an equilibrium in quantity among C. heterostrophus field populations, and the cross combinations between MAT1-1 and MAT1-2 isolates were generally fertile. Although sexual generation was rarely reported in C. heterostrophus population in natural condition, the current results indicated that cryptic sexual cycle might contribute to the genetic diversity of C. heterostrophus.

Key words: southern corn leaf blight, Cochliobolus heterostrophus, sexual reproduction, mating type, fertility, multiple PCR

SUN XiaoFang,LIU Min,PAN TingMin,GONG GuoShu. Mating Type and Fertility of Cochliobolus heterostrophus Causing Southern Corn Leaf Blight in Sichuan Province[J].Scientia Agricultura Sinica, 2021, 54(12): 2547-2558.

Figures/Tables 9

Table 1

Fig. 1

Fig. 2

Table 2

Table 3

Table 4

Fertility of C. heterostrophus isolates collected from 17 cities (prefectures) of Sichuan and Xishuangbanna of Yunnan provinces"

采样区 Sampling region		测试菌株数 Number of tested isolates	不同假囊壳形成能力的菌株数（出现频率） Number of isolates with different pseudothecium-forming levels (Frequency, %)				可育菌株数（出现频率）Number of fertile isolates (Frequency, %)	不同育性水平的菌株数（出现频率） Number of isolates with different fertility levels (Frequency, %)
采样区 Sampling region		测试菌株数 Number of tested isolates	高 High	中 Medium	低 Low	无 Absent	可育菌株数（出现频率）Number of fertile isolates (Frequency, %)	高 High	中 Medium	低 Low	不育 Sterile
四川中部 Central Sichuan	成都Chengdu	161	33 (20.50)	59 (36.65)	65 (40.37)	4 (2.48)	145 (90.06)	21 (13.04)	56 (34.78)	68 (42.24)	16 (9.94)
四川中部 Central Sichuan	眉山Meishan	66	11 (16.67)	20 (30.30)	29 (43.94)	6 (9.09)	59 (89.39)	6 (9.09)	19 (28.79)	34 (51.52)	7 (10.61)
总计Total		227	44 (19.38)	79 (34.80)	94 (41.41)	10 (4.41)	204 (89.87)	27 (11.89)	75 (33.04)	102 (44.93)	23 (10.13)
四川东部 Eastern Sichuan	内江Neijiang	11	0 (0)	4 (36.36)	7 (63.63)	0 (0)	11 (100.00)	0 (0)	4 (36.36)	7 (63.63)	0 (0)
	遂宁Suining	11	0 (0)	3 (27.27)	8 (72.73)	0 (0)	8 (72.73)	0 (0)	3 (27.27)	5 (45.45)	3 (27.27)
	南充Nanchong	16	4 (25.00)	3 (18.75)	8 (50.00)	1 (6.25)	13 (81.25)	3 (18.75)	3 (18.75)	7 (43.75)	3 (18.75)
	广安Guang’an	12	2 (16.67)	2 (16.67)	6 (50.00)	2 (16.67)	9 (75.00)	1 (8.33)	3 (25.00)	5 (41.67)	3 (25.00)
	巴中Bazhong	3	0 (0)	1 (33.30)	1 (33.30)	1 (33.30)	2 (66.67)	0 (0)	1 (33.33)	1 (33.33)	1 (33.33)
	达州Dazhou	5	0 (0)	1 (20.00)	2 (40.00)	2 (40.00)	3 (60.00)	0 (0)	1 (20.00)	2 (40.00)	2 (40.00)
总计Total		58	6 (10.34)	14 (24.14)	32 (55.17)	6 (10.34)	46 (79.31)	4 (6.90)	15 (25.86)	27 (46.55)	12 (20.69)
四川西部Western Sichuan	雅安Yaan	11	4 (36.36)	4 (36.36)	2 (18.18)	1 (9.09)	9 (81.82)	2 (18.18)	2 (18.18)	5 (45.45)	2 (18.18)
四川西部Western Sichuan	甘孜州Garze	2	0 (0)	0 (0)	0 (0)	2 (100.00)	0 (0)	0 (0)	0 (0)	0 (0)	2 (100.00)
总计Total		13	4 (30.77)	4 (30.77)	2 (15.38)	3 (23.08)	9 (69.23)	2 (15.38)	2 (15.38)	5 (38.46)	4 (30.77)
四川南部Southern Sichuan	自贡Zigong	5	0 (0)	5 (100.00)	0 (0)	0 (0)	5 (100.00)	0 (0)	0 (0)	5 (100.00)	0 (0)
	乐山Leshan	40	8 (20.00)	9 (22.5)	23 (57.50)	0 (0.00)	35 (87.50)	4 (10.00)	5 (12.50)	26 (65.00)	5 (12.50)
	宜宾Yibin	7	2 (28.57)	1 (14.29)	2 (28.57)	2 (28.57)	5 (71.43)	0 (0)	0 (0)	5 (71.43)	2 (28.57)
	泸州Luzhou	14	0 (0)	4 (28.57)	7 (50.00)	3 (21.43)	10 (71.43)	0 (0)	4 (28.57)	6 (42.86)	4 (28.57)
总计Total		66	10 (15.15)	19 (28.79)	32 (48.48)	5 (7.58)	55 (83.33)	4 (6.06)	9 (13.64)	42 (63.64)	11 (16.67)
四川北部 Northern Sichuan	德阳Deyang	84	15 (17.86)	35 (41.67)	32 (38.10)	2 (2.38)	80 (95.24)	9 (10.71)	26 (30.95)	45 (53.57)	4 (4.76)
	绵阳Mianyang	56	11 (19.64)	17 (30.36)	23 (41.07)	5 (8.93)	51 (91.07)	9 (16.07)	12 (21.43)	30 (53.57)	5 (8.93)
	广元Guangyuan	23	5 (21.74)	5 (21.74)	11 (47.83)	2 (8.70)	21 (91.30)	2 (8.70)	5 (21.74)	14 (60.87)	2 (8.70)
总计Total		163	31 (19.02)	57 (34.97)	66 (40.49)	9 (5.52)	152 (93.25)	20 (12.27)	43 (26.38)	89 (54.60)	11 (6.75)
云南南部 Southern Yunnan	西双版纳州Xishuangbanna	17	10 (58.82)	5 (29.41)	2 (11.76)	0 (0)	17 (100.00)	10 (58.82)	5 (29.41)	2 (11.76)	0 (0)

Table 4

Table 5

Table 6

Table 7

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假囊壳数量 Number of pseudothecia (dish)	假囊壳的形成能力等级 Pseudothecium-forming level	每个假囊壳中子囊的数量 Number of asci per pseudothecium	育性等级 Fertility level
＞200	高High	0	不育Sterile
		1-100	低Low
		101-200	中等Medium
		＞200	高High
100-200	中等 Medium	0	不育Sterile
		1-200	低Low
		＞200	中等Medium
1-100	低Low	0	不育Sterile
1-100	低Low	≥1	低Low
0	不育Sterile	—	不育Sterile

测试菌株 Tested isolate		MAT1-2
测试菌株 Tested isolate		9-10-7-2	A10	B73-2-2	C33	DY-12-1-2	CZ-1-2	S33-1-1	YN-1-1	YN-3-3	ZJ-10-2
MAT1-1	9-5-1	++	+		+	++			++	+	++
	C24	+	++^※	++	+^#		+	+	+	+	++
	MSRS-2-3	++	++		++
	CZ-10-2	++	++	++	+				++	++	++
	D4-4-1	++	++	++	+		++	++			++
	M17-1-2	++	++	++	++		++				++
	T91-1-2	++	++	++	++		++			++	++
	YN-3-2	++	++	++	++		++	++	++	++
	YN-6-1	++	++	++	++		++	++	++	++	++
	ZJ-7-1	++	++	++	++			++	++	++	++

采样年份 Sampling year	采样点数 Number of collection sites	测试菌株数 Number of tested isolates	MAT1-1菌株数Number of isolates of MAT1-1	MAT1-2菌株数Number of isolates of MAT1-2	MAT1-1﹕MAT1-2	可育菌株数Number of fertile isolates	可育菌株率 Percentage of fertile isolates (%)	不同育性水平的菌株数（出现频率） Number of isolates with different fertility levels (Frequency, %)
采样年份 Sampling year	采样点数 Number of collection sites	测试菌株数 Number of tested isolates	MAT1-1菌株数Number of isolates of MAT1-1	MAT1-2菌株数Number of isolates of MAT1-2	MAT1-1﹕MAT1-2	可育菌株数Number of fertile isolates	可育菌株率 Percentage of fertile isolates (%)	高 High	中 Medium	低 Low	不育 Sterile
2013	29	113	63	50	1.13﹕1	88	77.88	8 (7.08)	23 (20.35)	57 (50.44)	25 (22.12)
2014	9	28	14	14	1﹕1	22	78.57	4 (14.29)	8 (28.57)	10 (35.71)	6 (21.43)
2015	16	60	28	32	1﹕1.14	56	93.33	4 (6.67)	26 (43.33)	26 (43.33)	4 (6.67)
2016	18	39	17	22	1﹕1.29	37	94.87	4 (10.26)	11 (41.03)	22 (43.59)	2 (5.13)
2017	23	215	123	92	1.34﹕1	201	93.49	26 (12.09)	59 (27.44)	116 (53.95)	14 (6.51)
2018	8	89	41	48	1﹕1.17:	79	88.76	21 (23.60)	22 (24.72)	36 (40.45)	10 (11.24)
总计Total	103	544	286	258	1.11﹕1	483	88.79	67 (12.32)	149 (27.39)	267 (49.08)	61 (11.21)

测试菌株 Tested isolate		MAT1-1
测试菌株 Tested isolate		CZ-4-1	CZ-1-1	18LS-3-3	D4-1-3	18CZ-3-2	JY-2-1-1	YN-3-2	N1-1	18LS-3-10	GY-4-2-1
MAT1-2	B73-2-2		^※
	J-A	++	++	+^※	+	++	+			++	+
	YN-5-3
	DJY-1-2A	++	++	++	+		++				++
	MY-13-2-2			++	++
	18CZ-2-2
	ZJ-10-2	+		++	+	+	++	++	++		+
	MY-11-2-3				++
	CZ-5-2				++	++					++
	RS-1A				++		++		++

测试菌株 Tested isolate		MAT1-1
测试菌株 Tested isolate		XD-1-2-1	J1-3	WJ-1-3	GY-6-1-2	GY-1-1-1	D4-3-1	X3-1-1	MS-6-1-1	CZ-7-1-1	QL-1-2A	MS-7-1-1
MAT1-2	RS-2-2	+	+	++	+^#	+	+^#	++	+^#	++^#	+	++
	18LS-2-5	++	++	++	+^#	++	–	++	+		++	++
	YLS-3-3	+	+	++	+^#	+	–	+^#	+^#	+	+
	18LS-2-3	++	++	+	+^#	++^#	+^#	+	+^#	++^#	++^#	++^#
	GH-5-1-1	++^※	+	+	+^#	+^#	–	+^#	+^#	++^#	+	+
	P4-1-1	+	+	+	+^#	+^#	–	–	+^#	+^#	++^#	++
	ZJ-8-1	++	++	++	+^#	++	++^#	++	++^#	++		++
	JY-1-2-1	+	++	++	+^#	++	–	+	+^#	++	++	++

Mating Type and Fertility of Cochliobolus heterostrophus Causing Southern Corn Leaf Blight in Sichuan Province

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采样区 Sampling region		采样点数 Number of collection sites	测试菌株数Number of tested isolates	MAT1-1		MAT1-2		MAT1-1﹕MAT1-2
采样区 Sampling region		采样点数 Number of collection sites	测试菌株数Number of tested isolates	测试菌株数 Number of tested isolates	比例 Percentage (%)	测试菌株数Number of tested isolates	比例 Percentage (%)	MAT1-1﹕MAT1-2
四川中部 Central Sichuan	成都Chengdu	15	161	85	52.80	76	47.20	1.12﹕1
四川中部 Central Sichuan	眉山Meishan	11	66	39	59.09	27	40.91	1.44﹕1
总计Total		26	227	124	54.63	103	45.37	1.2﹕1
四川东部 Eastern Sichuan	内江Neijiang	4	11	7	63.64	4	36.36	1.75﹕1
	遂宁Suining	4	11	3	27.27	8	72.73	1﹕2.67
	南充Nanchong	6	16	11	68.75	5	31.25	2.2﹕1
	广安Guang’an	5	12	5	41.67	7	58.33	1﹕1.4
	巴中Bazhong	2	3	2	66.70	1	33.30	2﹕1
	达州Dazhou	3	5	2	40.00	3	60.00	1﹕1.5
总计Total		24	58	30	51.72	28	48.28	1.07﹕1
四川西部 Western Sichuan	雅安Yaan	9	11	6	54.55	5	45.45	1.2﹕1
四川西部 Western Sichuan	甘孜州Garze	1	2	2	100.00	0	0	/
总计Total		10	13	8	61.54	5	38.46	1.6﹕1
四川南部 Southern Sichuan	自贡Zigong	2	5	3	60.00	2	40.00	1.5﹕1
	乐山Leshan	8	40	23	57.50	17	42.50	1.35﹕1
	宜宾Yibin	3	7	5	71.43	2	28.57	2.5﹕1
	泸州Luzhou	3	14	6	42.86	8	57.14	1﹕1.33
总计Total		16	66	37	56.06	29	43.94	1.28﹕1
四川北部 Northern Sichuan	德阳Deyang	8	84	36	42.86	48	57.14	1﹕1.33
	绵阳Mianyang	11	56	29	51.79	27	48.21	1.07﹕1
	广元Guangyuan	7	23	15	65.22	8	34.78	1.88﹕1
总计Total		26	163	80	49.08	83	50.92	1﹕1.04
云南南部 Southern Yunnan	西双版纳州Xishuangbanna	1	17	7	41.18	10	58.82	1﹕1.43

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