甘肃、青海地区小麦条锈菌监测及群体遗传多样性分析

doi:10.3864/j.issn.0578-1752.2020.18.006

Abstract

Abstract:

【Objective】In order to clarify the relationship about migration and inoculum source exchange of Puccinia striiformis f. sp. tritici (Pst) in spring epidemic time in Gansu and Qinghai provinces, and provide a theoretical basis for the epidemic prediction of wheat stripe rust and tracing the initial over-summering inoculum source and benefiting the effective disease management, the population genetic diversities and the potential spring epidemic migration of Pst were investigated. 【Method】The regions where stripe rust occurred frequently in the past years were selected as the investigation and studying regions. Four test sites were included in Gansu Province: Wenxian in Longnan City; Kongtong District in Pingliang City, Longdong; Linxia County, and Lintao County in Dingxi City, Longzhong; Two test sites were included in Qinghai Province: Chengbei District, Xining City; Huzhu County, Haidong City. In autumn of 2017, 82 wheat materials for trap nurseries were seeded sequentially in six regions according to the local wheat planting date in Gansu and Qinghai provinces. Field investigation was performed and molecular genotyping for 551 Pst samples collected from 82 materials with known resistant genes in six test sites of Gansu and Qinghai provinces from April to August 2018 was carried out by using 15 pairs of primers simple sequence repeat (SSR) markers. The GenAlEx and POPPR v2.5.0 software were used to analyze the data and the non-significant rbarD value might indicate linkage equilibrium was used to infer whether sexual recombination does occur or not. 【Result】The severity of the 82 wheat materials for trap nurseries of Pst in Gansu was more severe than that in Qinghai Province. The 15 SSR markers produced a total of 81 polymorphic alleles, ranging from 2 to 12 for each of the locus. A total of 505 unique multi locus genotypes (MLGs) were identified after clone correction cross the 551 Pst individuals, 32 MLGs were cloned and resampled between 2 and 6 times at six test sites. The entire genetic diversity (G) was 0.917 in six test sites from two provinces, the G value of Pingliang population (0.974) from Gansu was the highest, then it was Huzhu from Qinghai (0.957), and the Lintao population from Gansu was the lowest (0.841). The genetic variations of Pst mainly existed within the population. In spring epidemic, the migration occurred frequently among the populations. The gene flow was higher between Qinghai and Pingliang/Linxia than that was between Qinghai and Wenxian. The minimum spanning networks (MSN) and non-parametric discriminant analyses of principal components (DAPC) showed that the Huzhu and Xining populations from Qinghai had the closest relationship for the source of inoculum with the Pingliang and Linxia populations from Gansu, and the differences were the smallest. The genetic distance from the Lintao population was relatively far and the Lintao population was relatively independent. The Wenxian population was a completely independent population, with the largest difference from the other five populations. The analysis of linkage disequilibrium showed the presence of insignificant rbarD values in the Wenxian and Linxia populations from Gansu, and Xining population from Qinghai. The Wenxian population (rbarD=0.0139, P=0.186) showed obvious sexual recombination characteristics with a significant rbarD value. 【Conclusion】In spring epidemic season, the Pst source in Qinghai might originate from both Pingliang/Linxia and Wenxian, but it was mainly from Pingliang/Linxia. Signs for sexual reproduction were found in populations from Linxia and Wenxian of Gansu and Xining of Qinghai. The sexual reproduction might contribute to the richness of genetic diversity of Pst in Gansu and Qinghai.

Key words: Puccinia striiformis f. sp. tritici (Pst), wheat stripe rust, population genetic diversity, gene flow, sexual reproduction

HUANG MiaoMiao,CHEN WanQuan,CAO ShiQin,SUN ZhenYu,JIA QiuZhen,GAO Li,LIU Bo,LIU TaiGuo. Surveillance and Genetic Diversity Analysis of Puccinia striiformis f. sp. tritici in Gansu and Qinghai Provinces[J].Scientia Agricultura Sinica, 2020, 53(18): 3693-3706.

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Figures/Tables 9

Table 1

The collection information of the 551 Pst samples from six regions in Gansu and Qinghai provinces and the incidence results under the natural infection by Pst"

试验点 Test site		文县 Wenxian	平凉（崆峒区） Pingliang (Kongtong)	临夏 Linxia	临洮 Lintao	互助 Huzhu	西宁（城北区） Xining (Chengbei)
播种时间（年-月-日） Day of seeding time (Year-month-day)		2017-10-08	2017-09-25	2017-09-28	2017-10-08	2018-04-04	2018-04-25
采集时间（年-月-日） Day of collection (Year-month-day)		2018-04-14	2018-05-24	2018-06-16	2018-06-18	2018-08-08	2018-08-09
经度 Longitude		104°53′17″	106°40′11″	103°11′13″	103°31′18″	101°53′24″	101°44′56″
纬度 Latitude		32°50′43″	35°33′06″	35°36′49″	35°13′24″	36°42′41″	36°43′15″
海拔 Elevation (m)		786	1190	2010	1893	2360	2350
样本数量 Number of samples		27	132	156	178	47	11
编号 Code	品种名称 Cultivar	IT/S/I (N)
1	水源11 Suwon 11	3/5/T (1)	3/5/T (7)	4/60/80 (5)	4/10/10 (3)	0	0
2	洛夫林10 Lovrin 10	3/5/T (1)	3/5/10 (8)	4/40/60 (6)	4/10/T (4)	0	0
3	洛夫林13 Lovrin 13	0	3/20/40 (4)	4/20/60 (6)	4/20/T (4)	0	0
4	抗引655 Kangyin 655	0	3/10/20 (5)	4/20/40 (5)	0	0	0
5	中四（无芒）Zhong 4 (un-awn)	0	0	4/40/60 (5)	0	0	0
6	贵农22 Guinong 22	0	4/10/10 (2)	0	4/T/T (2)	0	0
7	Triticum spelta album	0	0	0	0	0	0
8	Hybrid 46	0	0	0	4/10/10 (3)	0	0
9	Reichersberg 42	0	0	4/10/10 (3)	4/10/T (3)	0	0
10	Heines Peko	0	3/5/10 (6)	4/60/80 (5)	4/10/20 (4)	0	0
11	Nord Desprez	0	0	4/80/80 (6)	4/10/T (3)	4/10/T (2)	0
12	Compair	0	3/5/10 (2)	4/40/40 (5)	4/10/10 (3)	2/T/T (1)	3/T/T (1)
13	Carsten V	0	3/5/T (3)	4/20/20 (6)	0	3/T/T (2)	0
14	Spaldings Prolific	0	3/10/T (4)	4/10/20 (4)	4/10/T (2)	0	0
15	Heines Ⅶ	0	0	4/10/20 (4)	0	0	0
16	Joss Cambier	0	0	0	0	0	0
17	Mega	0	0	0	3/5/T (2)	0	0
18	Hobbit	0	3/T/T (4)	4/20/40 (5)	3/10/T (4)	3/40/20 (4)	0
19	铭贤169 Mingxian 169	4/60/40 (1)	4/40/20 (7)	4/60/60 (4)	4/20/10 (3)	4/40/20 (2)	4/40/40 (3)
20	铭贤1696/Yr5 Mingxian 1696/Yr5	0	3/10/T (2)	4/60/60 (6)	0	0	0
21	铭贤1696/Yr10 Mingxian 1696/Yr10	0	4/10/10 (5)	4/80/80 (4)	4/10/10 (6)	4/60/60 (7)	0
22	周麦22 Zhoumai 22	0	0	0	0	4/10/10 (4)	0
23	烟农15 Yannong 15	0	3/5/T (5)	4/80/80 (2)	4/20/20 (4)	0	0
24	鲁麦23 Lumai 23	3/10/10 (4)	4/20/10 (2)	4/80/80 (4)	4/60/40 (6)	0	0
25	冀麦38 Jimai 38	0	4/10/20 (6)	4/20/80 (6)	4/20/10 (6)	0	0
26	石4185 Shi 4185	0	3/10/10 (3)	4/20/20 (5)	4/60/30 (3)	4/T/T (3)	0
27	晋太170 Jintai 170	0	4/20/20 (1)	4/60/40 (3)	4/20/40 (4)	4/10/20 (3)	0
28	杨麦158 Yangmai 158	0	3/5/T (2)	4/20/20 (5)	0	0	0
29	兰天19 Lantain 19	3/5/T (4)	0	0	0	0	0
30	兰天31 Lantian 31	3/5/T (1)	0	4/T/T (2)	0	0	0
31	兰天26 Lantian 26	3/10/T (4)	0	0	0	0	0
32	天选50 Tianxuan 50	3/5/T (1)	0	0	0	0	0
33	陇鉴386 Longjian 386	0	0	4/T/T (2)	0	4/40/60 (7)	0
34	Chancellor	0	4/20/20 (1)	0	4/20/20 (4)	4/30/40 (7)	0
35	Ulka/8cc	0	4/20/30 (5)	0	3/5/T (1)	—	0
36	Maris Huntsman	0	—	4/10/10 (2)	—	0	0
37	小白冬麦 Xiaobaidongmai	0	3/10/T (1)	0	4/40/10 (2)	0	0
38	Khaplic/8cc	0	3/10/10 (1)	0	4/10/10 (5)	0	0
39	Armada	0	0	0	0	0	0
40	81-7241	0	3/20/20 (4)	4/10/T (4)	4/80/40 (3)	—	0
41	Aquila	0	—	4/5/10 (2)	—	4/20/20 (3)	—
42	赤牙糙 Chiyacao	0	4/10/30 (4)	4/10/10 (2)	4/80/60 (4)	0	0
43	蚂蚱麦 Mazhamai	0	3/10/T (3)	4/10/T (3)	4/5/10 (3)	0	—
44	92R137	0	0	4/10/10 (5)	4/T/T (2)	0	0
45	保丰104 Baofen 104	0	3/10/T (3)	4/10/T (1)	4/40/40 (5)	0	4/40/20 (5)
46	高加索 Gaojiasuo	0	0	4/20/10 (4)	0	0	0
47	京双16 Jingshuang 16	0	3/20/10 (1)	4/10/10 (1)	0	0	0
48	白兔3号 Baitu No. 3	0	3/10/20 (5)	4/10/10 (3)	4/10/10 (2)	0	—
49	绵麦37 Mianmai 37	0	0	4/10/10 (3)	0	0	0
50	繁6 Fan 6	0	4/5/T (3)	4/T/T (3)	4/10/T (3)	0	0
51	绵阳11 Mianyang 11	0	3/40/40 (5)	4/T/T (3)	4/10/T (3)	0	0
52	川麦107 Chuanmai 107	3/10/T(2)	2/5/T (2)	4/10/T (1)	4/T/T (1)	0	0
53	川农19 Chuannong 19	3/5/T(4)	0	4/20/T (1)	0	0	0
54	川农27 Chuannong 27	3/5/T(2)	0	4/20/10 (4)	0	0	0
55	川麦58 Chuanmai 58	3/5/T(1)	0	0	4/T/T (2)	0	0
56	川育23 Chuanyu 23	0	0	0	0	0	0
57	Avocet S*6/Yr9	0	0	4/5/10 (2)	0	0	0
58	Avocet S*6/Yr15	0	0	4/10/10 (2)	—	0	0
59	Avocet S*6/Yr24	0	0	0	0	0	0
60	Avocet S*6/Yr26	0	0	0	0	0	0
61	Jupateco R Yr18	0	0	0	0	0	0
62	C591	0	0	4/10/10 (3)	0	0	0
63	TcLr9	0	0	0	4/10/T (4)	0	0
64	TcLr16	0	0	4/T/T (2)	4/40/10 (3)	0	0
65	TcLr19	0	4/40/10 (1)	4/10/10 (3)	0	0	0
66	TcLr24	0	3/10/10 (4)	4/10/10 (2)	0	0	0
67	TcLr26	0	3/5/T (4)	4/10/T (2)	0	0	0
68	TcLr38	0	0	0	0	0	0
69	TcLr46	0	0	4/60/40 (3)	0	0	0
70	ISr5 Ra	0	0	4/60/80 (4)	0	0	0
71	ISr6 Ra	0	0	0	0	0	0
72	Verstein Sr9e	0	3/40/20 (6)	0	—	0	0
73	W2691Sr10	0	4/10/10 (6)	4/10/20 (2)	4/20/40 (5)	0	0
74	ISr11 Ra	0	0	0	0	0	0
75	CnS-T-mono-deri=Sr21	0	3/T/T (3)	0	0	0	0
76	LcSr24Ag	0	3/20/T (5)	4/10/T (1)	4/10/10 (3)	0	0
77	Ealge Sr26	0	2/5/T (1)	4/10/10 (1)	4/5/T (1)	0	0
78	BtSr30 Wst	0	3/10/T (2)	0	0	0	0
79	Sr31/6*LMPG	3/5/T (1)	2/T/T (2)	4/5/T (1)	4/5/T (3)	0	0
80	W2691 SrTt-1=Sr36	0	3/10/T (5)	4/5/T (1)	4/5/T (2)	0	0
81	Trident Sr38	0	0	0	0	0	0
82	Little Club	0	4/20/30 (1)	0	0	0	4/T/T (2)

Table 1

Table 2

Table 3

Table 4

Table 5

Fig. 1

Fig. 2

Table 6

Fig. 3

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引物名称 Primer name	重复单元 Repeat motif		引物序列 Sequence (5′-3′)	染料 Dye	退火温度 Tm (℃)	参考文献 Reference
RJ03	(TGG)₈	F:	GCAGCACTGGCAGGTGG	FAM	61	文献[15] Reference [15]
		R:	GATGAATCAGGATGGCTCC			文献[15] Reference [15]
RJ04	(TGG)₈	F:	GTGGGTTGGGCTGGAGTC	HEX	58	文献[15] Reference [15]
		R:	GCTAATCCATTCCACGCACC			文献[15] Reference [15]
RJ12	(AC)₇	F:	ATC ATT CCG ATT TCT TTC TCA CC	FAM	56	文献[15] Reference [15]
		R:	TCA CAC TGA TCC CAA TAG ATC AG		56	文献[15] Reference [15]
RJ15	(TG)₇	F:	ATC GAG CAC GTC CAA ATC G	HEX	56	文献[15] Reference [15]
		R:	CAC TGG ACA GAC GAC GGT TG		56	文献[15] Reference [15]
RJ18	(TGT)₅	F:	CTG CCC ATG CTC TTC GTC	ROX	59	文献[15] Reference [15]
		R:	GAT GAA GTG GGT GCT GCT G		59	文献[15] Reference [15]
RJ20	(CAG)₄	F:	AGA AGA TCG ACG CAC CCG	ROX	56	文献[15] Reference [15]
		R:	CCT CCG ATT GGC TTA GGC		56	文献[15] Reference [15]
RJ21	(GTT)₆	F:	TTC CTG GAT TGA ATT CGT CG	FAM	55	文献[15] Reference [15]
		R:	CAG TTC TCA CTC GGA CCC AG		55	文献[15] Reference [15]
RJ24	(GTT)₅₊₉	F:	TTG CTG AGT AGT TTG CGG TGA G	HEX	58	文献[15] Reference [15]
		R:	CTC AAG CCC ATC CTC CAA CC			文献[15] Reference [15]
RJ27	(TC)₁₀	F:	CGTCCCGACTAATCTGGTCC	ROX	52	文献[15] Reference [15]
		R:	ATGAGTTAGTTTAGATCAGGTCGAC			文献[15] Reference [15]
CPS08	(CAG)₁₄	F:	GAT AAG AAA CAA GGG ACA GC	FAX	56	文献[2] Reference [2]
		R:	CAG TGA ACC CAA TTA CTC AG			文献[2] Reference [2]
CPS09	(GTT)₉	F:	CGG GAG AAG ACC TGA GC	ROX	55	文献[2] Reference [2]
		R:	AGA AAA CGG AAT GTA ATG TG			文献[2] Reference [2]
CPS10	(TAG)₈	F:	TCT ACT GGG CAG ACT GGT C	HEX	56	文献[2] Reference [2]
		R:	CGG TTT GTT TTG TCG TTT C			文献[2] Reference [2]
CPS15	(ATG)₅	F:	GAT GGG GAA AAG TAA GAA GT	FAM	56	文献[2]
		R:	GGT GGG GGA TGT AAG TAT GTA			Reference [2]
CPS34	(TTTGG)₄	F:	GTT GGC TAC GAG TGG TCA TC	HEX	56	文献[2]
		R:	TAA CAC TAC ACA AAA GGG GTC			Reference [2]
CPS36	(CTCTAG)₃	F:	TCC AGG CAG TAA ATC AGA CGC	ROX	58	文献[2]
		R:	ATC AGC AGG TGT AGC CCC ATC			Reference [2]

引物 Primer name	等位基因 Allele	均匀度 Evenness	缺失比率 Missing ratio (%)
RJ12	6	0.3695	2.72
RJ15	6	0.3383	5.63
RJ18	12	0.7109	8.89
RJ27	4	0.2719	4.72
RJ20	5	0.8056	8.71
RJ21	6	0.5285	2.36
RJ24	7	0.8571	3.63
CPS08	6	0.7591	0.36
CPS09	4	0.8648	9.62
CPS10	2	0.2997	0
CPS15	7	0.8171	1.09
CPS34	2	0.5949	0
CPS36	3	0.6670	0.36
RJ03	7	0.8327	5.63
RJ04	4	0.2982	6.17
平均Mean	5.4	0.6010	3.99

群体 Population	样本数 N	多基因座基因型 MLG	重采基因型编号及数量 Resampled MLG codes and numbers	基因型多样性 G	标准化关联指数rbarD	P值 P value
文县 Wenxian	27	23	61 (2), 427 (3), 429 (2)	0.852	0.0139	0.186
临洮 Lintao	132	111	29 (3), 74 (2), 169 (2), 184 (6), 185 (4), 187 (2), 192 (2), 193 (2), 194 (3), 197 (3), 210 (2), 228 (2)	0.841	0.1163	0.001
平凉 Pingliang	156	152	94 (2), 251 (2), 260 (2), 433 (2)	0.974	0.0286	0.001
临夏 Linxia	178	164	141 (5), 143 (2), 146 (2), 254 (2), 289 (2), 308 (2), 314 (2), 320 (2), 394 (2), 489 (2)	0.921	0.0110	0.010
互助 Huzhu	47	45	358 (2), 358 (2)	0.957	0.0416	0.002
西宁 Xining	11	10	272 (2)	0.909	0.0718	0.025
总数 Total	551	505	32	0.917	0.0260	0.001

	文县Wenxian	临洮Lintao	平凉Pingliang	临夏Linxia	互助Huzhu	西宁Xining
文县 Wenxian		2.408	2.337	3.408	2.916	1.074
临洮 Lintao	0.143^***		3.567	3.541	4.426	1.036
平凉 Pingliang	0.156^***	0.098^***		6.238	5.217	2.601
临夏 Linxia	0.114^***	0.103^***	0.056^***		8.055	2.072
互助 Huzhu	0.112^***	0.084^***	0.069^***	0.043^***		1.373
西宁 Xining	0.234^***	0.246^***	0.124^***	0.138^***	0.178^***

变异来源 Source of variance	自由度 Degree of freedom	变异组分 Variance component	变异百分数 Percentage of variation (%)	遗传分化系数 F_ST	P值 P value (rand≥data)
群体间Among populations	5	0.393	13	0.131	0.001
样本间Among individual	545	0.620	21
样本内Within individual	551	1.983	66
总数Total	1101	2.996	100

Surveillance and Genetic Diversity Analysis of Puccinia striiformis f. sp. tritici in Gansu and Qinghai Provinces

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