大豆芽期拟茎点种腐病鉴定体系构建及抗性种质筛选

doi:10.3864/j.issn.0578-1752.2024.11.004

Abstract

Abstract:

【Objective】An accurate and rapid indoor evaluation system was established by using soybeans with different resistance levels to Phomopsis seed decay as test materials. And then 170 soybean germplasm accessions were employed to screened out disease-resistant varieties, so as to provide methods and material basis for high-throughput assessment of Phomopsis seed decay in soybean and cultivation of resistant varieties.【Method】In terms of establishing a reliable evaluation method for Phomopsis seed decay, Qihuang 34, Williams, Zhongzuo 09-560, z13-631-2, ZDD26268, Chenxiqingpidou 1 and Tongxianhuangdou were selected as experimental materials. For each soybean accession, the seeds with uniform size and undamaged seed coat were germinated in the dark after disinfection. At different germination stages, the pathogen of Phomopsis seed decay was inoculated for 24 h, 48 h, 72 h and 96 h. The mycelium coverage rate and seed decay rate of seed surface under different infection time were counted to determine the optimal identification period for evaluating Phomopsis seed decay in soybean. Then, the resistance of 170 different soybean germplasms in natural population was identified by using the coverage rate of mycelium on the surface of seeds and the decay rate of seeds as evaluation indexes. The high disease resistance varieties were screened based on 5 disease resistance levels.【Result】The soybean accessions showed the most significant differences in disease resistance levels after 96 h of germination when mycelium coverage rate and seed decay rate of soybean surface were used as evaluation indexes. Further comparison of the incidence of 24 h, 48 h, 72 h and 96 h after infection showed that the difference in disease resistance between different varieties after infection for 72 h was the most obvious. Therefore, it was the most suitable period, 72 h of infection at the bud stage after 96 h of germination, for evaluating the resistance level of different soybean varieties to Phomopsis seed decay. The resistance of 170 soybean varieties to Phomopsis seed decay was identified and classified into five disease resistance grades, namely, high resistance, medium resistance, medium susceptibility, susceptibility and high susceptibility. Among them, there were 30 varieties of grade I (high resistance to disease), 51 varieties of grade Ⅱ (medium resistance to disease), 71 varieties of grade Ⅲ (medium disease susceptibility), 4 varieties of grade Ⅳ (disease susceptibility) and 14 varieties of grade V (high disease susceptibility), idicating that there are extensive variations in the resistance to Phomopsis seed decay of soybean germplasm resources in China.【Conclusion】In this study, the most optimum stage of disease identification was considered as soybean seeds after 96 h germination to infect the Phomopsis longicolla for 72 h. After that, the mycelium coverage rate and seed decay rate of soybean surface were counted as evaluation parameters. The evaluation system has high accuracy and reliability, which can provide an effective method for high-throughput identification of different varieties in the laboratory. And 30 highly resistant varieties were further screened to provide a material basis for the breeding of resistant varieties.

Key words: soybean, Phomopsis seed decay, evaluation system, disease-resistant variety

MIAO Long, YANG Lei, XU JingHao, LI Na, WANG FeiYu, QIU LiJuan, WANG XiaoBo. Establishment of Evaluation System and Screening of Disease- Resistant Accessions for Phomopsis Seed Decay in Soybean Germination Stage[J].Scientia Agricultura Sinica, 2024, 57(11): 2092-2101.

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Table 1

Table 2

Table 3

Table 4

30 soybean varieties with high resistance to Phomopsis seed decay"

品种名称 Name of varieties	菌丝覆盖率平均值±标准差 Mean±SD (%)	种子腐烂率平均值±标准差 Mean±SD (%)	抗性等级 Grade
铁荚子Tiejiazi	0.00±0.00	0.00±0.00	Ⅰ级（高抗）HR
金元1 Jinyuan No.1	0.00±0.00	0.00±0.00	Ⅰ级（高抗）HR
陈茬大豆Chenchadadou	0.00±0.00	0.00±0.00	Ⅰ级（高抗）HR
八月炸Bayuezha	0.00±0.00	0.00±0.00	Ⅰ级（高抗）HR
泉变11 Quanbian 11	10.33±2.52	0.00±0.00	Ⅰ级（高抗）HR
中黄30 Zhonghuang 30	7.00±2.00	0.00±0.00	Ⅰ级（高抗）HR
黑豆Heidou	31.33±3.21	0.00±0.00	Ⅰ级（高抗）HR
大屯小黑豆Datunxiaoheidou	40.67±4.04	7.33±2.52	Ⅰ级（高抗）HR
WDD02069	0.00±0.00	11.67±2.08	Ⅰ级（高抗）HR
青皮豆Qingpidou	0.00±0.00	8.33±1.53	Ⅰ级（高抗）HR
平顶黄豆Pingdinghuangdou	0.00±0.00	10.00±2.00	Ⅰ级（高抗）HR
茶豆Chadou	0.00±0.00	13.00±3.00	Ⅰ级（高抗）HR
涟源虎皮豆Lianyuanhupidou	0.00±0.00	12.67±2.08	Ⅰ级（高抗）HR
冬大豆Dongdadou	31.33±3.51	14.67±2.52	Ⅰ级（高抗）HR
吉青3号Jiqing No.3	39.67±2.52	21.00±3.61	Ⅰ级（高抗）HR
红豆Hongdou	80.33±2.52	22.00±2.65	Ⅰ级（高抗）HR
房山黑豆-1 Fangshanheidou -1	81.67±2.08	22.33±2.52	Ⅰ级（高抗) HR
青皮1 Qingpi1	0.00±0.00	25.67±3.06	Ⅰ级（高抗）HR
黑壳豆Heikedou	3.33±2.08	26.67±2.08	Ⅰ级（高抗）HR
陇中黄602 Longzhonghuang 602	30.67±4.04	22.67±3.21	Ⅰ级（高抗）HR
AGS162	51.00±2.65	22.33±3.06	Ⅰ级（高抗）HR
迟黄豆1 Chihuangdou 1	53.67±3.21	24.67±3.51	Ⅰ级（高抗）HR
中作09-560 Zhongzuo 09-560	75.33±4.73	27.00±2.00	Ⅰ级（高抗）HR
WDD01831	0.00±0.00	30.00±4.00	Ⅰ级（高抗）HR
ZDD26268	0.00±0.00	32.67±3.06	Ⅰ级（高抗）HR
浙鲜豆2号Zhexiandou No.2	7.67±2.08	30.67±4.04	Ⅰ级（高抗）HR
新垒头茶豆Xinleitouchadou	20.00±3.00	33.33±3.06	Ⅰ级（高抗）HR
PI438342	29.67±1.53	32.33±2.08	Ⅰ级（高抗）HR
中江红黄豆Zhongjianghonghuangdou	73.67±4.04	32.00±1.73	Ⅰ级（高抗）HR
山宁7号Shanning 7	81.33±4.16	29.33±3.06	Ⅰ级（高抗）HR

Table 4

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品种名称 Name of varieties	萌发时长 Germination time (h)
	0		48		72		96
	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率 Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)
Williams	64.0±13.4b	14.0±5.5b	50.0±15.8b	12.0±8.4b	68.0±14.8ab	70.0±7.1a	90.0±11.5a	72.5±9.6a
中作09-560 Zhongzuo 09-560	55.0±1.3ab	12.5±5.0b	46.0±8.90b	28.0±4.5ab	66.7±5.8a	25.0±5.7a	58.0±8.4ab	26.0±5.5ab
z13-631-2	82.50±9.6a	67.5±15.0a	46.7±15.3b	60.0±5.0ab	13.3±5.7c	53.3±5.8ab	60.0±8.9b	50.0±6.3b
ZDD26268	72.0±11.7a	21.7±14.7b	55.0±7.1ab	55.0±7.1a	43.3±20.8b	60.0±10.0a	12.0±4.5c	20.0±7.1b
辰溪青皮豆1 Chenxiqingpidou 1	72.5±9.6ab	57.5±9.6b	80.0±10.0a	74.0±8.9a	60.0±10.0b	63.3±5.8ab	60.0±8.2b	55.0±5.8b
通县黄豆 Tongxian huangdou	55.0±5.8a	15.0±5.8c	66.7±25.2a	73.3±11.5ab	56.0±15.20a	56.0±11.4a	45.0±10.0a	25.0±5.8bc
齐黄34 Qihuang 34	12.5±10.0a	10.0±5.8b	11.7±4.1a	15.0±5.5b	8.3±4.1a	15.0±5.5b	13.3±5.8a	41.7±9.6a

品种名称 Name of varieties	侵染时长 Infection time (h)
	24		48		72		96
	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)	菌丝覆盖率Coverage rate (%)	种子腐烂率Decay rate (%)
Williams	0.00c	0.00c	40.00±8.40b	40.00±5.50b	90.00±11.50a	72.50±9.60a	91.67±9.83a	83.30±7.53a
中作09-560 Zhongzuo 09-560	0.00c	0.00c	0.00c	0.00c	58.00±8.40b	26.00±5.50b	84.00±8.90a	50.00±12.20a
z13-631-2	0.00d	0.00d	18.00±4.70c	16.00±5.50c	60.00±8.94b	50.00±6.32b	95.00±5.48a	93.30±8.37a
ZDD26268	0.00c	16.00±5.50b	0.00c	16.70±5.20b	12.00±4.50b	20.00±7.10b	30.00±8.20a	35.00±5.80a
辰溪青皮豆1 Chenxiqingpidou 1	17.50±9.60b	0.00d	27.50±9.60b	17.50±9.60c	60.00±8.20a	55.00±5.80b	67.50±9.60a	70.00±14.10a
通县黄豆 Tongxian huangdou	0.00b	0.00c	0.00b	0.00c	45.00±10.00a	25.00±5.80b	47.50±9.60a	35.00±17.30a
齐黄34 Qihuang 34	0.00b	0.00c	0.00b	22.00±4.50b	13.30±5.80a	41.70±9.60a	0.00b	52.50±15.00a

抗性等级 Grade	品种数量Number	菌丝覆盖率Coverage rate (%)	种子腐烂率 Decay rate (%)	占比 Fraction (%)	菌丝覆盖率平均值±标准差 Mean±SD (%)	菌丝覆盖率变异系数 CV of coverage rate (%)	腐烂率平均值± 标准差 Mean±SD (%)	腐烂率变异系数 CV of decay rate (%)
Ⅰ级（高抗）HR	30	<80	0-30	17.65	24.00±28.40	118.30	17.00 ±12.00	70.59
Ⅱ级（中抗）MR	51	<80	31-60	30.00	24.51±20.98	85.60	49.80±9.22	18.51
Ⅲ级（中感）MS	71	<80	61-100	41.76	34.07±29.61	86.91	79.55±17.00	21.37
Ⅲ级（中感）MS	71	≥80	0-30	41.76	34.07±29.61	86.91	79.55±17.00	21.37
Ⅳ级（感病）S	4	≥80	31-60	2.35	96.25±4.78	4.97	54.00±4.78	8.85
Ⅴ级（高感）HS	14	≥80	61-100	8.23	93.00±7.00	7.53	89.64±13.65	15.23

Establishment of Evaluation System and Screening of Disease- Resistant Accessions for Phomopsis Seed Decay in Soybean Germination Stage

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