中国农业科学 ›› 2024, Vol. 57 ›› Issue (11): 2092-2101.doi: 10.3864/j.issn.0578-1752.2024.11.004

• 专题:大豆抗病性、产量与品质的相关性 • 上一篇    下一篇

大豆芽期拟茎点种腐病鉴定体系构建及抗性种质筛选

苗龙1(), 杨蕾1, 许竞好1, 李娜1, 王飞宇1, 邱丽娟2(), 王晓波1()   

  1. 1 安徽农业大学农学院,合肥 230036
    2 中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业农村部作物基因资源与种质创制重点实验室,北京 100081
  • 收稿日期:2023-09-10 接受日期:2023-10-23 出版日期:2024-06-01 发布日期:2024-06-07
  • 通信作者:
    邱丽娟,E-mail:
    王晓波,E-mail:
  • 联系方式: 苗龙,E-mail:lmiao5@163.com。
  • 基金资助:
    安徽省自然科学基金(2308085MC88); 安徽省重点研究与开发计划(202204c06020026); 安徽农业大学引进和稳定人才项目(rc312003)

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

MIAO Long1(), YANG Lei1, XU JingHao1, LI Na1, WANG FeiYu1, QIU LiJuan2(), WANG XiaoBo1()   

  1. 1 College of Agriculture, Anhui Agricultural University, Hefei 230036
    2 Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Crop Gene Resource and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs, Beijing 100081
  • Received:2023-09-10 Accepted:2023-10-23 Published:2024-06-01 Online:2024-06-07

摘要:

【目的】以具有拟茎点种腐病不同抗性水平的大豆为试材,建立准确快速的大豆芽期室内病害鉴定体系,并基于170份自然群体大豆筛选出高抗病性品种,为大豆拟茎点种腐病的快速高通量鉴定和抗性品种培育提供方法及材料基础。【方法】选取已报道拟茎点种腐病抗性品种齐黄34、感病品种Williams,以及中作09-560、z13-631-2、ZDD26268、辰溪青皮豆1和通县黄豆,挑选各品种大小均一、种皮完好的种子,经过消毒后于黑暗中发芽,在不同萌发时长后分别接种拟茎点种腐病病原菌24、48、72和96 h,统计不同侵染时长下种子表层的菌丝覆盖率与种子腐烂率,确定评价大豆拟茎点种腐病的最适鉴定时期。随后,以该时期的种子表层菌丝覆盖率和种子腐烂率作为评价指标,对自然群体170份不同大豆种质进行抗性鉴定,并划分5个抗病等级,筛选出高抗病性品种。【结果】以种子表层的菌丝覆盖率和种子腐烂率作为评价指标,发现不同抗性水平的大豆在萌发96 h后接种拟茎点种腐病病原菌表现出最为明显的表型差异;进一步比较侵染24、48、72和96 h后的发病情况,发现侵染72 h后的不同品种之间抗病差异最为明显,因此,确定萌发96 h后的芽期大豆侵染72 h为评价不同品种拟茎点种腐病抗性水平的最适时期,随后对170份大豆自然品种进行拟茎点种腐病抗性鉴定,并划为高抗、中抗、中感、感病、高感5个抗病等级,其中Ⅰ级(高抗)品种30个、Ⅱ级(中抗)品种51个、Ⅲ级(中感)品种71个、Ⅳ级(感病)品种4个、Ⅴ级(高感)品种14个,表明大豆种质资源对拟茎点种腐病抗性存在广泛变异。【结论】以萌发96 h后的芽期大豆侵染病原菌72 h作为最佳鉴定时期、种子表层的菌丝覆盖率和种子腐烂率作为拟茎点种腐病的评价指标,该鉴定体系具有较高的准确性与可靠性,可为不同品种室内快速高通量鉴定提供有效方法,进一步筛选出30个高抗种质可为抗病品种选育提供材料基础。

关键词: 大豆, 拟茎点种腐病, 鉴定体系, 抗病种质

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