水稻离体叶片纹枯病接种样品菌量检测方法研究

doi:10.3969/j.issn.1004-1524.2018.10.11

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (10): 1686-1693.DOI: 10.3969/j.issn.1004-1524.2018.10.11

水稻离体叶片纹枯病接种样品菌量检测方法研究

曲海艳^{1, 2}, 袁正杰², 潘龙玉^{1, 2}, 何海燕², 许赵蒙^{1, 2}, 瞿绍洪^{2, *}

1.浙江师范大学化学与生命科学学院,浙江金华 321004;
2.浙江省农业科学院病毒学与生物技术研究所,浙江杭州 310021

收稿日期:2017-12-29 出版日期:2018-10-25 发布日期:2018-11-02
通讯作者: 瞿绍洪,E-mail:squ@mail.zaas.ac.cn
作者简介:曲海艳(1992—),女,山东烟台人,硕士研究生,主要从事水稻生物技术研究。E-mail:563561809@qq.com
基金资助:
国家自然科学基金(316720161005943); 浙江省农业科学院国际合作项目; 浙江省农业科学院扶持学科项目; 浙江省农业科学院省部共建国家重点实验室培育基地项目(2010DS700124-KF1210); 转基因生物新品种培育重大专项(2012ZX08009001)

Studies on quantification method of rice sheath blight pathogen (Rhizoctonia solani) in inoculated rice detached-leaf samples

QU Haiyan^{1, 2}, YUAN Zhengjie², PAN Longyu^{1, 2}, HE Haiyan², XU Zhaomeng^{1, 2}, QU Shaohong^{2, *}

1.College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004,China;
2.Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China

Received:2017-12-29 Online:2018-10-25 Published:2018-11-02

摘要/Abstract

摘要： 利用qPCR方法对水稻离体叶片纹枯病接种材料进行纹枯病菌含量的检测,从而对水稻发病程度进行定量分析。各水稻品种接种样品的纹枯病菌DNA含量,从低到高的顺序为:YSBR1、徐稻3号、台北309、泰粳394和Lemont。通过对接种实验条件的精确控制,离体叶片的病斑扫描,以及相对病斑面积的统计分析,验证了关于病害严重程度的qPCR检测结果。大田接种试验中,YSBR1、Lemont和泰粳394的平均病级呈显著差异;这进一步证明,通过纹枯病菌DNA的qPCR方法,能够快速有效地定量检测不同水稻品种的纹枯病抗性差异。该研究可为量化纹枯病发病程度、评价水稻品种的抗性水平及进行纹枯病的早期预报,提供有效的检测手段。

关键词: 水稻, 纹枯病, 离体叶片接种, 菌量检测

Abstract: In this study, qPCR assays of the rice sheath blight (SB) pathogen inoculated to detached rice leaf tissues were performed to evaluate the disease level. Rice cultivars with the relative content of pathogen DNA in inoculated samples from the lowest to the highest are as below: YSBR1, Xudao 3, Taipei 309, Taijing 394, Lemont. The disease severity of leaf tissues indicated by the qPCR results was verified by precise control of the experimental conditions in SB inoculation, digital scanning of disease lesions, and statistical analysis of lesion areas. The average disease indexes of YSBR1, Lemont and Taijing 394 were significantly different from each other in field SB inoculation, further demonstrating that qPCR can be used to quantitatively evaluate the SB resistance level of rice cultivars. This study provides an effective method for quantification of the SB severity, evaluation of the resistance level of rice cultivars, and the forecast of the SB disease.

Key words: rice (Oryza sativa), sheath blight, detached-leaf inoculation, pathogen quantification

中图分类号:

S435.111.4⁺2

曲海艳, 袁正杰, 潘龙玉, 何海燕, 许赵蒙, 瞿绍洪. 水稻离体叶片纹枯病接种样品菌量检测方法研究[J]. 浙江农业学报, 2018, 30(10): 1686-1693.

QU Haiyan, YUAN Zhengjie, PAN Longyu, HE Haiyan, XU Zhaomeng, QU Shaohong. Studies on quantification method of rice sheath blight pathogen (Rhizoctonia solani) in inoculated rice detached-leaf samples[J]. , 2018, 30(10): 1686-1693.

参考文献

[1] 孟庆忠,刘志恒,王鹤影,等.水稻纹枯病研究进展[J].沈阳农业大学学报,2001,32(5):376-381.
MENG Q Z, LIU Z H, WANG H Y, et al.Research progress in rice sheath blight[J]. Journal of Shenyang Agricultural University, 2001, 32(5): 376-381. (in Chinese with English abstract)
[2] 左示敏,张亚芳,陈宗祥, 等. 水稻抗纹枯病遗传育种研究进展[J].中国科学(生命科学),2010,40(11):1014-1023.
ZUO S M, ZHANG Y F, CHEN Z X, et al. Current progress on genetics and breeding in resistance to rice sheath blight[J]. Scientia Sinica(Vitae), 2010, 40:(11): 1014-1023. (in Chinese with English abstract)
[3] PINSON S R M, CAPDEVIELLE F M, OARD J H. Confirming QTLs and finding additional loci conditioning sheath blight resistance in rice using recombinant inbred lines[J]. Crop Science, 2005, 45(2): 503-510.
[4] 左示敏,张亚芳,殷跃军. 田间水稻纹枯病抗性鉴定体系的确立与完善[J].扬州大学学报,2006,27(4):57-61+111.
ZUO S M, ZHANG Y F,YIN Y J, et al. Establishment and improvement of inoculation technique and rating system in researching rice sheath blight resistance in field[J].Journal of Yangzhou University,2006,27(4):57-61+111. (in Chinese with English abstract)
[5] JIA Y, CORREA V F, MC C A, et al.Rapid determination of rice cultivar responses to the sheath blight pathogen Rhizoctonia solani using a micro-chamber screening method[J]. Plant Disease, 2007, 91(5): 485-489.
[6] PRASAD B, EIZENGA G C.Rice sheath blight disease resistance identified in Oryza spp. accessions[J].Plant Disease, 2008, 92(11): 1503-1509.
[7] 马晨燕,袁正杰,杨海河,等. 水稻离体叶片抗纹枯病接种方法的研究[J].浙江农业学报,2016,28(10): 1730-1737.
MA C Y, YUAN Z J, YANG H H, et al.Studies on the detached-leaf inoculation method for determination of rice resistance to sheath blight (Rhizoctonia solani)[J]. Acta Agriculturae Zhejiangensis, 2016, 28(10): 1730-1737. (in Chinese with English abstract)
[8] SAYLER R J, YANG Y N.Detection and quantification of Rhizoctonia solani AG-1 IA, the rice sheath blight pathogeon, in rice using real-time PCR[J]. Plant Disease, 2007, 91(12): 1663-1668.
[9] 陈夕军,王玲,左示敏,等.水稻纹枯病寄主—病原物互作鉴别品种与菌株的筛选[J].植物病理学报, 2009, 39(5): 514-520.
CHEN X J, WANG L, ZUO S M, et al.Screening of cultivars and isolates for identifying interaction between host and pathogen of rice sheath blight[J]. Acta Phytopathologica Sinica, 2009, 39(5): 514-520. (in Chinese with English abstract)
[10] 徐国娟,袁正杰,左示敏,等.水稻苗期纹枯病抗性鉴定微室接种技术的改良[J].中国水稻科学, 2015, 29(1): 97-105.
XU G J, YUAN Z J, ZUO S M, et al.Improvement of the micro-chamber inoculation method for determination of rice seedling resistance to sheath blight (Rhizoctonia solani)[J]. Chinese Journal of Rice Science, 2015, 29(1): 97-105.(in Chinese with English abstract)
[11] 彭丽丽. 外源基因GAFP提高水稻纹枯病抗性的效应研究[D].扬州:扬州大学,2013.
PENG L L.Study on resistance to sheath blight for transgenic GAFP rice lines [D].Yangzhou: Yangzhou University, 2013.(in Chinese with English abstract)
[12] 曹玉鑫. 几丁质识别受体与Xa21的嵌合基因对水稻真菌病害的抗性研究[D].海口:海南大学,2013.
CAO Y X.The research on resistance of chimeric gene composed of chitin receptor and Xa21 against rice fungus deseases [D].Haikou: Hainan University, 2013. (in Chinese with English abstract)
[13] 崔华威,杨艳丽,黎敬涛,等.一种基于Photoshop的叶片相对病斑面积快速测定方法[J].安徽农业科学, 2009, 37(22): 10760-10762.
CUI H W, YANG Y L, LI J T, et al.A faster method for measuring relative lesion area on leaves based on software Photoshop[J].Journal of Anhui Agricultural Sciences, 2009, 37(22): 10760-10762. (in Chinese with English abstract)
[14] 王子斌,左示敏,李刚,等.水稻成株期对纹枯病的抗性表现研究[J].吉林农业大学学报, 2011, 33(2): 144-150.
WANG Z B, ZUO S M, LI G, et al.Study of resistance of rice to sheath blight at adult plant stage[J]. Journal of Jilin Agricultural University, 2011, 33(2): 144-150. (in Chinese with English abstract)
[15] 潘学彪,陈宗祥,徐敬友,等.不同接种调查方法对抗水稻纹枯病遗传研究的影响[J].江苏农学院学报, 1997, 18(3): 27-32.
PAN X B, CHEN Z X, XU J Y, et al.The effects of different methods of inoculation and investigation on genetic research of resistance to rice sheath blight[J].Journal of Jiangsu Agricultural College, 1997, 18(3): 27-32. (in Chinese with English abstract)
[16] RUSH M C, HOFF B J, MELLRATH W O.A uniform disease rating system for rice disease in the United States[C]//Proceedings of the 16th Rice Technical Working Group, Lake Charles, Louisiana, USA, 1976: 64.

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水稻离体叶片纹枯病接种样品菌量检测方法研究

Studies on quantification method of rice sheath blight pathogen (Rhizoctonia solani) in inoculated rice detached-leaf samples

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