Please wait a minute...
浙江大学学报(农业与生命科学版)  2018, Vol. 44 Issue (1): 41-48    DOI: 10.3785/j.issn.1008-9209.2017.03.121
植物保护     
向日葵黄萎病种子带菌研究
张贵1,张园园1,田永伟1,赵晓军1,张光2,周洪友1,景岚1,赵君1*
1.内蒙古农业大学农学院,呼和浩特 010018;2.内蒙古乌拉特前旗农业技术推广中心,内蒙古 乌拉特前旗 014400
Detection of contamination of sunflower seed coats with Verticillium dahliae
ZHANG Gui1, ZHANG Yuanyuan1, TIAN Yongwei1, ZHAO Xiaojun1, ZHANG Guang2, ZHOU Hongyou1, JING Lan1, ZHAO Jun1*
1. College of Agronomy, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. Agricultural Technique Center of Wulateqianqi, Wulateqianqi 014400, Inner Mongolia, China
 全文: PDF(1533 KB)   HTML (
摘要:

为探究向日葵种子是否能够携带黄萎病菌进行远距离传播,以随机挑选的7个向日葵品种为研究材料,以种皮DNA 为模板,利用真菌通用引物ITS1/ITS4和ITS区内侧的黄萎菌特异引物进行巢氏聚合酶链式反应(polymerase chain reaction, PCR),将所获得的PCR条带测序后在GenBank 中进行比对。结果表明,供试的向日葵品种的种皮中所扩增出的PCR条带均来自黄萎病菌(Verticillium dahliae)。为了进一步确定上述检测结果,另挑选9 个不同向日葵品种的种子,对其种皮带菌率进行测定。结果表明,供试的9 个向日葵品种的种皮带菌率介于10.0%~25.0%之间,其中田间采集的3638C新鲜种子带菌率最低,仅为10.0%,而实验室留存的赤029X115R种子带菌率最高,为25.0%。为了进一步验证PCR 实验结果的准确性,利用表达绿色荧光蛋白(green fluorescentprotein, GFP)的黄萎病菌株接种向日葵植株根部,在荧光显微镜下对种皮进行观察。结果表明,在接种向日葵花盘上收获的种子的种皮上可以观察到GFP荧光信号。上述研究结果表明,向日葵种子的种皮可以携带黄萎病菌,而巢氏PCR可以作为一种检测向日葵黄萎病种子是否带菌的快速而可行的方法。

关键词: 巢氏聚合酶链式反应快速检测体系向日葵种子带菌黄萎病菌    
Abstract:

Sunflower Verticillium Wilt is a soil-borne disease caused by Verticillium dahliae, which can infect a variety of crops, resulting in a serious decline in both yield and quality. The conidia and microsclerotia in sunflower debris are the principal sources of infection. A nested polymerase chain reaction (PCR) was used to detect the contamination of V. dahliae in sunflower seed coats. Seven sunflower varieties were selected to isolate DNA from their seed coats, and then performed a nested PCR by using specific primers of V. dahliae and the universal primers of fungi (ITS1/ITS4). The PCR products were sequenced then BLAST-searched in GenBank. In order to confirm the PCR results, green fluorescent protein (GFP) labeled V. dahliae was used to inoculate sunflower roots. The seed coats were then removed from inoculated sunflower seeds and detected GFP signal under a fluorescence microscope. The results showed that all the amplified PCR bands belonged to V. dahliae isolated from the seed coats of the tested sunflower cultivars. Meanwhile, variable seed contamination ratios were found among different sunflower varieties. The lowest and highest contamination ratios were observed in 3638C (10.0%) and Chi029X115R (25.0%) varieties, respectively. Moreover, the GFP signals obtained on the seed coats of inoculated sunflowers further supported the PCR results. Based on the above results, we confirm that the sunflower seed coat is an important tissue for V. dahliae colonization, and it is the main carrier for the long distance transmission of the sunflower Verticillium Wilt. Additionally, we conclude that the nested PCR method is fast and highly reliable to detect V. dahliae contamination in sunflower seed coats.

Key words: seed-borne    nested polymerase chain reaction    Verticillium dahliae    sunflower    fast detection system
收稿日期: 2017-03-12 出版日期: 2017-05-09
CLC:  S 435.655  
基金资助: 国家特色油料产业技术体系项目(CARS-14);国家公益性行业(农业)科研专项(201503109)
通讯作者: 赵君(http://orcid.org/0000-0001-7207-9289)     E-mail: zhaojun@imau.edu.cn
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
张贵
田永伟
周洪友
赵君
张园园
赵晓军
张光
景岚

引用本文:

张贵,张园园,田永伟,赵晓军,张光,周洪友,景岚,赵君. 向日葵黄萎病种子带菌研究[J]. 浙江大学学报(农业与生命科学版), 2018, 44(1): 41-48.

ZHANG Gui, ZHANG Yuanyuan, TIAN Yongwei, ZHAO Xiaojun, ZHANG Guang, ZHOU Hongyou, JING Lan, ZHAO Jun. Detection of contamination of sunflower seed coats with Verticillium dahliae. Journal of Zhejiang University (Agriculture and Life Sciences), 2018, 44(1): 41-48.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2017.03.121        http://www.zjujournals.com/agr/CN/Y2018/V44/I1/41

[1] 孙晓婷, 鹿秀云, 张敬泽, 祝水金. 浙江棉花黄萎病菌致病型菌株的鉴定及高温抑制病害的表征分析[J]. 浙江大学学报(农业与生命科学版), 2016, 42(6): 671-678.
[2] 赵亦静, 倪密, 诺林, 王学德*. 人工合成抗菌肽对棉花黄萎病菌的抑菌效果[J]. 浙江大学学报(农业与生命科学版), 2013, 39(1): 11-.