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印度卡纳塔克邦种植的谷物上出现的有毒和植物致病镰刀菌物种的多样性。

Diversity of toxic and phytopathogenic Fusarium species occurring on cereals grown in Karnataka state, India.

作者信息

Nagaraja H, Chennappa G, Poorna Chandra Rao K, Mahadev Prasad G, Sreenivasa M Y

机构信息

Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysore, 570 006, Karnataka, India.

出版信息

3 Biotech. 2016 Jun;6(1):57. doi: 10.1007/s13205-016-0399-5. Epub 2016 Feb 13.

Abstract

A total of 198 cereal samples (53 maize, 54 sorghum, 37 paddy and 54 wheat) were collected from 11 districts of Karnataka to understand the percent infection (PI), relative density (RD) and their frequency (Fr) caused by Fusarium spp. All samples were screened by agar plating method using MGA 2.5 agar media and incubated at 25 ± 2 °C for 3-5 days. The study revealed the association of 10 different Fusarium species known trichothecene producers such as Fusarium acuminatum, F. avenaceum, F. crookwellense, F. culmorum, F. equiseti, F. graminearum, F. nivale, F. poae, F. sambucinum and F. sporotrichioides along with non-trichothecene producers like F. anthophilum, F. oxysporaum, F. proliferatum, F. semitectum, F. solani, and F. verticillioides. All the ten isolated potential trichothecene producing Fusarium species were analyzed for their ability to produce trichothecenes by using thin layer chromatography method. The highest infection of Fusarium spp. in maize was by F. verticillioides with PI of (2.95 %), with RD of (15.16 %) and highest Fr was by F. graminearum (79.24 %) and the lowest was F. avenasium with PI of (0.13 %). For sorghum maximum PI was by F. verticillioides (3.02 %), with F. graminearum having highest RD (14.39 %) and with F. verticillioides highest Fr. (72.22 %). In paddy highest PI was by F. verticillioides (3.21 %) and the least was by F. avenaceum (0.09 %). Similarly in wheat the highest PI was by F. verticillioides (2.76 %) while lowest was by F. avenaceum (0.10 %). The highest Fr was with F. graminearum (79.62 %) while the lowest was by F. avenaceum (3.70 %) and the highest RD was by F. graminearum (22.04 %) and lowest was by F. solani (0.72 %). The manually identified Fusarium spp. were further confirmed by PCR-based detection using ITS1 and ITS4 primers followed by sequencing of the PCR amplicons. PCR studies confirmed that all the tested fungal isolates belongs to Fusarium spp. with the amplicon size of 600 bp. Sequencing and the blast data from NCBI data base confirmed the sequence similarity of 99 % to the genus Fusarium and accession numbers were obtained. Chemotyping studies showed that the isolated Fusarium species are known to produce different types of trichothecenes. The study revealed the diversity in phytopathogenic Fusarium spp. in major cereal crops growing in different agro-climatic regions of Karnataka, India.

摘要

从卡纳塔克邦的11个地区共采集了198份谷物样本(53份玉米、54份高粱、37份稻谷和54份小麦),以了解镰刀菌属引起的感染率(PI)、相对密度(RD)及其频率(Fr)。所有样本均采用MGA 2.5琼脂培养基通过琼脂平板法进行筛选,并在25±2℃下培养3 - 5天。该研究揭示了10种不同的已知产单端孢霉烯族毒素的镰刀菌物种的关联,如锐顶镰刀菌、燕麦镰刀菌、克威尔镰刀菌、禾谷镰刀菌、木贼镰刀菌、禾谷镰刀菌、雪腐镰刀菌、早熟禾镰刀菌、接骨木镰刀菌和拟枝孢镰刀菌,以及非产单端孢霉烯族毒素的物种,如嗜花镰刀菌、尖孢镰刀菌、轮枝镰刀菌、半裸镰刀菌、茄病镰刀菌和串珠镰刀菌。使用薄层色谱法分析了所有十种分离出的潜在产单端孢霉烯族毒素的镰刀菌物种产生单端孢霉烯族毒素的能力。玉米中镰刀菌属的最高感染率是由轮枝镰刀菌引起的,感染率为(2.95%),相对密度为(15.16%),频率最高的是禾谷镰刀菌(79.24%),最低的是燕麦镰刀菌,感染率为(0.13%)。高粱的最高感染率是由轮枝镰刀菌(3.02%)引起的,禾谷镰刀菌的相对密度最高(14.39%),轮枝镰刀菌的频率最高(72.22%)。稻谷中最高感染率是由轮枝镰刀菌(3.21%)引起的,最低的是燕麦镰刀菌(0.09%)。同样,小麦中最高感染率是由轮枝镰刀菌(2.76%)引起的,最低的是燕麦镰刀菌(0.10%)。频率最高的是禾谷镰刀菌(79.62%),最低的是燕麦镰刀菌(3.70%),相对密度最高的是禾谷镰刀菌(22.04%),最低的是茄病镰刀菌(0.72%)。通过使用ITS1和ITS4引物进行基于PCR的检测,随后对PCR扩增产物进行测序,进一步确认了人工鉴定的镰刀菌属物种。PCR研究证实,所有测试的真菌分离株均属于镰刀菌属,扩增子大小为600 bp。测序和来自NCBI数据库的比对数据证实与镰刀菌属的序列相似性为99%,并获得了登录号。化学分型研究表明,分离出的镰刀菌物种已知会产生不同类型的单端孢霉烯族毒素。该研究揭示了印度卡纳塔克邦不同农业气候区主要谷类作物中植物病原镰刀菌属的多样性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ab/4752948/1eb943cdd3af/13205_2016_399_Fig1_HTML.jpg

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