Simple and rapid detection of Tilletia horrida causing rice kernel smut in rice seeds.
A simple and rapid method for the detection of Tilletia horrida, the causal agent of rice kernel smut, in rice seeds is developed based on specific polymerase chain reaction (PCR). To design the specific primers for the detection of T. horrida, partial sequences of internal transcribed spacer (ITS) DNA region of T. horrida, T. controversa, T. walkeri, T. ehrhartae, T. indica and T. caries were analyzed and compared. A 503-bp fragment was amplified with the designed primers from the T. horrida genomic DNA. However, no PCR product was obtained from the DNA of other five Tilletia species and 22 fungal plant pathogens tested in the present work indicating the specificity of the primers for the detection of T. horrida. The PCR was performed by directly using the spores, isolated from the 21 different rice seed samples, as template DNA. The T. horrida was detected in 6 of the samples, indicating that 28.6% of the rice samples were contaminated with the kernel smut pathogen. This simple PCR based diagnostic assay can be applied for the direct and rapid detection and identification of T. horrida to screen large numbers of rice seed samples.
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Yu
Y
Chen
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Xue
X
Yang
Anhui Entry-Exit Inspection and Quarantine Bureau, Hefei 230022, China.
Jian
J
Yao
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.|Department of Biotechnology, Mandalay Technological University, Mandalay, Republic of the Union of Myanmar.
Ei Phyu
EP
Kyaw
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Ai-Fang
AF
Zhang
Anhui Entry-Exit Inspection and Quarantine Bureau, Hefei 230022, China.
Yun-Fei
YF
Li
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Chun-Yan
CY
Gu
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Hao-Yu
HY
Zang
Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China.|Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, China.|Laboratory of Quality &Safety Risk Assessment for Agro-Products (Hefei), Ministry of Agriculture, China.
Tong-Chun
TC
Gao
D001487:Basidiomycota / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D012275:Oryza / Q000235:genetics / Q000254:growth & development / Q000382:microbiology*; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics / Q000382:microbiology
D004271:DNA, Fungal