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A Nematode Calreticulin, Rs-CRT, Is a Key Effector in Reproduction and Pathogenicity of Radopholus similis.

Li Y, Wang K, Xie H, Wang YT, Wang DW, Xu CL, Huang X, Wang DS - PLoS ONE (2015)

Bottom Line: The reproductive capability and pathogenicity of R. similis were significantly reduced after treatment with Rs-crt dsRNA for 36 h.Using plant-mediated RNAi, we confirmed that Rs-crt expression was significantly inhibited in the nematodes, and resistance to R. similis was significantly improved in transgenic tomato plants.Plant-mediated RNAi-induced silencing of Rs-crt could be effectively transmitted to the F2 generation of R. similis; however, the silencing effect of Rs-crt induced by in vitro RNAi was no longer detectable in F1 and F2 nematodes.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Plant Nematology and Research Center of Nematodes of Plant Quarantine, Department of Plant Pathology, South China Agricultural University, Guangzhou, Guangdong, 510642, China.

ABSTRACT
Radopholus similis is a migratory plant-parasitic nematode that causes severe damage to many agricultural and horticultural crops. Calreticulin (CRT) is a Ca2+-binding multifunctional protein that plays key roles in the parasitism, immune evasion, reproduction and pathogenesis of many animal parasites and plant nematodes. Therefore, CRT is a promising target for controlling R. similis. In this study, we obtained the full-length sequence of the CRT gene from R. similis (Rs-crt), which is 1,527-bp long and includes a 1,206-bp ORF that encodes 401 amino acids. Rs-CRT and Mi-CRT from Meloidogyne incognita showed the highest similarity and were grouped on the same branch of the phylogenetic tree. Rs-crt is a multi-copy gene that is expressed in the oesophageal glands and gonads of females, the gonads of males, the intestines of juveniles and the eggs of R. similis. The highest Rs-crt expression was detected in females, followed by juveniles, eggs and males. The reproductive capability and pathogenicity of R. similis were significantly reduced after treatment with Rs-crt dsRNA for 36 h. Using plant-mediated RNAi, we confirmed that Rs-crt expression was significantly inhibited in the nematodes, and resistance to R. similis was significantly improved in transgenic tomato plants. Plant-mediated RNAi-induced silencing of Rs-crt could be effectively transmitted to the F2 generation of R. similis; however, the silencing effect of Rs-crt induced by in vitro RNAi was no longer detectable in F1 and F2 nematodes. Thus, Rs-crt is essential for the reproduction and pathogenicity of R. similis.

No MeSH data available.


Related in: MedlinePlus

Neighbor-joining phylogenetic tree of CRT proteins.The phylogram was constructed according to the amino acid sequences of 22 CRT proteins from 20 different species using MEGA 5.0. R. similis CRT is underlined. The accession numbers of the sequences are shown in brackets.
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pone.0129351.g002: Neighbor-joining phylogenetic tree of CRT proteins.The phylogram was constructed according to the amino acid sequences of 22 CRT proteins from 20 different species using MEGA 5.0. R. similis CRT is underlined. The accession numbers of the sequences are shown in brackets.

Mentions: A phylogenetic tree was constructed based on the aa sequences of 22 CRT proteins from 20 different species (Fig 2). Rs-CRT from R. similis, Mi-CRT from M. incognita and Pg-CRT from P. goodeyi were grouped on the same branch, suggesting that they have a closer phylogenetic relationship. In addition, Rs-CRT and the other 21 CRT proteins were divided into 6 groups: animal parasitic nematodes, free-living nematodes, plant parasitic nematodes, insects, mammals, and plants.


A Nematode Calreticulin, Rs-CRT, Is a Key Effector in Reproduction and Pathogenicity of Radopholus similis.

Li Y, Wang K, Xie H, Wang YT, Wang DW, Xu CL, Huang X, Wang DS - PLoS ONE (2015)

Neighbor-joining phylogenetic tree of CRT proteins.The phylogram was constructed according to the amino acid sequences of 22 CRT proteins from 20 different species using MEGA 5.0. R. similis CRT is underlined. The accession numbers of the sequences are shown in brackets.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465493&req=5

pone.0129351.g002: Neighbor-joining phylogenetic tree of CRT proteins.The phylogram was constructed according to the amino acid sequences of 22 CRT proteins from 20 different species using MEGA 5.0. R. similis CRT is underlined. The accession numbers of the sequences are shown in brackets.
Mentions: A phylogenetic tree was constructed based on the aa sequences of 22 CRT proteins from 20 different species (Fig 2). Rs-CRT from R. similis, Mi-CRT from M. incognita and Pg-CRT from P. goodeyi were grouped on the same branch, suggesting that they have a closer phylogenetic relationship. In addition, Rs-CRT and the other 21 CRT proteins were divided into 6 groups: animal parasitic nematodes, free-living nematodes, plant parasitic nematodes, insects, mammals, and plants.

Bottom Line: The reproductive capability and pathogenicity of R. similis were significantly reduced after treatment with Rs-crt dsRNA for 36 h.Using plant-mediated RNAi, we confirmed that Rs-crt expression was significantly inhibited in the nematodes, and resistance to R. similis was significantly improved in transgenic tomato plants.Plant-mediated RNAi-induced silencing of Rs-crt could be effectively transmitted to the F2 generation of R. similis; however, the silencing effect of Rs-crt induced by in vitro RNAi was no longer detectable in F1 and F2 nematodes.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Plant Nematology and Research Center of Nematodes of Plant Quarantine, Department of Plant Pathology, South China Agricultural University, Guangzhou, Guangdong, 510642, China.

ABSTRACT
Radopholus similis is a migratory plant-parasitic nematode that causes severe damage to many agricultural and horticultural crops. Calreticulin (CRT) is a Ca2+-binding multifunctional protein that plays key roles in the parasitism, immune evasion, reproduction and pathogenesis of many animal parasites and plant nematodes. Therefore, CRT is a promising target for controlling R. similis. In this study, we obtained the full-length sequence of the CRT gene from R. similis (Rs-crt), which is 1,527-bp long and includes a 1,206-bp ORF that encodes 401 amino acids. Rs-CRT and Mi-CRT from Meloidogyne incognita showed the highest similarity and were grouped on the same branch of the phylogenetic tree. Rs-crt is a multi-copy gene that is expressed in the oesophageal glands and gonads of females, the gonads of males, the intestines of juveniles and the eggs of R. similis. The highest Rs-crt expression was detected in females, followed by juveniles, eggs and males. The reproductive capability and pathogenicity of R. similis were significantly reduced after treatment with Rs-crt dsRNA for 36 h. Using plant-mediated RNAi, we confirmed that Rs-crt expression was significantly inhibited in the nematodes, and resistance to R. similis was significantly improved in transgenic tomato plants. Plant-mediated RNAi-induced silencing of Rs-crt could be effectively transmitted to the F2 generation of R. similis; however, the silencing effect of Rs-crt induced by in vitro RNAi was no longer detectable in F1 and F2 nematodes. Thus, Rs-crt is essential for the reproduction and pathogenicity of R. similis.

No MeSH data available.


Related in: MedlinePlus