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Bph32 , a novel gene encoding an unknown SCR domain-containing protein, confers resistance against the brown planthopper in rice

View Article: PubMed Central - PubMed

ABSTRACT

An urgent need exists to identify more brown planthopper (Nilaparvata lugens Stål, BPH) resistance genes, which will allow the development of rice varieties with resistance to BPH to counteract the increased incidence of this pest species. Here, using bioinformatics and DNA sequencing approaches, we identified a novel BPH resistance gene, LOC_Os06g03240 (MSU LOCUS ID), from the rice variety Ptb33 in the interval between the markers RM19291 and RM8072 on the short arm of chromosome 6, where a gene for resistance to BPH was mapped by Jirapong Jairin et al. and renamed as “Bph32”. This gene encodes a unique short consensus repeat (SCR) domain protein. Sequence comparison revealed that the Bph32 gene shares 100% sequence identity with its allele in Oryza latifolia. The transgenic introgression of Bph32 into a susceptible rice variety significantly improved resistance to BPH. Expression analysis revealed that Bph32 was highly expressed in the leaf sheaths, where BPH primarily settles and feeds, at 2 and 24 h after BPH infestation, suggesting that Bph32 may inhibit feeding in BPH. Western blotting revealed the presence of Pph (Ptb33) and Tph (TN1) proteins using a Penta-His antibody, and both proteins were insoluble. This study provides information regarding a valuable gene for rice defence against insect pests.

No MeSH data available.


Western blotting of Bph32 protein expression in E. coli BL21T1R. M1, Precision Plus protein standard; 1, 2, and 3, whole cell lysate, supernatants and sediments of E. coli cells harbouring the control pClod l, respectively; 4, 5, and 6, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0933-2(Ptb33, Pph), respectively; 7, 8, and 9, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0934-1(TN1:Tph), respectively; M2, Perfect Protein Marker.
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f4: Western blotting of Bph32 protein expression in E. coli BL21T1R. M1, Precision Plus protein standard; 1, 2, and 3, whole cell lysate, supernatants and sediments of E. coli cells harbouring the control pClod l, respectively; 4, 5, and 6, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0933-2(Ptb33, Pph), respectively; 7, 8, and 9, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0934-1(TN1:Tph), respectively; M2, Perfect Protein Marker.

Mentions: Western blotting revealed that the Pph (Pyb33) and Tph (TN1) proteins were successfully expressed in Escherichia coli (E. coli) BL21T1R and could be detected using a Penta-His antibody (Fig. 4). The two proteins were insoluble and differed slightly in size.


Bph32 , a novel gene encoding an unknown SCR domain-containing protein, confers resistance against the brown planthopper in rice
Western blotting of Bph32 protein expression in E. coli BL21T1R. M1, Precision Plus protein standard; 1, 2, and 3, whole cell lysate, supernatants and sediments of E. coli cells harbouring the control pClod l, respectively; 4, 5, and 6, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0933-2(Ptb33, Pph), respectively; 7, 8, and 9, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0934-1(TN1:Tph), respectively; M2, Perfect Protein Marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120289&req=5

f4: Western blotting of Bph32 protein expression in E. coli BL21T1R. M1, Precision Plus protein standard; 1, 2, and 3, whole cell lysate, supernatants and sediments of E. coli cells harbouring the control pClod l, respectively; 4, 5, and 6, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0933-2(Ptb33, Pph), respectively; 7, 8, and 9, whole cell lysate, supernatants and sediments of E. coli cells harbouring CDG0934-1(TN1:Tph), respectively; M2, Perfect Protein Marker.
Mentions: Western blotting revealed that the Pph (Pyb33) and Tph (TN1) proteins were successfully expressed in Escherichia coli (E. coli) BL21T1R and could be detected using a Penta-His antibody (Fig. 4). The two proteins were insoluble and differed slightly in size.

View Article: PubMed Central - PubMed

ABSTRACT

An urgent need exists to identify more brown planthopper (Nilaparvata lugens Stål, BPH) resistance genes, which will allow the development of rice varieties with resistance to BPH to counteract the increased incidence of this pest species. Here, using bioinformatics and DNA sequencing approaches, we identified a novel BPH resistance gene, LOC_Os06g03240 (MSU LOCUS ID), from the rice variety Ptb33 in the interval between the markers RM19291 and RM8072 on the short arm of chromosome 6, where a gene for resistance to BPH was mapped by Jirapong Jairin et al. and renamed as “Bph32”. This gene encodes a unique short consensus repeat (SCR) domain protein. Sequence comparison revealed that the Bph32 gene shares 100% sequence identity with its allele in Oryza latifolia. The transgenic introgression of Bph32 into a susceptible rice variety significantly improved resistance to BPH. Expression analysis revealed that Bph32 was highly expressed in the leaf sheaths, where BPH primarily settles and feeds, at 2 and 24 h after BPH infestation, suggesting that Bph32 may inhibit feeding in BPH. Western blotting revealed the presence of Pph (Ptb33) and Tph (TN1) proteins using a Penta-His antibody, and both proteins were insoluble. This study provides information regarding a valuable gene for rice defence against insect pests.

No MeSH data available.