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Proteomic analysis of Apis cerana and Apis mellifera larvae fed with heterospecific royal jelly and by CSBV challenge.

Zhang Y, Zhang G, Huang X, Han R - PLoS ONE (2014)

Bottom Line: Heterospecific royal jelly (RJ) breeding in two honeybee species may result in morphological and genetic modification.In Ac larvae, 6 differential expression proteins were identified from heterospecific RJ breeding only, 21 differential expression proteins from CSBV challenge only and 7 differential expression proteins from heterospecific RJ breeding plus CSBV challenge.In Am larvae, 17 differential expression proteins were identified from heterospecific RJ breeding only, 26 differential expression proteins from CSBV challenge only and 24 differential expression proteins from heterospecific RJ breeding plus CSBV challenge.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Entomological Institute, Guangzhou, China.

ABSTRACT
Chinese honeybee Apis cerana (Ac) is one of the major Asian honeybee species for local apiculture. However, Ac is frequently damaged by Chinese sacbrood virus (CSBV), whereas Apis mellifera (Am) is usually resistant to it. Heterospecific royal jelly (RJ) breeding in two honeybee species may result in morphological and genetic modification. Nevertheless, knowledge on the resistant mechanism of Am to this deadly disease is still unknown. In the present study, heterospecific RJ breeding was conducted to determine the effects of food change on the larval mortality after CSBV infection at early larval stage. 2-DE and MALDI-TOF/TOF MS proteomic technology was employed to unravel the molecular event of the bees under heterospecific RJ breeding and CSBV challenge. The change of Ac larval food from RJC to RJM could enhance the bee resistance to CSBV. The mortality rate of Ac larvae after CSBV infection was much higher when the larvae were fed with RJC compared with the larvae fed with RJM. There were 101 proteins with altered expressions after heterospecific RJ breeding and viral infection. In Ac larvae, 6 differential expression proteins were identified from heterospecific RJ breeding only, 21 differential expression proteins from CSBV challenge only and 7 differential expression proteins from heterospecific RJ breeding plus CSBV challenge. In Am larvae, 17 differential expression proteins were identified from heterospecific RJ breeding only, 26 differential expression proteins from CSBV challenge only and 24 differential expression proteins from heterospecific RJ breeding plus CSBV challenge. The RJM may protect Ac larvae from CSBV infection, probably by activating the genes in energy metabolism pathways, antioxidation and ubiquitin-proteasome system. The present results, for the first time, comprehensively descript the molecular events of the viral infection of Ac and Am after heterospecific RJ breeding and are potentially useful for establishing CSBV resistant populations of Ac for apiculture.

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Western blot Analysis of differentially expressed proteins.Six proteins were selected. Proteins were separated on a 12% SDS-PAGE gel and immunoblotted with anti-GAPDH, anti-PKG2, anti-Tpi, anti-FAH, anti-SOD, and anti-FBA. β-actin was used as an internal control. (A) The Western blot images of GAPDH, PKG2, TPI, FAH, SOD, FBA and Actin. The protein spots from Figure 2. (B) Quantification of target protein expression.The relative fold change of GAPDH, PKG2, Tpi, FAH, SOD, FBA (normalized by Actin) was visulized by using Quantity One Software. The expression of GAPDH, SOD, FBA and Tpi were up-regulatedd. On the other hand, FAH and PGK were down-regulated. Error bar is standard deviation.
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pone-0102663-g008: Western blot Analysis of differentially expressed proteins.Six proteins were selected. Proteins were separated on a 12% SDS-PAGE gel and immunoblotted with anti-GAPDH, anti-PKG2, anti-Tpi, anti-FAH, anti-SOD, and anti-FBA. β-actin was used as an internal control. (A) The Western blot images of GAPDH, PKG2, TPI, FAH, SOD, FBA and Actin. The protein spots from Figure 2. (B) Quantification of target protein expression.The relative fold change of GAPDH, PKG2, Tpi, FAH, SOD, FBA (normalized by Actin) was visulized by using Quantity One Software. The expression of GAPDH, SOD, FBA and Tpi were up-regulatedd. On the other hand, FAH and PGK were down-regulated. Error bar is standard deviation.

Mentions: Western blot analysis was conducted to verify the expression of the proteins which are closely related to energy metabolism of heterospecific royal jelly and CSBV resistance, such as GAPDH, PGK, Tpi, FAH, SOD, and FBA. The results are consistent well with the proteomic data (Figure 8).


Proteomic analysis of Apis cerana and Apis mellifera larvae fed with heterospecific royal jelly and by CSBV challenge.

Zhang Y, Zhang G, Huang X, Han R - PLoS ONE (2014)

Western blot Analysis of differentially expressed proteins.Six proteins were selected. Proteins were separated on a 12% SDS-PAGE gel and immunoblotted with anti-GAPDH, anti-PKG2, anti-Tpi, anti-FAH, anti-SOD, and anti-FBA. β-actin was used as an internal control. (A) The Western blot images of GAPDH, PKG2, TPI, FAH, SOD, FBA and Actin. The protein spots from Figure 2. (B) Quantification of target protein expression.The relative fold change of GAPDH, PKG2, Tpi, FAH, SOD, FBA (normalized by Actin) was visulized by using Quantity One Software. The expression of GAPDH, SOD, FBA and Tpi were up-regulatedd. On the other hand, FAH and PGK were down-regulated. Error bar is standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125304&req=5

pone-0102663-g008: Western blot Analysis of differentially expressed proteins.Six proteins were selected. Proteins were separated on a 12% SDS-PAGE gel and immunoblotted with anti-GAPDH, anti-PKG2, anti-Tpi, anti-FAH, anti-SOD, and anti-FBA. β-actin was used as an internal control. (A) The Western blot images of GAPDH, PKG2, TPI, FAH, SOD, FBA and Actin. The protein spots from Figure 2. (B) Quantification of target protein expression.The relative fold change of GAPDH, PKG2, Tpi, FAH, SOD, FBA (normalized by Actin) was visulized by using Quantity One Software. The expression of GAPDH, SOD, FBA and Tpi were up-regulatedd. On the other hand, FAH and PGK were down-regulated. Error bar is standard deviation.
Mentions: Western blot analysis was conducted to verify the expression of the proteins which are closely related to energy metabolism of heterospecific royal jelly and CSBV resistance, such as GAPDH, PGK, Tpi, FAH, SOD, and FBA. The results are consistent well with the proteomic data (Figure 8).

Bottom Line: Heterospecific royal jelly (RJ) breeding in two honeybee species may result in morphological and genetic modification.In Ac larvae, 6 differential expression proteins were identified from heterospecific RJ breeding only, 21 differential expression proteins from CSBV challenge only and 7 differential expression proteins from heterospecific RJ breeding plus CSBV challenge.In Am larvae, 17 differential expression proteins were identified from heterospecific RJ breeding only, 26 differential expression proteins from CSBV challenge only and 24 differential expression proteins from heterospecific RJ breeding plus CSBV challenge.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Entomological Institute, Guangzhou, China.

ABSTRACT
Chinese honeybee Apis cerana (Ac) is one of the major Asian honeybee species for local apiculture. However, Ac is frequently damaged by Chinese sacbrood virus (CSBV), whereas Apis mellifera (Am) is usually resistant to it. Heterospecific royal jelly (RJ) breeding in two honeybee species may result in morphological and genetic modification. Nevertheless, knowledge on the resistant mechanism of Am to this deadly disease is still unknown. In the present study, heterospecific RJ breeding was conducted to determine the effects of food change on the larval mortality after CSBV infection at early larval stage. 2-DE and MALDI-TOF/TOF MS proteomic technology was employed to unravel the molecular event of the bees under heterospecific RJ breeding and CSBV challenge. The change of Ac larval food from RJC to RJM could enhance the bee resistance to CSBV. The mortality rate of Ac larvae after CSBV infection was much higher when the larvae were fed with RJC compared with the larvae fed with RJM. There were 101 proteins with altered expressions after heterospecific RJ breeding and viral infection. In Ac larvae, 6 differential expression proteins were identified from heterospecific RJ breeding only, 21 differential expression proteins from CSBV challenge only and 7 differential expression proteins from heterospecific RJ breeding plus CSBV challenge. In Am larvae, 17 differential expression proteins were identified from heterospecific RJ breeding only, 26 differential expression proteins from CSBV challenge only and 24 differential expression proteins from heterospecific RJ breeding plus CSBV challenge. The RJM may protect Ac larvae from CSBV infection, probably by activating the genes in energy metabolism pathways, antioxidation and ubiquitin-proteasome system. The present results, for the first time, comprehensively descript the molecular events of the viral infection of Ac and Am after heterospecific RJ breeding and are potentially useful for establishing CSBV resistant populations of Ac for apiculture.

Show MeSH
Related in: MedlinePlus