Expression profile of two HSP70 chaperone proteins in response to extreme thermal acclimation in Xestia c-nigrum (Lepidoptera: Noctuidae)

Authors

  • Ling Wang Northeast Agricultural University, College of Agriculture, Heilongjiang Province, Harbin 150030, China
  • Shuai Yang Northeast Agricultural University, College of Agriculture, Heilongjiang Province, Harbin 150030, China Virus-Free Seedling Research Institute, Heilongjiang Academy of Agricultural Sciences, Heilongjiang Province, Harbin, 150086, China
  • Lanlan Han Northeast Agricultural University, College of Agriculture, Heilongjiang Province, Harbin 150030, China
  • Kuijun Zhao Northeast Agricultural University, College of Agriculture, Heilongjiang Province, Harbin 150030, China
  • Lefu Ye Northeast Agricultural University, College of Agriculture, Heilongjiang Province, Harbin 150030, China

Keywords:

heat shock proteins 70 (HSP70s), expression profile, thermal stress, Xestia c-nigrum

Abstract

Heat shock proteins 70 (HSP70s) are highly evolutionarily conserved and play important roles in helping insects survive under extreme temper­atures. In this study, Xestia c-nigrum (L.) (Lepidoptera: Noctuidae) was stressed at various temperatures, and the impacts on thermotolerance and cold endurance were examined. Two complementary deoxyribonucleic acid (cDNA) clones encoding heat shock cognate 70 (HSC70) and inducible heat shock protein 70 (HSP70), named Xc-HSC70 and Xc-HSP70, respectively, were isolated from X. c-nigrum using reverse transcrip­tase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Amino acid sequence analysis indicated that Xc-HSC70 and Xc-HSP70 contained the signature sequences of the HSP70 family, and phylogenetic analysis showed that Xc-HSP70s were consistent with the known insect classification. Genomic DNA structure analysis revealed that the Xc-HSC70 gene contained 8 introns that all conformed to the GT/AG rule, whereas the Xc-HSP70 gene did not contain any introns in the coding region. Fluorescent real-time quantitative RT-PCR re­vealed that both genes were significantly up-regulated under thermal stress and cold stress. The expression patterns indicated that Xc-HSP70 was heat inducible and Xc-HSC70 was constitutively expressed. The temperature response was more intense for Xc-HSP70 than for Xc-HSC70, regardless of the temperature or developmental stage; however, the expression of Xc-HSC70 and Xc-HSP70 was not altered in the pupa. Taken together, the expression profiles of both genes (Xc-HSP70s) provide good support to the insect for coping with adverse conditions.

 

Resumen

Proteínas de choque térmico 70 (Hsp70s) están evolutivamente muy conservadas y juegan un papel importante en ayudar a los insectos a sobrevivir las temperaturas extremas. En este estudio, se puso Xestia c-nigrum (L.) (Lepidoptera: Noctuidae) bajo el estrés de diferentes temperaturas, y se examinó el impacto sobre la termotolerancia y resistencia al frío. Se aislaron dos clones de ácido desoxirribonucleico (ADNc) complementarios que codifican el cognado choque térmico 70 (HSC70) y la proteína de choque térmico inducible 70 (HSP70), llamado Xc-HSC70 y Xc-HSP70, respectivamente, a partir de X. c-nigrum mediante la cadena de polimerasa con transcriptasa inversa reacción (RT-PCR) y la amplificación rápida de extremos de ADNc (RACE). El análisis de la secuencia de aminoá­cidos indicó que Xc-HSC70 y Xc-HSP70 contenían las secuencias de la firma de la familia HSP70, y el análisis filogenético demostró que Xc-Hsp70s es consistente con la clasificación conocida del insecto. El análisis genómico de la estructura del ADN reveló que el gen Xc-HSC70 contenía 8 intrones que todos se ajustaban a la regla GT/AG, mientras que el gen Xc-HSP70 no contenía intrones en la región codificante. El análisis de RT-PCR fluorescente cuantitativa en tiempo real reveló que ambos genes fueron significativamente regulados hacia arriba bajo estrés térmico y estrés por frío. Los patrones de expresión indicaron que Xc-HSP70 fue inducible por el calor y Xc-HSC70 se expresó constitutivamente. La respuesta de la temperatura fue más intensa para Xc-HSP70 que para Xc-HSC70, independientemente de la temperatura o el estadio de desarrollo; sin embargo, la expresión de Xc-HSC70 y Xc-HSP70 no se alteró en la pupa. Tomados en conjunto, los perfiles de expresión de ambos genes (XC-Hsp70s) proveen un buen apoyo para el insecto para hacer frente a las condiciones adversas.

 

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Issue

Vol. 98, No. 2 (June 2015)

Section

Research Papers

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