Eur. J. Entomol. 112 (4): 676-687, 2015 | 10.14411/eje.2015.094

Analysis of transcripts of heat shock protein genes in silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Nalavadi CHANDRAKANTH2, Kangayam M. PONNUVEL1,*, Shunmugam M. MOORTHY2, Sirigineedi SASIBHUSHAN1, Vankadara SIVAPRASAD2
1 Genomics Laboratory, Seribiotech Research Laboratory, Carmelaram Post, Kodathi, Bangalore 560 035, India; e-mails: kmpvel@yahoo.com; sasibhushan.s@gmail.com
2 Molecular Biology Laboratory, Central Sericultural Research and Training Institute, Srirampura, Mysore 570008, India; e-mails: chandra.nalavadi@gmail.com; moorthysm68@gmail.com; siva.nsso@gmail.com

Silkworm is a poikilothermic insect, whose growth and development is significantly affected by high temperatures. The current study aimed to identify bivoltine breeds tolerant of the high temperature conditions that occur in the tropics. Percentage pupation at high temperatures and heat shock responses of silkworms were used as measures of thermotolerance. Thermotolerance of 20 silkworm breeds was assessed by rearing them at 36C. Based on percentage pupation, three breeds, namely Nistari (multivoltine), SK4C (bivoltine) and CSR2 (bivoltine) were designated tolerant, moderately tolerant and susceptible, respectively. To understand the heat shock responses and the molecular mechanisms underlying thermotolerance, the tissue specific expression profiles of the nine heat shock protein (Hsp) genes were determined in the three breeds after a heat shock of 1 h at 36C and a 2 h recovery period by performing real-time qPCR. The level of expression of Hsp genes was significantly increased in heat shocked tissues and gradually decreased during the recovery period. The greatest increase in the expression of Hsp genes was recorded in fat body followed by mid gut and silk gland. Of the three breeds, Nistari showed the highest expression of Hsp genes and SK4C a moderate expression relative to CSR2. The qPCR results showed that the transcript levels of sHsp20.4 and 20.1, and Hsp70 were increased by 10.3, 9.7 and 2.3 times, respectively, in Nistari compared to CSR2. Similarly the expression of sHsp20.4 and 20.1, and Hsp70 were increased by 3.5, 2.3 and 1.5 times, respectively in SK4C compared to CSR2. The expression levels of Hsps during heat shock corresponded to the percentage pupation recorded for the three breeds at a high temperature. It is suggested that the Hsps and their levels of expression may play an important role in increasing the survival of silkworm larvae at high temperatures. This study identified SK4C as a bivoltine breed, which is highly tolerant of high temperature measured in terms of percentage pupation (of the bivoltine breeds) and higher levels of expression of Hsp genes compared to CSR2. The importance of SK4C as a thermotolerant bivoltine parent for breeding new bivoltine hybrids tolerant of high temperatures is discussed.

Keywords: Lepidoptera, Bombycidae, Bombyx mori, heat shock proteins, real time qPCR, thermotolerance

Received: December 10, 2014; Accepted: July 27, 2015; Prepublished online: September 22, 2015; Published: November 21, 2015

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