Profiling of differentially expressed genes in hepatopancreas of white shrimp (Litopenaeus vannamei) exposed to long-term low salinity stress
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- 作者：Weihua Gao ; Beiping Tan ; Kangsen Mai ; Shuyan Chi ; Hongyu Liu ; Xiaohui Dong ; Qihui Yang
- 关键词：Litopenaeus vannamei ; Hepatopancreas ; Stress ; Suppression subtractive hybridization ; mRNA expression
- 刊名：Aquaculture
- 出版年：2012
- 出版时间：5 October, 2012
- 年：2012
- 卷：364-365
- 期：Complete
- 页码：186-191
- 全文大小：284 K
文摘
The Pacific white shrimp, Litopenaeus vannamei, is a euryhaline crustacean capable of tolerating a wide range of ambient salinity (0.5-40 psu). To investigate the effect of long-term low salinity stress on gene expression in the hepatopancreas in shrimp, we performed suppression subtractive hybridization (SSH) in juvenile L. vannamei exposed to long-term low salinity. The shrimp (initial body weight, 0.27 ¡À 0.01 g) were cultured at salinity 2 psu and salinity 30 psu for 56 days. We then constructed forward and reverse subtractive cDNA libraries. We used bioinformatics tools and vector screening to select a total of 200 (80 from forward, 120 from reverse) randomly selected clones over 100 nucleotides in length for further analysis. Nineteen contigs and 54 singletons were generated from a total of 73 consensuses. The consensuses, upon a sequence homology search using BLASTX (NCBI), revealed that 24.66 % (18/73) of them had no significant match to reported sequences in the database, suggesting that they had not previously been found and that they were probably associated with stress-regulated functions. The remaining 75.34 % (55/73) of the consensuses encoded proteins were matched to a wide range of functions including immune-related functions, metabolism, ribosomal activity, transfer activity, and apoptosis. The most common group in these SSH libraries was immune-related proteins and enzymes (11/17). Quantitative RT-PCR results confirmed that the relative expression of 5 differentially expressed genes encoding hemocyanin, chitinase, ecdysteroid-regulated protein, trypsin and chymotrypsin 1 was decreased 2-, 1.45-, 11.11-, 1.33- and 1.54-fold, respectively, in the reverse library. This subtractive cDNA library provides a basis for the study of the genetic response of shrimp to environmental stress.