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| The Effect of Imidazole Derivative KK-42 on Expression of Chitinase Gene in Oriental River Prawn Macrobrachium nip ponense |
| WANG Pei,L U· · Yan-jie,L·U· Li,YANG Hong,NING Qian-ji |
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Abstract This study was designed to determine the possible molecular mechanism of imidazole derivative KK‐42 acting for increasing survival rate of oriental river prawn Macrobrachium nipponense infected with Aeromonas hydrophila .The part sequence of oriental river prawn chitinase‐3 was firstly cloned and the spatio‐temporal expression of the gene as well as its activity were measured .The prawn with body length 3 .5—5 .0 cm was soaked for 1 min in KK‐42 solution at a concentration of 1 .95 × 10‐4 mol/L (KK‐42 treat‐ment group 1) or 0 (control group 1) ,separatively .The prawn in KK‐42 treatment group 1 was injected individually with A .hydrophila suspension (KK‐42 treatment group‐A .hydrophila) or saline (KK‐42 treatment group 2) into the ventral sinus at 12 h after KK‐42 treatment and the prawn in control group 1 was divided into control group‐A .hydrophila and control group 2 ,according to the same way .Then the survival rate was surveyed at different time ,and the prawn chitinase‐3 mRNA level and activity derived from different tissues were measured .Results showed that the survival rate was dramatically increased by 133% in KK‐42 treatment group‐A .hydrophila than that in the control group‐A .hydrophila .Sequence comparison indicated that the prawn chitinase‐3 deduced amino acid sequence of M .nipponense had an o‐verall similarity of 90% to that of other crustaceans .Real‐time PCR analysis revealed that the praw n chiti‐nase‐3 was mainly expressed in hepatopancreas ,and higher at premolt .Compared with the control group 1 ,the changes in the prawm chitinase‐3 mRNA level and activity from KK‐42 treatment group was found to be an up‐regulation about 150% and 75% ,respectively .In KK‐42 treatment‐A .hydrophila group ,the prawn chitinase‐3 mRNA transcripts were significantly higher at 12—48 h ,the maximum at 24 h with an increase by 304% ,than that in control group‐A .hydrophila and its activity increased by 48% at 24 h .In conclusion ,imidazole derivative KK‐42 pretreatment can significantly up‐regulate the prawn chitinase‐3 ex‐pression as well as its activity in hepatopancreas of prawn infected with A .hydrophila ,which is likely one of the molecular mechanisms of imidazole derivative KK‐42 acting for increasing survival rate of the prawn infected with A .hydrophila .
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Received: 25 December 2014
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