Ecological Safety Assessment Of Transgenic Cry1Ab/Ac Insect-resistant Rice And Its Toxic Protein On Misgurnus Anguillicaudatus

In this study, three kinds of ways, which are field feeding, pollen feeding,poisoned rice straw powder feeding, were employed to assess the effect of transgenicCry1Ab/Ac insect-resistant rice (HH1) on the safety of the loach (Misgurnusanguillicaudatus), and the non-transgenic rice (MH63) was set as a control. TheCry1Ab/Ac protein content, growth and development, physiological enzyme, bloodcell damage and digestive histology of the loach were determined as the indexes. Themain results are as follows:1. Content of toxic protein in transgenic Cry1Ab/Ac rice and various organs ofMisgurnus anguillicaudatusChoosing Cry1Ab/Cry1Ac kits from EnvirLogix, we applied enzyme-linkedimmunosorbent assay (ELISA) to test the rice at different growth stages. ElISAresults showed that the expression of Cry1Ab/Ac protein in different organs of HH1rice was well. The content of Cry1Ab/Ac protein in mature stage was higher thanheading stage, and heading stage higher than jointing stage.At the stages of jointing, heading and mature, content of Cry1Ab/Ac protein ingill, intestine, stomach, muscle and gonad of loach was lower than the minimumdetectable value, which is0.2ng/g.2. Effects of transgenic Cry1Ab/Ac rice on the growth of MisgurnusanguillicaudatusBy weighed the fresh weight and counted parameters, we can see from thestatistics results that weight gain rate, condition factor, viscera somatic index,hepatosomatic index and mature coefficient of loach in HH1rice field environmenthad no negative effect in2012. The HH1rice field environment in2013had noobvious effect on weight rate, condition factor, hepatosomatic index, viscera somaticindex and mature coefficient of loach at the stage of jointing, heading and mature（P>0.05）.1%,5%,10%,20%and40%of HH1pollen were respectively added to fodder,which had no negative effect on weight gain rate, condition factor, viscera somaticindex, hepatosomatic index, kidneymatic index, mature coefficient of loach,compared with the control（P>0.05）.10,50,100and200mg/L HH1straw powder in water had no effect on weightgain rate, condition factor, viscera somatic index, hepatosomatic index and maturecoefficient of loach（P>0.05）.3. Effects of transgenic Cry1Ab/Ac rice on activity of biological enzyme ofMisgurnus anguillicaudatus Pyrogallol autoxidation, UV spectrophotometry, CDNB, Folin phenol methodand Ellman method were separately used to determine the activity of SOD、CAT、GST、Proteolytic and AChE of loach. Compared with control, the activity of SOD,CAT and GST in liver of loach showed no significant difference in2012（P>0.05）.The different growth stages of HH1in2013had no effect on SOD, CAT, GST ofliver, kidney and gill of loach. HH1rice field had no difference on stomach, foregut,midgut and hindgut poteolytic enzyme of loach, compared with the control. Theactivity of AChEbrain, liver and muscle of loach were not affect by the HH1rice（P>0.05）.After loach fed with0~40%HH1pollen, activity of SOD, CAT and GST in itsliver and kidney had no obvious change. HH1pollen had no effect on stomach andintestinal poteolytic of loach. Activity of AChE of brain, liver and muscle were notaffected by HH1pollen（P>0.05）.The0~200mg/L HH1rice straw powder had no obvious effect on the antioxidantenzyme (SOD, CAT, GST), poteolytic enzyme and AChE of loach, compared withcontrol （P>0.05）.4. Effects of transgenic Cry1Ab/Ac rice on blood routine and red cell ofMisgurnus anguillicaudatusWe chose Nanjing Jiancheng methemoglobin kit and blood smear method tostudy the blood of loach. The results showed that HH1paddy environment had nosignificant negative on the content of Hb and MHb, as well as blood cell nucleus ofloach in2012and2013（P>0.05）.HH1pollen had no obvious effect on Hb and MHb of loach. The micronucleusrate, nuclear abnormal rate, nuclear convex rate and nuclear unequal division rate of10~40%HH1pollen groups showed no significant difference with control（P>0.05）.10~200mg/L HH1straw powder did not obviously affect the Hb and MHb, andthe blood red cell nuclear of loach of200mg/L HH1group were not changedsignificantly compared with control（P>0.05）.5. Effects of transgenic Cry1Ab/Ac rice on organization structure of gonad ofMisgurnus anguillicaudatusParaffin section method was used to observe the structure of ovary andhistological of loach. The result showed that HH1rice field environment had nosignificant effect on the gonad of loach（P>0.05）.