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| Comparison of the Effects of Different Enhancers of the Mbk-2 Gene on Embryonic Development in Cryptobacterium Hidradenum |
| ZHANG Jundi, et al |
| Langfang People's Hospital, Hebei Langfang 065000, China |
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Abstract Objective: To Explore the effects of mbk-2 and its enhancers on nematode embryonic development using the mbk-2 mutant Cryptobacterium hidradenum as a model organism. Methods: Targeted silencing of mbk-2 enhancers pptr-1, pptr-2, rsa-1 and rsa-2 by RNAi technology were used to observe changes in nematode egg unhatched rate and embryonic developmental stages after silencing of mbk-2 enhancers. Results: The unhatched rate of mbk-2 mutant nematode eggs was 1.56 ± 0.25%, significantly higher than that of the N2 wild-type group (0) (P<0.05); the unhatched rates of N2 wild-type nematode eggs fed with pptr-2, rsa-1 and rsa-2 RNAi bacteria were 5.77 ± 0.24%, 4.40 ± 0.03% and 6.03 ± 1.19%, respectively, all significantly higher (P<0.05) than the normal group of N2 fed with L4440 bacteria (0). The unhatched rates of mbk-2 mutant nematode eggs fed by pptr-1, pptr-2, rsa-1 and rsa-2 RNAi bacteria were 3.78±0.14%, 75.53±2.29%, 41.79±0.76% and 51.65±8.23%, respectively, all of which were significantly higher than the unhatched rates of mbk-2 mutant nematode eggs (P<0.05), with the pptr-2 (75.53±2.29%) RNAi group having the highest unhatched rate. The unhatched rates of mbk-2 mutant nematode eggs fed by pptr-1, pptr-2, rsa-1 and rsa-2 RNAi bacteria were 3.78±0.14%, 75.53±2.29%, 41.79±0.76% and 51.65±8.23%, respectively, all of which were significantly higher than the unhatched rates of mbk-2 mutant nematode eggs (P<0.05). with the pptr-2 (75.53±2.29%) RNAi group having the highest unhatched rate. DIC images showed that the rsa-2 RNAi group was significantly abnormal compared to the mbk-2 mutant nematode at the unicellular stage of embryonic development (spindle establishment, absence of prokaryotic encounters close to the posterior of the cell and absence of prokaryotic transfer to the centre of the embryo) (P<0.05). The pptr-2 RNAi group was significantly abnormal compared to the mbk-2 mutant nematode at the four-cell stage of embryonic development (large differences in cell size, multiple nuclei and abnormal intercellular contacts) (P<0.05). Conclusion: The mbk-2 and its enhancer play an important regulatory role in nematode embryonic development.
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2022, Vol. 28 
