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Triploid Induction in Chlamys Farreri by Hypotonic Treatment and the Comparision with Other Treatment Methods

Author: ZhangChenChen
Tutor: WangZhaoPing
School: Ocean University of China
Course: Aquaculture
Keywords: C. farreri Triploid Hypotonic induction Fertilized egg Ultrastructure
CLC: S917.4
Type: Master's thesis
Year: 2010
Downloads: 46
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Abstract


In this paper, Chlamys farreri (Chlamys farreri) study by hypotonic treatment to inhibit the release of the second polar body (PB2) C. farreri fertilized egg to produce triploid. Explore hypotonic treatment induced conditions, triploid induction and training, as well as comparison with other methods, and by electron microscopy ultrastructural changes of the fertilized egg after hypotonic processing, more times Sports species theory provides new experimental and practical basis. The study is divided into the following sections: 1, hypotonic method of hypotonic induced the C. farreri triploid optimal induction parameters, including through the different induction parameters: water temperature 20 ℃, respectively, for different salinity treatment (8 -20), start processing time (fertilization after 15-40min) and different the ongoing processing time (10-25min) experiment. The results show that: hypotonic salinity S = 14 after fertilization 30min begin processing, continuous processing 20min induction rate of up to (92.36 ± 2.41)%. Chlamys farreri triploid cultivation hypotonic method Chlamys farreri induced triploid optimal induction conditions, the D-type larval stage, treated larvae showed significant growth advantage (P lt; 0.01), not only fast growth and development to metamorphosis, the individual is larger than diploid. The newly hatched triploid D-shaped larvae induction rate was 90.15%, the larvae of the subsequent induction rate decreased gradually decreased to 60.92% when the metamorphosis attached. 3, the hypotonic induction Chlamys farreri triploid Compare ① hypotonic with other methods: using this study to determine the best program; ② 6-DMAP: concentration of 60mg / L, 40 ~ 50% of fertilized eggs first polar body processing time 15min; ③ heat shock: 30 ℃, 40 to 50% of the fertilized egg first polar body, processing time 15min; ④ cold shock: 1 ℃ ,40-50% fertilized egg first polar body processing time 20min. The four methods were compared, triploid induction rate hypotonic treatment group is the highest (91.43 ± 4.14)% ,6-DMAP treatment followed by group (91.25 ± 3.49)%, lower heat shock and cold shock induction rate were (25.85 ± 4.94)% and (21.86 ± 2.04)%; hypotonic induction effect of 6-DMAP was no significant difference, P gt; 0.05; induced by hypotonic with 6-DMAP treated better than heat shock and cold shock group, the difference was significant, P lt; 0.01 4 the different hypotonic configuration induced the C. farreri triploid comparison water temperature 20 ° C, oxytocin to obtain the same batch of sperm and ovum, fertilized at the same time, using KCl and distilled water the treatment liquid, of the paired salinity 14 NaCl with distilled water dubbed salinity 14 treatment liquid and salinity 14 is paired with a salinity of 31 natural sea water and distilled water treatment solution was subjected to hypotonic treatment, each group could varying degrees induction of the C. farreri fertilized eggs produce triploid triploid induction rate, respectively: (90.01 ± 3.27)%, (90.07 ± 2.64)%, (91.43 ± 4.14)% hypotonic induced effects in each group were no significant differences , P gt; 0.05. 5, The electron microscope the hypotonic treatment Chlamys farreri fertilized eggs using scanning electron microscopy ultrastructure observation the C. farreri fertilized egg hypotonic treatment, contrast changes in the structure of the fertilized egg before and after the treatment, the results show that the hypotonic treatment can cause fertilized egg changes: the induction group fertilized eggs significant expansion, the volume is greater than the control group, and the irregular shape; When the process ends after soaking into the normal seawater, fertilized eggs with a certain degree of recovery: the shape of the treatment groups tends to rule, the volume of the control group had no significant difference. Ultrastructure observation by electron microscopy of the hypotonic treatment Chlamys farreri fertilized egg, comparing before and after treatment zygote changes in the structure, the results show that the the hypotonic treatment can cause a fertilized egg changes: destruction of the egg membrane, plasma membrane occurs wrinkle compression and partial dissolution of the extracellular matrix thinning, the yolk granules shrinkage deformation; fertilized eggs have a certain degree of recovery after the end of the treatment into normal seawater immersion: the plasma membrane becomes smooth, extracellular matrix filling yolk particle shape comparison rules .

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
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