重要水产养殖虾蟹类的多倍体诱导及性别控制研究

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	重要水产养殖虾蟹类的多倍体诱导及性别控制研究
其他题名	Polyploid induction and sex control in several economically important crustacean species
	张成松
学位类型	博士
	2009-05-25
学位授予单位	中国科学院海洋研究所
学位授予地点	海洋研究所
关键词	中国明对虾凡纳滨对虾中华绒螯蟹三倍体雌性化
摘要	本论文选取中国明对虾、凡纳滨对虾和中华绒螯蟹这三个我国重要的养殖品种进行了多倍体和雌性化控制的研究，优化了中国明对虾和中华绒螯蟹的多倍体诱导方法及凡纳滨对虾雌性化控制的关键参数，并在此基础上研究了三倍体中国明对虾在早期生长阶段的生产性状和生物能量学特征。利用热休克的方法抑制第一极体释放可以获得高比例的三倍体；热休克显著降低了中国明对虾的变态存活率，而倍性本身对其影响不明显；根据部分形态学指标发展了中国明对虾倍性判别公式，其判别准确率在90%以上。倍性操作对中国明对虾幼虾阶段的急性盐度突变的适应能力及摄食率无明显影响，但其对三倍体中国明对虾的养殖成活率、生长率、饵料转化效率及蜕皮周期有显著影响（P<0.05）；二倍体对虾的最适生长盐度为20‰，而三倍体对虾为30‰。倍性操作对中国明对虾的基础代谢水平和能量收支模式有显著的影响。三倍体对虾的耗氧率及排氨率显著高于二倍体对照（P<0.05）；但在性腺发育以前，三倍体对虾分配于生长的能量份额明显低于二倍体（P<0.05）；而在性腺发育早期其分配于生长的能量份额与二倍体对虾无明显差异（P>0.05）。初步确定了一个不受温度、遗传背景等影响的可用于大批量生产河蟹三倍体的起始处理时刻的形态学标记-受精膜举起，大大提高了中华绒螯蟹三倍体诱导效率。凡纳滨对虾受精卵对温度的耐受力随发育而提高；在原肠中后期，热休克处理（39℃处理3min）可以显著提高诱导群体的雌性比例。
其他摘要	Crustacean cultivation (especially to shrimp and crab), which has contributed largely to the employment of rural population and the augment of rural incomes, is one of the economically important industries in China, it has brought us huge social and economic benefit. Unfortunately, for the reasons of serious diseases, idioplasmic degeneration and environmental deterioration, the development of culture in shrimps and crabs encountered prodigious handicap in China in recently years. So the genetic improvement to the crustacean species is imperative. Previous researches on polyploid breeding and mono-sex culture of aquatic animal have been proved to be successful in increasing yield and improving quality. In the present thesis, the Chinese shrimp (Fenneropenaeus chinensis), the Pacific white shrimp (Litopenaeus vannamei) and the Chinese mitten crab (Eriocheir japonica sinensis) were selected as the experimental animals. Heat shock or potassium chloride (KCl) was employed to induce polyploidy in F. chinensis or E. japonica sinensis, and the feminization control was carried out in L. vannamei by heat shock. Heat shock was employed to inhibit the releasing of the first polar body to produce meiosis Ⅰtriploid F. chinensis. At 15℃, heat shock (temperature, 29-31℃; duration, 8-10min ) successfully induced MⅠtriploids when treated at 7min post spawning, and the highest triploid rate is about 100 percent. No aneuploid and tetraploid were observed in treated samples by inhibition of the 1st polar body. Ploidy itself did not affect shrimp larvae survival during metamorphosis, but heat shock significantly decreased the survival rate. Ploidy manipulation significantly affected the morphological characteristics of F. chinensis, the body shape of triploid shrimp is dumpy, but the diploid shrimp is slender. Based on morphological parameters, discriminating formulae for triploid and diploid shrimp at grow-out stage were developed and could be used to distinguish triploids from diploids. Triploid Chinese shrimp Fenneropenaeus chinensis were reared up by heat shock inducement to inhibit meiosisⅡ. The growth performances in full-sib diploid and triploid F. chinensis were compared to evaluate the effect of ploidy manipulation at juvenile stage under laboratory conditions. It showed that there was no significant difference (P>0.05) in tolerance observed in triploid and diploid shrimp due to abrupt salinity changes. The lethal salinity for 50% of the individuals in 96h at 23-25℃ was about 2‰ in both triploids and diploids. Ploidy significantly influenced (P<0.05) the survival rate, special growth rate, feed conversion efficiency and intermolt period of F. chinensis, but there were no difference of the feeding rate between the two ploidy levels. Based on the survival and growth data, the optimum salinity for the culture of diploid F. chinensis should be 20‰ and for triploids it should be 30‰. At the early stage during culture, triploid F. chinensis juvenile showed no growth superiority on their diploid controls . The effects of ploidy manipulation on bioenergetics of F. chinensis were studied under laboratory conditions, and the bioenergetics’ bases to explain the different growth performance during culture period were proved up. At immature stage, triploid shrimp exhibited higher oxygen consumption rate than the diploids at the range of acclimation salinities (15-35ppt) and temperatures (26-32C), and higher ammonia-N excretion rate at some salinity level (25 and 35ppt) and temperature level (32C). Triploid shrimp have lower energy allot for growth than that of the diploids (P<0.05). The tendencies of environmental factors on standard metabolic rate were similar between the two ploidy levels. At the early stage of gonad development, there was no difference of energy budget between triploid and diploid shrimp (P>0.05), but the triploids exhibited higher metabolic rate (P<0.05). This is the first report of bioenergetics in triploid shrimp and it likely explain the prior experimental results which with difference of growth performances between triploid and diploid shrimp. Based on morphological observation of deciduous and normal attached fertilized eggs of E. japonica sinensis, a potential morphological marker (the apparition of the plasma membrane and apparition of maximum distance between the plasma membrane and yolk mass) which can be used as the optimum starting time shocked by KCl was determined. This is the first report of morphological dynamics of early development of fertilized eggs from Chinese mitten crab, and later repeated experiments indicated that the morphological marker can be used as an indicator for highly efficient triploid induction with KCl in this crab species. To mono-sex culture, heat shock was employed to feminize the L. vannamei at different stages of embryonic development. It showed that the thermotolerance of shrimp embryos raised as the development advanced, at middle and later stages of gastrulae, heat shock (temperature, 39℃; duration, 3min) significantly increased the female ratio in the treated populations. This is the first report of feminization in L. vannamei treated with heat shock.
页数	119
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/1247
专题	海洋环流与波动重点实验室实验海洋生物学重点实验室
推荐引用方式 GB/T 7714	张成松. 重要水产养殖虾蟹类的多倍体诱导及性别控制研究[D]. 海洋研究所. 中国科学院海洋研究所,2009.

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