中国对虾育种方法研究与遗传连锁图谱的构建

IOCAS-IR > 海洋环流与波动重点实验室

	中国对虾育种方法研究与遗传连锁图谱的构建
其他题名	Breeding method and genetic linkage maps construction in Fenneropenaeus chinensis
	田燚
学位类型	博士
	2007-05-31
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	中国对虾杂交生长性状存活率回归方程遗传力遗传相关遗传连锁图谱 Aflp
摘要	本研究以中国对虾为材料，以杂交育种和选择育种为目标，进行了系统的中国对虾杂交育种试验、生长性状遗传参数试验及其分子遗传连锁图谱的构建工作。结果表明以不同地理群体杂交作为基础群体，然后采用系统的选择育种方法可以获得较好的选择效果。构建的遗传连锁图谱为中国对虾分子辅助育种提供一定的基础。这些试验结果为中国对虾合理系统的育种工作提供了理论基础和数据支持。其具体结果如下： 1. 试验对中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）的2个群体及其杂交后代不同月龄生长情况和存活率进行了研究，测量体长（TL）、头胸甲长（CL）、头胸甲宽（CW）、第2、3腹节高（HST）、第2、3腹节宽（WST）、体重（BW）和存活率共7个性状，计算各项指标的杂种优势率，并对各性状进行了方差分析和多重比较。其3月龄生长情况和存活率研究结果表明，存活率在乳山湾群体（WYP♀）× 朝鲜半岛南海群体（WKN♂）杂交后代出现杂种劣势外，其他指标都表现出不同程度的杂种优势(4.37%～23.96% )。除了存活率外，杂交后代生长性状均显著高于亲本，乳山湾群体（WYP♀）×朝鲜半岛南海群体（WKN♂）杂交后代高于朝鲜半岛南海群体（WKN♀）× 乳山湾群体（WYP♂）杂交后代，黄渤海水域乳山湾群体高于朝鲜半岛南海群体后代。为确定测量性状与中国对虾体重的相关程度，建立了用体长(X1)，头胸甲宽(X2)，第2、3腹节宽(X3)，头胸甲长(X4)，第2、3腹节高(X5)估计体重的多元回归方程：Y = －2.056 + 0.03X1 + 0.076X2 + 0.078X3 + 0.033X4 + 0.043X5。 2. 中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）2个群体及其杂交后代在4月龄时期的6个生长指标和存活率的杂种优势范围在0.514%到14.95%之间，WYP♀×WKN♂杂交后代在这7个指标中都高于WKN♀×WYP♂杂交后代。5月龄杂交后代也表现出一定程度的杂种优势，其范围在-9.000%～19.090%之间，但头胸甲长、第2、3腹节处高和存活率3个指标出现杂种劣势。不同杂交组合各个阶段生长发育情况和存活率在杂种优势表现出一定的规律。随着月龄的增加，WKN♀×WYP♂杂交后代杂种优势率有所增加，而WYP♀×WKN♂杂交后代的却有所降低。ANOVA分析结果表明，杂交后代在存活率方面与双亲差异不显著。4月龄的分析结果发现杂交后代在WST和BW这2个指标上存在显著差异。LSD多重比较结果显示，WYP♀×WKN♂杂交后代在BW指标上与亲本存在显著差异，在WST指标方面与其他3个组合的后代差异显著。5月龄的数据分析结果发现，杂交后代除体重存在显著差异外，其他各项指标差异均不显著。LSD多重比较结果发现，WKN♀×WYP♂杂交后代体重与其亲本WKN存在显著差异。 3. 对2个野生群体——朝鲜半岛南海岸群体（WKN）和黄渤海群体（WYP）和3个养殖群体——朝鲜半岛群体的养殖一代（FKN），黄海1号（HH1）和即抗98（JK98）进行杂交试验的研究，结果表明JK98 (♀)  WKN (♂)组合在存活率方面最高，其余的依次为WYP (♀) WKN (♂)，WKN (♀) WYP (♂)，FKN (♀)HH1 (♂) 和 WYP (♀) FKN (♂)。而在体重方面FKN(♀)  HH1(♂)组合最高，其余的依次为WKN (♀) WYP (♂)，WYP (♀) WKN (♂)，WYP (♀)FKN (♂) 和 JK98 (♀)WKN (♂)。在所有生长性状方面，JK98 (♀)  WKN (♂)在5个组合中是最低的。方差检测结果表明，TL、CL、HST、LL和BW这5个指标在不同组合间存在差异，而其他指标不存在差异。多重比较结果发现JK98 (♀)WKN (♂)组合的TL与其他组合间差异极显著，HST指标与WKN (♀) WYP (♂)，FKN(♀)  HH1(♂)和 WYP (♀)  WKN (♂)这3个组合差异显著，BW指标与WKN (♀) WYP (♂) 和 FKN(♀)  HH1(♂)差异显著。 4. 通过人工授精的方法建立了中国对虾21个半同胞家系，测量了中国对虾21个半同胞（46个全同胞）家系的TL、CL、CW、HST、WST、第1腹节长（FL）、第6腹节长（LL）。利用MTDFREML软件得到生长性状遗传力在0.15~0.35之间，属于中度遗传力范围。TL的遗传力为0.34±0.071，CL的为0.30±0.070，CW为0.35±0.077，WST为0.33±0.073，HST为0.33±0.073，FL的最低为0.15±0.044，LL的为0.24±0.059。各个性状间表现出高的正相关，其中CW和TL以及HST的遗传相关最大，FL和WST的遗传相关最小。通过以上杂交育种和选择育种的研究，认为单纯的依靠杂交育种来改善中国对虾的育种工作可能具有一定的局限性。所以在实际的育种过程中，以中国对虾不同群体的杂交后代作为基础群体，并以此为基础进行系统的选择育种应该具有更大的潜力。 5. 本试验利用中国对虾F2群体和AFLP分子标记技术进行了遗传连锁图谱的构建。利用55对AFLP引物组合对F2家系的110个个体进行了研究，结果检测到532个符合作图策略的AFLP标记。利用卡方检验检测分离位点是否符合孟德尔分离定律。对于符合3：1比例的分离位点利用F2自交模型构建性别平均连锁图谱，对于符合1：1比例的分离位点利用拟测交理论分别构建中国对虾的雌性和雄性遗传连锁图谱。雌性、雄性和性别平均遗传图谱分别有28、35和44个连锁群，图谱实际长度分别为1090、1617和1772.1 cM。中国对虾遗传连锁图谱估计基因组长度为2420 cM，符合与人类基因组相比的对虾类基因组长度。中国对虾雄性遗传连锁图谱比雌性遗传连锁图谱长32.6%，这可能说明中国对虾不同性别存在不同的重组率。通过皮尔逊相关系数检测认为AFLP标记在中国对虾图谱上分布均匀。本文利用AFLP标记构建的中国对虾遗传连锁图谱为中国对虾基因组研究和遗传改良提供一定的基础，同时也应该开发微卫星等共显性标记，为遗传连锁图谱的整合提供条件。
其他摘要	The paper presented the appropriate breeding methods and the genetic linkage maps in Fenneropenaeus chinensis. The crossbreeding of F. chinensis and genetic parameters estimates were carried out. The genetic linkage maps were constructed. The results would provide theoretic basis for indirect and early breeding for F. chinensis. 1. In order to learn growth characteristics of different geographical populations, intra- and intercrosses of Rushan (WYP) and Korea (WKN) populations were produced. The body length (TL), the carapace length (CL), carapace width (CW), height of the second and third abdominal segment (HST), width of the second and third abdominal segment (WST), body weight (BW) and survival were measured. The heterosis of hybrids was computed, variances were analyzed, and multi-averages were compared. The results indicated that there existed different extent heterosis (4.37%~23.96%) for growth except survival in hybrids at the third month. The offspring of WYP♀ × WKN♂ had the highest heterosis at the third month, the less was WKN♀ × WYP♂, the least was WKN. The effects of morphological trait on body weight of F. chinensis were analyzed in this study.The multiple regression equation describing the body weight is: Y = －2.056 + 0.03X1 + 0.076X2 + 0.078X3 + 0.033X4 + 0.043X5. 2. At the fourth month, six indexes of growth trait and viability showed a range of heterosis, ranging from 0.514% to 14.950%. The heterosis of WYP♀ × WKN♂ was higher than that of the WKN♀ × WYP♂ in all indexes. At the fifth month after post-larvae, six indexes of growth trait and viability ranged from -9.000% to 19.090%. The negative heterosis was observed in CL, HST and viability. The heterosis of WKN♀ × WYP♂ tended generally to increase as the age of the Chinese shrimp increased while the heterosis of WYP♀ × WKN♂ tended to decrease. The results indicated that the viability of reciprocal hybrids were not significantly different from their parents. The result of ANOVA indicated that the hybrids were significantly different in WST and BW at the fourth month. The multiple comparisons of LSD test indicated that the hybrids of WYP♀ × WKN♂ were significantly different from their parents in BW. The hybrids of WYP♀ × WKN♂ were significantly different from the other three combinations in WST. At the fifth months, the hybrids had significant difference in BW while other traits showed no significant differences from their parents. The multiple comparisons of LSD test indicated that the hybrids of WKN♀ × WYP♂ were significantly different from the WKN parents in BW. 3. The crosses of two wild populations and three farmed populations were produced. The results indicated the survival of JK98 (♀)  WKN (♂) was the highest, followed by WYP (♀)  WKN (♂), WKN (♀)  WYP (♂), FKN (♀)  HH1 (♂) and WYP (♀)  FKN (♂). While the BW of FKN(♀)  HH1(♂) was the highest, followed by WKN (♀)  WYP (♂), WYP (♀)  WKN (♂), WYP (♀)  FKN (♂) and JK98 (♀)  WKN (♂). The TL had the same ranking as the BW. All growth traits in hybrids of JK98 (♀)  WKN (♂) were lowest among all combinations. The hybrids had significant difference in TL, CL, HST, LL and BW. The hybrids were not significant difference in other growth traits and survival rate. The results of Duncan’s Multiple Range Test were that the TL and CL of JK98 (♀)  WKN (♂) were significant difference from the other combinations, HST difference from the combination of the WKN (♀)  WYP (♂), FKN(♀)  HH1(♂) and WYP (♀)  WKN (♂) and the BW difference from WKN (♀)  WYP (♂) and FKN(♀)  HH1(♂). 4. Twenty-one half-sib groups of the Chinese shrimp were obtained using artificially insemination of two or three females mated each male. Seven growth traits of Chinese shrimp were measured. The heritability was estimated through multiple-trait derivative-free restricted maximum likelihood (MTDFREML). The heritability estimates for all growth traits calculated were significantly different from zero, ranging from 0.15±0.044 to 0.35±0.077. The heritability of TL was 0.34±0.071, CL 0.30±0.070, CW 0.35±0.077, WST 0.33±0.073, HST 0.33±0.073, FL 0.15±0.044 and LL 0.24±0.059, respectively. Genetic correlations were high between growth traits. The genetic correlation between CW and TL was the highest (1.0±0.007), between FL and WST the lowest (0.82±0.085). The crossing scheme and selective breeding indicated the exploitation of heterosis through single crossbreeding in Chinese shrimp was of limiting utility in practical commercial shrimp aquaculture in spite of the potential of significant heterosis. The crossbreeding of different populations can be applied in the establishment of base populations. The systemic selection program based on additive genetic performance may be more effective improvement than crossbreeding. 5. The linkage maps were constructed using a F2 population and amplified fragment length polymorphic (AFLP) markers. Fifty-five selected AFLP primer combinations produced 532 AFLP markers in 110 progeny that were fitted for map strategy. From them, the markers in 3:1 segregating ratios were analyzed using F2 intercross model strategy for the common linkage map, the markers in 1:1 ratio using the pseudo-testcross strategy. The separate male and female and common maps for 110 individuals were constructed. The female map included 103 markers which formed 28 linkage groups covering a total length of 1090 cM. In the male map, 144 markers were used to analyzed and grouped to 35 linkage groups covering a total distance of 1617cM, 10 makers unlinked. The common map, 216 markers were used to analyze and group to 44 linkage groups covering a total distance of 1772.1cM, 2 makers unlinked. An average estimated genome size for Chinese shrimp was 2420 cM, which were consistent with the relative size of the penaeid genome compared to the human genome. In Chinese shrimp, the male map was 32.6% larger than the male map, which may reflect sex-specific recombination rates in Chinese shrimp. The distribution of AFLP markers is relatively even in chromosomes of map. The linkage analysis presented in this paper had provided the first insight into the level of polymorphism and genetic variation of Chinese shrimp using AFLP. There has been a further effort to develop additional informative multiplexed microsatellite markers system to establish more saturated linkage map and carry out comparative genomic mapping.
页数	128
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/671
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	田燚. 中国对虾育种方法研究与遗传连锁图谱的构建[D]. 海洋研究所. 中国科学院海洋研究所,2007.

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