丹顶鹤孵卵期行为比较和亲缘关系的研究
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摘要
丹顶鹤(Grus japonensis)属鹤形目、鹤科的大型涉禽,为国家Ⅰ级保护动物,是受国内外专家学者关注的珍稀野生动物资源。由于生态环境的改变、湿地的减少,使丹顶鹤赖以生存的栖息地日益丧失,导致丹顶鹤种群数量连年下降。当前丹顶鹤的保护工作已引起全世界的普遍关注。
     2007年4月在扎龙国家级自然保护区,采用瞬时扫描取样法对散养和野生丹顶鹤的孵卵期行为进行了观察。对动物个体各行为型发生频次进行了行为差异的显著性比较。
     利用扩增片段长度多态性(AFLP)初步建立了用于丹顶鹤遗传标记的新体系,筛选出28对选择性扩增引物,对5对丹顶鹤进行了多态性和亲缘关系分析,探讨了AFLP分子标记技术应用于丹顶鹤研究的可行性。为AFLP分子标记技术应用于丹顶鹤的亲缘关系鉴定提供了新方法,开辟了国内外对丹顶鹤行为遗传学研究的新途径。
     结果表明:
     1.散养丹顶鹤与野生丹顶鹤的孵卵期行为模式基本相同。在总体行为发生频次上,散养丹顶鹤的繁殖行为明显高于野生丹顶鹤,游走行为则低于野生丹顶鹤。行为日节律的比较表明,一天中各时间散养丹顶鹤的繁殖行为波动小于野生丹顶鹤。由于人为干扰,散养丹顶鹤的繁殖行为显著高于野生丹顶鹤的繁殖行为(P<0.05)。在个体间差异比较中,散养丹顶鹤与野生丹顶鹤在觅食表达上均存在显著差异(P<0.05),而散养丹顶鹤的警戒行为表达也存在显著差异(P<0.05)。
     2.丹顶鹤孵卵期的行为差异与其筑巢地点受干扰程度有很大关系,人为干扰是影响其行为的重要因素。散养鹤对人的存在并不敏感,使其在各行为的表达上与野生鹤有一定区别。人为干扰对野生鹤的繁殖行为有很大影响。
     3.利用28对引物对5对丹顶鹤进行了多态性和亲缘关系分析,共得到1114个扩增位点,其中多态性条带551条,多态性比例为49.5%。表明AFLP技术应用于丹顶鹤基因组分析可获得良好的效果。
     4.对AFLP数据进行聚类分析,构建了5对丹顶鹤的UPGMA聚类图,1、2号和3、4号鹤的亲缘关系较近,其余3对鹤亲缘关系较远。
Red-crowned crane(Grus japonensis)belongs to Gruiformes、Gruidae,it's a kind of large wading bird,level I of national protect animal.It's the rare wild animal resource that got attentions of scholars both at home and abroad.For the changes of ecological environment,marsh missing,the habitat for red-crowned crane to survive is disappearing day by day.This situation have made the population of red-crowned crane decreasing in consecutive years.Nowadays,the protection of red-crowned crane has been a cause for concern in world.
     By the method of continuous recording, a comparative study on the incubating behavior of semi-domestic and wild Red-crowned Crane (Grus japonensis) was made in April, 2007 in the Zhalong Nature Reserve.After getting the summation of every behavior frequency from animal individuals,we compared the behavior significant differences.
     Amplification fragment length polymorphisms(AFLP) is based on produce of the DNA fragment polymorphism by restriction enzyme and selective amplification to genomics.AFLP combines the stability and credibility of RFLP with the convenience and efficiency of RAPD, and can show high genetic polymorphism. AFLP of the red-crowned crane wasn't reported at home. This research preliminarily explored the AFLP on red-crowned crane , analysed the polymorphism and genetic relationship by 28 primer for AFLP selective amplification, discussed the feasibility about using AFLP in the research of the red-crowned crane.
     The research result as follows:
     1.The semi-domestic red-crowned crane and wild red-crowned crane basically have the same behavioral mode. In total behavior frequency, semi-domestic red-crowned crane showed more breeding behavior than the wild red-crowned crane obviously, and fewer walking behavior than the wild red-crowned crane.The behavior rhythm indicated that in one day the breeding behavior fluctuation of the semi-domestic red-crowned crane was smaller than the wild red-crowned crane.For the human activity,semi-domestic red-crowned crane exhibited more breeding behavior than wild red-crowned crane (P<0.05).In the individual comparison,there were significant differences in feeding behavior of semi-domestic and wild red-crowned crane (P<0.05),and significant difference in the guarding behavior of semi-domestic red-crowned crane (P<0.05).
     2.The differences of incubating behavior related the degree of disturbed at nesting sites,the influence of disturbance by humans is a important factor. Semi-domestic red-crowned crane is not sensitive to human,it makes differences in the expression behavior from wild red-crowned crane. The influence of disturbance by humans is strong to breeding behavior of wild red-crowned crane.
     3. By 28 primer for AFLP selective amplification, analysed the polymorphism and genetic relationship of 5 couples of red-crowned crane.Totally getting 1114 amplification sites,including 551 polymorphism sites,the percentage of polymorphism is 49.5%.It showed that by AFLP is a useful kind of molecular marker method in the research of genomics on red-crowned crane.
     4.By cluster analysis to AFLP data, An UPGMA tree of 5 couples of red-crowned crane had been made.No.1 and No.2 had closely relationship, No.3 and No.4 also had very closely relationship,The other cranes did not.
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