青藏高原沙蜥的进化研究
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摘要
青藏高原面积约240 Km~2,大部分地区海拔在4000-5000 m。上新世以来,青藏高原隆升塑造了高原内部和周遍地区独特的异质性自然环境,这对高原地区生物多样性的形成产生了重要影响,使高原成为地球中低纬度生物多样性高分布地区与生物多样性研究的模式地之一。
     沙蜥是青藏高原最为醒目的爬行动物,几乎遍布整个高原,种群数量丰富,是研究青臧高原动物进化生物学的好材料。以青臧高原隆升,环境演变为背景,研究高原特有的沙蜥的进化生物学具有无可替代的科学理论价值。本论文选择青藏高原上沙蜥属的物种为研究对象,从形态学、生态学、分子生物学、生态遗传学、生物地理学、系统地理学、系统发生和分类学等多个学科交叉研究其进化。
     在种内,青海沙蜥个体随海拔升高变小,四肢相对长度增长,与Bergmann规律和Allen规律不相符。青海沙蜥个体大小的变异受与海拔梯度相关的环境因素的作用,如高海拔低温、低氧、短的活动季节以及低的食物可利用性等。在高海拔动物有氧代谢活动受限的情况下,相对增加四肢的生长可能与沙蜥增强活动能力有关。
     在种间,青海沙蜥个体比低海拔荒漠种组大,符合Bergmann规律。青海沙蜥尾椎骨数目显著少于低海拔荒漠种组的尾椎骨数。此外,与低海拔相比,高海拔青海沙蜥种群相对尾长缩短、平均尾椎骨数目减少。这说明尾长变异符合Allen规律,尾椎骨数变异不符合Jordan规律。其种间或种内形态的变异主要受到了遗传和系统发生的影响。
     此外,分布于寒冷干燥环境下的西藏沙蜥腹部有黑斑,且多具棕红色斑,而分布于较温暖和湿润地区的西藏沙蜥不具这一特点,腹部体色特征的变异不符合Gloger规律。
     青海沙蜥高海拔种群雌性产小窝大仔,具有较小的窝仔数变异系数、低的窝仔投入,但给每个小仔的投入高。青海沙蜥种群生殖对策从低海拔的r-对策逐渐过渡到高海拔的K-对策。
     青海沙蜥的遗传多样性随海拔升高下降,随温度升高而升高,与降雨成负相关,在温度和降雨稳定地区遗传多样性较高。说明在高海拔低温、缺氧、繁殖季节短、太阳辐射强烈以及多变的环境作用下,只有少数遗传型才能适应。这种遗传多样性的变异可能是高海拔个体大小和K-对策选择的遗传基础。
     青藏高原沙蜥mtDNA的进化并未趋于饱和,分子进化受中性选择作用,并且受到自然选择压力。与其它几种沙蜥相比,分布海拔最高的红尾沙蜥碱基异义替换与同义替换比值最大,其mtDNA仍处于强烈适应进化中。
     青藏高原沙蜥,包括红尾沙蜥、贵德沙蜥和青海沙蜥,在系统树上共同组成一个单系,分歧时间发生在5.06±0.68百万年前(Million years ago,Mya)。大部分遗传变异发生在分支或地区之间,表明高原沙蜥各大分支之间基因交流长期中断。青海沙蜥系统地理格局的形成主要受到上新世和更新世高原三个主要隆升时期,即青藏运动A幕(3.4 Mya)、B幕(2.5 Mya)和C幕(1.7 Mya)的影响。在这个时期发生的高原整体隆升和山脉抬升、昆黄运动、共和运动以及湖泊演变,形成了高原内部迥异的地质地貌,限制了沙蜥的基因交流,塑造了高原沙蜥的系统地理结构和历史种群动态。
     青藏高原沙蜥系统树上有三个古老分支,在地理上相当于西藏沙蜥和泽当沙蜥分布区。两个分支分歧早于贵德沙蜥,其中最早从祖先分歧出来的一支分布于藏南谷地,另外一支分布于藏南谷地以西喜马拉雅山和冈底斯山之间的雅鲁藏布江、马泉河流域。还有一支组成贵德沙蜥的姐妹支,分布于西藏阿里地区,并且与分布于藏南谷地的分支分布区重叠。重叠分布的两大分支样本形态非常相似,并且与重叠区中的一个分支在阿里地区分布的样本形态差异明显,表明重叠区两分支祖先二次相遇后很可能发生了基因的相互渗透,或是受到相同环境下趋同进化的影响而使形态相似。
     贵德沙蜥进化分支和青海沙蜥柴达木分支在共和沙珠玉地区重叠分布,重叠区相当于共和古湖。受共和运动的作用,共和古湖在0.15 Mya迅速消失,演变为共和盆地,青海沙蜥和贵德沙蜥祖先得以进入共和盆地二次相遇,并发生杂交。
     以分子系统地理、系统分类原理和方法,以及遗传学证据厘定了青藏高原沙蜥属的种和种下分类。南疆沙蜥(P.forsythii)、贵德沙蜥(P.putjatia)、西藏沙蜥(P.theobaldi)、泽当沙蜥(P.zetangensis)和东方沙蜥(P.orientalis)为单型种,红尾沙蜥(P.erythrurus)和青海沙蜥(P.vlangalii)为多型种。红尾沙蜥分化为指名亚种(P.erythrurus erythrurus)和莫云亚种(P.erythrurus parva),两亚种由唐古拉山隔离。青海沙蜥分化为指名亚种(P.vlanglaii vlangalii)、普氏亚种(P.vlangalii pylzowi)与南山亚种(P.vlangalii nanschanica),它们之间的地理分布不连续,基因交流间断,独立进化。
     总之,晚中新世以来青藏高原及所属山脉的抬升、更新世的冰川作用、昆黄运动、共和运动以及湖泊的演替决定了青臧高原沙蜥属的分子进化、物种形成、亚种分化与种群系统地理结构。高原的抬升所造就的高海拔低温、缺氧、短的活动季节和低的食物可利用性的环境,决定了青海沙蜥形态和生活史对策。
The Qinghai-Xizang(Tibetan)Plateau is an immense upland(approximately 2.4 million km~2),averaging more than 4000 m above sea level in altitude.Environmental heterogeneity in internal and adjacent areas of Tibetan Plateau had been greatly shaped by the uplifting of plateau since Pliocene,which had great impact on the formation of biological diversity in the plateau and drove the plateau overlapping with the southwestern China mountains' biodiversity hotspot.Moreover,the plateau is becoming a model region for the researches of biodiversity.
     Distributing almost the whole Tibetan Plateau,Phrynocephalus lizards have been paid attention and their tremendous populations provide good model organisms for analyzing evolutionary biology.Because of continuing uplifting and environmental changing,Tibetan Plateau had unique environmental diversity and biological diversity. Studies on those endemic species have great and irreplaceable theoretical value.As model organisms,a number of researches concerning evolutionary biology on Phrynocephalus in Tibetan Plateau have been done,including morphology,ecology, molecular biology,ecological genetics,phylogeography,phylogeny and taxonomy.
     We investigated the evolution on morphology and reproductive characteristics in P.vlangalii along elevational and climatic gradients.The morphological variation pattern follows the converse to Bergmann's rule and it suggests that this body size pattern is driven by temperature,air pressure,length of the activity season and available food resources.However,P.vlangalii has shorter relative limb length which mismatches the Alien's rule and it is suggested to strengthen the ability of activity in high altitude with hypoxia environment.
     Besides these morphological variations mentioned above,P.vlangalii has fewer tail vertebrae and larger body size than oviparous Phrynocephalus lizard in low elevations,which indicates that these variations among species of Phrynocephalus are affected mainly by the genetic and phylogenetic factors.As a result,these variations match the Bergmann's rule and mismatch the Jordan's rule.However,P.theobaldi in colder and more arid environments was often found with brown-red patch and/or black plaque in abdomen,but could not be found in warmer and wetter environments, e.g.,valleys in southern Tibet.
     It could be concluded that the reproductive strategies of P.vlangalii fit into the common pattern of animals in higher elevation,which have smaller clutches of larger offspring and lower reproductive output.At higher altitudinal environments,the lizards have lower coefficient of litter size variation.The decreased litter size cline along elevational gradients might also be correlated with more anatomical constraints at higher elevations.This strategy of females showed the shifting from r-strategy to K-strategy along with increasing altitude.
     The genetic diversity of P.vlangalii decreased with increasing elevation, increasing rainfall and decreasing environmental temperature.There was a higher genetic diversity in temperature and rainfall stable regions.These were similar with the results of P.vlangalii' evolution on morphology and reproduction,which suggests that fewer genotypes of P.vlangalii are selected in higher elevation and extreme environments,e.g.lower temperature,hypoxia,stronger solar radiation.
     The test on nucleotide substitution indicates that the evolution of mtDNA has not reached saturation.Moreover,P.erythrurus has the highest nucleotide nonsynonymous substitution/synonymous substitution value in Phrynocephalus lizards in Tibet.The results indicate that P.erythrurus is still in the process of adaptive evolution under higher altitudinal environments.
     Phylogenetic analyses revealed that P.vlangalii and two other proposed species (P.erythrurus and P.putjatia)together form a monophyletic mtDNA clade,which in contrast with previous studies does not include P.theobaldi and P.zetangensis.The main P.vlangalli clade comprises seven well-supported lineages that correspond to distinct geographical areas with little or no overlap,and share a most recent common ancestor at 5.06±0.68 million years ago(mya).Analyses of molecular variance indicated that most of the observed genetic variations occurred among populations/regions,which implys the long-term interruption of maternal gene flow. A combined approach based on tests of population expansion,estimation of node dates,and significance tests on clade areas indicated that phylogeographical structuring has been primarily shaped by three main periods of plateau uplift during the Pliocene and Pleistocene,specifically 3.4 mya,2.5 mya and 1.7 mya.These periods corresponded to the appearance of several mountain ranges that formed physical barriers between lineages.Populations from the Qaidam Basin are shown to have undergone major demographic and range expansions in the early Pleistocene. This is consistent with colonization of areas previously covered by the huge Qaidam paleo-lake,which desiccated at this time.It was the role of uplifting of Tibetan Plateau(e.g.,Qingzang Movement),Kunlun-Yellow Movement,Gonghe Movement, the desiccating of paleo-lakes(e.g.,Qaidam and Gonghe paleolakes)that remolded the quite different geomorphology in Tibetan Plateau.These events shaped current patterns of diversity.
     Analyses on phylogeography of Phrynocephalus in Tibetan Plateau discovered two older evolutionary lineages than the clade corresponding to P.putjatia.One earlier diverged clade corresponding to valleys in southern Tibet,and another corresponding to the west of southern Tibet valleys between Himalayas and Gangdise mountains.The third but latest diverged clade of P.theobaldi nested as a sister clade with P.putjatia,its distributions not only covered the Ali area in Tibet,but reached into Galleys in southern Tibet.Interestingly,our results showed that the earliest diverged clade was completely overlapped with the latest diverged clade in the valleys of southern Tibet.There were obviously morphological differences between specimens from valleys in southern Tibet and from Ali region in western Tibet. Moreover,we propose that lizards with similar morphology of these two geographically overlapped clades might have been undergoing introgression since second contact occurred.However,besides hybridization on driving morphological convergence,it is still not safe to exclude the role of morphological convergent evolution under similar condition.
     MtDNA hybridization between P.vlangalii and P.putjatia was firstly reported in Phrynocephalus,which occurred in the sympatric region(Shazhuyu village,Gonghe county,Qinghai Plateau)and was considered as reciprocal introgression of mtDNA. The hybridization between the two lizard species was estimated at about 0.19-0.16 mya,which was coincided with the time of disappearance of the Gonghe paleo-lake at 0.15 mya,suggesting that the two species experienced second contact.Genetic introgression might occur after these two lizard species dispersed into the contact region when the Gonghe paleolake disappeared.
     We provided good evidence to resolve the molecular taxonomic chaos for Phrynocephalus complex in Tibetan Plateau with complete lineage sorting.There are five monotypic species including P.forsythii,P.putjatia,P.theobaldi,P.zetangensis, P.orientalis and two polytypic species including P.vlangalii and P.erythrurus in the whole plateau.P.erythrurus diverged into P.erythrurus erythrurus and P.erythrurus pylzowi because of intervening of Tanggula mountains.P.vlanglaii diverged into P. vlangalii vlangalii,P.vlangalii pylzowi and P.vlangalii nanschanica.These three subspecies have discontinuous distribution and have been undergone long-term interruption of gene flow.
     In summary,uplift of the Tibetan Plateau since Miocene is considered to be one of the most important geological events on shaping the diversity of Phrynocephalus. In more detail,many tectonic and geomorphical changing events e.g.,Qingzang Movement,Kun-Huang movement,Gonghe movement,evolution of paleo-lakes,had driven the formation of species and subspecies and shaped the geographical phylogenetic patterns of Phrynocephalus.Environmental heterogeneity,especially the low temperature,hypoxia,lower food availability and shorter seasonal activity, determined the evolutionary strategies on morphology and reproduction of P. vlangalii.
引文
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