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绿色荧光蛋白在α-1,3半乳糖基转移酶敲除猪组织器官的表达分析 绿色荧光蛋白在α-1,3半乳糖基转移酶敲除猪组织器官的表达分析
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-08-12 DOI: 10.16288/J.YCZZ.15-160
李智方, 冯冲, 纪慧丽, 石宁宁, 宋小凤, 赵勤丽, 龙川, 潘登科, 杨小淦
The pig is an ideal source to provide organs because its organ size and physiology are similar to humans. However, an acute rejection will ensue after pig-to-human xenotransplantation. The α-1,3 galactosyltransferase gene knockout (GTKO) pigs were generated in recent years, and could solve the problem of hyperacute rejection. But due to lack of reporting genes, the rejection status of cells and organs post pig-to-human xenotransplantation cannot be visualized. In this study, we introduced the enhanced green fluorescent protein (EGFP) gene driven by the CAG promoter into GTKO porcine ear fibroblasts. Then we produced transgenic pigs expressing the EGFP gene by nuclear transfer technology. Expression levels of EGFP in different tissues and organs of the cloned pig were investigated by Nightsea DFP-1 Fluorescent Protein Flashlight, fluorescence microscope and quantitative PCR assays. The results showed that the protein and transcript of EGFP were expressed in all tissues and organs of the GTKO pig, but the expression was weak in the liver and central nervous system. In conclusion, we have successfully produced the transgenic GTKO pigs expressing EGFP in all tested tissues and organs, which builds up a good basis to track transplanted cells or tissues.
猪是提供器官的理想来源,因为它的器官大小和生理机能与人类相似。然而,猪与人异种移植后会出现急性排斥反应。α-1,3半乳糖转移酶基因敲除(GTKO)猪是近年来出现的,可以解决超急性排斥反应问题。但由于缺乏报告基因,细胞和器官在猪到人异种移植后的排斥状态无法可视化。本研究将CAG启动子驱动的增强型绿色荧光蛋白(EGFP)基因导入GTKO猪耳成纤维细胞。然后通过核移植技术制备表达EGFP基因的转基因猪。利用Nightsea DFP-1荧光蛋白手电筒、荧光显微镜和定量PCR检测EGFP在克隆猪不同组织器官中的表达水平。结果表明,EGFP蛋白及其转录本在GTKO猪的所有组织器官中均有表达,但在肝脏和中枢神经系统中表达较弱。综上所述,我们成功培育出在所有组织器官中表达EGFP的转基因GTKO猪,为追踪移植细胞或组织奠定了良好的基础。
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引用次数: 0
Wild Estonian and Russian sea trout (Salmo trutta) in Finnish coastal sea trout catches: results of genetic mixed-stock analysis 芬兰沿海捕捞的野生爱沙尼亚和俄罗斯海鳟(Salmo trutta):遗传混合种群分析的结果
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00070
Marja-Liisa Koljonen, Riho Gross, Jarmo Koskiniemi

For responsible fisheries management of threatened species, it is essential to know the composition of catches and the extent to which fisheries exploit weak wild populations. The threatened Estonian, Finnish and Russian sea trout populations in the Gulf of Finland are targets of mixed-stock fisheries. The fish may originate from rivers with varying production capacities, from different countries, and they may also have either a wild or hatchery origin. In order to resolve the composition of Finnish coastal sea trout catches, we created a standardized baseline dataset of 15 DNA microsatellite loci for 59 sea trout populations around the Gulf of Finland and tested its resolution for mixed-stock analysis of 1372 captured fish. The baseline dataset provided sufficient resolution for reliable mixture analysis at regional group level, and also for most of the individual rivers stocks. The majority (76–80%) of the total catch originated from Finnish sea trout populations, 6–9% came from Russian and 12–15% from Estonian populations. Nearly all Finnish trout in the catch were of hatchery origin, while the Russian and Estonian trout were mostly of wild origin. The proportion of fish in the Finnish catches that originated from rivers with natural production was at least one fifth (22%, 19–23%). Two different spotting patterns were observed among the captured trout, with a small and sparsely spotted form being markedly more common among individuals of Russian (28%) and Estonian origin (22%) than among fish assigned to a Finnish origin (0.7%).

为了对受威胁物种进行负责任的渔业管理,必须了解渔获量的组成和渔业利用脆弱野生种群的程度。芬兰湾受到威胁的爱沙尼亚、芬兰和俄罗斯海鳟鱼种群是混合种群渔业的目标。这些鱼可能来自不同国家、具有不同生产能力的河流,也可能来自野生或孵化场。为了解决芬兰沿海海鳟鱼捕捞的组成,我们为芬兰湾周围的59个海鳟鱼种群创建了15个DNA微卫星位点的标准化基线数据集,并测试了其对1372条捕获鱼的混合种群分析的分辨率。基线数据集提供了足够的分辨率,可以在区域群水平上进行可靠的混合分析,也可以对大多数单个河流种群进行分析。总捕捞量的大多数(76-80%)来自芬兰海鳟种群,6-9%来自俄罗斯,12-15%来自爱沙尼亚种群。几乎所有捕获的芬兰鳟鱼都来自孵化场,而俄罗斯和爱沙尼亚的鳟鱼大多来自野生。在芬兰的渔获量中,来自自然生产的河流的鱼的比例至少为五分之一(22%,19-23%)。在捕获的鳟鱼中观察到两种不同的斑点模式,小而稀疏的斑点形式在俄罗斯(28%)和爱沙尼亚(22%)的个体中明显比在芬兰(0.7%)的个体中更常见。
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引用次数: 29
An improved MALDI-TOF mass spectrometry procedure and a novel DNA marker for identifying over-expressed Bx7 glutenin protein subunit in wheat 一种改进的MALDI-TOF质谱方法和一种鉴定小麦过表达Bx7谷蛋白亚基的新型DNA标记
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00069
Ke Wang, Shahidul Islam, Junhong Ma, Masood Anwar, Jing Chen, Yueming Yan, Rudi Appels, Wujun Ma

Wheat bread-making quality is mainly determined by glutenin proteins in the grain, which exist in a wide range of variable alleles with differential influence on processing attributes. A recently identified allele, Bx7 over-expression (Bx7oe), has been showing highly significant positive effects on wheat dough strength over the normally expressed Bx7 allele. SDS-PAGE and normal RP-HPLC procedures failed to separate the two alleles. In the current study, an extensively optimised MALDI-TOF based procedure and a refined DNA based marker for efficiently differentiating Bx7oe from normal Bx7 allele were established. Results indicated that the MALDI-TOF procedure is cost effective, high throughput, and proven reliable, while the refined PCR marker only amplifies Bx7oe allele, a clear advantage over the previously developed codominant marker.

小麦面包的品质主要由籽粒中的谷蛋白决定,谷蛋白存在于广泛的可变等位基因中,对加工属性有不同的影响。最近发现的一个等位基因Bx7过表达(Bx7oe)对小麦面团强度的影响显著高于正常表达的Bx7等位基因。SDS-PAGE和常规反相高效液相色谱法无法分离这两个等位基因。在目前的研究中,我们建立了一个广泛优化的基于MALDI-TOF的程序和一个改进的基于DNA的标记,用于有效区分Bx7oe和正常Bx7等位基因。结果表明,MALDI-TOF方法具有成本效益、高通量和可靠性,而改进的PCR标记仅扩增Bx7oe等位基因,与先前开发的共显性标记相比具有明显优势。
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引用次数: 4
Genetic polymorphism of 15 Y chromosomal STR loci and haplotypes of Henan Han population 河南汉族人群15个Y染色体STR位点的遗传多态性及单倍型分析
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00067
Xiu-Hua Zhang, Wen-Bo Zhang, Xue-Hui Fan

We studied and established a DNA database of 15 Y-STRs (DYS438, DYS446, DYS391, DYS390, DYS458, DYS534, DYS426, DYS626, DYS504, DYS505, DYS576, DYS532, DYS594, DYS522, DYS540) in a population sample of 102 unrelated, healthy, male individuals of Henan Han population. Allelic frequencies and statistical parameters of Han population were calculated. Totally 90 alleles were observed, with the corresponding allelic frequencies ranging from 0.0098 to 0.9020. 102 haplotypes were found in the studied group, the haplotype diversity for 15 Y-STR loci was 1. The results of present study were valuable for human identification and paternity tests routine forensic applications in the region.

研究并建立了河南省汉族人群102例无亲缘关系健康男性个体的15个Y-STRs (DYS438、DYS446、DYS391、DYS390、DYS458、DYS534、DYS426、DYS626、DYS504、DYS505、DYS576、DYS532、DYS594、DYS522、DYS540) DNA数据库。计算汉族人群的等位基因频率和统计参数。共检测到90个等位基因,对应的等位基因频率范围为0.0098 ~ 0.9020。共发现102个单倍型,其中15个Y-STR基因座的单倍型多样性为1。本研究的结果对人类鉴定和亲子鉴定在该地区的常规法医应用具有价值。
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引用次数: 3
Deletion mapping in the Enhancer of split complex 分裂复合体增强子中的删除映射
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00065
Elisa Wurmbach, Anette Preiss

The Enhancer of split complex [E(spl)-C] comprises twelve genes of different classes. Seven genes encode proteins of with a basic-helix-loop-helix-orange (bHLH-O) domain that function as transcriptional repressors and serve as effectors of the Notch signalling pathway. They have been named E(spl)m8-, m7-, m5-, m3-, mβ-, mγ- and mδ-HLH. Four genes, E(spl)m6-, m4-, m2- and mα-BFM are intermingled and encode Notch repressor proteins of the Bearded-family (BFM). The complex is split by a single gene of unrelated function, encoding a Kazal-type protease inhibitor (Kaz-m1). All members within a family, bHLH-O or BFM, are very similar in structure and in function. In an attempt to generate specific mutants, we have mobilised P-element constructs residing next to E(spl)m7-HLH and E(spl)mγ-HLH, respectively. The resulting deletions were mapped molecularly and by cytology. Two small deletions affected only E(spl)m7-HLH and E(spl)mδ. The deficient flies were viable without apparent phenotype. Larger deletions, generated also by X-ray mutagenesis, uncover most of the E(spl)-C. The phenotypes of homozygous deficient embryos were analysed to characterize the respective loss of Notch signalling activity.

分裂复合体的增强子[E(spl)-C]由12个不同类别的基因组成。7个基因编码具有碱性螺旋-环-螺旋-橙色(bHLH-O)结构域的蛋白,其功能是转录抑制因子和Notch信号通路的效应器。它们被命名为E(spl)m8-、m7-、m5-、m3-、mβ-、mγ-和mδ-HLH。4个基因E(sp1)m6-、m4-、m2-和mα-BFM相互交织,编码BFM家族的Notch抑制蛋白。该复合体由一个功能不相关的基因分裂,该基因编码kazal型蛋白酶抑制剂(kazal -m1)。一个家族中的所有成员,bHLH-O或BFM,在结构和功能上都非常相似。为了产生特异性突变体,我们分别动员了位于E(spl)m7-HLH和E(spl)m - γ- hlh旁边的p元素构建体。由此产生的缺失被分子和细胞学定位。两个小的缺失只影响E(spl)m7-HLH和E(spl)mδ。有缺陷的果蝇可存活,无明显表型。较大的缺失,也由x射线诱变产生,揭示了大部分E(spl)-C。分析了纯合子缺陷胚胎的表型,以表征各自Notch信号活性的丧失。
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引用次数: 5
Association analysis of the GRM8 gene with schizophrenia in the Uygur Chinese population 维吾尔族人群中GRM8基因与精神分裂症的相关性分析
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00045
Lili Zhang, Xianjiang Zhong, Zhiguo An, Shuxian Han, Xiao Luo, Yongyong Shi, Qizhong Yi

GRM8 is a schizophrenia candidate gene that is also thought to be involved in the glutamate pathway, which is very important in the pathogenesis of schizophrenia. In this study, we aim to investigate the association between GRM8 and schizophrenia in the Uygur Chinese population.

Rs2237748 and rs2299472, located in the GRM8 gene, were selected for genotyping in a set of Uygur Chinese case-control samples, which included 723 cases and 561 controls, using TaqMan assays and capillary sequencing. The statistical analysis was carried out using the online software program SHEsis, and a meta-analysis was carried out to identify other relevant studies using Review Manager 5. We found that the rs2299472 genotype was significantly associated with schizophrenia (P = 0.015, P = 0.030, after Bonferroni correction). The frequency of the CC genotype was higher in the schizophrenic patients (P = 0.008), and the frequency of the AC genotype was lower (P = 0.008). Furthermore, the meta-analysis incorporating the previous and current studies also showed that rs2299472 is associated with schizophrenia. This study indicates that the GRM8 gene may play an important role in the pathogenesis of schizophrenia.

GRM8是一种精神分裂症候选基因,也被认为参与谷氨酸通路,这在精神分裂症的发病机制中非常重要。在这项研究中,我们的目的是调查GRM8与维吾尔族中国人精神分裂症之间的关系。选择GRM8基因中的Rs2237748和rs2299472,采用TaqMan法和毛细管测序方法,对维吾尔族人病例对照样本(723例和561例对照)进行基因分型。使用在线软件SHEsis进行统计分析,并使用Review Manager 5进行meta分析以确定其他相关研究。我们发现rs2299472基因型与精神分裂症显著相关(经Bonferroni校正P = 0.015, P = 0.030)。精神分裂症患者CC基因型频率较高(P = 0.008), AC基因型频率较低(P = 0.008)。此外,结合以往和当前研究的荟萃分析也显示rs2299472与精神分裂症有关。本研究提示GRM8基因可能在精神分裂症发病机制中发挥重要作用。
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引用次数: 12
Analysis of the interaction between human RITA and Drosophila Suppressor of Hairless 人RITA与果蝇无毛抑制因子的相互作用分析
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00074
Birgit Brockmann, Helena Mastel, Franz Oswald, Dieter Maier

Notch signalling mediates intercellular communication, which is effected by the transcription factor CSL, an acronym for vertebrate CBF1/RBP-J, Drosophila Suppressor of Hairless [Su(H)] and C. elegans Lag1. Nuclear import of CBF1/RBP-J depends on co-activators and co-repressors, whereas the export relies on RITA. RITA is a tubulin and CBF1/RBP-J binding protein acting as a negative regulator of Notch signalling in vertebrates. RITA protein is highly conserved in eumatazoa, but no Drosophila homologue was yet identified. In this work, the activity of human RITA in the fly was addressed. To this end, we generated transgenic flies that allow a tissue specific induction of human RITA, which was demonstrated by Western blotting and in fly tissues. Unexpectedly, overexpression of RITA during fly development had little phenotypic consequences, even when overexpressed simultaneously with either Su(H) or the Notch antagonist Hairless. We demonstrate the in vivo binding of human RITA to Su(H) and to tubulin by co-immune precipitation. Moreover, RITA and tubulin co-localized to some degree in several Drosophila tissues. Overall our data show that human RITA, albeit binding to Drosophila Su(H) and tubulin, cannot influence the Notch signalling pathway in the fly, suggesting that a nuclear export mechanism of Su(H), if existent in Drosophila, does not depend on RITA.

Notch信号通路介导细胞间通讯,受转录因子CSL、脊椎动物CBF1/RBP-J、果蝇无毛抑制因子[Su(H)]和线虫Lag1的首字母缩写影响。CBF1/RBP-J的核进口依赖于共激活剂和共阻滞剂,而出口依赖于RITA。RITA是一种微管蛋白和CBF1/RBP-J结合蛋白,在脊椎动物中作为Notch信号的负调节因子。RITA蛋白在类人猿中高度保守,但尚未发现与果蝇同源的蛋白。本文研究了人RITA在蝇体内的活性。为此,我们通过Western blotting和果蝇组织实验证明,我们培育了能够组织特异性诱导人RITA的转基因果蝇。出乎意料的是,在果蝇发育过程中,即使与Su(H)或Notch拮抗剂Hairless同时过表达,RITA也几乎没有表型后果。我们通过共免疫沉淀证明了人RITA与Su(H)和微管蛋白在体内的结合。此外,RITA和微管蛋白在果蝇的几个组织中有一定程度的共定位。总的来说,我们的数据表明,人类的RITA虽然与果蝇的Su(H)和微管蛋白结合,但不能影响果蝇的Notch信号通路,这表明如果果蝇中存在Su(H)的核输出机制,则不依赖于RITA。
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引用次数: 7
A tale of two papers 两张纸的故事
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00073
Anssi Saura

Two papers published in HEREDITAS between 1921 and 1939 show how the attitude towards race biology changed in the course of the interwar period in the Nordic countries. In the early 1920s race biology was seen to constitute a legitimate science. Ordinary human genetics prevailed, however, over race biology already in the very beginning on the pages of HEREDITAS. Population thinking was introduced into the study of human heredity around the year 1930. It effectively contradicted the concept of the race. Interestingly, HEREDITAS does not carry a single paper on eugenics and sterilization. In 1939 we see a final repudiation of the doctrines on race. Times had changed and the National Socialists had usurped the doctrines of race in Germany.

1921年至1939年间发表在《遗传》杂志上的两篇论文显示了北欧国家在两次世界大战之间对种族生物学的态度是如何变化的。在20世纪20年代早期,种族生物学被认为是一门合法的科学。然而,在《遗传》杂志的一开始,普通的人类遗传学就已经战胜了种族生物学。大约在1930年,人口思维被引入到人类遗传的研究中。它有效地与种族的概念相矛盾。有趣的是,HEREDITAS没有一篇关于优生学和绝育的论文。在1939年,我们看到对种族理论的最终否定。时代变了,国家社会主义者篡夺了德国的种族理论。
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引用次数: 3
Strange history: the fall of Rome explained in Hereditas 奇怪的历史:《赫里达斯传》解释了罗马的灭亡
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00080
Bengt O. Bengtsson

In 1921 Hereditas published an article on the fall of Rome written by the famous classical scholar Martin P:son Nilsson. Why was a paper on this unexpected topic printed in the newly founded journal? To Nilsson, the demise of the Roman Empire was explained by the “bastardization” occurring between “races” from different parts of the realm. Offspring from mixed couples were of a less stable “type” than their parents, due to the breaking up by recombination of the original hereditary dispositions, which led to a general loss of competence to rule and govern. Thus, the “hardness” of human genes, together with their recombination, was – according to Nilsson – the main cause of the fall of Rome.

Nilsson's argument is not particularly convincingly presented. Human “races” are taken to have the same genetic structure as inbred crop strains, and Nilsson believes in a metaphysical unity between the individual and the race to which it belongs. However, in my view, Martin P:son Nilsson and his friend Herman Nilsson-Ehle had wider aims with the article than to explain a historical event. The article can be read as indicating strong support from the classical human sciences to the ambitious new science of genetics. Support is also transferred from genetics to the conservative worldview, where the immutability and inflexibility of the Mendelian genes are used to strengthen the wish for greater stability in politics and life. The strange article in Hereditas can, thus, be read as an early instance in the – still ongoing – tug-of-war between the conservative and the liberal ideological poles over how genetic results best are socially interpreted.

1921年,赫雷迪塔斯发表了一篇关于罗马衰亡的文章,作者是著名的古典学者马丁·P·尼尔森。为什么一篇关于这个意想不到的话题的论文会刊登在新创办的杂志上?对尼尔森来说,罗马帝国的灭亡可以用来自不同地区的“种族”之间的“私生子化”来解释。混血夫妇的后代比他们的父母的“类型”更不稳定,因为原始遗传倾向的重组导致了分裂,这导致了统治和管理能力的普遍丧失。因此,根据尼尔森的说法,人类基因的“硬度”,以及它们的重组,是罗马灭亡的主要原因。尼尔森的论点并没有特别令人信服。人类“种族”被认为与近亲繁殖的作物品种具有相同的遗传结构,尼尔森相信个人与其所属种族之间存在形而上学的统一。然而,在我看来,Martin P:son Nilsson和他的朋友Herman Nilsson- ehle写这篇文章的目的比解释一个历史事件更广泛。这篇文章可以被解读为表明了古典人文科学对雄心勃勃的基因学新科学的大力支持。遗传学的支持也转移到保守的世界观,孟德尔基因的不变性和不灵活性被用来加强对政治和生活更稳定的愿望。因此,《遗传》杂志上的这篇奇怪的文章可以被解读为保守派和自由派意识形态两极之间关于基因结果如何得到最好的社会解释的拉锯战的早期例子,这场拉锯战至今仍在进行。
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引用次数: 3
Genetic diversity and population structure of leafy kale and Brassica rupestris Raf. in south Italy 叶甘蓝和芸苔的遗传多样性和群体结构。在意大利南部
IF 2.7 3区 生物学 Q3 GENETICS & HEREDITY Pub Date : 2015-01-14 DOI: 10.1111/hrd2.00058
Lorenzo Maggioni, Roland von Bothmer, Gert Poulsen, Ferdinando Branca, Rikke Bagger Jørgensen

Local varieties of leafy kales (Brassica oleracea L.) are grown in home gardens in Calabria and Sicily for self-consumption, in the same area where the wild relative Brassica rupestris Raf. also grows. With the use of AFLP markers, comparisons were made of the genetic diversity and population structure of ten wild and 22 cultivated populations, as well as of a hybrid population and of four commercial cultivars of different B. oleracea crops. The level of genetic diversity was higher in leafy kales than in wild populations and this diversity was mainly distributed within populations. Wild populations remained distinct from cultivated material. Additionally, most wild populations were distinctively isolated from each other. On the other hand, it was not possible to molecularly distinguish even geographically distant leafy kale populations from each other or from different B. oleracea crops. It was possible to detect inter-crossing between leafy kales and B. rupestris. Findings from this study illustrate the existing level of genetic diversity in the B. oleracea gene pool. Individual populations (either wild or leafy kales) with higher levels of genetic diversity have been identified and suggestions are given for an informed conservation strategy. Domestication hypotheses are also discussed.

在卡拉布里亚和西西里岛,当地品种的叶甘蓝(芸苔甘蓝L.)在家庭花园里种植,供自己食用,在同一地区,野生近亲芸苔(Brassica rupestris Raf)。也在增长。利用AFLP标记对10个野生群体和22个栽培群体、1个杂交种群体和4个商业品种的遗传多样性和群体结构进行了比较。叶甘蓝的遗传多样性水平高于野生居群,且主要分布在居群内。野生种群仍然不同于栽培材料。此外,大多数野生种群彼此之间存在明显的隔离。另一方面,即使在地理上相距遥远的羽衣甘蓝群体之间或不同的甘蓝作物之间,也不可能从分子上区分开来。叶羽衣甘蓝与鲁氏白僵菌之间存在交叉杂交。本研究结果说明了甘蓝基因库中存在的遗传多样性水平。已经确定了具有较高遗传多样性水平的单个种群(无论是野生还是叶甘蓝),并提出了明智的保护策略建议。本文还讨论了驯化假说。
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引用次数: 19
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