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Flexible DNA isolation procedure for different tree species as a convenient lab routine 灵活的不同树种DNA分离程序,作为方便的实验室常规
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0003
T. Bruegmann, M. Fladung, H. Schroeder
Abstract DNA isolation is a fundamental technique for all molecular biology laboratories. Depending on the plant species, DNA isolation can be challenging. In particular, adapted protocols rarely exist for tree species which are not used as standard model organisms. Here, we describe a flexible DNA isolation protocol that works for 59 tree species in a modular system. It is based on an ATMAB-containing extraction buffer to which proteinase K and/or boric acid are added, depending on the plant species. Subsequent purification steps include one or two precipitations with dichloromethane and, depending on the tree species, an optional sodium acetate precipitation. Using leaf material of a hybrid poplar clone from in vitro culture, it was determined that higher amounts of DNA could be isolated with this material than from field leaves. Starting from leaf material, DNA isolation for difficult cases was achieved with cambium or root tissue. This protocol was used to extract DNA for subsequent PCR amplification. Markers for cpDNA, mtDNA, and genomic DNA were used for standardized testing.
DNA分离是所有分子生物学实验室的一项基本技术。根据植物种类的不同,DNA分离可能具有挑战性。特别是,对于不用作标准模式生物的树种,很少存在适应的方案。在这里,我们描述了一个灵活的DNA分离协议,适用于59种树种的模块化系统。它是基于含有atmab的提取缓冲液,根据植物种类添加蛋白酶K和/或硼酸。随后的净化步骤包括用二氯甲烷进行一次或两次沉淀,并根据树种选择醋酸钠沉淀。利用离体培养的杂交杨树无性系叶片材料,测定了该材料比从田间叶片中分离出更多的DNA。从叶片材料开始,从形成层或根组织中分离DNA。该方案用于提取DNA用于随后的PCR扩增。采用cpDNA、mtDNA和基因组DNA标记进行标准化检测。
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引用次数: 4
Characterisation of nuclear microsatellite markers for Fraxinus excelsior L. and their transferability to six related species 黄曲霉核微卫星标记的特征及其在6个近缘种间的可转移性
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0008
Tina Hartung, K. Budde, O. Gailing
Abstract Common ash (Fraxinus excelsior L.) is an economically and ecologically important tree species in Europe acutely threatened by ash dieback. Here, we present a new set of Simple Sequence Repeat markers for F. excelsior and six related ash species based on published in silico designed primers. Forty SSR markers, mainly from genic regions, were tested for amplification and polymorphism and characterized in two ash populations in Germany. Transferability of polymorphic markers to six related Fraxinus species (F. angustifolia, F. ornus, F. quadrangulata, F. pennsylvanica, F. americana and F. biltmoreana) was also assessed. Eighteen markers, predominantly from genic regions including three markers closely linked to candidate genes for ash dieback, were successfully amplified and polymorphic in F. excelsior, of which between 10 to 17 were transferable to one of the six related species. High genetic diversity was found in the two ash populations (Na = 7.8 and 6.9, He = 0.71 and 0.68), while low genetic differentiation between populations (FST = 0.025) was observed. The newly characterised SSR markers extend the set of genetic markers available for F. excelsior and six other ash species for future studies on the genetic diversity and structure of ash populations.
摘要白蜡树(Fraxinus excelsior L.)是一种经济和生态上重要的树种,在欧洲受到白蜡树枯梢病的严重威胁。本研究基于已发表的引物,构建了一套新的单序列重复(Simple Sequence Repeat)标记。对德国两个白蜡树居群的40个SSR标记进行了扩增和多态性分析。此外,还评估了多态标记在6个亲缘种曲霉(f.a angustifolia、f.a ornus、f.a quadrangulata、f.t pennsylvania、f.a americana和f.t biltmoreana)间的可转移性。18个标记主要来自白蜡树枯梢病候选基因的基因区,其中3个标记与白蜡树枯梢病候选基因密切相关,其中10 ~ 17个标记可转移到6个亲缘种之一。两个白蜡树居群遗传多样性较高(Na = 7.8和6.9,He = 0.71和0.68),居群间遗传分化较低(FST = 0.025)。新鉴定的SSR标记为灰树和其他6种灰树的遗传标记集提供了进一步研究灰树种群遗传多样性和结构的基础。
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引用次数: 0
Traumatic ducts size varies genetically and is positively associated to resin yield of Pinus oocarpa open-pollinated progenies 创伤导管的大小在遗传上存在差异,且与开放授粉子代的树脂产量呈正相关
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0002
Irenka Fabián-Plesníková, C. Sáenz-Romero, T. Terrazas, Alejandro Reyes-Ramos, M. Martínez-Trujillo, J. Cruz-de-León, N. Sánchez-Vargas
Abstract Mexico ranks 5th in worldwide resin production. Pinus oocarpa is the most widely tapped pine tree in Mexico. Michoacán (central-western Mexico) is the first nationwide state producer of resin. Despite the P. oocarpa relevance, there is no genetic improvement program in the country for resin production. We evaluated the degree of genetic control for growth, anatomical traits, resin yield, and the correlation among them at an early age (five-years-old for growth, six for resin, and anatomical traits) in a P. oocarpa half-sib progeny trial. Families were originated from selected mother trees, based on their resin yield. We found significant genetic variation among families for stem volume (h2i= 0.12, h2f= 0.35), traumatic ducts (diameter: h2i= 0.63, h2f= 0.51; area: h2f= 0.81), and resin yield (individual and family narrow-sense heritability: h2i= 0.20, h2f= 0.52), and a positive correlation between diameter and area of traumatic ducts with resin yield (Pearson correlation: r= 0.73, p= 0.04; r= 0.71, p= 0.0497; respectively). Results suggest that the early selection (six-years-old) of superior P. oocarpa families, based on resin yield (estimated by microchipping technique), and/or based on larger diameter and area of traumatic ducts, appears to be a feasible strategy to develop seed orchards able to provide genetically improved seeds for intensive resin tree plantations. That would be an important alternative for a state as Michoacán, Mexico, where natural P. oocarpa stands are being replaced for avocado orchards for exportation.
墨西哥在全球树脂产量中排名第五。乌卡帕松是墨西哥种植最广泛的松树。Michoacán(墨西哥中西部)是全国第一家树脂生产商。尽管与P. oocarpa相关,但在国内没有树脂生产的遗传改良计划。我们在一项oocarpa半同胞后代试验中评估了生长、解剖性状、树脂产量的遗传控制程度,以及它们在早期(生长5岁,树脂和解剖性状6岁)的相关性。根据树脂产量,这些家族来自于选定的母树。我们发现,在茎体积(h2i= 0.12, h2f= 0.35)、创伤导管(h2i= 0.63, h2f= 0.51;面积:h2f= 0.81),树脂产率(个体和家族狭义遗传力:h2i= 0.20, h2f= 0.52),创伤导管直径和面积与树脂产率呈正相关(Pearson相关性:r= 0.73, p= 0.04;R = 0.71, p= 0.0497;分别)。结果表明,基于树脂产量(通过微芯片技术估计)和/或基于更大直径和面积的创伤导管,早期选择(6年生)优越的树科,似乎是开发能够为集约化树脂树种植园提供遗传改良种子的种子园的可行策略。对于墨西哥Michoacán这样的国家来说,这将是一个重要的选择,在那里,天然的牛油果林正被用于出口的牛油果果园所取代。
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引用次数: 2
Estimating of Additive, Dominance, and Epistatic Genetic Variance in Eucalypt Hybrid Population 桉树杂交群体加性、显性和上位遗传变异的估计
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0005
Makouanzi Ekomono Chrissy Garel, Vigneron Philippe
Abstract Additive, dominance and epistasis genetic variances were estimated from analysis of a clonally replicated full-sib progeny test grown in the Republic of Congo. Phenotypic variance components were estimated for ages 4 through 25 months for growth and at ages 8 and 18 months for ecophysiological traits. The estimation of genetics effects was derived from the individual mixed model. Genetic structure was incorporated into variances and covariance’s effects based on markers information. The detected genetic effects of epistasis are significant in some traits. This study shows that epistasis variance can be non-zero and contribute significantly to the genetic variability of growth and ecophysiological traits. We conclude that the epistatic effect for quantitative traits may exist, but estimates may not be obtained, either because the models used are inappropriate or because the epistasis variance is too small relative to other components of the genetic variance to be estimated.
通过对在刚果共和国生长的克隆复制全同胞后代试验的分析,估计了加性、显性和上位性遗传变异。对4 - 25月龄的生长和8 - 18月龄的生态生理性状的表型方差进行了估计。遗传效应的估计是由个体混合模型得出的。基于标记信息,将遗传结构纳入方差和协方差效应。在某些性状中检测到的上位遗传效应是显著的。本研究表明,上位性变异可以不为零,对生长和生理生态性状的遗传变异有重要影响。我们的结论是,数量性状的上位性效应可能存在,但可能无法得到估计,要么是因为使用的模型不合适,要么是因为上位性方差相对于遗传方差的其他组成部分太小而无法估计。
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引用次数: 0
In vitro amphidiploid induction of a distant hybrid Populus simonii × P. euphratica cv. ‘Xiaohuyang-2’ and its effect on plant morphology and anatomy
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0013
Xiao-Xiao Zhang, Ying Zhang, Xiaohong Cui, Dai-li Li, Hengyu Zhang, Wula Duan, Jun Wang
Abstract Highly gametic sterility of a distant hybrid Populus simonii × P. euphratica cv. ‘Xiaohuyang-2’ restricts its utilization in breeding programs of Populus. Amphidiploid induction by somatic chromosome doubling is expected to restore its gametic fertility. In this study, nodal-segment and leaf explants of ‘Xiaohuyang-2’ were used to induce chromosome doubling with colchicine in vitro. Although chromosome doubling of the nodal-segment explants only produced mixoploids, the treatments of leaf explants on adventitious bud regeneration medium successfully produced 4 amphidiploids, which might be attributed to the direct organogenesis of the adventitious buds on the leaf explants. This is the first report of amphidiploid induction in a distant hybrid between Populus section Tacamahaca and sect. Turanga. The highest amphidiploid induction frequency was 16.7 %. Both the explant survival rate and polyploidization frequency were significantly affected by colchicine concentration and exposure time. The amphidiploid plants significantly differed from the diploid and mixoploid plants in morphological and anatomical characteristics. They had larger, thicker, and greener leaves than the diploids and mixoploids. The increase in ploidy level also resulted in changes in stomatal features. The induced amphidiploid plants of the distant hybrid ‘Xiaohuyang-2’ are expected to play important roles in breeding programs of Populus in the future, which can be used as a bridge parent with the ability of unreduced gamete formation to cross with fast-growth germplasms to produce triploids pyramiding desirable traits of fast growth, easy cutting propagation, and salt and drought tolerances.
摘要远缘杂交胡杨(Populus simonii × P. euphratica cv)的高度配子不育性。“小虎阳2号”限制了其在杨树育种中的应用。体细胞加倍诱导双二倍体有望恢复其配子育性。本研究以‘小虎阳2号’的节段外植体和叶片外植体为材料,利用秋水仙碱在体外诱导染色体加倍。虽然节段外植体的染色体加倍只产生混合二倍体,但叶片外植体在不定芽再生培养基上的处理成功地产生了4个两二倍体,这可能是由于不定芽在叶片外植体上的直接器官发生。本文首次报道了塔卡玛哈卡杨与图兰加杨远缘杂交的双二倍体诱导。双二倍体的最高诱导率为16.7%。秋水仙碱浓度和暴露时间对外植体成活率和多倍体频率均有显著影响。两二倍体植物在形态和解剖特征上与二倍体和混合倍体植物有显著差异。它们的叶片比二倍体和混倍体更大、更厚、更绿。倍性水平的增加也导致气孔特征的变化。远缘杂交‘小虎阳2号’的双二倍体诱导植株在今后的杨树育种中具有重要的应用价值,可以作为具有未还原配子形成能力的桥梁亲本,与速生种质杂交,获得生长快、易扦插繁殖、耐盐耐旱等优良性状的三倍体。
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引用次数: 0
Growth performance of wild cherry (Prunus avium L.) clones in planted forests under different managements in Galicia, NW Spain 西班牙西北部加利西亚不同管理方式下人工林野生樱桃无性系的生长性能
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2022-01-01 DOI: 10.2478/sg-2022-0010
J. Fernández-Moya, I. Urbán-Martínez
Abstract Wild cherry (Prunus avium L.) is one of the main valuable temperate broadleaved species from Europe considering the market appreciation of their timber. In the present study we analyzed the growth performance of different wild cherry clones under different management treatments. A field trial was settled in Bosques Naturales S.A. “Sendelle” forests (Arzúa, Galicia, NW Spain) with a multifactorial design to analyze the effect of clones (4 clones), pruning intensity (33 % and 50 % of total height) and fertilization (3 treatments). It was monitored for 8 years. Clone was found out to be the most important variable in this study while tree growth was neither significantly affected by the pruning intensity nor the fertilization treatments. Clone selection is highlighted as a key issue in wild cherry commercial planted forests for timber production and the selected C-15 clone shows an excellent field performance compared to other clones based on the trial and the literature, reaching a DBH of 14 cm after eight years.
摘要野生樱桃(Prunus avium L.)是欧洲温带阔叶树种中最具价值的树种之一。本研究分析了不同管理处理下不同野生樱桃无性系的生长性能。在Bosques Naturales S.A.“Sendelle”森林(Arzúa, Galicia, NW Spain)进行了田间试验,采用多因子设计分析了无性系(4个无性系)、修剪强度(占总高度的33%和50%)和施肥(3个处理)的效果。它被监测了8年。在本研究中,无性系是最重要的变量,而修剪强度和施肥处理对树木生长均无显著影响。无性系选择是野生樱桃商业人工林木材生产的关键问题,根据试验和文献,选择的C-15无性系与其他无性系相比,表现出优异的田间表现,8年后达到了14厘米的胸径。
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引用次数: 0
Expression profiles of 11 candidate genes involved in drought tolerance of pedunculate oak (Quercus robur L.). Possibilities for genetic monitoring of the species. 有花序栎(Quercus robur L.)耐旱相关11个候选基因的表达谱对物种进行遗传监测的可能性。
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2021-01-01 DOI: 10.2478/sg-2021-0020
B. Trudić, Gordon Draškić, G. Le Provost, S. Stojnić, A. Pilipović, A. Ivezić
Abstract Pedunculate oak (Quercus robur L.) is one of the most significant broadleaved tree species in Europe. However, various abiotic and biotic agents have influenced pedunculate oak forests, among which drought stress has been frequently described as the main driver of this species forests decline. In this study we assessed relative expression profile of 11 candidate genes involved in many different metabolic pathways and potentially responsible for oak drought tolerance. The obtained results succeed in partially tackling drought tolerance mechanisms of targeted natural pedunculated oak population. This gene pool may represent a base for adaptation and therefore genetic diversity should be conserved. In this paper we described different expression responses of four pedunculate oak ecological groups, characterized by different physiological status (senescent vs vital) and flowering period (early (var. praecox) vs late (var. tardissima)). The most significant differences in relative gene expression levels are shown between the flowering period (tardissima (8 genes upregulated) vs praecox (3 genes upregulated)), more than a physiological status (sene-scent vs vital). Only three genes wrky53, rd22 and sag21 showed upregulated expression pattern in senescent physiological groups, indicating their possible role in the coping mechanisms of oak in stressed environment. Results showed interesting connections of relative gene expression values of identified drought-tolerance related genes with flowering period and provide further recommendations for adequate conservation and monitoring of this important oak gene pool in its southeast refugium.
有柄栎(Quercus robur L.)是欧洲最重要的阔叶树种之一。然而,各种非生物和生物因素影响着有花序栎林,其中干旱胁迫经常被描述为该物种森林衰退的主要驱动因素。在这项研究中,我们评估了11个候选基因的相对表达谱,这些基因参与了许多不同的代谢途径,并可能与橡树的抗旱性有关。所获得的结果成功地部分解决了目标自然有柄橡树群体的抗旱机制。这个基因库可能是适应的基础,因此应该保护遗传多样性。研究了不同生理状态(衰老vs生机)和花期(早(早熟)vs晚(姗姗来迟)的4个有花序栎生态类群的不同表达响应。相对基因表达水平最显著的差异表现在花期之间(迟蒿(8个基因上调)与先梅(3个基因上调)),而不是生理状态(衰老与重要)。只有wrky53、rd22和sag21三个基因在衰老生理组中表达上调,提示它们可能参与橡树逆境应对机制。结果表明,所鉴定的抗旱性相关基因的相对表达值与开花期之间存在着有趣的联系,为进一步保护和监测东南地区这一重要的橡树基因库提供了建议。
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引用次数: 1
Genetic tolerance to Ceratocystis wilt in melina (Gmelina arborea Roxb.) melina (Gmelina arborea Roxb.)对枯萎病的遗传耐受性
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2021-01-01 DOI: 10.2478/sg-2021-0017
Dawa Méndez-Álvarez, Olman Murillo-Gamboa, Yorleny Badilla-Valverde, William Hernández-Castro
Abstract Gmelina arborea is the second most planted tree species in Costa Rica, and one of the most important in several Latin American countries. In recent years, a disease caused by Ceratocystis fimbriata, has spread throughout Latin America, leading to the complete root of the tree in a few months. This study was conducted to evaluate the incidence, severity and genetic tolerance control of the Ceratocystis wilt in G. arborea. Data is based on a 2.4-year-old clonal trial, with 34 elite genotypes, at Río Jiménez, Guácimo in the Costa Rican Caribbean region. The variables diameter at breast height (DBH), total height, disease incidence and disease severity were evaluated. A pathogenicity index was proposed as a new selection criterion. SELEGEN (EMBRAPA) software was used for the evaluation of the genetic control in all traits investigated. Overall incidence rate was 39 % and severity was 13 %. Mean clonal heritability for severity was h2mc= 0.59 and for incidence h2mc= 0.47, these values were high evidencing that both traits are under a strong genetic control. The pathogenicity index becomes a useful discrimination criterion for ranking genotypes by their disease tolerance. An infection pattern inversely proportional to the DBH was observed. Although none of the 34 genotypes evaluated recorded total resistance to C. fimbriata, eight of these genotypes show strong genetic tolerance to the disease, and high productivity. Their use can be encouraging and will contribute to the reduction of Ceratocystis wilt impact in the country, as well as in the tropical Latin America region.
林荫小木桐(Gmelina arborea)是哥斯达黎加种植面积第二大的树种,也是几个拉丁美洲国家最重要的树种之一。近年来,一种由毛状角鼻虫引起的疾病在整个拉丁美洲蔓延,导致树木在几个月内完全扎根。本研究对青木角鼻虫的发病、严重程度及遗传耐受性控制进行了评价。数据基于哥斯达黎加加勒比地区Río jimsamnez, Guácimo进行的一项持续2.4年的克隆试验,其中包含34个优秀基因型。评估乳房高度直径(DBH)、总高度、疾病发生率和疾病严重程度等变量。提出了致病性指数作为新的选择标准。采用SELEGEN (EMBRAPA)软件对所有性状的遗传控制进行评价。总发病率为39%,严重程度为13%。严重程度的平均克隆遗传率为h2mc= 0.59,发生率的平均克隆遗传率为h2mc= 0.47,表明这两种性状均受很强的遗传控制。致病性指数是根据基因型的耐受性对其进行排序的一个有用的判别标准。感染模式与胸径成反比。虽然34个基因型中没有一个记录到对红僵菌的完全抗性,但其中8个基因型对该疾病表现出很强的遗传耐受性和高产量。它们的使用是令人鼓舞的,并将有助于减少角鼻虫对该国以及热带拉丁美洲区域的影响。
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引用次数: 1
Obituary - Dr. Hans J. Muhs (1939-2021) Hans J. Muhs博士(1939-2021)
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2021-01-01 DOI: 10.2478/sg-2021-0015
B. Degen, M. Fladung
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引用次数: 0
The traceability of Eucalyptus clones using molecular markers 利用分子标记研究桉树无性系的可追溯性
IF 1 4区 农林科学 Q3 FORESTRY Pub Date : 2021-01-01 DOI: 10.2478/sg-2021-0019
D. Torres-Dini, Leonardo Delgado-Cerrone, L. Luna, F. Resquin, A. Aguiar, A. Sebbenn
Abstract The improvement of Eucalyptus clones plays a crucial role in modern silviculture. This study used a set of 17 microsatellite loci to analyze the genetic diversity and structure of 107 elite clones (80 E. grandis and 27 E. globulus). All clones were cultivated in Uruguay and were sourced from three different providers. Using the fingerprinting technique, an exclusive molecular profile was assigned for each clone, and the genotyping reaction showed differences between the two species. The cumulative probability of identifying two random individuals that share the same genotype (PI) with all 17 loci, was estimated as low for E. grandis (1.18×10-15) and E. globulus (4.03×10-14). The combined PIsibs was (1.05×10-5) and (2.17×10-5) for E. grandis and E. globulus, respectively. A total of 180 alleles were detected for E. grandis and 100 for E. globulus. We found a high mean number of alleles per locus (10 for E. grandis and 6 for E. globulus), and the results for mean polymorphic information content (PIC ) were (0.648) and (0.548), respectively. The observed heterozygosity (Ho) ranged from 0.216 to 0.838 (mean = 0.509) for E. grandis and 0 to 1 (mean = 0.566) for E. globulus. Two core sets of seven EST-SSR loci were identified for each species. These markers revealed unambiguous fragment amplification, providing a minimum number of SSRs for effective clonal identification. The genetic structure analysis suggests that the germplasm of the E. grandis population is structured in four clusters, while the E. globulus population consists of two clusters.
摘要桉树无性系的改良在现代造林中起着至关重要的作用。本研究利用17个微卫星位点,对107个优质无性系(80个大田鸡和27个球田鸡)的遗传多样性和结构进行了分析。所有无性系均在乌拉圭栽培,来源为三个不同的供应商。利用指纹图谱技术,为每个克隆分配了一个专属的分子图谱,基因分型反应显示了两个物种之间的差异。据估计,在大e.g grandis (1.18×10-15)和globulus (4.03×10-14)中,发现两个随机个体具有相同基因型(PI)的累积概率很低。大e.g grandis和globulus的联合pisib分别为(1.05×10-5)和(2.17×10-5)。大大肠杆菌共检测到180个等位基因,球状大肠杆菌共检测到100个。结果表明,每个位点的平均等位基因数较高(大叶芝为10个,球叶芝为6个),平均多态性信息含量(PIC)分别为0.648和0.548。观察到的杂合度(Ho)范围为0.216 ~ 0.838(平均= 0.509),大棘球棘球绦虫的杂合度(Ho)为0 ~ 1(平均= 0.566)。每个物种共鉴定出2组7个EST-SSR位点。这些标记显示了明确的片段扩增,为有效的克隆鉴定提供了最小数量的SSRs。遗传结构分析表明,大榄居群的种质结构为4个簇,而球榄居群的种质结构为2个簇。
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引用次数: 0
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Silvae Genetica
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