Single nucleotide polymorphisms of the KIT and KITLG genes in pigs

Here, we report the variability in the KIT tyrosine kinase receptor and its ligand KITLG genes by determining single nucleotide polymorphisms (SNPs) in 384 individuals including 11 pig breeds, two synthetic-line cross pigs, two cross breeds, and one Japanese wild boar. SNPs and indels within the coding sequence regions of KITLG and KIT and their 5'-flanking regions were detected by aligning sequences from eight pigs, and subsequently the SNPs were genotyped using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF-MS). Principal component analysis using allele frequencies in the SNP locus showed a distant relationship between Asian and Euro-American pig groups, except for Berkshire and Tokyo X breeds. These breeds were located within the mid-portion of the distribution in the first principal component. The Hampshire breed was distant from the other pig groups on the axis of the second principal component. Haplotype frequencies that were deduced using non-synonymous substitutions of the KIT gene revealed the uniqueness of Landrace, Large White, Middle White, and three-way cross pigs (LWD) and of the Hampshire breed. On the other hand, the haplotypes of KITLG and KIT detected in the Berkshire breed were prevalent in Asian pig groups. This tendency is different from that observed in other Euro-American pig breeds.     

阿坝藏族羌族自治州若尔盖地区藏猪mtDNA D-Loop的遗传多样性分析

旨在通过获得和对比若尔盖地区藏猪mtDNA D-Loop高变区的部分序列,为该地区藏猪遗传资源的保护与利用提供参考。本试验收集了若尔盖地区9个乡镇共80头藏猪耳组织,以甘南州藏猪mtDNA D-Loop基因序列为模板设计引物,采用PCR扩增测序技术获得了80条核苷酸序列,并采用生物信息学的数据处理方法进行分析。结果表明,若尔盖地区藏猪mtDNA D-Loop高变区（435 bp）的A+T含量（56.46%）明显高于G+C含量（43.54%）,存在碱基偏倚性现象;在80条长度为435 bp的序列中,共检测到14个变异位点,鉴定了17个单倍型,单倍型多样度（Hd）、核苷酸多样度（Pi）和平均核苷酸差异数（k）分别为0.881、0.004 66和2.028,其中Hd、Pi和k在降扎乡藏猪群体中最高,Pi和k在占哇乡藏猪中最低;共享单倍型8个,特有单倍型9个,且若尔盖地区不同藏猪群体间特有单倍型数差异较大,其中,降扎乡藏猪的特有单倍型数量最多,占单倍型总数的17.65%（3/17）;Hap_1和Hap_15单倍型是4个乡镇（降扎乡、益哇乡、热尔乡和冻列乡）群体的共享单倍型,表明这4个乡镇藏猪群体存在两个共同的母系祖先单倍型。若尔盖地区藏猪的平均遗传距离为0.004 66,其中降扎乡和冻列乡藏猪间的遗传距离最大为0.006 90,包座乡和益哇乡藏猪间的遗传距离最小为0.002 16;构建的NJ分子系统进化树将若尔盖地区藏猪分为2支,而加入中国地方家猪、野猪和引种猪后,若尔盖地区藏猪在进化树中比较分散,说明该地区藏猪母源血统遗传复杂,彼此之间基因交流多。本研究进一步证实了若尔盖地区藏猪比西藏林芝、山南、日喀则、甘孜州、阿坝州藏猪的遗传多样性程度高,受到人工选择强度低,应强化对若尔盖地区藏猪遗传资源的保护与利用。     

Variation in the prion protein sequence in Dutch goat breeds

Scrapie is a neurodegenerative disease occurring in goats and sheep. Several haplotypes of the prion protein increase resistance to scrapie infection and may be used in selective breeding to help eradicate scrapie. In this study, frequencies of the allelic variants of the PrP gene are determined for six goat breeds in the Netherlands. Overall frequencies in Dutch goats were determined from 768 brain tissue samples in 2005, 766 in 2008 and 300 in 2012, derived from random sampling for the national scrapie surveillance without knowledge of the breed. Breed specific frequencies were determined in the winter 2013/2014 by sampling 300 breeding animals from the main breeders of the different breeds. Detailed analysis of the scrapie‐resistant K₂₂₂ haplotype was carried out in 2014 for 220 Dutch Toggenburger goats and in 2015 for 942 goats from the Saanen derived White Goat breed. Nine haplotypes were identified in the Dutch breeds. Frequencies for non‐wild type haplotypes were generally low. Exception was the K₂₂₂ haplotype in the Dutch Toggenburger (29%) and the S₁₄₆ haplotype in the Nubian and Boer breeds (respectively 7 and 31%). The frequency of the K₂₂₂ haplotype in the Toggenburger was higher than for any other breed reported in literature, while for the White Goat breed it was with 3.1% similar to frequencies of other Saanen or Saanen derived breeds. Further evidence was found for the existence of two M₁₄₂ haplotypes, M₁₄₂/S₂₄₀ and M₁₄₂/P₂₄₀. Breeds vary in haplotype frequencies but frequencies of resistant genotypes are generally low and consequently selective breeding for scrapie resistance can only be slow but will benefit from animals identified in this study. The unexpectedly high frequency of the K222 haplotype in the Dutch Toggenburger underlines the need for conservation of rare breeds in order to conserve genetic diversity rare or absent in other breeds.     

Hidden Diversity in an Antarctic Algal Forest: Metabolomic Profiling Linked to Patterns of Genetic Diversification in the Antarctic Red Alga Plocamium sp.

The common Antarctic red alga Plocamium sp. is rich in halogenated monoterpenes with known anticancer and antimicrobial properties and extracts of Plocamium sp. have strong ecological activity in deterring feeding by sympatric herbivores. Plocamium sp. collected near Anvers Island, Antarctica showed a high degree of secondary metabolite diversity between separate individuals. GC/MS results revealed 15 different combinations of metabolites (chemogroups) across individuals, which were apparent at 50% or greater Bray–Curtis similarity and also clearly distinguishable by eye when comparing chromatographic profiles of the secondary metabolomes. Sequencing of the mitochondrial cox1 gene revealed six distinct haplotypes, of which the most common two had been previously reported (now referred to as Haplotypes 1 and 2). With the exception of one individual, three of the chemogroups were only produced by individuals in Haplotype 1. All the other 12 chemogroups were produced by individuals in Haplotype 2, with five of these chemogroups also present in one of the four new, less common haplotypes that only differed from Haplotype 2 by one base pair. The functional relevance of this metabolomic and genetic diversity is unknown, but they could have important ecological and evolutionary ramifications, thus potentially providing a foundation for differential selection.     

福建省部分马铃薯产区晚疫病菌群体结构研究

为明确福建省部分马铃薯产区晚疫病菌群体结构,2017年-2019年在龙海市、福安市、霞浦县共分离获得96株马铃薯晚疫病菌。采用对峙培养法、鉴别寄主法和PCR-RFLP法对这些菌株的交配型、生理小种及线粒体DNA (mtDNA)单倍型进行分析。交配型测定结果显示,除了福建省福安市有5株(5.21%)为A1交配型外,其余91株(94.79%)均为自育型,未发现A2交配型及A1A2型菌株。从96个菌株中检测出16个生理小种类型,龙海市和福安市的优势生理小种均是可克服11个显性抗病单基因的1.2.3.4.5.6.7.8.9.10.11,霞浦县的优势生理小种为1.2.3.4.5.6.7.8.10。供试菌株均至少含有6个毒性基因。mtDNA单倍型共检测出3种类型,其中55个菌株(龙海市22株、福安市8株、霞浦县25株)为Ⅰa型,占57.29%,32个菌株(龙海市2株、福安市25株、霞浦县5株)为Ⅱa型,占33.33%,9个菌株(福安市7株、霞浦县2株)为Ⅱb型,占9.37%。研究结果表明,福建省马铃薯晚疫病菌群体遗传多样性日趋复杂。     

Population studies on <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on potatoes and tomatoes in southern Germany

Fifty-seven isolates of Phytophthora infestans from blighted potato foliage were collected in 1995 in southern Germany and analysed for mating type and sensitivity to metalaxyl. Fifty-six of them were characterised as A1 and one as A2 mating types. Resistance to metalaxyl was observed frequently: 53 isolates were resistant, three were partially sensitive, and one was sensitive. In a subsequent field study in 1999, 84 isolates collected from blighted potato and tomato foliage were analysed for mating type. Seventy-two were characterised as A1 and twelve as A2 mating types. The response of 76 isolates to metalaxyl and to propamocarb was tested. The majority (42) of the 76 isolates was classified as resistant to metalaxyl; 31 were partially sensitive and only three isolates were sensitive. The results with propamocarb were less discrete; 10 isolates were classified as resistant and three were clearly sensitive. AFLP fingerprinting was used to examine the genetic structure of the southern German P. infestans population collected in 1999 and indicated that the tested population can be sub-divided into a tomato group, a potato group and a mixed group containing isolates collected from both crops. The presence of Ia and IIa mitochondrial DNA haplotypes indicates that the German P. infestans isolates belong to the new pathogen population that has also been reported in neighbouring regions of Europe. The present study indicates that at the beginning of the season only a few genotypes were present, and the population became genetically more variable at the end of the growing season.     

利用线粒体DNA标记分析中国东南沿海拟穴青蟹种群遗传结构

为研究我国东南沿海拟穴青蟹(Scylla paramamosain)的种群遗传结构,对10个地理种群130只拟穴青蟹的线粒体DNA（mitochondrial DNA,mtDNA）细胞色素氧化酶亚基I（COI）基因片段序列进行分析。522bp的DNA片段共发现17个变异位点,定义了21种单倍型,其中单倍型2为10个地方种群的共享单倍型,占个体总数的56.15％,而大部分单倍型为稀有单倍型,只在少数种群的个别个体中出现。10个种群的单倍型多样性水平为h=0.6738,核苷酸多样性水平为π=0.1987%,基本呈由南到北递减的趋势。10个种群的总体遗传分化程度较低（FST＝0.05左右）,但是极为显著（P<0.005）。基于单倍型频率和序列遗传距离法分析的共同结果,广西北海种群与大多数种群的遗传分化显著,而海南三亚种群分别与海南红树林和广东台山种群遗传分化显著。Mantel检验显示种群间的遗传分化程度与地理距离没有显著的相关性。分子进化中性检验结果表明自然选择在分子进化过程中起了重要作用,并暗示该物种在最近经历了一个快速的种群爆发及扩张事件。     

柑橘CCD基因家族鉴定及CcCCD4a对果肉颜色的影响

【目的】研究类胡萝卜素裂解双加氧酶（CCD）基因家族在柑橘基因组中的分布、结构及进化,对CcCCD4a在果肉颜色形成过程中的表达及其在不同果肉颜色的柑橘种质中的基因型进行研究,为开发用于果肉颜色的分子辅助育种标记奠定基础。【方法】根据已报道的CCD,采用同源比对法检索柑橘基因组中的CCD家族基因（CcCCD）。采用生物信息学软件构建系统进化树,进行亚细胞定位预测,预测蛋白质的相对分子质量与等电点（pI）等理化性质,预测保守motif,绘制家族基因Scaffold定位图。利用实时荧光定量PCR技术（qRT-PCR）分析CcCCD4a在柑橘果实颜色发育过程中的表达模式,利用测序技术鉴定30个柑橘品种的CcCCD4a基因型,采用Tassel软件进行单倍型分析。【结果】从克里曼丁橘（Citrus clementina）基因组中鉴定出14个CcCCD基因家族成员,可将其分为5个亚家族,即CcCCD1、CcCCD4、CcCCD7、CcCCD8和CcNCED。该家族蛋白理论等电点分布在6.05—8.53,编码氨基酸数目介于412—611个;亚细胞定位预测结果显示该基因家族成员主要位于叶绿体和细胞质中;聚类分析发现,CCD8亚家族与其他家族成员遗传距离较远,柑橘中各CCD均能在其他物种中找到同源基因;Scaffold定位分析发现,14个CCD家族成员成员分布在除5号Scafflod外的所有Scafflod上,且分布不均匀。对10个柑橘品种在4个时期的果肉色泽进行表型鉴定,随着果实趋于成熟,果肉的色调角（h）逐步下降,果肉颜色逐步加深;CcCCD4a在不同柑橘品种中相对表达量存在显著差异,果肉颜色为浓橙红色的品种CcCCD4a表达量显著低于果肉为橙色或浅橙黄色的品种(P<0.05),CcCCD4a相对表达量与色调角呈显著正相关(P<0.05);对30个柑橘品种进行测序分析,发现单倍型hap-1、hap-4和hap-5为果肉浓橙红色品种优势单倍型。【结论】‘克里曼丁’橘包含14个CCD基因家族成员,各成员均含有RPE65保守结构域,并定位于细胞的不同位置,分布在不同的Scaffold上。CcCCD4a参与柑橘果肉颜色的形成,其基因相对表达量与果肉色调角呈显著正相关,可作为潜在的柑橘果实颜色的辅助育种标记,尤其是单倍型hap-1、hap-4、hap-5与果肉红色的关联度较高,对颜色育种的早期杂种群体筛选有一定帮助。     

入侵我国中南三省(区)草地贪夜蛾的单倍体型和群体遗传结构分析

【目的】揭示入侵我国中南三省(区)的草地贪夜蛾Spodoptera frugiperda种群间可能存在的遗传变异,探讨其入侵来源、入侵路线和扩散方式,为制定有效控制和阻击草地贪夜蛾的方案提供理论依据和遗传信息。【方法】利用分子标记技术从线粒体DNA和核DNA角度研究草地贪夜蛾种群的遗传多样性和种群结构。对采集自我国广东、广西和湖南3个省(区)的草地贪夜蛾,利用COI和Tpi基因片段进行了单倍体型分析,利用9个微卫星标记分析种群间的遗传结构和遗传多样性。【结果】供试样品中,除了湖南张家界群体有2个样品为COI玉米型单倍体型、1个样品为杂合型,其他群体的个体均为COI水稻型单倍体型。基于SSR标记的群体遗传分析结果表明,被检测的群体遗传多样性较巴西等国家的美洲群体低,各群体遗传分化不明显。广东东莞群体与湖南张家界群体的遗传距离最近,与广西南宁群体的遗传距离较远。【结论】遗传距离与实际距离之间不一定呈正相关,通过遗传结构的分析有助于推测草地贪夜蛾扩散过程中,天气背景场、地理隔离和人为传播各自所起的作用。入侵湖南省张家界的草地贪夜蛾出现了低频率COI玉米型和高频率的CO I水稻型单倍体型,说明虫源的非单一性;草地贪夜蛾在入侵地定殖时间较短,目前未形成明显的遗传分化。     

Population structure and species delimitation of rice white tip nematode,Aphelenchoides besseyi (Nematoda: Aphelenchoididae), in China

Aphelenchoides besseyi is a major nematode pathogen in rice known as the causal agent of white tip disease. In the present study, A. besseyi was recovered from 32 seed samples collected from the major rice-growing regions in China. Nine mitochondrial COI haplotypes and 95 ITS genotypes were identified, suggesting a high genetic diversity and endemism level of A. besseyi in China. Therefore, A. besseyi is likely to have experienced a long history of host–parasite coevolution in China, rather than being recently introduced from other countries. The results of population structure analysis indicated the presence of four clusters within Chinese A. besseyi, but these did not correspond to their geographic distribution. Haplotype H2 was found to be the most widespread. Haplotypes H9 and H13 were most genetically divergent and have been recognized as two cryptic species by molecular species delimitation methods, and with limited support from morphometric measurements. The COI-based phylogeny suggested the ability to parasitize rice has independently evolved at least four times in the genus Aphelenchoides.