四倍体马铃薯块茎蛋白含量分子标记的开发与验证

块茎蛋白含量是马铃薯的重要品质性状之一, 相关分子标记的开发研究较少。本研究以马铃薯高蛋白品种大西洋、低蛋白品种定薯1号及包含173份子代的蛋白含量分离群体为材料, 通过高通量简化基因组测序技术与集群分离分析(BSA)相结合, 开发了SCAR8-107标记, 并以此标记对分离群体的74份高蛋白、42份低蛋白子代及54个四倍体马铃薯品种验证表明, SCAR8-107标记在后代分离群体和四倍体马铃薯品种中的检测结果与表型蛋白含量的对应度分别达到了90.51%和72.97%, 经过Pearson’s双侧相关性分析, 相关性均达到了极显著水平。SCAR8-107的开发对于四倍体马铃薯块茎蛋白含量标记辅助选择与加速马铃薯品质育种具有重要意义。     

基于2b-RAD测序的四倍体马铃薯熟性相关的分子标记开发

马铃薯熟性是由多基因控制的数量性状,是我国不同栽培区划选择适宜品种的重要指标之一。本研究以晚熟品种中薯18号和早熟品种中薯5号及其F₁分离群体为材料, 2018—2019年连续2年对“中薯18号(母本)×中薯5号(父本)”杂交分离群体进行熟性评价,从中筛选出极端晚熟和极端早熟的基因型各30个,并分别构建极端晚熟和极端早熟基因组DNA混池。利用简化基因组2b-RAD (2b-restriction site-associated DNA)技术测序,寻找差异标签开发出3个与熟性连锁的分子标记SCARA2-2、SCARA4-21和SCARA5-16,3个分子标记联合使用对熟性分离群体子代进行验证,晚、早熟表型符合率分别到达了87.5%和93.0%,这些分子标记的开发和联合使用对辅助马铃薯熟性选择具有重要的参考价值。     

马铃薯块茎形状CAPS标记的开发与验证

马铃薯块茎形状(薯形)是选育马铃薯品种尤其是加工品种最重要的性状之一。前人研究表明薯形是由位于第10染色体的单基因Ro控制的,并且圆形对长形为显性。本研究以二倍体马铃薯长薯形基因型10618-01和圆薯形基因型320-02及其213个F₁代薯形分离群体为材料,基于马铃薯基因组相关序列信息结合分离群体分组分析法开发标记,获得了一个CAPS标记1137-CAPSVI。利用该标记对53份不同薯形的四倍体马铃薯高代品系进行验证,结果表明该标记的选择准确率达83.02%,进一步的Pearson双侧相关分析显示其相关性达极显著水平,且该标记对圆薯形材料的选择准确率更高,达91.42%。该CAPS标记的获得对于四倍体马铃薯分子标记辅助育种,加速马铃薯品种尤其是加工专用型品种的选育具有重要意义。     

马铃薯块茎形状基因CAPS标记的开发与验证

马铃薯块茎形状(薯形)是选育马铃薯品种尤其是加工品种最重要的性状之一。前人研究表明薯形是由位于第10染色体的单基因Ro控制的,并且圆形对长形为显性。本研究以二倍体马铃薯长薯形基因型10618-01和圆薯形基因型320-02及其213个F1代薯形分离群体为材料,基于马铃薯基因组相关序列信息结合分离群体分组分析法开发标记,获得了一个CAPS标记1137-CAPSVI。利用该标记对53份不同薯形的四倍体马铃薯高代品系进行验证,结果表明该标记的选择准确率达83.02%,进一步的Pearson双侧相关分析显示其相关性达极显著水平,且该标记对圆薯形材料的选择准确率更高,达91.42%。该CAPS标记的获得对于四倍体马铃薯分子标记辅助育种,加速马铃薯品种尤其是加工专用型品种的选育具有重要意义。     

马铃薯品种中薯3号杂交后代熟性分析

早熟马铃薯品种熟期短,经济效益高,已经成为农民增收致富的重要作物之一。为了解马铃薯熟性的遗传规律,为早熟品种选育的亲本选配提供依据,以早熟品种中薯3号为亲本分别与早熟、中早熟和晚熟亲本品种杂交形成的6个杂交组合后代为材料,对不同类型的杂交组合后代的熟性和产量相关性状进行了分析,结果表明:熟性是由多基因控制的、存在细胞质效应的数量性状;配制早熟×晚熟组合既能保证杂交后代中有较高比例的早熟和中早熟材料,也能保证杂交后代中有较高比例的高产材料;中薯3号是一个优良的、配制早熟杂交组合的育种材料。     

水稻显性早熟基因Ehd的SSR标记定位

以籼稻品种广陆矮4和粳稻品种台中65为亲本构建高世代回交分离群体,选用分布于水稻全基因组的145个SSR标记对亲本及抽穗期早熟基因进行分析.结果表明,114个标记在亲本间具有多态性,多态率78.6%;在BC3F1群体中,检测到10个标记的基因型来源于供体亲本广陆矮4号;在BC3F2定位群体中,早熟植株数与晚熟植株数的分离比例为3:1,早熟植株平均比晚熟植株提早抽穗21 d;通过SSR标记与抽穗期共分离分析将显性早熟基因Ehd界定在分子标记RM271和RM258之间;Ehd与标记RM184和RM271紧密连锁,遗传距离分别为2.6 cM和2.1 cM,此结果为该基因分子标记辅助选择奠定了基础.     

青海省马铃薯主要栽培品种的SSR遗传多样性

利用SSR标记分析了12份马铃薯栽培品种的遗传多样性及其亲缘关系,结果表明:(1)30对SSR引物共扩增出388条带,其中多态性条带为359条.(2)12份材料的遗传距离介于0.238 5～0.4801 cM之间,平均值为0.397 6 cM.(3)利用由非加权类平均法聚类分析表明,在遗传相似系数0.63处将12份材料被划分为五类:两类为晚熟(其中从国际马铃薯中心引进的青薯9号品种为一类;青薯168,2号,3号,4号品种为另一类),两类为早熟(其中青薯5号品种为一类;青薯8号品种和02-1-1品系为另一类),一类为中熟(因育成的地域不同而分为两个亚类).结果表明,青海省现有主要栽培的马铃薯品种遗传多样性水平较低.为拓宽育成品种的遗传基础,应充分发掘野生二倍体的遗传潜力.     

马铃薯淀粉含量相关SSR分子标记的开发及候选基因的初步确定

淀粉含量是衡量马铃薯品质的重要性状之一,本试验以高淀粉四倍体马铃薯野生种‘YSP-4’与低淀粉四倍体马铃薯品系‘MIN-021’杂交获得的F2代分离群体为材料,基于BSA分析技术,开发了2个与马铃薯淀粉含量紧密连锁的SSR分子标记SSR-152和SSR-174,并对其进行单标记分析验证。本研究对这2个标记片段进行了回收、纯化、测序及blastn序列比对发现,与基因XM＿006348370.1的同源性分别为96.63%和98.04%,该基因可能与马铃薯淀粉合成相关基因Patatin和耐旱基因有关,通过调控糖的合成与代谢,进而影响淀粉含量的积累,初步确定为候选基因。这2个标记为马铃薯淀粉相关候选基因的筛选、克隆以及开展分子标记辅助育种等研究奠定了基础。     

RYSC3和Rxsp分子标记在马铃薯抗病育种中的应用

马铃薯Y病毒(PVY)和马铃薯X病毒(PVX)是对马铃薯的产量和品质影响最主要的两种病毒,马铃薯中含有对PVY和PVX具有极端抗性的基因Ryadg和Rx,聚合抗性基因是防治PVY、PVX对马铃薯生产影响最有效的策略。本研究利用与Ryadg和Rx紧密相连的分子标记RYSC3和Rxsp,对青海省主栽马铃薯品种青薯9号与大西洋品种及其杂交F1代进行了标记检测。结果显示,亲本青薯9号仅含RYSC3标记,亲本大西洋仅含Rxsp标记;在198个基因型的F1代分离群体中,含RYSC3标记的材料有91份,含Rxsp标记的材料有86份,同时含Rxsp和RYSC3标记的材料共41份;RYSC3和Rxsp标记基因型的分离比例在5%显著水平下经卡平方检验都符合孟德尔分离比例1:1,抗病亲本均属于单显性基因型,基因型符合Rrrr×rrrr。本研究以期为青海省马铃薯抗病毒新品种的选育和早期鉴定提供科学依据。     

290个吉林省审定玉米品种SSR-DNA指纹构建及遗传多样性分析

吉林省审定玉米品种标准指纹构建及遗传多样性分析,可为其品种审定、保护、市场监测等提供核心数据支撑,同时对于品种选育、种质创新也具有重要意义。本研究利用40对SSR标记构建了吉林省2010—2017年审定的290个玉米品种标准SSR-DNA指纹并进行了遗传多样性分析。结果表明,40对SSR标记检测到等位基因数平均值为11.85个, PIC平均值为0.64;品种杂合基因型频率平均值为0.66;品种间差异位点数目均≥2个。6个熟期组的遗传多样性分析显示,中晚熟和中熟审定的品种较多占71.72%,极早熟、早熟、晚熟审定品种较少;早熟组具有基因多样性参数最高值0.69,品种数量最多的中晚熟组具有相对低的基因多样性值。聚类结果显示,吉林省审定推广种植品种被划分为3类,分别是甜糯爆裂、极早熟和早熟类、偏中或晚熟类;偏中或晚熟的品种显示了两大趋势,代表了选用杂优群的差异、品种选育单位的不同、品种类型的差异等。本研究为吉林省玉米品种管理、品种选育、种质创新提供了重要的理论和数据基础。     

19份马铃薯品种的SSR分析

为了解19份马铃薯品种资源的遗传差异及亲缘关系,利用筛选出的10对SSR适宜引物对19个供试材料的基因组DNA进行PCR扩增,得到稳定、清晰的SSR条带163个,其中多态性条带143个,其多态性比率占87.73%。19份马铃薯材料间的遗传距离(GD)介于0.2415~0.7048之间,平均GD值为0.4090。以GD值0.66为基准,将19份马铃薯品种分为5类:青薯9号、冀张薯14和冀张薯8号为一类,大百花(2191)、冀张薯5号、中薯19号、雪川1310和华颂11为一类,冀张薯12号、中薯22号和华颂7号为一类,红玫瑰、希森8号、5 p-40和希森6号为一类,中薯10号、中薯18号、华颂34和华颂33为一类。     

SCAR and RAPD markers associated with 18-carbon fatty acids in rapeseed, Brassica napus

Breeding rapeseed for enhanced oil quality includes the development of varieties with low linolenic acid content. The breeder also aims to develop varieties with a high linoleic acid content because of its nutritional value. Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers have been developed for linolenic acid content, but they are not best suited for a direct application in marker-assisted selection. The RFLP technique is too complex and time-consuming and RAPD markers lack codominance, precluding the distinction of homozygous from heterozygous individuals. In this report the conversion of a RAPD marker to a codominant sequence characterized amplified region (SCAR) marker named L1L9 is described. One of the alleles consisting of an 899 bp fragment (allele A), is associated with low linolenic acid content. The other allele consists of an 641 bp fragment (allele B) and is associated with high linolenic acid content. This marker explains approximately 25% of the genetic variation for this trait. Linkage analysis in the mapping population indicates that the SCAR marker probably tags an ω-3 desaturase gene in B. napus. Two RAPD markers were found to be associated with oleic/linoleic acid content. Markers M14-350 and I06-650 explained approximately 10% and 7% of the genetic variation for linoleic acid content, respectively. These two markers were found linked at 12.3cM in the segregating B. napus F₂ progeny used for mapping. All the markers reported in this paper should be useful in breeding programmes for developing high linoleic and low linolenic acid rapeseed varieties.     

基于侯选基因标记的四倍体马铃薯休眠QTL关联分析

休眠期是马铃薯(SolanumtuberosumL.)重要的块茎性状之一,寻找调控马铃薯块茎休眠的关键基因,揭示其分子机制以选育具有适宜休眠期长度的马铃薯品种,对于解决当前马铃薯产业中过长或过短休眠期带来的经济损失和食品安全隐患等问题十分关键。前期研究在二倍体马铃薯连锁群体中定位了6个加性休眠QTL,本研究拟在四倍体马铃薯育种材料中验证这些休眠QTL。基于休眠QTL连锁的候选基因标记,采用混合线性模型(MLM),模型中考虑群体结构和亲缘关系(Q+K),在四倍体马铃薯自然群体St-hzau中对马铃薯块茎休眠期进行了关联分析。5号染色体上休眠QTL DorB5.3连锁的候选基因标记S199₃00和GWD (根据葡聚糖水双激酶α-glucan water dikinase基因设计)与马铃薯块茎休眠期具有显著的关联(P<0.05),分别解释了休眠期表型变异的7.8%和3.2%,分别能增加休眠期7.1 d和4.5 d,即在二倍体马铃薯连锁群体中定位的稳定主效休眠QTL DorB5.3在四倍体马铃薯关联群体St-hzau中也表现显著, DorB5.3的稳定性在关联分析结...     

Population structure and linkage disequilibrium in diploid and tetraploid potato revealed by genome‐wide high‐density genotyping using the SolCAP SNP array

Genome‐wide association (GWA) mapping in potato requires high‐density genotyping. With the Illumina SolCAP potato single‐nucleotide polymorphism (SNP) array, a first tool for GWA mapping in potato became available. Thirty‐six tetraploid varieties and eight diploid breeding clones were genotyped for 8303 SNPs using this array. The objectives of our study were to examine in this set of germplasm: (i) the degree of polymorphism of the SolCAP SNPs in European germplasm, (ii) the population structure, (iii) temporal trends of genetic diversity and (iv) the genome‐wide extent of linkage disequilibrium (LD). Three‐quarters of the SNPs were polymorphic. In the principal coordinate analysis, a clear separation of tetraploid from diploid genotypes was observed, whereas no distinct subgroups among the tetraploid varieties were detected. The nonlinear trendline of the LD measure vs. the physical map distance decayed within 275 bp to an value of 0.10, indicating that theoretically, about 3 million equally distributed SNPs are required for GWA mapping in this diverse set of germplasm. As the LD decay changes with the population selected for GWA mapping, the number of required markers might be different in other germplasm.     

Identification of a SCAR marker linked to a recessive male sterile gene (Tems) and its application in breeding of marigold (Tagetes erecta)

In marigold, an F₂ segregation population of 167 plants was constructed from a cross of a line (M525A) carrying the male sterility trait × an inbred line (f53f). In line M525A, the male sterility trait was controlled by the recessive gene, Tems . The intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) techniques combined with bulked segregant analysis were used to develop markers linked to the trait. From a survey of the 38 ISSR primers and 170 SRAP primer combinations, only one SRAP marker that was closely linked to the target trait was identified and successfully converted into sequence characterized amplified region (SCAR) marker that was located within 2.4 cM from Tems locus. The marker was validated with five other two-type lines and in each case the male fertile plants were reliably identified. This SCAR marker therefore permits the efficient marker-assisted selection of male sterile individuals in breeding programmes of marigold and will greatly facilitate the breeding of F₁ cultivars.     

鲜食番茄茄红素基因的AFLP-SCAR分子标记

番茄红素(Lycopene)是功能性天然色素,因其高抗氧化功能而备受关注.为筛选高番茄红素品种,加速鲜食番茄分子标记辅助育种进程,本试验以高茄红素番茄骨干系F-516F8和低茄红素骨干系Nor作为试验材料,研究了鲜食番茄高茄红素基因的AFLP-SCAR分子标记.利用杂交F1代、自交F2代遗传群体,通过64对E/M引物组...     

Identification and characterization of RAPD and SCAR markers linked to anthracnose resistance gene in sorghum [Sorghum bicolor (L.) Moench]

Anthracnose, one of the destructive foliar diseases of sorghum growing in warm humid regions, is incited by the fungus Colletotrichum graminicola.The inheritance of anthracnose resistance was studied using the parental cultivars of Sorghum bicolor (L.) Moench, HC 136 (susceptible to anthracnose) and G 73 (anthracnose resistant). The F₁ and F₂ plants were inoculated with the local isolates of C. graminicola cultures. The F₂ plants showed a segregation ratio of 3 (susceptible): 1(resistant) indicating that the locus for resistance to anthracnose in sorghum accession G 73 segregates as a recessive trait in a cross to susceptible cultivar HC 136. RAPD (random amplified polymorphic DNA) marker OPJ 01₁₄₃₇ was identified as marker closely linked to anthracnose resistance gene in sorghum by bulked segregant analysis of HC 136 × G73 derived recombinant inbred lines (RILs) of sorghum. A total of 84 random decamer primers were used to screen polymorphism among the parental genotypes. Among these, only 24 primers were polymorphic. On bulked segregant analysis, primer OPJ 01 amplified a 1437 bp fragment only in resistant parent G 73 and resistant bulk. The marker OPJ 01₁₄₃₇ was cloned and sequenced. The sequence of RAPD marker OPJ 01₁₄₃₇ was used to generate specific markers called sequence characterized amplified regions (SCARs). A pair of SCAR markers SCJ 01-1 and SCJ 01-2 was developed using Mac Vector program. SCAR amplification of resistant and susceptible parents along with their respective bulks and RILs confirmed that SCAR marker SCJ 01 is at the same loci as that of RAPD marker OPJ 01₁₄₃₇ and hence, is linked to anthracnose resistance gene. Resistant parent G 73 and resistant bulk amplified single specific band on PCR amplification using SCAR primer pairs. The RAPD marker OPJ 01₁₄₃₇ was mapped at a distance of 3.26 cM apart from the locus governing anthracnose resistance on the sorghum genetic map by the segregation analysis of the RILs. Using BLAST program, it was found that the marker showed 100 per cent alignment with the contig{_}3966 located on the longer arm of chromosome 8 of sorghum genome. Therefore, these identified RAPD and SCAR markers can be used in the resistance-breeding program of sorghum anthracnose by marker-assisted selection.An erratum to this article can be found at     

利用多元回归分析鉴定与白桦纤维长度性状相关的分子标记

通过对天然白桦群体583株个体纤维长度测定，选取其中有代表性的100株个体，利用随机扩增多态性DNA标记（random amplification polymorphism DNA，RAPD）技术对其基因组差异分析，通过扩增条带与性状表现间的多元回归分析，筛选出与白桦纤维长度显著相关的分子标记。经过20个RAPD引物筛选，有6个引物的7个片段与纤维长度显著相关，其中片段“BFL”与纤维长度的相关系数为0．401，相关性达到5％的显著水平。对此片段进行克隆、测序后，成功转化成与长纤维性状相关的序列特征化扩增区域（sequence characterized amplified region,SCAR）标记，此标记对长纤维白桦的鉴定效率达80％。