甘蓝型油菜开花期全基因组关联分析及开花基因标记开发

随着南方稻-稻-油三熟制种植模式的推进和油菜机械化生产的要求,选育油菜早熟品种是现阶段中国南方油菜育种的重要研究目标。本研究对100份油菜品种进行简化基因组测序并调查开花时间,利用混合线性模型进行全基因组关联分析,进行SNP等位基因间的表型差异分析。结果表明,检测到69个SNP与开花时间显著关联,可以解释13.42%～23.28%的表型变异;检测到674个SNP等位基因间表型均值差异显著,其中有13个SNP位于与开花期相关的基因内;确认有5个SNP与开花时间性状显著有关,且这5个SNP等位基因间的表型差异达到了6.02～19.19 d;在显著关联位点上下游300 kb范围内筛选到的候选基因CLPS3和EBS可能对影响油菜开花时间起着重要作用,CLPS3和EBS特异性标记可以作为开花期的功能标记用于分子标记辅助育种。     

甘蓝型油菜角果长度性状的全基因组关联分析

角果长度是油菜重要的农艺性状,适度增加角果长度有利于扩大角果库容量,增加光合面积,提高油菜的籽粒产量。本研究利用Illumina60KSNP芯片对496份具有代表性的油菜资源进行基因型分析,考察群体在4个环境中的角果长度表型,利用MLM和GLM模型进行全基因组关联分析。结果表明, MLM模型检测到7个位点,联合解释25.01%的表型变异; GLM模型检测到25个位点,联合解释41.77%的表型变异。合并共同位点后得到27个位点,其中7个与前人报道的QTL重叠,其余20个是新鉴定的位点。效应最大的位点Bn-A09-p29991443位于A09染色体,在MLM和GLM模型中分别解释13.89%和12.86%的表型变异,携带其优异等位基因的材料平均角果长度增加0.89cm。同时,在该位点附近找到已克隆的油菜角果长度基因ARF18和BnaA9.CYP78A9。此外,在5个位点附近发现拟南芥已知角果长度基因GID1b、FUL、EOD3、DOF4.4和GA20ox1的同源拷贝。本研究结果有助于解析角果长度的遗传基础,为研究角果长度的调控机理,指导角果长度的遗传改良打下基础。     

基于高密度SNP标记对玉米籽粒相关性状的QTL定位

高产是玉米育种的主要目标之一,而玉米籽粒是产量构成最重要的因子。本研究以‘T32’与‘齐319’为亲本构建118份F2:3家系的双亲分离群体为材料,通过对不同家系的3个籽粒性状(粒长,粒宽和粒重)进行评价,结合高密度SNP标记的基因型鉴定结果,利用IciMapping 4.2软件中的复合区间作图法对粒长、粒宽和粒重进行了QTL定位。共检测到11个QTL,其中控制粒重、粒长和粒宽的QTL分别为2、2和7个,单个QTL可解释4.03%～11.34%的表型变异。研究结果将为玉米籽粒性状的遗传改良提供更多的遗传位点,也为后续的精细定位以及候选基因功能研究提供科学支持。     

玉米胚大小及其相关性状的QTL分析

胚大小在调控玉米籽粒营养组成及籽粒大小方面扮演着重要角色,解析玉米胚大小自然变异的遗传基础对玉米籽粒品质和产量的协同改良具有重要意义。本研究利用由普通玉米自交系B73和高油玉米自交系By804组配的RIL群体,结合高密度的SNP遗传图谱同时对玉米胚大小、籽粒大小和油分等15个性状进行QTL定位分析,共鉴定到82个QTL,包括38个胚大小性状QTL、23个籽粒大小性状QTL和21个油分性状QTL。每个QTL解释的表型变异范围为2.50%～26.32%,平均为7.94%。QTL的置信区间变幅为0.3～52.8 Mb,平均长度为11.6 Mb;置信区间小于5、10和20 Mb的QTL占所有QTL的比例分别为32.9%、58.5%和79.3%。对检测到的QTL位点进行共定位分析鉴定到15个QTL热点区域,其中10个热点区域至少调控胚大小、籽粒大小、油分等三类性状中的两类,5个QTL热点区域只控制一类性状。在上述热点区域内,鉴定到已克隆的控制玉米籽粒油分的主效基因DGAT1-2和影响籽粒发育的基因Dek15。这些结果为玉米籽粒品质改良提供了重要信息。     

双低杂交油菜‘丰油10号’干物质积累及养分吸收规律

为了解双低杂交油菜新品种‘丰油10号’养分吸收利用规律,以‘丰油10号’为材料,在河南油菜主产区进行田间试验,分析了油菜各器官干物质积累量以及成熟期氮、磷、钾、硫、硼养分的累积量。结果表明:油菜全株干物质积累量随生育进程的推进呈"S"形变化曲线,蕾薹期至花期呈快速增长趋势;油菜籽粒产量与苗期干物质积累量呈抛物线关系(R2=0.9798**),与开花期-成熟期干物质净增量呈显著线性关系(R=0.9879**)。籽粒是氮、磷的分配中心,分别占总吸收量的73.32%和75.16%;钾主要分布在茎枝和角果壳中,籽粒中钾素仅占总吸收量的21.31%;硫主要分布在角果壳和籽粒中;硼元素主要分布在茎枝和籽粒中,占总吸收量的69.89%;根中各养分分布均最少,分配比例也最低。新品种‘丰油10号’地上部分与全株干物质积累量均随生育时期的推进呈现"慢—快—慢"的变化曲线,根、茎、叶中的养分N和P在花期后会转移到籽粒中,因此,油菜花期前保证充足的养分供给是高产的前提。     

地下水位对油菜生长及产量的影响

长江流域油菜生产主要采用水稻油菜轮作种植模式,地下水位高,易产生湿害。本试验采用PVC筒,在油菜4个生育期分别进行0、30、60和90 cm的地下水位处理,然后恢复至60 cm水位,比较2个耐渍性不同的油菜品系在不同生育期、不同水位条件下的生长状况及产量,从而确定油菜各生育阶段的适宜地下水位,为南方稻茬油菜排水降湿管理提供依据。试验结果表明: (1)地下水位高低影响了油菜的根系发育、地上部生长及产量形成;(2)就产量而言,各生育期对水分的敏感性依次为蕾薹期、花期﹥苗期、角果发育成熟期;(3)油菜苗期适宜地下水位为30~90 cm,蕾薹期、花期为30~60 cm,角果发育成熟期为30~90 cm;(4)就全生育期而言,即使选用耐渍性较弱的品种,地下水位控制在30~60 cm时能满足油菜生长发育及产量形成需求。     

玉米开花期性状的QTL及杂种优势位点定位

开花期是玉米进化和适应过程中的重要性状,明确开花期杂种优势的遗传机制对培育适应不同生态区的优良玉米品种具有重要的意义。本研究利用以许178为受体,综3为供体构建的包含203个SSSL的单片段代换系群体及其与许178的测交群体,通过2年3个试点玉米开花期性状(散粉期、吐丝期和散粉至吐丝间隔)QTL和杂种优势位点(HL)分析,分别鉴定出40个开花期相关性状的QTL和37个开花期相关性状的HL。其中6个QTL和4个HL在3个地点被同时检测到。在所检测到的染色体区段中,11个区段同时包含调控开花期的QTL和HL。该研究为进一步解析玉米开花期遗传机制和开花期杂种优势的遗传机制提供了基础。     

陆地棉现蕾期和开花期性状的遗传分析

早熟性是作物育种的重要目标性状之一。以‘鲁棉研28号’和‘X1570’为亲本配制F1组合及其F2:3世代分离群体。利用‘鲁棉研28号’、‘X1570’、‘鲁棉研28号’בX1570’F1及F2:3群体进行多世代联合分析,以探究陆地棉现蕾期和开花期两个生育期相关性状的遗传规律。结果表明,现蕾期符合加性-显性-上位性多基因遗传模型,受多基因控制,且遗传率达85.00%;开花期符合1对显性主基因+加性-显性多基因的混合遗传模型,主基因和多基因的遗传率分别为87.69%和9.37%。主基因和多基因的加性效应值分别为2.543 1和3.419 6。相关系数分析表明,现蕾期、开花期都与棉花的果枝类型呈极显著相关。本研究结果可为现蕾期和开花期的QTL作图、长江流域棉花早熟育种工作提供一定的理论依据。     

甘蓝型油菜产量及其构成因素的QTL定位与分析

产量性状是复杂的数量性状, 对种子的单株产量及其构成因素(全株总有效角果数、每角粒数、千粒重)进行QTL定位和上位性分析,确定其在染色体上的位置及其遗传效应,可以探讨油菜杂种优势产生原因,提高育种中对产量性状优良基因型选择的效率,达到提高油菜产量的目的。在双低油菜细胞质雄性不育保持系1141B和双高恢复系垦C₁构建的F₂作图群体中,运用SRAP、AFLP和SSR三种标记技术构建了一个甘蓝型油菜(Brassica napus L.)的分子标记遗传连锁图谱。共包含244个标记,分布于20个主要连锁群、1个三联体上,图谱总长度为2 769.5 cM。采用Windows QTL Cartographer Version 2.0统计软件及复合区间作图法,对油菜单株产量及其3大构成因素进行QTL定位,共检测到QTLs 16个分布在9个连锁群上,其中第6和13连锁群最多,均有3个。单个QTL解释性状表型变异的0.38%～73.34%。对于同一性状,等位基因的增效作用既来自母本,亦源自父本;采用双向方差分析法对位点间互作及其上位性进行分析,检测到26对影响产量构成性状的上位性互作效应QTL,说明油菜基因组中存在大量控制产量的互作位点,油菜产量性状的上位性存在着多效性,上位性互作包括QTL与非QTL位点,其中以非QTL位点较多。一般互作位点的独立效应值较小,而互作的效应值显著增大,且一般超过两位点独立效应值之和。反映了控制产量性状基因的复杂性。上位性是甘蓝型油菜产量性状杂种优势的重要遗传基础。     

甘蓝型油菜株高、第一分枝高和分枝数的QTL检测及候选基因筛选

株高、分枝数及第1分枝高是油菜重要的农艺性状。本研究利用甘蓝型油菜GH06和P174杂交,F2通过单粒法连续自交至F11构建重组自交系群体,利用油菜60K芯片对该群体进行基因分型,构建高密度遗传连锁图谱。结果表明,该图谱包含2795个SNP多态性标记位点,总长1832.9 c M,相邻标记间平均距离为0.66 c M。在此图谱基础上采用复合区间作图法(CIM),检测到3个农艺性状的24个QTL。其中11个株高QTL分别位于A01、A06、A07、A08、A10和C06染色体,单个QTL解释5.00%~15.26%的表型变异;7个第1分枝高QTL分别位于A06、C05和C06染色体,单个QTL解释5.04%~12.99%的表型变异;6个分枝数QTL分别位于A03、A07、C01、C04和C06染色体,单个QTL解释5.95%~8.14%的表型变异。将156个拟南芥株高相关基因、10个拟南芥第1分枝高相关基因和148个拟南芥分枝数相关基因与QTL对应置信区间序列进行同源比较分析(E1E–20),分别找出了20个株高候选基因、3个第1分枝高候选基因以及12个分枝数候选基因。2个环境中在A07染色体上重复检测到的QTL置信区间检测到与株高相关的候选基因ATGID1B/GID1B和WRI1,A08染色体上重复检测到的QTL置信区间检测到SLR/IAA14和AXR2/IAA72个与株高相关的候选基因。在具有部分置信区间重叠的q2013FBH-C05-1和q2014FBH-C05-2区间均检测到第1分枝高候选基因PHT1;8,在A03和C06染色体上的QTL置信区间内,分别检测到4个分枝数候选基因,匹配E值介于0~3E–56之间。     

Quantitative trait loci (QTL) mapping of silique length and petal colour in Brassica rapa

Recombinant inbred lines (RILs) derived from a cross between Brassica rapa L. cv. ‘Sampad’, and an inbred line 3‐0026.027 was used to map the loci controlling silique length and petal colour. The RILs were evaluated under four environments. Variation for silique length in the RILs ranged from normal, such as ‘Sampad’, to short silique, such as 3‐0026.027. Three QTL, SLA3, SLA5 and SLA7, were detected on the linkage groups A3, A5 and A7, respectively. These QTL explained 36.0 to 42.3% total phenotypic variance in the individual environments and collectively 32.5% phenotypic variance. No additive × additive epistatic interaction was detected between the three QTL. Moreover, no QTL × environment interaction was detected in any of the four environments. The number of loci for silique length detected based on QTL mapping agrees well with the results from segregation analysis of the RILs. In case of petal colour, a single locus governing this trait was detected on the linkage group A2.     

Molecular and comparative mapping for heading date and plant height in oat

Selection of oat genotypes combining earliness and short plant height could stimulate oat cultivation worldwide. However, the mechanisms involved with the genetic control of heading date and plant height traits are not fully understood to date. This study aimed to identify genomic regions controlling heading date and plant height in two hulled by naked oat populations and to compare these genomic regions with that of other grass species. Recombinant inbred lines of each population and their parents were genotyped by a 6 K BeadChip Illumina Infinium array and assessed for heading date and plant height in two sowing dates. The quantitative trait loci (QTL) affecting these traits were detected by simple interval mapping. The two oat populations showed different genetic mechanisms controlling heading date. A major QTL was identified in one of the populations, mapped into the ‘Mrg33’ consensus linkage group from the current oat map. Two other QTL were detected into the ‘Mrg02’ and ‘Mrg24’ groups, in the second population. On the other hand, both populations presented the same genomic region controlling plant height. Six SNP markers, mapping on the same linkage group within each population, were associated with the trait, regardless the sowing date, explaining more than 20% of the phenotypic variation. Five of these six markers were mapped into three different linkage groups on the oat consensus map. Genomic regions associated with heading date and plant height in oat seem to be conserved in Oryza sativa L. and Brachypodium distachyon. Our results provide valuable information for marker-assisted selection in oats, allowing selection for earliness and plant height on early segregating generations.     

Identification of quantitative trait loci for flowering-related traits in the D genome of synthetic hexaploid wheat lines

The gene pool of Aegilops tauschii, the D-genome donor of common wheat (Triticum aestivum L.), can be easily accessed in wheat breeding, but remains largely unexplored. In our previous studies, many synthetic hexaploid wheat lines were produced through interspecific crosses between the tetraploid wheat cultivar Langdon and various A. tauschii accessions. The synthetic hexaploid wheat lines showed wide variation in many characteristics. To elucidate the genetic basis of variation in flowering-related traits, we analyzed quantitative trait loci (QTL) affecting time to heading, flowering and maturity, and the grain-filling period using four different F₂ populations of synthetic hexaploid wheat lines. In total, 10 QTLs located on six D-genome chromosomes (all except 4D) were detected for the analyzed traits. The QTL on 1DL controlling heading time appeared to correspond to a flowering time QTL, previously considered to be an ortholog of Eps-A ^m 1 which is related to the narrow-sense earliness in einkorn wheat. The 5D QTL for heading time might be a novel locus associated with wheat flowering, while the 2DS QTL appears to be an allelic variant of the photoperiod response locus Ppd-D1. Some of the identified QTLs seemed to be novel loci regulating wheat flowering and maturation, including a QTL controlling the grain filling period on chromosome 3D. The exercise demonstrates that synthetic wheat lines can be useful for the identification of new, agriculturally important loci that can be transferred to, and used for the modification of flowering and grain maturation in hexaploid wheat.     

Identification and mapping of a QTL for rachis internode length associated with cleistogamy in barley

General knowledge of the closed flowering trait, or cleistogamy, of barley is still limited. The relationship between cleistogamy and spike morphology characters was studied and linkage of cleistogamy genes with a highly significant quantitative trait locus (QTL) for rachis internode length on the long arm of chromosome 2H was detected. The mapping populations consisted of 129 doubled haploid lines of ‘Mikamo Golden’ × ‘Harrington’ and 150 F₂ plants of ‘Misato Golden’ × ‘Satsuki Nijo’. The phenotypic variance explained by this QTL accounted for 77.5% and 82.6% of the variance in rachis internode lengt, respectively, in these two populations. The peaks of the QTL coincided with the positions of the cleistogamy gene loci.     

Identification of coincident QTL for days to heading,spike length and spikelets per spike in <Emphasis Type="Italic">Lolium perenne</Emphasis> L.

Flowering time is a trait which has a major influence on the quality of forage. In addition, flowering and subsequent seed yields are important traits for seed production by grass breeders. In this study, we have identified quantitative trait loci (QTL) for flowering time and morphological traits of the flowering head in an F₁ mapping population in Lolium perenne L (perennial ryegrass), a number of which have not previously been identified in L. perenne mapping studies. QTL for days to heading (DTH) were mapped in both outdoor and glasshouse experiments, revealing three and five QTL for DTH which explained 53% and 42% of the total phenotypic variation observed, respectively. Two QTL for DTH were detected in both environments, although they had contrasting relative magnitudes in each environment. One QTL for spike length and three QTL for spikelets per spike were also identified explaining, a total of 32 and 33% of the phenotypic variance, respectively. Furthermore, the QTL for spike length and spikelets per spike generally coincided with QTL for days to heading, implying co-ordinate regulation by underlying genes. Of particular interest was a region harbouring overlapping QTL for days to heading, spike length and spikelets per spike on the top of linkage group 4, containing the major QTL for spike length identified in this population.     

Quantitative trait analysis of flowering time in spring rapeseed (B. napus L.)

The inheritance of flowering time trait in spring-type rapeseed (Brassica napus L.) is poorly understood, and the investigations on mapping of quantitative trait loci (QTL) for the trait are only few. We identified QTL underlying variation for flowering time in a doubled haploid (DH) mapping population of nonvernalization-responsive canola (B. napus L.) cultivar 465 and line 86 containing introgressions from Houyou11, a Chinese early-flowering cultivar in Brassica rapa L. Significant genetic variation in flowering time and response to photoperiod were observed among the DH lines from 465/86. A molecular linkage map was generated comprising three types of markers loci. QTL analysis indicated that flowering time is a complex trait and is controlled by at least 4 major loci, localized on four different linkage groups A6, A7, C8 and C9. These loci each accounted for between 9.2 and 12.56 % of the total genotypic variation for first flowering. The published high-density maps for flowering time mapping used different marker systems, and the parents of our crosses have different genetic origins, with either spring-type B. napus or B. rapa. So we cannot determine whether the QTL on the same linkage groups were in the same region or not. There was evidence of additive × additive epistatic effects for flowering time in the DH population. Epistasis existed not only between main-effect QTLs, but also between QTLs with minor effects. Four pair of epistasis effects between minor QTLs explained about 20 % of the genetic variance observed in the DH population. The results indicated that minor QTLs for flowering time should not be ignored. Significant genotypes × environment interactions were also found for the quantitative traits, and with significant change in the ranking of the DH lines in different environments. The results implied that FQ3 was a non-environment-specific QTL and may control flowering time by autonomous pathway. FQ4 were winter-environment-specific QTL and may control flowering time by photoperiod-pathway. Identification of the chromosomal location and effect of the genes influencing flowering time may hasten the development of canola varieties having an optimal time for flowering in target environments such as for high altitude areas, via marker-assisted selection.     

Mapping of QTL associated with waterlogging tolerance and drought resistance during the seedling stage in oilseed rape (Brassica napus)

Soil waterlogging and drought are major environmental stresses that suppress rapeseed (Brassica napus) growth and yield. To identify quantitative trait loci (QTL) associated with waterlogging tolerance and drought resistance at the rapeseed seedling stage, we generated a doubled haploid (DH) population consisting of 150 DH lines from a cross between two B. napus lines, namely, line No2127-17 × 275B F₄ (waterlogging-tolerant and drought-resistant) and line Huyou15 × 5900 F₄ (waterlogging-sensitive and drought-sensitive). A genetic linkage map was constructed using 183 simple sequence repeat and 157 amplified fragment length polymorphism markers for the DH population. Phenotypic data were collected under waterlogging, drought and control conditions, respectively, in two experiments. Five traits (plant height, root length, shoot dry weight, root dry weight and total dry weight) were investigated. QTL associated with the five traits, waterlogging tolerance coefficient (WTC) and drought resistance coefficient (DRC) of all the traits were identified via composite interval mapping, respectively. A total of 28 QTL were resolved for the five traits under control conditions, 26 QTL for the traits under waterlogging stresses and 31 QTL for the traits under drought conditions. Eleven QTL were detected by the WTC, and 19 QTL related to DRC were identified. The results suggest that the genetic bases of both waterlogging tolerance and drought resistance are complex. Some of the QTL for waterlogging tolerance-related traits overlapped with QTL for drought resistance-related traits, indicating that the genetic bases of waterlogging tolerance and drought resistance in the DH population were related in some degree.     

海拔、移栽期、打顶方式对翠碧一号烟叶产质量的影响

为研究福建省不同海拔区域不同移栽期和打顶方式对‘翠碧一号’烟叶品质的影响,采用田间随机区组试验对烟叶生长和品质进行研究。结果表明,在250 m海拔区域,随着移栽期和打顶期推迟,烟株生育期缩短、株高增加、发病率上升、产质量下降、化学成分协调性和感官质量下降;在650 m海拔区域,随着移栽期和打顶期推迟,烟株生育期缩短,农艺性状变化不显著,发病率变化不明显,产值量、化学成分协调性和感官质量呈先上升后下降的趋势。为了提高烟叶品质和产量,在福建省250 m海拔区域,‘翠碧一号’应提前移栽（1月10日）、初花打顶（10%中心花开发）;而在福建省650 m海拔区域,‘翠碧一号’应中期移栽（2月4日）、盛花打顶（40%中心花开发）。