与SP1互作的水稻穗顶部退化基因qPAA3的精细定位

【目的】水稻顶部小穗退化减少了单穗的总枝梗数和总粒数,严重影响单株产量,是水稻生产上的一个不利性状。因其遗传基础复杂,受环境影响较大,控制顶部小穗退化的相关基因克隆研究报道极少,该不利性状发生的分子机制及其遗传网络还不得而知。对顶部小穗退化基因进行精细定位,可为穗顶部基因的克隆奠定基础;开发的紧密连锁分子标记,也可以运用于分子育种实践,对这一不利性状进行早期识别和淘汰。【方法】首先对小穗突变体sp进行精细定位。用sp分别与粳稻品种ITA182和籼稻品种J160杂交构建2个遗传定位群体。为了研究不同穗退化突变体之间的关系,再以小穗突变体sp和穗顶部退化材料05261杂交,获得了农艺性状稳定的拟双突变体（表型与sp相似）。通过连续自交,纯合拟双突变体的遗传背景。再以高代的拟双突变体为非轮回亲本,穗顶部正常品种IRAT129为轮回亲本,构建含有双突变体的BC₁F₂亚群体。其中一个亚群体14C2017既表现单基因的穗退化性状分离,又出现小穗和穗顶部退化的双突变体表型,被用作顶部小穗退化基因的精细定位材料。【结果】水稻小穗性状是由1对隐性基因（sp）控制的。利用混池方法将SP(t)初步定位于第11染色体分子标记RM26281与RM7391之间;利用新开发的60对SSR分子标记,将其定位在标记sc50和sc66之间。在此区间内设计引物,最终将SP(t)定位在标记sc24和sc66之间,物理距离为54.3 kb的范围内。测序结果表明,突变体在该区间内有15.03 kb的大片段缺失,导致基因SP1的编码序列缺失。对拟双突变体的表型分析表明,sp与一个穗顶部退化基因存在互作。利用亚群体14C2017作为克隆与SP1互作基因的遗传分离群体,利用分布于全基因组的239对引物,筛选出在拟双突变体和IRAT129之间有多态的引物114对,将目标基因精细定位于第3染色体SSR标记RM6929和RM1319之间,物理距离为97.3 kb范围内,该候选基因属于早先报道的QTL--qPAA3。【结论】水稻sp的小穗性状是由基因SP1引起的缺失突变。与SP1互作的qPAA3定位于第3染色体SSR标记RM6929和RM1319之间,物理距离为97.3 kb的范围内。     

利用BSA法发掘野生大豆种子硬实性相关QTL

【目的】野生大豆的硬实性是大豆遗传改良利用中的重要限制因素。利用BSA法发掘与大豆种子硬实性相关的QTL,为野生大豆在大豆遗传改良中的合理利用奠定基础。【方法】利用栽培大豆中黄39与野生大豆NY27-38杂交构建F₂和F₇分离群体,从每个单株选取整齐一致的种子,取30粒种子置于铺有一层滤纸的培养皿中,加入30 mL蒸馏水,25℃培养箱中暗处理4 h,设3次重复,分别统计每个培养皿中正常吸胀和硬实种子数。在F₂群体中,选取22个正常吸胀单株（吸胀率＞90%）和16个硬实单株（吸胀率＜10%）;在F₇群体中,选取20个完全吸胀单株（吸胀率=100%）和20个完全硬实单株（吸胀率=0%）,单株DNA等量混合,分别构建2个吸胀和2个硬实DNA池。利用259对在亲本间有多态性的SSR标记对吸胀和硬实DNA池进行检测,筛选在吸胀和硬实DNA池间表现多态性的SSR标记;用192个SSR标记检测F₇分离群体,构建遗传图谱,利用复合区间作图法定位大豆硬实相关QTL。【结果】利用F₂个体构建的吸胀和硬实DNA池,在第2染色体16.3 Mb区间和第6染色体23.4 Mb区间分别检测到10个和8个在两池间有差异的SSR标记。利用这些标记检测F₂群体,将第2染色体的QTL定位于Satt274与Sat_198间的276.0 kb区间,该区间包括已克隆的大豆硬实基因GmHs1-1,解释17.2%的表型变异。第6染色体的QTL位于标记BARCSOYSSR_06_0993与BARCSOYSSR_06_1068间,可解释17.8%的表型变异。利用F₇株系构建的吸胀和硬实DNA池,在第2（27.4 Mb区间）、6（27.8 Mb 区间）和3染色体（18.2 Mb区间）分别检测到11个、9个和4个在两池间有多态性的SSR标记。利用F₇群体构建包括192个SSR标记、覆盖2 390.2 cM的遗传图谱,共检测到3个硬实相关QTL,其中第2染色体定位到的QTL位于标记Satt274与Sat_198间,可解释23.3%的遗传变异。第6染色体定位到的QTL位于标记Sat_402与Satt557之间,可解释20.4%的表型变异。在第3染色体标记Sat_266与Sat_236间发现一个可以解释4.9%表型变异的QTL,与BSA法检测的结果相符。【结论】利用BSA法可以检测到传统遗传作图定位的所有与硬实性相关的QTL,证明BSA法发掘大豆种子硬实性主要QTL的高效性。     

棉花早熟性QTL定位研究进展

运用各种分子标记技术和数量性状基因的定位方法,将控制棉花早熟性状的基因定位到相应的遗传连锁图谱或染色体上,为目前及今后短季棉育种提供了理论依据。综述了近年来棉花早熟性QTLs定位研究的最新进展,并对研究中存在的主要问题进行了分析和展望。     

罗非鱼耐盐数量性状位点(QTL)定位研究

采用190个微卫星分子标记对罗非鱼282个全同胞个体进行数量性状位点(QTL)定位,获得15个罗非鱼耐盐QTL,其中st-c、mst-a可分别解释19.2%的两性耐盐差异和19.1%的雄性耐盐差异,XY性别决定位点、钠钾氯协同转运蛋白2(NKCC2)基因分别定位于st-c、mst-a的峰值处。研究结果表明,罗非鱼海水养殖中采用全雄群体可能具有一定的耐盐优势,NKCC2基因可作为重要的候选基因用于耐盐罗非鱼选育。     

Mapping QTLs for Yield Stability in Durum Wheat Grown under Different Water Regimes

Among the most important Mediterranean annual crops, durum wheat is widely grown in drought-prone areas. Therefore, improving water-use efficiency （WUE） of durum wheat represents a major breeding goal. IDu-WUE （Improving Durum wheat for Water Use Efficiency and yield stability through physiological and molecular approaches） is a collaborative project among public and private research centres in Italy, Spain and WANA （West Asia and North Africa） countries （Morocco, Tunisia, Syria and Lebanon） funded by the European Union aimed at investigating the genetic variation for WUE and yield stability in durum wheat grown in Mediterranean droughtprone areas. During the first year of the project, a number of morpho-physiological traits （e.g. early vigour, flowering time, leaf rolling, number of fertile tillers, etc.）, WUE, WUE-related traits （e.g. carbon isotope discrimination, canopy temperature, chlorophyll fluorescence, etc.）, yield and its components have been investigated in a RIL population （249 lines） and a collection of ca. 190 durum wheat accessions characterized by a high level of linkage disequilibrium （Maccaferri et al., 2005）,     

Improving Drought Tolerance of Rice by Designed QTL Pyramiding

Drought is the most important factor limiting rice yields in the rainfed areas of Asia. To overcome the problem, we developed a new strategy ＇designed QTL pyramiding＇ to more efficiently develop drought tolerant （DT） rice cultivars. In the first phase of this strategy, we developed large numbers of introgression lines （ILs） in elite backgrounds using backcross （BC） breeding, each of which carries multiple genomic segments for improved DT from a known donor. Then, we genotyped all ILs with SSR markers to track the genomewide pattern of introgression in the DT ILs of 16 BC populations derived from crosses between 2 recipients （IR64 and Teqing） and four different donors. X^2 tests and linkage disequilibrium analyses of introgression in the DT ILs revealed significant frequency shifts at many loci across the genome and non-random associations at the multilocus level as a result of selection for DT in the ILs, which led us to the discovery of putative genetic networks underlying DT in the ILs. The networks each containing all DT loci detected in ILs from a BC population showed some interesting features.     

Mining Applicable Elite Alleles of Growth Duration,Plant Height and Panicle Number per Plant by Conditional QTL Mapping in Japonica Rice

Unconditional and conditional QTL mapping were conducted for growth duration (GD), plant height (PH) and effective panicle number per plant (PN) using a recombinant inbred line (RIL) population derived from a cross between two japonica rice varieties Xiushui 79 and C Bao. The RIL population consisted of 254 lines was planted in two environments, Nanjing and Sihong, Jiangsu Province, China. Results showed that additive effects were major in all of QTLs for GD, PH and PN detected by the two methods, and the e...     

大豆脂肪酸含量的QTL分析

利用Charleston(♀)×东农594(♂)的F14和F15代永久自交系群体154个单株后代,在2年3点条件下用气相色谱法测得其籽粒5种脂肪酸的含量,利用Win QTL Cartographer2.5复合区间作图法(CIM)进行QTL分析。结果共检测到47个相关的QTL,分布在13个连锁群上。多年多点同时检测到的QTL共有15个,其中控制软脂酸性状的2个,包括qPal-C2-2和qPal-A1-1;控制硬脂酸性状的4个,包括qSt-B1-1、qSt-B1-2、qSt-D1a-1和qSt-C2-1;控制油酸性状的3个,包括qOle-B2-1、qOle-G-1和qOle-H-1;控制亚油酸性状的有2个,包括qLin-C2-1和qLin-H-1;控制亚麻酸性状的4个,包括qLino-B1-1、qLino-C2-1、qLino-D1b-1和qLino-J-1。这些QTL的一致性较高,为特异脂肪酸含量标记辅助育种奠定了基础。大豆脂肪酸含量的主效QTL数量不多,效应大的不多,可能还受许多未能检测出来的微效基因控制。     

Precision of estimated QTL positions in granddaughter designs using combined haplotype sharing TDT and linkage analysis

The aim of this study was to develop the linear haplotype sharing transmission disequilibrium test (LHS-TDT) method and combine this method with the simple regression method to estimate the precision of QTL positions in granddaughter designs. This precision was determined by Monte Carlo simulation in granddaughter designs. A single bi-allelic QTL at the midpoint of a linkage group and 26 markers with 1 cM intervals and with two alleles each were simulated. Three linear models, (i.e. the simple regression model, the linear haplotype sharing TDT method and the combination of these two models) were compared. The mean of absolute differences (A) between the estimated and true QTL position of each method was considered for six different scenarios consisting of combinations of a number of markers and the most frequent haplotypes. The mean of A, using the simple regression method, was 4.38 centimorgan (cM). The means of A using the LHS-TDT method were less than the simple regression method in all scenarios and ranged from 1.86 to 3.82 cM depending on the scenario. The mean of A using the combined method was more than the LHS-TDT method and less than the simple regression method. The means of A using the combined method ranged from 2.32 to 4.36 cM. Therefore, for populations similar to those population simulated in this study, the LHS-TDT was better than the simple regression method and the combined method for precision of estimated QTL position in granddaughter designs.