两个玉米回交一代群体中opaque2基因单侧连锁累赘的SSR分析

为了在回交一代既能较好地消除靶基因的遗传连锁累赘，又能在进行背景选择的基础上选择到轮回亲本遗传背景回复率较高的单株，本研究构建了单株数分别为1 416和1 627的SCBC₁F₁和HCBC₁F₁两个不同遗传背景的回交群体。结合玉米叶片DNA大量、快速提取法，使用5个与opaque2（o2）基因连锁并已定位的SSR标记，在SCBC₁F₁与HCBC₁F₁群体中，对o2基因单侧的连锁累赘进行SSR分析，并与Ribaut（2002）的计算机模拟结果进行比较，结果表明，本研究获得的结果优于相应的计算机模拟结果。并对分子标记辅助选择体系在玉米回交育种中应用的若干理论问题进行了讨论，认为在实际应用MAS程序中，不仅要重点考虑最终决选的单株数（N_i），而且还应该考虑到实际有足够后代的中选单株数（N_j）。另外，结合预期要消除连锁累赘的图距，制备足够大的BC₁F₁群体是十分必要的。     

一个水稻落粒性基因SH1的SSR标记定位

以籼稻品种93-11为轮回亲本,与粳稻品种日本晴杂交并回交的高世代分离群体为研究材料,选用104个多态性的SSR标记对水稻的落粒性基因进行定位。结果表明,在BC₄F₂群体中,6个标记的基因型来自于日本晴;在BC₄F₃定位群体中,难落粒植株数与易落粒植株数的分离比例为3:1,落粒性受1对显性基因控制,命名为SH1;分子标记与落粒性共分离分析将SH1定位在SSR标记RM5389和RM1068、RM1387之间,与3个标记的遗传距离分别为0.7cM、5.5cM和13.1cM,此结果为该基因的分子标记辅助选择奠定了基础。     

玉米o16基因回交渗入o2系的分子标记辅助选择

在高赖氨酸玉米育种中,主要是以玉米opaque-2(o2)突变体作供体回交转育培育亲本材料,再行培育高赖氨酸杂交种。但是,迄今培育的o2玉米系及其杂交种的籽粒赖氨酸含量约为0.4%,不能满足食用和饲用的需求。为了提高o2玉米的赖氨酸含量,本研究利用一个新的高赖氨酸突变基因opaque-16(o16)的载体QCL3021作供体,o2玉米系太系19为受体,将o16基因回交渗入o2玉米系。在回交的每一世代及随后的自交世代,用o2基因内的SSR标记umc1066和o16基因的连锁SSR标记umc1141进行前景选择,再对中选单株进行全基因组SSR标记的背景选择,最后用染料结合赖氨酸法测定籽粒赖氨酸含量,以便保证筛选出遗传背景恢复率和赖氨酸含量均高的目标单株。在BC₂F₄代,获得携带o2和o16基因的家系17个,其遗传背景与o2玉米系相当(恢复率为92%~95%),赖氨酸含量为0.469%~0.599%。其赖氨酸含量比普通玉米平均提高约122.63%;比高值亲本太系19(o2o2)平均提高约22.33%,增幅为6.11%~35.52%;比低值亲本QCL3021(o16o16)平均提高约65.86%,增幅为43.87%~83.74%。表明采用标记辅助选择技术将o16基因回交导入o2玉米,能有效提高玉米籽粒的赖氨酸含量,对高赖氨酸玉米的遗传改良和育种具有重要意义。     

一个水稻抽穗期主基因Hd(t)的遗传分析及分子定位

从缙恢10/R21的杂种后代中发现了一个抽穗期稳定遗传的迟熟恢复系N91(110~114 d),以早熟不育系金23A(89~94 d)作为杂交和回交亲本,获得的F₂和BC₁F₁群体抽穗期均表现双峰分布,χ²检测表明其抽穗期受一对主基因控制,暂命名为Hd(t)。在400多对SSR引物中筛选出5对在早熟基因池和迟熟基因池中表现差异的引物,进行单株验证,用回交群体进行基因定位,发现位于第7染色体长臂末端的SSR标记RM1364和RM3555与Hd(t)连锁,遗传距离分别为32.7 cM和22.5 cM。在目标区域进一步合成8对SSR引物,将Hd(t)基因定位在RM22143与第７染色体末端之间,与RM22143相距12.9 cM。该结果为Hd(t)基因的精细定位、分子标记辅助育种和基因克隆奠定了基础。     

玉米抗南方型锈病基因共分离分子标记的研究

本文以玉米(Zea mays L.)抗南方型锈病自交系P₂₅和感病自交系F₃₄₉的F₁与F₃₄₉回交,并连续回交所得BC₅代的F₃₄₉抗病近等基因系（nearly isogenic lines,NILs）为材料,以RGA（resistance gene analogs）的方法,在抗病自交系中克隆出一条321 bp的特异性DNA条带。根据序列比对的结果,设计出新的引物作为分子标记,在96株BC₇、BC₄F₅NILs及其亲本和杂种F₁群体上进行鉴定,其中94株的田间抗感结果与分子检测结果一致,选择的有效率达97.9%。此标记扩增效果清晰,可重复性强,在抗玉米南方型锈病的分子标记辅助育种研究中有应用价值。     

大豆脂肪及脂肪酸组分含量的QTL定位

脂肪及脂肪酸组分的改良是大豆油脂品质育种的主要方面。本研究旨在构建遗传图谱,定位大豆脂肪及脂肪酸组分的QTL,为大豆油脂品质育种提供参考。以Essex×ZDD2315的114个BC₁F₁单株为作图群体,构建了250个SSR标记和1个形态标记,具有25个连锁群的遗传图谱,覆盖大豆基因组2 963.5 cM,平均每个连锁群上10.0个标记,标记平均间距11.8 cM。用BC₁F₃家系3个重复的表型平均值代表相对应的BC₁F₁单株表型值,采用Win QTL Cartographer 2.5复合区间作图法（CIM）检测到18个控制脂肪及脂肪酸组分含量的QTL,位于9个不同的连锁群上,表型贡献率为9.6%~34.5%;多区间作图法（MIM）检测到与CIM区间相同的7个QTL（fat-1, pal-1, st-1, ole-1, lin-1, lin-4和lio-2）,区间相近的2个QTL（ole-4和lin-5）,位于6个不同的连锁群上,表型贡献率为8.2%~39.3%。CIM法检测到的其他9个QTL有待进一步验证。大豆脂肪及脂肪酸组分含量的主效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,育种中既要注意主效QTL的利用,又要考虑微效多基因的积聚。     

一个新的抗玉米矮花叶病基因的发现及初步定位

由SCMV引起的矮花叶病是我国的主要玉米病害之一, 鉴定和发掘新的抗病基因对于玉米抗病遗传育种具有重要意义。以抗病自交系海9-21和感病自交系掖478杂交的一个BC₂F₃群体为试验材料, 通过人工接种矮花叶病毒进行抗病性鉴定, 发现该分离群体中抗病植株与感病植株数符合1∶3的分离比例, 推测其抗病基因是由1对隐性基因控制。抗感池和SSR标记连锁分析表明, 存在一个新的玉米矮花叶病隐性抗病基因(或等位基因), 将该基因命名为scm3。scm3基因来源于抗病玉米自交系海9-21, 位于第3染色体短臂3.04~3.05区域, 在SSR标记umc1965和bnlg420之间, 遗传距离分别为45.7 cM和6.5 cM。连锁的标记还有umc1307、umc2265、bnlg2241和umc2166, 它们与scm3之间的遗传距离分别是8.3、13.3、15.5和19.7 cM, 这些SSR标记与scm3基因在染色体上的排列顺序为umc1965—scm3—bnlg420—umc1307—umc2265—bnlg2241—umc2166。     

玉米种子休眠性的QTL定位

选用两个种子休眠性差异较大的普通玉米自交系R08与A318组配的F2群体共331个单株,构建了包含137个SSR标记的分子遗传连锁图谱,覆盖玉米基因组2 076.7 cM,平均图距15.2 cM。采用复合区间作图法对F_2:3家系种子休眠性数据进行分析,共检测到7个QTL,分别位于玉米第1、3、5和10染色体上。7个QTL的贡献率在2.45%~26.     

玉米抗纹枯病QTL定位

以玉米自交系R15（抗）×掖478（感）的229个F₂单株为作图群体,构建了包含146个SSR标记位点的遗传连锁图谱,全长1 666 cM,平均图距11.4 cM。通过麦粒嵌入法对F2:4群体进行人工接种纹枯病菌,并以相对病斑高为病级划分标准鉴定了玉米纹枯病的抗性。用复合区间作图法分析抗病QTL及遗传效应,共检测到9个抗性QTL,分布于第1、2、3、4、5、6和10条染色体上,单个QTL可解释表型方差的3.72%~7.19%,其中有2个QTL位于染色体6.01抗病基因簇附近。     

水稻抗白叶枯病基因Xa23的RFLP标记定位及其STS标记的转化

用水稻抗白叶枯病基因Xa23的近等基因系CBB23与其感病轮回亲本金刚30（JG30）杂交,构建了包含2562个单株的F₂作图群体。用水稻白叶枯病广致病菌系P6进行抗性鉴定表明,F₂植株抗感分离比严格符合3：1。根据日本水稻基因组计划RGP水稻高密度图谱上的RFLP探针对F₂群体中的145个感病单株进行RFLP检测和连锁分析,获得了6个与Xa23紧密连锁的RFLP分子标记。其中RFLP标记C1003A靠着丝粒一侧,与Xa23的遗传图距为0.4cM,为Xa23的图位克隆奠定了重要基础。并将标记C1003A成功地转化为STS标记,为分子标记辅助选择育种（MAS）提供了方便有效的分子标记。     

籼稻背景的单片段代换系群体的构建

以国内、外12个水稻品种（包括8个籼稻品种和4个粳稻品种）为供体亲本，利用回交和微卫星标记辅助选择相结合的方法构建了一个以籼稻品种“华粳籼74”为遗传背景的单片段代换系（SSSL）群体。该群体由260个不同的SSSL组成，其代换片段分布在水稻的12条染色体上，长度分布于0.2～109.9 cM，平均长度为19.9 cM；代换片段的总长度为5 166.0 cM，相当于水稻基因组总长度的3.4倍，代换片段在水稻基因组上的覆盖率为83.8%。这些SSSL的代换片段包含了水稻基因组丰富的等位基因变异，将在基因（QTL）的定位、克隆、功能分析及水稻分子育种中具有重要的利用价值。     

短季棉早熟性的分子标记及QTL定位

以两个陆地棉品种中棉所36×TM-1的207个F2单株为作图群体,筛选出73个多态性引物,25个SSR标记、35个RAPD标记和13个SRAP标记,构建了第一张以研究短季棉为主的包含43个标记,标记间的最小遗传距离为11.8 cM,最大遗传距离为48.9 cM,总长1174.0 cM的遗传连锁图谱,覆盖棉花基因组总长度的23.48%。检测到与短季棉早熟性状相关的12个QTLs,其中有8个QTLs呈簇分布在LG1连锁群上,找到对表型变异的贡献率在30%以上与全生育期、霜前花率和开花期有关的QTL各1个。     

玉米幼胚培养能力性状QTL分析

以18-599（红）和R15组配的F₂群体,构建了含88对SSR标记的玉米遗传连锁图谱,覆盖基因组1 636.6 cM,标记间平均距离为18.6 cM。结合幼胚组织培养试验将幼胚发生胚性愈伤组织诱导率和胚性愈伤组织绿苗再分化数作为反映玉米培养过程中胚性愈伤组织诱导力和胚性愈伤组织绿苗再分化力的性状指标,以此调查玉米幼胚培养能力。利用复合区间作图法进行QTL分析,共检测到5个与胚性愈伤组织诱导率有关的QTL,位于第1、3、7和8染色体上,可解释5.25%～23.4%的表型变异;检测到1个与胚性愈伤组织绿苗发生数有关的QTL,位于第3染色体上,可解释表型变异率为13.42%。     

An SSR-based molecular genetic map of cassava

Summary Microsatellites or simple sequence repeats (SSR) are the markers of choice for molecular genetic mapping and marker-assisted selection in many crop species. A microsatellite-based linkage map of cassava was drawn using SSR markers and a F₂ population consisting of 268 individuals. The F₂ population was derived from selfing the genotype K150, an early yielding genotype from an F₁ progeny from a cross between two non-inbred elite cassava varieties, TMS 30572 and CM 2177-2 from IITA and CIAT respectively. A set of 472 SSR markers, previously developed from cassava genomic and cDNA libraries, were screened for polymorphism in K150 and its parents TMS 30572 and CM 2177-2. One hundred and twenty two polymorphic SSR markers were identified and utilized for linkage analysis. The map has 100 markers spanning 1236.7 cM, distributed on 22 linkage groups with an average marker distance of 17.92 cM. Marker density across the genome was uniform. This is the first SSR based linkage map of cassava and represents an important step towards quantitative trait loci mapping and genetic analysis of complex traits in M. esculenta species in national research program and other institutes with minimal laboratory facilities. SSR markers reduce the time and cost of mapping quantitative trait loci (QTL) controlling traits of agronomic interest, and are of potential use for marker-assisted selection (MAS).     

大豆灰斑病1号生理小种抗性基因的SSR标记

针对中国大豆灰斑病1号生理小种,以抗所有生理小种的品系东农40566为母本,以感所有生理小种的品种东农410为父本配制杂交组合,杂交得到F2代后连续自交3代得到F5代群体。该群体经人工接种灰斑病1号生理小种后,利用BSA法对500个SSR标记进行筛选,其中3个标记Satt565、SOYGPATR和Satt396在抗、感池间表现出稳定的多态性,并且在F2代个体中表现出抗性与多态性协同分离的趋势。3个标记与抗性基因的连锁顺序为Satt565—SOYGPATR—Hrcs1—Satt396,它们与抗性基因的连锁距离分别为12.7cM、6.5cM、14.7cM。推测抗大豆灰斑病1号生理小种的基因可能位于C1连锁群上。     

Genetic linkage map of Chinese native variety faba bean (Vicia faba L.) based on simple sequence repeat markers

Simple sequence repeat (SSR) marker is a powerful tool for construction of genetic linkage map which can be applied for quantitative trait loci (QTL) and marker‐assisted selection (MAS). In this study, a genetic map of faba bean was constructed with SSR markers using a 129 F₂ individuals population derived from the cross of Chinese native variety 91825 (large seed) and K1563 (small seed). By screening 11 551 SSR primers between two parents, 149 primer pairs were detected polymorphic and used for F₂ population analysis. This SSR‐based genetic linkage map consisted of 15 linkage groups with 128 SSR. The map encompassed 1587 cM with an average genetic distance of 12.4 cM. The genetic map generated in this study will be beneficial for genetic studies of faba bean for identification of marker‐locus‐trait associations as well as comparative mapping among faba bean, pea and grasspea.     

Quantitative trait loci for quality and agronomic traits in two advanced backcross populations in oat (Avena sativa L.)

Using the advanced backcross quantitative trait loci (AB‐QTL) strategy, we successfully transferred and mapped valuable allelic variants from the high β‐glucan (BG) accession IAH611 (PI 502955), into the genome of cultivar ‘Iltis’. By backcrossing one BC₁F₁ plant to ‘Iltis’, we developed two BC₂F_2‐6 populations A and B, comprising 98 and 72 F₂‐individuals, respectively. Genotyping of BC₂F₂ individuals with predominantly AFLP markers resulted in 12 linkage groups with a map size of 455.4 cM for Population A and 11 linkage groups with a map size of 313.5 cM for Population B. Both populations were grown at three sites in Germany over a three‐year period. Individuals were then phenotyped for 13 traits including grain yield (YD) and β‐glucan content (BG). QTL analysis via stepwise regression detected a total of 33 QTLs, most of which were clustered in three linkage groups. Two dense linkage groups A1 and B13 were found to be putatively homologous to groups KO_6 and KO_11 of the ‘Kanota’/‘Ogle’ map, respectively.     

Rapid and high-precision marker assisted backcrossing to introgress the <Emphasis Type="Italic">SUB1</Emphasis> QTL into BR11, the rainfed lowland rice mega variety of Bangladesh

Flooding is one of the major hazards of rice production for the rainfed lowland rice ecosystem, and tolerant cultivars are urgently needed to help protect farmers from submergence damage. A quick and efficient strategy was implemented to introgress SUB1, a major QTL for submergence tolerance, into a rainfed lowland mega variety BR11 of Bangladesh by only two backcrosses and one selfing generation. In marker-assisted backcrossing (MABC), one tightly-linked simple sequence repeat (SSR) and two gene-based markers, four flanking SSR and 116 background SSR markers were used for foreground, recombinant and background selection, respectively, in backcrosses between a SUB1 donor IR40931-33-1-3-2 and BR11. BR11-Sub1, identified in a BC₂F₂ plant, possessed BR11 type SSR alleles on all fragments analyzed except the SUB1 QTL. The introgression size in BR11-Sub1 was 800 Kb indicating approximately 99.8% identity to BR11. BR11-Sub1 along with other introgression lines showed submergence tolerance similar to the tolerant parent. Yield, yield-component parameters and grain physico-chemical properties showed successful recovery of the BR11 traits in BR11-Sub1, with yield potential ranging from 5.2 to 5.6 t/ha, not significantly different from the recurrent parent mega variety BR11. Producing a large number (~1000) of backcross F₁ plants was considered essential to achieve recombination on both sides of the gene, limiting linkage drag with only two backcrosses. A large number of background markers ensured proper recovery of the recurrent parent genome in the BC₂F₂ generation. The study demonstrates a rapid and highly precise strategy to introgress a major QTL by BC₂F₂ generation into a modern rice variety using an unadapted donor. The variety can replace BR11 on more than 2 million of ha in Bangladesh and provide major increases in rice production.     

利用3个F_2群体整合高密度栽培种花生遗传连锁图

遗传图谱的构建及整合是开展花生分子育种研究的基础,利用多个作图群体整合遗传图谱是解决图谱标记密度低的有效途径。本研究采用基于锚定SSR标记的作图策略,构建3个F_2群体3张遗传连锁图,利用Join Map 3.0软件整合图谱,获得一张包含20个连锁群、792个位点、总遗传距离为2079.50 c M,标记间平均距离为2.63 c M的整合图谱,各连锁群标记数在20~66个之间,遗传距离在59.10~175.80 c M之间。将3个分离群体中检测到的与荚果及种子大小相关的QTL区段与整合连锁图的标记比较发现,各群体中检测到的位于各染色体上的QTL在整合图谱中都能出现,有些QTL标记区间在整合图谱中存在更多的标记,为今后利用这些标记进行精细定位奠定了基础。     

The first genetic maps of cashew (Anacardium occidentale L.)

Cashew (Anacardium occidentale) is a widespread tropical tree crop that is grown primarily for its nuts and has a global production of over 2 million Mt. In spite of its economic importance to many countries, however, no linkage map containing STS anchor sites has yet been produced for this species. This is largely attributable to a prolonged juvenile phase of the tree (limiting mapping to F₁ progenies) and difficulty in effecting sufficient hand-pollinations to create mapping populations of effective size. Here, we produce an F₁ mapping population of 85 individuals from a cross between CP 1001 (dwarf commercial clone) and CP 96 (giant genotype), and use it to generate two linkage genetic maps comprising of 205 genetic markers (194 AFLP and 11 SSR markers). The female map (CP 1001) contains 122 markers over 19 linkage groups and the male map (CP 96) comprises 120 markers assembled over 23 linkage groups. The total map distance of the female map is 1050.7 cM representing around 68% genome coverage, whereas the male map spans 944.7 cM (64% coverage). The average map distance between markers is 8.6 cM in the female map and 7.9 cM in the male map. Homology between the two maps was established between 13 linkage groups of the female map and 14 of the male map using 46 bridging markers that include 11 SSR markers. These maps represent a platform from which to identify loci controlling economically important traits in this crop.