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。     

玉米抗灰斑病基因的分子标记

玉米灰斑病是我国玉米生产中的重要病害,培育和种植抗病品种是防治此病的有效途径.1年内在3个灰斑病重发区选取64份我国常用玉米自交系进行了抗灰斑病鉴定,分别筛选出高抗、抗病和中抗自交系2份、15份和12份.采用集团混合分组分析法,分别以10个抗病和10个感病玉米自交系基因组DNA构建抗病基因池和感病基因池.从100对AFLP引物组合中筛选出54对在抗感基因池间呈多态性的引物,进一步在组建抗池和感池的20份自交系之间检测到8个多态性片段.通过片段的回收、克隆和测序,将其中的P51M38-100扩增片段转化为SCAR标记(SCAR-100).利用SCAR-100标记分析64份玉米自交系的基因型,结合田间抗病和感病表型进行相关分析,卡方测验表明SCAR-100标记与抗灰斑病显著相关.采用Mo17×黄早四群体(190个F2单株)和X178×B73群体(181个F8家系),结合已构建的SSR和AFLP标记连锁图谱,均将SCAR-100标记定位于第3染色体上,分别位于SSR标记umc1399-bnlg1754和umc1320-bnlg1754之间.开发抗病基因的分子标记可为分子标记辅助育种提供基础.     

玉米第10染色体大刍草单片段代换系的构建及穗长QTL分析

为了构建玉米第10染色体大刍草单片段代换系群体并进行穗长QTL的筛选。以玉米野生近缘种墨西哥类玉米为供体亲本,以玉米自交系郑58为受体亲本和轮回亲本连续进行多代回交。利用BC_7F_1至BC_9F_1连续回交群体,选用45对在供体和受体亲本间有明显多态性差异的引物进行SSR分子标记辅助选择,构建以墨西哥类玉米为供体亲本的玉米第10染色体单片段导入系群体。对穗长性状QTL在第10染色体上位置进行初步定位。经过连续多代回交和SSR标记跟踪检测,获得了以郑58为遗传背景的墨西哥类玉米导入系群体,共检测到单片段代换系材料107份,代换片段平均长度为25.52~45.97 c M,总长度为944.15~1 884.74 c M,对第10染色体覆盖率为70.38%~89.65%。在BC_9F_1回交群体中鉴定了4个来自墨西哥类玉米并位于第10染色体的穗长QTL,初步定位于umc2528、phi054、bnlg1360和umc1877标记附近。结果表明,利用大刍草作为供体亲本可以获得大量玉米第10染色体单片段代换系材料,为优良QTL性状的发掘和筛选奠定了材料基础,拓宽了玉米新品种选育的种质资源范围。     

基于生物信息学的玉米抗病QTL比较定位

以玉米遗传连锁图谱IBM2 2008 Neighbors为参考图谱,利用BioMercator 2.1软件,通过映射来自不同实验中的340个玉米抗病QTL,构建出玉米抗病QTL的整合图谱。采用元分析技术,在1、3、6、10号染色体上发掘5个"一致性抗病QTL"区间,图距分别为5.14cM、9.00cM、28.50cM、1.73cM和33.34cM。从MaizeGDB网站下载"一致性抗病QTL"区间内的基因和标记原始序列,采用NCBI网站在线软件BlastX通过同源比对,在5个"一致性抗病QTL"区间内初步确定8个抗病基因同源序列。借助比较基因电子定位策略,将54个水稻和44个玉米抗性基因转定于玉米IBM2 2008 Neighbors遗传连锁图谱上。本文研究结果为玉米抗病QTL精细定位和克隆奠定了基础。     

玉米丝裂病的抗性QTL定位

选用感丝裂病的玉米自交系R08与抗丝裂病的自交系Es40组配F₂群体共348个单株,构建了包含115个SSR标记的分子遗传连锁图谱,覆盖玉米基因组2 178.6 cM,平均图距为18.9 cM。采用复合区间作图法,对F_2:4家系丝裂病数据进行抗性QTL分析,共检测到12个QTL,分别位于第1、2、4、5和7染色体,贡献率为4.22%~37.95%。其中在第1、3染色体上检测到主效QTL,贡献率均大于30%,基因作用方式均为显性,其余10个QTL的作用方式多为加性或部分显性。     

玉米出籽率、籽粒深度和百粒重的QTL分析

为研究玉米出籽率、籽粒深度、百粒重的遗传机制,以豫82×沈137组配的229个F_2:3家系为试验材料,采用复合区间作图法进行QTL定位分析。在3个环境下共检测到10个QTL。其中,控制出籽率、籽粒深度、百粒重相关QTL分别为3个、3个和4个,它们的联合贡献率分别为35.5%、28.1%和39.0%。位于第1染色体上介于标记umc1335与umc2236之间控制出籽率的QTL qKR1b和位于第9染色体上介于标记bnlg1209–umc2095之间控制百粒重QTL q100-KW9b,分别解释18.9%和11.7%的表型变异,且作用方式为加性效应,分析表明这些区域可能包含调控玉米籽粒性状关键基因,对剖析玉米产量形成机制具有重要的参考价值。     

基于单片段代换系的玉米百粒重QTL分析

籽粒大小是影响玉米产量的关键因素。本研究基于59份玉米染色体单片段代换系(SSSL)纯合体,对玉米百粒重性状进行2年6个试验环境的表型鉴定,利用t测验和重叠群作图的方法对SSSL内代换片段上的百粒重效应进行了QTL分析。共检测出20个百粒重QTL,分布在玉米的8条染色体上,其中14个(70.0%)在2个以上试验环境中被重复检出,4个(20.0%)在4个以上试验环境中被重复检出,在全部6个试验环境中重复检出且基因加性效应值较大的玉米百粒重QTL是位于玉米第5染色体Bin5.05区SSR分子标记bnlg278和umc1680附近的q100kw-5-3。研究结果为玉米籽粒大小性状相关基因的进一步精细定位和克隆奠定了基础。     

利用种子性状QTL定位高油玉米蛋白质含量QTL

利用普通玉米自交系8984与高油玉米自交系GY220为亲本构建了284个F2∶3家系群体及含有185个SSR标记的玉米遗传连锁图谱。通过包含母体效应的种子性状QTL作图方法对玉米子粒蛋白质含量进行定位和效应分析,共检测到4个QTL,位于第5和第8染色体上。除qPRO8-2遗传作用方式表现为加性外,其余QTL作用方式均为部分显性。单个QTL贡献率为3.86%~5.17%,累计贡献率为18.54%。所有QTL的增效基因均来自高油亲本GY220。     

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。     

基于Meta分析构建抗玉米粗缩病的染色体单片段代换系

为发掘玉米粗缩病抗性基因,解析其遗传规律。借助IBM2 2008 Neighbors遗传图谱,整合已报道的92个抗玉米粗缩病QTLs(Quantitative trait locus),通过Meta分析获得24个"一致性"抗病区间。利用生物信息分析,确定抗病区段的物理位置信息,在相关区段对郑58和齐319的全基因组重测序序列分析鉴定出7 142个InDel位点,其中546个含有InDel位点的序列可用于引物开发,进一步通过试验筛选出在郑58和齐319第2,4,5,6,7,8,10号染色体上的多态InDel位点158个。以抗粗缩病玉米自交系齐319为供体亲本,感病优良自交系郑58为受体亲本,利用上述InDel标记辅助选择构建整套染色体单片段代换系材料。为抗玉米粗缩病QTL位点的精细定位提供了可能,也为抗病育种提供新的种质资源。     

QTL analysis for eating quality-related traits in an F2:3 population derived from waxy corn × sweet corn cross

In order to identify quantitative trait loci (QTL) for the eating quality of waxy corn and sweet corn (Zea mays L.), QTL analysis was conducted on an F₂ population derived from a cross between a waxy corn inbred line and a sweet corn inbred line. Ten QTLs for pericarp thickness (PER), amylose content (AMY), dextrose content (DEX) and sucrose content (SUC) were found in the 158 F₂ families. Among them, four QTLs, qAMY4 (10.43%), qAMY9 (19.33%), qDEX4 (21.31%) and qSUC4 (30.71%), may be considered as major QTLs. Three of these, qAMY4, qDEX4 and qSUC4, were found to be located within a region flanked by two adjacent SSR markers on chromosome 4 (umc1088 and bnlg1265), making this SSR marker pair a useful selection tool for screening the eating quality traits of AMY, DEX and SUC. The QTL for amylose content was found to be located between markers phi027 and umc1634, raising the possibility of its identity being the Wx1 gene, which encodes a granule-bound amylose synthase. The new QTLs identified by the present study could serve as useful molecular markers for selecting important eating quality traits in subsequent waxy corn breeding studies.     

Identification of QTL for stalk sugar-related traits in a population of recombinant inbred lines of maize

Increasing sugar content in silage maize stalk improves its forage quality and palatability. The genetic mapping and characterization of quantitative trait loci (QTLs) is considered a valuable tool for trait enhancement, yet little information on QTL for stalk sugar content in maize has been reported. To this end, we investigated QTLs associated with stalk sugar traits including Brix, plant height (PHT), three ear leaves area (TELA), and days to silking (DTS) in two environments using a population of 202 recombinant inbred lines from a cross between YXD053, which has a high stalk sugar content, and Y6-1, which has a low stalk sugar content. A genetic map with 180 SSR and 10 AFLP markers was constructed, which spanned 1,648.6 cM of the maize genome with an average marker distance of 8.68 cM, and QTLs were detected using composite interval mapping. Seven QTLs controlling Brix were mapped on chromosomes 1, 2, 6 and 9 in the combined environments. These QTLs could explain 2.69–13.08 % of the phenotypic variance. One major QTL for Brix on chromosome 2 located between the markers bnlg1909 and umc1635 explained 13.08 % of the phenotypic variance. Y6-1 also contributed QTL allele for increased Brix on chromosome 6. One major QTLs controlling PHT on chromosome 1 and TELA on chromosome 4 were also identified and accounted for 13.68 and 12.49 % of the phenotypic variance, respectively. QTL alleles for increased DTS were located on chromosomes 1 and 5 of YXD053. Significant epistatic effects were identified in four traits, but no significant QTL × environment interactions were observed. The information presented here may be valuable for stalk sugar content improvement via marker-assisted selection in silage maize breeding programs.     

Identification of QTL for acid phosphatase activity in root and rhizosphere soil of maize under low phosphorus stress

To provide theoretical and applied references for acid phosphatase (APase) activity of maize, this study was to identify quantitative trait locus (QTL) for APase activity in root and rhizosphere soil of maize under low phosphorus (p) stress. The correlation and the QTL of APase activity in root (APR) and APase activity in rhizosphere soil (APS) were studied for the F_2:3 population derived from the cross 082×Ye107 under low P stress in two sites. Analysis for each environment and joint analysis across two environments were used to identify QTL for the F_2:3 population. A significant difference in APR and APS was found between 082 (P efficient genotype) and Ye107 (P deficient genotype). A large genetic variation and transgressive segregation of the F_2:3 population were observed in Beibei and Hechuan. One stable QTL for APR was detected in different environments, which was located in the interval bnlg1350 to bnlg1449 on chromosome 3. Two stable QTLs for APS were detected, which were located in the interval umc2083 to umc1972 on chromosome 1 and in the interval umc2111 to dupssr10 on chromosome 5. The stable QTLs can be used in MAS breeding and theoretical study of maize.     

玉米幼胚愈伤组织的诱导和植株再生的QTL分析

以黄早四和Mo17为亲本组配的239个RIL群体，构建了101个SSR标记的遗传图谱，覆盖玉米基因组1 422.7 cM，标记间的平均距离为15.6 cM。以玉米幼胚为外植体、改良N6为基本培养基，对亲本及RIL群体的组培性状进行了评价。采用复合区间作图法在第2、3、5、6、8和9染色体上定位了控制出愈率、Ⅱ型愈伤组织诱导率、绿点及绿苗分化率的8个QTL，并对其遗传效应进行了分析，其基因加性效应能解释相应性状表型方差的4.78%~14.02%。     

大田环境下玉米抗旱相关性状QTL定位

干旱是世界范围内导致玉米产量损失的主要因素。为了阐明玉米抗旱性的遗传基础并定位相关的数量性状位点,利用抗旱自交系临1和敏感的湘97-7组配160个F2:3家系定位群体,于2011年在湖南省作物研究所和长沙县高桥镇,分别在大田干旱胁迫和正常水分条件下进行表型鉴定。所考察性状包括抽雄至吐丝间隔、株高、千粒重和产量,用抗旱系数来衡量抗旱性。结果表明,110个SSR标记构建连锁图,图谱总长1246.1 cM,标记间平均距离11.33 cM。抗旱相关性状定位的QTL介于8～14个,共检测到43个QTL。单个QTL解释的表型变异为6.27%～18.27%。不同水分条件下定位到的QTL大多数不相同,表明对干旱胁迫的适应存在不同机制。抗旱性相关性状定位到的QTL,除第2和10染色体外,在其它染色体上都有分布,主要集中在第1染色体1.02-03区域和1.06-07区域,以及第3染色体3.04-05区域。第1染色体标记umc2224和bnlg176区间同时检测到与株高、千粒重和产量有关的QTL簇;标记bnlg1556和umc1128区间检测到与抽雄至吐丝间隔和产量有关的QTL簇。第3染色体标记umc1773和umc1311区间同时检测到与株高、千粒重和产量有关的QTL簇。这些QTL簇可能有助于通过分子标记辅助选择的方法提高干旱地区玉米的抗旱性。     

Identification of quantitative trait loci for drought tolerance at seedling stage by screening a large number of introgression lines in maize

The maize genome hosts tremendous phenotypic and molecular diversity. Introgression lines (ILs), developed by continuous backcrossing to recurrent parents, could provide a unique genetic stock for quantitative trait locus (QTL) mapping. Using maize lines from six heterotic groups of different ecological zones, we developed >500 BC₂F₂ IL sets by crossing 11 inbred lines (as recurrent parents) with >200 local maize inbred lines (as donor parents). Of them, 34 IL sets were selected as a subset for drought tolerance screening and a total of 417 ILs survived under severe water stress at seedling stage. One set of 32 surviving ILs, derived from Chang7-2/DHuang212, was used for QTL mapping with simple sequence repeat markers covering the whole genome, with seven QTL detected. Furthermore, investigating all surviving ILs, we identified two common regions in bin 3.04, corresponding to marker intervals bnlg1904–umc1772 and umc1223–bnlg1957, respectively, which shared high genetic variation in three IL sets. Our results indicated that selective genotyping can be used to identify genetic loci for complex traits. The ILs, highly selected for drought tolerance in this study, provide a unique set of materials for both genomic studies and development of enhanced germplasm resources.     

玉米光周期敏感相关性状发育动态QTL定位

玉米是短日照作物,大多数热带种质对光周期非常敏感。光周期敏感性限制了温、热地区间的种质交流。研究玉米光周期敏感性的分子机理,有利于玉米种质的扩增、改良、创新,提高玉米品种对不同光周期变化的适应性。本研究以对光周期钝感的温带自交系黄早四和对光周期敏感的热带自交系CML288为亲本配置的组合衍生的一套207个重组自交系为材料,在长日照环境条件下对不同发育时期的叶片数、株(苗)高变化进行QTL分析。结果表明,双亲间的最终可见叶片数和株高差异很大;发育初期CML288的叶片数和苗高都低于黄早四,而发育后期CML288的叶片数和株高都明显高于黄早四;测定各时期F₇重组自交系间也存在显著差异。利用包含237个SSR标记、图谱总长度1 753.6 cM、平均图距7.40 cM的遗传连锁图谱,采用复合区间作图法,分别检测到控制叶片数和株(苗)高发育的QTL 11个和20个。但是,没有一个条件QTL 能在测定的几个时期都有效应。在长日照条件下,控制叶片数与株(苗)高的非条件与条件QTL主要集中在第1、9和10染色体上,特别是在第10染色体的标记umc1873附近均检测到了影响这两个性状的QTL,且在不同的发育时期单个条件和非条件QTL所解释的表型变异分别为4.34%~25.74%和10.02%~22.57%,表明这一区域可能包含光周期敏感性关键基因。