山西省主推玉米杂交种纯度鉴定SSR核心引物的筛选

选用10对SSR核心引物对18份山西省玉米杂交种进行DNA指纹分析,并综合考虑多态性和杂合率,认为umc1590和umc1196这2对引物可鉴定17个品种纯度,phi402893,phi102228,phi233376,p-phi072,umc1066和phi022也可作为纯度鉴定的引物。建议采用umc1590,phi402893,phi233376,phi022这4对引物对18个杂交种纯度进行检测。     

玉米SSR标记的多重PCR试验分析

以玉米自交系4011为试验材料,选用玉米SSR核心引物20对,利用单重SSR分子标记技术体系,采用逐步递加引物对的方法,尝试了二重、三重、四重、五重、六重PCR试验.结果筛选出了6个适宜六重PCR扩增的SSR标记组合:phi053/umc2084/umc1196/bnlg1154/mmc0151/bnlg1025、umc1231/umc1196/phi053/umc1222/mmc0151/bnlg1025、umc1231/umc1196/phi053/umc1222/bnlg240/bnlg1025、umc1231/umc1196/phi053/umc1222/1154/bnlg249、bnlg1451/umc1196/phi053/umc1019/phi072/bnlg1025、bnlg1451/umc1196/phi053/umc1545/bnlg2291/bnlg1025.这些标记组合的PCR扩增和PAGE电泳检测良好,从而可提高SSR标记检测的效率,并且降低试验成本.     

opaque-2 基因导入甜糯玉米表型分析及导入系群体初步创制

[目的]甜糯玉米是华南地区重要的经济作物,但籽粒中赖氨酸含量低,探索研究opaque2(o2)导入对甜糯玉米籽粒表型及赖氨酸积累的影响.[方法]通过常规杂交方法在甜糯玉米中引入o2基因,利用o2基因内紧密连锁的SSR共显性标记检测供体和甜糯玉米受体自交系的多态性,利用其特异性和共显性检测回交自交后代.[结果]投射光下供体亲本HP3籽粒表现不透光,受体材料透光性良好;杂交穗当代表现不同基因型籽粒分离,出现硬质、皱缩等表型,回交子代籽粒透光性明显降低,可能与o2基因存在有关.对o2基因的3对SSR标记umc1066、phi112和phi057筛选,只有标记phi057在供体和受体自交系中呈现良好的多态性,进一步研究发现,在回交和自交后代中能够检测到供体的o2突变基因成功导入甜糯玉米受体自交系.[结论]通过子代籽粒表型及SSR标记phi057筛选鉴定,初步创制了Opaque-2甜/糯玉米早代群体系;SSR标记phi057筛选出F1与BC1代o2基因植株,说明phi057标记可有效区分含o2基因植株.     

太空诱变玉米核不育基因的微卫星标记

利用随机交配多代的郑58可育株5280(Ms)与不育株5280(ms)杂交、昌7-2不育株8057(ms)与正常可育自交系黄C杂交,分别构建两个F2定位群体.在田间育性鉴定的128株和146株中,可育株与不育株分离分别为93株完全可育、35株完全不育和114株完全可育、32株完全不育.适合性测验表明,均符合3:1孟德尔遗传分离比例.运用SSR分子标记技术和BSA分组方法.分别选用遍布在玉米10条染色体上的418对和386对SSR标记对两个群体进行多态性筛选,结果对应有17对和36对引物出现多态性.对两个F,定位群体进行基因型和连锁分析结果表明,郑58和昌7-2太空诱变玉米细胞核雄性不育基因分别位于第1和第4染色体上,微卫星标记phi 427913和umc 1160与郑58核不育基因连锁,遗传距离分别为23.8和19.4 cM;phi 006和umc 1109与昌7-2核不育基因连锁,遗传距离分别为18.0和26.3 cM.     

优良玉米杂交种金玉818指纹图谱构建及其纯度鉴定

为构建优良杂交玉米新品种金玉818的指纹图谱,筛选适用于该品种真实性和纯度鉴定的SSR引物,利用SSR分子标记技术构建指纹图谱,筛选引物对金玉818种子进行纯度鉴定,并对鉴定结果进行验证。结果表明:在40对标准SSR引物中筛选出25对多态性好、清晰稳定的引物构建金玉818指纹图谱,筛选出15对适用于该品种真实性和纯度鉴定的引物,其中最佳引物是bnlg16lk8、umc1125y3、umc1429y7、phi053k2、umc1147y4和bnlg2305k4;利用phi053k2、bnlg16lk8和umc1429y7鉴定金玉818种子的纯度分别为92.5%、92%和93.5%,引物间鉴定结果吻合度高,结果可靠。     

SSR标记鉴定玉米品种真实性及纯度的研究

[目的]探讨杂交玉米及其亲本种子真实性和纯度的分子检测判读标准。[方法]以16个玉米杂交种及其双亲和202个骨干自交系为引物筛选材料,分别构建3个杂交种和3个自交系人工群体的验证试验材料,用CTAB法提取幼苗叶片DNA,选137对SSR引物进行SSR扩增及产物检测,用于玉米品种的真实性鉴定和纯度检测,并与同批移栽至大田的种植鉴定结果相比较。[结果]从137对SSR引物中筛选出了多态性信息含量（PIC值）高、扩增条带清晰和重复性好的20对一级和20对二级核心引物,用一级核心引物对杂交种人工群体进行真实性鉴定,根据一级核心引物的扩增结果,从中选取3对（phi080、umc1196、umc2084）表现较好的引物用于杂交种人工群体的纯度检测。ssR分子检测结果与田间种植鉴定结果具有高度一致性。[结论]利用SSR标记技术鉴定玉米品种真实性和纯度的方法是可行的。     

玉米杂交种子纯度的SSR检测与田间鉴定的比较研究

为发展分子标记技术在玉米杂交种子纯度鉴定上的应用,文章对吉林省2005年玉米省级区试的30个品种进行了SSR分子标记的纯度分析,筛选出11时多态性好的引物,并与田间纯度鉴定结果进行了比较.通过相关分析和差异显著性分析表明:SSR鉴定的(平均)纯度与田间纯度鉴定结果较为一致,相关系数达到0.798,而且两者无显著差异,因此,SSR分子标记可以用于玉米田间纯度的鉴定;引物umc1069和引物umc2373所标记的结果能较好的代表田间纯度鉴定的结果.     

利用SSR技术鉴定苏玉20的纯度

苏玉20是江苏省农业科学院2004年选育成功的高产优质玉米杂交种.以苏玉20玉米杂交种及其亲本为材料,从80对玉米SSR引物中筛选出在苏玉20双亲之间多态性明显、重复性较好的4对引物bnlg1306、phi065、bnlg2291和umc1590用于苏玉20纯度鉴定.这4个SSR标记特异性好,且人为掺杂与田间鉴定均与利用SSR标记鉴定结果高度一致.尤其是,这些引物同时可以用于郑单958的纯度鉴定并可以区分苏玉20和郑单958杂交种.因此,该方法具有准确可靠、实用性强等优点,为生产上杂交种苏玉20纯度的快捷有效鉴定提供了科学依据.     

利用SSR标记鉴定玉米杂交种南校18号种子纯度

以南校18号玉米杂交种及其亲本自交系为材料,对胚芽和干种子的DNA进行提取,对24对SSR引物进行筛选,获得适合该杂交种纯度鉴定的SSR引物。结果表明,在快速检测中,从干种子胚提取DNA更加便捷、快速,无DNA降解,可用于PCR扩增反应;24对SSR引物中,有3对特异性引物6mmc0241、umc2025、phi323152可用于南校18号杂交种和亲本自交系的纯度鉴定。因此,应用SSR标记结合单籽粒DNA快速提取方法,可以快捷、准确地鉴定玉米杂交种种子纯度。     

SSR标记在糯玉米品种鉴定上的应用

采用NY/T 1432—2014《玉米品种鉴定技术规程SSR标记法》中的20对简单重复序列(SSR)引物对市场上抽检的苏玉糯11、苏玉糯901、苏玉糯13、苏玉糯14、苏玉糯639等5个糯玉米品种进行真实性鉴定。结果表明,每个抽检品种与标准品种至少有6个差异位点,这5个品种均与相应的标准品种不同;此外,umc1545y2、umc1335y5、umc2105k3、umc1705w1、umc1125y3、umc2007y4、phi053k2、bnlg2305k4等8对引物多态性较好,在今后糯玉米品种鉴定中可优先采用。     

玉米矮秆主效QTL qph1-4的精细定位

利用1份在玉米自交系87-1的遗传背景上综3的染色体单片段代换系(Single segment substitution line,SSSL) SSSL-Y7为试验材料,3年3点的株高表现型鉴定表明,SSSL-Y7的株高均显著矮于受体亲本87-1,且加性效应百分率均在-10％以下,推测在该SSSL内的位于第1染色体长臂上SSR分子标记umcl122附近的目标代换片段上存在可以使玉米株高致矮的主效QTL.在该SSSL与87-1杂交构建的F2分离群体中,高秆与矮秆的株数符合3∶1的分离比例,推测其矮秆表现型由1对隐性基因控制,将该基因命名为qph1-4.qph1-4基因来源于供体自交系综3,位于Bin 1.07区域,在SSR标记MPH147和umc2396之间,距两标记的遗传距离分别为1.5和0.3 cM.与qph1-4基因连锁的SSR标记还有MPH164,umc1122,MPH162,MPH9,qph1-4与之间的遗传距离分别是2.2,2.0,2.0和2.5 cM,这些SSR标记与qph1-4基因在染色体上的排列顺序为MPH164-umc1122-MPH162-MPH147-qph1-4-umc2396-MPH9.     

簇毛麦易位系V9125-2抗条锈基因YrWV的遗传分析和SSR分子标记

[目的]对高抗条锈病的簇毛麦易位系V9125-2进行研究,明确其抗病性遗传特点,并对其抗条锈病基因定位,为选育优质抗源材料提供依据.[方法]采用中国当前流行的7个条锈菌生理小种CYR29、CYR30、CYR31、CYR32、CYR33以及Su11-4、Su11-11对簇毛麦易位系V9125-2和铭贤169的杂交后代进行...     

玉米白化突变体As-81647的鉴定及基因定位

白化突变体As-81647是在自交系81647的自交后代中发现的自然突变体。光合色素含量测定表明,白化突变体叶绿素a、叶绿素b、叶绿素总量分别为正常植株的0．6％、5．8％和1．6％,推测光合色素含量的减少导致突变体光合作用强度减弱,无法利用光能进行营养生长,进而造成突变体产生白化表型,三叶期左右萎蔫死亡。组织细胞染色实验证实H2O2的积累可能是造成突变体细胞死亡的直接原因之一。本研究构建了As-81647杂合突变体与蒙自2的杂交F2分离群体,遗传分析表明该白化性状由一对隐性核基因控制,暂时命名为As-81647（Albino seeding一81647）。利用已公布的SSR分子标记和本实验室开发的分子标记,将该基因定位在玉米第3染色体umc1052和ume1641之间,遗传距离分别为0．7cM和2．8cM且与分子标记as47共分离。     

Evaluation and Quantitative Inheritance of Several Drought-Relative Traits in Maize

The authors evaluated 57 parental inbred lines of maize hybrids disseminated in Southwest China for drought tolerance under drought-stressed and well-watered conditions. Multiple regression analyses between drought tolerant coefficients of the grain yield per plant and 15 morphological and physiological traits measured from a subset of 12 selected lines, identified traits 1 and 5, which were important for drought tolerance, at the seedling and reproductive stages respectively. Gene effects, combining abilities, and heritabilities of these traits were estimated using generation mean and diallel cross methods. Dominance effect was more important than additive effect for the plant height, anthesis-silking interval （ASI）, root weight, and the grain yield per plant, whereas, they were about equal for the leaf emergence rate. The variances of special combining ability （SCA） were about double that of the general combining ability （GCA） for plant height, ASI and grain yield per plant, although they were about equal for leaf emergence rate and root weight. Narrow sense heritabilities of the five traits for the reproductive stage were not high （12.8-29.6%）, although broad sense heritabilities for plant height, ASI, and grain yield were as high as 70-85%. A segregating population consisting of 183 F2 plants from the cross N87-1 （drought tolerant） × 9526 （susceptible）, was genotyped at 103 SSR loci and the F2：4 families were evaluated under two water regimes. Twelve quantitative trait loci （QTLs） （two for plant height, five for ASI, four for root biomass, and one for grain yield） were identified, most of which had overdominant gene action. Some chromosomal regions, such as those linked to markers umcl051 （bin 4.08）, umc2881 （bin 4.03）, and phi034 （bin 7.02）, had overlapping QTLs.     

玉米C型胞质不育恢复主基因SSR标记

 通过对恢复系凤可 １、Ａ６１９与 Ｃｍｓ－Ｃ２３７、Ｃｍｓ－ＣＭｏ１７组配的 Ｆ２和 ＢＣ１分离群体的研究结果表明：凤可 １有两对重叠恢复基因 Ｒｆ４和Ｒｆ５。用微卫星标记（ＳＳＲ）将凤可１中的恢复主基因Ｒｆ５定位在第 ５染色体长臂上，与引物 ｂｎｌｇ１７１１、ｂｎｌｇ１３４６和 ｐｈｉ０５８紧密连锁，距 ３个引物的遗传距离分别为 ７．５１ｃＭ、１．６８ｃＭ、９．８７ｃＭ；恢复主基因 Ｒｆ４与第８染色体短臂上的引物ｂｎｌｇ２３０７连锁。     

QTL Mapping of Hard Seededness in Wild Soybean Using BSA Method

【Objective】 Hard seededness of wild soybean is an important effector that limits the utilization of wild resources in soybean genetic improvement. Bulked segregant analysis (BSA) was employed to identify major quantitative trait loci (QTLs) related with hard seededness in soybean, which laid a foundation for effective utilization of wild soybean germplasm in cultivated soybean improvement. 【Method】 F₂ and F₇ segregation populations were constructed from a cross between cultivated soybean Zhonghuang39 and wild soybean NY27-38. Uniformly sized seeds were selected from each line, and 30 seeds were soaked in a petri dish with 30 mL distilled water for 4 hours at 25℃. The assay was replicated 3 times. The number of permeable and impermeable seeds were counted. In F₂ population, the first DNA pool was constructed from 22 individuals with permeable seeds (imbibition rate ＞90%), and second DNA pool was constructed from 16 individuals with impermeable seeds (imbibition rate ＜10%). In F₇ population, 20 lines with permeable seeds (100% imbibition) and 20 lines with impermeable seeds (no imbibition) were used to construct two DNA pools, respectively. To detect genomic regions associated with hard seededness, these DNA bulks were genotyped with 259 polymorphic SSR markers to identify markers linked to QTL. A linkage map was constructed with 192 SSR markers, QTLs related with hard seededness were identified by composite interval mapping in F₇ segregation population. 【Result】 Out of 259 SSR loci polymorphic between Zhonghuang39 and NY27-38, 10 and eight polymorphic SSR markers between the permeable and impermeable pools were detected in 16.3 Mb interval on chromosome 2 and 23.4 Mb interval on chromosome 6, respectively, in F₂ population. The QTL region (276.0 kb) located between Satt274 and Sat_198 on chromosome 2 contained previously cloned gene GmHs1-1, the QTL explained 17.2% of the total genetic variation. The other QTL was mapped on chromosome 6 flanked by BARCSOYSSR_06_0993 and BARCSOYSSR_06_1068, accounting for 17.8% of the total genetic variation. In F₇ population, eleven, nine and four SSR polymorphic markers between the permeable and impermeable pools were detected in 27.4 Mb interval on chromosome 2, 27.8 Mb interval on chromosome 6, 18.2 Mb interval on chromosome 3, respectively. A linkage map of 192 SSR markers and covering 2 390.2 cM was constructed through composite interval mapping in F₇ population. Three QTLs related with hard seededness were detected. The QTL on chromosome 2 located between Satt274 and Sat_198, explained 23.3% of the total genetic variation; the QTL on chromosome 6 flanked by Sat_402 and Satt557, explained 20.4% of the total genetic variation; the QTL on chromosome 3 flanked by Sat_266 and Sat_236 accounted for 4.9% of the total genetic variation. 【Conclusion】 In this study, three QTLs related to soybean hard seededness were identified by both BSA and traditional linkage mapping, indicating that BSA is an effective strategy for identifying QTLs in soybean.     

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。