Perennial aneuploidy as a potential material for gene introgression between maize and Zea perennis

Hybridization,which allows for gene flow between crops,is difficult between maize and Zea perennis.In this study,we aim to initiate and study gene flow between maize and Z.perennis via a special aneuploid plant(MDT) derived from an interspecific hybrid of the two species.The chromosome constitution and morphological characters of MDT as well as certain backcross progenies were examined.Results from genomic in situ hybridization(GISH) indicate that aneuploid MDT consisted of nine maize chromosomes and 30 Z.perennis chromosomes.The backcross progenies of MDTxmaize displayed significant diversity of vegetative and ear morphology;several unusual plants with specific chromosome constitution were founded in its progenies.Some special perennial progeny with several maize chromosomes were obtained by backcrossing MDT with Z.perennis,and the first whole chromosome introgression from maize to Z.perennis was detected in this study.With this novel material and method,a number of maize-tetraploid teosinte addition or substitution lines can be generated for further study,which has great significance to maize and Z.perennis genetic research,especially for promoting introgression and transferring desirable traits.     

玉米重组自交系苗期耐盐相关性状QTL的初步定位

[目的]对玉米重组自交系苗期耐盐相关性状进行QTL定位。[方法]以玉米黄早四与Mo17杂交的RIL-F7代171份材料为作图群体,构建其玉米的分子标记遗传图谱,并在此基础上开展对玉米耐盐相差性状的QTL分析。[结果]构建了一张包含81个SSR位点,覆盖了玉米基因组10条染色体,共1 428.3 cM,标记间平均间距为17.63 cM的分子遗传连锁图谱;共检测到6个QTL,分别位于第1、5、6号染色体上。[结论]该研究对深入认识玉米耐盐遗传机理,了解控制玉米耐盐性的基因数目及其在染色体上的位置以及对耐盐性的遗传效应,进行玉米耐盐性育种的分子标记辅助选择和基因克隆均具有极其重要意义。     

Study on Haploid Inducing and Its Meiotic Abnormality in Maize

The haploid-inducing line Stock 6 was used to produce haploid maize and expected to obtain maize haploid plants successfully. The detailed meiotic studies on selected haploid maize （n = x = 10） were conducted. Cytogenetic analysis revealed a high frequency of meiotic abnormality occurred in both meiosis Ⅰ and meiosis Ⅱ. During the prophase Ⅰ, univalents were common configurations, and there were bivalents or trivalents in some pollen mother cells, however, a few cells containing five bivalents were also observed. After prophase Ⅰ, chromosomes did not congregate in a single metaphase plate but they were scattered in the cytoplasm. At anaphase Ⅰ, the chromosome distribution was highly irregular with almost all possible combinations. In some cells, chromosomes were grouped into the three or four masses and several spindles appeared. At the tetrad stage of meiosis Ⅱ, eytokinesis splitting abnormality occurred, and a variety of diad, triad, tetrad, pentad, hexad, as well as decury microspores were easily observed. As a consequence of abnormalities of the two meiotic stages, various microspores and the pollen were almost completely sterile. grains with different size were formed, and its pollen grains     

Discovery of transposable element activity among progeny of tissue culture--derived maize plants

Tissue culture-derived plants of many species have often been observed to possess both genetic and cytogenetic abnormalities. A high frequency of structurally altered chromosomes in maize (Zea mays L.) plants regenerated from tissue culture led to the prediction that newly activated transposable elements could be detected in regenerated plants. Testcrosses of 1200 progeny from 301 regenerated maize plants confirmed that ten regenerated plants from two independent embryo cell lines contained an active Actransposable element. No active Ac elements were present in the explant sources. Recovery of transposable element activity in regenerated plants indicates that some tissue culture-derived genetic variability may be the result of insertion or excision of transposable elements, or both.     

化学杀雄剂SQ-1和阿拉伯葡聚糖蛋白对小麦品种间杂交及远缘杂交成胚率的影响

【目的】探究化学杀雄剂SQ-1对小麦品种间、小麦与近缘植物间、小麦与远缘植物间杂交成胚率的影响,以及阿拉伯葡聚糖蛋白对小麦与玉米杂交成胚率和得苗率的影响。研究结果对于合理选用小麦杂交方式,提高小麦杂交结实率和利用玉米诱导小麦单倍体植株的效率具有重要意义。【方法】通过在小麦拔节期喷施化学杀雄剂SQ-1,开花期分别授以小麦花粉和远缘植物(黑麦、玉米)花粉,并在小麦授玉米花粉后的处理液中加入不同浓度的阿拉伯葡聚糖蛋白(arabinogalactan proteins,AGP),对小麦与玉米杂交后产生的幼胚进行离体拯救培养,统计授粉小花数、接种幼胚数、膨大颖果数、结实粒数、萌发单倍体幼胚数和单倍体植株数,计算结实率、颖果膨大率、成胚率、萌发率和成苗率并对所得数据进行差异显著性分析,结合细胞学观察结果,研究SQ-1对小麦品种间杂交及远缘杂交结实性的影响,以及AGP对小麦单倍体胚诱导率的影响。【结果】在不同小麦品种间杂交中SQ-1处理结实率19.8%—83.3%,人工去雄的结实率为69.4%—93.0%,SQ-1对不同品种的影响不同,Fielder对SQ-1的反应比较敏感;在中国春与兰州黑麦杂交中,SQ-1处理的结实率为65.5%,人工去雄处理的结实率为78.8%,两种处理方式产生的F1杂种的染色体数均为28条;在不同小麦品种与玉米品种郑单58杂交中,SQ-1处理小麦单倍体胚的成胚率为1.11%—1.41%,人工去雄小麦单倍体胚的成胚率为2.38%—14.29%;在小麦与玉米杂交后的处理液中添加0.5—2.0 g·L-1 AGP一定程度地提高了小麦单倍体胚获得率和成苗率。另外,在玉米花粉诱导的单倍体胚离体培养过程中,发现13.07%的胚发育出了2—6株苗;显微镜观察发现,玉米花粉诱导后18 d左右小麦单倍体胚上出现了多个突起,这些突起在离体培养条件下进一步发育为形态健全的小植株,其染色体数目均为21条。【结论】化学杀雄剂SQ-1减低了小麦品种间杂交及小麦与黑麦、玉米间杂交的成胚率,AGP提高了小麦与玉米间杂交单倍体成胚率和成苗率。     

玉米Glyco-hydro-16糖苷酶家族全基因组的鉴定及其遗传分化

【目的】全基因组水平鉴定玉米Glyco-hydro-16家族,分析该家族基因在不同组织中的表达模式以及在不同玉米杂种优势群中的遗传分化。【方法】根据Glyco-hydro-16家族相对保守的序列及结构域,构建Glyco-hydro-16家族的隐马尔科夫模型文件（Glyco-hydro-16.hmm）,利用hmmersearch程序在玉米全基因组中进行比对,获得玉米中含有该家族保守结构域的所有序列。通过Blast2GO进行功能注释,利用蛋白质序列构建该家族的系统发育进化树。使用玉米自交系B73不同组织及不同发育时期的RNA-seq数据库分析该家族基因的表达模式。根据该家族基因在染色体上的位置筛选SNP标记,计算其在不同玉米杂种优势群间的群间遗传分化系数（genetic differentiation coefficient,Fst）,分析其遗传分化。【结果】根据该家族相对保守的序列及结构域,在全基因组水平共鉴定出34个玉米Glyco-hydro-16家族成员,注释表明所有基因都是木葡聚糖转移酶/水解酶基因,3个保守性较高的Motif区段存在于该家族所有成员中。通过系统发育关系和序列相似性将该家族分为8个亚家族,每个亚家族有2-8个基因,分布在除第3和第6染色体外的其他8条染色体上,在第2、第5及第10染色体上成簇分布。该家族在禾本科作物中同源性较高,与拟南芥分属不同的分支,但只有3个玉米成员（AC210669.3、GRMZM2G413006和GRMZM2G166944）被划分到禾本科分支中,其他玉米成员被划分到单独的分支中。通过表达谱分析表明该家族成员在玉米中均有表达,但在不同组织中的表达水平有差异。为解析该家族基因在不同玉米种质资源中等位基因的变异,根据玉米Glyco-hydro-16家族基因在染色体上的位置筛选SNP标记,计算其在玉米杂种优势群SS及NSS间的群间遗传分化系数。结果显示,共有10个该家族基因所处位点的Fst值高于阈值0.15,达到高度分化水平,分别位于第1、第2、第4、第5、第7以及第9染色体上。其中,位于第2染色体上的GRMZM2G091118相应位点的Fst值为0.52,表明该位点在SS群和NSS群间的群间遗传分化度极大。【结论】通过全基因组扫描在玉米中鉴定出34个Glyco-hydro-16家族成员,均为木葡聚糖转移酶/水解酶基因,在不同组织中,其表达模式不同,可能参与不同生理发育过程。部分该家族成员所处位点在玉米杂种优势群SS和NSS间的等位基因分化极大。     

Breeding for Parents of Hybrid Rice with Maize pepc Gene

The results of the investigation on transgenic rice with maize C4-specific phosphoenolpyruvate carboxylase (pepc) gene showed that the transgenic rice plants with the maize pepc gene expressed at high level and the maize PEPC expression was inherited in the progenies in a Mendelian manner. The transgenic plants had PEPC activity of more than 10-fold higher than untransformed plants. As compared with untransformed plants, the panicle per plant, spikelet per panicle, 1000-grain weight and grain-weight per plant for transgenic plants increased by 14.9 %, 5.7%, 1.3 % and 13.9 %, respectively. By crossing the maize pepc gene was incorporated into the parents of hybrid rice, which were the photo-sensitive genic male sterile (PGMS)lines of two-line hybrid rice such as Peiai64s, 7001s, 2302s, 2304s and 2306s-1, and the BT type of cytoplasmic male sterile (CMS) line of three-line hybrid rice such as Shuangjiu A, and restorer lines 5129, Wanjing97 in the spring of 1998. The following progresses were made: (1) The inheritance of the high-level expression of the maize PEPC was stable in different genetic background of rice; (2) PEPC activity of F1 hybrid was the mean of the two parents. Its saturated photosynthetic rate (Pn) rose to 50 % higher than that of the receptor parent. These results demonstrated that it is possible to increase the vigor of the rice plant by transgenic approach with maize pepc gene; (3) The activity of PEPC in leaf could be considered as the major physiological index because the correlation coefficient between PEPC activity and Pn was 0.6470 * *; (4) We have developed three rice lines with maize pepc gene; (5) The selection method of high photosynthetic efficiency rice has been established, which includes soaking seeds into solution of hygromycin phosphotransferase to germinate, tracing the pepc gene by PCR analysis, evaluating the performance of the rice plants in the field and examining PEPC activities and Pn of rice plants with maize pepc gene.     

不同密度下玉米DH群体果穗性状的QTL定位分析

试验利用由79个DH系组成的‘农单5号’DH群体,分别在6万株/hm2和13.5万株/hm22个密度、2次重复条件下,调查了玉米穗长、秃尖长、穗粗、穗行数、行粒数、粒长、粒宽和粒厚等穗部性状,利用在双亲间存在差异且均匀分布于玉米10条染色体上的160对SSR标记对该群体进行遗传作图,并采用复合区间作图法对上述性状进行QTL分析。结果表明:在2个密度条件下共检测出30个QTL位点,单个QTL所解释的表型变异在3.26%~31.59%之间,QTL与环境之间存在复杂的互作关系。在2个密度水平下分别检测到的qEL2a(umc1165-umc1261)与qEL2b(umc1165-umc1261)可能是同一位点,qRN4a(umc2135-bnlg292a)、qRN4b(bn-lg292a-bnlg292b)和qED4(bnlg292a-bnlg292b)也很可能是同一位点。在4.08 bin区间存在着控制穗粗或穗行数的重要基因,而该基因位于umc2135-bnlg292b标记区间。     

玉米单倍体胚芽鞘节组织培养特性研究

【目的】研究玉米单倍体胚芽鞘节组织培养特性,为单倍体加倍提供一条新的途径。【方法】以Reid群、黄早四群和温热Ⅰ群群内杂交20个组合的单倍体胚芽鞘节为外植体,分析愈伤组织的诱导率和分化率,并对再生植株根尖染色体数目进行观察和花粉育性分析;同时使用SSR分子标记,分析再生植株的基因型。【结果】Reid群和黄早四群的单倍体芽鞘胚节愈伤组织的诱导率比温热Ⅰ群高,达极显著水平;共获得15株单倍体植株,根尖染色体数目为10;I-KI染色发现,散粉的花药,其花粉为部分可育;15株单倍体植株的遗传稳定,未见变异。【结论】Reid群和黄早四群的单倍体胚芽鞘节组培特性较好,温热Ⅰ群较差;组织培养产生的单倍体植株遗传型均来自双亲的重组类型。建立了玉米单倍体胚芽鞘节组织培养体系。     

ZmDREB2.7正调控玉米耐旱性的功能分析

为评价 ZmDREB2.7优异等位基因型对玉米耐旱性的作用，选择携带优异等位基因型的玉米自交系CIMBL70、CIMBL92做父本及敏感基因型的玉米自交系SHEN5003做母本，用于构建近等基因系群体。结果表明，NIL- ZmDREB2.7CIMBL70和NIL- ZmDREB2.7CIMBL92恢复率分别为 94.96%和95.11%；苗期存活率分别是81.34%和83.33%，显著高于轮回亲本（SHEN5003，37.3%）。为更加明确 ZmDREB2.7的耐旱效应，构建了 ZmDREB2.7pro: ZmDREB2.7转基因玉米植株。转基因玉米材料 ZmDREB2.7pro: ZmDREB2.7-1和 ZmDREB2.7pro: ZmDREB2.7-10相对表达量更高，苗期存活率分别是68.05%和 70.83%，显著高于野生型（A188，58.33%）。在新疆大田干旱条件下， ZmDREB2.7pro: ZmDREB2.7-1和 ZmDREB2.7pro: ZmDREB2.7-10两个株系均比野生型小区少减产20%以上。因此，研究结果可能为玉米耐旱新品种的培育提供有价值基因资源和可行的实践方案。     

介导玉米DNA的水稻高秆变异株AFLP分析

用AFLP分子标记技术,对花粉管介导玉米总DNA获得的2个高秆水稻变异植株C(HN28)和D(HN31)进行分析,揭示水稻变异植株和亲本间在DNA水平的差异,为进一步研究变异机制提供数据。首先对64对AFLP选扩引物进行筛选,优选出11对选扩引物,它们均能够在2个变异植株中扩增出较多清晰的DNA差异带。对变异植株和对照中的DNA图谱进行分析,结果显示,变异植株C、D、受体水稻B与阳性对照玉米A之间的DNA图谱相似率分别为38.30%、38.58%和32.99%,水稻变异植株C、D和受体水稻B的DNA图谱相似率分别为90.04%和85.84%,说明变异植株与受体水稻基因组DNA存在显著差异,变异植株与阳性对照玉米基因组DNA相似率明显高于受体水稻。变异植株C检测到12条与对照玉米一致的目的带,变异植株D检测到11条与对照玉米一致的目的带。说明变异植株C、D基因组中可能存在有玉米DNA片段,变异性状的差异可能是介导玉米DNA引起。     

Comparative QTL Mapping of Resistance to Gray Leaf Spot in Maize Based on Bioinformatics

The integration QTL map for gray leaf spot resistance in maize was constructed by compiling a total of 57 QTLs available with genetic map IBM2 2005 neighbors as reference. Twenty-six “real QTLs” and seven consensus QTLs were identified by refining these 57 QTLs using overview and meta-analysis approaches. Seven consensus QTLs were found on chromosomes 1.06, 2.06, 3.04, 4.06, 4.08, 5.03, and 8.06, and the map coordinates were 552.53, 425.72, 279.20, 368.97, 583.21, 308.68 and 446.14 cM, respectively. Using a synteny conservation approach based on comparative mapping between the maize genetic map and rice physical map, a total of 69 rice and maize resistance genes collected from websites Gramene and MaizeGDB were projected onto the maize genetic map IBM2 2005 neighbors, and 2 (Rgene32, ht1), 4 (Rgene5, rp3, scmv2, wsm2), and 4 (ht2, Rgene6, Rgene8 and Rgene7) positional candidate genes were found in three consensus QTLs on chromosomes 2.06, 3.04, and 8.06, respectively. The results suggested that the combination of meta-analysis of gray leaf spot in maize and sequence homologous comparison between maize and rice could be an efficient strategy for identifying major QTLs and corresponding candidate genes for the gray leaf spot.     

低磷条件下玉米‖蚕豆间作作物根系间距对玉米磷吸收的影响

为探究不同根系间距下禾本科作物玉米(生长和磷吸收)受相邻作物玉米或蚕豆的影响，本研究以玉米为目标作物，通过圆柱形根箱培养试验，设置单株玉米、单作玉米、玉米‖蚕豆间作并改变相邻作物根系间距，探究低磷条件下，不同根系间距的相邻作物对目标玉米的生长、磷吸收和根际过程的影响。结果表明:1)与单株玉米相比，目标作物玉米与相邻作物玉米和蚕豆之间均以竞争为主，当作物根系间距较近时，单作玉米的生物量、磷含量分别下降21%、33%，间作玉米分别下降34%、31%；当作物根系间距较远时，单作玉米的生物量、磷含量分别下降10%、15%，间作玉米分别下降29%、29%。2)当根系间距较近时，蚕豆的根际pH低于玉米，且间作玉米和蚕豆之间的根际pH存在显著差异，而在根系间距较远的处理中两作物的根际pH无差异；相邻作物根系间距较远时，间作玉米的根际有机酸浓度显著高于单作玉米，间作玉米与单作玉米的根际有机酸浓度在根系间距较近时无差异。3)不同处理中单作和间作玉米的总根长、总根表面积无差异。综上所述，在低磷条件下，单作玉米的种内竞争强度随根系间距的增大而降低；而在不同根系间距下，间作玉米与蚕豆的种间竞争强度相似且无种间磷吸收促进作用。因此，种间根际互作形成的磷吸收竞争作用或促进作用依赖于适度的土壤磷供应。     

Multiple chromosome triplication in Liatris

A plant with four and one with six extra A chromosomes were discovered in a hybrid swarm involving Liatris aspera and Liatris spicata. In each plant, the extra chromosomes formed trivalents with chromosomes of the normal complement. This pairing behavior indicates that of the ten chromosomes in the complement the plants are triplicate for four and six chromosomes, respectively.     

普通小麦与华山新麦草衍生后代的农艺性状和细胞遗传学研究

对中国春ph2b突变体(CSph2b)与华山新麦草属间杂种F1的衍生后代(BC2和BC1F1)进行了细胞学观察和田间农艺性状调查。其主要结果为:①染色体数目从2n=42-52之间变化,其中2n=45出现频率最高,为20.27%,而2n=51最低,仅有0.66%;②大部分植株有较多单价体;③在染色体数目较多的植株中,能够观察到三价体、四价体、落后染色体、三分孢子体、微核以及染色体桥;④部分植株表现高抗条锈病和赤霉病;⑤各个性状的变异系数都大于亲本的变异系数,分蘖的变异系数最大;⑥2n=42和2n=44的植株细胞学相对稳定,为获得华山新麦草染色体的附加系、代换系或易位系等小麦遗传育种材料奠定了基础。     

玉米矮缩病危害的损失测定与防治指标研究

为明确玉米矮缩病发生程度与玉米产量损失之间的关系,2011—2013年在浙江省临安市和桐庐县系统调查了玉米矮缩病在主栽品种‘浚单20’、‘浚单18’上的发生、危害及产量损失情况。结果表明：与健株相比,感病玉米植株均表现株高下降、节间缩短、叶长变短、叶宽变窄、果穗缩短、实粒减少、粒重下降;发病程度为1~4级的病株生物量下降28.76%~81.31%,单穗籽粒产量分别下降32.29%~98.91%。玉米植株发病率为2.31%~39.44%,产量损失率为1.03%~32.13%,两者呈显著相关性。通过回归分析建立了危害损失关系式,根据现有玉米生产条件和效益水平,计算出经济允许损失水平并制订了相应的防治指标。     

Ovipositional responses of Spodoptera frugiperda on host plants provide a basis for using Bt-transgenic maize as trap crop in China

Spodoptera frugiperda, the pest fall armyworm(FAW), is widespread in more than 100 countries. To date, planting insectresistant transgenic crops is one of the main control methods in its native countries. In this study we evaluated Bt-transgenic maize(Bt maize) and non-transgenic(conventional) maize and six other host plants in greenhouse pot experiments and field trials for oviposition preference by the Chinese populations of FAW. In laboratory trials, female moths preferred to oviposit on maize with no significant preference between conventional and Bt maize. However, after conventional and transgenic maize were exposed to FAW larvae and damaged, oviposition was significantly higher on transgenic maize than on the conventional one. Field trials showed that for plants less damaged at an early stage(seedling stage), oviposition of FAW on transgenic and conventional maize was significantly higher than that on wheat, sorghum, foxtail millet, peanut and soybean while showing no significant difference between transgenic or conventional maize. FAW adults mainly laid eggs on Bt maize, while the larval density and leaf damage rating or percentage of damaged plants were significantly lower than on conventional maize. Larval density and its damage on conventional maize were significantly higher than that on Bt maize and the other five hosts. Thus, maize is a highly preferred and suitable host for S. frugiperda feeding and ovipositing, and Bt maize can be used as trap crop to protect other crops.     

铵硝态氮单独供应对苗期淹水胁迫玉米氮磷钾累积的影响

【目的】研究不同供氮水平铵态氮（NH₄⁺-N）、硝态氮（NO₃^--N）对苗期淹水胁迫玉米氮、磷、钾含量及累积的影响,为采取氮营养调控途径优化玉米的氮、磷、钾营养特性及提高玉米的抗涝性提供理论依据。【方法】采用室内砂砾培养及模拟淹水胁迫的方法,研究2个玉米品种（“利民15”和“皖玉9号”）分别在苗期单独供应不同水平（3,7 mmol/L）NH₄⁺-N和NO₃^--N 时,根、茎、叶中氮、磷、钾含量及累积量对淹水胁迫的响应。【结果】在2种氮形态和供氮水平下,淹水胁迫分别对低供NH₄⁺-N处理（A3F）和高供NO₃^--N处理（N7F）苗期玉米生长的影响相对较小,且淹水胁迫对根系生长的抑制效应明显大于地上部。在NH₄⁺-N营养条件下,提高氮素供应水平对苗期淹水胁迫玉米氮、磷、钾的累积无显著影响,但根系氮含量显著增加而钾含量显著降低;在NO₃^--N营养条件下,提高氮素供应水平则可显著提高苗期淹水胁迫玉米对氮、磷、钾的累积与吸收。【结论】在模拟淹水胁迫条件下,提高NH₄⁺-N供应水平可明显增加苗期玉米根系含氮量并降低其根系含钾量,导致氮、钾比例失调,进而降低其耐淹性;但提高NO₃^--N供应水平则可明显增加苗期玉米对氮、磷和钾的累积与吸收,相对增强了玉米的耐淹性。     

玉米果穗出子率QTL定位及上位性分析

利用郑58×昌7-2组配的225个F2单株为作图群体,构建了全长为1 987.7 cM、覆盖玉米基因组10条染色体的分子标记遗传连锁图,标记间平均距离11.0 cM.采用复合区间作图方法(CIM),对玉米出子率性状进行QTL定位,利用多区间作图法(MIM)对上位性效应进行了分析.结果表明,在玉米第1染色体和第3染色体上检测到2个稳定的QTL位点,分别可以解释8.47%和10.52%的表型遗传变异.检测到5对上位性QTL,涉及6个位点,分布于第1,2,3,4和第5染色体,共解释9.94%表型遗传变异.这说明除了加性效应和显性效应外,上位性效应也是出子率性状的重要遗传基础.     

玉米骨干自交系品质基因SSR标记遗传多样性研究

选用玉米品质基因第2条染色体上的2对和第7条染色体上的3对SSR引物,对32个玉米自交系材料进行遗传多样性分析。结果表明:不同引物在不同自交系间多样性范围与程度都不相同,umc1066具有最好的多态性,带型稳定,重复性好。在o2基因内的SSR标记区域变异具有独立性,并非保守序列。具有o2基因的自交系与不具有o2基因的自交系,在基因的等位区段具有明显差异;在都具有o2基因的自交系间o2基因区段内的SSR标记具有多样性;具有o2基因的材料与优良自交系之间具有较好的多态性。