Generation of the genetic mutant population for the screening and characterization of the mutants in response to drought in maize

The autonomous Mutator(Mu)transposon in maize(Zea mays L.)lines 115F,330I,and 715D were crossed with inbred lines B73,Mo17,97108,and H9-21,M1were self-pollinated to establish the Mu insertion-mutagenized M2seeds pool and The M2plants encompassed a large amount of biological variation relevant to improving agronomy traits by observed in the field.Next we statistically analyzed these candidate mutants.Under drought stress,we screened seedlings at 3-leaf stage by using the infrared thermography,and successfully got 108 droughtinsensitive and 121 drought-sensitive candidate mutants from more than 38,000 lines,which temperature were significant different with others.These candidate mutants were primarily analyzed by infrared thermography,chlorophyll fluorescence,leaf water losing,photosynthetic characteristics,content of soluble sugars,soluble protein,and proline assay.The selected mutants were cloned by MuTAIL–PCR methods.Here we provide the better genetic materials for research on maize breeding for drought tolerance.     

五种基因型玉米籽粒灌浆特性的研究

通过对五种基因型玉米在授粉后３０天内籽粒灌浆过程的研究，认为不同类型的玉米籽粒灌浆差异主要表现在量上。普通玉米，糯性玉米的灌浆速度快，强度大，失水也快；甜玉米相对地灌浆速度慢，失水也慢。授粉后１０－２０天是各类型玉米籽粒鲜重，干重增加的最大时期。     

玉米单细胞培养的研究

将类型Ⅱ愈伤组织从固体培养基转入悬浮培养,发现在改良1/2MN培养基,1.0-2.0mg/L的2,4-D浓度及1000mg/L肌醇存在下,适合所有不同基因型玉米自交系悬浮细胞的生长和细胞成活率的提高.悬浮培养5-6天继代1次是适宜的.大量元素减半和0.5-1.0mg/L的2,4-D浓度用于玉米单细胞培养,能获得良好的细胞分裂和生长.细胞悬浮培养获得了愈伤组织.     

玉米细胞悬浮系建立和单细胞培养影响因素的研究

以玉米幼胚为外植体进行离体培养,获取愈伤组织,建立悬浮细胞系.试验结果表明:基因型无论是对细胞悬浮系建立还是对单细胞培养都有很大影响;改良1/2B5培养基用于细胞悬浮培养能获得良好的细胞生长与分裂;继代培养时间以及大量元素减半都影响玉米悬浮细胞的生长.     

玉米矮花叶病毒外壳蛋白基因的克隆研究

玉米(Zea mays L.)矮花叶病在国内外广泛发生,且在玉米生产中造成了重大损失.通过RT-PCR法从具有典型的玉米矮花叶病症状的玉米叶片中克隆了外壳蛋白(Coat protein,CP)基因,测序和同源性比较表明所克隆的CP基因来自玉米矮花叶病毒(Maize dwarf mosaic virus,MDMV)B株系,全长920个碱基对,开放阅读框编码219个氨基酸,该基因可进一步用于玉米抗矮花叶病的转基因研究,以获得生产应用的抗病材料.     

外源甜菜碱对两种玉米耐盐性影响的研究

以玉米沈单和农大108为实验材料,研究了外源甜菜碱对盐分胁迫条件下玉米(Zea may)幼苗水势、叶绿素含量、丙二醛(MDA)含量、细胞膜透性、cNa /cK 等的影响.结果表明,外源甜菜碱能够提高玉米幼苗叶片水势、叶绿素含量,降低膜透性、丙二醛含量,参与多种代谢调节,影响作物的耐盐性.另外,外源甜菜碱可以增强根系对Na ,K 的选择吸收能力,促进质膜主动排Na 过程,降低盐胁迫下叶片中Na 含量.     

玉米自交系幼胚离体培养及植株再生研究

采用4种不同类型的玉米自交系为供试材料，用MS和AG培养基进行诱导培养。获得胚性愈伤组织，结果表明，基因型和外植体的发育时期对愈伤组织的诱导率有很大影响；不同的2，4-D浓度及山梨醇浓度影响愈伤组织的保持和质量改善；6-BA浓度为2mg／L时，有利于再分化培养，在再分化培养中，获得了再生植株。     

一个新的水稻矮秆突变体sd-sl的遗传与基因定位研究

新的矮秆基因的发掘、研究和利用对水稻育种和植物生长发育机制研究有重要的作用.用60Coγ射线辐照粳稻9522,获得一个能稳定遗传的突变体.该突变体表型为株高较野生型矮,叶片短而微卷.将该突变体与籼稻广陆矮杂交,F2代呈3∶1分离,说明该突变体受隐性单基因控制.通过InDel分子标记对F2代分离群体进行遗传定位,将该基因定位于第6染色体InDel标记OS604附近.随后又发展了多对有多态性的InDel分子标记,将该基因座位精细定位在InDel标记XL6-6和XL6-1之间,AP003490和AP005619上,两个引物之间的物理距离为118 kb.本研究为该克隆基因及其作用机理的探究奠定了基础.     

玉米健壮素对玉米支持根影响的初步研究

经玉米健壮素处理的玉米支持根发育与脱氢酶活性增强,氨基酸合成增多。这些变化对提高玉米抗倒伏和吸收水肥能力有利,在玉米生产实践上也拟可用于延长生育后期根系活动期和防止早衰。     

微卫星标记分析籼粳亚种间的遗传多样性

208对引物中具有多态性的引物123对,占所用引物的59．13％,不同染色体的微卫星分析的多态性不同,染色体9,10微卫星的多态性高于其它染色体,染色体12上的微卫星标记的多态性最差,仅为46．15％．聚类分析表明,所有的供试材料可分为两群,即籼稻群和粳稻群,聚类结果与亲本材料亲缘关系基本一致,说明微卫星标记能较好地区分籼稻和粳稻,由于农艺性状是基因表达的结果,易受环境影响,聚类结果不能从整体上充分反应品种间的遗传变异,42份常用杂交水稻亲本材料聚类分析表明,恢复系和不育系遗传基础均较狭窄,但恢复系和不育系之间的遗传距离相对较远,从一定程度上反映了遗传距离与杂种优势正相关。     

一个水稻抗纹枯病突变体的遗传分析及其基因的初步定位

高水平抗纹枯病突变体和高感纹枯病品种蜀恢881杂交构建分离群体,经F2分离世代的遗传分析,抗、感单株比例符合3 1(χc2=0.563,χ12,0.05=3.84),初步确定该突变体对纹枯病的抗性由一对显性主效基因所控制,命名为Rsb-2(t)。利用已合成的530对微卫星引物,对抗纹枯病突变体和蜀恢881进行多态性引物筛选,用多态性引物对上述F2分离群体的全部感病单株和部分抗病单株的DNA进行PCR分析,借助MAPERMAKER/EXP3.0软件,对其微卫星标记实验数据进行连锁分析,将Rsb-2(t)定位于第3染色体的p臂,发现RM218、RM251、RM4321和RM5748与Rsb-2(t)连锁,它们均位于着丝粒端,连锁距离分别为32.1 cM,41.1 cM,42.4 cM和49.7 cM。研究结果为进一步对该基因的精细定位奠定了基础。     

玉米黄色素的提取和分析

使用超临界CO2法从水解玉米蛋白粉中提取玉米黄色素.通过单因素实验和正交实验优化了超临界CO2萃取玉米黄色素的工艺条件,并通过HPLC检测提取物中玉米黄素和叶黄素的含量.结果表明:超临界CO2提取玉米黄色素的最佳条件为:萃取时间2 h,CO2流速35 kg/h,萃取压力30 MPa,萃取温度45℃,夹带剂15%无水乙醇,此时玉米黄色素产量为210.97 μg/g.     

Construction of cDNA library from iron-deficiency induced maize roots and screening and identification of iron-stress gene Fdr3

To isolate Fe-deficient related (Fdr) genes, an expression cDNA library of 4.5×105 pfu/μg has been constructed from maize roots in iron-stress. 6 clones have been screened from the cDNA library by differential hybridization screening. It is proved that an Fdr3 cDNA clone expressed stronger under iron-deficient condition than under iron-sufficient one by Northern blot and Western blot.     

不同性别类型黄瓜品种基因组DNA的提取及RAPD标记的初步研究

采用改良的异丙醇沉淀法成功地提取了11个不同性别类型黄瓜品种的基因组DNA.以此作为PCR扩增反应模板进行RAPD分析,结果从22个10bp随机寡核苷酸引物中筛选出2个引物S23和S91在全雌性品种MT700和其他性别类型的品种之间稳定地扩增出多态性.至于该多态性标记是否与黄瓜雌性性别有关,还有待进一步分析.     

Relationship between differ- ential gene expression pat- terns in functional leaves of maize (Zea mays L.) at milk filling stage and heterosis using cDNA-AFLP

The genetic mechanism of heterosis has been one of the most challenging subjects in life science since the 20th century. However, because of the complexity of its genetic basis and the limitations of research methods and means, there was no substantial progress in the research of heterosis from the beginning of the 20th century to the 1990s. The development in molecular biology and bio- technology, especially that in quantitative trait loci (QTL) mapping methods, has made it possible to resea…     

墨962群体主要农艺性状的遗传特征分析

对热带玉米群体墨962主要农艺性状进行了综合分析,结果表明:墨962群体遗传基础较遗传变异丰富,抗病、抗逆性强,丰产性突出.在进行群体籽粒产量改良时,百粒重的选择意义不大,而穗粗不受其他性状选择的干扰.在墨962群体的利用方面,应注重低穗位、长穗、高出籽率的选择.     

小麦温光生态研究及其在生产中的应用进展

文章对小麦温光反应理论,温度和光照在小麦生长和发育过程中的相互关系,以及温光反应理论在生产实践中的应用新进展进行了总结,可为小麦温光生态的研究和小麦生产实践提供理论指导。     

Detection of quantitative trait loci and heterotic loci for plant height using an immortalized F2 population in maize

A set of recombinant inbred lines (RIL) derived from Yuyu22, an elite hybrid widespread in China, was used to construct an immortalized F2 (IF2) population comprising 441 different crosses. Genetic linkage maps were constructed containing 10 linkages groups with 263 simple sequence repeat (SSR) molecular markers. Twelve and ten quantitative trait loci (QTL) were detected for plant height in the IF2 and RIL populations respectively, using the composite interval mapping method, and six same QTL were identified in the two populations. In addition, ten unique heterotic loci (HL) located on seven different chromosomes were revealed for plant height using the mid-parent heterosis as the input data. These HL explained 1.26%―8.41% of the genotypic variance in plant height heterosis and most expressed overdominant effects. Only three QTL and HL were located in the same chromosomal region, it implied that plant height and its heterosis might be controlled by two types of genetic mechanisms.     

一种新型天蚕素杂合肽AD和aFGF突变体融合蛋白S的构建及表达

目的：构建和表达一种带有凝血酶切割位点的天蚕素杂合肽AD和人酸性成纤维细胞生长因子(acidic fibroblast growth factor,aFGF)突变体的融合蛋白S,在创伤皮肤表面应用时能自动裂解成有抗感染功能的天蚕素杂合肽AD和皮肤修复功能的aFGF突变体。方法：利用P(R技术分别扩增出带有凝血酶切割位点的天蚕素杂合肽AD基因和aFGF突变体基因。再利用两作为模板,用PCR方法扩增出编码融合蛋白S的DNA。将该DNA片段插入到载体pPICZaA中,并利用电转法将质粒转入毕赤酵母smd1168中,Zeocin抗性筛选重组转化子。甲醇诱导72h,SDS-PAGE电泳检测融合蛋白S的表达,并用Western-blot杂交检测融合蛋白S的免疫原性。结果：筛选出的重组转化株在甲醇诱导后能表达相对分子质量约为19000的融合蛋白S,而该蛋白能与aFGF抗体产生免疫反应。结论：用PCR方法获得编码含凝血酶切割位点的天蚕素杂合肽AD和aFGF突变体的融合蛋白SDNA,并在毕赤酵母中实现了表达。     

Genetic analysis and fine mapping of an enclosed panicle mutant esp2 in rice （Oryza sativa L.）

The phenomenon of panicle enclosure in rice is mainly caused by the shortening of uppermost internode.Elucidating the molecular mechanism of panicle enclosure will be helpful for solving the problem of panicle enclosure in male sterile lines and creating new germplasms in rice.We acquired a monogenic recessive enclosed panicle mutant,named as esp2 (enclosed shorter panicle 2),from the tissue culture progeny of indica rice cultivar Minghui-86.In the mutant,panicles were entirely enclosed by flag leaf sheaths and the uppermost internode was almost completely degenerated,but the other internodes did not have obvious changes in length.Genetic analysis indicated that the mutant phenotype was controlled by a recessive gene,which could be steadily inherited and was not affected by genetic background.Apparently,ESP2 is a key gene for the development of uppermost internode in rice.Using an F 2 population of a cross between esp2 and a japonica rice cultivar Xiushui-13 as well as SSR and InDel markers,we fine mapped ESP2 to a 14-kb region on the end of the short arm of chromosome 1.According to the rice genome sequence annotation,only one intact gene exists in this region,namely,a putative phosphatidylserine synthase gene.Sequencing analysis on the mutant and the wild type indicated that this gene was inserted by a 5287-bp retrotransposon sequence.Hence,we took this gene as a candidate of ESP2.The results of this study will facilitate the cloning and functional analysis of ESP2 gene.