小麦与高冰草体细胞杂种F2代Ⅱ.同工酶和蛋白质分析

比较了小麦与高冰草杂种F2代不同穗行和株系的部分植株与亲本进行酯酶同工酶及可溶性蛋白质,分析高冰草遗传物质在杂种后代的遗传和表达,结果证明高冰草遗传物质在杂种中稳定存在并正常表达.粗蛋白含量的测定表明,杂种F1及F2代蛋白质含量为17.49%～20.18%,介于亲本小麦(15.14%)与高冰草(25.58%)之间.氨基酸分析发现,杂种F2代种子的几种必需氨基酸,包括缬氨酸、蛋氨酸、异亮氨酸、赖氨酸、组氨酸和精氨酸,显著高于普通小麦.     

151份小麦品种（系）中主要品质基因的分子检测

明确江苏瑞华农业科技有限公司151份小麦品种（系）中品质基因的分布情况,利用分子标记技术对小麦谷蛋白亚基（HMW-GS）、多酚氧化酶（PPO）活性和1B/1R异位系进行检测。结果表明：高分子量谷蛋白亚基（HMW-GS）基因频率Ax2为1.32%,Dx5为17.22%,By8为70.20%,Bx7为89.40%,Ax2+Dx5、Ax2+By8和Ax2+Bx7均为0.66%,Dx5+By8为7.28%,Dx5+Bx7为14.57%,By8+Bx7为62.25%;多酚氧化酶（PPO）活性相关基因频率Ppo-A1b为45.70%,Ppo-D1a为25.83%,双低PPO活性等位基因Ppo-A1b+Ppo-D1a为优异等位基因,频率为8.60%;1B/1R异位系的分布频率为86.75%;同时携带低PPO活性等位变异组合,且为非1B/1R异位系的材料只有2份,频率为1.32%;同时携带谷蛋白亚基Bx7和低PPO活性等位变异组合,且为非1B/1R异位系的材料只有1份,可作为小麦育种材料。     

小麦与高冰草体细胞杂种F_2代Ⅱ.同工酶和蛋白质分析

比较了小麦与高冰草杂种Ｆ２代不同穗行和株系的部分植株与亲本进行酯酶同工酶及可溶性蛋白质,分析高冰草遗传物质在杂种后代的遗传和表达,结果证明高冰草遗传物质在杂种中稳定存在并正常表达．粗蛋白含量的测定表明,杂种Ｆ１及Ｆ２代蛋白质含量为１７．４９％～２０．１８％,介于亲本小麦（１５．１４％）与高冰草（２５．５８％）之间．氨基酸分析发现,杂种Ｆ２代种子的几种必需氨基酸,包括缬氨酸、蛋氨酸、异亮氨酸、赖氨酸、组氨酸和精氨酸,显著高于普通小麦．     

小麦与高冰草体细胞杂种F2代：Ⅱ.同工酶和蛋白质分析

比较了小麦与高冰草杂种Ｆ２代不同穗行和株系的部分植株与亲本进行酯酶同工酶及可溶性蛋白质,分析高冰草遗传物质在杂种后代的遗传和表达,结果证明高冰草遗传物质在杂种中稳定存在并正常表达。粘粘白含量的测定表明,杂种Ｈ１及Ｆ２代蛋白质含量为１７．４９％～２０．１８％,介于亲本小麦（１５．１４％）与高冰草（２５．５８％）之间。氨基酸分析发现,杂种Ｆ２代种子的几种必需基酸,包括缬氨酸、蛋氨酸、异亮氨酸、赖氨酸、     

普通小麦与高冰草不对称体细胞杂种F2代：Ⅰ表型与性状 …

普通小麦济南１７７原生质体和经紫外线照射的高冰草原生质体用ＰＥＧ法诱导融合,形成杂种植株,但未能正常结实。杂种植株经子房诱导产生可育Ｆ０代;将Ｆ０代产生的４粒种子播种后产生Ｆ１代植株,对由其产生的Ｆ２代４个株系进行田间观察和室内考种。分析表明,Ｆ２代植株的表现型及产量性状均明显优于亲本小麦。田间穗行,株系表型稳定,具有抗寒、茎杆硬、抗倒伏等优良性状。在分蘖数、单株粒重衣千粒重等产量性状上亦有明显优     

普通小麦与高冰草不对称体细胞杂种F_2代 Ⅰ表型与性状分析

普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）济南１７７原生质体和经紫外线照射的高冰草（Ａｇｒｏｐｙｒｏｎｅｌｏｎｇａｔｕｍ）原生质体用ＰＥＧ法诱导融合，形成杂种植株，但未能正常结实．杂种植株经子房诱导产生可育Ｆ０代；将Ｆ０代产生的４粒种子播种后产生Ｆ１代植株，对由其产生的Ｆ２代４个株系进行田间观察和室内考种．分析表明，Ｆ２代植株的表现型及产量性状均明显优于亲本小麦．田间穗行、株系表型稳定，具有抗寒、茎杆硬、抗倒伏等优良性状．在分蘖数、单株粒重及千粒重等产量性状上亦有明显优势，此研究为培育体细胞杂种优良小麦品系奠定了基础．     

小麦与高冰草不对称体细胞杂种F5代部分株系的花粉母细胞染色体观察分析

普通小麦（Triticum aestivum L.)济南177原生质体（受体）和经紫外线照射的高冰草（Agropyron elongatum)原生质体（供体）用PEG法诱导融合，形成外形偏向小麦的不对称体细胞杂种及后代，经过田间繁育，现已到F5代，在对F1至F4代杂种植株完成各项分析的基础上，对对F5代不同株系（Ⅱ-2，8-1，Ⅱ-Ⅰ-8）的花粉母细胞染色体进行观察和分析，结果表明，杂种花粉母细胞在减数分裂时基本正常，88％以上的细胞染色体数目在20－21对之间，并且绝大多数染色体可以正常配对形成二价体，少数细胞存在小染色体，数目多为1－2个，且大部分以配对形式存在，由此可见，杂种在遗传上基本是稳定的，且杂种的表型及遗传在较短的世代就基本稳定，但个别杂种花粉母细胞中也存在减数分裂异常现象，如不均等分离，落后染色体，染色体桥、多着丝粒体，易位环及多价体等。     

超大穗小麦高分子量麦谷蛋白亚基组成分析

采用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳法(SDS-PAGE)分析了超大穗小麦高分子量麦谷蛋白亚基(HMW-GS)组成及其F1代的遗传多样性。结果表明,9份超大穗小麦中共检测到7种类型的高分子量麦谷蛋白亚基,在Glu-A1位点,出现了两种亚基,null和1亚基,Glu-D1位点只有2 12一种亚基,而Glu-B1位点上亚基类型最丰富,有4种类型,尤其出现了两种新型的复合亚基组合形式7 9 8和7 9 14 15。遗传分析表明,在超大穗小麦与普通小麦杂交后代F1中,双亲的高分子量麦谷蛋白亚基呈显性遗传,且显母性遗传倾向。     

两个杂交组合中转基因小麦外源1Dx5基因的遗传

利用杂交组合川89-107×B72-8-11b和绵阳26×B72-8-11b的亲本、F1、F2代,研究转基因小麦B72-8-11b中外源品质基因1Dx5表达的遗传.结果表明:外源1Dx5基因在F1代中呈现显性,在F2代中呈现3(有):1(无)的分离,有功能拷贝整合在1个位点,遵从孟德尔遗传模式,这对杂交育种选择策略的制订具有指导意义.     

转1Dx5基因小麦突变株系的研究

以转1Dx5基因小麦3个高分子量麦谷蛋白突变株系B73-6-1-1、B73-6-1-2和B73-6-1-3及转基因受体L88-6为材料,对其主要农艺性状、低分子量麦谷蛋白表达情况及染色体核型进行研究.结果表明:在田间栽培条件下,纯合稳定转基因突变株系之间及其与转基因受体之间主要农艺性状存在差异,其中株高和千粒重存在显著差异(P=0.05);同时,SDS-PAGE结果显示,在小麦低分子量麦谷蛋白区域,突变株系之间无明显差异,而突变株系与转基因受体之间部分区域表达量存在明显差异并产生一个突变条带;通过核型分析比较,得出突变株系之间及其与转基因受体之间,染色体无论是相对长度还是臂比值都没有显著差异.     

Transgene inheritance and quality improvement by expressing novel HMW glutenin subunit （HMW-GS）genes in winter wheat

The expression vector pBPC30, which carries the high molecular weight glutenin subunit (HMW-GS) 1Dx5 and 1Dy10 genes, was transferred into hexaploid winter wheat cv. Jinghua No. 1, Jing411 and Jingdong No. 6 explants of immature embryos and immature inflorescence by particle bombardment. A large number of resistant transgenic plants were obtained under the selection of herbicide bialaphos or phosphinothricin (PPT). Confirmed transgenic plants of To generation showed successful integration of HMW-GS genes and bar gene into the wheat genome. T1 generation of transgenic plants can resist 20--150 mg/L PPT.Protein analysis of T2 seed by SDS-PAGE showed that HMW-GS 1Dx5 and 1DylO genes were well expressed in offspring seed of transgenic lines by co-expression with or substitution of endogenous 1Dx2 or 1DylO. In one transgenic line, TG3-74, a new protein band between endogenous protein subunits 7 and 8 (marked as 8*) of glutenin appeared,but endogenous subunit 8 (encoded by 1By8 gene) was absent. Analysis of gluten rheological quality on seed proteins of 102 T3 plants showed that the sedimentation value of 5 transgenic lines (44.2149.0 mL) was remarkably improved,59.6%---64.3% higher than that of wild type Jinghua No. 1 and Jingdong No. 6, similar to bread wheat Cheyenne (48.0 mL). Analysis of dough rheological properties of transgenic lines showed that the dough stable time of 5 transgenic lines range from 16 to 30 min, whereas the dough stable time of wild type was only between 3--7 min. Our research suggests that introducing novel HMW-GS genes into wheat is an efficient way to improve its bread-making quality.     

小麦与高冰草原生质体融合及再生能力的恢复

以普通小麦（Ｔｒｉｔｉｃｕｍａｅｓｔｉｖｕｍ）济南１７７悬浮细胞来源的原生质体与高冰草（Ａｇｒｏｐｙｒｏｎｅｌｏｎｇａｔｕｍ）愈伤组织来源的原生质体用ＰＥＧ法诱导融合．由于来源材料的长期继代，小麦原生质体的再生能力已很低；高冰草原生质体不能分裂；而融合产物却能高频率的分化出完整植株．融合再生植株的表型类似高冰草，染色体数目和同工酶谱亦然．但它们早期发育的模式与小麦相似．     

Molecular identification of asymmetric somatic hybrids between wheat and Italian ryegrass

Sexual incompatibility between common wheat and Italian ryegrass was an obstacle for transferring useful traits from italian ryegrass to wheat. In order to use those desirable genetic resources to improve wheat and to create new cytoplasmic germplasm, the protoplasts of wheat and Italian ryegrass were successfully electrofused and the somatic hybrid plants were regenerated. Examination with 6 restriction enzymes, 13 probes including 9 mtDNA probes (H454, Pst24, B30, Pro I, 490, B342, pHJ2-7-1, B376, 7), 3 cpDNA probes (pHvc p1, pHvc p5 and pHvc p8) and one nuclear DNA probe-- pTA71 (rDNA) in total 73 enzyme/ probe combinations revealed rich polymorphism between the fusion partners. RFLP analysis indicated that approximately 93.4% of the regenerated plants were true somatic hybrids. AFLP analysis implied that the somatic hybrids were highly asymmetric. The RFLP analysis using mt- and cpDNA specific probes also demonstrated the non-coexistence of mitochondria and chloroplasts from the fusion partners in the somatic hybrid cells.     

转基因小麦T1代中外源基因的检测和分析

该研究采用Multiplex PCR和SDS-PAGE技术对转1 Dx5基因(ORF)T1代小麦进行了检测和分析.结果显示,Multiplex PCR能够扩增出转基因T1代材料中1 Dx2基因和1 Dx5基因的特征片段,表明外源基因已整合到受体基因组中;SDS-PAGE检测到一新的蛋白质亚基X,表明外源基因的插入引起了转基因T1代籽粒中HMW-GS组成的变化.该研究不仅验证了线性基因片段遗传转化策略的可行性,而且印证了普通小麦Glu-1D等位基因的多重PCR分子标记体系的有效性,为培育安全型的转基因作物新种质打下坚实的基础,加快小麦分子育种进程.     

普通小麦与玉米的体细胞杂交再生完整植株

从小麦济南177制备得到两种原生质体，一种来源于具有一定分化能力的愈伤组织但其分裂能力较低；一种来源于生长迅速的悬浮细胞但不能分化，将二种原生质体混合，共同作为受体与经过紫外线（UV）照射的玉米（Zea mays L)原生质体在PEG诱导下融合，培养后获得再生克隆并进一步分化为植株，而它们单独与玉米原生质体融合均不能再生植株，通过染色体、同工酶及5SrDNA分析及生长习性分析证明再生植株均为体细胞杂种。     

春小麦谷蛋白亚基、醇溶蛋白电泳谱带及主要品质指标的主成分分析和聚类分析

对18份小麦品种(系)的高分子量谷蛋白亚基、醇溶蛋白电泳谱带的组成及主要常规品质指标进行了检测，并对31个权重最大的小麦品质影响因子进行了主成分分析和聚类分析。结果表明，对小麦品质贡献大的前10个因子的先后次序为：湿面筋含量、硬度、Glil7．6、Gli22、沉淀值、Gli30、1Dx5 1Dy10、蛋白质含量、面筋指数、Gli59．0。这10个因子既相互促进，又相互制约。在小麦品质育种中要高度重视谷蛋白亚基及醇溶蛋白电泳谱带的作用。18个小麦品种可以聚成5类，10个主成分值在5类品种中的表现差异很大。因此，在小麦品质育种的亲本选配时，既要注意协调10个主要品质影响因子的关系，又要使同一组合亲本间保持一定的遗传距离。     

Genetic basis of heterosis and inbreeding depression in rice （Oryza sativa L.）

The genetic basis of heterosis was studied through mid-parent, standard variety and better parent for 11 quantitative traits in 17 parental lines and their 10 selected hybrids in rice (Oryza sativa L.). The characters were plant height, days to flag leaf initiation, days to first panicle initiation, days to 100% flowering, panicle length, flag leaf length, days to maturity, number of fertile spikelet/panicle, number of effective tillers/hill, grain yield/10-hill, and 1000-grain weight. In general the hybrids performed significantly better than the respective parents. Significant heterosis was observed for most of the studied characters. Among the 10 hybrids, four hybrids viz., 17Ax45R, 25Ax37R, 27Ax39R, 31Ax47R, and 35Ax47R showed highest heterosis in 10-hill grain yield/10-hill. Inbreeding depression of F2 progeny was also studied for 11 characters of 10 hybrids. Both positive and negative inbreeding depression were found in many crosses for the studied characters, but none was found significant. Selection of good parents was found to be the most important for developing high yielding hybrid rice varieties.     

The relationship of differential expression of genes in GA biosynthesis and response pathways with heterosis of plant height in a wheat diallel cross

Heterosis in internode elongation and plant height is commonly observed in hybrid plants, but the molecular basis for the increased internode elongation in hybrids is unknown. In this study, midparent heterosis in plant height was determined in a wheat diallel cross involving 16 hybrids and 8 parents, and real-time PCR was used to analyze alterations in gene expression between hybrids and parents. Significant heterosis of plant height and the first internode in length were observed for all 16 hybrids, but the magnitude of heterosis was variable for different cross combinations. Analysis revealed that the heterosis of the first internode was significantly correlated to that of plant height （r = 0.56, P 〈 0.05）, suggesting that the increased elongation of the first internode is the major contributor to the heterosis in plant height. Real-time PCR analysis exhibited that significant difference in heterosis of gene ex- pression was observed among all cross combinations. Moreover, heterosis of the first internode in length was correlated significantly and positively with expression heterosis of KS, GA3ox2-1, GA20ox2, GA20ox1D, GA-MYB and GID1-1, but significantly and negatively with expression heterosis of GAI and GA2ox-1, which is consistent with our recently proposed model of GAs and heterosis in wheat plant height, suggesting the alteration of GA biosynthesis and response pathways might be responsible for the observed heterosis in plant height.     

应用基因组原位杂交鉴定杂交后代中的簇毛麦染色体

用地高辛（Digoxigenin-11-dTup)标记的簇毛麦染色体组DNA为探针，以普通小麦“中国春”总DNA作封阻进行基因组荧光原位杂交，对小麦和簇毛麦杂交和回交后代进行检测。结果显示，在杂交后代的28条染色体中有7条簇毛麦染色体，在发生部分染色体加倍的回交代后鉴定出含7条簇毛麦染色体重的易位系。GISH的准确鉴定以及易位系的获得为向小麦导入簇志麦的有用基因提供了宝贵材料。     

高粱总DNA导入春小麦新品系高分子量麦谷蛋白亚基的变化

通过花粉管通道法将高粱总DNA导入春麦甘麦8号、陇春13号和陇春10号,经过多代选择获得了5个稳定遗传新品系.在高分子量麦谷蛋白亚基分析中,甘麦8号后代89144的高分子量麦谷蛋白亚基发生突变,较其受体多了5 10亚基,而少了2 12亚基;其他几个转基因后代与其受体比较,高分子量麦谷蛋白亚基组成未发生变化;但是各亚基的相对质量分数有较大变化.高分子量麦谷蛋白亚基的组成和各亚基质量分数的变化直接影响小麦品质.本研究对外源总DNA花粉管通道法导入小麦在改良小麦品质方面的作用进行了讨论.