优质高产大花生新品种潍花6号的选育研究

潍花6号是潍坊市农科院以79266做母本，鲁花11号为父本进行有性杂交，系统选育而成。经1998—2001年山东、河北两省区试、生产试验及高产示范表明，该品种具有早熟、高产稳产、果大仁大、出仁率高、耐贮性好、口味佳等突出特点，2001年和2002年分别通过山东、河北两省农作物品种审定委员会审定，是一个适于中国北方花生产区栽培的优质高产大花生新品种。     

Developing gene-tagged molecular markers for functional analysis of starch-synthesizing genes in rice (Oryza sativa L.)

The granule-bound starch synthase(GBSS), starch branching enzymes 1 (SBE1)and 3 (SBE3) are major enzymes involved in starch biosynthesis in rice endosperm. Available sequences of Sbe1 and Sbe3 genes encoding corresponding SBE1 and SBE3 have been used to identify homologous regions from genome databases of both the indica rice 93-11 and the japonica rice Nipponbare. Sequence diversities were exploited to develop gene-tagged markers to distinguish an indica allele from a japonica allele for both Sbe1 and Sbe3 loci. With these newly developed gene-tagged markers and available Wx gene markers, the roles of these starch-synthesizing genes (Sbe1, Sbe3, and Wx) in determining amylose content (AC) in the rice endosperm were evaluated using a double-haploid (DH)population derived from a cross between the indica rice cv. Nanjing11 and the japonica rice cv. Balilla. Only the Wx and Sbe3 loci had significant effects on the AC variation. On average, indica Wx ^a genotypes showed ∼9.1% higher AC than japonica Wx ^b genotypes, while indica Sbe3 ^a genotypes showed ∼1.0% lower AC than japonica Sbe3 ^b genotypes. A significant interaction was also observed between Wx and Sbe3 loci whereby the amylose content was 0.3% higher in Sbe3 ^a than Sbe3 ^b genotypes in the presence of the Wx ^a allele, but it was lower by 2.3% in the presence of the Wx ^b allele. Overall, polymorphisms at the Wx and Sbe3 loci together could account for 79.1% of the observed AC variation in the DH population. The use of gene-tagged markers in marker-assisted selection and gene functional analysis was also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Factors influencing tissue culture responses of mature seeds and immature embryos in turf-type tall fescue

In an effort to optimize tissue culture responses of turf‐type tall fescue for genetic transformation, the effects of five culture medium supplements on tissue culture responses were investigated with immature embryos and mature seeds of an elite cultivar, ‘Coronado’, as explant tissues. For both explants, calli induced on 6‐benzylaminopurine (BAP)‐containing medium had significantly improved regeneration ability. The optimal concentration of BAP for the induction of regenerable callus from mature seeds was 0.1 mg/l. Thidiazuron also improved callus regeneration frequency of mature seeds. Casein hydrolysate, L‐proline and myo‐inositol improved callus induction of immature embryos but not mature seeds. They did not improve callus regeneration frequencies with either explant tissue. By simply slicing the mature seeds into two halves longitudinally, the callus induction frequencies, as well as the corresponding overall plant regeneration frequencies, were increased approximately three‐ to six‐fold in all three combinations of 2,4‐dichlorophenoxyacetic acid and BAP in callus induction media.     

大白菜细胞质雄性不育的分子鉴定及序列分析

为了获得3种大白菜细胞质雄性不育系Ogu CMS,Pol CMS,CMS96和保持系间的多态性以及定位大白菜CMS96不育系所属的不育类型,利用设计的atp6,orf222单一和混合引物PCR扩增3组11份同核异质大白菜细胞质雄性不育系和保持系mtDNA。结果表明,atp6引物在大白菜Ogu CMS不育系扩增的200 bp片段为其特异带;orf222引物仅在大白菜Pol CMS和CMS96不育系有扩增产物,但二者有3点完全不同:大白菜Pol CMS不育系扩增产物为675bp,CMS96不育系扩增产物为669 bp,二者相差6个核苷酸,后者定名为大白菜CMS96-orf222。大白菜CMS96-orf222与甘蓝型油菜Nap CMS的nad5c基因和Nap-orf222基因同源性均为99%,E值为0.0;大白菜Pol CMS的675 bp序列具有ORF224开放阅读框,没有保守结构域,而大白菜CMS96的669 bp序列具有ORF222开放阅读框和保守结构域YMF19。另外,atp6和orf222混合引物多重PCR扩增产物存在明显多态性:800 bp为大白菜保持系的差异带型;2 300 bp和1 500 bp为大白菜Pol CMS不育系特异带型;200 bp为大白菜Ogu CMS不育系特异带型;690 bp为大白菜CMS96不育系特异带型。该方法仅用一次PCR反应快速地将3种大白菜细胞质雄性不育系和保持系一次性全部区分开,为大白菜分子育种和常规育种更好地相结合提供了简单、快速、准确和重复性好的方法和手段。     

Cell lines and plants obtained after protoplast fusions of Rosa+Rosa,Rosa+Prunus and Rosa+Rubus

Summary Protoplasts of three Rosa cultivars were fused with each other, with protoplasts of Prunus `Colt' and with protoplasts of Rubus laciniatus, using polyethylene glycol 4000 as a fusogen. Protoplasts of Prunus were incapable of cell division and those of Rosa and Rubus were disabled by treatments with metabolic inhibitors, either iodoacetate (IOA) or rhodamine 6-G (R6G). Parental protoplasts were then fused in combinations that required complementation for their survival. RAPD analysis of 41 fusion-derived cell lines showed that two lines resulting from fusions of Rosa + Rosa and one from a fusion of Rosa + Prunus, contained some DNA markers from both fusion partners. The others contained markers of only one fusion parent. This showed that after protoplast fusion, the heterokaryons did not develop into cell lines with stable hybrid nuclei. Plants regenerated from cell lines derived from Rosa + Prunus and Rosa + Rubus fusions contained DNA markers of only Rosa and their DNA amounts were no greater than that of the Rosa parent. However, they differed morphologically from the Rosa parent to a remarkable degree, possibly because they inherited undetected genes of Prunus or Rubus, or because they were somaclonal variants of the Rosa parent. Alternative strategies for the production of somatic hybrids are discussed.     

茶树叶绿素合成相关基因克隆及在白叶1号不同白化阶段的表达

高等植物叶绿素的生物合成对其正常光合作用起关键作用。本文根据前期芯片杂交结果, 采用RT-PCR和RACE技术克隆了3个茶树叶绿素合成相关基因, 分别为谷氨酸-tRNA还原酶(CsGluTR)、叶绿素合酶(CsChlS)、叶绿素酸醋氧化酶(CsCAO), 对应的GenBank的登录号为HQ660371、HQ660370、HQ660369。序列分析表明, CsGluTR基因全长2165 bp, 开放阅读框长1665 bp, 编码554个氨基酸, 推测的蛋白分子量约为60.6 kD, 理论等电点为8.78;CsChlS基因全长1463 bp, 其中开放阅读框长1125 bp, 编码374个氨基酸, 推测的蛋白分子量约为40.5 kD, 理论等电点为8.58;CsCAO基因全长2146 bp, 其中开放阅读框长1611 bp, 编码536个氨基酸, 推测的蛋白分子量约为60.8 kD, 理论等电点为8.03。比对分析表明, 3个基因编码的氨基酸序列与其他植物中同源基因的相似性均在70%以上。利用荧光定量PCR技术检测3个基因在不同白化阶段的表达,表明, CsChlS和CsCAO基因具有明显的表达协同性, 它们在叶片中的表达量与叶片的颜色变化高度同步;而CsGluTR在白化叶片和正常叶片中的表达差异相对较小, 同时在新生芽叶转绿过程中最先恢复正常表达水平。说明在白化叶片中, 叶绿素的合成机制受到较大影响, 叶绿素合成受阻导致的叶片内色素类物质含量降低或消失是叶片白化的直接原因。     

旱种水稻结实期茎中碳同化物的运转及其生理机制

以杂交水稻汕优63和籼稻扬稻6号为材料，研究了旱种(地膜覆盖栽培)水稻结实期茎中碳同化物的运转及其生理机制。结果表明，抽穗至成熟期旱种水稻标记14C从茎中向籽粒的再分配、茎中非结构性碳水化合物(NSC)的运转率及其对籽粒的贡献率均显著高于水种(常规栽培)水稻。水稻旱种后籽粒中的蔗糖合成酶和酸性转化酶活性、茎中α-淀     

Characterization of waxy grain sorghum lines in relation to granule-bound starch synthase

The waxy phenotype, associated with endosperm containing little or no amylose, has been recognized in sorghum (Sorghum bicolor L. Moench) since 1933. Although variants of the waxy gene are well characterized in other cereals, the waxy trait has been assumed to be controlled by a single allele, wx, in sorghum. Recent improvements in technologies encourage re-examination of the waxy sorghums. The objectives of this research were therefore to identify and characterize sorghum lines with differing waxy alleles and to describe the actions of those alleles in crosses. Grain of eight waxy sorghum lines (BTxARG1, BTx630, Tx2907, B.9307, 94C274, 94C278, 94C289, 94C369), three wild-type checks (BWheatland, RTx430, BN122), and F₂ families from crosses among a subset of these lines were evaluated for presence or absence of granule-bound starch synthase (GBSS), the gene product of the wx locus, and wild-type vs. waxy endosperm. The F₂ segregation ratios were tested for fit to a 3:1 ratio using Chi-square analyses. Two distinctly different naturally occurring waxy alleles were identified: One with no GBSS (GBSS−), and one with apparently inactive GBSS present (GBSS+). We propose that the waxy allele with no GBSS be designated wx^a, and that waxy allele with apparently inactive GBSS present be designated wx^b. These two alleles are located in close proximity on the waxy locus. The wx^b allele is dominant to the wx^a allele in terms of GBSS production, and both are recessive to the wild-type Wx in terms of amylose content. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

水稻灌浆期籽粒中3个与淀粉合成有关的酶活性变化

在水培和盆栽条件下，研究了6个水稻品种(含籼/粳杂交组合、新株型品系)灌浆期强、弱势粒中ADPG焦磷酸酶(EC2.7.7.21)、淀粉合成酶(EC2.4.1.21)和淀粉分枝酶或Q-酶(EC2.4.1.18)的活性变化及其与灌浆充实的关系。3个酶的活性变化与籽粒灌浆动态相关联：淀粉酶最高活性出现的时间稍前或同步于最大灌浆速率的时间；ADPG焦磷酸酶