Molecular detection of insecticide resistant alleles

The summary discusses techniques used to investigate mechanisms of resistance in the Colorado potato beetle (Leptinotarsa decemlineata) to several classes of pesticides.     

水稻纹枯病抗性关联分析及抗性等位变异发掘

采用苗期微室接种鉴定法,用144个分布于水稻全基因组的多态性标记,利用TASSEL软件GLM (Q)、MLM (Q+K)和MLM (PCA+K) 3种模型对456份水稻材料组成的自然群体进行纹枯病抗性关联分析。结果发现,有13个标记位点至少在两种模型中均被检测到与纹枯病抗性显著关联,单个位点可解释表型变异的1.84%~8.42%;其中10个标记位点位于以往报道的连锁定位的抗纹枯病QTL附近,3个标记位点(RM1036、RM5371和RM7585)是未曾报道的新的抗病相关位点。抗性等位变异RM7585-150对纹枯病发病病级减效效应最大;有259份材料携带抗性等位变异RM5371-129,占供试材料总数的56.8%,只有26份材料携带抗性等位变异RM1036-82,占供试材料总数的5.7%。水稻纹枯病发病病级与其含有的抗性等位变异数量呈极显著的负相关。本研究结果将为水稻抗纹枯病分子标记辅助育种提供理论依据。     

微卫星标记BM3413和OB_2在5个绵羊群体中多态性的研究

选萨福克羊、德国美利奴羊、特克塞尔羊、乌珠穆沁羊以及右玉本地绵羊5个绵羊群体共250只,通过耳组织提取基因组DNA,用2对微卫星引物进行PCR扩增,通过电泳分型、凝胶成像系统分析各位点等位基因及全部个体的标记基因型,计算基因频率、多态信息含量(PIC)和杂合度等,从分子水平上分析5个绵羊品种的遗传多态性.结果表明:BM3413和OB2这2个位点的等位基因数分别为7和8,多态信息含量PIC＞0.5,杂合度为0.827 1～0.846 7,均属于高度多态性位点,用于作为与生产性能相关的遗传标记是较理想的.     

核不育复等位基因向娃娃菜中转育的研究

根据"复等位基因遗传假说",以甲型两用系AB12为不育源(直筒生态型),采用杂交、自交、兄妹交的方法,将核不育基因向娃娃菜可育品系06006中转育,获得了不育株率100%的新核基因雄性不育系,扩大了原有不育系的遗传基础,拓宽了该优良雄性不育基因的应用范围.     

藏獒血液蛋白多态性研究

［目的］了解藏獒血液蛋白基因座的遗传变异状况,为藏獒品种资源保护及合理开发利用提供理论依据。［方法］采用不连续垂直板聚丙烯酰胺凝胶电泳技术,对河曲藏獒、青海藏獒、青海藏狮犬和青海土种犬共4个群体103只犬的7个血液蛋白基因座（Tf、Po、Sα2、Hb、Alb、Pr、Amy）的多态性进行研究,并分析不同群体的群体内遗传变异。［结果］ 4个犬群中,Tf、Po和Sα2 3个基因座上存在多态性,其中Tf和Po分别由3个等位基因控制,Sα2 由2个等位基因控制,Hb、Alb、Pr和Amy基因座均呈现单态;藏獒群体的有效等位基因数（Ne）和Nei氏平均预期基因杂合度（H）分别为1.532 4和0.230 3,均高于其他犬群。［结论］ 藏獒群体内在血液蛋白位点上存在丰富的遗传变异。     

Determination of the Number of SSR Alleles Necessary for the Analysis of Genetic Relationships Between Maize Inbred Lines

The amount of molecular marker information has considerable impact on the results of studies of crop germplasm genetic relationships in crop. The number of alleles required to reveal genetic relationship in maize inbred lines is a theoretical issue that needs to be addressed. In this study, 112 pairs of SSR （simple sequence repeat） primers and 97 maize inbred lines were selected to study the relationship between the number of inbred lines and the number of SSR primers and alleles required for a stable cluster. The results showed that the number of SSR primers is not tightly associated with the stability of the cluster analysis results, while an increase in the number of alleles can significantly improve the stability of cluster analysis results. The number of inbred lines （X） is significantly associated with the number of alleles required for stable cluster analysis （Y）, and the regression equation is Y- 600.8xe（-15.9/x）. This equation can be used to calculate the number of SSR alleles required for a genetic relationship study of maize inbred lines. These results provide a reference for determining of SSR alleles number in genetic relationship analysis of maize inbred line and other crop germplasm.     

The tryptophan-rich domain of puroindoline is directly associated with the starch granule surface as judged by tryptic shaving and mass spectrometry

The starch granule surface is a frontline of microbial attack and defence, operating in the background of normal starch granule metabolism. Puroindoline, a wheat protein which binds starch granule surfaces, contains a unique tryptophan-rich domain likely responsible for this property, though direct evidence is lacking. To test puroindoline’s tight association, prime starch granule extracts were water-washed 8 or 20 times and residual puroindoline removed using a solution of 50% isopropanol/50 mM NaCl. We found that this solvent was consistent in the amount of protein extracted from wheat flour and washed starch, regardless of initial protein content. Relative quantification of puroindoline following water-washing was performed using dot blot. Washing more than 8 times did not further reduce puroindoline content of starch granules suggesting a strong association with the starch granule surface. To identify the tryptophan-rich domain tightly associated with the starch granule surface, a combination of in situ tryptic digestion and mass spectrometry was used. Following digestion and water-washing, 50% isopropanol/50 mM NaCl was used to remove tightly-associated peptides for identification by mass spectrometry. Using this method, we identified the tryptophan-rich domain of puroindoline directly bound to the starch granule surface of wheat.     

Puroindoline genotype,starch granule size distribution and milling quality of wheat

Genetically-diverse wheat samples from the Australian Winter Cereals Collection propagated in two environments were sequenced to identify puroindoline genotypes then the relationships between flour yield, genotype, starch granule size distribution and starch-bound puroindoline protein content were investigated. The Pina-D1a, Pinb-D1b genotype resulted in a higher average flour yield than either the Pina-D1b, Pinb-D1a or the Pina-D1a, Pinb-D1a but the ranges of flour yields for the three genotypes showed considerable overlap. For both hard wheat genotypes (Pina-D1a, Pinb-D1b or Pina-D1b, Pinb-D1a), a higher proportion of type A to type C starch granules was associated with higher flour yield and this relationship accounted for between 31% and 33% of the variation in flour yield. This result is consistent with previously reported findings for soft wheat. For the Pina-D1a, Pinb-D1b genotype, increased flour yield was also associated with a decrease in starch granule-bound puroindoline protein, which accounted for 31–35% of the variation in flour yield across the two environments. The combined effect of starch granule type and associated puroindoline content accounted for 68% of the variation in flour yield within the Pina-D1a, Pinb-D1b genotype.     

Functionality of native tetraploid wheat starches: Effects of waxy loci alleles and amylose concentration in blends

Partial waxy (reduced amylose) and fully waxy (amylose-free) tetraploid durum wheats (Triticum turgidum L. var. durum) were used to investigate the relationships between both intra- and inter-granular variation in amylose concentration and starch functionality. Starches isolated from each genotype (intra-granular amylose variants) were compared to those of commercially available hexaploid wild-type and waxy starches, and functionalities compared to blends (inter-granular variants) of durum waxy and wild-type starches of 0, 6, 12, 18, 24, & 30% amylose content. Starch particle size distributions were not significantly different amongst partial waxy and wild-type genotypes; waxy samples had significantly smaller mean starch granule size. Few significant differences for crystallite melting and related intrinsic heat as determined by differential scanning calorimetry (DSC) were observed. The detected differences in starch gel color or gel strength generally were observed for the waxy samples relative to wild-type. Numerous significant differences were observed via Rapid-Visco Analysis (RVA). Pasting peak viscosity and breakdown were inversely proportional to % amylose. Wx-B1 null final viscosity differed significantly from that of all other blends and genotypes, demonstrating that genotypic differences exist amongst the partial waxy types, independent of amylose concentration per se.