Genetic mapping and utilization of quantitative trichome-mediated insect resistance in potato

Introgression of trichome-mediated insect resistance from the wild speciesSolanum berthaultii has become a major focus of the potato improvement program at Cornell University during the past twelve years. Several quantitative characters are involved in this resistance which is effective against a wide range of pest types. Correlative biochemical assays have been developed to assay specific components of the resistance, and the effects of the resistance on the target pests have been studied. Quantitative laboratory assays and specific measurements of insect behavior and biology have increased the precision of selection and enable the investigation of the genetic control of the resistance.We are currently using restriction fragment length polymorphisms (RFLPs) for genetic mapping of factors controlling the trichome traits fromS. berthaultii. Backcrosses to both the wild and the cultivated species parents have been evaluated for phenotypes contributing to the resistance mechanism, including trichome density, sucrose ester and polyphenol oxidase production by the trichomes, and the enzymatic browning reaction responsible for insect entrapment. Genetic maps are being developed for these progenies, using RFLP markers previously mapped in potato. Field and greenhouse trials under insect infestations are also being conducted with the mapping progeny. Our goal is to locate genes responsible for quantitative insect resistance by correlating RFLP variation at mapped loci with the trichome phenotypes and insect resistance. Genetic markers for these traits will be useful in transfer of the effective wild chromosomal segments into and among tetraploid potatoes, and for a better understanding of the resistance mechanism.     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

陕甘宁老区脆弱生态环境定量评价——以榆林、延安两市为例

位于黄土高原中部的陕甘宁老区生态环境极为脆弱 ,近年来由于气候、人类开发资源等自然和人为原因 ,使生态环境的脆弱程度升高。选定年降水量、年均温、蒸发量等 8个指标 ,定量评价各县 1 970 - 2 0 0 0年的脆弱度状况 ,结果表明榆林、延安两市生态环境整体脆弱 ,脆弱度存在空间差异但差异不明显 ,时间段上的波动幅度不大。陕甘宁老区脆弱生态环境具有不稳定性 ,对外界干扰较敏感。     

Further Contributions to the Development of a Differential Set of Pea Cultivars (Pisum sativum) to Investigate the Virulence of Isolates of Aphanomyces euteiches

Common root rot (Aphanomyces euteiches Drechs.) has become a very destructive disease in the French pea crops since 1993. For an accurate investigation of the virulence variability among French A. euteiches populations and between French and foreign populations, a new set of differential pea genotypes was developed. Thirty-three American and European pea lines, displaying different levels of resistance, were screened in a growth chamber against two French isolates. Symptoms (disease severity from 0 to 5, evaluating symptom surface on roots and epicotyl) and percentage of top fresh weight (inoculated/uninoculated top fresh weight ratio) were measured. From this screening 12 relatively resistant lines, from various genetic backgrounds, were identified along with a highly susceptible control. This set of 13 genotypes was inoculated under controlled conditions with 14 isolates from France, Sweden, USA, Canada and New Zealand, to investigate genotype–isolate interactions. Root symptoms were rated (disease severity), and a susceptibility/resistance threshold was established at disease severity = 1. Significant quantitative interactions were observed, and five 'resistance patterns' were identified, leading to a set of six pea genotypes: Baccara (susceptible), Capella, MN313, 902131, 552 and PI180693. Fields trials of this set in 1999 and 2000 gave the same resistance rankings than in growth chamber conditions. This set will allow more accurate assessments of the variability in virulence/aggressiveness of A. euteiches isolates from France and foreign countries, and further investigations of the epidemiological and genetic basis of pea–A. euteiches interactions.     

谷物胚乳性状QTL区间作图的贝叶斯方法

将贝叶斯统计原理和胚乳性状的数量遗传模型相结合,以分离群体中各植株的分子标记基因型以及植株上若干粒种子胚乳性状的单粒观测值为数据模式,提出胚乳性状QTL区间作图的贝叶斯方法.该方法通过Gibbs以及Metropolis-Hastings抽样实现的马尔科夫链蒙特卡罗(MCMC)算法获得QTL效应和位置的估计.方法的有效性用染色体水平和基因组水平2套模拟方案进行验证,结果表明:贝叶斯方法能够准确地估计胚乳性状QTL的位置和效应,并同时区分2种显性效应.     

Taqman探针实时定量PCR检测猪伪狂犬病毒

选取猪伪狂犬病毒(PRV)基因组中gE基因,设计特异性引物和Taqman探针,利用实时定量PCR来定量检测PRV.利用PCR技术扩增猪伪狂犬病毒gE基因80 bp片段,并克隆到pMD18-T载体上,阳性重组质粒命名为pgE80.pgE80质粒进行10倍系列稀释,作为荧光PCR检测的标准模板,定量拷贝数,并绘制标准曲线.以提取的PRV、猪圆环病毒2型(PCV2)、猪细小病毒(PPV)、猪瘟病毒(CSFV)、猪繁殖与呼吸障碍综合症病毒(PRRSV)和传染性胃肠炎病毒(TGEV)的DNA作为模板,进行特异性检测.该实时定量PCR方法,标准曲线斜率为-0.37,R2=0.992,可以检测到20个拷贝数.PRV能被扩增出S型荧光曲线,而其它病毒均未能扩出.用PRV强毒肌肉注射仔猪,定量PCR比普通PCR更能灵敏而特异地检出呼吸道排毒和血液带毒.建立的实时定量PCR方法可用于临床样品快捷、准确、方便地检测.     

利用DH群体进行白菜株高和开展度的QTL定位分析

应用AFLP、SRAP、RAPD、SSR及同工酶等标记构建的326个标记位点的白菜遗传图谱和81个DH株系群体,采用M apQTL 5软件对白菜的株高和开展度进行QTL定位。结果表明：通过2年试验,在10个连锁群上共检测到27个QTL,主要分布在R 5、R 10连锁群上：其中控制株高的有11个,控制开展度的有16个;获得2年稳定表达的控制株高的QTL 1个,控制开展度的QTL 3个;各QTL加性效应各不相等,其遗传贡献率为13.3%-29.2%;控制株高和控制开展度的QTL位点表现为紧密连锁或相似的位置,主要集中在R 5连锁群上。     

白背飞虱酚氧化酶原PPO基因特性及其免疫应答

【目的】酚氧化酶(phenoloxidase,PO)是昆虫体内的免疫蛋白,在昆虫体内起重要的调节作用,主要以无活性的酚氧化酶原(prophenoloxidase,PPO)形式存在。本研究在转录组测序获得白背飞虱(Sogatella furcifera)3条PPO序列的基础上,通过分析白背飞虱体内不同SfPPO的发育和组织表达模式,并进一步抑制SfPPO的表达以及病原菌诱导,探讨SfPPO在白背飞虱体内的生物学功能。【方法】以白背飞虱3条SfPPO序列为研究对象,利用生物信息学方法分析其蛋白结构以及与其他昆虫之间的同源性。并以注射绿色荧光蛋白(green fluorescent protein,GFP)的dsRNA作为对照组,通过注射合成的ds SfPPO后采用实时荧光定量PCR(qRT-PCR)检测基因表达抑制效果;此外,为了探究SfPPO在白背飞虱生长发育和免疫应答方面的作用,利用qRT-PCR检测SfPPO在不同发育阶段及组织中的表达情况,以及注射大肠杆菌(Escherichia coli)和枯草芽孢杆菌(Bacillus subtilis)后3个SfPPO的诱导表达情况。【结果】...     

基于实例分割的白羽肉鸡体质量估测方法

针对白羽肉鸡体质量测量自动化水平低、易造成肉鸡应激的问题,提出一种结合深度学习的非接触式白羽肉鸡体质量估测方法.利用Mask R-CNN和YOLACT(You only look at coefficients)两种实例分割算法获取白羽肉鸡位置与覆盖掩膜,并进行效果对比;采用自适应掩膜随机提取白羽肉鸡身体部分边缘点,并...