The threat of wild habitat to scab resistant apple cultivars

Evaluations of plant resistance to pathogens are rarely made using isolates from wild habitats, although the heterogeneity of such habitats may generate pathogen diversity which could be a source of new virulence in cultivated habitats. The aim of this study was to investigate whether scab resistance factors, identified and characterized in apples using isolates of Venturia inaequalis from a cultivated habitat, remained effective against isolates from a wild habitat. Three V. inaequalis core collections originating from the cultivated apple Malus × domestica and from two wild species, M. sieversii and M. sylvestris, were established to maximize pathogen diversity. For each core collection, 10 isolates were inoculated in mixtures onto 51 genotypes from an apple progeny segregating for two qualitative resistance genes and six quantitative resistance loci (QRL). On each apple genotype, isolates that contributed to the scab symptoms were identified within the mixture using microsatellite markers. The most frequently detected isolates were inoculated singly to compare their aggressiveness according to their host origin. The results showed that isolates from a wild habitat were able to infect the susceptible apple genotypes. However, these isolates were never more aggressive than isolates from the cultivated habitat on the resistance factors tested. It can therefore be concluded that the resistance factors used in this study, identified with V. inaequalis isolates from a cultivated habitat, remained effective against isolates from M. sylvestris and M. sieversii.     

大豆不同生育时期叶绿素含量QTL的定位及其与产量的关联分析

利用来自波高×南农94-156的151个RI家系检测与4个不同生育时期叶绿素含量（累积量、净增量）有关的QTL,并分析其与籽粒产量、表观生物学产量和表观收获指数的关系。结果表明,与叶绿素累积量有关的QTL位于D1a+Q、F、G、H、L和M连锁群上,每个QTL可解释表型变异的6.9%~23.4%。V6和R2期没有检测到2个年份均表达的QTL,而在R4期检测到4个在2个年份均表达的QTL（qccF.1、qccG.2、qccH.1和qccM.1）,R6期仅检测到1个QTL（qccH.1）在2个年份均表达,该QTL在R4也表达。与叶绿素含量净增量有关的QTL位于B2和L连锁群上,在V6-R2时期没有检测到与叶绿素净增量有关的QTL,在B2和L连锁群上的两个QTL（qccB2-1.1和qccL.1）在R2-R4和R4-R6时期均表达,qccB2-1.1可解释表型变异的6.4%~9.8%,而qccL.1所解释表型变异达29.5%~31.3%。但这两个QTL在R2-R4和R4-R6时期表达的性质不同,且与2年均表达的籽粒产量QTL共位。这印证了生育后期叶绿素含量与籽粒产量间存在的极显著正相关。     

Quantitative trait loci for phytate in rice grain and their relationship with grain micronutrient content

Phytate (inositol-hexa-phosphate) has an important role in plants but it also may have anti-nutritional properties in animals and humans. While there is debate within the plant breeding and nutrition communities regarding an optimum level in grain, there appears to be little information at the molecular level for the genetics of this trait, and its association with important trace elements, in particular, Fe and Zn. In this preliminary study, quantitative trait loci (QTL) for grain phytates, Zn and Fe in glasshouse-grown rice lines from an IR64 × Azucena doubled haploid population were identified. Correlations between phytate and essential nutrients were also studied. Transgressive segregation was found for most traits. Phytate and total P concentrations had one QTL in common located on chromosome five with the (high concentration) allele contributed from Azucena. There were significant positive correlations between phytate and inorganic phosphorus (P), total P, Fe, Zn, Cu and Mn concentrations for both grain concentration and content. However, the QTLs of phytate were not located on the same chromosomal regions as those found for Fe, Zn and Mn, suggesting that they were genetically different and thus using molecular markers in breeding and selection would modify the phytate level without affecting grain micronutrient density.     

A genetic linkage map in southern‐by‐spring oat identifies multiple quantitative trait loci for adaptation and rust resistance

We developed 178 recombinant inbred lines from a southern‐by‐spring oat population designated as “TxH.” These lines were genotyped to generate a high‐quality linkage map that resolved 6,902 markers into 21 linkage groups that matched closely with the latest hexaploid oat consensus map. Three major quantitative trait loci (QTLs) affecting heading date were found in locations that are consistent with known QTLs and candidate genes, and two other QTLs affecting heading date were found in novel locations. Five QTLs affecting plant height were found. Both sets of QTLs are responsible for transgressive segregation observed for these two traits. Four QTLs affecting resistance to crown rust, caused by the pathogen Puccinia coronata f. sp. avenae, were identified. Two of these QTLs are consistent with known clusters of rust resistance genes, while two may represent new locations of novel rust resistance genes. A complete set of SNP sequences suitable for generating markers for molecular selection is provided.     

山东栒子全长转录组测序数据组装及功能注释

山东栒子是中国山东省的特有种,现已处于极度濒危状态。目前,山东栒子的分子生物学研究较少,基因数据库资源极度缺乏,急需探究其生物学遗传信息,以加快对山东栒子的保护遗传学工作。本研究以山东栒子叶片、花、成熟果实为实验材料,利用PacBio Sequel测序平台对其转录组进行全长转录组测序。共得到高质量去冗余的转录本53932个,作为最终转录本序列。预测到的CDS区共52490个;对其SSR位点进行分析,对测序得到Unigenes进行单核苷酸至六核苷酸重复的SSR位点搜索,共搜索到26796个SSR位点。对非冗余转录本利用BLAST软件与NR、Swissprot、GO、COG、KOG、KEGG 6个数据库进行比对,一共成功注释了53319个Unigenes,其中与NR数据库进行比对中注释为苹果的的相关基因数量最多,其次是白梨和桃;在GO数据库中有33305条山东栒子Unigenes被注释分类,由生物学过程、细胞成分和分子功能三部分组成;在与COG数据库比对中,共有23910条比对到了同源序列,且一共被分为25类;在与KEGG数据库的一系列比对中,可将Unigenes映射到126条代谢通路中。本研究在高通量全长转录组水平对山东栒子进行了系统研究,这为进一步开展山东栒子的分子标记开发和挖掘优良基因提供了科学依据,从而推动山东栒子的保护与利用。     

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。     

小麦粒长和粒宽的QTL定位分析

【目的】粒长、粒宽是小麦种子重要的形态性状,该性状对籽粒的外观商品品质、产量及磨粉品质均至关重要,研究不同环境条件下小麦粒长、粒宽的单个标记和复合区间作图的QTL定位,对小麦粒长、粒宽的分子标记辅助选择具有重要参考作用。【方法】应用一个由115个系组成的W7984/Opata 85重组自交系(RIL)群体,建立了由394个DNA分子标记组成的遗传连锁图,在2种不同环境条件下对小麦粒长、粒宽进行了单个标记的回归分析和复合区间作图的QTL定位。【结果】在单个标记的回归分析中检测到5个粒长的QTLs、3个粒宽的QTLs;复合区间作图分析结果表明,控制粒长的QTLs分别位于5BL和7DS上,在5BL上的贡献率为20.20%~20.81%,LOD值为4.50~4.55;在7DS上的贡献率为13.54%~13.91%,LOD值为2.94~3.20。控制粒宽的QTL位于2B上,贡献率为13.71%~19.30%,LOD值为2.98~4.18。【结论】位于5B和7D上的控制粒长的QTL和位于2B上的控制粒宽的QTL在2种条件下均能检测到。     

玉米和水稻全基因组控制重组频率QTL的遗传定位分析

 【目的】尝试利用分子标记和QTL定位技术直接定位影响重组频率的QTL,探索物种遗传变异难易的分子基础。【方法】分别借助3张玉米和3张水稻分子标记连锁图,以所有染色体上标记的交换次数为性状进行分析。【结果】分别定位了7个和11个影响玉米和水稻重组频率的QTL。以玉米和水稻高密度分子标记连锁图IBM302和Genetic98每条染色体上标记的交换次数为性状,分别在玉米和水稻上定位了12个和57个QTL。【结论】影响重组频率的基因真实存在。培育高重组频率材料将有助于加速基因的定位和克隆、比较图位克隆基因和遗传育种进程。     

Quantitative trait loci for fruit size and flowering time-related traits under domestication and diversifying selection in cucumber (Cucumis sativus)

In cucumber, the genetic basis of traits under domestication and/or diversifying selection is not well understood. Here, we reported QTL mapping for flowering time and fruit size-related traits with segregating populations derived from a cultivated × wild cross. Phenotypic data of flowering time (FT), fruit size (FS), fruit number (FN) and fruit weight per plant (FW) were collected in multiple environments. QTL analysis identified 19 QTL for these traits. We found that the major-effect QTL FT1.1 played an important role in regulating flowering time in cultivated cucumber, whereas the minor-effect QTL FT6.3 contributed to photoperiod sensitive flowering time during domestication. Two novel consensus FS QTL, FS1.4 and FS2.3, seem to be the targets of selection during breeding for the US processing cucumber. All other FS QTL were co-localized with previously detected QTL using populations derived from cultivated cucumbers, suggesting that they were under selection during both initial domestication and subsequent improvement. Results from this study also suggested that the wild cucumber is a useful resource for capturing positive transgressive segregation and novel alleles that could be explored in cucumber breeding.     

多位点基因打靶技术的鳜鱼体内实验研究

在转基因动物的研究中,一般采用随机插入的方法。基因在受体动物的基因组中随机整合,严重制约了转基因动物外源基因的稳定遗传,是转基因动物研究亟待解决的问题。转基因定点整合技术,即基因打靶技术是解决上述问题的重要途径,但是基因打靶存在打靶效率太低的问题。本研究小组已建立了体外动物细胞的多位点基因打靶技术,并构建了用于鳜鱼的体内多位点基因打靶的打靶载体,本文开展多位点基因打靶技术的鳜鱼体内实验研究。以鳜鱼rDNA基因(编码rRNA的基因)及其间隔序列作为同源重组引导序列,构建含干扰素基因的打靶载体,以其间隔序列为靶位点,针对鱼类胚胎发育特点,采用精子介导技术进行多位点基因打靶,最终获得定点整合干扰素基因的鳜鱼,并建立一套适用于动物的基因定点敲入的体内基因打靶技术。主要研究内容如下:(1)采用组织培养方法,获得鳜鱼头肾淋巴细胞和脾淋巴细胞,采用脂质体介导法使多位点基因打靶载体在鳜鱼的细胞上实行基因转移。采用正负筛选方法,利用筛选药物G418和GCV(更昔洛韦)筛选转染后的细胞两周后,对存活细胞进行DNA水平和RNA水平鉴定。结果证明了干扰素基因在鳜鱼细胞上实现了定点整合和稳定表达。(2)利用精子介导法将经线性化处理后的多位点基因打靶载体在鳜鱼的胚胎上实行基因转移。(3)根据靶位点定点整合的正负筛选原理,利用筛选药物G418和GCV筛选与富集定点整合后的阳性鱼胚或鱼苗,结果成功孵化出106尾转基因鳜鱼鱼苗。(4)培育6个月后,取20尾转基因鳜鱼进行鉴定。采用PCR、RT-PCR、测序等方法进行定点整合的鉴定。结果表明:6尾检测到已转入的基因——干扰素基因,其中有3尾实现了定点整合并表达。本研究率先将多位点基因打靶技术应用到转基因鱼的研究中,建立一套适用于鱼类的高效、安全的多位点基因打靶技术,并获得外源基因定点整合并表达的鳜鱼。