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。     

Quantitative trait loci associated with lodging tolerance in soybean cultivar ‘Toyoharuka’

Lodging tolerance (LT) is an important trait for high yield and combine-harvesting efficiency in soybean [Glycine max (L.) Merr.]. Many previous studies have investigated quantitative trait loci (QTLs) for lodging score (LS) in soybean. Most of the investigated QTLs were located in the proximal region of maturity or growth habit loci. The aim of this study was to identify genetic factors for LT not associated with maturity or growth habit. QTL analysis was performed using a recombinant inbred line (RIL) population derived from a cross between ‘Toyoharuka’ (TH), a lodging-tolerant cultivar, and ‘Toyomusume’ (TM). The genotypes of TH and TM were estimated as both e1e2E3E4 and dt1. The average LS over 4 years was used for QTL analysis, identifying a major and stable QTL, qLS19-1, on chromosome 19. The LS of the near-isogenic line (NIL) with the TH allele at Sat_099, the nearest marker to qLS19-1, was significantly lower than the NIL with the TM allele at that position. The TH allele at Sat_099 rarely had a negative influence on seed yield or other agronomic traits in both NILs and the TM-backcrossed lines. Our results suggest that marker-assisted selection for qLS19-1 is effective for improving LT in breeding programs.     

Identification of molecular markers associated with sugar-related traits in a <Emphasis Type="Italic">Saccharum</Emphasis> interspecific cross

The cultivated sugarcane (Saccharum spp. hybrids, 2n = 100–130) is one crop for which interspecific hybridization involving wild germplasm has provided a major breakthrough in its improvement. Few clones were used in the initial hybridization event leading to a narrow genetic base for continued cultivar development. Molecular breeding would facilitate the identification and introgression of novel alleles/genes from the wild germplasm into cultivated sugarcane. We report the identification of molecular markers associated with sugar-related traits using an F₁ population derived from a cross between S. officinarum ‘Louisiana Striped’ × S. spontaneum ‘SES 147B’, the two major progenitor species of cultivated sugarcane. Genetic linkage maps of the S. officinarum and S. spontaneum parents were produced using the AFLP, SRAP and TRAP molecular marker techniques. The mapping population was evaluated for sugar-related traits namely, Brix (B) and pol (P) at the early (E) and late (L) plant growing season in the plant cane (04) and first ratoon (05) crops (04EB, 04LB, 04LP, 05EB and 05EP). For S. officinarum, combined across all the traits, a total of 30 putative QTLs was observed with LOD scores ranging from 2.51 to 7.48. The phenotypic variation (adj. R²) explained by all QTLs per trait ranged from 22.1% (04LP) to 48.4% (04EB). For S. spontaneum, a total of 11 putative QTLs was observed with LOD scores ranging from 2.62 to 4.70 and adj. R² ranging from 9.3% (04LP) to 43.0% (04LB). Nine digenic interactions (iQTL) were observed in S. officinarum whereas only three were observed in S. spontaneum. About half of the QTLs contributed by both progenitor species were associated with effects on the trait that was contrary to expectations based on the phenotype of the parent contributing the allele. Quantitative trait loci and their associated effects were consistent across crop-years and growing seasons with very few QTLs being unique to the early season. When the data were reanalyzed using the non-parametric discriminant analysis (DA) approach, significant marker-trait associations were detected for markers that were either identical to or in the vicinity of markers previously identified using the traditional QTL approach. Discriminant analysis also pointed to previously unidentified markers some of which remained unlinked on the map. These preliminary results suggest that DA could be used as a complementary approach to traditional QTL analysis in a crop like sugarcane for which saturated linkage maps are unavailable or difficult to obtain.     

Development and evaluation of pyramiding lines carrying early or late heading QTLs in the indica rice cultivar ‘IR64’

The heading date is an important trait for determining regional and climatic adaptability in rice. To expand the adaptability of the indica rice cultivar ‘IR64’, we pyramided multiple early or late heading quantitative trait locus (QTLs) in the ‘IR64’ genetic background by crossing previously developed near-isogenic lines (NILs) with a single QTL for early or late heading. The effects of pyramiding QTLs were observed in three different climatic zones of the Philippines, Madagascar, and Japan. The early heading pyramiding lines (PYLs) headed 6.2 to 12.8 days earlier than ‘IR64’ while the late heading PYLs headed 18.8 to 27.1 days later than ‘IR64’. The PYLs tended to produce low grain yield compared to ‘IR64’. The low yield was not improved by combining SPIKE, which is a QTL that increases the number of spikelets per panicle. Conversely, ‘IR64-PYL(7+10)’ carrying Hd5 and Hd1 headed earlier, produced more tillers, and more panicles per m² than ‘IR64’, and mitigated the yield decrease in early heading. These results suggest that the effects of pyramided QTLs on heading date were consistent across various environments and PYLs could be used to enhance the adaptation of ‘IR64’ in other rice growing environments.     

小麦白粉病成株抗性和抗倒伏性及穗下节长度的QTL定位

由小麦品种花培3号和豫麦57杂交获得了DH群体168个株系, 利用305个SSR标记对白粉病成株抗性、抗倒伏性和穗下节长度进行了QTL定位研究。DH群体及两亲本于2005年和2006年种植于山东泰安, 2006年种于安徽宿州。利用基于混合线性模型的QTLNetwork 2.0软件, 共检测到12个加性效应位点和10对上位效应位点。在4D染色体上控制白粉病成株抗性的qApr4D, 贡献率为20.0%, 在各环境中稳定表达, 其抗病等位基因来源于抗病亲本豫麦57; 在7D染色体上控制小麦穗下节长度的qIlbs7D, 贡献率为12.9%, 在各环境中稳定表达。加性效应和上位效应对小麦白粉病成株抗性、抗倒伏性和穗下节长度的遗传起重要作用, 并且基因与环境常常具有互作效应。以上两个QTL可分别用于小麦白粉病成株抗性和穗下节长度的分子标记辅助选择。     

Use of component analysis in QTL mapping of complex crop traits: a case study on yield in barley

Genes contributing to the quantitative variation of a complex crop trait can be numerous. However, using existing approaches, the number of quantitative trait loci (QTL) detected for a trait is limited. Therefore, rather than looking for QTL for a complex trait itself, determining QTL for underlying component traits might give more information. In this study the potential of component analysis in QTL mapping of complex traits was examined using grain yield in spring barley as an example. Grain yield was divided into three components: number of spikes/m², number of kernels/spike, and 1000‐kernel weight. These traits were measured for individuals of a recombinant inbred‐line population in field trials conducted over 2 years. By the use of an approximate multiple QTL model, one to eight QTL were detected for each trait in a year. Some QTL were mapped to similar positions in both years. Almost all QTL for yield were found at the position of or in close proximity to QTL for its component traits. A number of QTL for component traits were not detected when yield itself was subjected to QTL analysis. However, relative to the QTL for yield itself, all component‐trait QTL did not explain the variation in yield better. The results in relation to the potential of using component analysis in studying complex crop traits are discussed.     

Migration of gluten under shear flow as a novel mechanism for separating wheat flour into gluten and starch

This paper describes a novel principle for the separation of wheat flour into starch and gluten in a concentrated medium. The process is based on the use of simple shear flow in a cone-and-cone device. The separation takes place in two steps. Initially, local segregation of gluten and starch phases occurs, leading to formation of macroscopically visible gluten patches distributed throughout the dough. This local segregation can be understood by considering the dough as a visco-elastic matrix containing an inert filler (starch). Further shearing leads to aggregation of those patches and migration (large-scale separation) towards the apex of the cone. As a result, the wheat dough is separated into a protein-poor fraction, containing less than 4% protein, and a protein-rich fraction containing almost 50% protein on a dry weight basis. However, under the process conditions used, upon a very long shearing, a redistribution of the aggregated gluten structures in the starch phase was observed, demonstrating a processing limit for the separation performance. Compared to traditional processing, the separation process presented shows opportunities for producing high quality gluten accompanied with significant water savings. Considering the fact that simple shear flow in steady rate is less harmful to gluten quality, such a separation process could benefit gluten quality.     

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。     

粗山羊草D组染色体对小麦若干产量性状的影响

为研究粗山羊草对小麦产量性状的影响,对四倍体硬粒小麦与粗山羊草杂交合成的双二倍体Am6-1和普通小麦品种Ph85-16的回交一代进行产量性状变异特点分析,并利用从130对D基因组SSR引物中筛选出的60对具有多态性的引物对粗山羊草中几个产量性状相关的QTL位点进行了定位,发现粗山羊草的D组染色体对小麦的穗长、千粒重、单株穗数和单株产量具有显著影响,同时初步寻找到6个主效QTL,它们分别为与穗长相关的QSl.sdau-5D,与单株穗数相关的QSnp。sdau-4D、QSnp.sdau-7D,与千粒重相关的QT-gw.sdau-3D,与单株穗数、单株产量相关的Q.sdau-2D-1、Q.sdau-2D-2。     

大豆叶片衰老QTL的初步定位

 【目的】叶片衰老是植物的一个发育性状，对植物体内营养再分配与光合作用有重要影响。【方法】利用2套重组自交系群体进行大豆叶片衰老QTL的初步定位，并进行与产量性状的相关分析。【结果】在C1、D1b+W和O连锁群上检测到3个与大豆叶片衰老有关的QTL，可解释表型变异的5.8%～13.7%。除NJ(SP)BN群体熟期与叶片衰老相关程度较低外，两个RIL群体中生物学产量、籽粒产量、株高和熟期与叶片衰老间均存在显著或极显著的正相关；而收获指数与叶片衰老间一个群体为极显著负相关，另一个群体则表现为极显著正相关。【结论】在大豆育种中，针对延迟叶片衰老选育出的材料有时仅能提高收获指数，这样的材料往往产量潜力不高。