大豆异黄酮及其组分含量的遗传分析与QTL检测

以栽培大豆晋豆23为母本,以山西农家品种大豆灰布支黑豆为父本杂交衍生的447个RIL作为供试群体构建遗传图谱,利用高效液相色谱法定性、定量测定样品中的异黄酮及其组分含量。采用主基因+多基因混合遗传分离分析法和Win QTLCart 2.5复合区间作图法,对大豆异黄酮及其组分含量进行混合遗传分析和QTL定位。结果表明,大豆苷、黄豆苷元、染料木素、染料木苷、大豆苷元和异黄酮总含量分别受4、4、2、3、2和2对主基因控制,并有多基因修饰。检测到44个与大豆异黄酮及其组分含量相关的QTL,与大豆苷、染料木素、黄豆苷元、大豆苷元、染料木苷和异黄酮总含量相关的QTL分别有10、9、4、7、8和6个。连续2年分别检测到与大豆苷、染料木苷、黄豆苷元和异黄酮关联,分别位于标记区间satt430~satt359、satt038~satt570、satt197~sat_128和satt249~satt285的稳定表达QTL,可尝试用于分子标记辅助育种。     

大豆品种成熟期基因型推测的研究

选择不同来源中国大豆品种23份和国外引进的成熟期近等基因系35份进行SSR分析,目的是鉴定与成熟期基因型紧密连锁的标记,进而推测中国大豆品种的成熟期基因型。结果表明,（1）210对SSR标记中125对在成熟期近等基因型中具有多态性,推测与成熟期有关的标记有8个;（2）在Clark近等基因系中,筛选出成熟期基因E3/e3特异性标记Satt229,E4/e4的特异SSR标记Sct_010、Satt294、Satt247、Satt452和Satt156;在Clark和Harosoy近等基因系中,筛选出E7/e7的特异性SSR标记为Satt071、Satt178;（3）根据8个与成熟期相关的标记的分子数据,构建了国外大豆近等基因系的UPGMA聚类图,共聚为4类,背景来源相同或相似的材料被聚为一类,明显分为Clark近等基因系和Harosoy近等基因系。（4）与近等基因系成熟期基因(E7)分子标记比对,推测出25份中国大豆品种的成熟期基因。     

大豆脂肪酸主要组分含量QTL定位

以中黄13×中黄20的100个BC₂F₂家系为作图群体,构建了一张包含131个SSR分子标记的遗传连锁图谱,图谱总长为2157.3 cM,平均遗传距离为16.5 cM,涵盖了大豆的20个连锁群。利用气相色谱技术测定BC₂F₂、BC₂F₃和BC₂F₄回交群体的脂肪酸主要组分含量,采用IciMapping 3.3完备区间作图法定位QTL,共检测到5种脂肪酸组分相关的QTL 26个,与棕榈酸、硬脂酸、油酸、亚油酸和亚麻酸相关的QTL分别为5、5、7、5和4个;3个区间在不同年份被检测到与同一脂肪酸组分相关,sat_294~satt228连续3年被检测到与棕榈酸含量相关,sat_253~satt323和sat_292~satt397连续2年被检测到与油酸含量相关;4个区间被检测到与2种脂肪酸组分相关,其中sat_294~satt228与棕榈酸、油酸相关,satt308~sat_422与硬脂酸、亚油酸相关,sat_292~satt397与油酸、亚油酸相关,satt374~satt269与油酸、亚麻酸相关。     

控制水稻品种Koshihikari抽穗期的数量性状位点

利用Koshihikari × Kasalath BILs群体及相应的Kasalath/Koshihikari CSSLs群体,在江苏南京和海南陵水两个环境下对抽穗期的QTL定位分析。结果表明,在两地重复检测到3个QTL,分别位于第3、6和8染色体上;在qHd-3和qHd-8位点,来自Koshihikari等位基因能够提早抽穗,在qHd-6位点,来自Koshihikari等位基因能够延迟抽穗;第3染色体qHd-3和第7染色体非加性QTL之间存在上位性互作,通过重组自交系和置换系相互验证发现,qHd-3 所在的标记区间与W008的Kasalath插入片段位置大体一致,qHd-8所在的标记区间与W023、W024的Kasalath插入片段相吻合,qHd-7-1所在的标记区间位于W20的Kasalath片段之内,表明确实存在这3个位点。同时还对Koshihikari × 桂朝2号RILs抽穗期的QTL定位分析,在南京和海南分别检测到3个加性抽穗期QTL,1对非加性抽穗期QTL存在互作。     

云南高原粳稻抗白叶枯病新品系云资抗21号的选育

以高产优质的云南高原粳稻品种滇系4号为轮回亲本，以IRBB21为白叶枯病抗性基因Xa21的供体亲本，通过杂交和3次回交以及5次自交形成BC3F，代材料，并结合后代材料的白叶枯病抗性鉴定及农艺性状的评价选择，选育出高原粳稻新品系云资抗21号，对云资抗21号的21个单株采用与Xa21连锁标记RM21和RM229、以及22个独立分离的SSR标记进行检测，结果表明这21个单株在RM21位点上均为IRBB21的纯合基因型，在RM229位点上有1个单株显示为轮回亲本的纯合基因型，其余的20个单株均为IRBB21的纯合基因型，而在其余22个SSR标记位点上均显示为轮回亲本的纯合基因型。该新品系白叶枯病抗性与IRBB21相近，农艺性状与轮回亲本相似，具有很好的应用前景。     

采用SSR标记辅助选育具有Xa22（t）的云南高原粳稻新种质

以高原粳稻新品种滇粳优1号为轮回亲本，携带Xa22（t）基因的云南地方稻种扎昌龙为供体亲本，回交后代BC3F1290个株行和BC3F2290个株行为供试材料，采用与Xa22（t）紧密连锁标记RM224及其它1l对SSR标记进行辅助选择。290株BC3F1个体在RM224位点上杂合基因型个体的比率为21．04％；其它11对SSR标记位点的轮回亲本纯合基因型个体比率平均为93．07％，但不同染色体标记位点上纯合基因型的比率不同。用云南高原粳稻上的白叶枯病优势菌株YH24对亲本、BC3F1单株和BC3F2株行接种鉴定，     

大白菜抗根肿病近等基因系的选育及其评价

本研究以具有抗根肿病基因CRb的大白菜‘CR Shinki DH’系为供体亲本,大白菜自交系‘91-12B’为轮回亲本,通过回交和4个阶段的分子标记辅助选择,选育出7个携带CRb基因的抗根肿病近等基因系.通过CRb基因所在大白菜A3染色体的分子标记检测和抗病性鉴定,筛选出抗根肿病、并插入有供体亲本最短染色体片段的3个近等基因系B1S1、B2S2和B3S1.表型分析表明这3个近等基因系的结球相关性状与轮回亲本无显著差异,而且保持了‘91-12B’的配合力.所选育的近等基因系不仅能应用于育种实践,也为通过转录组分析研究根肿病的抗病机理提供了材料.     

小麦抗白粉病基因Pm43定位区段内RGA分析

利用普通小麦测序草图可从基因组范围内对小麦单条染色体上的某个区段进行分析。Pm43是作物遗传与分子改良山西省重点实验室在小麦2D染色体长臂上定位的一个抗白粉病基因。利用信息学方法分析Pm43所在物理图谱、遗传图谱和基因组图谱上的位置,可为其精细定位乃至候选基因的确定提供参考。试验采用Pm43两侧标记序列进行比对,将Pm43定位于染色体C-2DL3-0.49区间的79~99 c M内,所在基因组区段为2DL_9835990~2DL_9823315。利用目前已克隆小麦抗病基因的保守基序作为探针,从目标区段内检索出89条包含抗病基因类似物(Resistance gene analogues,RGA)序列的scaffold,其中,36条scaffold被诊断出含有SSR位点,之后针对SSR位点开发分子标记。利用携带有Pm43的普通小麦材料CH5025、感白粉病材料台长29以及CH5025×台长29的F2作图群体的抗感池DNA,对开发的SSR标记进行连锁性检测,共筛选出4个多态性标记,从而将目标区段进一步确定在标记PK_9908430和NBS_9908778之间。最后经聚类分析,筛选出与已克隆Pm基因同源性较高的1个PK序列和1个NBS序列,且在粗山羊草2D染色体和水稻第4染色体中均存在与这2个序列同源的RGA表达序列。     

基于SLAF-seq技术开发长穗偃麦草染色体特异分子标记

长穗偃麦草1E及7E染色体上带有重要的抗赤霉病基因, 开发大量相关染色体特异分子标记有助于准确定位抗性基因及获得可用于辅助育种紧密连锁的标记。基于SLAF-seq技术, 获得了368个长穗偃麦草1E染色体特异片段, 随机选取80个特异片段设计引物, 开发了20个长穗偃麦草1E染色体特异分子标记、2个长穗偃麦草基因组特异分子标记及26个其他特异分子标记, 效率达60%。用这些特异标记能稳定检测出不同小麦–长穗偃麦草衍生材料中的1E染色体或片段。通过标记与优良性状的共分离特性, 获得与相关基因紧密连锁的标记, 将为小麦抗性育种中的分子标记辅助选择提供依据。     

水稻野败型细胞质雄性不育恢复基因Rf3的定位

以珍汕97A/明恢63的F2群体为材料,应用SSR标记对水稻野败型恢复基因Rf3进行定位。该试验从F2分离群体中筛选出119个极端不育单株组成隐性基因定位群体。针对水稻第1染色体短臂Rf3所在染色体的可能区间,应用37个SSR标记检测亲本,从16个多态性标记中挑选出9个检测定位群体。结果表明物理位置连续排列的SSR标记RM10353、RM1195和RM3746各有8个单株与Rf3基因发生了单交换,且重组子数表现为最少,据此可将Rf3定位于这3个标记的两侧标记内。因此最终将Rf3定位在相距679.9 kb的SSR标记RM10338和RM10376之间。     

不同光周期条件下大豆生育期主基因的效应

以大豆生育期近等基因系为材料, 比较12 h短日照(SD)及16 h长日照(LD)条件下E1/e1、E2/e2、E3/e3、E4/e4、E5/e5、E7/e7等6对生育期相关主基因的效应。结果表明, 在大多数生育期基因型中, 显性位点延迟大豆的开花期和成熟期, 隐性位点提早开花期和成熟期。同一基因在不同遗传背景下的效应值不同, 显性位点可增强其他基因的效应, 说明各基因间存在互作。生育期基因的效应受光周期影响很大, 长日照可增强大豆生育期相关基因的效应, 短日照则相反。此外, 光周期对基因效应的影响因发育阶段不同而变化, 其中, E1基因在大豆营养生长阶段、E4基因在生殖生长阶段受光周期影响较大, 而E3基因在营养生长和生殖发育阶段均受光周期的严格调控。不同光照条件下生育期基因效应的分析结果, 可为不同生态区大豆品种生育期性状的定量设计提供依据。     

Feasibility of selection for high weed suppressive ability in soybean: Absence of tradeoffs between rapid initial growth and sustained later growth

In ecology, tradeoff theory has been used to understand differences among plant species in their competitive abilities. In efforts to develop weed-suppressive soybean, we found evidence of a tradeoff between the ability for rapid initial growth and the ability for sustained growth later in the season: early maturing lines displayed more rapid initial growth but ceased growth sooner. Such a tradeoff would increase the difficulty of obtaining a full-season weed-suppressive variety. To determine this tradeoff's existence and severity we examined two possible mechanisms that could lead to it. We tested whether early maturing soybean lines attain higher early relative growth rate than late-maturing soybean lines and whether early maturing soybean lines produce larger seeds by an environmentally-dependent or -independent mechanism. Early maturing lines had higher relative stem elongation rates than late lines but not higher relative dry weight or leaf area increase rates. In more northern locations and in years with shorter growing seasons, early maturity lines produced larger seeds than late maturing lines, implicating an environmentally-dependent rather than -independent mechanism causing seed size differences. Relative to early lines, when late lines mature, temperature and photoperiod are in greater decline, leading to a risk of incomplete seed fill. Resulting seed size decreases could lead to lower initial growth in late maturity lines and thus to an environmental rather than genetic or physiological cause of tradeoff between initial growth and sustained growth later in the season. This revised version was published online in July 2006 with corrections to the Cover Date.     

转几丁质酶和核糖体失活蛋白双价基因大豆的获得与抗病性鉴定

以东北地区4个主栽大豆品种为试材,通过农杆菌介导法,将2个抗真菌病基因,即几丁质酶基因和核糖体失活蛋白基因构建在同一植物表达载体上,对受体材料进行遗传转化,获得了经Southern检测同时整合双价基因的T₀代转基因大豆植株,转化频率为0.5%。将T₀代转基因大豆植株扩繁至T₂代,并进行了大豆疫霉根腐病和大豆灰斑病的抗性检测,4个株系的抗性均比对照有明显提高。分子生物学验证及RT-PCR检测表明2个外源基因在4个株系中均进行了转录表达。     

Genetic and molecular bases of photoperiod responses of flowering in soybean

Flowering is one of the most important processes involved in crop adaptation and productivity. A number of major genes and quantitative trait loci (QTLs) for flowering have been reported in soybean (Glycine max). These genes and QTLs interact with one another and with the environment to greatly influence not only flowering and maturity but also plant morphology, final yield, and stress tolerance. The information available on the soybean genome sequence and on the molecular bases of flowering in Arabidopsis will undoubtedly facilitate the molecular dissection of flowering in soybean. Here, we review the present status of our understanding of the genetic and molecular mechanisms of flowering in soybean. We also discuss our identification of orthologs of Arabidopsis flowering genes from among the 46,367 genes annotated in the publicly available soybean genome database Phytozome Glyma 1.0. We emphasize the usefulness of a combined approach including QTL analysis, fine mapping, and use of candidate gene information from model plant species in genetic and molecular studies of soybean flowering.     

不同熟期及产量类型的大豆生殖生长期生理特性的比较研究

通过分析不同熟期、不同产量类型的大豆，特别是高产品种（系）的群体生理特性，为栽培措施的合理运用及生态育种指标的有效选择提供理论参考。连续2年的试验结果表明，高产大豆应具有株高适宜，节数相对较多，产量形成的空间较大，生殖生长期LAI和LAD较高，R5期后叶片衰减慢，叶片在各个方向上的分布均匀，干物质积累量大，     

反义RNA介导GmFAD2-1B基因沉默增强大豆种子中油酸的高效累积

油酸含量是评价大豆油食用品质和稳定性的重要指标。本研究采用农杆菌介导转化法,将反义GmFAD2-1B基因导入栽培大豆品种,获得油酸含量显著提高的转基因大豆新品系。Southern杂交检测表明,外源GmFAD2-1B基因片段已导入大豆基因组,其插入拷贝数为1~5个。q RT-PCR检测表明,外源GmFAD2-1B主要在大豆种子中表达,并导致种子中内源GmFAD2-1 m RNA表达水平显著降低,而根、茎、叶、花组织中内源GmFAD2-1 m RNA表达水平无显著变化。脂肪酸组分分析表明,12份转基因大豆种子油酸含量为27.38%~80.42%,其中,L40和L72油酸含量分别为68.91%~80.42%和65.98%~80.22%,较对照品种Williams 82(17.8%~22.0%)提高2.65倍以上,亚油酸降低至4.84%~14.55%,饱和脂肪酸降低至10.34%~11.16%,但总脂肪和总蛋白含量与对照品种相比没有显著变化。农艺性状分析表明,转基因大豆在熟期、株高、叶形、花色、结荚高度、百粒重等方面与对照品种也没有显著差异。     

Identification of soybean genes related to fatty acid content based on a soybean genome collinearity analysis

Seed fatty acid content is an important consideration for soybean produced for food, feed, and industrial applications. In this study, MCScanX was used to analyze the entire soybean genome to generate a collinearity block, which was then used to assess the collinearity among the soybean fatty acid quantitative trait loci (QTL) in the SoyBase database. The hub‐QTLs located in the Gm06, Gm07, and Gm10 segments were identified. The Kyoto Encyclopedia of Genes and Genomes and gene ontology databases were used to analyze the genes in hub‐QTL regions, resulting in the identification of 17 candidate genes related to soybean fatty acid content. Two lines with different fatty acid contents and a recurrent parent were selected from a chromosome segment substitution line library for a subsequent quantitative real‐time polymerase chain reaction (qRT‐PCR) assay to verify the candidate gene expression patterns. Four genes were related to the total soybean fatty acid content, while three genes were related to the content of specific fatty acid types. The results of this study may be relevant for the fine mapping of soybean fatty acid QTLs/genes.     

Effect of maturity and temperature on breeding for mid-oleic soybean lines under the high heat conditions of the Mississippi Delta,USA

Soybean (Glycine max (L) Merr.) seed is an important source of oil for human consumption. Increasing the percentage of oleic acid in soybean seed oil is an important breeding objective because increasing the oleic acid content improves the oxidative stability of the oil. Extensive literature shows that temperature during seed-fill is positively correlated with the content of oleic acid in soybeans. In addition, it was shown that a maturity QTL was linked to an oleic acid QTL. The Mississippi Delta in the USA is a hot environment where soybean harvest begins in August, which is the hottest part of the season. The purpose of this research was to determine the possibility of developing both early- and late-maturing lines with consistent >?50% oleic acid content in Mississippi. We selected early and late segregants from three genetically different breeding populations also segregating for mid-oleic acid derived from crosses to germplasm N98-4445A, a non-transgenic freely available line with >?50% oleic acid. The selected lines were grown in 2 years in three trials at Stoneville, MS. Results indicated that no late-maturing lines (MG V) met the targeted mid-oleic acid level, whereas MG III and early MG IV lines with oleic acid over 50% were obtained. No maturity-alone effect on oleic acid content was observed, due to the bias of the strong negative correlation between maturity date and mean temperature during seed-fill. This study demonstrated that breeders can effectively develop early soybeans with oleic acid levels greater than 50% for the midsouthern USA.     

Multivariate analysis of a collection of soybean cultivars for southwestern Spain

Summary Multivariate techniques were used to classify 125 soybean lines into clusters. Late maturing varieties belonging to maturity group III showed the best adaptation to the ecological conditions of the area when soybeans were sown at early planting dates. A group of experimental lines, the majority of which had semideterminate stem termination with small leaflets and intermediate maturity, were highly productive when grown as a second crop. Some lines of other groups were identified as likely parents for use in a breeding program to improve agronomic characteristics. The identified groups were quite stable in their performance through changes in environmental conditions (years and planting dates). The analyses indicated that some intercorrelated traits can be omitted in future line evaluation.Experimental work and data analyses supported by the Instituto Nacional de Investigaciones Agrarias of Spain and the IRI Research Institute, Inc. through a World Bank Grant.INIA, Moncada (Valencia), Spain INIA, Madrid, Spain; and Iowa State University, Ames, Iowa 50011. Consultant with IRI Research Institute, Inc. (1975–77).     

Implications of the population effect in the selection of soybean progeny

The goal of this study was to verify the influence of the population effect in the estimates of genetic and phenotypic components and to identify the best soybean progeny or lines in a commercial soybean breeding programme. We evaluated 292 populations for grain yield and absolute maturity during three agricultural years. To quantify the efficiency of the inclusion of the population's effect in the model, we estimated genetic gain with the selection, the Spearman's correlation, the coincidence index, the realized gain and correlated response of selected genotypes with and without the effect of the population. It was found that the variance components, heritability and coefficient of experimental variation were better estimated when the effect of the population was included, providing greater gain with selection for grain yield and absolute maturity. Coincidence and ranking among the selected progeny with and without the effect of the population are of greater magnitude in more advanced inbreeding generations and at higher percentage of selected progeny. The use of the population effect has greater importance in earlier generations of inbreeding.