分子标记辅助选择的修饰回交聚合育种方法及其在棉花上的应用

修饰回交育种法是将杂种品系间杂交和回交方法结合起来,用于棉花多个优良性状聚合的育种改良方法。随着分子标记技术日益完善地用于育种的选择中,我们提出了分子标记辅助选择的修饰回交聚合育种方法。它以生产上推广或即将推广的品种为轮回亲本,将修饰回交育种法和分子标记辅助选择育种相结合,同时对轮回亲本的遗传背景和     

The major quantitative trait locus for mungbean yellow mosaic Indian virus resistance is tightly linked in repulsion phase to the major bruchid resistance locus in a cross between mungbean [Vigna radiata (L.) Wilczek] and its wild relative Vigna radiata ssp. sublobata

Mungbean yellow mosaic Indian virus (MYMIV) and bruchid infestation are severe production constraints of mungbean in South Asia, a major global mungbean production area. Marker-assisted selection for resistance against these disorders while maintaining or even improving agronomic traits is an important step toward breeding elite mungbean varieties. This study employed recombinant inbred lines (F12) derived from a cross between MYMIV-tolerant Vigna radiata NM92 and bruchid-resistant V. radiata ssp. sublobata TC1966 to identify chromosomal locations associated with disease and insect pest resistance and seed traits. A linkage map comprising 11 linkage groups was constructed with random amplified polymorphic DNA (RAPD), sequence characterized amplified regions (SCAR), cleaved amplified polymorphic DNA (CAP), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Quantitative trait loci (QTLs) for MYMIV and bruchid resistance, 100 seed weight and seed germination rate were identified. Three major QTLs for MYMIV and one major bruchid resistance locus were mapped on LG 9. The resistance alleles were contributed by the MYMIV tolerant parent NM92 and the bruchid resistant parent TC1966 respectively. One of the MYMIV QTLs was tightly linked in repulsion phase to the bruchid resistance locus. In addition, three minor QTLs for MYMIV resistance were found, where the resistance alleles were contributed by TC1966. Lines combining MYMV resistance alleles from both parents have greater resistance to MYMIV than the tolerant parent. Two minor bruchid resistance QTLs were identified in TC1966. Furthermore, three QTLs each for 100 seed weight and germination rate were detected. The markers defining the QTLs identified in this study will be useful in marker-assisted breeding of improved mungbean varieties in the future.     

烟草六个重要性状的QTL定位

以烤烟台烟7号与白肋烟白肋21作为杂交亲本,后代自交衍生的127个F₂和F_2:3家系为材料,构建了全长3 483 cM包括26个连锁群、190个标记位点的烟草遗传连锁图谱。通过一年两地各3次重复的随机区组田间试验,测定烟叶烟碱、总氯、总钾、叶长、茎叶夹角、白粉病6个重要性状,采用混合线性模型的复合区间作图法定位QTL并分析其遗传互作效应。结果检测到2个烟碱相关QTL、2个总氯相关QTL、1个总钾相关QTL、4个叶长相关QTL、茎叶夹角和白粉病相关QTL各1个,其中6个加性效应QTL和4对加加上位性效应QTL。这11个基因位点遗传效应中除加性效应外,上位性效应也具有重要作用。     

QTL analysis of seed dormancy in rice (Oryza sativa L.)

Effective cumulative temperature (ECT) after heading would be a more reasonable parameter for seed sampling of pre-harvest sprouting/seed dormancy (SD) tests in segregating populations than the days after flowering. SD is an important agronomic trait associated with grain yielding, eating quality and seed quality. To identify genomic regions affecting SD at different grain-filling temperatures, and to further examine the association between SD and ECT during grain-filling, 127 double haploid (DH) lines derived from a cross between ZYQ8 (indica)/JX17 (japonica) by anther culture were analyzed. The quantitative trait loci (QTLs) and their digenic epistasis for SD were identified using a molecular linkage map of this population. A total of four putative QTLs for SD (qSD-3, qSD-5, 6 and 11) were detected on chromosomes 3, 5, 6 and 11, together explaining 41.4% of the phenotypic variation. Nine pairs of digenic epistatic loci were associated with SD on all but chromosome 9, and their contributions to phenotypic variation varied from 2.87%–8.73%. The SD QTL on chromosome 3 was identical to the QTLs found in other mapping populations with different genetic backgrounds, which could be a desirable candidate for gene cloning and marker-assisted selection in rice breeding.     

利用棉花纤维品质相关QTL评价海陆渐渗品种品质初探

 选用第7、13、25号染色体上纤维品质相关QTL(Quantitative trait locus)密集分布区间的48对SSR（Simple sequence repeat）引物,对48份棉花种质进行多态性检测,研究结果显示在实验材料中3对SSR引物具有陆海差异多态性,相关分析表明3个优质基因SSR位点与纤维长度和纤维强度达到极显著相关。通过与海岛棉带型比对、追踪,从分子水平上明确了这些品种中来源于海岛棉渐渗于陆地棉的优异基因区段,为下一步分子标记辅助聚合育种提供了理论参考。     

Comparison and analysis of main effects,epistatic effects,and QTL × environment interactions of QTLs for agronomic traits using DH and RILs populations in rice

Two genetic linkage maps based on doubled haploid (DH) and recombinant inbred lines (RILs) populations, derived from the same indica-japonica cross ‘Samgang × Nagdong’, were constructed to analyze the quantitative trait loci (QTLs) affecting agronomic traits in rice. The segregations of agronomic traits in RILs population showed larger variations than those in DH population. A total of 10 and 12 QTLs were identified on six chromosomes using DH population and seven chromosomes using RILs population, respectively. Three stable QTLs including pl9.1, ph1.1, and gwp11.1 were detected through different years. The percentages of phenotypic variation explained by individual QTLs ranged from 8 to 18% in the DH population and 9 to 33% in the RILs population. Twenty-three epistatic QTLs were identified in the DH population, while 21 epistatic QTLs were detected in the RILs population. Epistatic interactions played an important role in controlling the agronomic traits genetically. Four significant main-effect QTLs were involved in the digenic interactions. Significant interactions between QTLs and environments (QE) were identified in two populations. The QTLs affecting grain weight per panicle (GWP) were more sensitive to the environmental changes. The comparison and QTLs analysis between two populations across different years should help rice breeders to comprehend the genetic mechanisms of quantitative traits and improve breeding programs in marker-assisted selection (MAS).     

Comparison of QTLs mapped in RILs and their test-cross progenies of tropical maize for insect resistance and agronomic traits

Quantitative trait loci (QTL) affecting resistance to south-western corn borer Diatraea grandiosella (SWCB) and sugarcane borer Diatraea saccharalis (SCB) have been identified previously in F_2:3 lines and recombinant inbred lines (RILs) of tropical maize using restriction fragment length polymorphism (RFLP) analyses. Our objective was to determine whether QTLs identified in these generations are also expressed in test crosses (TC) of RILs. A population of 166 TC progenies was developed by crossing RILs from the cross CML131 (susceptible) × CML67 (resistant) with the unrelated, susceptible tester line CML216. Resistance to first-generation SWCB, measured as leaf-feeding damage (LFD) under artificial infestation, and other agronomic traits were evaluated in two environments for the TC progenies and three environments for 183 RILs. The correlation between line per se and TC performance was low for LFD and intermediate for most agronomic traits. Estimates of the genotypic variance and heritabilities were smaller in the TC progenies than in the RILs for all traits. Quantitative trait loci were identified using an RFLP linkage map with 136 loci. For LFD, four QTLs were detected in the TC progenies, of which two were in common with nine QTLs previously mapped in the RILs. Few QTLs for agronomic traits were common to the two types of progeny, because of the low consistency of QTL positions for all traits in RIL and TC progenies, the use of TC progenies should be considered in QTL mapping studies as the first step for marker-assisted selection in hybrid breeding.     

陆地棉产量相关性状的QTL定位

中棉所28和湘杂棉2号分别是以中棉所12×4133和中棉所12×8891配制而成的两个陆地棉强优势杂交种。以其F₂为作图群体,筛选6000多对SSR引物,利用两群体间27个共有多态位点,通过JoinMap 3.0软件整合了一张包含245个多态位点、全长1847.81 cM的遗传图谱。利用Win QTLCart 2.5复合区间作图法分别对两群体8个产量相关性状在F₂和F_2:3中进行QTL定位,在中棉所28群体多环境平均值的联合分析中检测到16个QTL,三环境分离分析中检测到43个QTL;在湘杂棉2号群体分别检测到20个和66个QTL。在A3、D8、D9等染色体上有QTL成簇分布现象,同时在两个群体中发现一些不受环境影响且稳定遗传的QTL。对考察的8个性状在两个群体中发现12对共有QTL,控制果枝数、衣分和籽指的QTL增效基因位点均来源于共同亲本中棉所12。综合分析推测中棉所12的育种价值主要是通过提高后代的结铃性来实现的。研究结果为棉花产量性状的分子设计育种提供了有用的信息。     

Identification of quantitative trait loci for agronomic and physiological traits in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) under high-nitrogen and low-nitrogen conditions

Low-nitrogen (LN) tolerance is a compound character with a complex genetic basis. Many agronomic traits have been shown to be closely related to LN tolerance in maize. In this study, 150 F₇ recombinant inbred lines derived from a cross between inbreds 178 and K12 were evaluated for agronomical and physiological traits under high-nitrogen (HN) and LN conditions in 2 years. Inclusive composite interval mapping (ICIM) was used to identify the quantitative trait loci (QTLs) for traits recorded under different treatments (LN and HN) in 2 years. In total, 86 QTLs were detected: 38 for HN and 35 for LN, while 13 QTLs were detected under both nitrogen levels, suggesting that LN-specific QTLs may play a role in improving LN tolerance in maize. Overlapping QTLs for different traits were located on all chromosomes except chromosome 4 and chromosome 9. Many of these regions overlapped with previously reported QTLs. Several consensus major QTLs and LN-specific major QTLs found in the study can be used in marker-assisted selection breeding for genetic improvement and LN tolerance in maize in the future.     

甜菜抗丛根病育种研究进展

为了研究甜菜抗丛根病育种的抗性基因来源以及作用特点,笔者归纳了甜菜丛根病抗性的来源,包括Alba抗性源、Rizor和Holly抗性源、WB42抗性源、WB41和WB258抗性源以及R36抗性源,并介绍了5种甜菜丛根病的抗性基因特点。另外,对甜菜抗丛根病资源的育成方法进行了总结,包括回交、杂交和轮回选择。通过这些方法将携带抗丛根病资源的甜菜整合至适应性高的甜菜种质资源上,从而获得理想的抗性资源。针对甜菜抗丛根病育种过程中培育和选择甜菜丛根病抗性基因型的过程缓慢、费力且昂贵,以及具有抗性基因品种的抗性易被破坏的情况,现提出如下解决策略：（1）通过分子标记来选择理想的甜菜新品种;（2）不断鉴定新的抗性基因,并将其整合到甜菜的基因库中;（3）开发新抗源,通过鉴定其他基因座上与先前发现的抗性基因的等位基因,为释放具有足够抗性的品种提供机会。抗甜菜丛根病育种的下一个挑战将会是将新的抗性基因导入育种材料,并通过将不同基因座上的多个抗性基因聚合到育种系中进行组合,以最大限度地提高甜菜对丛根病的抗性水平。     

Genetic dissection of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) tiller,plant height,and grain yield based on QTL mapping and metaanalysis

Tiller number per plant (TN) and plant height (PH) are important agronomic traits related to grain yield (GY) in rice (Oryza sativa L.). A total of 30 additive quantitative trait loci (A-QTL) and 9 significant additive × environment interaction QTLs (AE-QTL) were detected, while the phenotypic and QTL correlations confirmed the intrinsic relationship of the three traits. These QTLs were integrated with 986 QTLs from previous studies by metaanalysis. Consensus maps contained 7156 markers for a total map length of 1112.71 cM, onto which 863 QTLs were projected; 78 meta-QTLs (MQTLs) covering 11 of the 30 QTLs were detected from the cross between Dongnong422 and Kongyu131 in this study. A total of 705 predicted genes were distributed over the 21 MQTL intervals with physical length <0.3 Mb; 13 of the 21 MQTLs, and 34 candidate genes related to grain yield and plant development, were screened. Five major QTLs, viz. qGY6-2, qPH7-2, qPH6-3, qTN6-1, and qTN7-1, were not detected in the MQTL intervals and could be used as newly discovered QTLs. Candidate genes within these QTL intervals will play a meaningful role in molecular marker-assisted selection and map-based cloning of rice TN, PH, and GY.     

绿豆分子遗传图谱构建及若干农艺性状的QTL定位分析

利用花叶1号×紫茎1号杂交后代衍生的208个F2家系组建群体,构建含有95个SSR标记位点的遗传连锁图谱,该图谱包含11个连锁群,全长1457.47 c M,标记平均间距为15.34 c M。利用复合区间作图法,对株高、幼茎色、主茎色、生长习性、结荚习性、复叶叶形和成熟叶色等农艺性状进行QTL分析,分别检测到与株高、幼茎色、主茎色、复叶叶形有关的QTL各1个,贡献率在8.49%~66.64%之间;与结荚习性有关的QTL3个,贡献率在60.32%~80.36%之间;与成熟叶色有关QTL 4个,贡献率在69.06%~87.35%之间;与生长习性有关的QTL数量最多,共26个,贡献率在58.32%~99.51%之间。上述QTL主要分布在LG1、LG2、LG4、LG8和LG10连锁群,其中LG1最少,仅检测到生长习性的1个QTL,LG4最多,包含了幼茎色、主茎色、结荚习性、生长习性、复叶叶形、成熟期叶色6个农艺性状的15个QTL;这些QTL既可以应用于绿豆育种的分子标记辅助选择,也对深入研究这些性状的遗传奠定了基础。     

基于全基因组重测序信息开发玉米H99自交系特异分子标记

随着基因组测序技术和计算生物学的发展,利用生物信息学的方法大规模挖掘基因组特异分子标记已成为可能。玉米自交系H99具有较强的可再生能力,是玉米转基因研究的主要供体材料,但是目前对H99基因组信息了解甚少。本研究利用高通量Illumina测序技术,对H99全基因组进行重测序,利用生物信息学手段大规模挖掘并开发了4043个H99特异性的SSR分子标记。随后利用模拟PCR策略,对开发的SSR分子标记进行电子多态性筛选,针对B73×H99、Mo17×H99和B73×Mo17三个群体亲本组合共开发2699候选的特异多态性SSR分子标记。随机挑选20对SSR分子标记对群体亲本B73、H99和Mo17进行多态性筛选,进一步证实候选的多态性具有95%的准确率。此外,基于B73参考序列对开发的多态性SSR分子标记进行全基因组染色体定位及基因注释,揭示了候选多态性SSR分子标记在全基因组及基因内的分布特征。为玉米自交系H99开发了大量的分子标记资源,同时也为快速开发品种间多态性分子标记提供了一套高效可行的分析方法。     

转Cry1Ab基因水稻种质杂交后代的标记辅助选择及抗虫性评价

以转Cry1Mb基因的水稻株系TS．9(后命名为克螟稻)为种质,与优良亲本杂交,在分离世代,通过与目标基因紧密连锁的潮霉素抗性作为筛选标记,结合田间农艺性状选择,获得目标基因位点纯合的新品系。Cry1Mb蛋白含量检测结果表明大多数新品系携带的靶基因在经杂交重组、自交纯合后保留了原亲本高效表达的特性。抗虫鉴定结果表明,对二化螟(Chilo suppressalis)、稻纵卷叶螟等(Cnapha locrocis medinalis)鳞翅目害虫表现高抗性能,同时,在株高、生育期、产量构成因子等重要农艺性状上也有显著改良。从而,对转基因水稻种质的育种利用途径和选择方法作了有益探索。     

QTL mapping and genetic analysis for maize kernel size and weight in multi-environments

Kernel size and weight are important agronomic traits, as well as crucial traits that influence grain yield in maize. In the present study, 150 F₇ recombinant inbred lines derived from a cross 178×K12 were evaluated for kernel length (KL), kernel width (KW), kernel thickness (KT), and 100-kernel weight (HKW) across seven environments. Natural variations in KL, KW, KT, and HKW were observed in the population. A set of quantitative trait loci (QTLs) for the kernel-related traits were identified by inclusive composite interval mapping method. For the four kernel traits from seven environments and the best linear unbiased prediction data, a total of 52 QTLs were detected, which distributed on all chromosomes except chromosome 6. The LOD values ranged from 2.52 to 8.91, the additive effect from ??2.22 to 1.37, and the range of individually explaining phenotypic variation was from 5.8 to 23.49%. Amongst these QTLs, most were detected only in one or two environments. Three stable QTLs, qKL4-1 at bin 4.07/4.08, qKW4-2 at bin 4.06 and qKT2-1 at bin 2.02/2.03, were identified across at least three environments. Besides, several overlapping QTLs associated with multiple traits were identified. For example, qKW3-1 for KW and qHKW3-1 for HKW were located in the same marker interval at Bin 3.01/3.02. These stable QTLs and overlapping QTLs found in this study will contribute to the understanding of genetic components of grain yield and provide the foundation for molecular marker-assisted breeding in maize.     

Herbicide-tolerant crops in agriculture: oilseed rape as a case study

Oilseed rape has been modified extensively by conventional breeding for the production of varieties useful for human consumption (blended vegetable oil and margarine) and industrial processes (rubber additives and high‐temperature lubricants). Because much is now known about its genetic and biochemical composition, it has been an obvious choice for genetic modification and is now at the forefront of the commercial development of genetically modified (GM) or transgenic crops. Around the world, the increase in commercial plantings of all transgenic crops has been rapid. In 1996, 1.7 million hectares were planted, but by 1998 this figure had jumped to 27.8 million ha. The area in the year 2001 is likely to be about 50 million ha. With the possible introduction of transgenic varieties into European agriculture, it is essential that the associated farming practices employed are appropriate for their growth, both from a commercial and an environmental viewpoint. Some of the first transgenic crops are those carrying agronomic traits, e.g. herbicide tolerance transgenes. However, before full commercialization occurs, important agronomic and environmental questions need to be answered. How are these new crops to be incorporated into existing cropping practices? How will this change the current herbicide use profile for a given crop? Do herbicide‐tolerant varieties enhance or impede integrated pest management schemes? What is the likely uptake of such crops in agriculture? What are the ecological implications of their introduction? Are there effective measures to control the spread of transgenes to wild relatives? This paper addresses these questions, with special emphasis on oilseed rape production in the UK, but includes examples from other crops and countries where appropriate.     

利用近缘种属优良基因改良小麦研究进展

小麦是我国以及全球最重要的粮食作物之一。随着人口增长、气候变化和人们对高品质生活的追求,需要培育高产、多抗且优质的小麦品种,确保小麦安全生产和小麦产业健康发展。由于在培育小麦新品种过程中严格持续不断的人工选择,小麦初级基因库已得到充分开发。利用小麦的近缘次级基因库,将传统的育种技术和分子育种技术结合,通过细胞遗传学、分子标记辅助选择和基因工程等方法将小麦近缘种属中的优良基因引入到小麦染色体组中可突破这一育种瓶颈,培育符合当代育种目标的小麦新品种。本文简要介绍了将小麦主要近缘种属及将其蕴含的优良性状和优良外源基因引渗到小麦基因组中的主要技术及特点,重点综述了利用近缘种属优良基因改良小麦抗病性、品质和生长发育等性状的最新研究进展,讨论了目前存在的问题和发展趋势,以期促进小麦近缘种属优良基因的挖掘和新品种培育。     

Breeding tepary bean (Phaseolus acutifolius) for drought adaptation: A review

Tepary bean (Phaseolus acutifolius A. Gray) is a relatively higher drought-tolerant crop than common bean (P. vulgaris), serving as genetic resource for food and genetic enhancement of related legumes. Tepary bean production is hampered by cultivation of low yielding and abiotic stress-susceptible cultivars. Targeted selection of agronomic, physiological and biochemical traits that maximizes yield gains using Phaseolus gene pool is useful to develop stress-tolerant and high-performing genotypes. The objective of this review is to provide breeding progress made regarding tepary bean improvement for drought adaptation. Agronomic, physiological and biochemical traits utilized for selection of drought-tolerant genotypes are highlighted. Genetic and genomic resources developed for tepary bean or closely related species such as common bean useful for genetic analysis and breeding are discussed. Opportunities and challenges to facilitate breeding of tepary bean genotypes with improved abiotic stress adaptation are highlighted. This will enable development of drought-tolerant tepary bean genotypes targeting selection of agronomic, physiological and biochemical traits. Use of genetically related and complementary Phaseolus species and marker-assisted selection method is key to developing drought-tolerant genotypes.     

白肋烟分子标记遗传图谱的构建及部分性状的遗传剖析

用112个AFLP、6个SRAP标记构建了白肋烟的分子标记遗传连锁图谱,包括22个连锁群(A1~A22),总遗传长度为1 953.6 cM,标记的平均间距为20.5 cM,有25个标记表现偏分离(17.0%),主要集中在第1、11和14连锁群上;利用Windows QTL Cartographer Ver. 2.5软件进行QTL分析,结果表明,共检测到11个主效QTL,其中7个与化学成分性状相关,另外4个与农艺性状相关。与烟碱、总氮和总糖相关的QTL各2个(btnic1和btnic2)、2个(bttn1和bttn2)和3个(btts1、btts2和btts3),与株高(btph)、茎围(btls)、节距(btpt)和中部叶长(btl)相关的QTL各1个,没有检测到与总钾、总叶数和中部叶宽相关的QTL,其中与烟碱和总氮相关的QTL处于共分离状态(分别为btnic1和bttn1),表明烟叶烟碱合成与氮素代谢之间可能存在某种未知的生物学相关性,11个主效QTL对各性状表型变异的贡献率在12.3%~26.4%之间。     

Identification of QTLs for agronomic traits using association mapping in lentil

A diverse panel of 96 genotypes of lentil was used in this study to identify QTL for nine agronomic traits through marker-trait association analysis. This study showed significant genetic variability among the lentil genotypes for nine agronomic traits and had medium to large broad sense heritability estimates (h² =?0.58–0.95). Screening of 534 SSR markers resulted in 266 polymorphic loci that generated 697 alleles ranging from 2 to 16 alleles per locus across the genotypes. The model-based population structure analysis identified two distinct subpopulations among lentil genotypes and each subpopulation did not show any admixture. Marker trait association (MTA) analysis following ML model resulted in the identification of 24 MTAs for nine traits at P?<?0.01. The per cent of phenotypic variation explained by each associated marker with particular agronomic trait ranged from 7.3 to 25.8%. The highest proportion of total phenotypic variation (23.1–25.8%) was explained by the QTLs controlling the primary branches/per plant. In the present study, few EST-SSR markers showed significant association with days to maturity, pods/plant, secondary branches/plant, 100 seed weight, yield/plant and reproductive duration and explained large phenotypic variation (7.3–23.8%). Hence, these markers can be used as functional markers in lentil breeding program for developing improved cultivars.