SSR和SNP标记在玉米分子标记辅助背景选择中的应用比较

利用SSR和SNP两种标记对H2321/J076/J076的BC1F1代群体进行分子标记辅助背景选择,比较两种标记在分子标记辅助背景选择上的应用效果。经筛选表明,40对SSR引物中在回交亲本间具有多态性的引物有6对。利用多态性引物对BC1代群体进行毛细管电泳检测结果表明,157个单株轮回亲本背景回复率在50%~100%,背景回复率为100%的单株有1株,背景回复率91.67%的单株有14株。利用Maize SNP3072芯片分析后发现,104个多态性SNP位点可以将157株个体进行精细排序,背景回复率在51.92%~93.75%。比较SSR和SNP标记的背景选择结果,两种标记呈中等程度相关。在回交早代将SSR和SNP标记结合使用进行背景选择,可以实现大群体严选择,提高选择效率。     

分子标记辅助选择下的玉米回交群体农艺性状分析

以综3为受体亲本、衡白522为供体亲本,通过3代回交,利用SSR标记对BC3F1群体背景进行检测,并进行群体植株重要农艺性状的分析。结果表明,群体背景回复率在94.40%以上,平均回复率为97.75%,最高回复率为100%,远远高于经典遗传学背景回复率。农艺性状分析结果显示,背景回复率并未达到分子标记结果。分子标记辅助选择是提高育种选择效果加速育种进程的有效途径之一,针对数量性状控制的农艺性状的选择还需要进一步研究。     

晋麦47背景回交导入系的遗传选择与性状分析

为深入研究小麦抗旱相关性状的遗传基础,以旱地品种晋麦47为轮回亲本、水地品种鲁麦14为供体亲本构建的导入系(ILs)为材料,在雨养和灌溉两种条件下考察穗下节和穗部农艺性状,利用56对在双亲间表现多态性的SSR标记引物对BC3F4导入系群体的150个株系进行遗传选择,分析筛选后的148个导入系遗传背景及性状相关性.结果表明,ILs群体中有两个株系分别在2个、13个标记住点的基因型不同于双亲,ILs群体的晋麦47基因型回复率达94.38%.在两种水分条件下ILs群体多数性状表现超双亲.性状变异系数在2.34%～127.11%之间,性状均值偏向轮回亲本晋麦47;ILs群体灌溉条件下的每穗总小穗数(TNS)极显著大于雨养条件下的,与鲁麦14相似.除穗顶部不育小穗敷(SST)外,其余性状在两种水分条件下均呈极显著正相关(r=0.221～0.555).ILs群体穗下节长(FIL)和旗叶叶枕至德基部长(LPSB)的遗传力均较高,大于0.5975;穗顶部不育小穗数(SST)、穗基部不育小穗数(SSB)的遗传力均较低,小于0.4638,说明小穗结实卒易受环境水分条件影响.     

分子标记辅助选择改良三系杂交稻恢复系R225稻瘟病抗性

稻瘟病是水稻的主要病害,培育抗稻瘟病品种是防治稻瘟病的有效途径。本研究通过分子标记辅助选择与杂交育种相结合的方式,将稻瘟病抗性基因Pi1、Pi2和Pi9导入到三系杂交稻恢复系R225。对BC3F3代材料进行苗期和成熟期稻瘟病抗性鉴定,携带1个或2个抗性基因的目标株系抗性达到中抗以上水平,稻瘟病抗性显著高于各自的轮回亲本。SSR标记分析表明,改良株系的遗传背景回复率达到86.1%~95.3%。通过标记辅助选择获得的改良材料为三系杂交稻恢复系的培育提供了稻瘟病抗性亲本。     

SSR标记辅助回交转育大豆7S球蛋白α-亚基致敏蛋白缺失新品系

为快速培育7S球蛋白致敏蛋白α-亚基缺失的优质大豆新品系,以(α'+α)-亚基双缺失型材料日B为供体亲本,东农47为受体亲本,以杂交-回交转育方法为基础,结合SSR标记辅助遗传背景选择与主要农艺性状鉴定及品质分析,在BC2F4选育到7S球蛋白α-亚基缺失新品系Cb80。Cb80综合农艺性状优良,遗传背景回复率大于90%,其蛋白质及氨基酸总量为44.1%、41.95%,分别比轮回亲本东农47提高3.5和3.76个百分点。     

在构建QPM近等基因系过程中对回交群体的SSR标记选择

优质蛋白玉米(QPM)遗传基础狭窄,以普通玉米自交系为遗传背景培育opaque-2近等基因系是QPM种质改良的重要途径。本文利用SSR标记技术,以opaque-2基因序列为背景开发的特异SSR引物phi057和umc1066作为标记,对构建中的95个QPM近等基因系回交群体BC1F1和BC2F1进行目标基因选择。结果表明,SSR标记phi057对大多数回交群体中的opaque-2基因的选择是有效的;在少数回交群体,如(多黄29×CA335)×多黄29,SSR标记phi057和umc1066不能区分单株间的基因型变异,针对这些材料,需要进行更深入的研究或开发新的SSR。     

基于60K芯片的玉米回交群体精准背景选择策略

利用Maize 6H-60K芯片对京724/京92/京92的BC1代群体进行分子标记辅助背景选择，结合SNP数据特征和回交群体分离特征，开发出一套精准背景选择策略(ABSS:Accurate Background Selection Strategy)及相关算法。从统计结果中选取背景回复程度最高的一批单株作为下一代回交群体的亲本材料。基于Maize 6H-60K芯片分析和筛选产生的28 910个多态性位点，可以对727个样品的背景回复程度进行精细排序，样品背景回复率大部分分布在0.30～0.75(约占样品总数的94.1%)，背景回复率在0.75及以上的样品比例为1.8%。通过分析回交群体重组交换规律以及背景回复率与株高的相关性，进一步验证Maize 6H-60K芯片数据和ABSS的准确性。利用Maize 6H-60K芯片结合ABSS可以精细选择背景回复程度高、重组交换频率相对较低的单株，显著提高背景选择效率，为设计玉米回交群体规模提供参考价值。     

甘蓝型油菜隐性上位互作核不育同型临保系选育新策略

已有研究表明经一定高温处理后,甘蓝型油菜隐性上位互作核不育9012A系统中纯合型不育株稳定彻底,而杂合型不育株发生育性转换,并具有一定的自交结实性。利用这一特性,本研究提出了纯合两用系9012AB的同型临保系选育新策略。以9012AB中不育株为轮回亲本,临保系204TAM为供体亲本进行回交转育。F_1、BC_(1-2)均为不育株群体,从中选择与9012AB相似的不育单株经35℃光照持续处理15h后,用其中育性转换株与9012AB中不育株进行回交转育。在BC3群体中选择与9012AB同型的不育单株,经同样高温胁迫处理后,从育性转换株自交后代的可育株群体中选育出9012AB的同型临保系9012TAM。     

甘蓝型油菜pol CMS育性恢复基因的分子标记

以甘蓝型油菜pol CMS不育系1141A及其恢复系花叶恢为亲本构建F2分离群体，并进一步构建可育和不育分离群体集团。利用RAPD、SSR、AFLP等技术进行标记筛选，获得与Rfp基因连锁的2个分子标记，这2个标记分布于Rfp的两侧，其中，AFLP标记E7P16230与Rfp遗传图距最近，为4.3cM；另一侧的RAPD标记S1-500与Rfp遗传图距为10.8cM。     

高抗稻瘟病品种粤标5号的遗传背景及农艺性状分析

粤标5号是应用分子标记技术育成的抗病优质水稻新品种,应用水稻12条染色体的237对SSR标记对该品种的遗传背景进行分析,结果发现:93个标记在亲本间有明显的多态性,多态性比例为39.2%;在各染色体上,基因型来自轮回亲本的比例为30.0%~100.0%,来自供体亲本的比例为0.0~70.0%;除染色体1和9外,该品种在其它染色体上均与轮回亲本存在不同程度的差异位点,背景回复率为83.3%,低于理论回复率。此外,对粤标5号的农艺性状分析表明,该品种有多个性状介于双亲之间,但在每穗粒数、结实率、千粒重、粒型等与轮回亲本存在极显著差异。研究结果将为水稻抗病分子育种提供参考依据。     

Introgression of Gene for Non-Pollen Type Thermo-Sensitive Genic Male Sterility to Thai Rice Cultivars

For the two-line hybrid rice system, pol en sterility is regulated by recessive gene that responds to temperature. The recessive gene controlling thermo-sensitive genetic male sterility （TGMS） is expressed when the plants are grown in conditions with higher or lower critical temperatures. To transfer tgms gene（s） control ing TGMS to Thai rice cultivars by backcross breeding method, a male sterile line was used as a donor parent while Thai rice cultivars ChaiNat 1, PathumThani 1, and SuphanBuri 1 were used as recurrent parents. The BC2F2 lines were developed from backcrossing and selfing. Moreover, the simple sequence repeat （SSR） markers were developed for identifying tgms gene and the linked marker was used for assisting selection in backcrossing. The identification lines were confirmed by pol en observation. The results showed the success of introgression of the tgms gene into Thai rice cultivars. These lines will be tested for combining ability and used as female parent in hybrid rice production in Thailand.     

玉米矮花叶病抗病基因Rscmv1的精细定位

以感病亲本Mo17为轮回亲本、抗病亲本四一为供体亲本,构建近等基因系,对抗病基因Rscmv1进行精细定位。结果表明,在自交系四一中定位了两个抗病基因Rscmv1和Rscmv2,其中Rscmv1是抗源中普遍存在的主要抗病基因。以选育的近等基因系BC4F3、BC4F4和BC4F5为定位群体,在抗病基因区域内开发了9个有多态性的BAC-SSR标记,将Rscmv1定位在BAC-SSR标记A5-1和B2-1之间,其遗传距离分别为0.3 cM和0.6 cM,2个标记之间的物理距离为1.38 Mb。     

Development of elite breeding lines conferring Bph18 gene-derived resistance to brown planthopper (BPH) by marker-assisted selection and genome-wide background analysis in japonica rice (Oryza sativa L.)

Brown planthopper (BPH) is a serious threat to rice production. In this study, we have used the novel resistance gene Bph18 derived from Oryza australiensis and incorporated it into an elite japonica cultivar, Junambyeo, which is highly susceptible to BPH. The Bph18 gene was introduced by marker-assisted backcross (MAB) breeding into Junambyeo. The backcrossed progenies were evaluated for desirable agronomic and grain quality traits and the selection of improved breeding lines while simultaneously evaluating BPH resistance by bioassays in the greenhouse and foreground selection. Of the 26 advanced backcross breeding lines (ABL), four lines showed agronomic traits similar to those of the recurrent parent, with strong resistance to BPH. Molecular genotyping of the four ABL revealed the conversion of genotypes closely resembling the genotype of Junambyeo. The percentage of donor chromosome segments in ABL decreased from 12.3% in the BC₂ to 9.4%, 8.4% and 5.3% in BC₃, BC₄ and BC₅ generations, respectively. ABL retained small sizes of the donor chromosome segments on chromosomes 1, 2, 10, 11 and 12 but the genomes of ABL2, ABL3 and ABL4 were homosequential to the recurrent parent on chromosomes 3, 4, 5, 6, 7 and 9 without donor chromosome segment introgression. The ABL1 and ABL2 retained only some small segments of the donor genome on chromosomes 9 and 8, respectively. Fine structure analysis of the Bph18 flanking region between RM511 and RM1584 markers on chromosome 12 showed a progressive elimination of donor-derived chromosome segments from BC₂ to BC₅ generations. The percentage of O. australiensis derived chromosome segment substitution in the recurrent parent background decreased from 28% of the donor parent to 6.7%, 3.9%, 3.4% and 3.4% in BC₂, BC₃, BC₄ and BC₅ generations, respectively. However, it was revealed that the O. australiensis-derived chromosome segment (1320 kb) in ABL containing the Bph18 gene was consistently maintained irrespective of advances in backcross generations. BPH resistant elite breeding lines with agronomic and grain quality traits similar to those of the recurrent parent were successfully developed by foreground and background analysis in japonica background without linkage drag.     

栽培稻与普通野生稻BC2F2群体产量相关性状的QTL分析

以广陆矮4号(Oryza sativa ssp. indica)为母本及轮回亲本,普通野生稻（Oryza rufipogon）为父本,分单株连续回交2次,构建BC2F2群体。首先用241对具有双亲多态性的SSR标记对BC2F1单株进行代换片段分析,在此基础上,根据BC2F1的表型选产量较优的单株自交获得BC2F2群体,用代换片段上具有双亲杂合型基因型的24对SSR标记进行QTL定位。在所选的BC2F1单株上,共检测到分布于7条染色体上的20个野生稻的代换片段,平均每条染色体上有2.86个;代换片段长度最小为0.55 cM,最大为33.00 cM,平均长度为12.36 cM,总覆盖长度为247.20 cM,覆盖率为16.21%。利用BC2F2群体对14个产量相关性状进行QTL定位,共检测到控制8个性状的20个QTL。对性状表型值起增效作用的有11个,占总检出数的55%。控制产量相关性状的QTL存在簇状分布现象,这与表型相关分析结果相符合。     

Improvement of Upland Rice Variety by Pyramiding Drought Tolerance QTL with Two Major Blast Resistance Genes for Sustainable Rice Production

Varalu is an early maturing rice variety widely grown in the rainfed ecosystem preferred for its grain type and cooking quality. However, the yield of Varalu is substantially low since it is being affected by reproductive drought stress along with the blast disease. The genetic improvement of Varalu was done by introgressing a major yield QTL, qDTY_12.1, along with two major blast resistance genes i.e. Pi54 and Pi1 through marker-assisted backcross breeding. Both traits were transferred till BC₂ generation and intercrossing was followed to pyramid the two traits. Stringent foreground selection was carried out using linked markers as well as peak markers (RM28099, RM28130, RM511 and RM28163) for the targeted QTL (qDTY_12.1), RM206 for Pi54 and RM224 for Pi1. Extensive background selection was done using genome-wide SSR markers. Six best lines (MSM-36, MSM-49, MSM-53, MSM-57, MSM-60 and MSM-63) having qDTY_12.1 and two blast resistance genes in homozygous condition with recurrent parent genome of 95.0%-96.5% having minimal linkage drag of about 0.1 to 0.7 Mb were identified. These lines showed yield advantage under drought stress as well as irrigated conditions. MSM-36 showed better performance in the national coordinated trials conducted across India, which indicated that improved lines of Varalu expected to replace Varalu and may have an important role in sustaining rice production. The present study demonstrated the successful marker-assisted pyramiding strategy for introgression of genes/QTLs conferring biotic stress resistance and yield under abiotic stress in rice.     

玉米回交后代群体中供体基因组成分分析

以郑58为受体亲本,昌7-2为供体亲本,通过杂交、回交和79对亲本间有多态的SSR标记,分析BC₂F₁和BC₃F₁世代供体的基因组成分。结果表明：BC₂F₁世代单株内的供体染色体片段数平均为11.4个,BC₃F₁世代为4.8个,从BC₂F₁到BC₃F₁单株内的供体染色体片段数减少了57.6%;BC₂F₁世代单株内的供体染色体片段长度平均为99.3 cM,BC₃F₁世代为87.1 cM,BC₃F₁较BC₂F₁减少了12.3%;BC₂F₁世代单株内的供体基因组大小平均为1 132.8 cM,BC3F1世代为421.4 cM,BC₃F₁较BC₂F₁减少了62.8%。