利用染色体片段置换系定位水稻芽期耐冷性QTL

以籼稻品种9311为受体、粳稻品种日本晴为供体构建的95个染色体片段置换系为材料,在5℃低温条件下进行芽期耐冷性鉴定。结果表明,6个置换系低温处理后的成苗率与受体亲本9311有一定差异,其耐冷性略强于9311。利用代换作图法共鉴定出4个与芽期耐冷性相关的QTL,分别位于水稻第5和第7染色体上。其中qCTB-5-1、qCTB-5-2和qCTB-5-3分别被定位在第5染色体RM267与RM1237、RM2422与RM6054及RM3321与RM1054之间遗传距离分别为21.3cM、27.4cM和12.7cM的置换片段上;qCTB-7被定位在第7染色体RM11-RM2752区间遗传距离为6.8cM的置换片段上。     

利用染色体片段置换系定位水稻赖氨酸含量的QTL

 以籼稻品种9311为受体、粳稻品种日本晴为供体构建95个染色体片段置换系, 对水稻控制赖氨酸含量的QTL进行了定位。结果显示, 共有7个染色体片段置换系（chromosome segment substitution line, CSSL）的稻米赖氨酸含量与亲本9311差异显著。利用代换作图法共鉴定了4个与赖氨酸含量相关的QTL, 分别位于水稻第8、9和12染色体上。其中qHLY8和qHLY9.2来自高赖氨酸含量籼稻品种9311, 正向加性效应百分率分别为9.6%和8.5%;而qHLY9.1和qHLY12则来自低赖氨酸含量粳稻品种日本晴, 负向加性效应百分率分别为-16.0%和-21.3%。     

利用染色体片段置换系群体定位和分析水稻粒重和粒型QTL

[目的]挖掘水稻粒重和粒型相关性状QTL,对于解析水稻籽粒遗传机理具有重要作用.[方法]本研究以籼稻9311为受体、粳稻日本晴为供体构建的染色体片段置换系(Chromosome Segment Substitution Lines,CSSLs)群体为材料,在4个环境下对控制稻谷与糙米的粒重和粒型QTL进行了定位分析.[...     

基于重测序的染色体片段代换系定位水稻抽穗期QTL

【目的】水稻的抽穗期是决定水稻产量及其适用性的重要农艺性状之一,是由多基因控制的数量性状。染色体片段代换系减少了个体间遗传背景的干扰,已经成为定位和克隆复杂性状QTL的重要材料。【方法】本研究以9311为受体,日本晴为供体构建的128个重测序的染色体片段代换系群体为试验材料,利用多元回归,结合Bin-map图谱,【结果】鉴定到6个在南京、扬州不同年份间稳定表达的抽穗期QTL,其中,qHD2.1被定位在第2染色体上的759 848 bp区间内;qHD2.2被定位在第2染色体上的45 286 bp区间内;qHD 3.1被定位在第3染色体上的147 931 bp区间内;qHD5.1被定位在第5染色体上的213 351 bp区间内;qHD5.2被定位在第5染色体上的442 305 bp区间内;qHD8.1被定位在第8染色体上的538 176 bp区间内。【结论】本研究为精细定位并克隆相应QTL,进而探明抽穗期QTL的分子调控机制奠定了基础。     

The QTL Analysis of RuBisCO in Flag Leaves and Non-Structural Carbohydrates in Leaf Sheaths of Rice Using Chromosome Segment Substitution Lines and Backcross Progeny F2 Populations

Abstract

In rice (Oryza sativa L.), the maintenance of high photosynthetic rate of flag leaves and the carbon remobilization from leaf sheaths after heading is a critical physiological component affecting the yield. To clarify the genetic basis of RuBisCO content of the flag leaf, a major determinant of photosynthetic rate, and non-structural carbohydrate (NSC) concentration in the third leaf sheath at heading, we carried out quantitative trait loci (QTL) analysis with 39 Koshihikari/Kasalath chromosome segment substitution lines (CSSLs) and backcross progeny F₂ population derived from target CSSL holding the QTL/Koshihikari in the field. QTLs for RuBisCO content and NSC concentration at heading were detected between R2447-C1286 and R2447-R716 on chromosome 10, respectively, by comparing Koshihikari with four CSSLs for chromosome 10 (SL-229, -230, -231 and -232). The progeny QTL for RuBisCO content and for NSC concentration at heading qRCH-10 and qNSCLSH-10-1, respectively, were detected at similar marker intervals between RM8201 and RM5708. In addition, QTLs for RuBisCO content at 14 d after heading, qRCAH-10-1 and qRCAH-10-2, were detected in regions different from that of qRCH-10. No QTL for NSC concentration at 14 d after heading was detected between RM8201 and R716, the region analyzed in this study. The QTLs qRCH-10 and qRCAH-10-1 for RuBisCO content would have additive effects. These QTLs for RuBisCO content and NSC concentration newly found using CSSLs and their backcross progeny F₂ population should be useful for better understanding the genetic basis of source and temporary-sink functions in rice and for genetic improvement of Koshihikari in terms of their functions.     

一个水稻新黄绿叶突变体基因的分子定位

在水稻品种武运粳7号中发现了一个黄绿叶自然突变体,经过多代自交形成了稳定的突变系。该突变系和武运粳7号的正反交F2代的遗传分析表明该材料的黄绿叶由1对隐性基因控制,命名为 ygl 2。利用已有的微卫星（SSR）标记和新发展的SSR标记将 ygl 2基因定位于RM1340、RM7269、RM6298、SSR6 16和RM7434、SSR6 5、SSR6 9、RM5957之间,排列位置为RM1340-RM7269-RM6298-SSR6 16 -ygl 2-RM7434-SSR6 5、SSR6 9-RM5957,它们之间的遗传距离分别为238、0.37、0.00、0.62、0.74、0.49、0.86和1.62 cM,这为 ygl 2基因的分子标记辅助选择育种和图位克隆奠定了基础。     

利用重测序的水稻染色体片段代换系定位控制稻米淀粉黏滞性谱QTL

 稻米RVA谱是评价稻米蒸煮与食用品质的重要指标之一,发掘新的控制稻米RVA谱QTL对稻米品质改良具有重要意义。利用以粳稻品种日本晴为受体、籼稻品种9311为供体并经高通量重测序的染色体片段代换系群体为材料,在两年两点的环境下对该群体中控制RVA谱特征值的QTL进行了定位分析。通过单因素方差分析和Dunnett多重比较, 分析染色体片段代换系与受体亲本之间相关RVA谱特征值的差异,以两年两点都能检测到的显著差异位点作为稳定表达的QTL。共检测到10个稳定表达的QTL,包括控制峰值黏度的4个QTL qPKV2.1、qPKV5.1、qPKV7.1和qPKV8.1,控制热浆黏度的2个QTL qHPV5.1和qHPV7.1,控制冷胶黏度的2个QTL qCPV5.1和qCPV7.1及控制消减值的2个QTL qSBV2.1和qSBV7.1。在两年两点的检测中10个稳定表达QTL的贡献率介于-31.8%～53.2%,这10个QTL分布在第2、5、7和8染色体上,其中位于第2、5和7染色体上的位点存在一因多效性。     

Identification of QTLs for Improvement of Plant Type in Rice (Oryza sativa L.) Using Koshihikari / Kasalath Chromosome Segment Substitution Lines and Backcross Progeny F2 Population

Abstract

Thirty-nine chromosome segment substitution lines (CSSLs) population derived from a Koshihikari / Kasalath cross was used for quantitative trait locus (QTL) analysis of plant type in rice (Oryza sativa L.). Putative rough QTLs (26.2～60.3cM of Kasalath chromosomal segments) for culm length, plant height, panicle number, chlorophyll content of flag leaf blade at heading and specific leaf weight, were mapped on the several chromosomal segments based on the comparison of CSSLs with Koshihikari in the field experiment for 3 years. In order to verify and narrow QTLs detected in CSSLs, we conducted QTL analyses using F₂ populations derived from a cross between Koshihikari and target CSSL holding a putative rough QTL. The qPN-2, QTL for panicle number was mapped on chromosome 2. In traits of flag leaf, the qCHL-4-1 and qCHL-4-2 for chlorophyll content was mapped on chromosome 4, and the qSLW-7 for specific leaf weight on chromosome 7. All QTLs were detected in narrow marker intervals, compared with rough QTLs in CSSLs. The qPN-2, qCHL-4-1 and qCHL-4-2 had only additive effect. On the other hand, the qSLW-7 showed over-dominance. It could be emphasized that QTL analysis in the present study with the combination of CSSLs and backcross progeny F₂ population can not only verify the rough QTLs detected in CSSLs but also estimate allelic effects on the QTL.     

QTL Mapping for Rice RVA Properties Using High-Throughput Re-sequenced Chromosome Segment Substitution Lines

The rapid visco analyser （RVA） profile is an important factor for evaluation of the cooking and eating quality of rice. To improve rice quality, the identification of new quantitative trait loci （QTLs） for RVA profiling is of great significance. We used a japonica rice cultivar Nipponbare as the recipient and indica rice 9311 as the donor to develop a population containing 38 chromosome segment substitution lines （CSSLs） genotyped by a high-throughput re-sequencing strategy. In this study, the population and the parent lines, which contained similar apparent amylose contents, were used to map the QTLs of RVA properties including peak paste viscosity （PKV）, hot paste viscosity （HPV）, cool paste viscosity （CPV）, breakdown viscosity （BKV）, setback viscosity （SBV）, consistency viscosity （CSV）, peak time （PET） and pasting temperature （PAT）. QTL analysis was carried out using one-way analysis of variance and Dunnett＇s test, and stable QTLs were identified over two years and under two environments. We identified 10 stable QTLs： qPKV2-1, qSBV2-1; qPKV5-1, qHPV5-1, qCPV5-1; qPKV7-1, qHPV7-1, qCPV7-1, qSBV7-1; and qPKV8-1 on chromosomes 2, 5, 7 and 8, respectively, with contributions ranging from -95.6% to 47.1%. Besides, there was pleiotropy in the QTLs on chromosomes 2, 5 and 7.     

不同环境下稻米品质性状QTL的检测及稳定性分析

【目的】挖掘新的稻米品质性状QTL并利用分子育种技术改良稻米品质。【方法】利用构建的一套以籼稻香型恢复系昌恢121为背景亲本,以优质粳稻越光为供体亲本的染色体片段代换系（CSSLs）为材料,在4个环境下对稻米品质性状进行QTL检测及稳定性分析。【结果】在4个环境下共检测到44个QTL,其中6个QTL能在多个环境下被检测到;第2、3、5、6和10染色体上存在多效QTL簇,对稻米品质性状具有明显的调控作用;第1、6和12染色体上7个QTL能在不同环境下稳定表达。【结论】第1染色体RM3143-RM1117区间qPGWC1和第12染色体RM3331-RM5479区间qPaT12是两个新的稳定表达QTL。     

不同环境下稻米品质性状QTL的检测及稳定性分析

【目的】挖掘新的稻米品质性状QTL并利用分子育种技术改良稻米品质。【方法】利用构建的一套以籼稻香型恢复系昌恢121为背景亲本，以优质粳稻越光为供体亲本的染色体片段代换系（CSSLs）为材料，在4个环境下对稻米品质性状进行QTL检测及稳定性分析。【结果】在4个环境下共检测到44个QTL，其中6个QTL能在多个环境下被检测到；第2、3、5、6和10染色体上存在多效QTL簇，对稻米品质性状具有明显的调控作用；第1、6和12染色体上7个QTL能在不同环境下稳定表达。【结论】第1染色体RM3143−RM1117区间qPGWC1和第12染色体RM3331−RM5479区间qPaT12是两个新的稳定表达QTL。     

利用重测序的染色体片段代换系群体定位水稻粒型QTL

 以一套用籼稻品种9311为受体、粳稻品种日本晴为供体构建的128个染色体片段代换系为材料,按随机区组实验设计种植,于成熟后考查代换系粒长、粒宽性状,利用多元回归分析方法,结合Bin图谱,鉴定出6个与粒长相关的QTL、2个与粒宽相关的QTL。其中,qGL3.1被定位在水稻第3染色体的5 792 954 bp区间内;qGL3.2被定位在第3染色体的917 878 bp区间内;qGL8.1被定位在第8染色体的889 543 bp区间内;qGL8.2被定位在第8染色体的208 614 bp区间内;qGL9.1被定位在第9染色体的1 149 685 bp区间内;qGL11.1被定位在第11染色体的3 184 760 bp区间内;qGW1.1被定位在第1染色体的200 070 bp区间内;qGW5.1被定位在第5染色体的704 905 bp区间内。上述QTL的准确定位,为进一步精细定位及克隆相应QTL和开展水稻粒型分子育种奠定了基础。     

Assessment of Genetic Diversity of Drought Tolerant and Susceptible Rice Genotypes Using Microsatellite Markers

The introgression of wild chromosomal segments into popular rice varieties is one of the potential approaches for developing varieties for drought stress condition. Sixteen genotypes, including nine indica, two tropical japonica and five chromosome segments substitution lines(CSSLs) with different levels of tolerance/susceptibility to drought stress, were selected for diversity study. Sixty-three microsatellite markers were utilized for assessing genetic diversity. A total of 95 alleles were amplified, and out of them, 60 were polymorphic. Six unique alleles, amplified by the microsatellite loci RM276, RM472,RM488, RM537, RM541 and RM28089, were identified in six genotypes, namely FR13A, Brahamanakhi,RUF44, Swarna-sub1, Brahamanakhi and Satyabhama. The highest genetic similarity was found among CSSLs. Polymorphism information content(PIC) value varied from 0 to 1.00 with an average of 0.66 per locus. Twenty-eight microsatellites were found to be polymorphic, which could be used in marker-assisted selection programme. All the sixteen genotypes were grouped into two major clusters at genetic similarity of 0.64. In the cluster I, five CSSLs identified as diverse genotypes had wild ancestor segments responsible for drought tolerance, and hence they could be utilized as potential donors. The popular Indian varieties, Swarna-sub1 and IR64-sub1, could be used as recurrent parents in the future breeding program for developing varieties for abiotic stresses such as submergence and drought.     

非洲栽培稻垩白粒率耐热性QTL的定位

【目的】本研究旨在定位一个稻米垩白粒率高温耐性QTL,为外观品质育种及解析垩白粒率高温耐性的遗传机制提供依据。【方法】以非洲栽培稻耐热品种IRGC102309(Oryza glaberrima Steud.)和籼稻品种R9311(O. sativa L. subsp. indica Kato.)为亲本构建的栽培稻种间染色体片段导入系CSIL05-23为材料构建次级分离群体,结合人工气候室模拟灌浆期高温胁迫处理,采用垩白粒率高温钝感值为评价指标,对非洲栽培稻垩白粒率高温耐性 QTL 进行检测。【结果】 在BC₆F₂分离群体,利用单标记分析,发现第5染色体上的SSR标记RM1200与垩白粒率耐热性状极显著正相关（P=0.0005）。进一步利用BC₆F₃和BC₆F₄分离群体,采用QTL Cartographer 2.5软件和复合区间作图法在水稻第5染色体上的SSR标记RM1200-RM5796区间重复检测到一个灌浆期垩白粒率耐热性QTL, 命名为qHTCGR5,分别解释11.4%和17.5%表型变异。根据BC₆F₄分离群体的纯合重组体表型分组,利用置换作图方法将目标QTL同样定位在SSR标记RM1200-RM5796之间,遗传图距为1.3 cM,物理图距约为333.4 kb。【结论】 控制垩白粒率耐热性的qHTCGR5是一个能够用于稻米外观品质育种的新QTL。     

利用染色体片段置换系群体检测水稻叶片形态QTL

 水稻叶片的形态改良是水稻株型育种和产量育种的重要目标之一。以9311/日本晴染色体片段置换系（CSSLs）群体为材料,定位了上3叶叶片长、宽、叶面积共9个性状QTL,分析了叶片性状与产量性状之间的相关性,同时定位了主穗重及产量构成因素（颖花数、千粒重、结实率）相关QTL。结果表明,CSSLs群体的叶片性状之间存在显著或极显著相关性;叶片性状与主穗重呈显著或极显著正相关,与主穗颖花数呈极显著正相关;叶片形态多数性状与结实率、千粒重没有显著相关性。两年共定位到20个叶片性状QTL,分布于第1、3、4、5、6、9、11共7条染色体的10个区间,贡献率为3.82%~14.61%,其中贡献率大于10%有6个,多个QTL成簇分布在相同区间,3个QTL在两年间重复检测到,8个QTL为前人未报道的新位点。两年共检测到16个与控制主穗产量相关的QTL,分布于第1、2、3、5、7、8、10共7条染色体13个区间,其中有7个主穗产量相关QTL所在5个区间与叶片形态14个QTL所在区间一致。     

Genetic Analysis and QTL Mapping of Mature Seed Culturability in Indica Rice

Genetic segregation analysis for mature seed culturability was conducted using recombinant inbred lines derived from a cross of indica rice, Yangdao 6 and Pei'ai 64s. Three indices of seed culturability, the frequency of callus induction, the frequency of brown callus and the increase of callus weight were investigated. A combined genetic map constructed with simple sequence repeat (SSR), sequence tag site (STS), cleaved amplified polymorphic sequences (CAPS) and single nucleotide polymorphism (SNP) markers covered a total distance of 1 732.5 cM, averaging approximately 12 cM between two neighboring loci. Three QTLs on chromosomes 7, 7 and 10 were detected for the frequency of callus induction; three QTLs on chromosomes 6, 7 and 9 were detected for the frequency of brown callus; and two QTLs on chromosomes 5 and 7 were detected for the increase of callus weight. Common QTLs mapped at the interval flanking RM5481 and RM6835 on chromosome 7 were identified to be involved in the frequency of callus induction and the frequency of brown callus, explaining 7.29% and 12.52% of phenotypic variation, respectively. A total of 14 epistatic effects were detected for the three indices of mature seed culturability.     

Functional roles of root plasticity and its contribution to water uptake and dry matter production of CSSLs with the genetic background of KDML105 under soil moisture fluctuation

Soil moisture fluctuation (SMF) stress due to erratic rainfall in rainfed lowland (RFL) rice ecosystems negatively affect production. Under such condition, root plasticity is one of the key traits that play important roles for plant adaptation. This study aimed to evaluate root plasticity expression and its functional roles in water uptake, dry matter production and yield under SMF using three chromosome segment substitution lines (CSSLs) with major genetic background of KDML105 and a common substituted segment in chromosome 8. The CSSLs showed greater shoot dry matter production than KDML105 under SMF, which was attributed to the maintenance of stomatal conductance resulting in higher grain yield. The root system development based on total root length of the CSSLs were significantly higher than that of KDML105 due to the promoted production of nodal and lateral roots. These results implied that the common substituted segments in chromosome 8 of the 3 CSSLs may be responsible for the expression of their root plasticity under SMF and contributed to the increase in water uptake and consequently dry matter production and yield. These CSSLs could be used as a good source of genetic material for drought resistance breeding programs targeting rainfed lowland condition with fluctuating soil moisture environments and for further genetic studies to elucidate mechanisms underlying root plasticity.     

Mapping of Rice Fertility-Restoring Genes for ID-Type Cytoplasmic Male Sterility in a Restorer Line R68

An F2 population derived from the cross Zhong 9NR68 was used to map the fertility-restoring (Rf) gene for ID-type cytoplasmic male sterility (CMS).Two bulks (a fertile bulk and a sterile bulk) were constructed by pooling equal amount of ten highly fertile lines and ten highly sterile lines,respectively.Four hundred and thirteen pairs of simple sequence repeat (SSR) primers,which evenly distributed on 12 chromosomes of rice,were selected for analyzing polymorphisms between the parents and between the two bulks.The primer RM283 on chromosome 1 and the primers RM5756,RM258,RM6100 and RM171 on chromosome 10 were found to be polymorphic between the parents and between the two bulks.These five SSR markers were linked to fertility-restoring genes.A total of 82 excessive sterile lines were selected from Zhong 9NR68 F2 population to estimate the genetic distance between five SSR markers and fertility-restoring genes respectively.The results indicated that one Rf gene was linked to RM283 located on chromosome 1 at a distance of 6.7 cM,and the other Rfgene was mapped to the long arm of chromosome 10 flanked by RM258 and RM6100 at the distances of 8.0 cM and 2.4 cM,respectively.     

基于玉米87-1综3单片段代换系的穗长QTL分析

以优良玉米自交系综3为供体、87-1为受体,利用回交和SSR标记辅助选择相结合的方法,获得了51份纯合的染色体单片段代换系(Single Segment Substitution Line,SSSL),每个SSSL内只含有一个来源于供体的染色体片段。51个代换片段不均匀分布在玉米10条染色体上,代换片段长度在1.75～172.45 cM之间,平均长度为24.44cM。覆盖玉米基因组的总长度为933.90 cM,覆盖率为40.20%。2008年在郑州利用17份SSSL材料及在三亚利用39份SSSL材料对玉米穗长进行了1年2点的表型鉴定,共检测到20个穗长QTL,加性效率百分率在-12.10%～19.18%之间。结果表明,这些单片段代换系存在较大的遗传变异,是用于玉米复杂性状的QTL鉴定、基因克隆及功能分析的理想材料。     

Genetic Analysis and Mapping of Dominant Minute Grain Gene Mi3（t） in Rice

Grain size, determined chiefly by grain length, is one of the main factors affecting the grain yield in rice production. To study the trait of rice grain size, F1 and F2 populations were developed from crosses Shuhui 881/Y34 and Shuhui 527/Y34, and genetic analysis for minute grain was performed. The F1 populations showed minute grains, and grain size segregations in the two F2 populations were both in accordance with the ratio of 3：1, indicating that minute grain in Y34 was controlled by a completely dominant gene. By using the F2 population from Shuhui 881/Y34, this dominant gene, tentatively designated as Mi3（t）, was mapped based on SSR markers in the interval between RM282 （genetic distance of 5.1 cM） and RM6283 （genetic distance of 0.9 cM） on the short arm of chromosome 3.