Analysis of rice blast resistance genes by QTL mapping

Resistance to rice blast pathogen mostly shows a quantitative trait controlled by several minor genes. Its complexity and the mutable characteristic of rice blast isolates both hinder the development of the blast resistance research. The article here tried to explore the resistance gene distribution on rice chromosomes and the way of function. Totally 124 QTLs have been identified against 20 isolates using Cartographer software with a ZYQ8/JX17 DH population, which separately are at 100 loci of 72 marker intervals on 12 rice chromosomes. Of them, 16 QTLs were determined by the isolate HB-97-36-1. 82 QTLs (66.13%) are from the resistant parent alleles, ZYQ8, while 42 QTLs (33.87%) are from the susceptible parent alleles, JX17. In comparison of their positions on chromosome, most QTLs are clustered together and distributed nearby the major genes especially the regions on chromosomes 1, 2, 8, 10 and 12. Each QTL could account for the resistance variation between 3~2%-68.64%. And, a positional QTL might display the resistance to several different isolates with different contributions.     

Isolation and identification of the three rice monotelosomics

In order to obtain rice monotelosomic, the progeny of 24 telotrisomics, derived from an indica rice variety, Zhongxian 3037, were screened. The variants that differed morphologically from the diploids and the original primary trisomics as well as the telotrisomics were collected for cytological identification. The variants with 24 chromosomes were selected according to the prometaphase chromosomes. From these variants, three monotelosomies with one chromosome arm deletion in each were verified by fluorescence in situ hybridization （FISH） using a rice centromeric BAC clone of 17p22 as a marker probe. The three monotelosomics were derived from telotrisomic 1S, 4L and 11L, respectively. Further identification was conducted on the prometaphase or pachytene chromosomes of the three variants, which were probed with the same centromeric BAC clone together with the corresponding chromosome arm specific makers, a0059H02 （on the short arm of chromosome 1）, a0034E24 （on the long arm of chromosome 4）, and a0071H11 （on the long arm of chromosome 11）. The results indicated that the telocentric chromosomes in the three monotelosom. ics were derived from their respective corresponding telotrisomics. According to the telocentric chromosomes of the variants, they were monotelosomic 1S （one long arm of chromosome 1 was lost）, monotelosomic 4L （one short arm of chromosome 4 was lost） and monotelosomic 11L （one short arm of chromosome 11 was lost）, respectively.     

Identification and mapping of quantitative trait loci controlling cold-tolerance of Chinese common with rice （O.rufipogon Griff.） at booting to flowering stages

An advanced backcross population of rice was used to identify the quantitative trait locus(QTL) controlling the cold-tolerance at booting to flowering stages.The recipient,Guichao 2(GC2),was a commercial Indica rice;the donor Dongxiang common wild rice,was an accession of common wild rice(DXCWR,Oryza rufipogon Griff.).Three QTLs for cold-tolerance were detected on chromosomes 1,6 and 11.Two of them coming from DXCWR could enhance the cold-tolerance of the backcross progenies.Moreover,one sterility QTL that could reduce the seed set rate of the backcross progenies by 78% was mapped on chromosome5.     

QTL analysis of flag leaf in barley （Hordeum vulgate L.） for morphological traits and chlorophyll content

To understand genetic patterns of the morphological and physiological traits in flag leaf of barley, a double haploid （DH） population derived from the parents Yerong and Franklin was used to determine quantitative trait loci （QTL） controlling length, width, length/width, and chlorophyll content of flag leaves. A total of 9 QTLs showing significantly additive effect were detected in 8 intervals on 5 chromosomes. The variation of individual QTL ranged from 1.9% to 20.2%. For chlorophyll content expressed as SPAD value, 4 QTLs were identified on chromosomes 2H, 3H and 6H; for leaf length and width, 2 QTLs located on chromosomes 5H and 7H, and 2 QTLs located on chromosome 5H were detected; and for length/width, I QTL was detected on chromosome 7H. The identification of these QTLs associated with the properties of flag leaf is useful for barley improvement in breeding programs.     

Quantitative trait loci （QTLs） for quality traits related to protein and starch in wheat

Quality traits in wheat （Triticum aestirum L.） were studied by quantitative trait locus （QTL） analysis in a recombinant inbred line （RIL） population, a set of 131 lines derived from Chuan 35050 × Shannong 483 cross （ChSh）. Grains from RILs were assayed for 21 quality traits related to protein and starch. A total of 35 putative QTLs for 19 traits with a single QTL explaining 7.99-40.52% of phenotypic variations were detected on 10 chromosomes, 1D, 2A, 2D, 3B, 3D, 5A, 6A, 6B, 6D, and 7B. The additive effects of 30 QTLs were positive, contributed by Chuan 35050, the remaining 5 QTLs were negative with the additive effect contributed by Shannong 483. For protein traits, 15 QTLs were obtained and most of them were located on chromosomes 1 D, 3B and 6D, while 20 QTLs for starch traits were detected and most of them were located on chromosomes 3D, 6B and 7B. Only 7 QTLs for protein and starch traits were co-located in three regions on chromosomes 1D, 2A and 2D. These protein and starch trait QTLs showed a distinct distribution pattern in certain regions and chromosomes. Twenty-two QTLs were clustered in 6 regions of 5 chromosomes. Two QTL clusters for protein traits were located on chromosomes 1D and 3B, respectively, three clusters for starch traits on chromosomes 3D, 6B and 7B, and one cluster including protein and starch traits on chromosome 1D.     

Comparative mapping of rice root traits in seedlings grown in nutrient or non-nutrient solution

The component and amount of nutrient in the growth medium are the major factors affecting root growth.For the systematic dissection of root gene expression,evaluation of nutrient and non-nutrient solutions was conducted for their effect on root traits and quantitative trait loci(QTL)mapping.Three rice root parameters,maximum root length(MRL),root dry weight(RDW),and root/ shoot ratio of dry weight(RSR),were characterized within a double haploid(DH)population from a cross of ZYQ8(indica)and JX17(japonica).The value of the three root traits in two parents all decreased under the nutrient condition compared to those under the nonnutrient condition,of which RSR decreased up to 2.6-fold on average.In the DH population,more than 70 % lines in MRL,94 % lines in RDW,and all the lines in RSR were scored lower.In total,eight QTLs were identified in nutrient system(5 from JX17 alleles and 3 from ZYQ8 alleles)while five QTLs were detected in non-nutrient system(4 from JX17 alleles and 1 from ZYQ8 alleles).Of them,one QTL for RSR was shared by both culturing systems,seven QTLs were specific in nutrient system and the other four QTLs were specific in non-nutrient system.All 13 QTLs were distributed over 7 rice chromosomes-2,3,4,5,6,9 and 10,respectively.     

Comparative mapping of QTLs for Al tolerance in rice and identification of positional Al-induced genes

Aluminum (A1) toxicity is the major factor limiting crop productivity in acid soils. In this study, a recombinant inbreed line (RIL) population derived from a cross between an A1 sensitive lowland indica rice variety IR1552 and an A1 tolerant upland japonica rice variety Azucena, was used for mapping quantitative trait loci (QTLs) for A1 tolerance. Three QTLs for relative root length (RRL) were detected on chromosome 1,9, 12, respectively, and 1 QTL for root length under A1 stress is identical on chromosome 1 after one week and two weeks stress. Comparison of QTLs on chromosome 1 from different studies indicated an identical interval between C86 and RZ801 with gene(s) for A1 tolerance. This interval provides an important start point for isolating genes responsible for A1 tolerance and understanding the genetic nature of Al tolerance in rice. Four A1 induced ESTs located in this interval were screened by reverse Northern analysis and confirmed by Northern analysis. They would be candidate genes for the QTL.     

Generating of rice OsCENH3-GFP transgenic plants and their genetic applications

In order to investigate rice functional centromeres, OsCENH3-GFP chimeric gene was constructed and transformed into the indica rice variety, Zhongxian 3037, mediated by Agrobacturium. The integration of the exogenous genes in the transgenic plants was confirmed by PCR and Southern blotting. The transgenic plants grow normally during their whole life time, just like Zhongxian 3037. No significant defects were detected in either mitosis or meiosis of the transgenic plants. The overlapping of GFP signals and anti-CENH3 foci in both mitotic and meiotic cells from T0 and T1 generation plants indicated that GFP had been successfully fused with CENH3, so the GFP signals can well represent the CENH3 locations on each chromosome. To evaluate the applicability of the transgenic plants to other genetic studies, fluorescence in situ hybridization （FISH） using rice centromeric tandem repetitive sequence CentO as the probe was conducted on the zygotene chromosomes of pollen mother cells （PMCs）. It has been revealed that the GFP signals are overlapping with CentO FISH signals, showing that CentO is one of the key elements constituting rice functional centromeres. Immunofluorescent staining using anti-o-tublin antibody and anti-PAIR2 antibody on the chromosomes during mitosis and meiosis stages of the transgenic plants further reveals that OsCENH3-GFP transgenic plants can be widely used for studying rice molecular biology, especially for tagging functional centromeres in both living cells and tissues.     

Generating of rice OsCENH3-GFP transgenic plants and their genetic applications

In order to investigate rice functional centromeres, OsCENH3-GFP chimeric gene was constructed and transformed into the indica rice variety, Zhongxian 3037, mediated by Agrobacturium. The integration of the exogenous genes in the transgenic plants was confirmed by PCR and Southern blotting. The transgenic plants grow normally during their whole life time, just like Zhongxian 3037. No significant defects were detected in either mitosis or meiosis of the transgenic plants. The overlapping of GFP signals and anti-CENH3 foci in both mitotic and meiotic cells from T0 and T1 generation plants indicated that GFP had been successfully fused with CENH3, so the GFP signals can well represent the CENH3 locations on each chromosome. To evaluate the applicability of the transgenic plants to other genetic studies, fluorescence in situ hybridization (FISH) using rice centromeric tandem repetitive sequence CentO as the probe was conducted on the zygotene chromosomes of pollen mother cells (PMCs). It has been revealed that the GFP signals are overlapping with CentO FISH signals, showing that CentO is one of the key elements constituting rice functional centromeres. Immunofluorescent staining using anti-α-tublin antibody and anti-PAIR2 antibody on the chromosomes during mitosis and meiosis stages of the transgenic plants further reveals that OsCENH3-GFP transgenic plants can be widely used for studying rice molecular biology, especially for tagging functional centromeres in both living cells and tissues.     

Construction of a genetic map and localization of QTLs for yield traits in tomato by SSR markers

Using an F2 population derived from the hybrid of Lycopersicon esculentum Mill. ‘XF 98-7’× Lycopersicon pimpinellifolium LA2184, a SSR genetic linkage map of tomato is constructed. The map contains 112 markers and spans 808.4 cM with an average distance of 7.22 cM between loci. Two quantitative trait loci (QTLs) for first flower node on chromosomes 5 and 11, two QTLs for number of flowers per truss on chromosomes 2 and 5, and five QTLs for fruit weight on chromosomes 1, 2, 3, 9 and 12 are identified.     

Tagging major quantitative trait loci for sheath blight resistance in a rice variety, Jasmine 85

This study was conducted with a clonal F₂ population of rice from a cross between Jasmine 85, a resistant variety, and Lemont, a susceptible cultivar. The rice plants belonging to each F₂ clone were divided into two plots, which were put in two replicates, respectively. Clonal parents were tested as controls. The plants were inoculated by short toothpicks incubated with RH-9, a virulent isolate of the pathogenic fungus,Rhizoctonia solani, which causes rice sheath blight. The extreme resistant and susceptible clonal lines were selected for construction of resistant and susceptible DNA pools, respectively. A total of 94 polymorphic markers evenly distributed on 12 rice chromosomes were used for bulked segregant analysis, three positive ones were found polymorphic between the two DNA pools, and three major QTLs for sheath blight resistance, Rh-2, Rh-3 and Rh-7, were identified. The three major QTLs were located on chromosomes 2, 3 and 7, and could explain 14.4%, 26.1% and 22.2% of the phenotypic variation.     

水稻芒长及其分布特征相关QTL的定位

水稻芒的性状与驯化过程密切相关,具体表现为芒长短和芒的分布等．利用一个无芒品种越光（轮回亲本）和一个有芒品种Kasalath杂交产生的回交重组自交系群体,在上海、海南两地对芒长和芒的分布（芒出现比例）进行了相关分析及其相关基因（QTL）定位,结果发现这两个性状间存在显著的正相关．共检测到影响这两个性状的12个QTL,分布在6条染色体上的7个区域,贡献率介于2．90％-53．24％,其正效应大多来自有芒亲本Kasalath,但也有3个QTL正效应来自无芒亲本越光,并对这个现象产生的原因进行了讨论．本研究检测到的QTL及其两侧的分子标记可以用于水稻驯化的研究和理想型无芒品种分子辅助育种．     

玉米穗部性状的QTL定位

以玉米自交系L26和095组配的Fz世代为定位群体,采用SSR分子标记技术构建了包括98个位点的连锁图谱,结合F2穗部性状的鉴定结果,利用复合区间作图法对秃尖长等8个穗部性状进行基因定位,共检出21个QTL.其中穗长检测到3个QTL;穗粗、穗行数分别检测到2个QTL;行粒数检测到3个QTL;轴粗检测到2个QTL;200粒质量检测到3个QTL;穗粒质量检测到6个QTL;秃尖长没有检测到QTL.检出的21个QTL中,有10个QTL的解释变异率超过了20%,表现为主效QTL效应.研究还发现,穗部性状QTL在玉米10条染色体上分布不均匀,且成簇分布.该试验中检测到的21个QTL中,有10个影响不同性状的QTL位于3个染色体区域.各个QTL位点上起增、减效作用的等位基因在亲本间分布不均匀.     

控制水稻剑叶形态相关性状的数量基因位点(QTL)的定位

水稻籽粒中一半以上的碳水化合物来自剑叶的光合作用，剑叶形态改良一直是水稻株型育种的一个重要目标．利用一个日本主要种植的粳稻品种越光（轮回亲本）和一个印度的籼稻品种Kasalath杂交产生的回交重组自交系群体（backcross recombinant inbred lines，BILs）对剑叶形态中的3个主要性状（剑叶长、叶宽以及其叶面积）进行了相关分析及其数量基因位点（quantitative trait loci，QTL）的定位．研究表明，控制剑叶形态的3个主要性状间存在极显著的正相关，并检测到影响3个性状的8个QTL，分布在第1，3，4，6条染色体上，贡献率介于4．94％～22．07％，其中第4染色体上C1016标记和第6染色体上C556标记附近的共有6个QTL，其两侧的紧密分子标记在水稻株型分子育种上具有一定应用价值．     

水稻苗期形态性状的QTL定位

采用苗期形态性状上有着明显差异的粳稻Asominori和籼稻IR24杂交产生的重组自交系（RILs）为材料,经营养液培养后,对水稻苗期形态相关的6个数量性状：最大根长（MRL）、苗高（SH）、根干重（RDW）、茎（叶）干重（SDW）、总重（TW）以及根/茎（叶）干重比（RSR）进行了数量性状位点（QTL）定位分析.结果共检测到影响6个形态性状的8个QTL位点,分别位于水稻的第2,4,10和11染色体上,其贡献率为9.79%～22.90%.并发现位于第4染色体上的控制根/茎（叶）干重比的qRSR-4和根干重的qRDW-4的2个位点以及位于第10染色体上的控制茎（叶）干重的qSDW-10和总重的qTW-10的2个位点分别位于同一分子标记区间.研究结果对水稻苗期形态性状QTL的精细定位及培育理想株型具有一定的应用价值.